je fl Pras ysievacth “Pye Med ce eee Site en an feapedeters #4 fa hat O21. MACE, eta PROCEEDINGS of the LINNEAN SOCIETY NEW SOUTH WALES VOLUME 119 March 1998 JUN 25 1998 Proc. LINN. SOc. N.S.W., 119. 1998 The Relationship of the Australian Freshwater Crayfish Genera Euastacus and Astacopsis SUSAN H. LAWLER! AND KEITH A. CRANDALL? (Communicated by J.R. Merrick) 'Department of Environmental Management and Ecology, La Trobe University, PO Box 821, Wodonga, VIC 3689; and *Department of Zoology and Monte L. Bean Museum, Brigham Young University, Provo, Utah 84602 USA Lawler, S.H. and Crandall, K.A. (1998). The relationship of the Australian freshwater cray- fish genera Euastacus and Astacopsis. Proceedings of the Linnean Society of New South Wales 119, 1-8. The genera Euastacus and Astacopsis are thought to be closely related because of their similar morphologies and ecologies as well as their location in southeastern Australia. Members of the genus Astacopsis are restricted to Tasmania whereas Euastacus ranges from northern Cape York to southern Victoria. In order to test for the monophyly of each genus and to examine the evolutionary relationships among genera, DNA sequences from the 16S region of the mitochondrial rDNA array from members of these two genera were compared. Our data indicate that the genera are evolutionarily distinct. Astacopsis appears to be para- phyletic, with members of the genus Euastacus forming a monophyletic group within the Astacopsis. Manuscript received 21 August 1997, accepted for publication 5 January 1998. Keywords: Euastacus, Astacopsis, freshwater crayfish, phylogeny, mitochondrial DNA, maximum likelihood. INTRODUCTION The genera Euastacus and Astacopsis are both called spiny crayfish and are known to prefer cool, pristine freshwater habitats. The genus Euastacus 1s widespread in eastern mainland Australia, while Astacopsis 1s restricted to Tasmania. Both genera have been recently revised. The genus Astacopsis is now thought to contain three species (Hamr 1992), although in the past as few as two were recognised (Swain et al. 1982) and as many as four (Riek 1969). The genus Euastacus is much larger and variable with 41 species currently recognised (Morgan 1986, 1988, 1989, 1997). According to Riek (1972), the two genera are sister taxa and their closest relative is the genus Astacoides, which is restricted to Madagascar. The spiny crayfishes have much in common both morphologically and ecologically. Species distributions closely correspond to river drainages, with high endemism throughout southeast Australia (Merrick 1993, 1995). Clark (1936) divided the two genera based on the telson (membranous in Euastacus and calcareous in Astacopsis), the stems of podobranchs (wing-like in Euvastacus) and the relative spininess of the abdomen. Nevertheless, the monophyly of the genus Astacopsis has been questioned (Horwitz 1996), reflecting a long-standing concern that the two genera do not form natural species groups. Many of the morphological characters used in taxonomy and phylogeny of these species are highly variable or subject to convergent evolution. Attempts to divide species of Astacopsis based on spininess found that this character was influenced by both the habitat and geographic region in which the animals were collected (Swain et al. 1982). Proc. LINN. SOc. N.S.W., 119. 1998 i) EUSTACUS AND ASTACOPSIS Since the ecological requirements of these genera are virtually identical, they may have similar morphologies for reasons other than taxonomic relationship. Convergent evolution in morphological characters is known to occur in many crayfish groups (Hobbs 1974). Australian freshwater crayfish can be broadly separated into true burrowers, which hold their chelae in a vertical plane, and the moderate burrowers, whose chelae are hori- zontal. The moderate burrowers include the genera Euastacus, Astacopsis, Euastacoides, Astacoides, Cherax and Paranephrops (Riek 1972). Of these, Astacoides is presumed to be the most closely related (Riek 1972), but it is only found in Madagascar and tissue was not available. Paranephrops occurs only in New Zealand, and samples were unavail- able for analysis. Thus the genus chosen as an outgroup was Cherax, primarily because it is phylogenetically distinct from Euastacus and Astacopsis and because multiple species were available, allowing a comparison of genetic diversity among genera. Others have attempted to identify natural phylogenetic groups within the freshwa- ter crayfish using molecular characters. Patak and Baldwin (1984) examined the relation- ships of 6 freshwater crayfish genera using distances generated using antibody/antigen reactions and electrophoretic domains of the blood protein haemocyanin. The genera Euastacus and Astacopsis could not be distinguished using this approach, although the genus Cherax was found to be genetically distinct and basal to the other two genera. However, using data from 30 allozyme loci from 7 species of Euastacus and all 3 species of Astacopsis, Avery and Austin (1997) found more differences between the genera than between species within each genus, supporting their current taxonomic status. This study, based on DNA sequences from the 16S mitochondrial region, tests whether Astacopsis is distinct from Euastacus and provides a preliminary look at rela- tionships among species within these two genera using the genus Cherax as an outgroup. TABLE 1 Species used and the location from which they were collected. Species Location Astacopsis franklinii New Town Rivulet, Northern Hobart, Tasmania A. tricornis Huon River, Western Tasmania Cherax cuspidatus Bell Creek Rd., Caloundra, Queensland C. destructor albidus Barney Creek south of Halls Gap, Victoria C. robustus Crayhaven Yabbie Farm, North Arm Cove, NSW Euastacus armatus Hoy River, Harrietville, Victoria E. australasiensis Wirreanda Creek, north of Church Point, NSW E. bispinosus Burrong Falls off of Rose Creek Rd., Victoria E. yarraensis Upper Gellibrand River, west of Barramunga, Victoria MATERIALS AND METHODS Crayfish of the genera Euastacus, Astacopsis and Cherax were collected from locations throughout eastern Australia, usually by turning rocks and catching individuals by hand (see Table 1). The genus Cherax was included in the study as an outgroup because this genus is clearly phylogenetically distinct from Euastacus and Astacopsis, and because Cherax has been hypothesized to be most closely related to Euastacus (Riek 1969; Crandall et al. 1995; Patak and Baldwin 1984). Proc. LINN. SOC. N.S.W., 119. 1998 S.H. LAWLER AND K.A. CRANDALL 3 Tissue from the gills and tail were frozen or preserved in ethanol. DNA was extracted and the 16S region of the mitochondria was amplified via the polymerase chain reaction using standard protocols (Crandall et al. 1995). PCR reaction conditions consisted of an ini- tial two minute denaturation at 92°C, followed by 30 cycles of one minute denature at 92°C, 30 second annealing at 45°C, and 30 second extension at 72°C. These 30 cycles were then followed by a 10 minute extension at 72°C. PCR products were sequenced from both ends with an ABI 377 automated sequencer, following the manufacturer’s instructions. Finally, the sequences were edited and spliced together by eye to make a single contiguous ~520 base pair unit. Sequences were aligned using Clustal W (Thompson et al. 1994). Unrooted phylogenies, with Cherax specified as the outgroup, were estimated using the maximum parsimony approach with equal weights assigned to all changes (PAUP*, 4.0d56: Swofford 1997). Weighting schemes that incorporated the observed transition bias had no effect on the tree topology. An exhaustive search was performed which examines every possible tree topology (PAUP*, 4.0d56: Swofford 1997). Phylogenetic relationships were also estimated using maximum likelihood (Felsenstein, 1981) and neighbor-joining (Saitou and Nei 1987) as implemented by PAUP* (Swofford 1997). The model of evolution used in these analyses was determined by the procedure outlined in Huelsenbeck and Crandall (1997); namely, a likelihood ratio test was used to determine significant differences among models of evolution. The likelihood ratio statis- tic, | = —2(InLg — InL}), was compared to a ¥~ distribution with a Bonferroni adjusted significance level for multiple comparisons. Phylogenetic signal was measured via the gy statistic (Hillis and Huelsenbeck 1992). Relative amounts of genetic diversity in the different genus lineages was measured using the approach of Crozier (1992), as implemented by the computer program Conserve 3.0 (Agapow 1997). Branch lengths were estimated as the proportion of overall diversity within the phylogeny (Crozier and Kusmierski 1994). Confidence in resulting clades was assessed using the bootstrap procedure (Felsenstein 1985). The testing of alternative phylogenetic hypotheses was performed using a sign test (Crandall and Fitzpatrick 1996) which is an unweighted version of the Wilcoxon signed rank test (Templeton 1983). RESULTS The resulting sequences have been deposited in Genbank under accession numbers AF044240-AF044248. The exhaustive parsimony search resulted in a single most parsi- monious tree (Fig. 1). The gy statistic showed significant skewness in the tree distribu- tion (—0.8575) indicating significant phylogenetic signal (P < 0.01, Hillis and Huelsenbeck 1992). This phylogenetic signal remained even after constraining clades with high bootstrap support. Several molecular evolutionary hypotheses were tested using these sequence data, as shown in Table 2. Firstly we rejected the hypothesis of equal base frequencies as our data show an A/T bias (A = 0.322, T = 0.353, C = 0.108, and G = 0.217). Likewise, equal rates of transitions and transversions were rejected, with these sequences showing a transition/transversion ratio of 2.023 (Table 2). We were also able to reject the hypothe- sis of equal evolutionary rates among sites, since incorporating a model with the gamma distribution increased the likelihood over the null hypothesis of equal rates among sites (G shape parameter = 0.384, Table 2). Incorporating the proportion of invariant sites into the model does not significantly increase our likelihood. Thus we concluded that the most appropriate model of evolution for our data was the HK Y85+G model (for details, see Swofford et al. 1996). Using this model of evolution we estimated the maximum likelihood tree to be identical to the parsimony tree (Fig. 1). The same tree was also esti- mated using the neighbor-joining method. Proc. LINN. Soc. N.S.W., 119. 1998 4 EUSTACUS AND ASTACOPSIS TABLE 2 Test of molecular hypotheses to determine the appropriate model of evolution for maximum likelihood and neighbor-joining searches. The Bonferroni adjusted significance level for five comparisons is a = 0.01. Null Hypothesis Models Compared -InLO -InL1 -2IndN sf. P Equal base frequencies Ho: JC69 1957.46 1904.51 105.9 3. 4.14x 10-23 Hy: F81 Equal transition/transversion rates Hg: F81 1904.51 1863.10 82.82 1 4.55 x 10-20 H,: HKY85 Equal rates among sites Ho: HKY85 1863.10 1826.31 73.58 1 4.90x 10-18 Hy: HKY85+T Proportion of invariable sites Ho: HKY85+T 1826.31 1826.31 0 1 1 Hy: HK Y85+4I +invar Molecular Clock Ho: HKY85+T'c 1836.73 1826.31 20.84 7 1.57 x 10-3 H,: HKY85+P The tree presented in Figure | indicates that the genus Astacopsis is paraphyletic with respect to Euastacus. Euastacus appears to be monophyletic, although the maxi- mum likelihood bootstrap support was only 61% for this grouping. Significance of bootstrapping values is open to interpretation (Hillis and Bull 1993), so tests of these results were done using a sign test to compare several alternative phylogenetic hypothe- ses of Astacopsis and Euastacus monophyly and nonmonophyly (Fig. 2). The hypothe- sis that both Astacopsis and Euastacus are monophyletic can be rejected at P = 0.0592 (Fig. 2A). The hypothesis that both Ewastacus and Astacopsis are both paraphyletic can be rejected at the P = 0.0898 level of significance (Fig. 2B). Finally, we can test the hypothesis that Astacopsis is monophyletic and derived from the paraphyletic Euastacus (Fig. 2C). We reject this hypothesis at the P = 0.0119 level of significance. Clearly these tests are only marginally significant. Future work will incorporate more taxa and more sequence data, including data from nuclear genes, to further test these hypotheses. Thus our sequence data supports the monophyly of Euastacus and the para- phyly of Astacopsis. Genetic diversity was measured for each lineage. The lowest genetic diversity was found in the genus Euastacus (GD = 0.0924) despite having the largest number of species represented. Astacopsis was intermediate in genetic diversity (GD = 0.168), almost dou- bling the value for Euastacus. Finally, Cherax had the highest diversity (0.2269). The molecular clock test rejected the hypothesis of equal rates among all lineages, however this rejection was not highly significant (Table 2). Differences in rates of evolution would not alter the fact that the high genetic diversity in Astacopsis should elevate its conserva- tion status, and the imperilment of A. gouldi support this point (Horwitz 1994). DISCUSSION Unfortunately our efforts to amplify DNA from A. gouldi were not successful, so all three species of Astacopsis are not represented in our phylogeny. Nevertheless, the data presented here suggest that the genus is evolutionarily distinct from Euastacus. If Proc. LINN. SOc. N.S.W., 119. 1998 S.H. LAWLER AND K.A. CRANDALL Nn Astacopsis tricornis Euastacus bispinosus Euastacus yarraensis Euastacus armatus Euastacus australasie Astacopsis franklini Cherax destructor albidus Cherax rotundus Cherax cuspidatus j= = 5 nucleotide substitutions Figure 1. The maximum parsimony, maximum likelihood and neighborhood joining tree is shown with branch lengths proportional to the amount of nucleotide divergence along each branch. The scale at the bottom of the figure gives an indication of the number of substitutions occurring along each branch. The tree is unrooted with the Cherax species designated as the outgroup. The relative support for each clade is shown as a bootstrap per- centage at the nodes with the parsimony bootstrap values shown on top and the maximum likelihood bootstrap values shown in parentheses. The bootstrap values were based on 1000 replications. we root our phylogeny with the Euastacus clade (data not shown), then the Astacopsis are not even in the same clade as Euastacus and appear ancestral to Cherax. Our phy- logeny based on two Astacopsis and four Euastacus species indicates that the mainland genus Euastacus is a monophyletic group within the Astacopsis, which is paraphyletic. The highest genetic diversity was found in the genus Cherax, but the fact that Astacopsis has almost twice the genetic diversity of Euastacus supports the idea that Astacopsis is an older evolutionary lineage from which Euastacus may be recently derived. This result is especially surprising because the two Astacopsis species sequenced are difficult to distinguish morphologically, and were previously thought to be very closely related, even conspecific (Swain et al. 1982). Our data give the genus Astacopsis a high conservation priority from a genetic perspective (Crozier 1992). Avery and Austin (1997) were unable to distinguish between A. tricornis and A. franklinii within the genus Astacopsis, and E. bispinosus, E. armatus and E. yarraensis within Euastacus using allozyme electrophoresis. Although our mitochondrial data sup- Proc. LINN. Soc. N.S.W., 119. 1998 6 EUSTACUS AND ASTACOPSIS Eustacus bispinosus E. yarraensis E. armatus E. australasiensis Astacpsis tricornis A. franklinii Charax destructor albidus C. rotundus A C. cuspidatus Astacpsis tricornis Eustacus australasiensis E. bispinosus E. yarraensis E. armatus A. franklinii Charax destructor albidus C. rotundus C. cuspidatus Eustacus australasiensis E. armatus E. bispinosus Astacpsis tricornis A. franklinii E. yarraensis Charax destructor albidus Cc C. rotundus C. cuspidatus Figure 2. The alternative phylogenetic hypotheses tested. A) Astacopsis and Euastacus are monophyletic, B) Astacopsis and Euastacus are paraphyletic (this phylogeny differs from Figure 1 in the placement of E. aus- tralasiensis between the two Astacopsis, making Euastacus paraphyletic), and C) Astacopsis is monophyletic and derived from the paraphyletic Euastacus. Proc. LINN. Soc. N.S.W., 119. 1998 S.H. LAWLER AND K.A. CRANDALL 7 port the close relationship between the three species of Euastacus (notice the short branch lengths in Fig. 1), the two species of Astacopsis are genetically quite distinct. The primary vicariance event for Tasmanian invertebrates seems to be the high country of central Tasmania, which would have been periglacial as recently as 7000 years ago (Mesibov 1994). Thus the major split among faunal components in Tasmania is east/west, reflecting the distribution of the species Astacopsis tricornis and A. franklinii (Hamr 1992). Our data showing that these two species are distinct support this vicariance event for the genus Astacopsis. Astacopsis gouldi, which occurs along the northern coast of Tasmania, nevertheless does not occur in the Tamar river system in northcentral Tasmania, and therefore has distinct western and eastern metapopulations (Horwitz 1994). Sequence data from these populations would be of considerable interest. Hypotheses of relationships among members of Astacopsis and Euastacus have focused on Euastacus species in southeastern Victoria. The last link between Tasmania and Victoria about 12,000 years ago across Flinders Island to Wilson’s Promontory (Williams 1974:171), would have allowed contact between the Tasmanian and southeast Victorian freshwater crayfish. Four species of Euastacus from southeastern Victoria (E. bidawalus, E. diversus, E. neodiversus and E. woiwuru) share a morphological character (the male cuticle partition) with the Astacopsis (Morgan 1983). Thus there may be a sub- set of Euastacus species that are more like Astacopsis, and sequence data from these would be of particular interest. Preliminary data (not shown) place E. bidawalus firmly within the Euastacus cluster, supporting the two genera as they stand. Horwitz (1996) suggested that A. franklinii is more closely related to Euastacus woiwuru and E. neodiversus than to A. gouldi and A. tricornis, based on morphological characters in a dichotomous key. Testing of these hypotheses will require sequence data from more taxa. In summary, this study indicates that the genus Astacopsis 1s not monophyletic, nor is it a derived branch of the genus Euastacus. Further research will include sequencing A. gouldi and more species of Euastacus, particularly the species that have a male cuticle partition and come from southeastern Victoria. ACKNOWLEDGEMENTS We thank Margie Kinnersley for providing some of the sequence data. Alistair Richardson provided tissues of Astacopsis gouldi and Chris Austin provided tissues of A. franklinii. We also thank Drs Richardson and Austin, Pierre Horwitz and Barb Mable for assistance in collecting the specimens used in this study. This work was sup- ported by the Alfred P. Sloan Foundation, the National Science Foundation, and Deakin University (KAC). REFERENCES Agapow, P. (1997). Conserve. 3.0.1. LaTrobe University, Melbourne, Australia. Avery, L. and Austin, C. M. (1997). The biochemical taxonomy of the spiny crayfish genera Astacopsis and Euastacus in south eastern Australia. Memoirs of the Museum of Victoria 56, 543-555. Clark, E. (1936). The freshwater and land crayfishes of Australia. Memoirs of the National Museum of Victoria 10, 5-56. Crandall, K.A., Lawler, S.H. and Austin, C.M. (1995). A preliminary examination of the molecular phylogenet- ic relationships of some crayfish genera from Australia (Decapoda: Parastacidae). Freshwater Crayfish 10, 18-30. Crandall, K.A. and Fitzpatrick, J.F., Jr. (1996). Crayfish molecular systematics: Using a combination of proce- dures to estimate phylogeny. Systematic Biology 45, 1-26. Crozier, R.H. (1992). Genetic diversity and the agony of choice. Biological Conservation 61, 11-15. Crozier, R.H. and Kusmierski, R.M. (1994). Genetic distances and the setting of conservation priorities. In Conservation Genetics (Eds V. Loeschcke, J. Tomiuk and S.K. Jain). pp. 227—237. Birkhauser Verlag, Basel. Felsenstein, J. (1981). Evolutionary trees from DNA sequences: A maximum likelihood approach. Journal of Molecular Evolution 17, 368-376. Proc. LINN. SOc. N.S.W., 119. 1998 8 EUSTACUS AND ASTACOPSIS Felsenstein, J. (1985). Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39, 783-791. Hamr, P. (1992). A revision of the Tasmanian freshwater crayfish genus Astacopsis Huxley (Decapoda: Parastacidae). Papers and Proceedings of the Royal Society of Tasmania 126, 91-94. Hillis, D.M. and Bull, J.J. (1993). An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. Systematic Biology 42, 182-192. Hillis, D.M. and Huelsenbeck, J.P. (1992). Signal, noise, and reliability in molecular phylogenetic analyses. Journal of Heredity 83, 189-195. Hobbs, H.H. Jr. (1974). Adaptations and convergence in North American crayfishes. Freshwater Crayfish 2, 541-551. Horwitz, P. (1988). An assessment of the Caroline Creek freshwater crayfish reserve in northern Tasmania. Papers and Proceedings of the Royal Society of Tasmania. 122, 69-72. Horwitz, P. (1994). Distribution and conservation status of the Tasmanian giant freshwater lobster Astacopsis gouldi (Decapoda: Parastacidae). Biological Conservation 69, 199-206. Horwitz, P. (1996). Biogeographical affinities of macrocrusteacean groups in northeast Tasmania. Records of the Queen Victoria Museum and Art Gallery, No. 103. Huelsenbeck, J.P. and Crandall, K.A. (1997). Phylogeny estimation and hypothesis testing using maximum likelihood. Annual Review of Ecology and Systematics 28, in press. Merrick, J.R. (1993). Freshwater crayfishes of New South Wales. Linnean Society of NSW, Milson’s Point, NSW. 127 pp. Merrick, J.R. (1995). Diversity, distribution and conservation of freshwater crayfishes in the eastern highlands of New South Wales. Proceedings of the Linnean Society of NSW 115, 247-258. Mesibov, R. (1994). Faunal breaks in Tasmania and their significance for invertebrate conservation. Memoirs of the Queensland Museum 36, 133-136. Morgan, G.J. (1983). A taxonomic revision of the freshwater crayfish genus Euastacus Clark (Decapoda, Parastacidae) . PhD thesis, Monash University, Melbourne. Morgan, G.J. (1986). Freshwater crayfish of the genus Euastacus Clark (Decapoda, Parastacidae) from Victoria. Memoirs of the Museum of Victoria 47, 1-57. Morgan, G.J. (1988). 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The phylogeny of the Parastacidae (Crustacea: Astacoidea), and descriptions of a new genus of Australian freshwater crayfishes. Australian Journal of Zoology 20, 369-389. Saitou, N. and Nei, M. (1987). The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution 4, 406-425. Swain, R., Richardson, A.M.M. and Hortle, M. (1982). Revision of the Tasmanian genus of freshwater crayfish Astacopsis Huxley (Decapoda: Parastacidae). Australian Journal of Marine and Freshwater Research 33, 699-709. Swofford, D.L. (1997). PAUP* Phylogenetic analysis using parsimony and other methods. 4.0.0d56. Sinauer Associates, Sunderland, MA. Swofford, D.L., Olsen, G.J., Waddell, P.J. and Hillis, D.M. (1996). Phylogenetic Inference. In Molecular Systematics (Eds D.M. Hillis, C. Moritz and B.K. Mable). pp. 407-514. Sinauer Associates, Inc., Sunderland, MA. Templeton, A.R. (1983). Phylogenetic inference from restriction endonuclease cleavage site maps with particu- lar reference to the evolution of humans and the apes. Evolution 37, 221-244. Thompson, J.D., Higgins, D.G. and Gibson, T.J. (1994). CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research 22, 4673-4680. Williams, W.D. (1974). Biogeography and Ecology in Tasmania. Dr. W. Junk, b.v., The Hague, Netherlands. 498 pp. Proc. LINN. SOC. N.S.W., 119. 1998 Evolution of Queensland Spiny Mountain Crayfish of the Genus Euastacus Clark (Decapoda: Parastacidae): Preliminary 16s mtDNA Phylogeny MARK PONNIAH AND JANE M. HUGHES (Communicated by J.R. Merrick) Cooperative Research Centre for Tropical Rainforest Ecology and Management, Faculty of Environmental Sciences, Griffith University, Nathan QLD 4111 PONNIAH, M. AND HUGHES, J.M. (1998). Evolution of Queensland spiny mountain crayfish of the genus Euastacus Clark (Decapoda: Parastacidae): preliminary 16s mtDNA phy- logeny. Proceedings of the Linnean Society of New South Wales 119, 9-19 The upland, mesic rainforests of Queensland are scattered in a series of disjunct mon- tane blocks along the east coast. They are remnants of a once widespread forest type that dominated Australia during the Miocene. Each of these montane blocks harbours a unique species of spiny mountain crayfish belonging to the genus Euastacus (Parastacidae: Decapoda). The preliminary 16s mtDNA data shows some incongruence between the phylogenet- ic relatedness and the geographic location of some species. To account for this a multiple range expansion-vicariance hypothesis is proposed. It is postulated that the group evolved from the range expansion of a southern ancestral form. This form underwent a range contrac- tion that resulted in at least two ancestral Queensland forms. These underwent a subsequent range expansion. A shift in climate, probably associated with the late Miocene drying of the continent, resulted in the ranges of these ancestral forms receding to upland areas. This isola- tion resulted in a large number of different species each with restricted distributions. Manuscript received 23 July 1997, accepted for publication 5 January 1998. Key words: Euastacus, freshwater crayfish, evolution, mtDNA, phylogeny. INTRODUCTION The upland rainforests of Queensland are scattered along the east coast in disjunct blocks. There is a relatively large, fragmented block on the mountain ranges in the south- east, a few small remnants on the isolated mountains of the central coast, and another rel- atively large, fragmented block on the mountain ranges in the Wet Tropics. The distribu- tion of these upland rainforest areas has been described as a ‘mesothermal archipelago’ as they constitute a chain of temperate mountain and tableland ‘islands’ that rise from a ‘sea’ of tropical and subtropical lowlands (Nix 1991). These mesic areas are remnants of a once more widespread forest type that dominated Australia during the early Miocene (Moritz et al. 1997 and references therein). It has been suggested that the historical expansion, contraction and fragmentation of these refugia played an important role in the speciation of rainforest restricted taxa (Heatwole 1987). Most of the mesic rainforest refugia of Queensland harbour a unique species of spiny mountain freshwater crayfish of the genus Euastacus (Morgan 1988, 1989; Horwitz 1990; Short and Davie 1993). There are now fifteen species described from Queensland (Short and Davie 1993). All but one, E. suttoni, which inhabits streams of the granite belt on the New England Tableland, inhabit the cool, fast flowing, well shaded streams of montane rainforests (Morgan 1988, 1989). This cold water adapted group (Riek 1969) is confined to higher altitudes as latitudes decrease. In south-east Queensland they occur only above 250 m, in central Queensland Proc. LINN. Soc. N.S.W., 119. 1998 10 16s mtDNA PHYLOGENY OF EUSTACUS above 750 m, and in north Queensland above 900 m (Morgan 1988, 1989; Cannon and Sewell 1994: 35, fig. 1). Riek (1959), noticing this interesting distribution, postulated that the group probably had a wide-ranging common ancestor that, once climatic conditions became hotter and drier, receded to the cooler mountains. This isolation resulted in a large number of different species each with restricted distributions. Morgan (1986, 1988, 1989, 1997), in the latest revision of the group, agreed that allopatric speciation was probably the dominant process of speciation. However, he stopped short of a detailed phylogeny or a detailed discussion of speciation. He identified only one morphological character, the presence or absence of a male cuticle partition, that may be useful in inferring phylogenetic relationships, although he was uncertain if this character was reliable. To understand the evolutionary diversification of this group it is imperative to have a robust phylogeny. The unique features of mtDNA which predispose it to evolutionary studies have been well documented (see Moritz et al. 1987; Avise 1994). This paper presents a preliminary 16s mtDNA phylogeny for eight of the fifteen species of Euastacus found in Queensland. We propose a hypothesis that may explain their evolution. Finally, we detail our current research which we hope, combined with a full phylogeny of the genus, will shed light on the evolution of these crayfish. MATERIALS AND METHODS Collection of Samples Samples of E. robertsi, E. fleckeri, E. balanensis, E. eungella, E. hystricosus, E. setosus, and E. sulcatus were collected from the field with a dip net (see Table 1), trans- ferred to liquid nitrogen for transportation, and once in the lab stored at —75°C. Specimens were identified using Morgan’s (1988, 1989) keys. Voucher specimens will be lodged with the Queensland Museum once this study is completed. Samples of E. suttoni were provided by Adrian Dawson. Tissue samples of E. spinifer were obtained from alcohol preserved samples from the Queensland Museum. Total genomic DNA of Cherax quadricarinatus was provided by Peter Mather, Queensland University of Technology. DNA Extraction, PCR Amplification and Sequencing Total genomic DNA was extracted by placing approximately 50 mg of muscle tis- sue in 350 | of extraction buffer (0.5 M Tris pH 8.0; 0.25 M NaCl: 0.025 M EDTA: 0.175 mM SDS) and | wl Proteinase K and incubated overnight at 55°C. Samples that were stored in alcohol were first soaked in STE for 10 mins. Proteins and lipids were removed by sequential extractions with equal volumes of phenol, phenol-chloroform-isoamy] alcohol (25: 24: 1) and chloroform-isoamy] alcohol (24: 1). DNA was precipitated at 4°C in 3 M sodium acetate (1/3 volume) and isoproponol (1 volume). Pellets were washed in 70% ethanol, dried and resuspended in 50 wl of TE buffer (10 Tris HCl, 1 mM EDTA, pH 7.5). The 16s ar and br primer set (Simon et al. 1994) was used to amplify a 461 base pair section of the 16s mtDNA fragment. PCR reactions contained 30 nMol each of dATP, dGTP, dCTP and dTTP (Promega), one unit of TAQ DNA polymerase (Promega), 2.5 mMol MgCl5 (Promega), 5 wl of 10 x polymerase reaction buffer (Promega), 0.5 uMol of each primer, 0.2 ug of template DNA and adjusted to a final volume of 50 1. DNA was initially denatured at 95°C for 5 min, then 35 cycles of 94°C denaturing for 1 min, 50°C annealing for 30 sec, and 72°C extension for | min, followed by a final 68°C extension step for 5 min. Proc. LINN. SOc. N.S.W., 119. 1998 M. PONNIAH AND J.M. HUGHES 1] TABLE | Collecting location of samples. Specimen Collecting Locality E. robertsi Hilda Ck., Thornton Peak E. fleckeri Paul’s Luck, Mt Spurgeon E. balanensis Kauri Ck., Lamb Range E. eungella Cattle Ck., Clarke Range E. hystricosus Bundaroo Ck., Conandale Range E. setosus Greens Falls, Mt Glorious E. sulcatus Canungra Ck., Lamington Plateau Three yl of the resulting PCR product was run on agarose gels and visualised using ethidium bromide. The remaining product was cleaned using Bresaclean (Bresatek), fol- lowing the manufacturer’s instructions. Sequencing reactions were carried out using dye terminator cycle sequencing reactions (Perkin Elmer) as per manufacturer’s instructions. 30-40 ng of cleaned template DNA and 3.2 pmol of primer were added. Sequencing was carried out on an Applied Biosystems 373 automated sequencing machine. Both the light and heavy strands were sequenced. Sequence Alignment and Phylogenetic Analysis Sequences for E. bispinosus, Cherax destructor albidus (formerly C. albidus, see Campbell et al. 1994) and Geocharax gracilis, published by Crandal et al. (1995), were incorporated into the analysis. Sequences were aligned with the program Clustal V (Higgins 1992). Genetic distances between sequences were calculated with Mega (Kumar et al. 1993) using the Tamura-Nei distance estimation model. Maximum likelihood analysis was performed using DNAML in Phylip (Felsenstein 1993) incorporating a transitions to transversions ratio of two. Unweighted maximum parsimony analysis was performed using the branch and bound search option in PAUP (Swofford 1993). Neighbour joining analysis was performed using Mega (Kumar et al. 1993) with a Tamura-Nei distance estimation model. Neighbour joining trees were constructed with gaps and missing infor- mation treated as both complete deletions and as pairwise deletions. Bootstrap values were calculated using 1,000 replications. G.-gracilis, C. quadricarinatus and C. d. albidus were used as outgroups. RESULTS The genetic differentiation between species is presented in Table 2. The phylogram generated by the maximum likelihood method, and geographic location of each species, is presented in Fig. |. It appears that the Queensland Euastacus examined are mono- phyletic (bootstrap 79%) to the exclusion of the southern species E. spinifer and E. bispinosus. Within this group there are three well supported clades: E. robertsi/E.bala- nensis (bootstrap 100%), E. eungella/E. setosus (bootstrap 70%) and E. hystricosus/E. sulcatus (bootstrap 74%). All three methods of phylogenetic analysis generated these clades. There is incongruence between the phylogenetic relatedness and geographic loca- tion of E. eungella and E. setosus. These two species clade together but occupy ranges which are geographically distant. Proc. LINN. SOC. N.S.W., 119. 1998 12 16s mtDNA PHYLOGENY OF EUSTACUS TABLE 2 Sequence divergences generated using Tamura-Nei distance estimation model. 2 3 4 5 6 7 8 9 10 11 1 13 ] 10277 0813" 0966 40930 .1028 0983 “1l6l “1741 2460) 2488) 22919 2 0737820946, 20967 20891) 21021-1091 OSIFy 216227 92253092323 2655 3 0421 LOGS OSA 05978 20616 0730 a3 8992677] ZO es Os 4 0643 .0414 .0698 .0672 .0749 .1268 .2756 .2382 .2466 5 0803 .0471 =.0652, 0835. 1449 2462) 2366) .2627 6 0746 .0651 .0596 .1405 .2632 .2377 = .2560 7 0577 =.0676 = 1417) 2430) 2269 .2633 8 0603" 21492" 2377 23337 2618 9 = OOfpee-2285 52252550) 10 2673 = .1762 = 2187 11 2671 = .3139 12 ; 2425 1. E. robertsi, 2. E. fleckeri, 3. E. balanensis, 4. E. eungella, 5. E. hystricosus, 6. E. setosus, 7. E. sulcatus, 8. E. suttoni, 9. E. spinifer, 10. E. bispinosus, 11. C. quadricarinatus, 12. E. C. d. albidus, 13. G. gracilis. DISCUSSION There are at least three hypotheses that can be erected to explain the evolution of the Queensland Euastacus; and because the mode and tempo of evolution will determine the shape of the phylogeny (Purvis 1996 and references therein) there are at least three topologically different hypothetical phylogenies. The first is a vicariance hypothesis. Once the ancestral Euastacus evolved it spread rapidly along the east coast of Australia during periods when favourable habitat pre- vailed (wet and cool) and when there were few barriers to dispersal. Once climate changed (warm and/or dry), the range of the ancestral Euastacus receded up the moun- tain ranges. Vicariance resulted in allopatric speciation. If this hypothesis is valid, one would expect neighbouring species to be more phylogenetically similar to each other than they are to geographically distant ones. Furthermore, if a shift in climate resulted in simultaneous vicariance events across the whole of the ancestral range, one would expect similar branch lengths leading to the tips of clades. There should be very few internodes, and for the internodes that are present, the branch lengths separating internodes should be very short. The tree should also be balanced. The second hypothesis is stepwise colonisation. Once the ancestral Euastacus evolved, there was a progressive stepwise colonisation and isolation. In this scenario, too, one would expect that there would be geographic and phylogenetic congruity. In this scenario, though, one would expect a well delineated basal clade, giving rise to another clade, which in turn is basal to another clade. The topology of the tree would not be symmetrical because the more derived clades would have much shorter branch lengths. The basal clades should correspond to where the ancestral Euastacus evolved. If the ancestral Euastacus evolved in southern Australia, one would expect the southern species to form the basal clades of the phylogeny and the northern species to be derived from them. Proc. LINN. SOC. N.S.W., 119. 1998 M. PONNIAH AND J.M. HUGHES 13 E. robertsi a 400 > wwe E, fleckeri 40 setserssnassvarsessaeethocee tae ee E. balanensis 52 E. eungella E. setosus 70 79 AE. hystricosus = Brisbane __ww E. sulcatus 98 47 E. suttoni 75 ww. Spinifer E.bispinosus C. quadricarinatus Melbourne G. gracilis Figure I. 16s mtDNA maximum likelihood phylogeny showing geographic location of each species on a schematic map. Neighbour joining bootstrap values have been included. In the third hypothesis the evolution of the group may be accounted for by multiple range expansions and vicariance events. The ancestral Euastacus once evolved, extends its range, then contracts leading to vicariance. These new forms extend their range when conditions are suitable, and then contract again when conditions are not favourable. In this scenario, one would not expect absolute congruence between geography and phylo- genetic relatedness. It is expected there would be well defined lineages that correspond to the periods of isolation. There should be some structure evident where closely related species would be geographical neighbours. Importantly, not all geographically neigh- bouring species would be closely related to each other. The most plausible of the three hypotheses seems to be the multiple range expan- sion-vicariance hypothesis because there is some incongruence between the phylogenetic relatedness and geographic location of some species. These hypotheses can not be tested because the 16s phylogram is generated from a preliminary data set and is not fully resolved. Proc. LINN. Soc. N.S.W., 119. 1998 14 16s mtDNA PHYLOGENY OF EUSTACUS There are, though, a few further points worth considering. First, E. bispinosus (a Victorian species) is the basal clade of the phylogram generated. It has long been postu- lated that southern Australia was the centre of origin of the Euastacus (Riek 1969). Second, the genetic distances between species indicate differentiation in the late Miocene or early Pliocene. Moritz et al. (1997) have outlined some of their work on the speciation of a number of endemic Queensland rainforest-restricted vertebrate taxa. Using the same fragment of 16s mtDNA, and assuming a 16s molecular clock that had genetic divergences of 1% roughly equating to 1 million years (C. Moritz pers. comm.), they tentatively postulated that speciation within these groups probably occurred during the Miocene or early Pliocene. If we apply the same clock to this data set (noting that the clock has not been calibrated nor developed for invertebrates), we get similar fig- ures indicating differentiation in the late Miocene or early Pliocene. These speciation events may coincide with the late Miocene contraction of mesic rainforests (Moritz et al. 1997). Interestingly, there is a Euastacus-like fossil Parastacid from the Eocene (Sokol 1987). Based on the preliminary 16s mtDNA data, a hypothesis can be erected to explain the possible evolution of the Queensland Euastacus. There was a range expansion of a southern Euastacus into Queensland. A subsequent range contraction resulted in at least two ancestral Queensland forms. These underwent a subsequent range expansion. A shift in climate, possibly the late Miocene drying, resulted in allopatric speciation of these ancestral forms. i The main reason for proposing a multiple range expansion-vicariance hypothesis for this group is that E. setosus, whilst being geographically closest to E. hystricosus and E. sulcatus, is phylogenetically closest to E. eungella. Another interpretation which may account for this incongruence is that the phylogram does not accurately reflect the evolu- tionary diversification of this group. Other genetic studies on crayfish (see Crandall and Fitzpatrick 1996; Crandall et al. 1995; and Avery and Austin 1996) found inconsistencies between the genetic and morphological data. In this case, however, the genetic data are consistent with Morgan’s (1988) morphological revision. We know very little about the dispersal capabilities of this group, et alone the dis- persal capabilities of any hypothetical ancestor. Merrick (1983) and Morgan (1997) have noted that the dispersal capabilities are probably very limited within this group. A few studies have tried to estimate the dispersal capabilities of species within this group. Geddes et al. (1993) used allozymes to infer gene flow within E. armatus. They found that this wide-ranging and, in some respects, unusual species (Morgan 1997), had a high level of genetic continuity across its range. Similarly, allozymes were used by Ponniah (1992) to infer the dispersal capabilities of E. hystricosus. However, he did not detect enough allozyme variation to be able to make any firm inferences. We are currently investigating the dispersal capabilities of four Queensland species of Euastacus (E. robertsi, E. fleckeri, E. hystricosus and E. sulcatus). Our study is focussing on the mode of dispersal (instream vs overland) and the types of habitats that are barriers to dispersal. We are using allozymes, mtDNA and the genetic population structure of the ectocommensal temnocephalans that inhabit these crayfish. Unpublished data for E. robertsi, which has three disjunct subpopulations, indicates that this species has very limit- ed dispersal capabilities between populations on different mountains, and even between streams on the same mountain which are not connected by continuous rainforests. We are also trying to construct a more robust mtDNA phylogeny by adding CO1 mtDNA sequence data. We will include the remaining species found in Queensland, incorporate sequence data from two individuals from each species, and incorporate a few more southern species so that the Queensland species may be put into context. With the development of a robust molecular phylogeny, the development of a reasonable molecu- lar clock, and a better understanding of the dispersal capabilities of select members with- in the group, more light may be shed on the evolution of this group. Proc. LINN. SOC. N.S.W., 119. 1998 M. PONNIAH AND J.M. HUGHES 15 ACKNOWLEDGEMENTS The help provided by Sonja Schmidt, Lewis Roberts, Chris Marshall and Malcolm de Zilva in collect- ing specimens is gratefully acknowledged. Craig Moritz and Chris Schneider made available their data on endemic vertebrates of Queensland and provided many valuable discussions on the evolution of these groups. Adrian Dawson provided samples of E. suttoni. Queensland Museum tissue samples of E. spinifer were provid- ed by John Short and Peter Davie. Total genomic DNA of Cherax quadricarinatus was provided by Peter Mather. Permits to collect were granted by the Queensland National Parks and Wildlife Service and by the Queensland Department of Forestry. This research was supported by a scholarship to M.P. and a grant from the Cooperative Research Centre for Tropical Rainforest Ecology and Management. Nick Campbell and Craig Moritz reviewed a draft copy of the manuscript and made many useful suggestions. REFERENCES Avery, L. and Austin, C.M. (1996). The biochemical taxonomy of the spiny crayfish genera Astacopsis and Euastacus in south eastern Australia. Records of the Museum of Victoria 56, 283-295. Avise, J.C. (1994). ‘Molecular Markers, Natural History and Evolution.’ (Chapman and Hall: London). Campbell, N.J.H., Geddes, M.C. and Adams, M. (1994). Genetic variation in Yabbies, Cherax destructor and C. albidus (Crustacea: Decapoda: Parastacidae), indicates the presence of a single, highly sub-structured species. Australian Journal of Zoology 42, 223-258. Cannon, L.R.G. and Sewell, K.B. (1994). Symbionts and Biodiversity. Memoirs of the Queensland Museum 36(1), 33-40. Crandall, K.A., Lawler, S.H. and Austin, C.M. (1995). A preliminary examination of the molecular phylogenet- ic relationships of some crayfish genera from Australia (Decapoda: Parastacidae). Freshwater Crayfish 10, 18-30. Crandall, K.A. and Fitzpatrick Jr., J.F. (1996). Crayfish molecular systematics: Inferences using a combination of procedures to estimate phylogeny. Systematic Biology 45, 1-26 Felsenstein, J. (1993). Phylip (phylogeny inference package) version 3.5c. Distributed by the author. Department of Genetics, University of Washington, Seattle. Geddes, M.C., Musgrove, R.J. and Campbell, N.J.H. (1993). The feasibility of re-establishing the River Murray crayfish, Euastacus armatus, in the lower river Murray. Freshwater Crayfish 9, 368-379. Heatwole, H. (1987). Major components and distributions of the terrestrial fauna. In ‘Fauna of Australia Volume 1A general articles’ (Ed. D.W. Walton) pp. 101-135. (Bureau of Fauna and Flora and Australian Government Publishing Service: Canberra). Higgins, D.G., Bleasby, A.J. and Fuchs, R. (1992). Clustal V: Improved software for multiple sequence align- ment. Caribos 8, 189-191. Horwitz, P. (1990). The conservation status of Australian freshwater crustacea with a provisional list of threat- ened species, habitats and potentially threatening processes. Report Series No. 14 Australian National Parks and Wildlife Service. Kumar, S., Tamura, K. and Nei, M. (1993). Mega: molecular evolutionary genetic analysis, version 1. The Pennsylvania State University, University Park, PA 16802. Merrick, J.R. (1993). ‘Freshwater crayfishes of New South Wales’. (Linnean Society of New South Wales: Australia). Morgan, G.J. (1986). Freshwater crayfish of the genus Euastacus Clark (Decapoda:Parastacidae) from Victoria. Memoirs of the Museum of Victoria 47(1), 1-57. Morgan, G.J. (1988). Freshwater crayfish of the genus Euastacus Clark (Decapoda:Parastacidae) from Queensland. Memoirs of the Museum of Victoria 49(1), 149. Morgan, G.J. (1989). Two new species of Euastacus Clark (Decapoda: Parastacidae) from isolated high country of Queensland. Memoirs of the Queensland Museum 27, 555-562. Morgan, G.J. (1997). Freshwater crayfish of the genus Euastacus Clark (Decapoda: Parastacidae) from New South Wales, with a key to all species of the genus. Records of the Australian Museum (1997) Supplement 23. Moritz, C., Dowling, T.E. and Brown, W.M. (1987). Evolution of animal mitochondrial DNA: Relevance for population biology and systematics. Annual Review of Ecology and Systematics 18, 269-292. Moritz, C., Joseph, L., Cunningham, M. and Schneider, C. (1997). Molecular perspectives on historical frag- mentation of Australian tropical and subtropical rainforests: implications for conservation. In “Tropical rainforest remnants — ecology, management, and conservation of fragmented communities’ (Eds W.F. Laurance and R.O. Bierregaard, Jr) pp. 442-454. (University of Chicago Press: Chicago). Nix, H.A. (1991). Biogeography — Patterns and processes. In ‘Rainforest animals: Atlas of vertebrates endemic to Australia’s wet tropics’ (Eds H.A. Nix and M. Switzer) pp. 11-39. (Australian National Parks and Wildlife Service: Canberra). Ponniah, M. (1992). The conservation genetics of Euastacus hystricosus Riek (Decapoda: Parastacidae). Unpublished Masters Dissertation, Faculty of Environmental Sciences, Griffith University. Purvis, A. (1996). Using interspecies phylogenies to test macroevolutionary hypotheses. In ‘New uses for new Proc. LINN. SOC. N.S.W., 119. 1998 16 16s mtDNA PHYLOGENY OF EUSTACUS phylogenies’ (Eds P.H. Harvey, A.J. Leigh Brown, J. Maynard Smith and S. Nee) pp.153-168. (Oxford University Press: Oxford). Riek, E.F. (1959). The Australian freshwater crustacea. In ‘Biogeography and Ecology in Australia’ (Eds A. Keast, R.L. Crocker and C.S. Christian) pp. 246-258. (Uitgevererij Dr. W. Junk: The Haig). Riek, E.F. (1969). The Australian freshwater crayfish (Crustacea: Decapoda: Parastacidae), with descriptions of new species. Australian Journal of Zoology 17, 855-918. Short, J.W. and Davie, P.J.F. (1993). Two new species of freshwater crayfish (Crustacea: Decapoda: Parastacidae) from northeastern Queensland rainforest. Memoirs of the Queensland Museum 34(1), 69-80. Simon, C., Frati, F., Beckenbach, A., Crespie., Liu, H. and Flook, P. (1994). Evolution, weighting, and phyloge- netic utility of mitochondrial gene sequences and a compilation of conserved polymerase chain reaction primers. Annals of the Entomological Society of America 87(6), 651-701. Sokol, A. (1987). A note on the existence of pre-Pleistocene fossils of parastacid crayfish. Victorian Naturalist 104, 81-82. Swofford, D.L. (1993). PAUP: Phylogenetic analysis using parsimony, version 3.1.1. Smithsonian Institute, Washington, D.C. Proc. LINN. SOC. N.S.W., 119. 1998 M. PONNIAH AND J.M. HUGHES 17 APPENDIX A Aligned sequences of the 16s mtDNA region for some members of the genus Eustacus and for Cherax quadricarinatus. 1 111111112 2222222223 3333333334 4444444445 1234567890 1234567890 1234567890 1234567890 1234567890 1(E. robertsi) GCCCATTG-G GTTACTAAAA GGCCGCGGTA TAGTGACCGT GCGAAGGTAG YU, ate oe Le wsban Peet te Mees eae he delsicip oy ici s oo, s9g 0.00890 B (Habalanensis) iiemew see ceils eee aa Dy ee Sew Ate. buoval os\1 wiso fq Wage BA Blige ol die.d ate, Boigea ic AGE GuAgala ap oe Gaan dle) so oGcagegoee C20 PED HB HuolG! Dio Doo 6 Oo mu bon Ga 6 on o S03, (OMIUGOMS)) cisco) a blo cdc Sale Geo 8 Soke Goo Rocio Glo ooo oD doc) oo lo God 6 1n e olo ic 6(E. setosus) ala Corts a gi aoa mage. iy Beat tar Um POST a ANT lest tale chetaorer ca c ratunyaeac AMS PRS ie Monn Me ates tera'G oly WCE, SRCTISY 3 S0oid Oho Seo ol ho oe ola we ad OP BA 6 BO Bo oo Ciple eo bo 6 6 5 ola %o 15 SUS, SHOR ee PSS ale Pee Sidhe SA! Blea oo lp'o eee 6 Ble 6 eho 6 Bee a eee 8 Bee OE, SDRATAD) ae Bleue ene oes Prams Ba velva seiisicnon ey ava Cece! Aho mtova I olin aes "bade i) tea Mic cleat 6 ow suatra ts 10(Cheraxquad) .......... Graham Grits ioe Clie Sa emceeremar ace HFN ee esi Ronni pea tetas cae ae agG 5555555556 6666666667 7777777778 8888888889 9999999990 1234567890 1234567890 1234567890 1234567890 1234567890 \(E. robertsi) CATAATCATT TGTCTTTTAA GGCCGCGGTA TAGTGACCGT TTGGACGAAA DN Be RATING Wl Sa i eeors Ae a hein oaks Re Bien, ay tone, eilenanelis Ue AUB EE NE Waans BAGS tor) iciaaa ole A 9 104 6) c 3(E. balanensis) .......... PA we eal at ee a are Ue culty ME TSree iE cine Mic sere NYRCON NEM METE, ch Ut AVE, QUIBAIO) “hg 6 6 6 66 4 a) A Nera a eae ge adel Neds te ate Soiasd, fle issues eh ethnic Awe wa Soares 5 (HRIVSTRICOSUS) Ines me) ace ee a AG Ue ero eee age SP PNW ap GON inca ane a ripe aM a WCDI OT 6 (ERISCLOSIS) IA eee ene) Oa IXGRAE NS MRE en Naa oee aye A rey HL cosh, a a ee a ge i (EMSULGALUS) ye ees INR ial ee aU AN NR Dal Sil soi rasp sl Se ad WR FAA AR G. SS (EMISTLELOTI leer se ee UNE earl ra eS PAU AT sat) Sa re Rt ol LR ONE er a VO THN pg GEL oe tse RA G. O(Eespinifen)\ «hea os cone / NES a ein dea eee Ny SAGAN a Rp Seer reat aiite ete, ghost en ais Cn aie G. 10(Cheraxquad) .......... PA ares Oat acre eters tea eet ee Gye Aveta ccs sam CSV aaa aa G. VID ITUI NT SMa a ei ie a ba at eae aed 0000000001 1111111112 2222222223 3333333334 4444444444 1234567890 1234567890 1234567890 1234567890 1234567890 1(E. robertsi) AGCGATCTGT CTCTA-TGGG AGTAATTGAA TTTAACTTTT GAGTGAAAAG PD (ERLECKETU)) an eurwlisten readers ta eaunsuetrrnce et SUANAR Aas Riek uMneyt) (via. een ay causes tas cid 2 Ue emeee T Be e 3(E. balanensis) ..T....... Saye leal Cited Cass VignvaNn SUB DeATO eee tee nme ss Ld eaoiues shat ra tore Ga we AGE, Grieg) %, UN's bicth 66 dau o's Dg NG. AVA WAY BR ecg ult nat Wes heey ty sal co lene ome Gc coh Gar S (ERB STTECOSUS) pmsl Noel ens ies Senay eee ee UY Tran GAVE NM olin Bein leases SPecas stoke" Oa og (alban rah bby 6(E. setosus) re Ne nena a ia Gr rshacqnil aA eet ONS ear yc cucarrtin ic ae cercaw. Lido se Manne a Nseyien G. 7 (EME SULGULES) (a ele Nee eee sR AN te NUA Gaerne cs Mae a ecg rapa dey oath enue inca alates Gee: 8(E. suttoni) Ry NS acne oo etna rate een TE NG ANU A Perl elie rearcatap Meee lanier, alley Suen amen se Grea Q(E. spinifer) ..TA...... ce oo Gea aN ceAVAbi5 do 3 Bla bo 6 odo a0 6 [Nin ata doe G IO (Gheraxquad)arw GA Gel ee eeele CIC GHNGws oe se Ys oe 8 8'8 6 7 Shee SMR Proc. LINN. SOC. N.S.W., 119. 1998 18 16s mtDNA PHYLOGENY OF EUSTACUS LUM a a ea Se ea hea i 5555555556 6666666667 7777777778 8888888889 9999999990 1234567890 1234567890 1234567890 1234567890 1234567890 1(E. robertsit) GCTTAAATGG TCTAGGGGGA CGATAAGACC CTATAAAGTT TAACATTATA 2 (ELECKCi) ee eee PN gene re a eease ig TY bi Were Ae 48 (an Ee aE Wk Wott eh vee ea co Re ea a S (EDGAR eNSIS) = eee Pa Cod Seer PTO eed eae pee ca en Oo ee eee 4(EVeungella) ys ee ae TANS eect ety ge INR, PARTE EK see BEI Aghc idee: (eR Srmice! ct Vie vos 8 dae ees » (ES StniGOSUS) Benen eae UNe lessee apace A ahi aa eee tepid eh peao eras ls any oF Selene ee Gus) SAGER G: OZ, SHOWS) soos coe 6s Nay eee ARSON Se RR eee ALR econ or ae ae Ne Renn 9 ee ((ESSULCAtUS) ie tonne ee eee ee) aes AGE Tah, AS Re En Cobre me be eh ee ee Ed a ee G Nid, SHIT) 6 ssoco0w abe Aue ee / ND LER EME! OL Ae AMI PEAT eee Ey, hie Oa Oe Gee EA MRM ON Ne, GOYA) cscoacnaac RES Toe PR ee Triad eae Bem eae as nN «>, SUL NO (Cheraxquad) re eee ee PACS Giles even hs hee Ninna PETE hrs ae eck eee 7 vACARGEe 7) 2b 0) 0) Py Dy ly Py ly Pyle, Ch D4 2 ADI IY Dy, Dy Py, 14 Ih Dy Cat) Oy Da Py Daly Dad AYA DADDDLZDD OOOO0OOOOOO!L TILI1111112 2222222223 3333333334 4444444445 1234567890 1234567890 1234567890 1234567890 1234567890 1(E. robertsi) GTAGTGAAAG ATTGATT--A AGT---TGTA GAATTTTTT-— ATTATGGGTA D(EaflCCKert) eee ene Gee eee AFAR Bee es Ae 2 = GG & Sica ls ns eee 3(E. balanensis) A. .A.A...A G..A GG. SO rea AG. A G AAA 4(E. eungella) A..ACA.T.A G..T GGGR = a ese A. AG. Ave: G..AA 5 (ES hyStricOSus)) sp A Awe eS (Gees ee eae Geer: G Geers AGe GCAAA 6(ESsetosus) WAC SACANGTARG WA 3GGG = —— eee C. A.GA Agu Gr aA (CES SULCATUS) em re re NO Arn ee NG Ope tee ee Gee roe GG) BG ACA Ss. sae an AAA. . 8(E. suttoni) Ave oA Ate S RAR Gee eer ney ate GREE aes AG. AG.AC. .A.T G.AA OUR Sauna) INC .NgINGa oN. oo uve o op c =1GwAs. 3. TAP ALGAAS cA. To. - AGRARAEG lO (Cheraxquad) ererilen Gills een Geer SAT i Grete aie GT.. A.G-...A.T .CC.GCA.-G DMX AZDDADM DDD, DAPI D DAA, DODD PDD ADA, D™PLAADADADMDDD DAP DMDDIDID §$555555556 6666666667 7777777778 8888888889 9999999990 1234567890 1234567890 1234567890 1234567890 1234567890 1(E. robertsi) GTGTTTTGTT GGGGCGACAA CGATAAGACC CTATAAAGTT TTCTTTTTTG PAO EVD CA TD Vem eer cians 800 m elevation), whereas in lower latitudes, they occur at sea level (Morgan 1988, 1991; Horwitz 1990; Short and Davie 1993; Cannon and Sewell 1994). By contrast, few Australian species of Cherax [a genus restricted to mainland Australia, southern New Guinea including the Aru Islands and Misool [Palau I.] (Reik 1969, 1972; Banarescu 1990; Short and Davie 1993)], are endemic to wet upland or highland areas: the crayfish are found commonly in slower, lowland or coastal streams and in the foothills of uplands, often (but not always) with open canopy above (Riek 1969; Short and Davie 1993; Cannon and Sewell 1995). In north and mid-eastern Queensland, populations of Cherax and Euastacus are separat- ed by pronounced physical barriers and, with the single known exception of Cherax parvus which occurs sympatrically with Euastacus yigara in the Tully River catchment (at an altitude of c. 750 m), are clearly discrete (Morgan 1988,1991; Horwitz 1990; Short and Davie 1993). As latitude increases, however, Euastacus are less confined to high altitude, and in southern Australia, suitable habitat for Euastacus occurs down to sea level (Morgan 1988; Cannon and Sewell 1994). Thus, in southern Australia there exists potentially more opportunity for ectosymbiotic worms such as the Craspedellinae to ‘host switch’ between Cherax and Euastacus crayfish. Of the two species of Craspedellinae known to infect Euastacus, i.e. Gelasinella powellorum and Heptacraspedella peratus, both occur at moderately high latitudes, in mid-eastern NSW, and in the Grampians region, respectively. In the Grampians, which is the type locality of H. peratus, both Euastacus and Cherax were reported by Riek (1972) to co-inhabit streams. The depauperate distribution of Craspedellinae on Euastacus and particularly their apparent absence from the ‘relic’ Queensland populations of the genus suggests that the Craspedellinae have not radiated with these hosts although the possibility of extinctions in Euvastacus populations cannot be excluded. Cannon and Sewell (1995) proposed that the Craspedellinae may not be detected in populations of Euastacus if only small cray- Proc. LINN. SOc. N.S.W., 119. 1998 K.B. SEWELL AND L.R.G. CANNON 35 fish were examined. They suggested this to be the reason that Craspedellinae were not found in their samples of mainly small Euastacus collected from Cape York to the Grampians. This argument is not supported, however, by data from the Karuah River region, NSW. On 21/Nov/1996, KBS and Dr Robert D. Adlard found numerous Gelasinella powellorum on a small juvenile (25 mm occipital carapace length (OCL)) as well as on a large adult (83 mm OCL) specimen of Euastacus spinifer collected from Mammy Johnsons Creek, just south-east of Stroud Road township, NSW (32°20.57’S; lesiligs onluliils): Queensland species of Euastacus do not appear to be hosts to Craspedellinae: none of 13 Queensland populations of Euastacus, including six ‘relic’ ones, examined for tem- nocephalans during the course of fieldwork by Cannon and Sewell (1994), was infected with Craspedellinae and examination of small and large specimens collected subsequent- ly from southern and northern Queensland also failed to reveal any species. The Craspedellinae thus probably have radiated with Cherax as their hosts and have trans- ferred subsequently to Euastacus only in southern regions where the host genera may/can occur sympatrically. It is pertinent to note, however, that EF. yigara and C. parvus, which occur sympatrically in northern Queensland, have not yet been examined for Craspedellinae. Craspedellinae are unknown from Tasmanian parastacids. Craspedellinae were not reported from any Tasmanian crayfish by either Haswell (1893) or Hickman (1967), nor have they been detected subsequently (Cannon and Sewell unpublished observations). The absence of the worms is consistent with the fact that neither Cherax nor Euastacus occur in Tasmania (Riek 1969; Holthuis 1982). Euastacus also are absent from Papua New Guinea, although numerous species of Cherax are recorded there (Holthuis 1949, 1982). Therefore, the report of a putative species of Craspedellinae from Cherax commu- nis in Papua New Guinea by Cannon and Sewell (1995), further supports the hypothesis that the Craspedellinae have radiated with Cherax. The biogeographical distribution of the Craspedellinae is consistent with an origin in Australia/New Guinea after the separa- tion of South America and Australia from Antarctica (c.45mya). ACKNOWLEDGEMENTS We thank the Australian Biological Resources Study for a grant (87/5909) to LRGC which partly fund- ed the study. The support of the Queensland Museum (QM) and The University of Queensland is also grateful- ly acknowledged. Mr John Short of the Crustacean Section, QM cheerfully supplied expert identification of crayfish, literature on the hosts and assistance in the field. Mrs Zeinab Khalil, of the Worms Section QM pro- vided invaluable and cheerful assistance with histological serial section preparation. We thank Mr Peter Davie of the Crustacean Section, QM for valuable comments on an early draft of this manuscript. Euastacus spinifer collected by Powell J. and Powell R. and by KBS and Dr R. D. Adlard were collected under NSW Fisheries permit numbers F95/346 and F96/363 respectively. REFERENCES Banarescu, P. (1990) ‘Zoography of fresh waters’. Volume 1. General Distribution and Dispersal of Freshwater Animals. 511 pages (Aula-Verlag: Wiesbaden). Cannon, L.R.G. (1993). New temnocephalans (Platyhelminthes): ectosymbionts of freshwater crabs and shrimps. Memoirs of the Queensland Museum 33, 17-40. Cannon, L.R.G. and Sewell, K.B. (1994). Symbionts and biodiversity. Memoirs of the Queensland Museum 33, 36-40. Cannon, L.R.G. and Sewell K.B. (1995). Craspedellinae Baer, 1931 (Platyhelminthes: Temnocephalida) ectosymbionts from the branchial chamber of Australian crayfish (Crustacea: Parastacidae). Memoirs of the Queensland Museum 38, 397-418. Cannon, L.R.G. and Watson N.A. (1996). Postero-lateral glands of Temnocephala minor Haswell, 1888 (Platyhelminthes: Temnocephalida). Australian Journal of Zoology 44, 69-73. Proc. LINN. SOC. N.S.W., 119. 1998 36 NEW TENNOCEPHALANS FROM CRAYFISH Evans, G.O., Sheals, J.G. & MacFarlane, D. (1961). ‘The Terrestrial Acari of the British Isles — An Introduction to their Morphology Biology and Classification’. Volume 1. Introduction and Biology. 219 pages (Adlard & Son, Bartholomew Press: Dorking, Great Britain). Haswell, W.A. (1893). A monograph of the Temnocephaleae. Linnean Society of New South Wales, Macleay Memorial Volume, 93-152. Hickman, V.V. (1967). Tasmanian Temnocephalidea. Papers and Proceedings of the Royal Society of Tasmania 101, 227-250. Holthuis, L.B. (1949). Decapoda Macrura, with a revision of the New Guinea Parastacidae. Zoological results of the Dutch New Guinea expedition 1939. Nova Guinea 5, 289-328. Holthuis, L.B. (1982). Freshwater Crustacea Decapoda of New Guinea. In: Biogeography of New Guinea. Monographiae Biologicae 42, 603-619. Horwitz, P. (1990). The conservation status of Australian freshwater crustacea with a provisional list of threat- ened species, habitats and potentially threatening processes. Australian National Parks and Wildlife Service Report Series Number 14. 121 pages. Joffe, B.I., Solovei, I.V. and Cannon, L.R.G. (1995a). The structure of the epidermis in Didymorchis (Temnocephalida: Platyhelminthes). Australian Journal of Zoology 43, 631-641. Joffe, B.I., Solovei, I.V., Sewell, K.B. and Cannon, L.R.G. (1995b). Organisation of the epidermal syncytial mosaic in Diceratocephala boschmai (Temnocephalida: Platyhelminthes). Australian Journal of Zoology 43, 509-518. Morgan, G.J. (1988). Freshwater crayfish of the genus Euastacus Clark (Decapoda, Parastacidae) from Queensland. Memoirs of the Museum of Victoria 49, 1-49. Morgan, G.J. (1991). The spiny crayfish of Queensland. The Queensland Naturalist 31, 29-36. Riek, E.F. (1969). The Australian freshwater crayfish (Crustacea: Decapoda: Parastacidae) with descriptions of new species. Australian Journal of Zoology 17, 855-918. Riek, E.F. (1972). The phylogeny of the Parastacidae (Crustacea: Astacoidea), a description of a new genus of Australian freshwater crayfishes. Australian Journal of Zoology 20, 369-389. Sewell, K.B. and Cannon, L.R.G. (1995). A scanning electron microscope study of Craspedella sp. from the branchial chamber of redclaw crayfish, Cherax quadricarinatus, from Queensland, Australia. Hydrobiologia 305, 151-158. Sewell, K.B. and Whittington, I.D. (1995). A light microscope study of the attachment organs and their role in locomotion of Craspedella sp. (Platyhelminthes: Rhabdocoela: Temnocephalidae), an ectosymbiont from the branchial chamber of Cherax quadricarinatus (Crustacea: Parastacidae) in Queensland, Australia. Journal of Natural History 29, 1121-1141. Short, J.W. and Davie, P.J.F. (1993). Two new species of freshwater crayfish (Crustacea: Decapoda: Parastacidae) from northeastern Queensland rainforest. Memoirs of the Queensland Museum 34, 69-81. Williams, J.B. (1982). Studies on the epidermis of Temnocephala V1. Epidermal topography of T. novaezealan- diae and other Australasian Temnocephalids, with notes on microvillar and coated vesicle function and the evolution of a cuticle. Australian Journal of Zoology 30, 375-390. Williams, J.B. (1992). Scanning electron microscopical study of Temnocephala fasciata (Platyhelminthes: Temnocephaloidea). Journal of Submicroscopic Cytology and Pathology 24, 205-213. Proc. LINN. SOC. N.S.w., 119. 1998 Egg and Juvenile Development of the Australian Freshwater Crayfish, Euastacus bispinosus Clark (Decapoda; Parastacidae) JODIE A. HONAN (Communicated by J. Merrick) Box 205, Port Fairy, Victoria 3284, Australia (formerly School of Aquatic Science and Natural Resources Management, Deakin University, PO Box 423, Warrnambool, Victoria 3280) Honan, J.A. (1998). Egg and juvenile development of the Australian freshwater crayfish, Euastacus bispinosus Clark (Decapoda; Parastacidae). Proceedings of the Linnean Society of New South Wales 119, 37-54. Aspects of egg attachment, embryogeny and development of early juvenile stages were investigated in Euastacus bispinosus from south-west Victoria. Shortly after spawning the eggs (4.0 x 2.9 mm) become attached in small groups to filamentous oosetae on the maternal pleopods. The egg nauplius takes two weeks to develop and at 90 days an egg mysis has formed, with eye pigmentation occurring at about 100 days. The development of eggs takes about 140 days. Hatching takes 30-60 minutes and the embryo moults at the same time. The cast exoskeleton forms a temporary link to the mother’s pleopod by adhering to the telson for 24-36 hours. The Stage 1 juveniles then grip the pleopods with their fourth and fifth pereiopods. After two more moults juveniles become independent at about 170 days. Euastacus bispinosus development is similar to other parastacids and key features are compared with those of other crayfish groups. Possible evo- lutionary relationships are also briefly discussed. Manuscript received 23 July 1997, accepted for publication 19 November 1997. KEYWORDS: Egg attachment, embryogeny, Euastacus bispinosus, juvenile stages, reproduction. INTRODUCTION As part of broader studies on reproductive cycles, growth and management of the Glenelg River crayfish, Euastacus bispinosus (Honan and Mitchell 1995a, b, c), observa- tions were made on egg attachment, embryo development and early juvenile stages. This large species is endemic to the Glenelg River system (south-west Victoria) and coastal drainages of south-eastern South Australia (Morgan 1986). Egg and juvenile development have been described for many freshwater crayfish (Hamr 1992). Eggs of northern and southern hemisphere species develop in similar ways, but juveniles differ in their method of attachment to the maternal pleopods (Gurney 1935, 1942) which indicates independent invasions of fresh water. The eggs and juveniles of many Australian genera have been described, but there is no full description for any Euastacus species. Clark (1937) and Turvey and Merrick (1997) include partial descriptions. Euastacus 1s a widespread genus endemic to the east, south-east and south- ern coastal areas of mainland Australia (Morgan 1997). Objectives of this paper are: to describe the setal attachment of eggs to the maternal pleopods; to document embryo and larval development in the egg; to describe hatching and major changes in juvenile stages up to release; to compare and contrast developmental features of the species with other freshwater crayfish groups, and con- sider the evolutionary relationships between the superfamilies. Proc. LINN. Soc. N.S.W., 119. 1998 38 EGG AND DEVELOPMENT OF EUASTACUS METHODS Three egg-bearing females from the Glenelg River drainage were kept in 720 L stainless steel vats with near natural photoperiod and temperature changes (although the temperatures were about 5°C warmer than in the field). The warmer temperatures were likely to speed up development, but not alter the sequence (Wear 1974). They were fed a varied diet, and the water was changed monthly. Eggs or juveniles were removed from the females monthly, then weekly, then daily as hatching approached. The colour, texture and movement of live eggs and juveniles were noted and photographs were taken. Samples were preserved in 10% formalin and transferred to 90% ethanol. To observe hatching, a group of advanced eggs was carefully cut from a pleopod of one female and maintained in dechlorinated, aerated tap water. These eggs were checked frequently. Two or three eggs or juveniles were removed from each female captured during monthly sampling in south-west Victoria (Honan and Mitchell 1995a). The samples were preserved in alcoholic Bouin’s fluid and transferred to 70% ethanol after 24-48 hours (after Johnson 1979). This technique discoloured the eggs and juveniles, and distorted the cara- pace of juveniles. The preserved material was however valuable for examining setation and other fine detail and for following development in the field. Illustrations were constructed from a combination of photographs and sketches of fresh and preserved material. RESULTS The pleopods of E. bispinosus have an endopodite and exopodite of similar shape and size. They have three types of setae which are similar to those described by Thomas (1970). Pinnate setae (~4.6 mm long) fringe the margins of the exopodite and endopodite. Filamentous oosetae (~5.1 mm long) are only found on mature females. These setae grow in groups of 10-18 along all margins of the endopodite, except the tip. They also occur on the basal third of the outer edge of the exopodite, and mesially on the basipodite. Serrulate setae are short (0.5—1.2 mm) and sturdy and are found on the surface of the pleopods, rather than the margins. In males serrulate setae occur on the outer edges of the anterior face of the endopodite and exopodite, and on the posterior face of the exopodite. In females serrulate setae occur along all margins of the posterior faces of both podites. Most eggs are attached to filamentous oosetae on the maternal pleopods (Fig. 1). On large females, eggs may also rarely attach to setae at the ventral junction of the abdomen and cephalothorax. Each egg is surrounded by a balloon-shaped membrane, with a twisted stalk at one end. The stalk is not in a constant position relative to the developing embryo. Groups of fifteen or more filamentous oosetae are gathered and their tips held twisted together by the egg stalk. Up to ten eggs have their stalks joined to a large twisted bunch of setae, although groups of two or three, or single eggs are more common. Occasionally the stalk envelops pinnate setae and produces a matted area rather than a distinct twisted cord. Some eggs have no stalk, instead the setae adhere to the side of the egg. Incubation in the field lasted 18—20 weeks; eggs were spawned in early May, and hatched in October-November. Eggs in the laboratory were about 5°C warmer, and hatched about four weeks earlier. Egg-bearing females feed during incubation and are easily caught in the field (Honan and Mitchell 1995a). The approximate ages are for eggs developing in the field. The eggs are pale burgundy, soft and elongate (4.0 x 2.9 mm) shortly after spawn- ing. The first stage observed was an early gastrula with well developed blastopore. This invagination is about 200 um in diameter. Surrounding the pore is a slightly raised area about 750 um in diameter. Two days later the pore is much deeper (Fig. 2a). The rudi- ments of the protocerebra and the two thoracico-abdominal segments have started to develop. One day later the blastopore has begun to close over, forming an advanced gas- trula (Fig. 2b). The embryonic area has differentiated into the cephalic regions, and the Proc. LINN. Soc. N.S.W., 119. 1998 J.A. HONAN 39 PS FS omen ae a St Figure 1. Attachment of eggs to the maternal pleopods of E. bispinosus: E, egg; FS, filamentous ooseta; Pl, pleopod; PS, pinnate seta; St, egg stalk. Proc. LINN. Soc. N.S.W., 119. 1998 40 EGG AND DEVELOPMENT OF EUASTACUS rudiments of the first and second antennae. The thoracico-abdominal plate is fused, much thicker and semicircular, partially covering the pore. The pore takes at least four days to close over, but the mechanism was not observed. Large oil globules in the space between the yolk and the outer membrane are a feature of this stage. The egg-nauplius is about 500 wm long and takes about fifteen days to develop (Fig. 2c). The protocerebra are visible, but the cephalic lobes are very faint. The first antennae, second antennae, mandibles, cephalic and thoracico-abdominal rudiments have developed, and the blastopore is covered by the large thoracico-abdominal rudiment which has an anal involution. The mouth is bordered by the labrum and metastoma. A few small oil globules are present. At 30 days (Fig. 2d) the embryo is about | mm long. The cephalic lobes and proto- cerebra have changed little. Faint grooves have developed on the appendages indicating future segmentation. Three pairs of ganglia associated with the antennae and mandibles ee ee eocece ee e e cry hd Pee ee °, Bye oh ThAb ° Py b eo oe? 'e e? Pe EO aie on r Ay 2 ee _____py 250um | °° . ° ee Yt dda Figure 2. Development of E. bispinosus embryo, all views ventral, except (1), lateral. Ages are approximate: (a), several days after fertilisation; (b), showing partially closed blastopore; (c), 15 days; (d), 30 days; (e), 60 days: (f), 75 days; (g), 90 days; (h), 100 days, (i), 120 days. Abbreviations: Al, A2, first and second antennae; Ab, abdomen; An, anus; BI, blastopore; Br, branchiostegite; Ca, carapace; Ce, cephalic lobes; CF, caudal furca; De, deutocerebrum; E, eye; EM, egg membrane; ES, eye stalk; G, ganglion; H, heart; Hel and He2, dorsal and lateral lobes of hepatopancreas; Lab, labrum; Me, metastoma; Mn, mandible; Mng, mandibulary ganglion; MnP, mandibulary palp; Mo, mouth; Mx1, Mx2, first and second maxillae; Mxp1 to Mx3, first to third maxil- lipeds: P1 to P5, first to fifth pereiopods; P11 to P14, first to fourth pleopods; Pr, protocerebrum; Ret, retina; Ro, rostrum; St, stalk; Th, thorax; ThAb, thoracico-abdominal rudiment; Tr, tritocerebrum; Y, yolk. Proc. LINN. SOC. N.S.W., 119. 1998 J.A. HONAN 4] Ai a ome A "OG. ° e 5? OF a 95 e hee ° ° ° ° Opn ° e O 6 e S a af. s QO 2 4 ° C6 : x Sec o Ste 3 — SSS ° ° . e ° ° ° . ° Bree ret aos . e : es pte" 50. Gracias : ° . 5 X no's og Bo 14 of 6 5 si ff ve 7 ° : “ e ° z 5 be BY Os 5 e Cae 0 vs “te % . e oa) ° On oe? e . « . e e we ° ° . ° ° ery ° - a O 5 02 e S ane . . 4 0010 . 5 “es ry 2 * ,e e e . , : - ° fers . D , e Ri an 50 O ° . ° ° ° ° . ° os ae re. re 5 xe ME Sh 100um . eee : i ) 0 ° O 9-5 ier O nn) . aloha Neen we ee Figure 2 continued. Development of E. bispinosus embryo, (b), showing partially closed blastopore. Proc. LINN. SOc. N.S.W., 119. 1998 42 EGG AND DEVELOPMENT OF EUASTACUS Figure 2 continued. Development of E. bispinosus embryo, (c), 15 days; (d), 30 days. Proc. LINN. SOC. N.S.W., 119. 1998 Figure 2 continued. Development of E. bispinosus embryo, (e), 60 days; (f), 75 days. J.A. HONAN 43 Proc. LINN. SOc. N.S.W., 119. 1998 44 EGG AND DEVELOPMENT OF EUASTACUS Figure 2 continued. Development of E. bispinosus embryo, (g), 90 days. Proc. LINN. Soc. N.s.W., 119. 1998 J.A. HONAN 45 750um if St EM Ro E Hel H Al He2 A2 Br PI Mxp3 Ab P5 Figure 2 continued. Development of E. bispinosus embryo, (h), 100 days, (i), 120 days. Proc. LINN. Soc. N.S.W., 119. 1998 46 EGG AND DEVELOPMENT OF EUASTACUS surround the labrum which has grown, covering the mouth and metastoma. The first and second maxillae are forming. The thoracico-abdominal rudiment is still large and simple with a central anus and is faintly rimmed by the branchiostegites. The 60 day embryo (Fig. 2e) has well defined cephalic lobes with a distinct stalk. The first and second antennae have lengthened, but only the second antennae are bifur- cate. The maxillae and maxillipeds form a row either side of the mid line and appear bilobed. The thoracico-abdominal rudiment has lengthened, growing over the ventral surface towards the labrum, and is fringed by the pereiopods and pleopods. These appendages are simple without segmentation. The branchiostegites are well developed and the rostrum is forming between the eyes. The embryo has increased to 1.4 mm in length by 75 days (Fig. 2f). Differentiation of the eyes and eye stalks has continued. Both pairs of antennae are now bifurcate and are clearly segmented. The endopodites of the second antennae have lengthened. The mandibles, maxillae and maxillipeds are still simple, rounded rudiments. The bran- chiostegites form a thick rim around the embryo. The thoracico-abdominal rudiment is differentiated into thoracic and abdominal regions and the thoracic region has moved backwards beneath the carapace. The pleopods are concealed beneath the partially seg- mented abdomen. The pereiopods are elongate and faintly segmented. At 90 days (Fig. 2g) an egg-mysis has formed. The mouthparts have lost their sim- ple rounded appearance, and begun to differentiate. The segmented pereiopods emerge from beneath the branchiostegites and the first pair have developed chelae. The thoracic region has moved up under the carapace. The abdomen is partially obscured. The ros- trum is faint. At about 100 days the eye pigmentation and orange hepatopancreas appear (Fig. 2h). The hepatopancreas is preceded by deep furrows in the egg yolk. By 110 days the hepatopancreas consists of two bilateral pairs of organs. The heart is steadily beating and the intestine can be seen through the abdomen. By 120 days (Fig. 21) the embryo is very similar to just before hatching. The well developed eye stalks are crowned by pigment, the pereiopods are segmented as are the first antennae. The rostrum appears calcified. The abdomen is clearly segmented and the caudal furca have fused. From this point only the relative proportions and positions of appendages and organs change. Just prior to hatching the appendages are tightly packed. Some specimens have distal spines on the second and third chelipeds. The carapace is pigmented red. Rotifers, ciliates and a platyhelminth were noted moving over eggs and may have fed on matter that accumulated on the outside. Eggs hatched at about 140 days. For several days before hatching the embryo becomes more active. The “yolk” and pereiopods move occasionally and the scaphog- nathite beats infrequently. During its long development, the egg diameter increases only 5-10%. The hatching juvenile sheds the outer egg shell and a close fitting embryonic cuti- cle. The embryo thus hatches and moults at the same time. The egg shell splits suddenly along a longitudinal dorsal plane and the juvenile crayfish hatches head first (Fig. 3). Hatching takes about 30-60 minutes during which the heart and scaphognathite continue to beat. Some crayfish had difficulty hatching. Some came out abdomen first and could not free themselves from the shell. Others caught their legs in the cast exoskeleton. These crayfish were deformed or died. After hatching, the crayfish stretches, and hangs suspended by a telson connection for several minutes while the exoskeleton hardens. The crayfish then becomes very active, and after a few minutes it grasps the egg shell or stalk with the fourth and fifth pereiopods. The shed exoskeleton and cast lining of the intestine form the telson connec- tion that breaks after 24—36 hours. Stage | juveniles have a bulbous cephalothorax and a telson with no uropods, setae or setal buds (Fig. 4a). The yolk is a deep burgundy and the hepatopancreas bright orange. The rostrum is depressed between sessile eyes. The gills beat beneath the well Proc. LINN. SOC. N.S.W., 119. 1998 J.A. HONAN 47 Figure 3. Hatching of E. bispinosus juvenile: (a) shell just split, (b) dorsal sur- face free, (c) pulling anterior free, and (d) attached only by telson connection. calcified branchiostegites. The first and second antennae curve back beneath the body. All appendages other than the second maxillae lack setae. Late Stage 1 juveniles are more pigmented, more elongate and occasionally extend their abdomens. They grip the mother’s pleopodal setae using hooks on the dactyli of the fourth and fifth pereiopods. After 7 days the juveniles moult to Stage 2. Moulting is similar to that of an adult crayfish and takes 15-20 minutes. After moulting the juvenile hangs suspended by the cast lining of its intestine for about ten minutes before it grips the maternal setae with the 4th and 5th pereiopods and the chelae of the third pereiopods. Stage 2 juveniles have well developed setae, body sculpture and pigmentation (Fig. 4b). The telson still lacks setae, but has 18—20 setal buds. The pereiopods are much longer and thinner than those of stage 1. The abdomen is much larger. The carapace is sculptured by grooves. For 2—3 days prior to moulting white gastroliths are visible through the carapace. About 24 days after hatching the juveniles moult to Stage 3, and resemble minia- ture adults with a complete tail fan, numerous setae and more pigmentation (Fig. 4c). The yolk is much reduced. Initially they continue to cling to the mother and each other. Proc. LINN. SOC. N.S.W., 119. 1998 48 EGG AND DEVELOPMENT OF EUASTACUS Oa Tr < Eee sy Q & cece -~ fee eco” o x He2 Al AS Mxp3 A Pl Figure 4. Juvenile stages of E. bispinosus after hatching: (a), Stage 1, one day after hatching; (b), Stage 2; (c), Stage 3. Abbreviations as per Figure 2. Also AS, antennal scale; BrG, branchiocardiac groove, CeG, cephalic groove; Gi, gills; Sc, scaphognathite; Ssp, suborbital spine; Te, telson; U,uropod. Proc. LINN. SOc. N.S.W., 119. 1998 Imm J.A. HONAN 49 Figure 4 continued. Juvenile stages of E. bispinosus after hatching. Proc. LINN. SOc. N.S.W., 119. 1998 EGG AND DEVELOPMENT OF EUASTACUS 50 wAOOslL Figure 4 continued. Juvenile stages of E. bispinosus after hatching: (c), Stage 3. N.S.W., 119. 1998 Soc. Proc. LINN. J.A. HONAN 51 Within 48 hours of moulting, juveniles begin to feed. Juveniles become fully indepen- dent after about a week as they lose their yolks and clinging instincts. Initially juveniles eat egg cases, exoskeletons and dead juveniles, then catch zooplankton and graze sessile algae. They moult to Stage 4 after a further 3 weeks. DISCUSSION The eggs of decapods are usually attached to long, smooth, filamentous oosetae on the maternal pleopods (Gurney 1942). There has been confusion about the pleopodal setae. Both pinnate setae (Thomas 1970; Turvey and Merrick 1997) and filamentous oosetae (Hopkins 1967; Morrissy 1970; Suter 1977) have been called “plumose setae”. The pinnate setae and filamentous oosetae of E. bispinosus are similar to those of other parastacoids (Hopkins 1967; Turvey and Merrick 1997) and astacoids (Thomas 1970). However Andrews (1907b) describes egg-bearing setae of Orconectes as more like a bottlebrush than a feather. Serrulate setae were illustrated by Hopkins (1967) on the pleopods of Paranephrops planifrons but have not been described from the pleopods of other freshwater crayfish. The Parastacoidea tend to have narrower pleopodal podites with less basal dilation and more filamentous oosetae that may form dense fringes rather than tight bunches (Hopkins 1967; Johnson 1979; Turvey and Merrick 1997). In the Astacoidea and Nephropoidea the exopodite is larger and has a basal dilation (Fig. 5), perhaps to aid in ich A B C Figure 5. Distribution of filamentous oosetae on the pleopods of female astacurans in breeding condition: (a), Homarus vulgaris (Yonge 1937); (b), Austropotamobius pallipes (Thomas 1970); (c), Euastacus spinifer (Turvey and Merrick 1997). Diagrammatic, other types of setae not illustrated. Proc. LINN. SOc. N.S.W., 119. 1998 a EGG AND DEVELOPMENT OF EUASTACUS swimming. Burrowing parastacids may have even denser oosetae (Suter 1977), but the setation of the Cambaridae is unclear. Euastacus bispinosus has a similar egg stalk to other parastacoids (Hamr 1992; Turvey and Merrick 1997) and astacoids (Thomas 1991). The egg stalk of some species is in a constant position relative to the embryo (Johnson 1979; Stko 1962), but in E. bispinosus, Astacopsis and Parastacoides (Hamr 1992) this is not the case. The morphological changes in the embryo of E. bispinosus are similar to those of other freshwater crayfish (Hamr 1992; Johnson 1979; Reichenbach 1877, cited in Huxley 1880; Reichenbach 1886, cited in MacBride 1914; Sandeman and Sandeman 1991). Both Cherax destructor (Sandeman and Sandeman 1991) and E. bispinosus take 10—15 days to reach egg-nauplius, despite very different incubation periods (40 versus 140 days). As development continued the larger E. bispinosus eggs developed more slowly. Euastacus bispinosus eggs continued to develop during winter, whereas in the Astacidae a winter diapause of 3—4 months occurs when low water temperatures inhibit development (Cukerzis 1988). During development the egg diameter of E. bispinosus increased only 5—10%. The eggs of Astacoidea may increase by 13—27% (Hessen et al. 1987; Koksal 1988; Mason 1977). The eggs of Procambarus clarkii and Astacus astacus increase their diameters by 16-19% in the one or two days before hatching (Hessen et al. 1987; Stko 1962). Euastacus bispinosus split the egg dorsally and hatched legs last. Clark (1937), however, noted that Euastacus split the egg with its abdomen and rapidly hatched legs first. It is unlikely that members of the same genus would have very different methods of hatching. A few E. bispinosus did attempt tail first hatching, but it was slow, and defor- mities and mortality were high as the crayfish shed an exoskeleton as well as an egg shell. Freshwater crayfish typically hatch and moult at the same time (Andrews 1904, 1907a, 1907b; Gurney 1942; Hamr 1992; Sandeman and Sandeman 1991). In E. bispinosus the dorsal split is along the axis of the animal whereas in the Astacidae it seems to be across the axis of the animal (Andrews 1904, 1907b; Stko 1962). In both the Astacoidea and Parastacoidea telson connections join newly hatched juveniles to their shells (Rudolph and Rios 1986; Rudolph and Zapata 1986), and may break soon after hatching or persist for several days (Andrews 1907a, 1907b; Gurney 1942; Hamr 1992; Holdich and Reeve 1988; Hopkins 1967; Johnson 1979; Stko 1962; Turvey and Merrick 1997). The connection of E. bispinosus lasts 24—36 hours; the juve- niles then grip with their fourth and fifth pereiopods. In both the Parastacoidea and Astacoidea the connection is formed from the cast embryonic cuticle. In E. bispinosus and other Parastacoidea the embryonic cuticle adheres to the telson which has a smooth edge (Hamr 1992; Hopkins 1967; Sandeman and Sandeman 1991). In the Astacoidea the membrane is thought to be attached by secretions from the setal buds or spines on the outer edge of the telson (Andrews 1907b). The telson connection may give way so that the only link between the juvenile and the mother is an anal thread, the cast lining of the gut. This thread lengthens with time as more is pulled free with the weight of the juvenile (Andrews 1907b). The Stage | juveniles of E. bispinosus are typical of freshwater crayfish with a large rounded cephalothorax and rudimentary abdomen and appendages (Andrews 1904, 1907a, 1907b; Sandeman and Sandeman 1991). The fourth and fifth pereiopods have the hooked dactyli typical of parastacoid crayfish (Gurney 1942). Interestingly in embryonic Procambarus clarkii the tips of the fourth and fifth pereiopods bend back like hooks, but these are lost at hatching (Andrews 1907a). The Stage 2 juveniles of E. bispinosus are similar to those of other parastacoids (Johnson 1979; Suter 1977) and cambarids (Andrews 1907b), but differed from Stage 2 astacids in being dependent on the mother. Stage 2 astacids have well developed setae, especially the dense fringe of telson setae and an elongate cephalothorax similar to Stage 3 parastacoid juveniles (Andrews 1907b; K6ksal 1988; Lowery and Holdich 1988; Mason Proc. LINN. SOC. N.S.W., 119. 1998 J.A. HONAN 53 1977). Stage 2 Cambaridae and Parastacoidea are more rotund, have poorly developed sensory setae, and small setal buds on the telson (Andrews 1904, 1907a, 1907b; Rudolph and Rios 1986; Rudolph and Zapata 1987). The exceptions to this are Stage 2 Astacopsis gouldi and A. franklinii juveniles which have rudimentary uropods (Hamr 1992). Stage 3 E. bispinosus juveniles were independent and were similar to juveniles of other parastacoids (Johnson 1979; Suter 1977). They had similar heavy pigmentation and body sculpture to Astacopsis (Hamr 1992), reflecting the larger egg size, longer incuba- tion time and heavier body sculpture of the adults. Hatching, moulting and independence were all synchronous in E. bispinosus and in E. spinifer (Turvey and Merrick 1997); other genera are less so (Hopkins 1967; Suter 1977; Rudolph and Rios 1986; Rudolph and Zapata 1987), perhaps to avoid overcrowd- ing and improve survival of the entire brood (Hopkins 1967). The similarities and differences between the eggs, pleopods and juveniles of the astacuran superfamilies support the current phylogenetic relationships. The differences between the Astacoidea and Parastacoidea in the attachment of Stage 1 and 2 juveniles support independent invasions of fresh water by these two superfamilies (Gurney 1942). This is supported by the different egg shapes (Honan and Mitchell 1995a), hatching and telson connections. Within the Astacoidea there are significant differences between the Cambaridae and Astacidae, particularly the stage at which they become independent. Conversely the similarity of egg and juvenile development within the ecologically diverse Parastacidae indicates that these species probably had a common invasion of fresh water. An exception to this is the Tasmanian genus Astacopsis which warrants fur- ther investigation. ACKNOWLEDGEMENTS The Department of Natural Resources and Environment, Victoria (formerly the Department of Conservation, Forests and Lands) provided permits to catch crayfish. This work was part of a Master of Science degree supervised by Dr Brad Mitchell (Deakin University) and Dr Sam Lake (Monash University) at Deakin University. Thanks to David Hoey, Dr Lydia Mayner, Chris Walsh and Bob Collins for their much val- ued advice and assistance. The suggestions of the referees were gratefully incorporated in an earlier draft of this paper. This study was supported by a Commonwealth Post-graduate Research Award. REFERENCES Andrews, E.A. (1904). Breeding habits of crayfish. American Naturalist 38, 165-206. Andrews, E.A. (1907a). The attached young of the crayfish Cambarus clarkii and Cambarus diogenes. American Naturalist 41, 253-274. Andrews, E.A. (1907b). The young of the crayfishes Astacus and Cambarus. Smithsonian Contributions to Knowledge 35, 1-80. Clark, E. (1937). The life history of the Gippsland crayfish. Australian Museum Magazine 6, 186-192. Cukerzis, J.M. (1988). Astacus astacus in Europe. In “Freshwater Crayfish: Biology, Management and Exploitation”. (Eds D.M. Holdich and R.S. Lowery.) pp. 309-340. (Croom Helm: London.) Gurney, R. (1935). The mode of attachment of the young in the crayfish of the families Astacidae and Parastacidae. Annals and Magazine of Natural History 16, 553-535. Gurney, R. (1942). “Larvae of Decapod Crustacea”. (Ray Soc. Series No. 129: London.) (Reprinted 1960 as “Bibliography of the Larvae of Decapod Crustacea and Larvae of Decapod Crustacea”. H.R. Engelmann (J. Cramer) and Wheldon and Wesley, Ltd. Weinheim/Bergstr. Codicote/Herts). Hamr, P. (1992). Embryonic and postembryonic development in the Tasmanian freshwater crayfishes Astacopsis gouldi, Astacopsis franklinii and Parastacoides tasmanicus tasmanicus. Australian Journal of Marine and Freshwater Research 43, 861-878. Hessen, D.O., Taugbol, T., Fjeld, E. and Skurdal, J. (1987). Egg development and lifecycle timing in the noble crayfish (Astacus astacus). Aquaculture 64, 77-82. Holdich, D.M. and Reeve, I.D. (1988). Functional morphology and anatomy. In “Freshwater Crayfish: Biology, Management and Exploitation”. (Eds D.M. Holdich and R.S. Lowery.) pp. 11-51. (Croom Helm: London.) Proc. LINN. SOc. N.S.W., 119. 1998 54 EGG AND DEVELOPMENT OF EUASTACUS Honan, J.A. and Mitchell, B.D. (1995a). Reproduction of Euastacus bispinosus Clark (Decapoda: Parastacidae), and trends in the reproductive characteristics of freshwater crayfish. Marine and Freshwater Research 46, 485-499. Honan, J.A. and Mitchell, B.D. (1995b). Catch characteristics of the large freshwater crayfish Euastacus bispinosus Clark (Decapoda: Parastacidae), and implications for management. Freshwater Crayfish 10, 57-69. Honan, J.A. and Mitchell, B.D. (1995c). Growth of the large freshwater crayfish, Euastacus bispinosus Clark (Decapoda: Parastacidae). Freshwater Crayfish 10, 118-131. Hopkins, C.L. (1967). Breeding in the freshwater crayfish Paranephrops planifrons White. New Zealand Journal of Marine and Freshwater Research 1, 51-58. Johnson, H.T. (1979). Reproduction, development and breeding activity in the freshwater crayfish Cherax destructor Clark. M.Sc. Thesis, University of Sydney, New South Wales. Koksal, G. (1988). Astacus leptodactylus in Europe. In “Freshwater Crayfish: Biology, Management and Exploitation”. (Eds D. M. Holdich and R. S. Lowery.) pp. 365-400. (Croom Helm: London.) Lowery, R.S. and Holdich, D.M. (1988). Pacifastacus leniusculus in North America and Europe with details of the distribution of introduced and native crayfish species in Europe. In “Freshwater Crayfish: Biology, Management and Exploitation”. (Eds D.M. Holdich and R.S. Lowery.) pp. 283-308. (Croom Helm: London.) Mason, J.C. (1977). Artificial incubation of crayfish eggs (Pacifastacus leniusculus (Dana)). Freshwater Crayfish 3, 119-132. Morgan, G.J. (1986). Freshwater crayfish of the genus Euastacus Clark (Decapoda, Parastacidae) from Victoria. Memoirs of the Museum of Victoria 47, 1-57. Morgan, G.J. (1997). Freshwater crayfish of the genus Evastacus Clark (Decapoda: Parastacidae) from New South Wales, with a key to all species of the genus. Records of the Australian Museum, Supplement 23, 1-110. Morrissy, N.M. (1970). Spawning of marron, Cherax teniumanus (Smith) (Decapoda: Parastacidae) in Western Australia. Fisheries Bulletin of Western Australia 5, 1-34. Reichenbach, H. (1877). Die Embryonalanlage und erste Entwicklung des Flusskrebses. Zeitschrift fuer Wissenschaftliche Zoologie 29, 123-196. Cited in Huxley T.H. (1880). “The Crayfish: An Introduction to the Study of Zoology”. (Kegan Paul Tench, London.) Reichenbach, H. (1896). Stuien zur Entwicklungsgeschichte des Flusskrebses. Abhandlungen der Senckenbergischen Naturforschenden Gesellschaft 14, 1-138. Cited in MacBride, E.W. (1914). “Textbook of Embryology”. Vol. I. (MacMillan, London.) Rudolph, E.L. and Rios, J.O. (1987). Ontogenic development of the Chilean burrowing crayfish Parastacus pugnax (Poepigg, 1835), under laboratory conditions. Biota (Osorno, Chile) 3, 45-58. Rudolph, E.L. and Zapata, L.R. (1986). Embrional and post larval development of the Chilean burrowing cray- fish Parastacus nicoleti (Philippi, 1882), under laboratory conditions. Biota (Osorno, Chile) 2, 37-50. Sandeman, R. and Sandeman, D. (1991). Stages in the development of the embryo of the freshwater crayfish Cherax destructor. Roux’s Archive of Developmental Biology 200, 27-37. Stko, T. (1962). Studies on the development of the crayfish VII. The hatching and the hatched young. Science Reports of Saitama University Ser. B. 4, 37-42. Suter, P.J. (1977). The biology of two species of Engaeus (Decapoda : Parastacidae) in Tasmania. II. Life history and larval development with particular reference to E. cisternarius. Australian Journal of Marine and Freshwater Research 28, 85-93. Thomas, W.J. (1970). The setae of Austropotamobius pallipes (Crustacea : Astacidae). Journal of Zoology London 160, 91-142. Thomas, W.J. (1991). Aspects of egg attachment in Austropotamobius pallipes (Lereboullet, 1858) (Decapoda, Astacidae). Crustaceana 61, 287-293. Turvey, P. and Merrick, J.R. (1997). Reproductive biology of the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales 118, 131-155. Wear, R.G. (1974). Incubation in British decapod Crustacea, and the effects of temperature on the rate and suc- cess of embryonic development. Journal of the Marine Biological Association of the United Kingdom 54, 745-762. Yonge, C.M. (1937). The nature and significance of the membranes surrounding the developing eggs of Homarus vulgaris and other Decapoda. Proceedings of the Zoological Society of London 107a, 449-517. Proc. LINN. SOC. N.S.W., 119. 1998 Growth, Catch Rates and Notes on the Biology of the Gippsland Spiny Freshwater Crayfish, Euastacus kershawi (Decapoda: Parastacidae), in West Gippsland, Victoria JOHN L. MOREY (Communicated by J.R. Merrick) Flora, Fauna and Fisheries Division, Department of Natural Resources and Environment, 71 Hotham Street, Traralgon, Victoria 3844 Morey, J.L. (1998). Growth, catch rates and notes on the biology of the Gippsland Spiny Freshwater Crayfish, Euastacus kershawi (Decapoda: Parastacidae), in west Gippsland, Victoria. Proceedings of the Linnean Society of New South Wales 119, 55-69. Over ten years (1987-1996 inclusive), 11 sites in the Bunyip and Latrobe River sys- tems were monitored annually for the Gippsland Spiny Freshwater Crayfish, Euastacus kershawi. Another two sites in these catchments were sampled intermittently. During this period, 1063 crayfish were collected, with 102 individuals recaptured at least once. Growth rates calculated from recaptured crays vary widely, with a mean annual Occipital Carapace Length (OCL) increment of 6.7 mm/year. Regression analyses indicate a decreasing annual increment with increasing OCL for females, but a relatively uniform annual OCL increment for males. Males show a significant increase in annual weight increment against mean weight, whereas females exhibit only a slight increase in annual increment with increasing mean weight. Recapture data and associated observations also suggest small resident ranges (<160 m), territorial defence behaviour and dimorphism in abdominal width; mature females have broader abdomens than adult males. Females carry between 200 and over 1000 eggs or juve- niles. Individuals often emerge on to banks and burrowing activity was observed from October to mid-December and again in mid-February. For the first four years of the survey the Latrobe River system was closed to the recre- ational taking of freshwater crayfish, permitting a comparison with catch rates in subsequent years; however, lack of change in harvests or size structure of populations sampled suggest that the closure did not have a significant effect on the E. kershawi fishery. Manuscript received 6 October 1997, accepted for publication 15 January 1998. KEYWORDS: Euastacus kershawi, Gippsland Spiny Freshwater Crayfish, growth rates, catch rates, annual OCL increment, annual weight increment, fishery closure. INTRODUCTION The Gippsland Spiny Freshwater Crayfish, Euastacus kershawi (first described by Smith 1912), is one of ten species of spiny crayfish in Victoria and is indigenous to most rivers and creeks in Gippsland. Its range extends from about the Bunyip River to almost the New South Wales border in southerly flowing streams (Clark 1936; Morgan 1986). Although the genus Euastacus is well represented in eastern Australia, little information has been published on the ecology of these species. Studies by Clark (1937), Morgan (1986, 1988, 1989) and Riek (1951, 1969) included some ecological and biological observations, but data on growth and catch rates or population structure are only avail- able for two species. Honan and Mitchell (1995a, b, c) investigated another large Victorian species, E. bispinosus, while the research reported by Turvey and Merrick (1997a,b,c,d,e) focused on E. spinifer from New South Wales. Proc. LINN. Soc. N.S.W., 119. 1998 56 NOTES ON BIOLOGY AND CATCH RATES OF EUSTACUS KERSHAWI BASIN J & kilometres 72Q Traralgon re) LA TROBE RIVER Moondarra Reservoir a iS 5 g & Storage Dam Ro) 5 Hawthom of A . oojee O 5 Latrobe * Deep Ck Grea Figure |. Sites surveyed for Euastacus kershawi, in the Bunyip and Latrobe River basins. Key to symbols: ®@ = sites surveyed regularly 1987-1996; O = sites surveyed 1987 only; ll = sites surveyed intermittently since 1987. The Latrobe system was closed to the taking of crayfish 1982-1990. BUNYIP RIVER BASIN Proc. LINN. SOC. N.S.W., 119. 1998 J.L. MOREY 57 Euastacus kershawi 1s one of the larger freshwater crayfish. It is considered some- what of a delicacy amongst anglers and is actively fished for in spring and early summer. Concerns expressed by some groups over the exploitation of the species resulted in a ban on the taking of all species of freshwater crayfish in the Latrobe River system (along with several other Victorian river systems) in 1982. No prior surveys or monitoring was undertaken to justify the closure (Barker 1990; Morey 1987). Following a review of the management of freshwater crayfish throughout Victoria, (Anon 1992; Barker 1990; Morey 1987), new regulations were introduced on | January 1991 removing the ban on the taking of crayfish in waters that were previously closed. At the same time, a general size limit of 90 mm Occipital Carapace Length (OCL) was imposed, except for the Glenelg River system (100 mm OCL), with a bag limit of ten per person per day (5 for the Glenelg River system). Gear restrictions were also added to the existing Recreational Fishing Regulations for Spiny Freshwater Crayfish (CNR 1992). Objectives of this paper are: to analyse recapture data from long-term surveys to determine annual growth rates in terms of occipital carapace length (OCL) increments and annual weight increments; to document field observations on local movement, defence behaviour, reproduction and burrowing; to compare catch rates during and after a closure and gauge effects of restrictions on the E. kershawi fishery. METHODS Of the original 21 sites surveyed (Morey 1987), 11 sites in the Latrobe River catch- ment were monitored annually from 1987 to 1996, and a further two sites within the Bunyip River catchment were surveyed intermittently (Fig. 1). Sampling was conducted for four years (1987-1990) during the closure in the Latrobe River system. The other eight original sites were not monitored further because either no crayfish were collected in the 1987 survey or the sites could not be precisely relocated. Originally, sites were selected on their accessibility (close to roads and popular with anglers), or their inaccessiblity (access only through private property) as controls. Most sites were located in cleared pasture, below the 200 m contour. Riparian vegetation consisted primarily of introduced pasture species, with an over-storey of willows, (Salix spp) or remnant Eucalyptus spp, with the exception of site 6 (Tyers River) and site 11 (Traralgon Creek), which both flowed through relatively natural vegetation. At each site, baited drop nets (hoop nets) were deployed during daylight hours (0930-1730 hrs) when E. kershawi is reported to be most active. Nets were placed at 10- 20 m intervals over a distance of approximately 200 m, preferably in the calmer water associated with holes in the river bed, but to retain the 10-20 m interval some nets were placed in faster flowing waters. The nets were lifted every 30 minutes over a period of between 2.5—4 hours (5-8 lifts). Any crayfish collected was sexed, weighed and mea- sured (OCL), and individually tagged by clipping the telson and uropods as per Hume (1986). It was possible to detect tag marks for up to four years after initial tagging. Although some tag marks had healed over, scarring was evident at tagging sites on the tailfan. Specimens were returned to the water at site of capture and the nets replaced. Regression analysis was used to analyse annual incremental growth with respect to OCL and weight, for both males and females. Analysis of variance using generalised linear models in Genstat was used to compare mean catch rates between open and closed seasons. RESULTS 1063 crayfish have been caught, measured and marked at the 13 monitoring sites since 1987, but at most sites there were also escapes, individuals that fell through the Proc. LINN. Soc. N.S.W., 119. 1998 58 NOTES ON BIOLOGY AND CATCH RATES OF EUSTACUS KERSHAWI 1800 1600 y = 0.0008x7*"” R’ = 0.9758 1400 1200 1000 WEIGHT (g) 800 600 400 200 50 100 150 200 OCL (mm) Figure 2. Relationship of occipital carapace length (OCL) to weight for both sexes of E. kershawi, in the 3unyip and Latrobe River systems. n = 1,093 (includes recaptured specimens). Proc. LINN. SOc. N.S.W., 119. 1998 J.L. MOREY 59 mesh or were below or not completely in the net when lifted. The overall sex ratio at all sites was almost 1:1 (518 males, 564 females, 4 unknown or abnormal). The OCL- weight relationship indicated a non linear relationship with all crayfish gaining weight exponentially with length (Fig. 2). Annual growth rates were calculated for recaptured specimens (change in OCL over time). Although three females showed no increase at all over a twelve month peri- od, the overall mean annual growth increment was 6.7 mm/year. Individual increases varied from a minimum of 2 mm over a three year period to a maximum of 19 mm in twelve months, (three individuals — Table 3). Regression analysis for OCL increments for both sexes indicated there was a significant decrease in annual OCL increment (P<0.01), for female E. kershawi. The larger the individual, the less it grows in OCL each year (Fig. 3a). For males, there was no significant decrease in annual OCL incre- ments (P = 0.81), suggesting a relatively uniform growth rate (Fig. 3b) The two largest recaptures were males (initial OCL 130 mm, 112 mm). Their OCLs increased by 15 mm over a 2 year period and 24 mm over a 3 year period respectively (average annual incre- ments of 7.5 mm and 8 mm respectively). There were several “outliers” in Figure 3, these were rechecked and confirmed before inclusion. In one instance, two individuals from the same survey site (Bear Creek), had an annual increment of 19 mm, and one cray with a similar annual increment (19 mm) was recaptured from the Tanjil River. Linear regressions of annual weight increases to mean weights indicated that there was no significant annual increase for females (P = 0.38), whereas there was a significant (P<0.0001) increase in annual increments as the males increased in mean weight, partic- ularly at sizes exceeding 100 mm OCL and 600 g (Figs 4a,b). The placement of a log curve through the male data points appears to better represent this relationship (Fig. 4b). Tagged crayfish were often recaptured in the original vicinity of capture, some up to four years after initial tagging. Others have been re-caught at varying distances (10 m to 160 m) away from the original capture site. Although several crayfish were caught in the same location (i.e. the same net) in successive lifts, very few lifts resulted in two or more crayfish in the same net. This behaviour tends to indicate a territorial defence behaviour where crayfish will defend their territory and/or food from others. Crayfish kept in close proximity to each other will fight, often inflicting severe damage, such as a crushed or punctured carapace or the loss of limbs. Many of the crayfish caught exhibit- ed signs of injury such as lost or regenerating forelegs, tips of claws etc. (for example, 27 of 106 crayfish (25%) exhibited visible damage in the 1990 survey). In this study only 30 (5.5%) of the 546 females captured were “in berry” or car- rying juveniles, although almost 40% of females caught were considered sexually mature. Female E. kershawi had relatively broader abdomens than males and although abdominal widths were not measured, it was easy to identify mature females this way. Estimates of ova or young carried varied from 200 to 1000+ per female, the higher fig- ure being the norm. With the exception of five females from Traralgon Creek (OCLs 65 mm, 67 mm, 69 mm, 73 mm and 80 mm), all berried E. kershawi had an OCL of 87mm or more. E. kershawi were often observed on the banks adjacent to rivers and creeks, and, according to some landholders and recreational fishermen, crays have been found some- times well into adjacent paddocks many metres from the waters edge. E. kershawi were observed excavating burrows in the banks of the Tanjil River in mid December and mid February, but it is not known whether they construct type 1b (connected to the permanent water) or type 2 burrows (connected to the water table) (Horwitz and Richardson 1986) or what sex they were. Freshly excavated burrows adjacent to waters have also been observed in October and November. A total of 1086 crayfish were collected between 1987 and 1996 in a total of 6,414.5 net hours, an overall catch rate of 0.17 crayfish per net-hour for all sites sam- pled. Numbers caught at each site varied, but were generally fairly low (Table 1) with a Proc. LINN. SOC. N.S.W., 119. 1998 NOTES ON BIOLOGY AND CATCH RATES OF EUSTACUS KERSHAWI 60 ‘sayeu poinjdeoal (q) ‘sayeuay poinjdesal (8) :(9661-L 86] )SuiaIsAS aqoneyT pur diAung oy) Woy MmMoYsiay “| IO} TOO ULdUI 0} JUSWIIIOUI YSU] ddedesed [eIdI990 Jo drysuonejay “¢ 241324 (WW) TOO NVAWN O71 OIT OOI 06 08 OL 09 ]_ tt Z6v1'0 = 7a €66'€I + X8r760'0- = A : UVAA Wd (ww) LNAWHYONI TOO 119. 1998 N.S.W., Proc. LINN. Soc. 61 J.L. MOREY €100°0 = -u SOEE'6 + XEL00'0- = A (ww) TOO NVAW OOT 06 SSS et O° 08 OL 09 Ol cl vi 91 81 0c "Sa[BUl panjdedal (q) ‘panuluUod ¢ ainsi UVAA Wdd (ww) LNAWAUYONI TOO Proc. LINN. SOc. N.S.W., 119. 1998 NOTES ON BIOLOGY AND CATCH RATES OF EUSTACUS KERSHAWI “saTeu painjdeoal (q) SsaTeulay painjdvoas (8) “(966I-L861) SWaISAS agoeT] pure diAung oy) WO, IMDYsSiay “J OJ JYSIOM ULIUT 0} JUSWIDIOUI JYSIOM [enuUR Jo diysuoNLlay “fp ains1y (6) IM NVA 008 OOZ 009 00S OOv OO€ 002 OOl ccc ° 0°09 ° ° ZeLo0=,4 Wins ° 799'8S + Xpbz0'0 = A Bee OOl ° ° OcL ° OVl (6) INSWSYONI IVANNV Soc. N.S.W., 119. 1998 LINN. Proc. 63 J.L. MOREY OOrl OO0Cl OOO}! 61S7'0 = 24 ZL 8vz - (x)UTPE6 6S = A 6180 = 24 BLE ES + XETZLO =A (6) LHDISM NVAW 008 009 OOV “sayeul painjdeoal (q) ‘panunuUod p ainsi] 002 OS e) fe) (6) LNSWAYONI IWANNV Oo Ww) _- 002 OSc 119. 1998 Proc. LINN. SOC. N.S.W., 64 NOTES ON BIOLOGY AND CATCH RATES OF EUSTACUS KERSHAWI maximum of 23 individuals caught during any one day during a survey (Site 8 in 1996). Catch rates were also generally low (Table 1) with a maximum of 0.50 crayfish per net- hour during any one survey (Site 6 in 1996). Average catch rates between 1987-1990 (during the closed season) and 1991-1996 (after the closed season) increased at six of the 11 regularly surveyed sites (Sites 1, 2, 6, 8, 10 and 13), declined at 3 sites (Sites 3, 7, and 11) and remained con- stant at two sites (Sites 4 and 9). The mean catch rate at the two irregularly surveyed sites also remained constant. Analysis of variance using generalised linear models on the mean catch rates showed there was no SSMU difference between open and closed seasons (P = 0.147). Similarly, the size/catch rate ratio also did not alter significantly (Table 2). Of the crayfish collected, 102 or 9.6% were recaptures, having been tagged on previous surveys (Table 3). Only two crayfish were recaptured again on the same day they were tagged. Catch rates were related to the time that nets were left set, declining markedly the longer the nets were left immersed. When baited nets were left for a minimum of 2.5 hrs soak time, i.e. five lifts, 70% of the total catch was caught within the first three lifts (Table 4). Nets left longer than 2.5 hrs (up to a maximum of 4 hrs) still caught crays, but catches were lower (5—10 individuals on lift 7 or 8). DISCUSSION E. kershawi individuals grow very slowly. If E. kershawi grow at a uniform 6.7 mm/year (the average annual growth increment), a crayfish of minimum legal size of 90 mm OCL is about 13 years old. This is in close correlation, but a slightly slower growth rate, from estimates of size and age for E. bispinosus, as proposed by Honan and Mitchell (1995b) who estimated E. bispinosus to have lifespans exceeding 25 years. Even at a uniform rate of the highest growth rate recorded of 19 mm per year, a Gippsland Spiny Crayfish of minimum legal length is about 5 years old. The largest cray collected during this study had an OCL of 151 mm (from the Latrobe River in 1990), however a male specimen collected by a recreational angier in December 1996, also from the Latrobe River, at Noojee, had a confirmed OCL of 165 mm, 5.5 mm larger than the maximum OCL reported by Morgan (1986). OCL increments were not uniform and varied between individuals and also years. For example, one individual grew 10 mm one year and 3 mm the following year. However, average growth rates at sites where a reasonable sample of recaptures were found (>5) was fairly constant (6.7—-10.6 mm/year). Two of the three individuals that recorded the maximum increment of 19 mm were from the same site. There is no obvi- ous explanation for these high values, but it is possible that these individuals had com- pleted a double moult (i.e. they had moulted twice within the twelve month period). Honan and Mitchell (1995b) estimated that E. bispinosus individuals less that 45 mm OCL moulted twice annually and once per year for individuals over 55 mm OCL, although some large specimens (>110 mm OCL) did not moult at all or had a small increment. Similarly, Turvey and Merrick (1997d), estimated E. spinifer moults three times annually for individuals <35 mm OCL, twice per annum for individuals up to 55 mm OCL and annually for specimens over 55 mm OCL. With the exception of one indi- vidual (with initial OCL of 53 mm), all recaptured E. kershawi specimens had an initial OCL of 55 mm or greater, and are estimated to have moulted only once per year, except for those individuals that had either exhibited no annual increment, or had increased at approximately three times the annual average increment. Male E. kershawi with OCLs in excess of 100 mm continued to grow at the annual average increment, whereas females showed a distinct decline in annual OCL increment, particular over 80 mm OCL. This is in contrast to Turvey and Merrick (1997e), who Proc. LINN. SOC. N.S.W., 119. 1998 a) J.L. MOREY ‘AOAINS SABP UO UY) JIOW JIAO JYBNLS SIOqUINA, x. (COVOE'P (pO'O)Z (11O)S (€1'0)9 (IOP = = (OL'OE (€1'0)8 i = (LO'O)b 61 (VOT b —(ZO'O) «I (80'O)P (prOoL (61'OL —— (110) x2 (IV Op (80'O)x€ = = (plOE LI (TOES (COLO)LI (LI'0)8 (IOP (LO'OE (Or O)rl (LV'O)O1 (OC OE TI (L7'0)6 (9¢'0)91 (ZE'O)SI (17'0)6 el (IV Oe's (€1'0)9 = (90'O)E (S1'0)8 = (Or (61 OL (ET OPI (IOP (L70)9 (S1'O)r a (61'0)8'8 (90'O)€ (81°06 (pr'0)07 (STOL = (€L'O)S (p1'0)8'P (Z1'0)9 (17'0)9 (07'0)9 (pO'O)I Ol (TOTTI —(COZ'OOI (ZE'O)SI (1e'O)rI (L1'0)8 = (60°O) «PI (ZTZO)B'EL (Z1'0)%8Z (87'0)6 (91'0)9 (€€O)TI 6 (CLOSIZ (8Pr'O)ET (9¢'O)8I (9b' 0) (CLOT (87 O)ET (970) «17 (87O)SEL (ZE'O)9I (ve'0)6 (ZE'0)07 (y1'0)6 8 (ZOE SI (STOL (pr'O)ZZ (re OLI (60'O)ZI (OVOEL (LOO)«1Z (pTOEOT (ELOxIr (OPrOsl (€7'0)9I (81°0)9 [l (SeOEI (0S O)6I (€COTI (b7'O)8 (Sr'ODLI = (€7'0)6 (11'O)8"€ (61 OL (90'0)Z (ZOO) (ST'O)S 9 (rOaL (11O)S (90'O)E (pr OL (81'0)8 = (SO'O)x€I (11'0)9 (60'0)x€1 (ST OL (€0'O)I (SO'OE i (O1'0)9°r (60'O)P (ST'OL (OV'0)S (60'O)r = (80'O)E (STOE'S (80'O)r (87'0)8 (91'0)9 (80'OE € (e081 (61'0)6 (LEO)rl (pr'O)8I (eo (0€'0)6 (SZ O)0I (87TOSIT (87 OLI (S1'O)S (Ir OZI (97'O)EI (4 COST (Se Ol (6€'O)SI (LTO)TI (ZZ'0)0I (0€'0)6 (S7'O)0I (€1'0)8 (€1'0)8 (07'0)8 (90'0)S (ZVO)I I uodo posoyo uvolul 9661 661 661 £661 7661 1661 urow 0661 6861 8861 L861 aus ‘766 UL SADAINS DWIOS PaUdADId JOYVOM JUDWIOJOUT “(IID UINQoTIUUTY pue JOATY OSVIL]) 6 PUL LT SOUS Jdaox9 si9IeM [JP UL [66] [HUN YSIJALID JO SULYL) JY) UO UOSvaS Paso]d B SEM JOY L, ‘Sosa Used ur (ANOY-JoU Jad YSTJARAD) 991 YOIVD YIM OPIS YOwd ye IIA Yow UT IYSNd IMDYSAaY SNODISONY JO SIOQUINN | dav, Proc. LINN. SOc. N.S.W., 119. 1998 NOTES ON BIOLOGY AND CATCH RATES OF EUSTACUS KERSHAWI 66 (%88)L'€ (%001)T (%r1)9'0 (STVTI (PDL (%91)r'I (%9V)T (%1)8°T (DLT (%e)v'0 (Ste (%€)8'1 (OC (Bp)TS uPvOoUT ‘7661 Ul POADAINS JOM SOUS [[B ION “YoIed [eI0} Jo (Sasayjuared ul) advyUadIed puv YYSuIT dovdvivg [eUdi99Q IIOUW JO WLS MMYYSMay SNOVISYN JO JOQuInN (%0)0 (%9)1 (%0)0 (%0)0 (%09)9 (%6)T (%0)0 (%S)1 (®OWE (%ST)I (bby (%T)8 9661 (%001)S (SSO) (SOT (%1)I (%LU)v (%0)0 (%S)1 (%0)0 (%EE)I (%EV)E (%60)v (%OTE S661 (%0)0 (%0)0 (%L9)T (%SDE (%0)0 (%7O)L (%9)I (SEDI (%rI)I (%0)0 (%bv)8 (%L9)8 uado y66l (%E8)S (BSLIE (%6V)T (%0)0 (%€1)1 (%6T)T (%0)0 (BLUE (EOP (%0)0 (%E9)S (SLE (LOE (%0b)P c66l (%P VT (%8)1 (%ETIE (%0)0 (%EE)E c661 (%0)0 (%0Z)C (%0)0 (%07)1 (%0)0 (6D) (%S)I (%0)0 (%8E)S (%99)C (%09)9 (%0S)S 1661 ZT ATV; (%€E8)S (%0)0 (%Ip)s (4D)L'0 (%E)L'I (%p)S'S (OEP (%T)8'P (%0)0 (%EE)T (4p)ET (M)8'P (%S)8'P uevouwl (%SL)9 (%0)0 (BLEVL (%L)I (BEET (YEv TI (YE (%T1)S (%0)0 (%9b)9 (MSLIE (%69)L (%€9)S 0661 (%8)9 (SST) (%EE)T (BECOVE (%0)0 (%€€)9 (%0)0 (%r DI (%EDI (%OPIT %(E9)S pasoya 6861 (%OTE (LD) (%0S)E (SLDI (%09)01 (%1E)S (%0)0 (%0)0 (ECT (%07)1 S8ol (%0)0 (Mpb)b (%0)0 (%0)0 (%09)9 (%rp)P (WOSE (%0)0 (%EE)] (BODE (%EL)8 L86l (%00D)r nm los | ALIS Proc. LINN. SOC. N.S.W., 119. 1998 J.L. MOREY 67 reported no difference in growth related to sex for E. spinifer. France (1985) noted annu- al growth rates declined with age for a North American freshwater crayfish, Orconectes virilis. Similarly, Turvey and Merrick (1997e) and Honan and Mitchell (1995b) reported declining annual increments for E. spinifer over 55 mm OCL, and E. bispinosus over 110 mm, respectively. Why male E. kershawi appear to continue growing at a relatively con- stant rate is unexplained. The largest female E. kershawi captured had an OCL of 122 mm, whilst there were 10 males with an OCL of 122 mm or more. The low recovery rates in this study may indicate either heavy fishing pressure, mobile populations or high population numbers within these waters. The catch/effort results and anecdotal evidence tend to suggest fairly low resident populations due to fish- ing pressure. Honan and Mitchell in their study on catch characteristics of E. bispinosus (1995c) found most tagged individuals moved less than 75 m from the capture site. Similarly, most recaptured E. kershawi (85%) were recovered within 60 m of the original capture site. Honan and Mitchell (1995a) also noted some E. bispinosus exhibited miss- ing appendages or wounds, and in their 1995b report mentioned one in four large E. bispinosus with missing or regenerate chelae. Similar observations have been reported by Bicskos (1989), for E. armatus. Damage to individuals is consistent with territorial defence behaviour. All five berried females from Traralgon Creek had an OCL of 80 mm or less, whilst berried E. kershawi from all other sites had an OCL of 87 mm or more. 82.6% (19 females) had an OCL of 90 mm or more, but it is not known why the Traralgon Creek population matures at a lesser OCL than all other sites. Six E. yarraensis, a sympatric species in the Tarago River, were collected during the survey period, including one female in berry with an OCL of 52 mm. E. yarraensis were included in the catch rate data, however no recaptures of previously tagged speci- mens precluded any growth estimates. TABLE 3 Recapture and growth rates of Euastacus kershawi, for the 11 regularly and 2 irregularly surveyed sites. Two “outliers” (due to recording errors), not included. Location (Site) Total Caught Recaptures Percentage of OCL increase Average annual recaptures range (mm) OCL increase (mm) Tanjil River (1) 100 6 6.0 1.7-10 5.5 (2) 101 15 14.9 2-19 7.4 Latrobe River (3) 44 0 0 — — (4) 54 5) 93 5-8 ell Tyers River (6) 76 4 Ss) 6-9.3 7.6 Shady Creek (7) 166 22 13.3 0-18 8.0 (8) 130 23 Nod 0-10 6.9 Narracan Creek (9) 108 10 9.3 5.5-15 8.7 (10) 60 3 5 6-10 8.3 Traralgon Creek (11) 46 2 4.3 1-3 1.8 Bear Creek (13) 109 7 6.4 0-19 VS Tarago River (17) 44 1 2.3 0 0 Minnieburn Creek (19) 25 4 16 0.7-8 4.4 TOTAL 1063 102 9.6 0-19 6.7 Proc. LINN. Soc. N.S.W., 119. 1998 68 NOTES ON BIOLOGY AND CATCH RATES OF EUSTACUS KERSHAWI TABLE 4 Catch of Euastacus kershawi compared to lift, when nets were left for a minimum of 2.5 hours, (nets were lifted every 30 minutes, 1.e. five lifts), for all surveys 1990 and 1991. Lift No. 1 2 3 4 5 No. of crayfish 72 52 35 32 33 Cumulative total 72 124 159 19] 224 Percentage of total 32% 55% 71% 85% 100% In contrast to this study, where low numbers of berried E. kershawi were recov- ered, Honan and Mitchell (1995a) reported up to 95% of mature female E. bispinosus carried eggs in the breeding season and remained active and catchable. Like some other freshwater cray species, it is possible that berried E. kershawi are more secretive, trap shy, or confined to burrows, making them more difficult to catch (Abrahamson 1971; Huner 1988; Lowery 1988); have low spawning rates (France 1985; Huner and Lindqvist 1986) or that the breeding period for E. kershawi 1s outside the survey period (late August-early November), although Clark (1937) reported the breeding season for the Gippsland Spiny Crayfish as early spring. Hamr and Richardson (1994) reported Parastacoides tasmanicus and other species as having a biennial breeding cycle, howev- er Honan and Mitchell (1995a) and Turvey and Merrick(1997a) reported E. bispinosus and E. spinifer as having annual breeding cycles. What stimulates E. kershawi to leave the water and travel overland or why and when they excavate burrows is unknown and requires further research. The apparent lack of change in the catch rate and size components of the popula- tions questions the effectiveness of the closure on the taking of crayfish in the eighties. As this project progressed, it became apparent (through evidence of discarded bait lines or advice from landholders) that all the “inaccessible” sites were frequented by anglers seek- ing freshwater crayfish. Evidence of poaching at several sites (and at other locations on the Latrobe River and its tributaries) during the fishing ban, and comments from adjacent landholders also supports the apparent lack of effectiveness of the closure. Similarly, ille- gal fishing was observed in a reserve established for the the large Tasmanian freshwater crayfish, Astacopsis gouldi; the closure of the area was reported to be ineffective in limit- ing harvesting of that species (Horwitz and Hamr 1988). Non compliance with the closure of certain waters to the taking of freshwater crayfish in the eighties would also explain the similarity in the catch rates of the “open” and “closed” waters in the initial 1987 survey. ACKNOWLEDGEMENTS The assistance of R. Hasthorpe, Fisheries and Wildlife Officer, Traralgon, in the initial year of the moni- toring program is greatly appreciated. The help and enthusiasm of work experience students Jamie Magyar and Graeme Lockhead in 1991 is also acknowledged. Thanks to Greg Hollis and Paul Dignon, Department of Natural Resources and Environment (DNRE), for their valuable assistance with data analysis and to Tarmo Raadik, John Barker, Brian Ward and Tim Doeg of DNRE for their comments on earlier drafts. REFERENCES Abrahamsson, S.A. (1971). Density, growth and reproduction in populations of Astacus astacus and Pacifastacus leniusculus in an isolated pond. Oikos 22, 373-388. Anon. (1992). Fisheries Notes No 22 Victorian Department of Conservation and Environment, Melbourne. Barker, J. (1990). Spiny freshwater crayfish management strategy. Fisheries Management Report (34), 1-17. Department of Conservation and Environment, Melbourne. Proc. LINN. SOC. N.S.W., 119. 1998 J.L. MOREY 69 Bicskos, A. (1989). Sharp encounters with the Murray Cray. Geo 11, 41-47. Clark, E. (1936). The freshwater and land crayfishes of Australia. Memoirs of the National Museum Melbourne 10, 5-58. Clark, E. (1937). The life history of the Gippsland crayfish. Australian Museum Magazine 6, 186-192. CNR (1992). Victorian Fisheries (Recreational) Regulations 1992. Government of Victoria, Melbourne. France, R.L. (1985). Relationship of crayfish (Orconectes virilis) growth to population abundance and system productivity in small oligotrophic lakes in the experimental lakes area, North Western Ontario. Canadian Journal of Fisheries and Aquatic Sciences 42, 1096-1102. Hamnr, P. and Richardson, A. (1994). Life history of Parastacoides tasmanicus tasmanicus Clark, a burrowing freshwater crayfish from south-western Tasmania. Australian Journal of Marine and Freshwater Research 45, 455-470. Honan, J. A. and Mitchell, B. D. (1995a). Reproduction of Euastacus bispinosus Clark (Decapoda: Parastacidae), and trends in the reproductive characteristics of freshwater crayfish. Australian Journal of Marine and Freshwater Research 46, 489-499 Honan, J. A. and Mitchell, B. D. (1995b). Growth of the large freshwater crayfish Euastacus bispinosus Clark (Decapoda: Parastacidae). Freshwater Crayfish 10, 118-131. Honan, J. A. and Mitchell, B. D. (1995c). Catch characteristics of the large freshwater crayfish, Euastacus bispinosus Clark (Decapoda: Parastacidae), and implications for management. Freshwater Crayfish 10, 57-69. Horwitz, P. and Hamr, P. (1988). An assessment of the Caroline Creek freshwater crayfish reserve in northern Tasmania. Papers and Proceedings of The Royal Society of Tasmania 122, 69-72. Horwitz, P. H. J. and Richardson, A. M. M. (1986). An ecological classification of the burrows of Australian crayfish. Australian Journal of Marine and Freshwater Research 37, 237-242. Hume, D. (1986). Results of freshwater crayfish surveys. Freshwater Fish. Management Branch, Department of Conservation, Forests and Lands, File 42/14/7. (unpublished). Huner, J.V. (1988). Procambarus in North America and elsewhere. In ‘Freshwater crayfish: biology, manage- ment and exploitation’. (Eds D.M. Holdich and R.S. Lowery) pp 239-261. (Croom Helm Ltd., London). Huner, J.V. and Lindqvist, O.V. (1986). A stunted crayfish Astacus astacus population in central Finland. Freshwater Crayfish 6, 156-165. Lowery, R.S. (1988). Growth moulting and reproduction. In ‘Freshwater crayfish: biology, management and exploitation’. (Eds D.M. Holdich and R.S. Lowery) pp. 83-113. (Croom Helm Ltd., London). Morey, J. L. (1987). Freshwater crayfish survey, Latrobe River system. Department of Conservation Forests and Lands. (unpublished). Morgan, G. J. (1986). Freshwater crayfish of the genus Euastacus Clark (Decapoda : Parastacidae) from Victoria. Memoirs of the Museum of Victoria. 47, 1-57. Morgan, G. J. (1988). Freshwater crayfish of the genus Euastacus Clark (Decapoda, Parastacidae) from Queensland. Memoirs of the Museum of Victoria 49, 1-49. Morgan, G. J. (1989). Two new species of the freshwater crayfish Euastacus Clark (Decapoda: Parastacidae) from isolated high country of Queensland. Memoirs of the Queensland Museum 27, 555-562. Riek, E. F. (1951). The freshwater crayfish (Family Parastacidae) of Queensland. Records of the Australian Museum 22, 368-388. Riek, E. F. (1969). The Australian freshwater crayfish (Crustacea: Decapoda: Parastacidae), with descriptions of new species. Australian Journal of Zoology 17, 855-918. Smith, G. (1912). The freshwater crayfishes of Australia. Proceedings of the Zoological Society of London 1912, 144-171. Turvey, P. and Merrick, J.R. (1997a). Reproductive biology of the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales 118, 131-155. Turvey, P. and Merrick, J.R. (1997b). Population structure of the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales 118, 157-174. Turvey, P. and Merrick, J.R. (1997c). Diet and feeding in the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales 118, 175-185. Turvey, P. and Merrick, J.R. (1997d). Moult increments and frequency in the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales 118, 187-204. Turvey, P. and Merrick, J.R. (1997e). Growth with age in the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales 118, 205-215. Proc. LINN. Soc. N.S.W., 119. 1998 iene ia “2 “Inepaniosat ots 4 fe a Roricot Re anaes = G gpakeiesed ey heancal % Hse SS ope Aid ies at inte VEG oP m3 sronetenene rt sisabra stil ; Har Salt we msn thy Pinay ety eftsindt adi te ces ; 2 g Pian oo rey ca iD ate wales nes ptialagany 2 ve oy wns ; * Seah a ®, Aah Gz. ied anette a ere ie wie ye Ceca irl pee qr its oT ange 3 we Magyar ban, asia 25 a dk a nie? ; wi abithis ait ay Reith sah) Ps Bi me. ‘ ae he ry = _ sdiangtiyi GL RY igi Seah 5 hicteksg beatae ait ‘ a if if | a be fy) Pe } okay . 3B i is We db dnhivine 3:3 ia RA aia Saves il hs ‘ARDEP ih. AIMEE AOR tk oie [ é ee vi nie ee azeutey AYE ene ; “as rALYI sr ahivw Tok en WIL wrie: a) “rit ty, road ale daidts 184 nT, tat GEIS: ite pat : : a He Hy pois ‘other & i avr eee a aT fenlacasesaa fr ‘Papal ia SCRA ser abi aise y "2 Bird sma? oy be meee. 7 ie Uap r oe Pare: Adler: aabaib/snarit weft ry vat be te aie = ena i oa ri igtilicnooy 4 Mlialap fh. “ : a A ee ie fil eae fasted « vbr, iad PA, hAsvgds, da siioeetl re taal a er 1 ; dl - ' oie 1 198 T aay Pear ay ae eel Oh i ; Urnw Creek 4 Hydrological Sampling Site -© Falls e Growth Study Pool HM Open Cut Goldmine Spotlight Transect and Treatment Plant ___. State Forest Boundary ir { Control Catchment —-- Forest Road Lomi | Treatment Catchment Rainforest Paired-catchment Figure 2. Features of the paired-catchment sites with details of abundance assessment transects (inserts). Note that the pools drawn within each of the streams (inserts) were connected by riffles. Proc. LINN. SOC. N.S.W., 119. 1998 G.C. SMITH, A. BORSBOOM, R. LLOYD, N. LEES AND J. KEHL 75 Growth Measurements Three locations were selected as collecting sites for growth investigations; these con- sisted of two pools separated by a riffle zone in North Booloumba Creek and a single pool in Bundaroo Creek (Fig. 2). Surveys indicated that E. hystricosus would be difficult to catch regularly without significant disturbance to their refuges under rocks or logs, so blocks of tubing were deployed in each pool to facilitate capture of crays; these artificial refuges were readily utilised by juvenile E. hystricosus. Each ‘tube trap’ consisted of two to four 300 mm lengths of black plastic tubing, 19-50 mm in diameter and set into concrete; tubes were open at both ends. Sixty blocks of tube traps were deployed in the paired catchment study pools at depths of 0.15—0.45 m. Sampling occurred in two periods: 1984-88 and 1991-94. For a year following their release a target of 30 juveniles was sought for each monthly sample. Individuals were captured by carefully raising a tube trap horizontally off the creek bed and, while it was still underwater, tipping the contents into a hand held dip net. Occasionally crayfish were caught under a tube trap. Using Vernier calipers the ocular carapace lengths (OCL) of all specimens were measured (to nearest 0.1 mm) to assess the growth rate of seven cohorts from 1984 to 1994. Abundance Measures Spotlight survey transects were established on both Bundaroo and North Booloumba Creeks (Fig. 2). Numbers of juveniles seen along each transect were record- ed monthly to provide estimates of relative abundances of each annual cohort. A census involved a single observer using a 55 W spotlight powered by a 12 V battery, walking the length of each fixed transect on two successive nights. Censuses were done in the first half of the night and only carried out when creeks were at basal flow. No stones or rocks were disturbed during a census, and numbers and estimated ages of individuals observed were recorded. Surveys were carried out between 1983 and 1987. The total pool areas surveyed on each transect were 359 m* and 598 m’ for Bundaroo and North Booloumba Creeks respectively. Depths of pools ranged from 0.09-1.05 m. Abundance estimates were obtained from the larger number of recorded crays located on each transect, over the two successive nights of spotlighting. Density estimates were calcu- lated by dividing abundance numbers by the total pool area of the transect. Analyses The paired catchment study followed a “classic” BACI design (Bernstein and Zalinski 1983), where no replication of catchments occurred (Stewart-Oaten et al. 1986). Under these circumstances it is difficult to ascertain baseline data, as only relative com- parisons can be made. For control and treatment catchments the means of monthly OCL length samples for each first year cohort were divided into three groups (December—May, corresponding to the first summer and autumn as free-living individuals; June—September, the first win- ter as free-living individuals; and October-December, the post-winter period as free-liv- ing individuals prior to the release of a new cohort) and a linear regression line fitted to each grouping. These were tested for significance. Where significance occurred the slopes were tested for differences between catchments and between years. Mean OCL measurements recorded in May and December were also compared between catchments, for each first year cohort. Correlation and t-tests (Sokal and Rolf 1981) were used to test for relationships between rainfall and turbidity and to test for differences in OCL measurements between catchments only in May of each year, following the first period of growth of the free-liv- ing juveniles. Comparisons were also carried out on OCL data for October-December between the two catchments where differences were apparent. Proc. LINN. Soc. N.S.W., 119. 1998 76 JUVENILE EUASTACUS HYSTRICOSUS (uuu) pesuley 29q AON ~O 1984 \das Bny | Aine eunr Kew judy yosey| | ged uer 20 + 15 + 10 + 5 + “9) Xe) rTM ON 320 280 24 20g | | 6 1985 ey oop Tee) (Ge) fo) 1) SIS (=) We) (oe) 0) te) te) fe) fe Sf) © LORS: ae Toy Spey Gel NY SS = (widd) Ajipiquny (uidd) Ayipiquny Figure 3. Summary of monthly turbidity data for both catchments, over a six year period, related to monthly rainfall records (from Kenilworth Forestry Station). Key to symbols on opposite page. Proc. LINN. SOC. N.S.W., 119. 1998 G.C. SMITH, A. BORSBOOM, R. LLOYD, N. LEES AND J. KEHL 77 (uuu) ypeyurey oO (o) i=) N Oo FF (op) N N it i eS Cy © (Ss) €C) Qo Oo © (No = Aree Sot | 9) Rp Fo) | (widd) AjIpiquny (uuu) |pejuIeY + 320 2eq AON Po 1987 das Bny Aine eunr Kew; judy yoey| qe4 uer A (uidd) Ajipiquny Figure 3 continued. Key to symbols: [J = Booloumba Creek catchment; (_] = Bundaroo Creek catchment; + = total monthly rainfall. Proc. LINN. Soc. N.S.W., 119. 1998 78 JUVENILE EUASTACUS HYSTRICOSUS RESULTS Habitat, Environment and Water Quality E. hystricosus was found to be essentially restricted to aquatic habitats. Adults con- struct burrows in the banks of streams and under large rocks with entrances either below or above the basal water line. Juveniles occurred mainly in shallow pools, taking refuge under rocks or logs and in rock crevices. Although juveniles did dig shallow excavations under rocks, burrow systems were not observed. Juvenile E. hystricosus have been recorded in deep pools as well as runs, glides and riffles; however, preliminary surveys have suggested that they are not evenly distributed among these stream environments (Borsboom and Kehl, unpublished data). The physico-chemical properties of water flowing in the North Booloumba Creek are presented in Table 1. While there may be minor differences between catchments, this table seeks to highlight the range of conditions experienced by the crayfish living within the North Booloumba stream. Turbidity data for each of the streams is presented in Figure 3. Natural fluctuations in turbidity were as large as any fluctuations that may have been induced by mining activities in general. No relationship was observed between turbidity and monthly rain- fall data collected from Kenilworth Forestry Station (Pearson’s Product-Moment Correlation Coefficient = 0.12, df = 64, p >0.05 for Booloumba; Pearson’s Product- Moment Correlation Coefficient = 0.20, df = 64, p >0.05 for Bundaroo). TABLE | Physico-chemical water parameters measured from locations above and below the mine site on North Booloumba Creek. Data supplied by Australian Laboratory Services Pty Ltd (to Department of Minerals and Energy, Queensland) from samples submitted and analysed 2 August 1996 to 22 May 1997. Category of Parameter Parameter Values and Units Physical pH 6.5-7.4 Conductivity 142-179 pS/cm Suspended Solids 1-3 mg/L Major Cations Ca 7-9 mg/L Mg 6-8 mg/L Na 11-17 mg/L K 0.05). A comparison (student’s t-test ) of mean OCLs in May between catchments for each first year cohort indicated that mean OCLs were significantly greater in Bundaroo com- pared to North Booloumba in 1987 (t = 2.13, df = 64, p <0.025) and in 1994 (t = 2.8, df = 69, p <0.005). Other differences in mean OCLs evident in Figure 4 (particularly October 1993, 1994; November 1985; and December 1987, 1988) were investigated for statistically significant differences. Significant differences were found between the catch- ments in October 1993 (t = 4.5, df = 46, p <0.05) and October 1994 (t = 2.5, df = 63, p <0.05). Proc. LINN. Soc. N.S.W., 119. 1998 80 JUVENILE EUASTACUS HYSTRICOSUS N (oe) [ ] = Station 1 = Station 2 [_] = Station 3 fl] = Station 4 MB = Station 5 aN 00 =< i =< N BSS Oo) Turbidity (ppm) S) 8 6 4 D2 é 0 + :; 1983 1984 1985 1987 1988 Year Figure 4. Mean turbidity data for each of five hydrology stations for six years in the Bundaroo and North Booloumba catchments. Turbidity measures reflected no extraordinarily high occurrence of sediment loads in 1986 or 1987 (except at one station on Bundaroo, where the measure was not signifi- cantly different from that experienced in North Booloumba Creek); comparable measures were not available in 1994. It is difficult therefore to relate either of these significant results to turbidity. Rainfall measures also do not suggest any clear pattern as rainfall in 1986 and 1987 was fairly normal. Significant wet periods were recorded in 1983, 1988, 1989, in late 1991/92 and in 1996; but rainfall in 1993 (904 mm) and 1994 (870 mm) was well below the annual average of 1247 mm. The only relationship apparent was that changes in growth rate (and the size of individual crays) in each of the catchments was associated with the commencement of mining (1987), although this activity has not been correlated with turbidity changes. Abundance Observed numbers and yearly trends in these numbers (for both transects) are shown in Figure 6. Monthly counts record the first appearance of each age class of juve- nile crayfish and serve initially as an indication of the level of juvenile recruitment, while later in the year as an indication of juvenile survival. Observed numbers peak annually in the January—February period, then drop rapidly through winter, but recover partly in spring and early summer before commencing to drop again. Clearly these fluctuations are partially related to the detectability of crays, which is less in winter owing to their reduced activity. The abundance estimates (Fig. 6) showed that juvenile E. hystricosus had similar annual fluctuations in numbers from year to year and across catchments. Only in 1984 were the numbers released in the two catchments considerably lower than in 1985 and 1986. Proc. LINN. SOC. N.S.W., 119. 1998 G.C. SMITH, A. BORSBOOM, R. LLOYD, N. LEES AND J. KEHL 81 When standardised (by area of pools in each transect) to provide a density index, the numbers of crays per unit pool area were slightly lower to approximately the same in Bundaroo Creek compared to North Booloumba Creek. The commencement of logging in the North Booloumba catchment in May 1986 had no observable effect on early sum- mer crayfish numbers. DISCUSSION The biology of the giant spiny crayfish is in many respects similar to that record- ed for the freshwater crayfish E. spinifer for which there has been a considerable body of information recently documented (Merrick 1995, 1997; Turvey and Merrick 1997a, b, c, d, e). The timing of life history events may be slightly different. Spawning by E. hystricosus occurs in autumn (March—April) (Kehl, unpublished data) compared with early winter for E. spinifer, and eggs are carried by E. hystricosus across the winter period (egg incubation for E. spinifer is 110-140 days) (Merrick 1997). The eggs of E. hystricosus hatch from October to November (Kehl, unpublished data) compared with spring-early summer for E. spinifer. Juveniles are released late November to mid- December in E. hystricosus and early summer (Merrick 1997) for E. spinifer from the Sydney basin region. This study has been a test case for using E. hystricosus as an “indicator” of stream integrity. The initial paired comparison of logged versus unlogged catchments was blurred by the gold mining in the control catchment in 1987. This activity was beyond the control of this study and caused an increase in turbidity at one hydrology station in that year. Relationships between physico-chemical parameters investigated and E. hystri- cosus are discussed further below. Growth of freshwater crayfish is a function of both the inter-moult period and the moult increment. E. spinifer juveniles have been known to moult up to six times in their first year, although the average is 3 times per year (Turvey and Merrick 1997d). Like E. spinifer, E. hystricosus in their first year of independent life exhibit two peri- ods of growth; these occur in the first summer and autumn of their free-living exis- tence (December—May for E. hystricosus) and in the spring-summer period (October-December) following their first winter. Both E. spinifer and E. hystricosus show insignificant growth during the winter (Merrick 1997; Turvey and Merrick 1997d). This may be due to colder temperatures directly suppressing activity and therefore feeding behaviour or food availability itself. Although the OCLs of crayfish found in Bundaroo Creek were typically larger in most months, significant differences were only recorded in May 1987 and May 1994. Growth rates were however, essentially the same; i.e. slopes did not differ significantly. There was no evidence to suggest that an earlier release occurred within Bundaroo Creek in 1987 and 1994. Growth itself however may be density dependent and it is important to recognize that good survival and highly fecund adults may produce an abundance which might suppress overall growth rates of individuals. Therefore, population size was moni- tored in parallel with growth rates. Before discussing the relative abundances the following points need to be consid- ered. Censuses were only carried out at times of basal flow, as raised levels increased tur- bidity and reduced detection rates; and counts were not performed during rainy periods, as rain on the surface also impaired visibility. Whether activity levels of juveniles during low flows in dry periods, on which estimates are based, are representative of movement at other times remains open to debate. Release numbers of juveniles and subsequent abun- dance estimates give relative indications of release time of juveniles, overall fecundity and survivorship. There was a clear similarity in the timing of release of juveniles and in the subsequent pattern of decline in numbers post-release observed in both catchments. Proc. LINN. SOC. N.S.W., 119. 1998 82 JUVENILE EUASTACUS HYSTRICOSUS | Ls | 99q = 1 (Ze) | 99q AON © ze & | ES AON fo?) po il v aL | 20 | oO des a | 2 W des | — Bny iny | Ine + | inp ae {| alt unr Kew; Ke idy dL WN oF idy @ Jew | Jey | Ss) | cll IL | teu tidad ea | Ale | | | | 9aq eee Soe ae aad Neg | -_—-++—+ +++ ++ +++ +++ agngewxeraonvnorno Seti NG S24 Ossie NO OS as NES. ~ (wi) 790 ues > (wi) 790 ues 92d HEH HoH 90q | AON ne + bes Ak N lie 2 pO | das das | Bony Bny PIP | ie | | + pee eel unp | Kew | ali | idy | idy Ei i JEN re | + Eset | qe4 pi OP ee eee ver uer NODOtTNODOYTNO rae ie iD : aes! (ww) 790 ueeW (ww) 190 ueeW | e o | Figure 5. Summary of mean OCL data related to month for juvenile cohorts of E. hystricosus over a ten year period. Key to symbols: : = Booloumba Creek samples; $ = Bundaroo Creek samples Proc. LINN. SOC. N.S.W., 119. 1998 83 G.C. SMITH, A. BORSBOOM, R. LLOYD, N. LEES AND J. KEHL €661-2661 > Se Se ee). Cc cS (= Ge sok 5 «aro y ao oe (Oa 8 {~$——_— +0 c (ww) 190 ues 90q V66L-E661 ges | ue (ww) 790 ueeW (ww) 790 ueaW Figure 5 continued. 119. 1998 Proc. LINN. SOC. N.S.W., 84 JUVENILE EUASTACUS HYSTRICOSUS (2) 499 Bundaroo Creek | oneits 4 a ee oe - 82/83 © 300 FO esse - 83/84 : : ~~~ - 84/85 5 | —-— - 85/86 E | if R6/8 7 Y 200 2 5 | O re) S 100 0 (b) 400 North Booloumba Creek Cohorts hina - 82/83 Teg i OWL ie il a - 83/84 : == -.84/85 : —-— - 85/86 3 | —— - 86/87 © 200 © O ro) S 100 Figure 6. Summary of abundance estimates of five cohorts of juvenile E. hystricosus in the paired catchments; values are unstandardised for the length of survey transects. (a) Bundaroo Creek; (b) North Booloumba Creek. Proc. LINN. SOC. N.S.W., 119. 1998 G.C. SMITH, A. BORSBOOM, R. LLOYD, N. LEES AND J. KEHL 85 Furthermore, the number of juveniles released in 1984—85 and 1985-86 were clearly much higher than the numbers released in 1983-84 and 1986-87; a pattern which was also consistent between catchments. While 1983 had been a wetter than average year, there is no suggestion that this was the primary cause of the differences noted. The effects of differing densities between years did not produce effects upon growth, although differ- ent densities between streams may in part explain why individuals were almost always a little larger in Bundaroo compared with North Booloumba. Clearly the observed differ- ences cannot be easily explained on the basis of density-dependent effects. Similarly, attempting to relate differences in growth to disparate physico-chemical parameters is also difficult. While turbidity measures within the streams did not differ appreciably between catchments, turbidity was typically higher in the North Booloumba catchment in all years (pre- and post-disturbance). Turbidity was also markedly affected in 1987 when gold-bearing ore was removed from a site on the Bundaroo catchment, with a significant difference occurring between turbidity measures above and below the site and significantly larger juveniles occurring in Bundaroo Creek in that season. Growth data did not correlate well with turbidity data, probably because of the inability of the equipment available at the time to detect high run-off episodes associated with mining at least. The technology is now able to record such events. It is unlikely selective logging activities produced higher than normal stream turbidity levels during high run-off episodes; this is partly because most of the unlogged vegetation buffers along permanent and semi-permanent watercourses within the North Booloumba catch- ment were considerably wider than the 10 m wide buffers that were standard logging practice at the time. Rainfall data did not correlate well with turbidity data as turbidity is greatly affected by episodic rainfall events within each catchment and may go undetected at the Kenilworth Forestry Station. Similarly rainfall could not be correlated with growth or abundance data, which serves further only to highlight the difficulties we experience in attempting to mea- sure environmental and life history characteristics on the same time/space scales. While this project provides background information about the habitat requirements, fluctuations in growth rates and abundance of E. hystricosus within the Conondale Unit of the Conondale Ranges, the patchiness of data collected over many years makes it diffi- cult to make definitive causal connections between life history parameters (growth, abun- dance) and changes in the environment due to anthropogenic effects (forestry, mining). In terms of assessing whether E. hystricosus has been useful as a monitoring tool for detecting adverse environmental change, we can only say that it is unclear even still. It is apparent however, that E. hystricosus has the same characteristics (i.e. it is easy to mark individuals; it shows sedentary behaviour; it has limited physiological tolerances; and it has a long life span (Borsboom and Kehl, unpublished data)) that make E. spinifer an ideal organism for bio-monitoring (Merrick 1997). However, suffice it to say that any monitoring program requires long-term commitment in all its respects and it is critically important that the scale at which measurements are taken should be considered carefully so that no fine scale relationships are overlooked in the final synthesis. We recommend further studies and monitoring of E. hystricosus at other sites with- in the range of this species, to further assess its ecological requirements and any poten- tially adverse impacts from anthropogenic influences (Merrick 1995). Prior to formulat- ing specific conservation strategies for E. hystricosus the recent management findings on other large Euastacus species of recreational importance should be considered (Barker 1990; Honan and Mitchell 1995; Merrick 1997). Populations of E. hystricosus are rela- tively well conserved within state forests and national parks in Queensland. Illegal har- vesting is a possible threat, however, within the Conondale National Park a number of access roads to water courses with E. hystricosus have recently been closed to the public. Monitoring of the species and control of illegal harvesting will do much to ensure the cray’s security on state forest. Proc. LINN. Soc. N.S.W., 119. 1998 86 JUVENILE EUASTACUS HYSTRICOSUS ACKNOWLEDGEMENTS Project initiation and fieldwork were carried out primarily by Adrian Borsboom, Chris Corben, Greg Cooper and John Kehl, with analyses undertaken by Rina Lloyd, Nadya Lees, and the senior author using Systat, Microsoft Excel and Statistica computer packages. Janet Chasely (Griffith University) and Anita Smyth (University of Queensland) advised on appropriate statistical analyses. Thanks to Murray Willson and Kristy Willett for help with preparation of figures, Benjamin Hamley for preparation of the manuscript and to staff of the Kenilworth Forestry Office for their assistance. The study has been funded by allocations from the Forest Protection and Management and Integrated Catchment Management sub-programs of the Queensland Government (1982-97). Thanks to J. Merrick and P. Turvey for their constructive editorial comments. REFERENCES Anon. (1995). ‘Environmental Monitoring and Performance. Best Practice Environmental Management in Mining’. (Environment Protection Agency, Commonwealth of Australia). Barker, J. (1990). Spiny freshwater crayfish management strategy. Fisheries Management Report (34), 1-17. Department of Conservation and Environment, Melbourne. Bernstein, B.B. and Zalinski, J. (1983). An optimum sampling design and power test for environmental biolo- gists. Journal of Experimental Marine Biology and Ecology 16, 35-43. Borsboom, A. (1998). Aspects of the biology and ecology of the Australian freshwater crayfish, Euastacus urospinosus (Decapoda: Parastacidae). This volume. Bunn, S.E. (1995). Biological monitoring of water quality in Australia: workshop summary and future direc- tions. Australian Journal of Ecology 20 (1), 220-227. Faith, D.P. and Norris, R.H. (1989). Correlation of environmental variables with patterns of distribution and abundance of common and rare freshwater macro-invertebrates. Biological Conservation 50, 77-98. Honan, J.A. and Mitchell, B.D. (1995). Catch characteristics of the large freshwater crayfish Euastacus bispinosus Clark (Decapoda: Parastacidae), and implications for management. Freshwater Crayfish 10, 57-69. Merrick, J.R. (1993). ‘Freshwater crayfishes of New South Wales’. (Linnean Society of New South Wales, Sydney). Merrick, J.R. (1995). Diversity, distribution and conservation of freshwater crayfishes in the eastern highlands of New South Wales. Proceedings of the Linnean Society of New South Wales. 115, 247-258. Merrick, J.R. (1997). Conservation and field management of the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), in the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales. 118, 217-225. Morgan, G.J. (1988). Freshwater crayfish of the genus Euastacus Clark (Decapoda: Parastacidae) from Queensland. Memoirs of the Museum of Victoria 49 (1), 1-49. Morgan, G.J. (1991). The spiny freshwater crayfish of Queensland. Queensland Naturalist. 31 (1-2), 29-36. Norris, R.H. and Norris, K.R. (1995). The need for biological assessment of water quality: Australian perspec- tive. Australian Journal of Ecology 20 (1), 1-6. Riek, E.F. (1951). The freshwater crayfish (Family Parastacidae) of Queensland, with an appendix describing other Australian species. Records of the Australian Museum 22, 368-388. Sokal, R.R. and Rolf, F.J. (1981). ‘Biometry’. (Freeman, San Francisco). Stewart-Oaten, A., Murdoch, W.W. and Parker, K.R. (1986). Environmental Impact Assessment: “‘Pseudo-repli- cation” in time? Ecology 67, 929-940. Turvey, P. and Merrick, J.R. (1997a). Reproductive biology of the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales. 118, 131-155. Turvey, P. and Merrick, J.R. (1997b). Population structure of the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales. 118, 157-174. Turvey, P. and Merrick, J.R. (1997c). Diet and feeding in the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales. 118, 175-185. Turvey, P. and Merrick, J.R. (1997d). Moult increments and frequency in the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales. 118, 187-204. Turvey, P. and Merrick, J.R. (1997e). Growth with age in the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales. 118, 205-215. Proc. LINN. SOC. N.S.W., 119. 1998 Aspects of the Biology and Ecology of the Australian Freshwater Crayfish, Euastacus urospinosus (Decapoda: Parastacidae) ADRIAN BORSBOOM (Communicated by J.R. Merrick) Resource Sciences Centre, Queensland Department of Natural Resources, PO Box 631, Indooroopilly, Qld 4068 Borssoom, A. (1998). Aspects of the biology and ecology of the Australian freshwater cray- fish, Euastacus urospinosus (Decapoda: Parastacidae) Proceedings of the Linnean Society of New South Wales 119, 87-100. The range of the upland freshwater crayfish, Euastacus urospinosus, has been extend- ed to the Conondale Ranges (southeastern Queensland) where it occurs in creeks as well as bank burrows in rainforest at 450-550 m altitude. During sampling and trapping from 1982-1994 a total of 685 individuals were examined and aspects of reproduction, population structure, growth and habitat usage investigated. Breeding females range from 33.8—51.8 mm ocular carapace length (OCL), appear to breed annually, carry a mean of 51 (3-119) eggs and average 31 (3-93) young. Mating appar- ently commences in April with males moving considerable distances (>20 m) to burrows housing mature females to pair with them. Eggs are laid about May or June, incubated for four to five months, hatch in late October or November and young released in December in the creeks. Adults only live in bank burrows and immature individuals occur predominantly in the creek. The mean OCL for crayfish in the creeks is 11.2 mm (n = 492) and 30.3 mm (n = 162) in bank burrows. The smallest free-living individual had an OCL of 5.5 mm and the largest, a male, measured 54.1 mm OCL and weighed 84.5 grams. It is estimated that females take approximately six years to reach breeding size. A simple trapping method using black plastic tubing inserted at entrances, enabled regular capture of crays in burrows. The resulting mark-recapture program has shown that burrows are normally only occupied by one individual, except during mating in April; howev- er, trapping success is correlated with water temperature, with least success in winter. OCL has also been demonstrated to increase with burrow entrance diameter. Manuscript received 14 October 1997, accepted for publication 15 January 1998. KEYWORDS: breeding seasonality, burrows, Euastacus urospinosus, growth, maturation, population structure, reproduction, trapping . INTRODUCTION There has been no detailed account of any aspect of the biology and ecology of the Australian crayfish Euastacus urospinosus since its description by Riek (1956), and, prior to these studies, no mature females had been collected (Morgan 1988, 1991). Morgan (1988) provided a comprehensive morphological description of the species, and the maxi- mum ocular carapace length (OCL) of specimens examined was 36.7 mm. Morgan (1988) reported that E. urospinosus was restricted to tributaries of Obi Obi Creek in the Mary River Drainage Basin between Maleny and Mapleton in south-eastern Queensland, where it was found in sub-tropical rainforest above an altitude of 240 m; however, Morgan (1989) believed that the distribution of E. urospinosus might also extend to the Conondale Ranges. The studies reported here originated as part of a broader research program on the impacts of forestry on water quality and aquatic faunas. The area under investigation was state forest until incorporated into the Conondale National Park in 1995. Gold prospect- ing and mining had occurred along the creeks and selective logging has taken place in Proc. LINN. SOc. N.S.W., 119. 1998 88 BIOLOGY AND ECOLOGY OF EUASTACUS UROSPINOSUS Scale 0 ee | Kilometre Kenilworth e Forest Stations Figure 1. E. urospinosus study area. Key to symbols: 4 mark recapture site, @creek section sampled, and __ rainforest. Proc. LINN. SOc. N.S.W., 119. 1998 A. BORSBOOM 89 the Booloumba and North Booloumba Creek catchments. The Bundaroo Creek catch- ment has no history of logging, although cedar-cutters may have taken some timber earli- er this century. Aside from verifying the suspected extended range of EF. urospinsosus and docu- menting major habitat parameters, this paper has the following objectives: to investigate reproductive maturation, fecundity and seasonality; to report on population structure as well as growth; and to describe habitat usage. MATERIALS AND METHODS Study Area The study area lies in the Conondale Ranges NNW of Brisbane at approximately 26°42'S, 152°36’E. Crayfish were studied on Booloumba Creek and its tributaries North Booloumba Creek and Bundaroo Creek at 450-550 m altitude. Booloumba Creek drains to the Mary River. Mark-recapture studies were conducted at two sites, one each on Booloumba Creek and Bundaroo Creek (Fig. 1). Data for instream crayfish were collect- ed at various sites on Booloumba Creek or its tributaries, and include opportunistic records from sites where Euastacus hystricosus was under study (Smith et al. 1998). E. hystricosus is sympatric with E. urospinosus in this area. Booloumba Creek and its tributaries can be broadly described as permanent, mon- tane, rocky watercourses with slow basal flows. During periods of basal flow the creeks consist of a series of pools interconnected by glides, runs or riffles. Usually pools are less than 1.2 m deep and 30 m in length, occasionally there are larger and deeper pools (<150 m long: <3.0 m depth). Some sections are dominated by larger rocks and boul- ders, with occasional sections where the flow is over exposed bedrock with cascades or waterfalls when the gradient is steeper. Pool width varies but is generally less than 10 m and rarely over 20 m. Substrates in the larger pools are usually a mosaic of slopes, flats and bars of gravel, stones and rocks, large loose rocks, boulders, logs and accumulations of decaying organic matter. These lie on either bedrock or a bed of gravel, stones and rocks. Riffles, runs and glides generally have substrates of loose rocks and stones lying on a bed of gravel, rocks and stones. Vegetation within the study area is mainly rainforest and wet sclerophyll forest, with or without eucalypt emergents. The upper tree storey includes Archontophoenix cunning- hamiana (Piccabeen Palm), Argyrodendron actinophyllum (Tulip Oak), A. trifoliatum (Booyong), Auracaria bidwillii (Bunya Pine), Caldcluria paniculosa (Rose-leaf Marara), Castanospermum australe (Black Bean), Diopyrus pentamera (Black Myrtle), Eleocarpus grandis (Blue Quandong), Ficus watkinsiana (Strangler Fig), Gmelina leichardtii (White Beech), Lophostemon confertus (Brush Box), Planchonella australis (Black Plum), Sloanea woollsii (Yellow Carabeen) and Syzigium spp. Emergents were mainly Eucalyptus grandis (Flooded Gum) in gullies, while on upper slopes and ridges E. gum- mifera (Red Stringybark) and E. microcorys (Tallowood) were the main emergents. Rainforest dominates along the creeks and in places the palm, Archontophoenix cunninghamiana, forms stands with little ground vegetation. Vegetation on the creek verges is sparse where there are large expanses of exposed bedrock. Ground cover can be dominated by tufted clumps of mat rush, Lomandra longifolia, wherever boulders domi- nate a broader floodplain and channel. The exotic weeds Lantana camara (Lantana) and Eupatorium riparium (Mist Flower) are also present. Temperatures Temperatures were monitored at both mark-recapture sites using maximum-mini- mum thermometers. In Bundaroo Creek readings were taken in shallow, moving water in Proc. LINN. Soc. N.S.W., 119. 1998 90 BIOLOGY AND ECOLOGY OF EUASTACUS UROSPINOSUS a well shaded riffle; in Booloumba Creek measurements were taken in shallow, well shad- ed moving water at the confluence of a riffle with a pool/glide. Ground water tempera- tures were estimated by burying screw-top 4 litre plastic containers of water each with a thermometer inside, with the top of the container about 0.2 m underground. Readings were taken by digging down to the top of the container, unscrewing the lid, and then, with minimal handling, reading and resetting the thermometer while keeping it under water. The lid was re-screwed and the top of the container again covered with soil and leaf litter. Measurement, Trapping, Marking and Sampling Size of captured individuals was determined by the ocular carapace length (OCL), measured from the posterior orbit margin to the dorsal posterior carapace margin, using dial vernier callipers (nearest 0.1 mm). Weights in the field were taken (nearest 0.25 g) using a Pesola spring balance. In preliminary field surveys it was observed that FE. urospinosus would often sit just inside the burrow entrance at night or late in the day. To take advantage of this behaviour a section of flexible black plastic tubing (~ 0.40—0.45 m long), of a diameter similar to the entrance, was pushed 30-50 mm into the burrow and bent flat to the ground using a rock on site. This tubing was left permanently in place during the mark- recapture study and was quickly accepted by crays as part of the burrow system. Individuals continued to sit just inside their ‘extended’ burrow and were captured by pulling the tubing from the burrow at night. After examination, measurement and tagging if required, crayfish were released into their burrow and the tubing re-inserted. Individuals caught in the mark-recapture study were tagged using notching pliers by cut- ting one or more small v-shaped notches in either the telson and/or uropods. Although the notches grew over, marks were always clearly discernible after one moult and still discernible enough after two moults to identify the individual. Moult occurrence was recorded and where possible marked crays were re-marked after each moulting. E. urospinosus caught in creeks were not marked, except for a few caught inciden- tally in a E. hystricosus study, but regular sampling was carried out on circular 1 m plots in riffle areas of Boolumba, North Booloumba and Bundaroo Creeks. Plots were sampled by lifting and removing stones or small rocks and crays captured by hand or with a small dip net. All captives were measured, weighed and sexed where possible. After sampling, the rocks and stones were placed back into the plot and the residents released throughout the plot. Crays were also caught in the creeks by searching under stones and rocks on three bank/creek cross-section survey plots, one on Booloumba Creek, two on Bundaroo Creek. All burrows in the three cross-sectional survey plots were excavated as well as a small number of burrows from the mark-recapture study. RESULTS This extended sampling program has confirmed the presence of substantial resident populations of E. urospinosus in the Conondale Ranges. During the period December 1982 to July 1994 some 685 individuals were captured and examined in the study area (Fig. 1). A total of 956 records were generated as some crays were caught more than once in the mark-recapture study. Reproduction All females (n = 143) below 33.2 mm OCL had flat, non-inflated gonopores; where- as 48 females 233.2 mm OCL had inflated gonopores and 21 carried eggs or juveniles. The smallest female with eggs or young was 33.8 mm OCL and 18.0 grams, the largest Proc. LINN. SOC. N.S.W., 119. 1998 A. BORSBOOM 9] JUL 26th o oO Te) > 2) a <= iw ra a) LL << 25 20 xr ” ~ 15 O LL O ~ Lu = 10 > z | HH | oO oo oO wt oOo wo OH Sion Sie ae Aa AT enn © ee) Ge Lg N Syn eSa ew Z i 8 3 = ” Zz JUN 15-16 | | (ep) Ni; mR oe O Lid Q MAR 11-20 TIME PERIOD Figure 2. Reproductive status of mature E. urospinosus females (+33.8 mm OCL or with swollen gono- ye: Key to symbols: H individuals carrying larvae or juveniles; MM individuals car rying eggs (in berry): individuals not in berry. Proc. LINN. SOC. N.S.W., 119. 1998 92 BIOLOGY AND ECOLOGY OF EUASTACUS UROSPINOSUS y = 3.9725x - 112.75 R? = 0.4611 NUMBER OF EGGS 35.0 40.0 45.0 50.0 55.0 : OCL (mm) Figure 3. The relationship between OCL and numbers of eggs carried by E. urospinosus females. recorded female was 51.8 mm OCL (63 g). It appears inflation of the gonopores correlates strongly with maturation. Mature females were only found in bank burrows and not in the creeks. Figure 2 graphically presents the yearly reproductive status of mature females. The data indicate that E. urospinosus is a winter brooder with a fixed annual cycle and a single synchronised release of juveniles in December, but the trigger for release is unknown. The earliest captures of females carrying eggs were late May (n = 3) and mid- June (n = 4). Egg incubation is 4-5 months. By the later half of November 86% of breed- ing females (n = 7) were carrying juveniles. No mature females were carrying eggs from 17 December to 10 April (n = 91), or eggs/juveniles from 8 January to 10 April (n = 67). All females caught over either two consecutive breeding seasons (n = 5) or three consecu- tive breeding seasons (n = 2), were carrying eggs or juveniles in each season; but not all mature females bred successfully each year. About one third of mature females recorded from late July to late November were not carrying eggs or juveniles. The mean number of eggs carried by females when first captured during a breeding season was 51 (n = 25; range 3-119). The linear regression between OCL and the num- ber of eggs carried is illustrated in Fig. 3; the larger the female the greater the number of eggs. The mean number of juveniles carried in November is 31 (n = 7; range 3—93). In the mark-recapture study one female was monitored occupying the same burrow system for a 22 month period (Sept. 1984 to May 1986) and for the three breeding peri- ods covered was carrying 71, 70 and 73 eggs respectively. In the second breeding season the female was not carrying eggs on 11 March, but when recaptured on 9 October had moulted and was in berry. In the interim period a mature male (37.6 mm OCL) had been captured (30 April) in the female’s burrow. It had moved from a burrow 23 m away in which it had been captured on 10 April; this male was never recaptured or seen again in the female’s burrow. The following year the female was not carrying eggs on 19 March, but on 20 May it had moulted and was carrying eggs again. During the intervening peri- od a different mature male (47.7 mm OCL) had been captured in the burrow (22 April). On 20 May the female’s burrow was dug out and only the female was found in the bur- Proc. LINN. Soc. N.S.W., 119. 1998 A. BORSBOOM 93 row system. Prior to capture in the female’s burrow in April this male had been caught on 20 March in a burrow 5.75 m away. These observations suggest that a mature male will pair and mate with a mature female in her burrow system around April of each year. Population Structure and Growth The smallest free-living E. urospinosus had an OCL of 5.5 mm, the largest female recorded was 51.8 mm OCL (63.5 g) and the largest male was 54.1 mm OCL and 84.5 grams. At sites shown (Fig. 1) crays were caught in the creeks at <0.5 m depth under stones, rocks and logs in riffles, runs, glides and the shallow reaches of pools; larger indi- viduals often had a shallow burrow system under rocks. Crays in the creek were generally small (11.2 mm mean OCL: n = 492, SD = 4.87), ranging from 5.5 mm OCL to a female of 29.7 mm OCL (14.2 g); the largest male was 26.3 mm OCL (8.4 g). E. urospinosus also lived in burrows in the creek banks. Burrows were found up to 6 m from the basal flow water mark, especially where banks had a broader lower bench with or without flood bars of gravel, stones and rocks. There was often a concentration of burrows where the lower bank bench ended in a sharp rise. Mean OCL of bank burrow populations was 30.3 mm OCL (range 11.9-54.1 mm; n = 162; SD = 9.36). The ratio of males to females for crays captured in burrows was 1:1.16 (n = 162), and was not signifi- cantly different from 1:1 (x? = 0.88; d.f. = 1; p >0.1). The mean OCL of crays in the creek was highly significantly different (p <0.001) to the mean OCL for crays living in burrows in the banks adjacent to the creek. The Mann-Whitney U test was used for this analysis as the variances were not homogeneous. Size distribution of pooled OCL data for E. urospinosus caught in three creeks dur- ing this study were plotted as bi-monthly frequency histograms (Fig. 4). The size classes in the December—January period of newly released juveniles were clearly distinct from one year old crays (Fig. 4a), but the separation between one and two year olds in that peri- od is not distinct, and even less so for three year olds, as the sample of individuals over 16 mm OCL is low. Subsequent bi-monthly plots (Fig.4 b—f), indicate that individuals between 11.0 and 16.8 mm OCL (in December—January) were probably one year old. This suggested that E. urospinosus had a mean OCL of 13.7 (n = 18) at 12 months of age; at two years individuals were ~ 18-19 mm OCL, and three year old individuals were ~ 22-24 mm OCL. From these size/abundance values and moult frequency data, from mark-recapture individuals, it was possible to infer an age at which females reached maturity. Individuals with an OCL 24.1—30.8 mm (n = 7) moulted twice per year, but there was a transition from twice to once yearly moulting for crays from 30.8—33.6 mm OCL (n = 8). All larger indi- viduals (233.7 mm OCL), where yearly moult data were available (n = 27), moulted once per year. To determine an age for female maturity, moult increments for crays 222.4 mm OCL were plotted as a percentage of pre-moult OCL (Fig. 5) and the relationship which most accurately reflected the data was the exponential y = 54.732 e°°”* (R® = 0.684). According to this relationship an individual of 23 mm OCL would moult a further five times to reach a size of 33.2 mm OCL (size at which at which female gonopores become inflated). As crays 24.1-30.8 mm OCL moult twice a year and larger animals (30.8—33.8 OCL) moult annually, it is reasonable to suppose a female of 23.0 mm OCL and three years old, would take on average about another three years to reach maturity. Bank Burrows A total of 70 E. urospinosus (11.9-47.0 mm OCL) were dug out of bank burrows. The sample included an unsexed specimen as well as 37 females and 32 males. No more than one individual was retrieved from any burrow system. Forty-eight crays were dug out in February, 5 in April, 6 in May, 5 in June and 6 in September. All excavated burrows Proc. LINN. SOc. N.S.W., 119. 1998 94 BIOLOGY AND ECOLOGY OF EUASTACUS UROSPINOSUS NUMBER INDIVIDUALS CAPTURED RMDAAAAAARARAARHRAAAARAARARAARAAR RA DR @ oor DO DMWDOerTrH iN Oe TFT HOR DO DO TiN HMHtFT HN Oo KF GB HD FS GG) Gy (6) Gy Am Sa Np NE Se Sn SS Ga Gun GaN Gully Gal Gao Gal Gal Ss See DO SS @ SQ 1G ae S&S So Se eG ee SE 42°92 e759 7°22 ¢ or Nn mOoO tT NH OR DOD DW Oo Kr NCUcUMDhmUCUMSTGDLC >) i) i=) o So o o 1 1 1 1 1 1 1 1 1 1 1 ' D 1 ! 1 ' ' D t i BS OS 88 Go eo ol QS oe oS Sooo Se eS SoSua GS a2 Ss So = N o wt wo o Ld (<<) for) o = N o + w wo is on oo So oo So > > >) > OCL SIZE CLASSES (mm) Figure 4 continued. Pooled OCL size classes for E. urospinosus caught in Bundaroo, Booloumba and Nth Booloumba Creeks over 2 month intervals: (f) October and November. reached the water table and had up to three entrances; they were <1.3 m deep and $1.5 m across. Adults typically occupied burrows with one to two chambers at the bottom, one of which was always filled or partially filled with ground water. Chambers were normally 0.25—0.30 m long and 75-125 mm in diameter, but could be as long as 0.95 m. There was usually at least one blind tunnel and sometimes a closed chimney in a burrow system. Whenever an individual was caught undisturbed in a burrow system it was often sitting at the edge of, or partly in, the water in a chamber. Linear regression showed a positive rela- tionship between OCL and burrow entrance diameter. For crays 14.7-54.1 mm OCL (n=109) the burrow diameter increased with OCL (y = 1.0096x — 0.9416; R* = 0.741). Trapping and Temperature Figure 6 summarises tube-trapping capture data and creek and groundwater tem- peratures at two mark-recapture sites. Creek water was below 20°C for most of the year and reached a maximum of 22.0-22.5°C. Minimum groundwater temperature (at burrow depth) in mid-winter was up to 4.5°C warmer than minimum water temperature in the creek riffle zones. Clearly E. urospinosus living in burrows showed little activity at their burrow entrances when creek and groundwater temperatures were at their lowest. DISCUSSION E. urospinosus belongs to a group of eight smaller (<55 mm OCL), poorly known Queensland Euastacus species, three of which in south-east Queensland are considered similar in morphology and size (Morgan 1991). E. urospinosus in the Conondale study area occurs mainly in rainforest where it lives in cooler waters at a higher altitude, similar to most other Queensland Euastacus species (Morgan 1991; Riek 1969). The maximum sizes recorded (female 51.8 mm OCL, 63.5 g : male 54.1 mm OCL, 84.5 g) during these studies make E. urospinosus the largest of the eight smaller, weakly spined Queensland Euastacus. Proc. LINN. SOc. N.S.W., 119. 1998 A. BORSBOOM 97 16.0 14.0 12.0 10.0 “ . : y = 54.732e0°"™ a Sa R? = 0.684 = > ° °o MOULT INCREMENT (% OF PREMOULT OCL) Ny © ° r=) 25.0 30.0 35.0 40.0 45.0 50.0 55.0 PRE-MOULT OCL (mm) Figure 5. Relationship of moult increment (%) to pre-moult OCL in E. urospinosus 222.4 mm OCL. Parastacid crayfish normally have either a relatively short egg incubation period during the warmer months (summer brooders), or a long incubation period over winter (winter brooders) (Honan and Mitchell 1995). This study indicates E. urospinosus is a winter brooder, similar to the sympatric Euvastacus hystricosus (Borsboom, unpublished data), and a number of other Euastacus species listed in a reproductive review table in Turvey and Merrick (1997a). E. urospinosus has a lower fecundity (119) than the compa- rably-sized E. australasiensis (155), E. keirensis (184) and E. yanga (164) (Turvey and Merrick 1997a). The reason for E. urospinosus having a lower fecundity is unclear. It may relate to mortality levels, as breeding individuals live in burrows where they are not subject to eel predation. In addition, immature E. urospinosus are probably less at risk from eel predation as they rarely move about freely at night in the creek, unlike the sym- patric immature EF. hystricosus. Breeding in E. urospinosus follows a fixed annual cycle with a single synchronised release of juveniles in December. It is suggested that some unknown environmental vari- able(s) triggers a female in December to leave its burrow, move to the creek, release young then return to its burrow. In support of this hypothesis, no free-living juveniles have been found in the burrows of mature females that have been excavated, or captured sharing a mature female’s burrow. Additional supportive anecdotal observations follow. In mid-December 1983 a female carrying several release-size juveniles was caught at night in rain moving freely along Bundaroo Creek at the edge of the basal flow. This individual was approximately 1.2 m from any burrow entrance, which is the furthest any adult was observed away from a burrow entrance. The only other records of crays away from a burrow entrance were two separate instances of individuals feeding in leaf litter approximately 0.5 m and 0.6 m from burrow entrances. Although triggers for mating, egg laying and release of juveniles are unknown in E. urospinosus, water temperature and photoperiod cycles have been suggested for other species (Aiken 1969; Turvey and Merrick 1997a); however, Honan and Mitchell (1995) believe control of the reproductive cycle may involve more than one periodic environ- Proc. LINN. SOC. N.S.W., 119. 1998 98 BIOLOGY AND ECOLOGY OF EUASTACUS UROSPINOSUS TEMP. °C/CRAY TRAPPING CAPTURE INDEX 6/08/84 25/09/84 14/11/84 3/01/85 22/02/85 13/04/85 2/06/85 22/07/85 10/09/85 30/10/85 19/12/85 = — ied nN o a Oo a TEMP. °C/ CRAY TRAPPING CAPTURE INDEX on 6/08/84 , 25/09/84 14/11/84 3/01/85 22/02/85 13/04/85 2/06/85 10/09/85 30/10/85 19/12/85 Figure 6. Minimum groundwater temperature and riffle zone minimum and maximum water temperatures for E. urospinosus trapping sites compared with the crayfish tube-trapping capture index: (a) Booloumba Creek; (b) Bundaroo Creek. Key to symbols: ® = groundwater temperature (min); B = riffle water temperature (min); ) = riffle water temperature (max); J = trapping capture index. Proc. LINN. SOC. N.S.W., 119. 1998 A. BORSBOOM 99 mental variable. Rain, high humidity and/or a rise in stream levels may be important components of the triggering mechanism for the synchronised release of juvenile E. urospinosus in December. It has been estimated that female E. urospinosus take approximately 6 years to reach reproductive maturity, but there is no information on how long other comparably- sized Euastacus take to reach maturity. E. armatus, E. bispinosus and E. spinifer which grow considerably larger, take 6—9, 8-11 and 5-6 years respectively (Turvey and Merrick 1997d). So for a crayfish of this size (up to 84.5 g), six years is a long matura- tion period; similar sized Cherax destructor, can commence reproducing in less than a year (Merrick 1993). The type of partitioning suggested in E. urospinosus populations, where adults live in bank burrows and immature individuals in the creeks, is supported by the absence of adult captures in shallow and deep pools where E. hystricosus trapping was undertaken, and by the lack of observations of E. urospinosus adults in the creeks to depths <2.5 m, although field inspections occurred at all times of the day over many years. This form of population segregation (adults in creek bank burrows and juveniles in creeks) has not been reported to date in any other Euastacus species. In the larger species E. armatus, E. bispinosus and E. spinifer, adults live and feed in watercourses (Hogger 1988; Honan and Mitchell 1995; Morgan 1986; Turvey and Merrick 1997b, c). E. jagara, E. monteithorum and E. setosus, which are similar in morphology and size to E. urospinosus (Morgan 1991), also construct bank burrows (Borsboom, unpublished data). Whether they too have adults living permanently in bank burrows requires further investigation. E. urospinosus burrows can be classed as Type 2 sensu Horwitz and Richardson (1988), but the capture of individuals in this burrow type by tube-trapping could have wider application. This technique allows the regular recapture of burrowing crayfish with minimal stress and no disturbance of burrows or habitat, and has potential for many other cray species. ACKNOWLEDGEMENTS Field studies were designed and implemented by the author as part of a broader research program head- ed by J. Kehl on the impact of logging on stream water quality and stream faunas. Special thanks to J. Kehl for providing constructive comment on experimental design for the mark-recapture study. Thanks also to J. Merrick and P. Turvey for providing constructive comments on the submitted manuscript. REFERENCES Aiken, D.E. (1969). Ovarian maturation and egg laying in the crayfish Orconectes virilis: influence of tempera- ture and photoperiod. Canadian Journal of Zoology 47, 931-935. Hogger, J.B. (1988). Ecology, population biology and behaviour. In ‘Freshwater crayfish: biology, management and exploitation’. (Eds D.M. Holdich and R.S. Lowery) pp. 114-129. (Croom Helm Ltd., London). Honan, J.A. and Mitchell, B.D. (1995). Reproduction of Euastacus bispinosus Clark (Decapoda: Parastacidae), and trends in the reproductive biology of freshwater crayfish. Marine and Freshwater Research 46 (2), 485-499. Horwitz, P.H.J. and Richardson, A.M.M. (1988). An ecological classification of the burrows of Australian cray- fish. Australian Journal of Marine and Freshwater Research 37, 237-242. Merrick, J.R. (1993). ‘Freshwater crayfishes of New South Wales’. (Linnean Society of New South Wales, Sydney). Morgan, G.J. (1986). Freshwater crayfish of the genus Euastacus (Decapoda, Parastacidae) from Victoria. Memoirs of the Museum of Victoria 47, 1-57. Morgan, G.J. (1988). Freshwater crayfish of the genus Euastacus Clark (Decapoda, Parastacidae) from Queensland. Memoirs of the Museum of Victoria 49 (1), 149. Morgan, G.J. (1989). Two new species of the freshwater crayfish Euastacus Clark (Decapoda: Parastacidae) from isolated high country of Queensland. Memoirs of the Queensland Museum 27 (2), 555-562. Proc. LINN. Soc. N.S.W., 119. 1998 100 BIOLOGY AND ECOLOGY OF EUASTACUS UROSPINOSUS Morgan, G.J. (1991). The spiny freshwater crayfish of Queensland. Queensland Naturalist 31 (1-2), 29-36. Riek, E.F. (1956). Additions to the Australian freshwater crayfish. Records of the Australian Museum 24, 1-6. Riek, E.F. (1969). The Australian freshwater crayfish (Crustacea: Decapoda: Parastacidae), with descriptions of new species. Australian Journal of Zoology 17, 855-918. Smith, G.C., Borsboom, A., Lloyd, R., Lees, N. and Kehl, J.C. (1998). An investigation of the habitat, growth and abundance of juvenile Giant Spiny Crayfish, Euastacus hystricosus (Decapoda: Parastacidae), in the Conondale Ranges, south-east Queensland. This volume. Turvey, P. and Merrick, J.R. (1997a). Reproductive biology of the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales 118, 131-155. Turvey, P. and Merrick, J.R. (1997b). Population structure of the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales 118, 157-174. Turvey, P. and Merrick, J.R. (1997c). Diet and feeding in the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales 118, 175-185. Turvey, P. and Merrick, J.R. (1997d). Growth with age in the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales 118, 205-215. Proc. LINN. SOC. N.S.W., 119. 1998 Limited Usage of Freshwater Crayfishes (genus Euastacus) by Aborigines in Eastern New South Wales: Records and Comments J.L. KOHEN'! AND J.R. MERRICK? 1School of Biological Sciences, Macquarie University, North Ryde NSW 2109 and 2Graduate School of the Environment, Macquarie University, North Ryde NSW 2109. KOHEN, J.L. AND MERRICK, J.R. (1998). Limited usage of freshwater crayfishes (genus Euastacus) by Aborigines in eastern New South Wales: records and comments. Proceedings of the Linnean Society of New South Wales 119, 101-105. Australian Aborigines utilised freshwater crayfishes as food in many areas and these macro-invertebrates had substantial cultural significance. To investigate the extent of this fishery in eastern New South Wales, where Aboriginal populations are thought to have con- centrated, literature and database surveys of contents of selected Holocene Aboriginal sites have been undertaken; but few remains of Euastacus species, known to be abundant in this region, are recorded. In the light of recent scientific studies of Euastacus, the possible local impacts of har- vesting on these crays are outlined and the paucity of remains at Aboriginal sites is consid- ered to be due to: inadequate sampling or sorting of existing collections; poor preservation conditions; dispersal or destruction by scavengers; very low utilisation and correspondingly few recognisable remnants. Manuscript received 23 July 1997, accepted for publication 5 January 1998. KEYWORDS: Aboriginal utilisation, eastern Australian highlands, Euastacus, food resources, macro-invertebrates. INTRODUCTION Australian Aborigines are known to have utilised large numbers of animal species as food (Isaacs 1987). Crayfishes are one of the largest groups of edible macro-inverte- brates in inland areas and the Australian crayfish fauna is very diverse (Merrick 1993). Crayfishes were harvested in many areas and described as a favoured food as well as a staple of the diet in some regions (Horwitz and Knott 1995; Isaacs 1987); however, there is relatively little information about harvesting techniques. General comments about col- lection of crays include observations of digging from burrows around ponds or swamps (Campbell 1978), as well as catching by hand from under large stones in rivers when ley- els were low (Flood 1980); but more specialised trapping methods (woven baskets with baits, sections of hollow logs) were also used (Horwitz and Knott 1995). Crayfishes (or yabbies) also had cultural importance. For example, there were yabby totems, places named after yabbies and crayfish ‘ancestors’; a yabby clan is also reported to have lived in the Mt. Gambier—Grampian Range area (Horwitz and Knott 1995). Although Aboriginals utilised crayfishes as a food source over wide areas of New South Wales (Horwitz and Knott 1995; Versteegen and Lawler 1997), no systematic archaeological studies have been undertaken to assess the extent of this fishery. It is also clear that a large majority of the crayfish species in N.S.W. are restricted to the highlands or eastern coastal plain (Fig. 1) and some species have small total ranges (Merrick 1993; Morgan 1997). The only reports relating specifically to Euastacus in eastern New South Wales are: E. armatus being caught in the evenings in the Murrumbidgee, Yass and Proc. LINN. Soc. N.S.W., 119. 1998 102 LIMITED ABORIGINAL USAGE OF EUASTACUS Za aa McPherson Ra — Zin, Se LD : 7— * Richmond \7~ Gs Mt Warning [ ( \ / ees Tenterfield Bellingen R. Macleay R. Newcastle | , 0 f Katoombae | | Belmore Falls Fitzroy Falls Ye cj — Canberra /94 ae / Key to species: (1) E. armatus; (2) E. australasiensis; (3) E. bidawalus; (4) E. brachythorax; (5) E. clarkae; (6) E. claytoni; (7) E. crassus; (8) E. dangadi; (9) E. dharawalus; (10) E. gamilaroi; (11) E. gumar; (12) E. guwinus; (13) E. hirsutus; (14) E. neohirsutus; (15) E. polysetosus; (16) E. reductus; (17) E. rieki; (18) E. simplex; (19) E. spinichelatus; (20) E. spinifer; (21) E. sulcatus; (22) E. suttoni; (23) E. valentulus; (24) E. yanga. Figure 1. Natural ranges of 24 Euastacus species in eastern New South Wales over the region for which collection records have been examined. A further six crayfish species of two other genera are also present in this region. Based on Merrick (1995) and Morgan (1997). Proc. LINN. Soc. N.S.W., 119. 1998 J.L. KOHEN AND J.R. MERRICK 103 Tumut Rivers; small crays (almost certainly Ewastacus) being found in the highest per- manent streams of the southern highlands around Mount Kosciusko (Flood 1980). Another species, Euastacus sulcatus, which occurs in north-eastern New South Wales has been recorded at an Aboriginal site in south-eastern Queensland (Hall 1986). In view of this faunal concentration adjacent to areas where Aboriginal populations are claimed to have undergone substantial intensification for several thousand years (Lourandos and Ross 1994), these studies were initiated to ascertain the level of utilisa- tion of selected Euastacus species by means of faunal analysis. To date faunal analyses, in this area, have focused on vertebrate remains — especially the larger, more robust components (Godfree 1995; Owen and Merrick 1994a, b). MATERIALS AND METHODS The NSW National Parks and Wildlife Aboriginal Sites Register was searched to determine if crayfish remains were widely reported from archaeological contexts. The regions specified in the computer search were based on the known distribution of Euastacus in eastern New South Wales, and focused on two broad regions — the N.S.W. south coast from Wollongong to the Victorian border, and the north coast from the Hunter Valley to the Queensland border (see Fig. 1). The search excluded sites immedi- ately adjacent to the coast, but included all open campsites, shelters with deposit, and middens, extending from the estuaries inland for approximately 200 km. The specified range included some Euastacus habitats associated with limestone, such as the Stroud and Dungog districts and west of Wauchope in the north as well as headwaters of the Shoalhaven and Moruya Rivers in the south. Following up on selected Site Register data 23 unpublished excavation reports were examined to see if crayfish remains were record- ed in the faunal species lists. RESULTS AND DISCUSSION Of over 1,500 site records examined in this survey, in no case was there a clear indication that freshwater crayfish remains were present. There were some references to ‘possible fragments of lobster shell’ and ‘crustacean shell, possibly lobster’; however, these were at sites close to the coast suggesting that the remains were probably those of marine crustaceans. The lack of crayfish remains is puzzling as this group of macro- invertebrates is the most abundant in many of the coastal aquatic systems, from middle reaches to headwaters, and larger species have robust skeletal components. Several aspects of this situation warrant further comment. Firstly, it was considered that preservation conditions in many of these areas may not be good due to light, acidic soils and consistently wet conditions in upland valleys; however, investigations of collections made in limestone areas (e.g. Armidale, Jenolan areas) have not yielded any further data. Other researchers also report a low incidence of crustacean remains at coastal sites (Attenbrow 1995, pers. comm.). In south-eastern Queensland the only recognisable crustacean remains at coastal sites, excavated to date, are the chelae (claws) of crabs (Robins 1996, pers. comm.); however, the chelae of the larger Euastacus would be equally robust and so could be expected to remain relatively intact. Secondly, the data now available about the biology of several larger Euastacus species (Honan and Mitchell 1995; Turvey and Merrick 1997a,b,c,d) indicate that these crays have life cycle strategies based on: slow growth and late maturation at a large size: low fecundity with annual (or less frequent) breeding; low adult mortality and individual longevity. When these features are considered, in conjunction with a territorial nature Proc. LINN. SOc. N.S.W., 119. 1998 104 LIMITED ABORIGINAL USAGE OF EUASTACUS and small home ranges, it is clear that local Euastacus populations could be quite effec- tively controlled (in terms of numbers of adults and breeding potential) by minimal culling (Barker 1990). Thirdly, even though the evidence suggests that highland areas were not as inten- sively populated or occupied as lower coastal areas, they were visited and used as transit routes (Bowdler 1981). Furthermore, native predators or scavengers would disturb remains of Aboriginal harvests. Dingoes or water rats would destroy remains, or render them unrecognisable, by chewing; whereas goannas, would disperse or destroy remains when digging for food (King and Green 1993). In summary, it 1s suggested that the paucity of crayfish in eastern Aboriginal sites is due to a combination of the following factors: (a) inadequate or incomplete sorting of existing collections; (b) archaeological sampling in areas where, coincidentally, few crayfishes were harvested; (c) poor preservation of crustacean remains due to unsuitable physicochemical conditions; (d) destruction and dismemberment of freshly discarded carcasses by native scavengers; (e) a low level of utilisation of eastern crayfish populations (resulting in a very low inci- dence of recognisable remains being preserved). Any conclusion relating to the level of Aboriginal exploitation of this dominant, edible, macro-invertebrate group is impossible, pending analyses of investigations of additional archaeological sites. In addition to robust pieces of chelae the other elements that are most likely to persist are gastric mill ossicles and the gastroliths. ACKNOWLEDGEMENTS Appreciation is expressed to Val Attenbrow, Roger Springthorpe (Australian Museum, Sydney), Richard Robins (Queensland Museum, Brisbane) as well as Mark Veerey, and other N.P.W.S. staff and col- leagues for their interest, comments and advice. We are grateful to Mr. J. Cleasby, School of Earth Sciences and Miss P. R. Davies, Graduate School of the Environment, Macquarie University for assistance with figure and manuscript preparation respectively. These studies were supported by a Macquarie University research grant. REFERENCES Barker, J. (1990). Spiny freshwater crayfish management strategy. Fisheries Management Report (34), 1-17. Department of Conservation & Environment, Melbourne. Bowdler, S. (1981). Hunters in the highlands: Aboriginal adaptations in the Eastern Australian Uplands. Archaeology in Oceania 16(2), 99-111. Campbell, I.-C. (1978). Settlers and Aborigines: the pattern of contact on the New England Tableland 1832-1860. In “Records of times past’ (Ed. I. McBryde), pp 5-16. (Australian Institute of Aboriginal Studies, Canberra). Flood, J. (1980). ‘The moth hunters. Aboriginal prehistory of the Australian Alps’. (Australian Institute of Aboriginal Studies, Canberra). Godfree, R. (1995). Analysis of vertebrate bone remains from an aboriginal shell midden located at Kurnell, N.S.W. B.Sc. (Hons) thesis, Macquarie University. Hall, J. (1986). Exploratory excavation of Bushrangers Cave (Site LA:A11), a 6000-year-old campsite in south- east Queensland: preliminary results. Australian Archaeology 22, 86-103. Honan, J.A. and Mitchell, B.D. (1995). Catch characteristics of the large freshwater crayfish, Euastacus bispinosus Clark (Decapoda: Parastacidae), and implications for management. Freshwater Crayfish 10, 57-69 Horwitz, P. and Knott, B. (1995). The distribution and spread of the yabby Cherax destructor complex in Australia: speculations, hypotheses and the need for research. Freshwater Crayfish 10, 81-91. Isaacs, J. (1987). ‘Bush food. Aboriginal food and herbal medicine’. (Ure Smith Press, Sydney). King, D. and Green, B. (1993). ‘Goanna. The biology of varanid lizards’. (New South Wales University Press, Sydney). Lourandos, H. and Ross, A. (1994). The great ‘intensification debate’: its history and place in Australian archaeology. Australian Archaeology (39), 54-63. Proc. LINN. SOC. N.S.W., 119. 1998 J.L. KOHEN AND J.R. MERRICK 105 Merrick, J.R. (1993). ‘Freshwater crayfishes of New South Wales’. (Linnean Society of New South Wales, Sydney). Merrick, J.R. (1995). Diversity, distribution and conservation of freshwater crayfishes in the eastern highlands of New South Wales. Proceedings of the Linnean Society of New South Wales 115, 247-258. Morgan, G.J. (1997). Freshwater crayfish of the genus Euastacus Clark (Decapoda: Parastacidae) from New South Wales, with a key to all species of the genus. Records of the Australian Museum, Supplement (23), 1-110. Owen, J.F. and Merrick, J.R. (1994a). Analysis of coastal middens in south-eastern Australia: sizing of fish remains in Holocene deposits. Journal of Archaeological Science 21, 3-10. Owen, J.F. and Merrick, J.R. (1994b). Analysis of coastal middens in south-eastern Australia: selectivity of angling and other fishing techniques related to Holocene deposits. Journal of Archaeological Science 21, 11-16. Turvey, P. and Merrick, J.R. (1997a). Reproductive biology of the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales 118, 133-157. Turvey, P. and Merrick, J.R. (1997b). Population structure of the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales 118, 159-176. Turvey, P. and Merrick, J.R. (1997c). Moult increments and frequency in the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales 118, 189-206. Turvey, P. and Merrick, J.R. (1997d). Growth with age in the freshwater crayfish, Euastacus spinifer (Decapoda: Parastacidae), from the Sydney region, Australia. Proceedings of the Linnean Society of New South Wales 118, 207-217. Versteegen, M. and Lawler, S. (1997). Population genetics of the Murray River crayfish Euastacus armatus. Freshwater Crayfish 11, 146-157. Proc. LINN. SOc. N.S.W., 119. 1998 se sai eater! ER ob pen ey een es ad aster ath eon He adi ht ite st vere = - oe ~ Wks — Set 3 ray b) ae tg 4)\ r ’ : : 4+ 7 » ~ é &- 1 ; : ¥ ae Ven Alt oir a : ; - . a u = a> Bahan pees Tha RATT cre oe Vikan 2 : a 4 7 ber = 3a } au : r Lie 5 ae ‘hy Als ; ; 7 : 7 eik, ic ais ¢ fay i Weeseye Ria neath 1 Oey a ae Ss 7 , al cor ny Vee rte ae owt a ae Meld, L k f : *. ; : io ase © avon ieee ep 10: (ile prin ike nit at Perret a) 647 Pr) = Pate vie cl Cyisieal 1. ee : Rte) dawn, (VS pk ty cet 7 7 > saend Sieg od eee Tie Cig Aan Wegbes ey ‘ a ‘ _— a an A pect rn - *»> i - Pag hil fh We ie Ws rok we _ rw) * ae A ee (Pl a cor a : ' ae we , - mM : a -_ ae a 4 Pleistocene Frogs from Near Cooma, New South Wales MICHAEL J. TYLER', ANGELA C. DAVIS? AND CRAIG R. WILLIAMS! ‘Department of Zoology, University of Adelaide, SA 5005; and *Department of Geology, Australian National University, Canberra, ACT 2600 TYLER, M.J., Davis, A.C. AND WILLIAMS, C.R. (1998). Pleistocene frogs from near Cooma, New South Wales. Proceedings of the Linnean Society of New South Wales 119, 107-113. Several frog limb fragments and six excellently preserved ilia were collected by one of us (A.C.D.) from the Jilliby Formation at the Bunyan Siding fossil locality, 8 km NNE of Cooma, New South Wales. The material forms part of the Bunyan Siding Fauna and has been dated by thermoluminescence dating and palaeomagnetic studies as Middle Pleistocene. The frogs represent two extant species: the hylid Litoria citropa (Duméril and Bibron) and the leptodactylid (myobatrachid) Limnodynastes peronii (Duméril and Bibron). Litoria citropa is known from eastern Victoria and south-eastern New South Wales, whereas Limnodynastes peronii has a more extensive geographic range extending from eastern Queensland in a continuous arc to the south-east of South Australia and includes Tasmania. This is the first fossil record of L. citropa, whereas L. peronii is known from the Holocene of Hunter Island in Bass Strait. Fossil frogs have been reported only once previous- ly from New South Wales. Manuscript received 17 February 1997, accepted for publication 20 August 1997. KEYWORDS: Pleistocene, frogs, ilia, scapulae, Limnodynastes, Litoria, New South Wales. INTRODUCTION It was not until 1973 that the first fossil frog was discovered in Australia (Tyler 1974). Taken at Lake Palankarinna in northern South Australia, it was from a deposit ini- tially considered to be Miocene, but subsequently argued to be of Oligocene age (Lindsay 1987). Because the first fossil was an ilium, its identification required a comparative study of the ilia of all extant Australian genera, so permitting its resolution as a new genus (Tyler 1976). Subsequent examination of other Cainozoic deposits resulted in 32 fossil Australian species being known by 1994 (Tyler 1994), whilst Tyler et al. (1996) added Litoria raniformis to bring the total to 33. Only one fossil frog species has been reported from New South Wales: Limnodynastes dumerilii from Lake Menindee (Tyler 1994). This finding is despite the fact that the extant fauna of the extreme south-east is rich, including representatives of seven genera: the hylid Litoria Tschudi and the leptodactylids (myobatrachids) Limnodynastes Fitzinger, Crinia Tschudi, Mixophyes Giinther, Neobatrachus Peters, Pseudophryne Fitzinger and Uperoleia Gray. Here we report the discovery by one of us (A.C.D.) of the second known fossil frog fauna from New South Wales. Several axial and limb fragments, including six frog ilia, were found in the northern Monaro Region. The fragments form part of the Bunyan Siding Fauna and are of Middle Pleistocene age (Davis 1996). Proc. LINN. Soc. N.S.W., 119. 1998 108 PLEISTOCENE FROGS FROM COOMA eae FLAT NUMMERALLA - Soe ete" RIVER -¢ —N'Rose Brook’ ¢ i , a (Cloyne ¢ Vie BUNYAN SIDING SITE B® Bunyan oe | Railway NUMMERALLA Seg 5 km Figure 1. Locality map showing Bunyan Siding fossil site near Cooma, New South Wales. LOCALITY AND STRATIGRAPHY The frog specimens were found at the Bunyan Siding fossil locality, eight km NNE of Cooma (grid ref. 955961 on Cooma 1:100,000 sheet), south-eastern New South Wales (Fig. 1). The locality was first described by Ride et al. (1989) as a perched alluvial terrace deposit dissected by the present Rose Valley road. The site comprises two main Quaternary sedimentary units, the Jilliby Formation and the Nestle Brae Formation (Davis 1996). The Jilliby Formation is at the base of the Quaternary sedimentary sequence and disconformably overlies lake clay facies of the Early Miocene Bunyan Formation defined by Taylor and Walker (1986). The unit comprises some 20 horizontal beds in a generally fining upward gravel/sand/clay sequence of 2 m thickness (Fig. 2). The basal bed, a coarse gravel, extends the length of the site (over 150 m) while overlying sand and clay beds lens out with lateral facies changes. The upper contact of the Jilliby Formation is an irregular erosional surface marked in places by a cemented calcrete horizon and infilled by sediments of the overlying Nestle Brae Formation, a reddish coloured sandy unit up to 1.5 m thick interbedded with small gravel lag deposits, with a well-developed red earth soil in the upper 40 cm (Davis 1996). The Jilliby Formation facies sequence is interpreted as a large stream migrating across the valley, intermittently accreting channel, backswamp and flood plain facies, followed by a long period of surface stability with calcrete development and erosion (Davis 1996). Proc. LINN. SOC. N.S.W., 119. 1998 M.J. TYLER, A.C. DAVIS AND C.R. WILLIAMS 109 Re) Depth = 5 Mm) fi c ae 5 8 @ Sed. Dating JilibyFm 82 § & struct. sample Nestle Brae Formation 6 : c Onno G Cc Jilliby Formation Cc FROG BONES 50. rm mn 100 ) j \ ry 150 Tr Jilliby Formation PUR ETA 5418 a) 200 Bunyan Formation GRAIN SIZE SEDIMENTARY TYPE OF DATING STRUCTURE Ml clay 224 fine sand c calcrete thermoluminescence coarse sand \ bioturbation palaeomagnetics gravel ’™ cross bedding Figure 2. Stratigraphic log at Trench 11S, Bunyan Siding fossil site, near Cooma, New South Wales. Proc. LINN. Soc. N.S.W., 119. 1998 110 PLEISTOCENE FROGS FROM COOMA The majority of the Bunyan Siding Fauna and all of the frog material described in this paper were collected from the Jilliby Formation from a series of trenches excavated along the road cutting exposure. Three of the frog specimens (AM F98392-94) were col- lected in Trench 11S near the top of the unit in fine sandy clay facies (Fig. 2), while the remaining frog specimens were collected from Trench 11—12.5S (bed uncertain). AGE AND ASSOCIATED FAUNA A recent revision of the age of the locality (Davis 1996) indicates a Middle Pleistocene age for both the Jilliby and Nestle Brae Formations, not Late Pleistocene as previously thought (Ride et al. 1989). Thermoluminescence dating at the base of the Nestle Brae Formation and at the top of the Jilliby Formation (Fig. 2) indicate an age older than 100 ka, while palaeomagnetic samples from throughout the Jilliby Formation indicate Normal polarity implying an age within the Brunhes geomagnetic polarity stage (i.e. younger than 780 ka). Thus, the Bunyan Siding Fauna is interpreted as being of Middle Pleistocene age, but the position within the Middle Pleistocene is uncertain. The Bunyan Siding Fauna comprises material from both stratigraphic units, full descriptions of which are provided in Davis (1996). In addition to the frog material described here, associated taxa include eighteen species of mammal, one species of freshwater fish and three species of reptile. MATERIALS AND METHODS The specimens reported here have been deposited in the palaeontological collec- tion of the Australian Museum, Sydney. The identifications are based upon comparative studies of ilia in the osteological collection held at the Department of Zoology at the University of Adelaide. Descriptive terminology follows Tyler (1976, 1989). Scanning electron micrographs were produced using a Joel 6400 SEM at the Electron Microscopy Unit (R.S.E.S.), Australian National University. SYSTEMATICS ORDER: Anura FAMILY: Hylidae SUBFAMILY: Pelodryadinae Litoria citropa (Duméril and Bibron) Material Four ilia — AM F98391-94 Description None of the specimens is complete but there is sufficient evidence to confirm their conspecificity and specific identity. Each of the specimens is illustrated in Fig. 3. The dorsal prominence and dorsal protuberance of L. citropa are unusual, in being clearly demarcated by a narrow indentation surrounding the posterior and inferior bound- aries of the protuberance. The dorsal acetabular expansion (DAE) superiorly is on a level with or slightly above the superior margin of the ilial shaft. Proc. LINN. SOC. N.S.W., 119. 1998 M.J. TYLER, A.C. DAVIS AND C.R. WILLIAMS 111 Figure 3. Fossil ilia of Litoria citropa. A. Australian Museum 98392, B. 98394, C. 98391, D. 98393. Scale bar = 1 mm. The acetabulum is large and has a narrow, and clearly defined, circummarginal rim. The superior margin of the acetabular fossa extends slightly above the ventral mar- gin of the ilial shaft. The preacetabular zone is broad and gently curved. The subacetabu- lar zone is missing from all specimens. The ilial shaft is incomplete in all specimens. FAMILY: Leptodactylidae' SUBFAMILY: Limnodynastinae Limnodynastes peronii (Duméril and Bibron) Material Two ilia — AM F98395—96 Description All Limnodynastes species have an extremely large dorsal prominence and dorsal protuberance. In the case of L. peronii collectively these structures are huge, extending superiorly high above the dorsal rim of the ilial shaft (Fig. 4). The dorsal acetabular expansion (DAE) is high but the base of this structure has coalesced with the dorsal prominence, and thus it appears small and even vestigial com- pared with congeners. Nevertheless, as in other Limnodynastes species, the anterior mar- gin of the DAE rises steeply. The acetabulum is relatively large, and the superior rim of the acetabular fossa extends above the ventral margin of the ilial shaft. The pre-acetabular zone is narrow, and the sub-acetabular zone is dilated into a spatulate form of which the inferior margin is incomplete. Proc. LINN. Soc. N.S.W., 119. 1998 112 PLEISTOCENE FROGS FROM COOMA Figure 4. Fossil ilia of Limnodynastes peronii. A. Australian Museum 98396, B. 98395. Scale bar = 1 mm. The distal end of the ilial shaft of AM F98395 is irregular, consistent with the interpretation that the shaft is complete, being replaced with cartilage at the position of its articulation with the sacrum. The total length of this specimen is 14 mm. The other representative (AM F98396) is a left illum of a different individual with an incomplete ilial shaft and is also 14 mm long. DISCUSSION The geographic distribution of L. citropa has recently been revised by Anstis and Littlejohn (1996), and reveals an apparent gap of approximately 200 km northwards from the New South Wales-Victoria coastal border. The fossil site reported here is just within the northern portion of that gap, so bridging it slightly. In the case of Limnodynastes peronii the site is well within the current geographic range of that species. REFERENCES Anstis, M. and Littlejohn, M.J. (1996). The breeding biology of Litoria subglandulosa and L. citropa (Anura: Hylidae), and a re-evaluation of their geographic distribution. Transactions of the Royal Society of South Australia 120, 83-99. Davis, A.C. (1996). Quaternary mammal faunas and their stratigraphy in the northern Monaro region, south- eastern Australia. PhD thesis, Geology Department, Australian National University, Canberra. Proc. LINN. SOC. N.S.W., 119. 1998 M.J. TYLER, A.C. DAVIS AND C.R. WILLIAMS 113 Lindsay, J.M. (1987). Age and Habitat of a monospecific foraminiferal fauna from near-type Etadunna Formation, Lake Palankarinna, Lake Eyre Basin. South Australian Department of Mines and Energy Report 87/93, unpublished. Ride, W.D.L., Taylor, G., Walker, P.H. and Davis, A.C. (1989). Zoological history of the Australian Alps - The mammal fossil-bearing deposits of the Monaro. In * The Scientific Significance of the Australian Alps’ (Ed. R. Good) pp. 79-110. (Australian Alps Liaison Committee: Canberra). Taylor, G. and Walker, P.H. (1986). Tertiary Lake Bunyan, Northern Monaro, NSW, Part 11: facies analysis and palaeoenvironmental implications. Australian Journal of Earth Sciences, 33: 231-251. Tyler, M.J. (1974). First fossil frogs from Australia. Nature 248, 711-712 Tyler, M.J. (1976). Comparative osteology of the pelvic girdle of Australian frogs and description of a new fos- sil genus. Transactions of the Royal Society of South Australia 100, 3-14. Tyler, M.J. (1989). A new species of Lechriodus (Anura: Leptodactylidae) from the Tertiary of Queensland, with a redefinition of the ilial characteristics of the genus. Transactions of the Royal Society of South Australia, 113, 15-21. Tyler, M.J. (1994). ‘Australian frogs. A natural history’. Reed, Sydney. Tyler, M.J., Barrie, D.J. and Walkley, R.W. (1996). First fossil record of the hylid frog Litoria raniformis (Keferstein). Transactions of the Royal Society of South Australia 120 (2), 69. ENDNOTE 'We persist with the use of the family name Leptodactylidae because of evidence of the lack of a single synapomorphy uniting the sub-families Myobatrachinae and Limnodynastinae within what is currently regarded as the Myobatrachidae. The inference is that further study will result in Limnodynastes and other limnodynastines being referred to a separate family. Accordingly, it is expedient to maintain conservatism of nomenclature rather than to include Limnodynastes in the Myobatrachidae, to which it evidently does not belong. Proc. LINN. SOc. N.S.W., 119. 1998 yori ® . ve i ae (0@0 i (A) avi hi dew aft dtea & ani , i] | ned 7 Dieses qpavg' nf Ty, a Huy ee ee Tee a tw Let go ii P ‘i ; 4 4 iv 2 , ae [x= & : we la ; - a” aie ae a “ ; A tueiei et eb a . a Tertiary Climatic Evolution and the Development of Aridity in Australia HELENE A. MARTIN School of Biological Science, University of New South Wales, Sydney NSW 2052 Martin, H.A. (1998). Tertiary climatic evolution and the development of aridity in Australia. Proceedings of the Linnean Society of New South Wales 119, 115-136 The climatic record is deduced from palaeobotany and some other relevant studies. The record from southeastern Australia is the most comprehensive and is presented for com- parison with other parts of Australia where there are few studies. In the southeast, the precipi- tation was well above the critical level for rainforest (1500 mm) during the Palaeogene. In the mid-late Miocene, precipitation declined to less than 1500 mm and in the late Pliocene- Pleistocene, there was a further decrease to about 500-800 mm. In the northeast of Australia, the precipitation remained well above the critical level for rainforest throughout the Tertiary and most of the Quaternary, hence this region is a refuge for many rainforest taxa. When the northwest of Australia is compared with the southeast, the climate of the former was drier than the latter throughout the late Tertiary and Quaternary. Grasses were rare until the late Miocene when they show a steady increase which is maintained through the remainder of the Cainozoic. Grasslands developed first in the north- west and, presumably, Central Australia, but development was later in the southeast. A trend towards aridity started in the mid Miocene and continued through the late Tertiary. A degree of aridity was reached about the late Pliocene-early Pleistocene, but aridity has intensified during the Quaternary, especially in the latter part. Manuscript received 20 May 1997, accepted for publication 19 November 1997. KEY WORDS: Aridification, Australia, grasslands, palaeoclimate, Tertiary, vegetation history. INTRODUCTION Weather and climate have a daily fascination. The extremes cause discomfort that lead some to claim ‘the climate is changing’. While it is difficult to distinguish short term variation from a trend or change on the basis of these small scale observations, it is clear that climate has changed, and has changed drastically in the past: the Australian deserts once supported luxuriant vegetation. This study reconstructs the Tertiary climate from the late Eocene, some 40 million years ago, when Antarctica was mostly ice free and Australia was almost entirely forest- ed, and covers the crucial development of aridity. By the end of the Tertiary, some 2—3 million years ago, the climate resembled the modern status, but aridity has intensified, especially in the last half million years. The history of the vegetation is a major source of evidence of climatic change, but there are other studies which complement this history and add invaluable insights into past environments. These past changes suggest that the climate is likely to change in the future, and it could change dramatically from entirely natural causes. The history of the vegetation is constructed from fossil evidence, mainly from palynology, and to a lesser extent, from macrofossils. Each source has both strengths and limitations. Pollen may be extracted from bore samples, hence palynology accesses sedi- ment which may be hundreds of metres deep and the record from this source is compre- hensive. Fossil leaves, the major source of macrofossil data, however, must be recovered from surface outcrops, although cuticles may be found in bore samples. Both pollen and Proc. LINN. SOc. N.S.W., 119. 1998 116 TERTIARY CLIMATE IN AUSTRALIA leaves may show affinities with living taxa, but identification with a modern species is not always possible. Affinities are usually made with genera or families, but a fossil pop- ulation may not coincide with any living taxon, even when natural affinities are evident. Dust whipped up by wind may be blown out to sea and deposited in marine sedi- ments. The dust fluxes in deep sea cores record the intensity of deflation on land (Glasby 1971; McTainsh 1989; Hesse 1994). Phytoliths, the silica bodies produced by grasses and many other plants, may form part of the dust flux and are another source of evi- dence. Palaeotemperatures deduced from oxygen isotope analyses of deep sea cores (Shackleton and Kennett 1975; Savin et al. 1975), corrected for continental ice volume where necessary (Feary et al. 1991; Isern et al. 1996), indicate surface sea temperatures and the trends may be extrapolated to the land. These records are independent of palaeobotany and hence are invaluable for confirmation of the record from palaeobotany, as well as filling in gaps in the record. Reconstruction of the vegetation is achieved from the floristics of the palynofloras. The ecological tolerances and climatic limitations of present day taxa and vegetation are applied to the fossil assemblage to deduce past climates. Leaf physiognomic characters, such as dimensions, the nature of the margin, drip tips etc. are a direct expression of cli- mate, irrespective of taxonomic affinities. The nature of the sediments also contains a cli- matic signal that is used as supplementary evidence. Numerous experimental studies show that most of the pollen in an assemblage at any one site has been produced locally, within a radius of half a kilometre or less. A little pollen may have been transported in from long distances. (For reviews of the numerous studies on this subject, see Birks and Birks 1980, and Martin 1993.) The pollen assemblage is a reflec- tion of the dominant type of vegetation and small, isolated patches of a different kind, such as may be found in sheltered gullies and gorges, may be palynologically invisible (Ladd 1979). Pollen in ocean cores, however, has been transported from land either by wind or runoff from rivers and studies of marine surface sediments show that the pollen in them is a general reflection of the regional vegetation on land (Mudie 1982; Turon 1984; Prell and van Campo 1986; Heusser 1988; Mudie and McCarthy 1994). All kinds of plant parts require the anaerobic conditions of lakes, swamps, bogs etc. for preservation. The vegetation growing around these sites, which occupy the low- est part of the topography, are best represented in the fossil record. Plants growing on sites distant from the sedimentary basin, such as the hilltops or steep slopes usually have little chance of being fossilised. The vegetation units identifiable in the fossil record are usually fairly broad and general. The units of vegetation used in this study are those in common usage and are defined thus: Rainforests Rainforests or closed forests (Specht 1970) are usually structurally and floristically complex. They are found in the better watered environments. Eucalyptus is not a normal part of rainforests. There are many different types of rainforest in tropical, subtropical and temperate regions. In Australia, tropical rainforests are best developed north of 21° latitude, although patches may extend further south. Subtropical rainforests are most common between 21° and 35° south and temperate rainforests are the dominant type south of 35° (Webb 1959), although small patches of the latter may be found as far north as 28°. Nothofagus is found in temperate rainforests which are especially well developed in Tasmania, New Zealand and Chile, extending to about 50—55°S (Riley and Young 1972). Rainforests may be divided into ‘wet’, with Nothofagus, and ‘dry’, usually with various gymnosperms (Kershaw et al. 1994). In drier locations, some scrubs and vine thickets are closely related floristically to rainforests and hence are included in this clas- sification (Webb 1959; Webb and Tracey 1981). Proc. LINN. SOc. N.S.W., 119. 1998 H.A. MARTIN 117 Sclerophyll forests or open forests Sclerophyll or open forests (Specht 1970) are most commonly dominated by Eucalyptus and/or Casuarina/Allocasuarina. Wet sclerophyll forests have rainforest or other mesic taxa in the understorey layers, and if left unburnt and undisturbed, may revert to rainforest (Ashton 1981). Dry sclerophyll forests do not have rainforest taxa. The understorey in these forests contains sclerophyllous shrubs (Gill 1981). Fire is an integral part of the environment of sclerophyll vegetation (Ashton 1981; Gill 1981). Open vegetation In forests, the tree canopy forms an almost continuous layer whereas in woodlands, the trees are well spaced. If there are few trees, the vegetation is more open and there is a well developed layer of shrubs, grasses and/or herbs. With a dense tree cover, insufficient light filters through the trees to support the ground layer (Specht 1970). In this study, open vegetation refers mainly to shrubland, grassland, and/or herbfield. The sites studied for Tertiary palynology are shown in Fig. 1. Southeastern Australia has been studied most intensively but most of these studies present a disjointed sequence which must be pieced together for a coherent history. Dating is usually achieved by correlation with a dated sequence as most sediments cannot be dated directly and there is little independent evidence of the age. There are a few sites on the continen- tal shelf and in the deep sea that yield pollen which must have come from the vegetation on land. These sites present a more continuous sequence that is well dated by indepen- dent evidence from marine foraminifera. Figure 1 shows that there are large areas for which there is no evidence. This review presents the climatic evolution of southern Australia, which has a rela- tively good record, for comparison with northern Australia where the evidence is sparse. SOUTHERN AUSTRALIA The climate over most of southern Australia is temperate, becoming subtropical in the north and dry continental, semi-arid and arid in the central and inland regions (Foley 1954). Patches of rainforest are found along the east coast and in Tasmania but Eucalyptus forests and woodlands are dominant over most of the area. Open shrublands and grasslands are found in the semi-arid and arid regions. Most of the evidence comes from southeastern Australia and this region covers Tasmania, Victoria, New South Wales and the southeast of South Australia. There are a few sites in central and southwest Australia. Palaeovegetation In southeastern Australia, the Lachlan River Valley on the western Slopes of the Great Dividing Range (H-F-C on Fig. 1) has an almost continuous record from late Eocene to the Pleistocene (Fig. 2). The vegetation was rainforest with abundant Nothofagus throughout the Oligocene, arguably the wettest period in the Tertiary. In the late Oligocene-early Miocene, Nothofagus declined, but the vegetation was still predomi- nantly rainforest (Martin 1987). The mid Miocene was a time of profound change when rainforests were decimated and myrtaceous forests became predominant. The carbonised particle or charcoal content is greater in the myrtaceous pollen assemblages than in the rainforest assemblages, show- ing that periodic burning had become part of the environment, thus inferring that the veg- etation was sclerophyll forests and not rainforest, for the latter rarely burns. This indirect Proc. LINN. Soc. N.S.W., 119. 1998 118 TERTIARY CLIMATE IN AUSTRALIA Melville I ‘Riversleigh Lake Eyre ,@ apa Tee Lake romee @ Sites studied for Tertiary palynology @) Deep sea sites studied for palynology Figure I. Sites studied for Tertiary palynology. The sites used for reconstructions in Fig. 2 are: H, Hillston; F, Forbes; C, Cowra, all of the Lachlan River Valley; A, the Aquarius well on the continental shelf; D, Darling River; N, Northern New South Wales; W, western region of southeastern Australia. method of identifying eucalypt vegetation is necessary as Eucalyptus pollen is difficult to distinguish from other myrtaceous pollen types (Martin 1987). Some rainforest taxa are present in these myrtaceous assemblages, hence the interpretation of wet sclerophyll forests. There is a hiatus of non deposition and/or erosion in the late Miocene, thought to correspond with the late Miocene low sea level (Fig. 2). In the late Miocene-early Pliocene, there is a brief resurgence of rainforest, followed by a return to wet sclerophyll forest. In the late Pliocene, the tree taxa declined and the vegetation became more open, viz, woodlands, grasslands and herbfields (Martin 1987). When the records from other sites in southeastern Australia are compared with that of the Lachlan River Valley, the trends are generally similar but with some local differ- ences. In Tasmania, the palynological record is limited (Kershaw et al. 1994), but there is a rich macrofossil record (Hill 1992; Carpenter et al. 1994). The Tertiary vegetation con- tained abundant rainforest taxa which persisted into the Quaternary when taxa common in modern sclerophyllous heathlands and woodlands became increasingly common. The late Quaternary vegetation was mainly Eucalyptus, Asteraceae and Poaceae, but Nothofagus and other rainforest taxa were still present as well. Thus rainforest has had a Proc. LINN. SOC. N.S.W., 119. 1998 H.A. MARTIN 119 metres GLOBAL SEA LEVELS q Ww uj oc 7) LW & Or . Snr Lo cs > i OE Oxygen isotope temperature, °C PRECIPITATION mm/pa Marked seaonal dry period Relatively high humidity, year round Present day Rainforest HILLSTON charcoal particles, Rainforest VEGETATION Wet sclerophyll forest Wet sclerophyll forest Abundant Myrtaceae and some rainforest species COWRA-FORBES Less Nothofagus Abundant Nothofagus y ANS00Md SANS00IN FNS90059I10 3AN39004 (2) - 20 o ° oO Tv sieak uoll|l\ Figure 2. Reconstructions of the vegetation and climate from late Eocene into Pleistocene for the Lachlan River Valley, from Hillston to Cowra. See Fig. | for location. Present day levels of precipitation are for: H, Hillston; F, Forbes; C, Cowra. Oxygen isotope temperatures are from Shackleton and Kennett (1975) and global sea levels from Haq et al. (1987). Note: The surface sea temperatures are not corrected for continental ice volume (see text). Proc. LINN. Soc. N.S.W., 119. 1998 120 TERTIARY CLIMATE IN AUSTRALIA continuous presence through the Neogene and Quaternary in Tasmania (Carpenter et al. 1994; Kershaw et al. 1994). The early—mid Miocene of the Latrobe Valley, southeast Victoria (Fig. 1) has abun- dant Nothofagus and other rainforest taxa. Casuarinaceae and sclerophyllous taxa are present as well. The rainforest taxa are also well represented in the late Miocene but have disappeared in the late Pliocene-early Pleistocene, when Eucalyptus, Asteraceae and Poaceae become abundant (Kershaw et al. 1994; Blackburn and Sluiter 1994), In the Southern Highlands (Lake George and other sites in the region, Fig. 1), rain- forest is well represented in the early-mid Miocene and extends into the late Pliocene, after which Asteraceae and Poaceae become prominent (Kershaw et al. 1994). In north- ern New South Wales (N on Fig. 1), rainforest is well represented in the mid-late Miocene. In the Pliocene, there is very little Nothofagus or ‘wet’ rainforest, but the gym- nosperm taxa, and especially Araucariaceae (‘dry’ rainforest) are well represented. Asteraceae and Poaceae become abundant in the Pliocene-Pleistocene (Kershaw et al. 1994). The western region of southeast Australia (W on Fig. 1) shows similar patterns, with rainforest well represented in the late Oligocene to early-mid Miocene and dimin- ishing in the Pliocene (Kershaw et al. 1994). Late Oligocene—early Miocene assemblages along the Darling River, the most northwesterly part of southeastern Australia (D on Fig. 1), have a diverse rainforest flora, but in low frequencies and the vegetation was mainly sclerophyll, with some pockets of rainforest, probably along the river valleys, the habitats with the most favourable mois- ture relationships. Asteraceae and Poaceae become common in the Pliocene-Pleistocene (Martin 1997). The sites studied from southwest Australia and southern coastal South Australia (Fig. 1) are mainly Eocene in age, with one Pliocene site. The Eocene palynofloras have abun- dant Nothofagus and other rainforest taxa (Macphail et al. 1994) and are generally similar to those in southeastern Australia. By the Pliocene, the vegetation had become sclerophyl- lous forests or woodlands dominated by Casuarinaceae or Myrtaceae (Bint 1981). Palaeoclimate The precipitation deduced from the climatic requirements of comparable present- day vegetation (see Martin 1987) is shown in Fig. 2. The early Oligocene was a time of very high precipitation, over 1800 mm, with high and constant humidities. Precipitation declined somewhat in the Oligocene, and in the early Miocene, it was probably closer to 1500 mm, the lower limit for widespread rainforest in New South Wales today. In the mid-late Miocene, precipitation decreased to 1000-1500 mm, the limits for wet sclero- phyll, and there must have been a marked dry season to allow burning on a regular basis. In the late Miocene-early Pliocene, precipitation increased to over 1500 mm at the time of the rainforest revival. Subsequently, it decreased to below 1500 mm and in the late Pliocene-Pleistocene, to about 500-800 mm (Fig. 2). The present day levels of precipita- tion are shown on Fig. 2. for comparison, and they are less than those for the late Pliocene-early Pleistocene. When the palaeovegetation of the other sites in southeastern Australia are com- pared with that of the Lachlan River Valley, they suggest that it was wetter in Tasmania and the southern highlands and drier in the northwest of the region, just as it is today (Martin 1986; 1990a). Temperatures may be deduced from the palaeovegetation in a similar way to that of precipitation, but these two factors are not independent. When temperatures are higher, there is more evaporation and the climate is more humid. Once the climate becomes colder, it is also drier. The oxygen isotope record from deep sea cores (Shackleton and Kennett 1975) has been used to estimate surface sea temperature but it also indicates continental ice volume. In the Neogene and Quaternary, however, the record is influ- Proc. LINN. SOC. N.S.W., 119. 1998 H.A. MARTIN 12] enced by both factors and corrections for ice volume are applied (Feary et al. 1991; Isern et al. 1996). Figure 2 presents the surface sea temperatures from Shackleton and Kennett (1975). Temperatures are high in the late Eocene, probably the highest for the Tertiary, decreasing in the Oligocene, when the Antarctic ice cap was developing, and increasing in the early-mid Miocene. Temperatures show a marked decline in the late Miocene, with a subsequent rise in the late Miocene-early Pliocene, followed by a sharp decline in the late Pliocene. Temperatures of the Pleistocene fluctuated with the glacial/interglacial cycles. The decline in temperatures through the Tertiary parallels the build up of ice on Antarctica (Kennett 1993). Precipitation and temperatures (Fig. 2) are roughly parallel, especially from the Miocene on. Warmer times were also wetter and conversely, colder times are drier, as expected. In the Australian context, precipitation has been the overriding control, hence a clear, unambiguous temperature signal is not evident in the record from the palaeovegetation. The precipitation record in Fig. 2 suggests that there are long periods of relative climatic stability and short periods of dramatic change. These periods of change coincide with some worldwide events which have a profound impact on climate. Global changes in sea levels (Haq et al. 1987) are also shown on Fig. 2. When sea levels are high, the continental shelves and low lying areas on land are flooded. These shallow seas warm up and evaporation is high, hence precipitation is increased. When sea levels are low, the continental shelves are exposed, the land is well-drained and the deep seas bordering the shelves are cold, hence evaporation and precipitation are both low. The record of sea level, temperatures and precipitation show roughly similar trends from the Miocene on. There are high levels in the mid Miocene, low levels in the late Miocene, with another high in the early Pliocene. The peaks do not coincide exactly, for there may be an apparent lag due to some specific local factor or imprecise dating. The parallel trends do not hold as well in the latest Tertiary because the ice volume on Antarctica had become a significant factor. The temperature curve from Shackleton and Kennett (1975) was selected because it is the record closest to Australia (see Fig. 2) and hence has most relevance to southeastern Australia, but this curve is essentially similar to that of Savin et al. (1975) for the Atlantic and Indian Oceans, and reflects general global trends in climate. In these global records of sea levels and oxygen isotope surface sea temperatures, the periods that stand out are as follows: 1. High sea levels and warmer surface sea temperatures in the mid Miocene and the late Miocene-early Pliocene. 2. Low sea levels and cooler temperatures in the mid-late Miocene and again in the late Pliocene-early Pleistocene. The Quaternary glacial/interglacial cycles commenced in this latter period. These two periods mark substantial decreases in both temperature and precipitation. The times of marked change in the palaeobotanical record coincide with these glob- al changes in sea level and temperatures. The periods of low sea levels and decreased tem- peratures, with accompanying lower precipitation, were devastating to the rainforest. NORTHERN AUSTRALIA The northern part of Australia is tropical and the coastal strip subequatorial (Foley 1954) and experiences summer monsoonal rains (Leeper 1970). Rainforest is found in patches along the north and east coast and is best developed in northeast Queensland, but overall, rainforest occupies only a relatively small area. Most of the region is covered with open forest, woodland and/or shrubland (Specht 1970). Proc. LINN. Soc. N.S.W., 119. 1998 122 TERTIARY CLIMATE IN AUSTRALIA Northeast Queensland Deep sea sites off Cairns and Mackay respectively (Fig. 1) have good records of late Miocene-Pleistocene palynofloras. The vegetation on the land nearest both sites was predominantly araucarian rainforest and casuarinaceous forest (Martin and McMinn 1993). The casuarinaceous pollen could indicate either Gymnostoma, a rainfor- est taxon, or Casuarina/Allocasuarina, sclerophyllous taxa (Kershaw 1970). There is, however, a suite of other sclerophyllous taxa, hence there must have been some sclero- phyll vegetation. There is some cyclical variation between the araucarian and casuarina- ceous vegetation, but no marked change. The pattern of change seen in southeastern Australia is not evident here, and extensive rainforest continues into the Pleistocene, until 120,000 years ago when the araucarian rainforest declines (Kershaw et al. 1993). Today, the Cairns district is the wettest region of Australia, and it may be that in the Neogene, the rainfall was well above the lower limits for rainforest, such that fluctua- tions did not provoke the decline in rainforest seen elsewhere in Australia. These deep sea sites collect pollen principally from the coastal vegetation which would have been closest to the site, whether the sea level was high or low. In contrast, a site on land would register the migration of the vegetation zones as they moved in unison with sea levels (Martin and McMinn 1993). A site on the Atherton Tablelands, inland from Cairns, is thought to be Pliocene- Pleistocene in age (Kershaw and Sluiter 1982). Podocarpus, Nothofagus, Casuarinaceae and Myrtaceae were at times prominent in the vegetation. Eucalyptus is not separated from rainforest Myrtaceae, and there is a wealth of low frequency rainforest taxa. Araucarians were minimal (Kershaw and Sluiter 1982). This assemblage from the table- lands reflects different vegetation to that on the coast, but it is essentially rainforest. The A.O.G. Aquarius No. | well on the continental shelf (A on Fig. 1), has a good Neogene sequence (Hekel 1972). Araucarians, Casuarinaceae and Myrtaceae were common in the vegetation, the latter in contrast to the deep sea sites where Myrtaceae is minimal. Rainforest was a considerable part of the vegetation here also. Rainforest, and hence a relatively high precipitation, was maintained throughout the Neogene in northeast Australia when rainfall was decreasing over most of the conti- nent. Environments favourable for rainforest have continued to the present day and the Cairns region is still the wettest part of Australia today (Leeper 1970) and has the most extensive tracts of rainforest. Feary et al. (1991) have reconstructed the changes in palaeotemperatures using sur- face sea temperature derived from the Tertiary oxygen isotope record corrected for ice volume and the decreasing latitude with continental drift (Fig. 3). For the Great Barrier Reef, temperatures were subtropical in the Eocene, temperate in the Oligocene, subtropi- cal in the Miocene, and finally becoming tropical in the Pliocene-Pleistocene when the Great Barrier Reef entered tropical latitudes (see Fig. 3) (Feary et al. 1991). On average, temperatures were above the minimum required for tropical reef growth (20°C) from the middle Miocene to Holocene, except for intervals in the late Miocene-early Pliocene, when the temperatures fell to between 18° and 20°C repeatedly (Isern et al. 1996) Northwest Australia There are three Tertiary palynological sites in northwest Australia (Fig. 1). The two on land are ?Eocene in age (Truswell and Harris 1982) and contain some rainforest taxa found in deposits of a similar age in southeastern Australia. There is also an almost con- tinuous sequence from late Miocene to the Recent in a deep sea site, off Port Hedland. There, the pollen would have originated from the land to the south, from Port Hedland and southwards, as there is a north-south canyon down the edge of the shelf which acted as a funnel (Martin and McMinn 1994). In the late Miocene the vegetation was casuarinaceous forests and there were no Proc. LINN. SOC. N.S.W., 119. 1998 H.A. MARTIN 123 25 TROPICAL 20R-“A- Ho ooo oe KH eo \wee—- H- Mee Heke em me eK Ke Ke TEMPERATE 10 Estimated surface sea temperature (° C) on 5 e]Plig Miocene Oligocene 0 20 40 60 Time (million years) Figure 3. Inferred surface sea temperature for northeast Australia (from Feary et al. 1996). The upper curve represents the northern end of the Great Barrier Reef and the lower curve, the southern end. For further expla- nation, see text. unequivocal rainforest taxa. The spores of ferns, bryophytes etc., indicative of damp habitats, are minimal, hence the climate was relatively dry. The casuarinaceous forests decline and are replaced by grasslands. Acacia is present with relatively high frequen- cies (for Acacia) which is always under-represented, suggesting that it was an impor- tant taxon in the vegetation. The chenopod type, pollen of Chenopodiaceae, is indica- tive of arid vegetation and increases in the Pliocene and Pleistocene. There is the mini- mal Myrtaceae pollen, hence minimal Eucalyptus and the lack of eucalypt dominated vegetation. The vegetation of the region today has large tracts where Acacia shrub- lands and hummock-tussock grasslands are dominant. This deep sea site probably reflects the development of the hummock-tussock grasslands (Martin and McMinn 1994; Specht 1970). A remarkable macrofossil assemblage from Melville Island (Fig. 1) is, unfortunate- ly, undatable. Cupressaceae, Grevillea, several other taxa of Proteaceae and Melaleuca indicate a non-rainforest community which probably had a seasonal climate (Pole and Bowman 1996). When compared with southeastern Australia, the lack of rainforest and earlier increase in grasses suggests that it was drier in the northwest. Today, the northwest is much drier than the southeast, hence in general terms, climatic gradients in the Tertiary were parallel to those of today. Proc. LINN. SOc. N.S.W., 119. 1998 124 TERTIARY CLIMATE IN AUSTRALIA CENTRAL AUSTRALIA Most of the sites in Central Australia, around Lake Eyre and in the Northern Territory are late Paleocene-late Eocene in age. The vegetation was mainly forests and herbaceous swamps. The rainforests were richly diverse in gymnosperms and there was some Nothofagus, but the latter was not as abundant as in coastal regions. There was a rich angiosperm flora also, and proteaceous taxa were prominent (Wopfner et al. 1974; Truswell and Harris 1982; Alley 1985; Sluiter 1991). Fossil leaf floras with small-sized leaves and an absence of drip-tips indicate some sclerophyllous vegetation (Greenwood et al. 1990, Christophel et al. 1992). Silicified moulds and casts of the fruits of Eucalyptus; Angophora, Leptospermum, Melaleuca/Callistemon and Calothamnus (Lange 1978) add to the sclerophyllous element. There was a mosaic with rainforest in the wetter habitats on the floodplains and along the watercourses with the sclerophyll vegetation on the drier and more nutrient deficient interfluves. Palaeobotanical material younger than the late Eocene is rare in Central Australia. Once the climate became drier, the swamps etc, required for preservation, became scarce. When pollen is deposited in a swamp, it must be buried deep enough to escape the destructive effects of a fluctuating water table. The deep weathering, so common in inland Australia, has undoubtedly destroyed much of the palaeobotanical evidence. A late Oligocene-early Miocene assemblage from Lake Frome contains rainforest gym- nosperms, a little Nothofagus and abundant swamp taxa. Grass pollen is rare (Martin 1990b). (Extensive grasslands have been reported from this assemblage, the result of an incorrect identification. This topic is discussed further below.) At this time, Lake Frome was a freshwa- ter lake with a wide, swampy border (Callan 1977), with rainforest in the hinterland. A ?Miocene assemblage from the Ti Tree Basin, northwest of Alice Springs has abundant Nothofagus, swamp taxa and some grass pollen (discussed further, later) (Kemp 1978; Truswell and Harris 1982). An ?early-mid Miocene assemblage at Lake Hydra, near Lake Eyre, has minimal rainforest gymnosperms (araucarians and podocarps), the sclerophyllous Casuarinaceae and Eucalyptus, and abundant swamp taxa (Martin unpubl.). Thus there was more sclerophyllous vegetation and limited rainforest in Central Australia in the ?early-mid Miocene, when compared with southeastern Australia, and grasses were rare. Evidence about the vegetation of the late Tertiary is very limited, but a few records exist. An ?early Pliocene assemblage at Lake Frome has abundant Casuarinaceae, rare Eucalyptus and a diversity of swamp taxa. There are no rainforest taxa present (Martin 1990b). The vegetation was thus entirely sclerophyllous. A ?late Pliocene-Pleistocene assemblage in the Lake Eyre Basin has low frequencies of Casuarinaceae and Myrtaceae, the only possible trees, and abundant Asteraceae (daisies), the chenopod type (saltbush- bluebush), Poaceae (grasses) and Cyperaceae (reeds). The vegetation was open shrub- lands (Martin unpubl.), not unlike the present day arid zone vegetation (Sluiter and Kershaw 1982). The record in Central Australia shows the following progression when compared with southeastern Australia: 1. Late Paleocene-late Eocene: Varied rainforests were predominant and there were patch- es of sclerophyll vegetation. The climate was somewhat drier than that in the southeast. . Late Oligocene-mid Miocene: Sclerophyllous vegetation was well represented but rainforest was still present in the landscape, though much less than in southeast Australia where it was predominant. Swamp taxa were common. 3. ?Early Pliocene: Sclerophyllous vegetation predominated with little or no evidence of rainforest. Rainforest persisted in the more favourable habitats in the southeast. 4, ?Late Pliocene-Pleistocene: The vegetation had become open shrubland, with a signif- icant herbaceous content and more like the arid vegetation of today. Forests were declining in the late Pliocene of the southeast. tO Proc. LINN. SOc. N.S.W., 119. 1998 H.A. MARTIN 125 el a —) q Uy) ~ a 250 My \ Ls i] 2 250 7D Ne pe Arid tussock grasslands S Arid hummock grasslands /) ro) fe) (e) Coastal grasslands Sub-humid tropical grasslands Isohyets (mm) Sub-humid temperate grasslands BOY oe ee OE OO Sub-humid sub-alpine F}— f°) 20,40 60 80 km grasslands —=— Figure 4. The distribution of grasslands in Australia and the annual precipitation. From Moore (1973) and Leeper (1970), respectively. THE DEVELOPMENT OF GRASSLANDS Today, grasslands predominate in arid and semi-arid regions of Australia (Moore 1973 and Fig. 4). The development of grasslands is of special interest, for without grass- lands, there would be few, if any, grazing animals. The vertebrate palaeontological record shows that there were no grass-eating animals until the mid Miocene, when they were rare, and they did not become significant until the early Pliocene (Archer et al. 1989). Grass pollen first appears in the fossil record in the Palaeocene (Muller 1981) but it is rare. Grasses and other small ground covering plants cannot flourish under a forest canopy because the forest floor is too shaded for good growth. An abundance of grasses may indicate grasslands, but there are many species of swamp grasses (Sainty and Jacobs 1981). Grasses may be abundant in salt marshes as well. Most grasses are wind pollinated and are thus high pollen producers, contributing high frequencies to the pollen assemblage. Some grasses however, are cleistogamous, 1.e. pollination takes place in closed flowers, hence pollen is not liberated in the air. The recognition of grasslands is based on the pollen assemblage as a whole, and not simply the frequency of Poaceae. If open vegetation has grasses of the type which do not con- tribute large quantities of pollen to the assemblage, then other herbaceous taxa, e.g. Asteraceae, are indicative of open vegetation. Proc. LINN. SOc. N.S.W., 119. 1998 126 TERTIARY CLIMATE IN AUSTRALIA ° Ww Ww c , 3 © SSS SS SS SS SS SS SS = ” == =) o@ & -_— —_—— —_! —— ee oo oo ——— = {a0} esi = wW 2 ao & = LW O ee} C) a oO w ie i —S (¢p) bh oT a0} ® > c a @ ea ee ne) = ° > LL za Lu O < is < O Siry ¢@ oo = © = ag = za am LW <9 =) (G} Lu za LWW O © paul oO wo YP (40) fab) > (= Oo = 5 A wre D O i= S oO (ab) Qa 7p) Ww = ee Figure 6. The frequency distribution of phytoliths originating mainly from grasses (see text) found in a deep sea site on the Lord Howe Rise. From Locker and Martini (1986). Proc. LINN. SOC. N.S.W., 119. 1998 H.A. MARTIN 129 Middle Miocene Kaolinite —.p Westerlies ——®» Tropical cyclones = Surface currents Kaolinite 20° 1 { \ \ \ 588 | 590 1 1 oo 1 @591*, 1 \ — 7a Soret \ \ l x G. 40° ~ . ‘, Illite ay , \ / C7 Figure 7. The expansion of aridification southwards in the Miocene, as interpreted from the S/I ratios of clay mineral assemblages on the Lord Howe Rise. From Stein and Robert (1986). swamps etc. for pollen preservation, the few sites in arid/semi-arid regions that have been studied probably only record the wettest periods of the time (see the sea level curve in Fig. 2) and the wettest habitats. There is however, substantial evidence about the development of aridity from other studies, and the most pertinent of these are reviewed. Windblown dust settling in the ocean is a source of evidence about deflation from the source area. Several deep sea sites on the Lord Howe Rise (Fig. 7) record Tertiary dust particles transported by westerly winds from Australia (Stein and Robert 1986; McTainsh 1989). These sites may collect terrigenous particles carried by ocean currents, or by winds from New Zealand. Volcanism is another source of dust. Today, the aeolian sediment influx is dominated by westerly winds bringing dust from the Australian deserts and semi-deserts, and all the evidence suggests that this was the case throughout the Neogene (Stein and Robert 1986). Proc. LINN. Soc. N.S.W., 119. 1998 130 TERTIARY CLIMATE IN AUSTRALIA Site 588 Site 590 Site 591 S/I ratio S/\ ratio S/I ratio CLIMATIC INTERPRETATION 0 8 16 0) 8 16 Dominantly arid climate Major expansion of deserts and semi-deserts * Aberration A A A A A A A A Transition from dominantly humid to a more semi-arid climate Million Years Warm climate, alternating humid and arid periods Ain Low S/I ratio G High S/I ratio A indicates aridity Figure 8. The smectite/illite (S/I) ratios of clay mineral assemblages in deep sea sites on the Lord Howe Rise. Black arrows mark the change from predominantly high to extremely low S/I ratios and indicate the beginning of aridification. *This aberration in Site 591 is thought to have originated from volcanic activity in New Zealand. From Stein and Robert (1986). For location of the sites, see Fig. 7. Once the climate becomes drier and the vegetation is disturbed, wind erosion increases. The dust eroded from sand and stone deserts is less than that from semi-arid regions, but it is greater than that from humid regions. The size of the particles gives an indication of wind speeds. Different types of clay minerals may be used as palaeoclimat- ic indicators also. Illite is formed from predominantly physical weathering. Kaolinite is formed from chemical weathering in tropical areas. Kaolin rich soils in the arid regions of Australia were formed in more humid climates. Smectite is formed in humid to semi- arid climates. Today the snowfields in southeastern Australia collect mainly illite and kaolinite from the arid regions to the west. Dust of volcanic origin is identifiable (Stein and Robert 1986). Figure 8 presents the smectite/illite (S/I) ratios for three deep sea sites on the Lord Howe Rise (from Stein and Robert 1986). Low ratios indicate aridity, as shown by ‘A’ on Fig. 6. In the latest Oligocene, the climate of the source areas was warm with alternating periods of humidity and semi-aridity. In the mid Miocene, the decrease in the S/T ratios at Site 588 suggest increased aridification in the northern to central parts of the continent. Southern Australia may still have been dominated by alternating humid and semi-arid Proc. LINN. Soc. N.S.W., 119. 1998 H.A. MARTIN 131 climatic conditions. The decrease in the S/I ratios in the late Miocene at Site 590 is inter- preted as an expansion of aridification southwards as shown in Fig. 7 (Stein and Robert 1986). This late Miocene southwards extension of aridification coincides with the late Miocene low sea level and the palaeobotanical evidence for a drier climate. The evidence from clay mineralogy suggests that aridification started in the north of the continent and expanded southwards (Fig. 7). Lake Frome is now a dry salt lake, but in the early Miocene it was a large freshwa- ter lake with a wide swampy margin. A study of the clay mineralogy shows that in the early Miocene, the climate was sub tropical. There was a trend towards aridity from the mid Miocene. Marked climatic fluctuations, which may have been seasonal, were super- imposed on this overall trend. The climate approached the present Mediterranean type in the medial Pleistocene (Callan and Tedford 1976; Callan 1977). These interpretations for Lake Frome are in accord with the hypothesis of Stein and Robert (1986) for the south- ern part of the continent. The sediments of Lake George in southeastern Australia (Bowler 1982) have been palaeomagnetically dated to the last 2.5 million years. Initially, Lake George shows con- tinuous deposition of laminated clays representing widespread humid and equitable con- ditions over its region. About 5—6 million years ago, there was a drastic change: the lake dried out and the sediments became deeply weathered. It is thought that this change occurred at the time of the late Miocene low sea level. The climate was seasonally arid to allow the lake to dry out, but with summer rainfall sufficient to promote the deep weath- ering. High lake levels returned briefly at 2.5 million years ago and from this time on, the lake levels oscillated between lake-full and lake-dry conditions. This pattern continues to the present, with amplifications in the last 0.7 million years. These oscillations heralded the onset of the cooler temperatures of the Pleistocene and the glacial/interglacial cycles (Bowler 1976, 1982). From these studies of Lake George and elsewhere, Bowler (1982) advocates that aridity developed from the south. It appears that Bowler’s hypothesis contradicts that of Stein and Robert (1986) who postulate that aridification developed first in the north and expanded southwards (see Fig. 7). The two hypotheses, however, apply to different times. The hypothesis proposed by Stein and Robert is based on evidence for the mid- late Miocene, from 14-15 to 5—6 million years ago (Fig. 8), whereas Bowler’s hypothe- sis relies on evidence for the period >6—2.5 million years to the present. The early stages of aridification may have been strongly seasonal (Callan 1977) and a well-marked dry season would be sufficient to generate the dust storms necessary to transport dust out to sea, even if there was a wet season as well. The S/I ratios (Fig. 8) suggest that aridity intensified about 3 million years ago, and this part of Stein and Robert’s evidence is in accord with Bowler’s evidence from Lake George. Another hypothesis places the beginnings of aridity in the Eocene and uses evi- dence that the change from terrigenous to carbonate sedimentation off the western mar- gin of the continent infers cessation of efficient drainage from the land (Quilty 1982). The lack of terrigenous sediment, however, only infers minimal erosion from a flat, well- vegetated landscape. There is contradictory evidence to refute this hypothesis. Laterite formation requires a warm, wet climate and there is more than one episode of lateritisa- tion. Palaeomagnetic dating suggests that the dominant period of laterite weathering over a large part of Australia was late Oligocene to early Miocene (Idnurm and Senior 1978; Schmidt et al. 1976). Moreover, river systems in the arid region of Western Australia have been inactive only since the mid Miocene (van der Graff et al. 1977). This latter evidence suggests that the trend towards aridity did not start until the mid Miocene on the western margin of the continent. Beard (1977) presents a hypothesis that aridity started in the northwest of the con- tinent in the mid Eocene, some 45 million years ago, on the basis that this part of Australia was the first to reach the dry anticyclone belt by continental drift, i.e., it drifted Proc. LINN. SOc. N.S.W., 119. 1998 132 TERTIARY CLIMATE IN AUSTRALIA into aridity. The evidence presented above does not support this hypothesis. Kennett (1993) considers that continental drift alone is too slow to account for the climatic changes observed . In a study of foraminifera and sedimentation off northwest Australia, Apthorpe (1988) concludes that the region was a warm water to tropical province throughout most of the Cainozoic. The evidence suggests that there were dry periods and perhaps season- ally dry climates at times during the late Palaeocene and late Eocene, but not the begin- ning of the aridity seen today. The undated palaeobotanical evidence from Melville Island (Pole and Bowman 1996) also suggests a seasonal climate. Aridification intensified with the Quaternary period which is dominated by the glacial/interglacial cycles. The glacial periods were colder, windier and more arid (see Kershaw 1981; Kershaw et al. 1991; Dodson 1992, 1994; Hope 1994). Desert dunes were more mobile (Wasson 1989), the arid zone expanded in size (Dodson and Wright 1989) and the dust flux in marine sediments increased (Hesse 1994). About half a mil- lion years ago, there was a further increase in aridity to that resembling today (Bowler 1982). Palaeobotanical records only exist for the latter part of the Quaternary. In south- eastern Australia, the vegetation was predominantly open shrubland/grassland/herbfield during glacial times and forested in the interglacial periods (Dodson 1992; Hope 1994). In northeast Queensland, sclerophyll vegetation was dominant in the glacial cycles and rainforest in the interglacials (Kershaw 1985, 1994). In the last glacial period, the Gulf of Carpentaria was exposed and a savannah-like environment prevailed. The runoff/evaporation rates were about half of the present ratio (Torgersen et al. 1988). In northwest Australia, the last glacial period was distinctly drier and grasslands replaced the eucalypt forest of the previous interglacial (van der Kaars 1990). It is beyond the scope of this study to deal with the Quaternary in detail, but the constantly changing environment, on a scale of thousands of years, must have had a profound influence on the biota. The extensive Tertiary vertebrate faunas of Riversleigh (Archer et al. 1989, 1994) carry a climatic signal but there are few palaeobotanical records anywhere near Riversleigh (Fig. 1) and extrapolation from the few records in the northwest and north- east must be done with great caution. Only the late Quaternary study, covering the last 35,000 years from the Gulf of Carpentaria (Torgersen et al. 1988) is geographically close to Riversleigh. Today, Riversleigh experiences a monsoonal climate and while the mechanics of the present day monsoonal system is well researched, very little is known about its history (Crisp 1996). There is some evidence of seasonal climates in the early Tertiary from both northern (Apthorpe 1988; Pole and Bowman 1996) and Central Australia (Callan and Tedford 1978; Greenwood 1996), but it is not known if it was monsoonal. CONCLUSIONS In southeastern Australia, there are two periods of dramatic change, one in the mid Miocene, when widespread rainforest disappeared and sclerophyllous forests became dominant, and the other in the late Pliocene-Pleistocene, when the forests declined and open vegetation became dominant. The Tertiary precipitation curve shows long periods of relative stability and short periods of marked change. Other parts of the world show marked climatic change about the same time. These times of change may be correlated with global changes in sea lev- els and climate. When all the sites in southeastern Australia are compared, they indicate that it was wetter in Tasmania and on the southern highlands, and drier inland and to the northwest, i.e., during the Tertiary, there was a climatic gradient parallel to that of today. Proc. LINN. SOc. N.S.W., 119. 1998 H.A. MARTIN 133 Northeast Queensland shows relative stability of the rainforest/sclerophyll vegeta- tion from the late Miocene through most of the Pleistocene, when compared with south- eastern Australia. The precipitation was maintained above the limits for rainforest throughout the period, hence northeast Queensland became a refuge for taxa which could not tolerate the drying environment elsewhere. Northwest Australia shows a progression from casuarinaceous forests to shrub- lands/grasslands through the late Miocene-Pleistocene, very different to that of the south- east which still had appreciable rainforest. The Eocene of Central Australia had both rainforest and sclerophyllous vegetation, and the climate was seasonal. Rainforest decreases earlier in Central Australia than in southeastern Australia. When the vegetation and climate of all these different parts of Australia are com- pared, they suggest that a general climatic gradient parallel to that of today, existed throughout the Tertiary. Grasses were rare in the early Tertiary and started increasing in the late Miocene. Grasses increase in the Pliocene of central regions of Australia, and in the late Pliocene—Pleistocene of southeastern Australia. The trend towards aridity is first detected in the mid Miocene. The late Pliocene- Pleistocene witnessed a marked increase in aridification and the present day level of arid- ity was reached about a half million years ago. ACKNOWLEDGEMENTS I am indebted to Prof M. Archer who stimulated this study which was presented at the Riversleigh Symposium, Sydney 1994. 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SEMPLE” AND T.B. KOEN? ‘Department of Natural Resources and Environment, PO Box 275, Benalla, Victoria, 3672 *Department of Land and Water Conservation, PO Box 53, Orange, NSW 2800; and * Department of Land and Water Conservation, PO Box 445, Cowra, NSW 2794 LAWRENCE, J., SEMPLE, W.S. AND KOEN, T.B. (1998). Experimental attempts at encouraging eucalypt regeneration in non-native pastures of northern Victoria and central western NSW. Proceedings of the Linnean Society of New South Wales 119, 137-154. This paper reports the results of three “cartwheel-type” regeneration experiments around woodland eucalypt trees (Eucalyptus melliodora and E. albens). In each experiment treatments such as scalping (removal of topsoil) and applying herbicides were applied to plots radiating from the trunk of a tree and were replicated in each of the four main compass direc- tions. Scalping at Mansfield and herbicide/mowing at Panuara, resulted in significantly higher numbers of seedlings but spatial variability in the amount of regeneration around the trees was also observed with seedlings more common on the southern sides of trees. The conven- tional explanation for this, viz. unequal seed dispersal due to direction of prevailing winds, does not adequately explain the results. It is suggested that habitat factors, such as differences in shading or topsoil moisture, may offer a better explanation — at least in seasons which are less than optimal for regeneration. Regeneration did not occur in every season despite the apparent receptivity of seedbeds. The results suggest that if seedfall is adequate and a recep- tive seedbed is present, then above-average spring and December rainfall, together with at least 80 mm of rain in January, February or March, are necessary for successful spring/sum- mer recruitment. Climatic conditions were suitable for regeneration at Mansfield (Victoria) and Panuara (NSW) in 1992/93. One year old scalps were associated with the highest number of seedlings at Mansfield but newly-prepared scalps at Panuara were associated with the least number of seedlings during the same season. Possible reason for these results are discussed. Manuscript received 20 July 1997, accepted for publication 22 October 1997. KEYWORDS: Eucalyptus albens, Eucalyptus melliodora, recruitment, shade, scalping, rain- fall, glyphosate. INTRODUCTION The case for re-establishing trees and shrubs in the rural environment has been put forward by many authors, including Beckmann and Davidson (1990), Bird et al. (1992) and Dalton (1993), and will not be repeated here. Tree and shrub establishment technolo- gy has improved remarkably in recent years. As a result of research in southern Australia, e.g. by C.V. Malcolm in Western Australia, G. Dalton in South Australia and P.R. Bird in Victoria, reliable direct seeding techniques are now available for rural lands (Dalton 1993). Aerial seeding of eucalypts has also been successfully practised on mined sites in areas with predictable seasonal rainfall (e.g. Foster 1985) and some success with simulated aerial seeding on rural lands was reported by Campbell and Nicol (1996). Natural regeneration has been the basis of forest regeneration following logging for many years (Florence 1996) and seed production and regeneration requirements of most commercial forest eucalypts are reasonably well known (e.g. Boland et al. 1980, Cremer et al. 1990). In rural areas, however, natural regeneration has been limited by grazing, cultivation and competition from pasture species for over a century and many of Proc. LINN. SOC. N.S.W., 119. 1998 138 EUCALYPT REGENERATION IN PASTURES the remaining trees are very old (Middleton 1984, Campbell et al. 1988) . Regeneration is dependent on the chance occurrence of heavy seedfall, high soil moisture and suitable temperatures for germination (Venning 1988) as well as a favourable weed-free seedbed. Curtis (1990) estimated that these conditions occurred only once every 10 to 20 years in the Northern Tablelands of NSW. Attempts at encouraging natural regeneration in rural areas have been disappointing but unplanned occurrences do occur (Venning 1988, MacLennan et al. 1992). In the higher rainfall country of South Australia, natural regen- eration was associated with low feral animal populations, low grazing pressure by domestic animals, the presence of scattered trees, lack of “pasture improvement’, and above-average rainfall for two consecutive years (Venning 1985). Reducing herbage competition, e.g. by cultivation and applying herbicides prior to expected seedfall, and controlling grazing following emergence, have been suggested as ways of enhancing regeneration (Venning 1988, Curtis 1990, Cremer et al. 1990). Venning (1988) also suggested concentrating efforts on situations where regeneration was more likely such as around species known to be good regenerators, e.g. Eucalyptus camaldulensis, where exotic pastures are absent, and on the leeward side of tree clumps. Until reliable techniques for promoting natural regeneration in woodlands have been developed, infrequent occurrences of regeneration can be protected, e.g. as was reported by Semple (1987). In some areas regeneration may already be present in the form of sup- pressed lignotuberous seedlings, which can be encouraged by altering the grazing regime (Curtis 1990, Cluff and Semple 1994). Unlike commercial forest species, recruitment processes in woodland eucalypts are poorly understood (Venning 1988). Some progress on flowering, seedfall, seed predation and/or emergence processes has been made in recent years for Eucalyptus salmonophloia F. Muell. (e.g. Yates et al. 1996), eucalypts of the Northern Tablelands of NSW (Curtis 1989, 1990), some tropical woodland eucalypts (Setterfield and Williams 1996) and some eucalypts of the Central West of NSW (Semple and Koen 1997). In the last-men- tioned study of eucalypt emergence and survival on differently prepared seedbeds, it was found that no particular seedbed was consistently associated with high numbers of emer- gents. The authors suggested that the main effect of seedbed preparation was in control- ling exotic herbage competition during the eucalypt seedling’s first year. The aim of the experiments described below was to evaluate the role of seedbed type on eucalypt emergence and survival around actual trees, rather than in small plots divorced from the woodland situation. As with Semple and Koen’s (1997) study, seedbeds were prepared in understories dominated by exotic species, which are common in the winter rainfall zone of south-eastern Australia. Unlike the former study, however, these experiments were designed to detect effects of the presence of the parent tree on emergence/survival, where seed supply was not controlled, although it was monitored in one of the experiments. The species selected for the experiments, yellow box (Eucalyptus melliodora Cunn. ex Schauer) and white box (E. albens Benth.), are com- mon woodland species of the inland slopes of south-eastern Australia. METHODS A common feature of the three trials was the imposition of a range of treatments in plots that radiated out in a “cartwheel” fashion from the trunk of a relatively isolated tree (e.g. Figure 1). Each seedbed treatment was repeated in each of four quadrants as defined by the main points of the compass. In each study it was believed that fruits on the trees selected would shed seed during the period of the trial. All trial areas were fenced to exclude domestic stock. Climatic and other data for the three trial sites are presented in Table | and Figure 2. As the Mansfield trial was carried out independently of the other two, there were some differences in the methods used. Proc. LINN. SOC. N.S.W., 119. 1998 J. LAWRENCE, W.S. SEMPLE AND T.B. KOEN 139 a Sc scalp Dec 90 + glyphosate Nov 91 E.camaldglensis ) Sl slash Dec 90 + Nov 91 q eo) Cu cultivated Dec 90 + glyphosate Nov 91 Ws Nil no treatment Zz Oueen See en Olin See Zorn ke o) E — SCALE Me oe Figure 1. “Cartwheel” layout of treatments at the Mansfield site. Mansfield (Dec. 1990 to May 1994) This experiment was located 9 km west of Mansfield in the foothills country of north-eastern Victoria. The area around an isolated Eucalyptus melliodora tree was divided into four quadrants: NW—NE, NE-SE, SE—SW and SW—NW. Each quadrant was subdivided into four plots extending 30.5 m from the centre of the trunk of the tree (Figure 1) and the following treatments randomly allocated to the 5.5—-30.5 m zone in early December 1990: scalping — removal of the top 3 cm of soil; slashing and removal of slashed material; cultivation to a depth of 10 cm with a rotary hoe; and control — no treatment. As no seedlings were evident by early November 1991, a further set of treatments was imposed such that overall treatments were: scalping Dec. 1990 and glyphosate at 2 L/ha in Nov. 1991; slashing Dec. 1990 and Noy. 1991 and removal of slashed material; cultivation Dec. 1990 and glyphosate at 2 L/ha in Nov. 1991; and control — no further treatment. Proc. LINN. SOc. N.S.W., 119. 1998 140 EUCALYPT REGENERATION IN PASTURES TABLE | Brief details of trial sites Mansfield Panuara Molong Elevation (m) 35 ew Ny 580 FAS Has 540 Mean annual rainfall(mm) 717 (Mansfield PO) 748 (“Weemalla’ ) 705 (Molong) Land use (grazing by:) sheep & cattle sheep sheep Pasture perennial ryegrass, introduced annuals phalaris, sub. clover and sub. clover and other other annuals introduced annuals Slope (%) 4-5 8-13 4-14 Aspect north-east west Tree ‘A’ (east) Tree ‘B’ (west) Panuara (Oct. 1992 to Feb. 1995) This experiment was located 30 km south-west of Orange in the Central Tablelands of NSW. Relatively isolated E. melliodora trees were again the subject of investigation. Selection of trees was constrained by the small size of the ungrazed paddock. Hence, truly isolated trees were difficult to obtain. One tree and half of the area around two other trees were selected and treatments imposed on radiating plots as before, except that quadrants were bounded by the cardinal points of the compass, i.e. N-E, E-S, S—W and W-N. Plots extended for 9.5 to 10 m beyond the mean canopy radius of each tree. The following treatments were randomly allocated to one of five plots in each quadrant in late 1992: scalping — removal of the top 10 cm of soil (Nov.); glyphosate at 1 L/ha (early Oct.)/slashing (mid Oct.); glyphosate at 1 L/ha (early Oct.)/slashing (mid Oct.)/fluazifop-p, a grass-selective herbicide, at 4.3 L/ha (Jan. 1993); glyphosate at 1 L/ha (early Oct.)/slashing (mid Oct.)/fluazifop-p at 4.7 L/ha + simazine, a residual herbi- cide, at 3 L/ha (Feb. 1993); and control — glyphosate at | L/ha (early Oct.). A preliminary examination of seedling numbers suggested that seedlings were not evenly distributed around trees — an effect which may have been due to summer shad- ing. Shading on the treated area around each tree was mapped hourly on a cloudless day at Panuara in midsummer (14 Jan. 96) and at Mansfield in late summer (15 Feb. 96). Due to differences in mapping scales, timing of observations, diameters of treated areas, aspect, slope, as well as the confounding influences of surrounding trees, it was not pos- sible to directly compare results at the two sites, nor to analyse them in the same way. Molong (Aug. 1993 to Feb. 1995) The experiment area was 9 km west of Molong near the western margin of the Central Tablelands of NSW. The circular area around two isolated Eucalyptus albens trees was subdivided into four quadrants: NW—NE, NE-SE, SE-SW and SW-NW. Each quadrant was subdivided into three plots extending 5 m beyond the mean radius (6.6 m and 8.5 m) of the canopy of each tree. Glyphosate was applied at about 2 L/ha to all plots in mid Aug. 1993 and to any missed areas in early Sep. 1993. One of the objectives of this trial was to evaluate the effects of staggered monthly follow-up applications of glyphosate until germination seemed imminent, i.e. when seed- fall and rainfall coincided. Treatments were randomly allocated within each quadrant as follows: follow-up glyphosate one month (late Sept. 1993) and four months (mid Jan. Proc. LINN. SOc. N.S.W., 119. 1998 J. LAWRENCE, W.S. SEMPLE AND T.B. KOEN 141 Mansfield (Vic) 400 300 200 100 400 300 200 Rainfall (mm) 100 400 __ Molong (NSW) 300 200 100 Figure 2. Seasonal rainfall (mm) at the three sites during the periods of the trials. Thickened lines indicate long term means at Mansfield (1901-93), ““Weemalla”, Panuara (1971-92) and Molong (1884-1993). Thin arrows indicate the commencement of treatments (and retreatments at Mansfield) and thick arrows indicate the first occurrence of seedlings at each site. 1994) later; follow-up glyphosate two months later (early Nov. 1993); and follow-up glyphosate at three months later (mid Dec. 1993). The second objective of this experiment was to monitor some factors thought to be responsible for recruitment. Circular 0.25 m? seed collectors (Figure 3) were installed at the edge of the canopy around both trees and monitored at approximately weekly inter- vals. Two collectors were located in north and south positions at one tree (“A’) and four at north, south, east and west positions at the other (“B’). Maximum/minimum temperatures, Proc. LINN. SOc. N.S.W., 119. 1998 142 EUCALYPT REGENERATION IN PASTURES Figure 3. A simple cloth mesh (2125 LaCoste fabric manufactured by Petlee P/L) seed collector at Molong. A stone was placed in the bottom of the collector to prevent its being disturbed by wind. as recorded by a shielded thermometer in the canopy 4.5 m above ground level, and also at the nearby homestead, were recorded at approximately weekly intervals. Topsoil (O—5 cm) moisture (gravimetric) was measured in all plots in Oct. 1993 and Feb. 1994. Statistical Analyses Data from these trials were analysed using analysis of variance techniques, trans- forming the data by taking natural logarithms where necessary. Data based on treatments applied about a single tree used the compass based quadrants as blocks, necessitating a residual error term based on treatment by quadrants interaction. For sparse data such as from the Panuara trial, a generalised linear model was used under the assumption of a Poisson error distribution and log link function. Treatment means were examined for sig- nificant differences (p = 0.05) using the protected least significant difference procedure. RESULTS Seedlings and Survival Seedlings of E. melliodora (and some of E. camaldulensis) were first observed at Mansfield in December 1992, 24 months after initial treatment and 13 months after retreat- ment. At Panuara, E. melliodora seedlings were first observed in January 1993 about 3 months after initial treatment. Only one E. albens seedling was observed at Molong, in February 1994, 5 months after initial treatment; this seedling did not survive. In contrast, most of the summer 1992/93 seedlings at Mansfield and Panuara were still present at the end of the second summer (and third summer at Panuara). Further recruitment occurred at Mansfield in summer 1993/94 but it was less marked at Panuara (Table 2). Proc. LINN. SOc. N.S.W., 119. 1998 J. LAWRENCE, W.S. SEMPLE AND T.B. KOEN 143 TABLE 2 Total numbers of seedlings recorded around each tree (across all treatments) at the three trial sites Site/trees/ NDB Number of seedlings times of treatment area (m’) i 4 a ae, = aa ied treated autumn autumn autumn autumn late summer 199] 1992 1993 1994 1995 Mansfield (Dec. 1990 2830 0 0 205 299 = & Nov. 1991) Panuara A 1180 83 104+ 1107 Panuara B* 515 10 14+ 15 Panuara C* (Oct. 1992 450 1 1 it to Feb. 1993) Molong A 285 0 0 Molong B (Aug. 1993 345 | 0 to Jan. 1994) * Only half of the area around each tree was treated. + Seedlings present in 1994 were of a similar size. It is likely that many of the additional seedlings were missed in the 1993 counts. + Change from 1994 was due to 9 new seedlings (of which 5 were on scalped plots) and 3 deaths. Seedlings not counted Distribution of Seedlings For seedling counts at the Mansfield site, plots were subdivided at radial intervals of 5 m. This permitted an assessment of the effect of distance from the tree on seedling numbers. When averaged across treatments, seedling density was significantly (p = 0.05) higher at and just beyond the edge of the canopy (Figure 4). Where all of the area around a tree was treated, directional effects were also appar- ent (Figure 5) [as treatments were only imposed beyond the edge of the canopy at Panuara, seedling numbers beneath the canopy at Mansfield have, for comparison pur- poses, been omitted from this figure]. At Mansfield, the treatment with the highest num- ber of seedlings (scalping) accounted for 72 % of all seedlings and was largely responsi- ble for the distribution shown in Figure 5. As described later, differences between treat- ments were less marked at Panuara. However, when the proportions of seedlings are pre- sented in equivalent quadrants to those at Mansfield (though not all treatments are now equally represented in each quadrant), a similar pattern for the north and south quadrants emerged — but proportions in the east and west were reversed. Due to the lack of repli- cation at either site, the statistical significance of these patterns cannot be quantified, but a higher proportion of seedlings in the south is strongly suggested. Shading of Plots At Panuara, the area of each plot which was continuously shaded between the hourly observations was estimated using grid squares. These were summed and divided by plot area to give ‘full plot shade hours’ for each plot (Figure 6A). A simpler procedure was used for the Mansfield data. Each of the 13 shade maps was replotted on transparent films, which were then overlaid to provide a map of relative total shading around the tree (Figure 6B). It should be noted that, unlike the Panuara data, this diagram includes a considerable area beneath the canopy. Proc. LINN. Soc. N.S.W., 119. 1998 144 EUCALYPT REGENERATION IN PASTURES Mean canopy radius = 11.6m Seedling nos. (m*) SS 10.5 15.5 20.5 25.5 30.5 Distance from tree centre (m) Figure 4. Seedling density around the E. melliodora tree at Mansfield in autumn 1994 — direction and treat- ment effects ignored [Tree silhouette is diagrammatic only]. North 7% S canopy of t 10% 40% 28% My) 10% 7 N A 49% ‘ 43% aan 10m Figure 5. Numbers and proportions of seedlings outside the canopy in each of the four treatment quadrants at Mansfield (M) and Panuara (P) in autumn 1994. For comparison purposes, seedlings under the canopy at Mansfield have been omitted. Proc. LINN. SOc. N.S.W., 119. 1998 J. LAWRENCE, W.S. SEMPLE AND T.B. KOEN 145 Figure 6. Two methods of comparing the rela- tive amounts of total daily shading on treated areas at Panuara and Mansfield: A. Cumulative hourly continuous shade in each treatment plot between mean canopy edge and 10 m away at Panuara, 0730 to 1930 DST, 14 Jan. 1996. See text for explanation of units. B. Overlay of shaded areas recorded each hour in the treated areas between 5.5 and 30.5 m from the centre of the tree at Mansfield, 0700 to 1900 DST, 15 Feb. 1996. b Bead North Full plot shade hours Proc. LINN. Soc. N.S.W., 119. 1998 146 EUCALYPT REGENERATION IN PASTURES At Panuara in midsummer, plots in the north-east and west received the longest period of shading. Comparison of data in Figures 5 and 6A, however, suggested that midsummer shading and numbers of seedlings were unrelated (Rank correlation = 0.33). The qualitative Mansfield data in Figure 6B also showed considerable shading in the east and west but with increased shading (compared to the data collected a month earlier in Fig. 6A) in the south — consistent with the apparent northerly movement of the sun following the summer solstice. Seedbed Treatments Sufficient numbers of seedlings were present only at Mansfield and Panuara (one full tree and one half-tree) for an adequate analysis of treatment effects. Results are pre- sented in Table 3. The scalp treatment was associated with the most seedlings at Mansfield but with the least seedlings at Panuara. Regeneration was almost immediate in all treatments except scalped plots at Panuara, whereas two years elapsed before it was evident on scalped (and other plots) at Mansfield. Though subsequent recruitment was low at Panuara, it did not occur on scalped plots until some two years after the initial treatment (Table 3). Regeneration occurred in the other three treatments at Mansfield but seedling num- bers were lower than in the scalped plots and not significantly different from each other (Table 3). At Panuara, seedling numbers (Table 3) were significantly higher on the glyphosate/slash treatment and were significantly lower on the scalped treatment, com- pared to the other three treatments. Differences in seedling numbers between the other three treatments were not significant. TABLE 3 Seedling numbers in each of the seedbed treatments at Mansfield and Panuara Site Treatment Back transformed mean numbers of seedlings per plot* Apr. 93 May 94 Mansfield scalp (Dec. 90) + glyphosate (Nov. 91) 25.3at 39.9a (1 tree) slash (Dec. 90 + Nov. 91) 2.1b 5.0b cultivation (Dec. 90) + glyphosate (Nov. 91) 4.2b 5.6b control — no treatment 1.8b 1.5b Mar. 93 Mar. 94 Feb. 95 Panuara scalp (Nov. 92) Oa Oa 0.9a (1% trees) glyphosate/slash (Oct. 92) 6.7b 8.0b 8.0b glyphosate/slash (Oct. 92) + fluazifop (Jan. 93) 3). SIC 4.0c 4.0c glyphosate/slash (Oct. 92) + fluazifop/simazine (Feb. 93) SC 4.5c 4.5c control — glyphosate (Oct. 92) 1.7¢ 3.0c 3.3¢ * seedling numbers are not comparable between sites due to different plot sizes and methods of analysis Means followed by different letters within a site/time subset are significantly different (p<0.05) Proc. LINN. SOc. N.S.W., 119. 1998 J. LAWRENCE, W.S. SEMPLE AND T.B. KOEN 147 Tree A Oct 93 — - Tree A Feb 94 ---- Tree B Oct 93 — Tree B Feb 94 eumme Average Moisture % Oo ol Oo oh, oO oO 150-200, 250 (3005 350 East West North South Figure 7. Mean gravimetric 0-5 cm soil moistures, corrected for treatment effects, in radial plots around two E. albens trees on two occasions at Molong. Topsoil Moisture Moisture (0-5 cm) was measured around the two trees at the Molong site on two occasions — early in the trial (mid Oct. 1993) and shortly after the first summer germi- nations of herbage were observed in February 1994. No treatment was consistently asso- ciated with high or low topsoil moisture levels on either occasion. However, the amount of soil moisture was significantly associated with position around the trees, tending to be higher in the eastern and to a lesser extent, the southern quadrants. Mean soil moistures, with any treatment effects removed, are shown in Figure 7. Seedfall Seedfall was measured from early October 1993 to early April 1994 at the Molong site. Only five seeds were collected in the two 0.25m° collectors at tree ‘A’ — represent- ing a mean ‘seed rain’ of 0.06 seeds m~ day! over the 185 day period. At tree “B’, with four collectors, a total of 147 seeds was collected over 198 days. Of a sample of 129 of them, 101 (78.3 %) were germinable. Peak seedfall, up to 3.7 seeds m~ day‘, occurred in January/February 1994 (Figure 8). Between early October 1993 and germination in mid February 1994, total seedfall was 116 (c.90 viable) seeds m~. Seedfall and Temperature Temperature data for the lower canopies of the trees at Molong were incomplete due to occasional storm damage to thermometers. However, maxima/minima recorded at Proc. LINN. Soc. N.S.W., 119. 1998 148 EUCALYPT REGENERATION IN PASTURES 6 40 5 . > = Maximum a Oo 30 44 S = g Q =) _— § 3g ® 20 8 ‘= — o By . B 10 2 1 0) Oct'93 Nov'93 Dec'93 Jan'94 Feb'94 Mar'94 Apr'94 Figure &. Rates of seedfall at tree ‘B’ (Molong) and maximum temperature over the same period. the nearby homestead were highly correlated with those recorded in the canopies, partic- ularly at Tree ‘B’ where r = 0.90 for maxima and r = 0.93 for minima. Seedfall at Tree ‘B’ (where sufficient seedfall data were available) was correlated with maxima recorded at the homestead (r = 0.66) but not with minima (r = 0.38). Seedfall generally occurred when maxima >25°C were recorded (Figure 8). DISCUSSION As regeneration depends on seed availability, dispersal to safe sites and favourable climatic conditions for germination and establishment, it is difficult to isolate the role of any one factor even in controlled experiments. However, as observations in these experi- ments were made over a number of years and at different sites, some interpretation is possible. Seasonal Effects Curtis (1989) concluded that for successful late spring/early summer recruitment on the Northern Tablelands of NSW, above-average spring to mid-summer rainfall was necessary. Combining data from Curtis (1989) and Semple and Koen (1997) with those from Mansfield, Panuara and Molong (Table 4) indicated that most cases of successful recruitment were associated with above-average spring and December rainfall. If these conditions were satisfied, survival of at least some seedlings was assured provided one month in January, February or March received about 80 mm or more of rainfall. High pre-germination spring rainfall probably provides sufficient subsoil moisture for survival over summer (Whalley and Curtis 1991). However at Manildra, where simu- lated seedfall was carried out, the below-average spring 1991 rainfall was supplemented by well above (432% of) average February 1992 rainfall and seedlings established. Germination occurred at “Terrible Vale” and “Birrahlee” on the Northern Tablelands, but the below-average spring rainfall was not supplemented by sufficient rainfall in January to March and all seedlings died within four months (Curtis 1989). Proc. LINN. SOC. N.S.W., 119. 1998 149 J. LAWRENCE, W.S. SEMPLE AND T.B. 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SIRA PIQLUQL.,, Sa (6b-) 81 (9¢-) Ip (t-) L6 (Ob+) 8ZI (Ob+) 8bZ $8 “AON : SIH Aqny,, 98/S86I (JOINS sani = z ma: : Be ‘uve = ae : “AON F ase canieaiiatord ee Ss puodaq sunsisiod ———- a useq aaevy Avut Kay) ‘o'1) UONRIOUASAI (WIU) [[eyUTeYy ysnoy}) paarasqo poyst[qrisy ISI} SSUI[P9esS J1eq ous uosvas ‘sasayjuared Ul UMOYS Iv [[eJUIeI URDU WoJy (WUT) sainyedag (76/1661 Ul playsueyy sdeysed 3daoxa) a[quyIeae sem poes pure J[QLINOAL] SLM PdqpdeS dU} ‘SOUS [Je FV ‘SOUS SNOLIVA Je ‘[[BJUTeI YIM JayI980} ‘ouruNs AjIea-Surds aye] Ul JUSWINIOAI ydATVONA JO aIn[ IV} puke sssdon¢ p ATIVE Proc. LINN. SOc. N.S.W., 119. 1998 150 EUCALYPT REGENERATION IN PASTURES As most eucalypts can germinate over a wide range of temperatures (Venning 1988), it was unlikely that temperature was limiting in spring-summer in the woodland environment. But as high temperature and associated soil drying are responsible for heavy losses of unprotected new seedlings in forests (Florence 1996), any factors moder- ating high temperatures would be expected to enhance survival of late spring and sum- mer emergents. Effect of Seedbed The 1992/93 season was clearly advantageous for regeneration of E. melliodora. Even ungrazed controls, viz. glyphosate in mid spring at Panuara and untreated at Mansfield, produced some seedlings. Scalping (+ glyphosate 11 months later) was the most effective seedbed at Mansfield, whereas at Panuara, glyphosate followed by slash- ing was associated with the highest number of seedlings after one year. Surprisingly, scalping was the least effective treatment at Panuara. Possible reasons for this apparently anomalous result were: (a) at the time of emergence, the scalp treat- ment at Mansfield was two years old and herbicide had been applied to remove limited herbage growth. In contrast, the scalps at Panuara were newly- prepared when emergence occurred in the other treatments; (b) slope was higher at Panuara than at Mansfield (Table 1) and a greater depth of soil was removed at Panuara than at Mansfield. The reason for regeneration failure at the Mansfield site in the first (1991/92) sea- son was uncertain, though it was probably due to inadequate rainfall. At Panuara, season- al conditions were favourable in 1992/93, yet seedlings did not appear on scalped plots (which remained relatively bare) until some time between March 1994 and February 1995 (Table 3), despite poor seasonal conditions during this time. These results suggest that scalped areas require a ‘settling in’ period — perhaps due to enhancement of roughness by litter accumulation or by limited herbage growth. This would probably be more important on land of higher slope. The necessity of some ‘vegetative roughness’ for successful aerial seeding of exotic pasture species on infertile sites, and for simulated aerial seeding of eucalypts, has been observed by Campbell (1996) and I. Higgins (DNRE Victoria, pers. comm.) respectively. Vegetative mulches also have other benefits such as reduced evaporation and increased rainfall infiltration (Dalton 1993). Further support for this suggestion comes from Semple and Koen’s (1997) small plot trials where ripping of scalped plots was found to be necessary for retention and ger- mination of surface-sown seed. Although scalping was not associated with the highest number of seedlings initially (compared to other seedbed treatments), it commonly was after one year and was attributed to enhanced survival of initial emergents and the possi- bility of further germinations on a weed-free seedbed. Excess standing vegetative residue, however, reduced regeneration at Panuara, where significantly (p = 0.05) more established seedlings were associated with the glyphosate/slash treatment than for glyphosate alone. Post-emergence applications of a grass-selective herbicide (fluazifop-p), either alone or in combination with a residual herbicide (simazine), to glyphosate/slashed plots (which had low levels of live herbage at the time) did not enhance survival, but, com- pared to the glyphosate/slash treatment, significantly reduced it. Although these herbi- cides were recommended by Bird et al. (1994) for young trees, they were referring to mature seedlings rather than those at the 2 to 4-leaf stage. The role of pre-germination applications of glyphosate in assisting regeneration, particularly at Panuara, but also noted by others such as Curtis (1989), needs some expla- nation as it is not always successful. Applying herbicide around trees commonly fails to promote regeneration because of lack of seed and/or an unfavourable season, such as occurred at the Molong site in 1993/94. Even when these conditions are satisfied and Proc. LINN. SOC. N.S.W., 119. 1998 J. LAWRENCE, W.S. SEMPLE AND T.B. KOEN 151 germination occurs, survival may be low or negligible as was the case in many of the seedbed trials (direct drilled and surface-sown) reported by Semple and Koen (1997). Survival after one year in these trials was generally higher on the scalped treatments than on any of the others, including herbicide. The poor performance of the non-scalp treat- ments was attributed to the rapid recolonisation of plots by weeds. It was suggested that if treatments had been delayed until later in the season, better weed control may have been achieved. The success of glyphosate at Panuara can probably be attributed to the late application (October — due to earlier wet weather) and the reduced frequency of summer weeds at the site. Seedfall Only one of the two E. albens trees at the Molong site consistently produced enough seed for adequate analysis. Peak seedfall occurred in January 1994 when up to 3.7 seeds m~ day ' were recorded. This declined steadily until April when observations ceased. This pattern of seedfall was consistent with observations of other eucalypt species on the Northern Tablelands of NSW by Curtis (1989), who also noted that regen- eration was commonly associated with the ‘rising limb’ of the seedfall graph, rather than at its peak. The reasonable correlation between the rate of seedfall and maximum weekly tem- perature, was not unexpected. However, as dehydration is largely responsible for seed release in eucalypt fruits (Christensen 1971, Boland et al. 1980), low humidity, which would be expected to be associated with higher temperatures in non-coastal areas, was probably the operative factor. Although the rate of seedfall was not high, it was comparable to that associated with successful recruitment in the Northern Tablelands. Here, Curtis (1989) reported that 100 to 200 seeds m~ in the six months prior to germination appeared to be necessary. [It is perhaps not coincidental that a similar number of seeds is also considered desirable for aerial seeding of phalaris (M.H. Campbell, NSW Agriculture, pers. comm.)]. About 90 viable seeds m~ were recorded in the four months prior to germination at Molong. However, only one seedling was recorded (in the SSE) following above-average rainfall in February 1994 but it only survived a few months of below-average rainfall. Distribution of Seedlings According to Cremer et al. (1990), the direction/speed of dry season winds is the main factor affecting seed dispersal of forest eucalypts. Most seed falls within a distance of 1% x height of parent tree, though isolated trees can have a higher proportion of seed beyond this limit than do trees in a forest. In the studies reported here, the treated area did not extend as far as 1% x height of parent. At Mansfield seedlings were recorded up to 30 m from the 33 m high parent which was consistent with Curtis’ (1989) observation of seedlings up to 13 m from an 18 m high woodland eucalypt on the Northern Tablelands. Also consistent with Curtis’ observations was that the highest density of seedlings at Mansfield occurred near the edge of the canopy. As most of the seed falls in this region (Cremer et al. 1990), these observations suggest that the quantity of seed is one of the major factors affecting recruitment. Though only a very small proportion of viable seed (e.g. 0.16%, Curtis 1989) results in established seedlings, studies by O’ Dowd and Gill (1984) indicate that large quantities are needed so that some seed remains after seed-harvesting ants are satiated. If this is the case, then emergence would be almost wholly a function of the co-occurrence of suitable rainfall, receptive seedbed and seed (i.e. seed supply > losses due to ant theft or other causes). According to this model, the asymmetrical distributions of seedlings around parent trees would be solely due to wind direction at the time of seedfall. Proc. LINN. SOc. N.S.W., 119. 1998 152 EUCALYPT REGENERATION IN PASTURES The tendency for seedlings to be more common on the southern sides of trees — noted at Panuara and Mansfield, and also by Curtis (1989) — may have been due to dry northerly winds depositing more seed in the south, but in the absence of seed dispersal and wind data this hypothesis cannot be tested. Curtis (1989) concluded that best recruit- ment occurred just outside the canopy on the southern side of trees. In one experiment where eucalypt seed was sown (and hence, independent of wind-dispersed seed) in three locations: amongst trees, in the open and on the southern side of tree clumps, ‘the plots on the southern sides of the clump of trees had twice as many recruited seedlings than those directly under trees and six times as many as the open plots’ (p.108). He suggested that protection from frost and desiccation may have been responsible for this result. A higher density of seedlings on the southern sides and beneath the canopies of Eucalyptus pauciflora Sieber ex Sprengel trees was also noted in the Orroral Valley near Canberra. According to Egerton (1996), this could be explained by warmth provided by the canopy at night and protection from direct winter-morning sunlight, which can cause frost-induced photoinhibition in seedlings. Seedling deaths due to ‘frost heave’ have also been reported (Cremer 1990) in cold climates. However, frost effects were unlikely to explain seedling distribution at Panuara and Mansfield, where asymmetry was estab- lished well before frost damage could occur; and very few deaths were recorded over winter. Mid January shading at Panuara (Figure 6A) was not correlated with seedling num- bers. However, at Mansfield in mid February, increased shading in the south was evident (Figure 6B). Importantly, this pattern would be equivalent to that two months earlier, in mid December, when seedlings were first appearing. It is likely, therefore, that protection from the increasing levels of radiation of late spring/early summer allowed more seedlings to survive. Glasshouse trials by Curtis (1989) showed that increased frequency of watering of eucalypt seeds sown at shallow depth significantly increased emergence. Under field conditions, prolonged periods of wet weather would be expected to yield a similar result. Similarly, any factor, perhaps shading, which reduces the rate of topsoil drying would be expected to enhance emergence. Topsoil moisture data at Molong (Figure 7) suggested that the rate of drying was reduced on the eastern and southern sides of a tree. This may be critical to emergence in seasons when topsoil moisture is limiting. Woodland eucalypts in the subhumid zone of south-eastern Australia have been in decline for many years and lack of natural revegetation (due to grazing, sowing exotic pasture species and soil cultivation) has been cited (Nadolny 1995) as a major cause. These experiments have confirmed the importance of grazing (as evidenced by the lack of seedlings outside exclosures) and the presence of exotic pasture species as limiting factors. Some forms of cultivation, however, may enhance recruitment as also reported by Dalton (1993) and MacLennan et al. (1992). Seedfall from parent trees was not limit- ing in most cases but the importance of rainfall and climate modification afforded by the presence of existing trees in promoting recruitment has been highlighted. CONCLUSIONS The results of the experiments indicate that eucalypt recruitment in woodlands with non-native pastures can be enhanced by an appropriate seedbed, adequate seedfall (i.e. abundant fruits), stock exclusion and most importantly, above-average rainfall. An appropriate seedbed is one that has abundant bare ground, at least during early establishment, and some roughness in the form of vegetative residues. Both slashing/knockdown herbicide and scalping can satisfy these requirements, though the latter appears to need a ‘settling-in’ period (probably to allow some vegetative roughness to develop) to be fully receptive. Proc. LINN. SOC. N.S.W., 119. 1998 J. LAWRENCE, W.S. SEMPLE AND T.B. KOEN 153 Above-average rainfall in spring and December plus a follow-up of at least 80 mm in January, February or March appears to be required. Well above-average rainfall in the latter months can compensate for lower rainfall in spring but above-average rainfall in December appears to be critical. The limited seedfall data from one E. albens tree supported Curtis’ (1989) observa- tion of a summer seedfall peak in other woodland eucalypts. However, successful recruit- ment appears to be associated with the ‘rising limb’ of the seedfall graph rather than at the peak. Seedbed preparation as late as November enhanced natural regeneration in the two successful cases reported here. Limited evidence from these experiments and from other sources suggests that the area near the canopy on the southern side of trees is likely to be the site of maximum recruitment. Although this could be due to the presence of more wind-dispersed seed in this area, it is likely that environmental factors such as increased shading or enhanced soil moisture may be responsible. In favourable seasons, these factors may be of lesser importance. ACKNOWLEDGEMENTS Land for the experiments was provided by Messrs. J. Howie, “Maxwood Park”, Mansfield, B. Lorimer, “Weemalla”, Panuara and A. Evans, “Hillview Park”, Molong. The experiment at Mansfield was funded by the Victorian Government’s Sustainable Agriculture Strategy Project and the Murray-Darling Basin Commission’s Natural Resources Management Strategy, and those in NSW by the Soil Conservation Service, now part of the Department of Land and Water Conservation (DLWC). The skills of many individuals and groups were called upon between the initial planning and final manuscript stages of experiments reported above. The assistance of Mr. P. Haines, Mr. J. Maden and Mr. K. Wilson (all of Rutherglen), Mrs. E. Davidson, Dr. M. Campbell and Mr. R. Bath (all of Orange), Mr. D. Curtis (Armidale) and members of the Orange and Molong L.E.A.P. teams is gratefully acknowledged. Seed germination and soil moisture determinations were carried out by DLWC’s Research Centres at Cowra and Wellington respectively, and Figures 1 and 6B were prepared by DLWC’s G.L.S. Unit at Orange. REFERENCES Beckmann, R. and Davidson, S. (1990). Reversing tree decline. Rural Research 146, 19-26. Bird, P.R., Bicknell, D., Bulman, P.A., Burke, S.J.A., Leys, J., Parker, J.N., van der Sommen, F.J. and Voller, P. (1992). The role of shelter in Australia for protecting soils, plants and livestock. Agroforestry Systems 18, 59-86. Bird, P.R., Kearney, G.A. and Jowett, D.W. (1994). Trees and shrubs for south-west Victoria. Technical Report Series No. 205, Department of Agriculture, Victoria. Boland, D.J., Brooker, M.I-H., Turnbull, J.W. and Kleinig, D.A. (1980). ‘Eucalyptus seed’. (CSIRO: Canberra). Campbell, M.H. (1996). Establishment of surface-sown pastures on unploughed infertile soil. In “Proceedings eighth Australian agronomy conference, Toowoomba’ (Ed. M. Asghar) pp.116—-19. (Australian Society of Agronomy: Carlton). Campbell, M.H. and Nicol, H.I. (1996). Establishing trees on non-arable land to control weeds. In “Proceedings eleventh Australian weeds conference, Melbourne’ (Ed. R.C.H. Shepherd) pp. 493-496. (Weed Science Society of Victoria: Frankston). Campbell, R., Chandler, R. and Thomas, G. (1988). Trees and agriculture. In ‘Victoria felix: improving rural land with trees’ (Eds. A. Lindsay and R. Youl) pp.1—2. (Department of Conservation, Forests and Lands and Graduate School of Environmental Science Monash University: Victoria). Christensen, P.S. (1971). Stimulation of seedfall in karri. Australian Forestry 35, 182-190. Cluff, D. and Semple, W.S. (1994). Natural regeneration — in “Mother Nature’s” own time. Australian Journal of Soil and Water Conservation 7(4), 28-33. Cremer, K.W. (1990). Frost. In ‘Trees for rural Australia’ (Ed. K.W. Cremer) pp.217—224 (Inkata Press: Melbourne). Cremer, K.W., Unwin, G.K. and Tracey, J.G. (1990). Natural regeneration. In “Trees for rural Australia’ (Ed. K.W. Cremer) pp.107—129. (Inkata Press: Melbourne). Curtis, D. (1989). Eucalypt re-establishment on the Northern Tablelands of New South Wales. MSc thesis, University of New England, Armidale. Curtis, D. (1990). Natural regeneration of eucalypts in the New England region. In ‘Sowing the seeds. Direct seeding and regeneration conference’ pp.7—16. (Greening Australia: Deakin, ACT). Proc. LINN. Soc. N.S.W., 119. 1998 154 EUCALYPT REGENERATION IN PASTURES Dalton, G. (1993). “Direct seeding of trees and shrubs. A manual for Australian conditions’. (Primary Industries: Adelaide) Egerton, J. (1996). Tree planting in cold climates — lessons from fundamental research. Australian Journal of Soil and Water Conservation 9(1), 37-42. Florence, R.G. (1996). ‘Ecology and silviculture of eucalypt forests’. (CSIRO: Melbourne). Foster, M.B. (1985). Regeneration with native flora at Weipa. In “Proceedings of the North Australian mine rehabilitation workshop No. 9’ (Ed. J.W. Lawrie) pp.45—59. (Comalco Aluminium Ltd: Weipa, Qld). MacLennan, F., Robinson, J., Higgins, I. and Platt, S. (1992). Natural regeneration — case studies on the farm. Land for Wildlife Note No. 16, Department of Conservation and Environment, Victoria. Middleton, W.G.D. (1984). Maintenance and management of existing woodlands and plantations. In ‘Proceedings of the focus on farm trees conference, University of New England, Armidale, May 1984’ (Ed. J.A. Hofler) pp. 77—79. (Canberra Publishing and Printing Co.: Fyshwick). Nadolny, C. (1995). Causes of tree decline/dieback in NSW. In ‘Redressing rural tree decline in NSW’ (Ed. A. Kater) pp. 11-18. (Greening Australia NSW Inc.: Sydney). O’Dowd, D.J. and Gill, M. (1984). Predator satiation and site alteration following fire: mass reproduction of alpine ash (Eucalyptus delegatensis) in south eastern Australia. Ecology 65, 1052-1066. Semple, W.S. (1987). Prior streams and Queel Queel Gums: regeneration areas revisited. Journal of the Soil Conservation Service of NSW 43, 38-43. Semple, W.S. and Koen, T.B. (1997). Effect of seedbed on emergence and establishment from surface-sown and direct drilled seed of Eucalyptus spp. and Dodonaea viscosa. Rangeland Journal 19, 80-94. Venning, J. (1985). Natural regeneration: a South Australian perspective. Trees and Natural Resources 27(1), 9-11. Venning, J. (1988). ‘Growing trees for farms, parks and roadsides. A revegetation manual’ (Lothian: Melbourne). Setterfield, S.A. and Williams, R.J. (1996). Pattern of flowering and seed production in Eucalyptus miniata and E. tetrodonta in a tropical savanna woodland, northern Australia. Australian Journal of Botany 44, 107-122. Whalley, R.D.B. and Curtis, D.J. (1991). Natural regeneration of eucalyptus on grazing land on the Northern Tablelands of NSW, Australia. In ‘Proceedings of the [Vth international rangelands congress, Montpellier, France, April 1991’ (Eds. A. Gaston, M. Kernick, H. Le Houerou) pp.581—584. (Association Francaise de Pastoralisme: France). Yates, C.J., Hobbs, R.J. and Bell, R.W. (1996). Factors affecting the recruitment of Eucalyptus salmonophloia in remnant woodlands. III. Conditions necessary for seed germination. Australian Journal of Botany 44, 283-296. Proc. LINN. SOC. N.S.W., 119. 1998 A Complete Description of Sepia mira (Cotton 1932) (Cephalopoda: Sepiidae) from Eastern Australia AMANDA REID (Communicated by M.L. Augee) 140 Napoleon Street, Eltham, Victoria 3095 REID, A. (1998). A complete description of Sepia mira (Cotton 1932) (Cephalopoda: Sepiidae) from Eastern Australia. Proceedings of the Linnean Society of New South Wales 119, 155-171. Sepia mira (Cotton 1932) is described on the basis of five specimens trawled between 20-72 m off northern New South Wales. The species was known previously only from cuttle- bones collected from a number of eastern Australian localities. This complete description confirms the status of S. mira as valid. Manuscript received 20 July 1997, accepted for publication 17 September 1997. KEY WORDS: Sepia mira, Sepiidae, doratosepion, description, eastern Australia. INTRODUCTION Sepia mira was described by Cotton (1932) on the basis of a cuttlebone collected on North-West Islet in the Capricorn Group of Islands off Gladstone, Queensland. Since that time, bones have been collected from additional localities in New South Wales (NSW) and Queensland, but whole animals had remained elusive. In 1995 and 1996, Ken Graham from NSW Fisheries collected some representatives of this species near the Clarence River mouth in northern New South Wales, enabling the species to be fully described here. The specimens were collected as part of a NSW Fisheries survey of the Newcastle and Clarence River prawn grounds. Other sepiids collected within the range of S. mira were: S. apama Gray 1849; S. limata (Iredale 1926) (see endnote); S. opipara (Iredale 1926); S. plangon Gray 1849 and S. rozella (Iredale 1926), with the latter two species occurring in the greatest abundance (Graham and Wood 1997). Cotton (1932) decided that Sepia mira was sufficiently distinctive to place in a new genus, Tenuisepia. Subsequent workers (with the notable exception of Iredale and McMichael (1962) who placed the species in its own subfamily) have generally conclud- ed that distinct generic status is not justified. Until a complete revision of the sepiids is undertaken, the classification followed here is that proposed in Khromov et al. (in press) which recognises only three genera within the Sepiidae with the present species designat- ed to the genus Sepia. This complete description confirms the status of S. mira as valid. MATERIALS AND METHODS This work was based on museum material. All material studied is listed in the Material Examined section. Institutional acronyms used are: AM — Australian Museum, Sydney, Australia; MV — Museum of Victoria, Melbourne, Australia and SAM — South Australian Museum, Adelaide, Australia. Other abbreviations: coll. — collected, F — female, FV — Fisheries Vessel, Is. — Island, J — juvenile, m — meters, M — male, mm — millimeters. Proc. LINN. SOC. N.S.W., 119. 1998 156 COMPLETE DESCRIPTION OF SEPIA MIRA TABLE | Description of measurements and counts. Definitions largely follow Roper and Voss (1983). New or modified definitions are indicated by an asterisk (*). Indices (shown in square brackets) are calculated by dividing each measure by mantle length or, for cuttlebone characters, cuttlebone length (unless otherwise specified). Arm Length — AL: length of each designated (i.e. 1,2 etc.) arm measured from first basal (proximal-most) sucker to distal tip of arm (Arm 1, dorsal; 2, dorso-lateral; 3, ventro-lateral; 4, ventral) [ALI]. Anterior Mantle to Head length * — AMH: dorsal length of mantle measured from anterior-most point of man- tle to intersection of transverse line joining dorso-lateral mantle margin [AMHI]. Arm Sucker Count * — ASC: total number of suckers on each designated arm (e.g. ASC2). Arm Sucker diameter — AS: diameter of largest sucker on each designated (i.e. 1,2 etc.) arm [ASIn]. Suckers on left ventral hectocotylised arms are differentiated as follows: Arm Sucker left 4 * — ASI4: diameter of largest sucker on left ventral arm of male [ASInI4]. Cuttlebone Breadth — CbB: greatest dorso-ventral breadth of cuttlebone. [CbBI] Cuttlebone Length — CbL: dorsal length of cuttlebone along midline, including spine. Cuttlebone Width — CbW: greatest width of cuttlebone [CbWI]. Club Length — CIL: length of tentacular club measured from proximal-most basal suckers (carpus) to distal tip of club [CILT]. Club Row Count — CIRC: number of suckers in transverse rows on tentacular club. Club Sucker diameter — CIS: diameter of largest sucker on tentacular club [CISI]. Club Sucker dorsal * — ClSd: diameter of largest tentacular club sucker in dorsal-most (closest to swimming keel) longitudinal row [CISId]. Club Sucker ventral * — ClSv: diameter of largest tentacular club sucker in ventral-most (opposite swimming keel) longitudinal row [CISIv]. Eye Diameter — ED: diameter of eye [EDI]. Egg Length * — EgL: length of egg [EgLI]. Free Funnel length — FFu: the length of the funnel from the anterior funnel opening to the point of its dorsal attachment to the head [FFul]. Fin Insertion anterior * — Fla: anterior origin of fin measured from mantle margin to anterior-most junction of fin and mantle [FIIa]. Fin Insertion posterior* — FIp: measured between posterior junctions of fins with mantle [FIIp]. Funnel Length — FuL: the length of the funnel from the anterior funnel opening to the posterior margin mea- sured along the ventral midline [FuLI]. Fin Width — FW: greatest width of single fin [FWI]. Gill Lamellae Count — GiLC: number of lamellae on outer demibranch including the terminal lamella. Gill Length * — GiL: length of gill [GiLI]. Head Length — HL: dorsal length of head measured from point of fusion of dorsal arms to anterior tip of nuchal cartilage [HLI]. Head Width — HW: greatest width of head at level of eyes [HWI]. Loculus Length * — LoL: length of the last loculus (ventral anterior smooth zone of the cuttlebone) [LoL]. Mantle Length — ML: dorsal mantle length. Measured from anterior-most point of mantle to posterior apex of mantle. Mantle Width — MW: greatest straight-line ventral width of mantle [MWI]. Spine Length * — SL: length of spine [SLI]. Spermatophore Length — SpL: length of spermatophore [SpL]]. Spermatophore Width — SpW: greatest width of spermatophore. Spermatophore width index is expressed as a percentage of spermatophore length [SpWI]. Striated Zone length — StZ: length of striated zone of cuttlebone [StZI]. Transverse Row Count — TrRC: number of suckers in longitudinal series on tentacular club [counted from proximal-most basal suckers (carpus) to distal tip of club]. Ventral Mantle Length — VML: length of ventral mantle measured from anterior mantle margin at ventral midline, to posterior apex of mantle [VMLI]. Proc. LINN. SOC. N.S.W., 119. 1998 H ie C H Cc i 7 d Ww meersea KOE ZENS OEE Ro spaces =iNFe R L1L2M ey) Oy Figure 1. Measurements and Terminology: (a) whole animal dorsal view (for abbreviations and definitions see Table 1); (b) tentacular club (C — carpus, D — dorsal, PM — protective membranes, SK — swimming keel, V- ventral). The number of suckers intersected in an oblique transverse line across the club, shown as a hatched line on this figure, is the Club Row Count (CIRC) [In the example illustrated CIRC = 4]; (c) upper beak (C — crest, H — hood, R — rostrum); (d) lower beak (C — crest, H — hood, L — lateral wall, W — wing); (e) arm sucker rim (I — inner ring, INF — infundibulum, PO — polygonal process, P — peg); (f) radula (R — rhachidian teeth, L1 — first lateral teeth, L2 — second lateral teeth, M — marginal teeth); (g) cuttlebone, ventral view (for abbreviations and definitions, see Table 1). [(a) and (g) modified from Roper and Voss (1983) Figure 1]. 158 COMPLETE DESCRIPTION OF SEPIA MIRA TABLE 2 Measurements (mm), counts and indices of 3 male and 2 female Sepia mira (Cotton). M — male, F — female. Museum MV MV AM MV AM Reg. No. F80995 F80996 C306764 F80997 C306387 M M M F F ML 38.1 42.7 44.0 43.0 43.9 MWI 43.8 39.3 43.2 46.7 45.6 AMHI 11.0 12.6 WA3} 9.3 10.9 VMLI 85.3 84.3 83.6 90.0 80.6 FWI 6.3 8.4 8.4 9.3 les Fila 6.3 2.8 Doi Doll 4.6 FIIp 4.7 33) 4.5 D333} 9.6 FuLlI 26.1 29.5 Do 29.1 28.5 FFul 11.8 10.5 14.8 93 9.1 HLI 27.1 23.0 22.3 22.6 28.5 HWI BES 32.3 30.7 34.7 31.9 EDI 12.6 10.8 10.5 10.5 8.0 ALI1 31.5 31.6 25.0 34.9 DY) ALI2 30.2 ; 29.3 36.4 32.6 31.9 ALI3 BIE 25.8 31.8 31.4 37.6 ALI4 39.4 SD 38.6 39.5 36.4 ASIn1 0.84 1.03 1.02 LW 1.03 ASIn2 1.05 1.03 1.14 1.05 1.03 ASIn3 1.05 1.17 1.25 1.12 es) ASIn4 1.18 0.94 1.18 1.28 I 725) ASC1 80 78 90 90 82 ASC2 60 82 95 92 82 ASC3 60 78 110 103 98 ASC4 92 122 104 116 110 ASIn14 1.05 0.70 1.14 - - CILI 11.8 ED) 10.0 10.0 - CIRC 3 3 3 3 - TrRC 16 13 17 20 - CISI 1.34 1.41 1.36 0.93 - ClISId 0.63 0.63 0.80 0.56 ~ CISIv 0.39 0.35 0.34 0.37 - GiLC - 26 - 24 - GiLI 28.3 28.3 34.1 30.0 27.3 SpLI 7.9 6.6 6.6 - = SpWI 33 Savill 4.14 - - EgDI ~ ~ - 5.8 V3) CbL 39.9 47.9 44.0 51.4 45.1 CbWI 22.8 77h D2) D3) 22.0 CbBI 7.0 7.3 7.9 5.8 V3 SLI 3.9 3.4 Tf 4.5 3.3 StZ] 69.6 63.9 66.6 72.8 67.2 LoLI 21.0 28.8 30.5 20.2 30.8 LoL/StZ (%) 30.2 45.1 45.7 Die 45.9 Proc. LINN. SOc. N.S.W., 119. 1998 A. REID 159 TABLE 3 Sepia mira (Cotton); ranges of arm length indices (ALI), arm sucker diameter indices (ASIn) and arm sucker counts (ASC) of 3 mature males and 2 mature females. min. = minimum, max. = maximum, SD = standard deviation. Males Females min. mean max. SD min. mean max. SD ALI1 25.0 29.4 31.6 3.8 DATES aiilail 34.9 4 a3 ALI2 29.3 Be9 36.4 32) 31.9 32 32.6 0.5 ALI3 25.8 ONT 31.8 3.4 31.4 34.5 37.6 4.4 ALI4 37.5 38.5 39.4 1.0 36.4 38.0 3915 De, ASIn1 0.84 0.96 1.03 0.11 1.03 1.07 Les 0.06 ASIn2 1.03 1.07 1.14 0.06 1.03 1.04 1.05 0.02 ASIn3 1.05 1.16 1.25 0.10 le 1.18 EDS 0.10 ASIn4 0.94 1.10 1.18 0.14 1.25 eT 1.28 0.02 ASC1 78 83 90 6 82 86 90 6 ASC2 60 19) 95 18 82 87 92 7 ASC3 60 83 110 7S) 98 101 103 4 106 122 15 110 113 116 4 ASC4 92 Measurements and indices used throughout this paper are primarily those given in Roper and Voss (1983), using dorsal mantle length (ML) as a size standard. Some addi- tional measurements are used, and these with the definitions listed by Roper and Voss (1983) are given in Table 1. Parts of the club and arm sucker rims are described using the terminology of Nixon and Dilly (1977), while nomenclature for the radula follows Nixon (1995). Beaks were described following Clarke (1986). Diagrammatic illustrations of measurements and terminology used for key structures are shown in Fig. 1. Measurements were made either using dial callipers, or an eyepiece micrometer attached to a stereo microscope. All measurements are expressed in millimetres (mm). Measurements and counts for individual specimens are shown in Table 2. Ranges of arm length indices, arm sucker diameter indices, and arm sucker counts are presented in Table 3; ranges for all other characters appear in the text. In species descriptions and tables, the range of values for each character is expressed as: minimum — mean — max- imum (standard deviation, SD). Values for each sex are given separately. Measurements for structures which were clearly distorted or broken were not attempted, and these, in addition to missing values, appear as a dash (—) in the tables. Ranges for specific character traits given with the species description do not, therefore, always refer to the total number of specimens examined. For examination of arm and club sucker rims, suckers were removed from the mid- dle of designated arms and the tentacular club, mounted in glycerine jelly and viewed using a compound microscope. Radulae and beaks were dissected from the buccal mass, and soaked for approximately 30 minutes in a warm, saturated potassium hydroxide solu- tion, then radulae were cleaned using forceps and a fine brush. Radulae were also mount- ed in glycerine jelly, and the new, unused portion was examined. The species description was generated by the DELTA (Description Language for Taxonomy) system (Dallwitz 1980; Dallwitz et al. 1993; Partridge et al. 1993). Proc. LINN. Soc. N.S.W., 119. 1998 160 COMPLETE DESCRIPTION OF SEPIA MIRA Sepia mira (Cotton 1932) (Figures 2-9, Tables 2 & 3) Synonymy Tenuisepia mira Cotton 1932: 546-547, figs 7—9.-Iredale 1954: 75, pl. V, figs 7 & 8.—Iredale & McMichael 1962: 99. Sepia mira.— Adam & Rees 1966: 87.—Nesis 1982: 119, fig. 24X.—Lu & Phillips 1985225: Material examined Holotype: Australia: Queensland, North-West Is. — cuttlebone (55 mm CbL), 23°18'S, 151°42’E, coll. W.J. Kimber, (SAM D10507). Additional Material: Australia: Queensland — 5 cuttlebones (41.5—42.5 CbL), Lizard Is., Coconut Beach, 14°40’S 145°28’E, 10 Oct 1982, coll. R. Burn, (MV F80999); cuttlebone (37.3 CbL), Heron Is., 23°26’'S 151°55’E, 1988, coll. A. Reid & P. Ward, (MV F80998). New South Wales — 1F (43.0 mm ML), Clarence R., 29°19’S 153°29’E — 29°20'S 153°29'E, 49-48 m, 19 Apr 1996, coll. K. Graham on FV “Kapala” K960638, (MV F80997); 1M (44.0 mm ML), Off Iluka, 29°20’S 153°25’E — 29°19'S 153°25’E, 37-34 m, 29 May 1995, coll. K.J. Graham on FV “Kapala” K950620, (AM C306764); 1M (42.7 mm ML), Clarence R., 29°22'S 153°23’E — 29°23'S 153°23’E, 28-20 m, 31 Aug 1995, coll. K. Graham on FV “Kapala” K951131, (MV F80996); 1M (38.1 mm ML), Clarence R., 29°50’S 153°20'E — 29°48'S 153°20’E, 37-33 m, 10 Apr 1996, coll. K. Graham on FV “Kapala’, K960602 (MV F80995); 1F (43.9 mm ML), off Wooli, 29°49’'S 153°27'E — 29°48'S 153°27’E, 72-69 m, 18 May 1995, coll. K. Graham on FV “Kapala’”, K950604 (AM C306387); cuttlebone (48 mm CbL), Trial Bay, 30°53’S 153°04’E, coll. M. Ward, (AM C133312). Diagnosis Cuttlebone oblong; spine with ventral keel; striated zone convex; anterior striae slightly convex to straight; sulcus very shallow, narrow; last loculus very short; inner cone limbs straplike anteriorly, narrower, thicker posteriorly; outer cone posterior lateral limbs flared ventro-laterally. Arm suckers tetraserial. Tentacular club suckers differ in size, 3-4 enlarged suckers toward posterior end of club; 3 suckers in transverse rows; dorsal and ventral protective membranes not fused at base of club; swimming keel extends beyond carpus. [Modified from the original description by Cotton (1932, p. 546)]. Description Counts and indices for individual specimens are given in Table 2; ranges for arm length indices, arm sucker diameter indices and arm sucker counts are shown in Table 3. Small to moderate-sized species (Fig. 2); ML males 38.1—41.6—44.0 (SD, 3.1), females 43.0-43.5—43.9 (SD, 0.6). Mantle oblong; MWI males 39.3—42.1—43.8 (SD, 2.4), females 45.6-46.2-46.7 (SD, 0.8); dorsal anterior margin triangular, acute; extend- ing to level of posterior margin of eyes; AMHI males 11.0—12.0—12.6 (SD, 0.8), females 9.3-10.1-10.9 (SD, 1.2). Ventral mantle margin weakly convex; VMLI males 83.6-84.4-85.3 (SD,0.8), females 80.6-85.3-90.0 (SD, 6.6). Fins widest in posterior third; FWI males 6.3—7.7-8.4 (SD,1.2), females 7.5—8.4—9.3 (SD,1.3); anterior origin posterior to mantle margin; Flla males 2.8-4.9-6.3 (SD, 1.9), females 4.6-4.9-5.1 (SD, 0.3); rounded posteriorly; narrow gap between fins; FIIp males 3.3-4.2-4.7 (SD, 0.8), females 9.6—L0.9-12.3 (SD, 2.0). Funnel long, broad-based; extends to level of anterior Proc. LINN. Soc. N.S.W., 119. 1998 A. REID 161 Figure 2. Sepia mira: left, cuttlebone dorsal view (posterior end at top); right, freshly caught animal dorsal view, cuttlebone removed, male MV F80996, 42.7 mm ML [photo K. Graham]. rim of eye; FuLI males 25.7—27.1—29.5 (SD, 2.1), females 28.5—28.8—29.1 (SD, 0.4). Funnel free portion approximately one-third funnel length; FFul males 10.5—12.4—14.8 (SD, 2.2), females 9.1—9.2—9.3 (SD, 0.1). Funnel organ dorsal elements inverted V-shape with small anterior papilla; ventral elements oval (Fig. 3a). Mantle-locking cartilage curved, with semicircular ridge; funnel-locking cartilage with depression which corre- sponds to ridge (Fig. 3b). Head short (Fig. 2); HLI males 22.3—24.1—27.1 (SD, 2.6), females 22.6—25.5—28.5 (SD, 4.2); slender, narrower than mantle; HWI males 30.7—31.5—32.3 (SD, 0.8), females 31.9-33.3-34.7 (SD, 2.0). Eyes moderate size; EDI males 10.5—11.3—12.6 (SD, 1.2), females 8.0—9.2-10.5 (SD, 1.8); ventral eyelids present (anterior half of eye only). Male arms 4 slightly longer than other arms, arms 1—3 all similar length, not elon- gate (Table 3). Female arm lengths subequal (Table 3). ALI of longest arms in males (ALI4) 37.5-38.1—38.6 (SD, 0.8), females (ALI4) 36.4-38.0—-39.5 (SD, 2.2). Protective membranes (both sexes) narrow; normal, not thickened. Distal arm tips (both sexes) not markedly attenuate. Arm suckers tetraserial in both sexes. Suckers in males normal in size (not greatly enlarged); similar to female arm suckers in size (Table 3). Chitinous Proc. LINN. SOC. N.S.W., 119. 1998 162 COMPLETE DESCRIPTION OF SEPIA MIRA yn WI UT Maree Vaan ausnssees © a OVS ®: Figure 3. Sepia mira: (a) funnel organ, male, MV F80996, 42.7 mm ML, scale bar 2 mm; (b) funnel locking cartilage (left), and mantle locking cartilage (right), female, MV F80997, 43 mm ML, scale bar 2 mm; (c) suck- er rim arm 2, scale bar 50 wm, arrow indicates elongate teeth on distal margin of inner ring; (d) enlargement of sucker rim arm 2, scale bar 50 ym; (e) hectocotylus, left arm 4, scale bar 2 mm; (f) right arm 4, scale bar 2 mm. (a-b & e-f, male MV F80995, 38.1 mm ML). rims of arm suckers with elongate rectangular teeth on distal half of inner ring, teeth much smaller on proximal half of ring; infundibulum with 4-5 rows of hexagonal processes, inner 2—3 rows with elongate rounded pegs on distal half, becoming smaller towards periphery of sucker, shorter pegs on proximal half as for inner ring; peripheral sucker rim processes radially arranged, elongate, without pegs (Figs 3c and 3d). Sucker counts range from 60—122; females with higher average counts than males (Table 3). Hectocotylus present, left ventral arm modified (see remarks); sucker size slightly reduced proximally; suckers equal in size across rows, maximum sucker diameters: ASInI4 0.70—0.96—1.14 (SD, 0.23) (compare with right ventral arm, Table 3). Oral sur- face of modified region wide, fleshy, with transversely grooved ridges (Fig. 3e, compare with Fig. 3f); without distinct median furrow. Proc. LINN. SOC. N.S.W., 119. 1998 A. REID 163 Figure 4. Sepia mira: (a) tentacular club, oral view; (b) tentacular club, oral view, dorsal protective membrane pinned back to show sucker arrangement; (c) tentacular club, aboral view. (a-c, female, MV F80997, 43.0 mm ML, scale bars 2 mm). Tentacular club similar length in males and females; CILI males 10.0—11.0-11.8 (SD, 0.9), female 10. Club crescent-shaped; sucker-bearing face flattened (in all speci- mens examined, posterior half to two thirds of the club is folded toward dorsal side, ven- tral protective membrane covering posterior suckers, Fig. 4a). Club with 3 suckers in transverse rows in both sexes; 13-20 suckers in longitudinal series, TrRC males 13-15-17 (SD, 2), female 20. Suckers differ markedly in size, 3—4 enlarged suckers towards posterior end of club (Fig. 4b); CISI males 1.34—-1.37—1.41 (SD, 0.03), female 0.93; dorsal and ventral marginal longitudinal series of suckers differ slightly in size; dorsal marginal longitudinal series of suckers slightly larger than those in ventral margin- al series; CISId males 0.63—0.69—0.80 (SD, 0.09), female 0.56; CISIv males 0.34—0.36—-0.39 (SD, 0.03), female 0.37. Sucker dentition: half inner ring circumference in both sexes with elongate rectangular teeth, remaining half with blunt projections; infundibulum with 3-4 rows of hexagonal processes, innermost with elongate rounded pegs, pegs smaller towards periphery of sucker; at periphery, processes smaller, elongate- rectangular, without pegs (Figs 5a and 5b) (similar to arm suckers). Swimming keel of club extends well beyond carpus. Dorsal and ventral protective membranes not fused at base of club; joined to stalk; dorsal and ventral membranes same length, terminate at posterior end of carpus (Figs 4a and 4b); approximately equal width (Fig. 4c); dorsal membrane forms shallow cleft at junction with stalk. Gills with 24—26 lamellae per demibranch; GiLC in only intact male 26, 24 in only intact female. Gill length: GiLI males 28.3—30.3—34.1 (SD, 3.3), females 27.3—28.7—30.0 (SIDII29)): Buccal membrane without suckers. Upper beak (Fig. 5c) rostrum sharply pointed, long, length greater than width, cutting edge slightly curved; hood high above crest poste- riorly; crest curved, lateral wall shallowly indented posteriorly; wings narrow and short; jaw angle approximately 90 degrees, slightly acute; hood dark brown, lighter on ventral Proc. LINN. Soc. N.S.W., 119. 1998 164 COMPLETE DESCRIPTION OF SEPIA MIRA /\ Figure 5. Sepia mira: (a) club sucker rim, large club sucker, scale bar 100 ym; (b) enlargements of large club sucker rim, scale bar 50 sm; (c) upper beak, side view; (d) lower beak, side view; (e) lower beak, ventral view; (f) radula, male, MV F80996, 42.7 mm ML, scale bar 200 um; (g) digestive tract, dorsal view, scale bar 500 um (A — anus; BM — buccal mass; C — caecum; DG — digestive gland; IS — ink sac; P — pancreas; S — stomach; SG — salivary gland). (a—b & g, male MV F80995, 38.1 mm ML; c-e, female, MV F80997, 43.0 mm ML, scale bars 3 mm). margin, crest only slightly pigmented. Lower beak (Fig. 5d) rostrum protrudes only slight- ly, cutting edge curved; hood low on crest; crest straight, no indentation on lateral wall edge; hood and wings, width narrow; hood notch deep, broad (Fig. 5e); wings widely spaced; crest wide; rostrum pigmented dark brown, wings and crest only slightly pigment- ed. Radula homodont; rhachidian teeth with narrow, truncate bases, slender, triangular, sides slightly concave (Fig. 5f); first lateral teeth similar length and width to rhachidian teeth, asymmetrical with mesocone slightly displaced toward centre of radula; second lat- erals longer than first, distinctly curved on lateral margin, with broad heels; marginal teeth elongate with long tapered and curved mesocone (Fig. 4f). Digestive tract: (Fig. 5g) paired salivary glands approximately one-third length of buccal mass; paired digestive Proc. LINN. SOC. N.S.W., 119. 1998 A. REID 165 Figure 6. Sepia mira: (a) male reproductive tract (testis not shown; spermatophore storage sac containing sper- matophores), scale bar 2 mm (AAG — appendix of accessory gland; AG — accessory gland; CC — ciliated canal; DDC — distal deferent canal; GO — genital orifice, MG — mucilaginous gland; SG — spermatophoric gland; SSS — spermatophore storage sac; VD — vas deferens); (b) spermatophore, scale bar 300 yam; (c) spermatophore, oral end, scale bar 100 yym; (d) female reproductive tract MV F80997, 43.0 mm ML, scale bar 3 mm (ANG — accessory nida- mental gland; GO — genital opening; NG — nidamental gland; O — ovary). (a-c male MV F80995, 38.1 mm ML). Proc. LINN. Soc. N.S.W., 119. 1998 166 COMPLETE DESCRIPTION OF SEPIA MIRA Figure 7. Sepia mira: (a) cuttlebone, dorsal view, holotype D10507, 55.0 mm CbL, scale bar | cm; (b) cuttle- bone, ventral view, same specimen, scale bar 1 cm [photo C. Rowley]; (c) posterior end of cuttlebone, ventral view, AM C306764, 44.0 mm CbL, scale bar 2mm. Proc. LINN. SOC. N.S.W., 119. 1998 A. REID 167 glands large, located close together, with narrow, elongate triangular lobes posteriorly, ducts (not shown in figure) connect digestive glands near midline with caecum, ducts with branched attached pancreatic tissue; oesophagus runs dorsally along median junction of digestive glands, joins sac-like stomach immediately posterior to digestive glands; caecum disc-like, grooved in a blunt V-shape anteriorly, surface lining finely pleated; intestine undifferentiated; ink sac very large, elongate; anal flaps well developed. Male reproductive tract: testis on left posterior side of viscero-pericardial coelom; at distal end, convoluted vas deferens opens into broad, cone-shaped mucilaginous gland, then narrower, curved, spermatophoric gland (Fig. 6a). Close to junction with lobe- shaped accessory gland and gland appendix, delicate ciliated canal joins spermatophoric gland; distal deferent canal connects appendix of accessory gland to spermatophore stor- age sac; genital orifice opens dorsal to left gill in anterior end of mantle cavity. Spermatophores: cement body bipartite (Figs 6b and 6c); aboral end rounded, bulbous, connects to sperm reservoir via narrow duct; oral end of cement body cone-shaped, approximately twice length of, and slightly narrower than aboral end, connects to aboral end via long neck, tapers toward oral extremity of cement body; middle tunic com- mences along aboral part of cement body; ejaculatory apparatus coiled, extends into oral dilation of spermatophore. Spermatophores 2.8—3.0 mm long, 0.12—0.16 mm wide; SpLI 6.6-7.0—7.9 (SD, 0.8); SpWI 4.14—5.06—5.71 (SD, 0.82). Smallest male with well devel- oped spermatophores in Needham’s sac is 38.1 mm ML. Female reproductive tract: ovary hangs medially from dorsal wall of posterior vis- cero pericardial coelom. Oviduct thin-walled, continuous with body cavity; distally with thickened, glandular walls (oviducal glands). Nidamental glands in mature animals occu- py large portion of ventral side of mantle cavity. Accessory nidamental glands anterior to nidamental glands (Fig. 6d). Eggs oval; 2.5—-3.2 mm long; EgLI 5.8—6.6—7.3 (SD, 1.0). Smallest female with eggs in ovaries is 43.0 mm ML. Subdermal cartilaginous layer between cuttlebone and skin absent. Cuttlebone length approximately equal to mantle length; outline oblong (Figs 2, 7 and 8); CbL males 39.9-43.9-47.9 (SD, 4.0), females 45.1-48.3-51.4 (SD, 4.5); CbWI males 22.1-22.6—22.8 (SD, 0.4), females 22.0-22.8—23.7 (SD, 1.3); not strongly convex in lat- eral view; CbBI males 7.0—7.1—7.3 (SD, 0.2), females 5.8—7.0—7.0 (SD, 1.1). Bone blunt- ly rounded anteriorly; bluntly rounded posteriorly; not strongly recurved ventrally. Dorsal surface pinkish laterally (see remarks); evenly convex; granulose, calcified dor- sally, particularly posteriorly (not as pronounced on flared outer cone). Dorsal median rib absent; lateral ribs absent. Chitin borders lateral and anterior margins of cuttlebone. Spine present, long; SLI males 2.7—3.3—3.6 (SD, 0.5), females 3.3—3.6—3.9 (SD, 0.4); straight, directed dorsally; with ventral keel (Fig. 8b); fine, radiating ribs between outer cone and spine (Fig. 7c). Dorso-posterior end of cuttlebone without median longitudinal ridge anterior to spine. Striated zone convex (with narrow, flat margins, Fig. 8b); StZI males 63.9-66.7—69.6 (SD, 2.9), females 67.2—70.0—72.8 (SD, 3.9). Last loculus convex (strongly convex immediately anterior to striated zone, becoming flat posteriorly); LoLI males 21.0—26.8—30.5 (SD, 5.0), females 20.2—25.5—30.8 (SD, 7.5); at midline half length of striated zone (approximately), LoL/StZ(%) males 30.2—-40.3-45.7 (SD, 8.8), females 27.1—36.5—45.9 (SD, 13.3); loculus extends posteriorly as narrow margin on each side of striated zone. Sulcus extends entire length of cuttlebone; shallow, narrow (indistinct, visible only in large specimens); not flanked by rounded ribs. Last loculus with shallow median indentation, not very pronounced. Anterior striae broad inverted U- shape to straight (Fig. 8a). Limbs of inner cone extend anteriorly to approximately halfway along striated zone. Inner cone lateral limbs overlie calcareous striated zone anteriorly, extreme anterior tips bordered, separated from outer cone by striated zone. Inner cone limbs strap-like anteriorly, narrower posteriorly; raised, separated from striat- ed zone to form rounded ledge posteriorly; thickened (Fig. 7c). Inner cone without irreg- ular calcareous ribs posteriorly. Outer cone calcified; narrow anteriorly, broadens posteri- Proc. LINN. SOc. N.S.W., 119. 1998 168 COMPLETE DESCRIPTION OF SEPIA MIRA a Rtas i at, eh ans fe 4 = Figure 8. Sepia mira: (a) cuttlebone, ventral view, MV F80998, 37.3 mm CbL, scale bar 5 mm; (b) posterior end of cuttlebone, ventral view, same specimen, scale bar 3 mm. orly; lateral limbs flared ventro-laterally; limbs continue as narrow ledge ventral to spine (swollen slightly to cover posterior end of inner cone). Body papillae present (visible on a single specimen only); dorsal mantle with scat- tered short longitudinal ridges concentrated mid-dorsally and in a single row close to fins; ventral mantle without ridges. Head and arm papillae absent. Colour: head, arms and dor- sal mantle reddish brown (some dull orange spots below chromatophores); paired dorsal eye spots absent. Fins pale; without markings at base. Body without ventral pigment. Distinctive pigment spots on ventral side of club, spots in radiating rows extending from central region towards ventral margin (Fig. 4c). Mantle ridges orange-pink in colour. Type locality Queensland, North-West Islet, Capricorn Group, 23°18'S 151°42’E. Proc. LINN. SOC. N.S.W., 119. 1998 A. REID 169 Type Holotype by original designation. SAM: D10507. Cuttlebone only, 55 mm CbL. Distribution Queensland, Lizard Is., 14°40'S 145°28’E — Trial Bay, 30°53’S 153°04’E (cuttle- bones); New South Wales, Clarence R., 29°19’S 153°29'E — off Wooli, 29°49'S 153°27’E (entire animals) (Fig. 9); depth range 72—20 m. Remarks Contrary to Cotton’s (1932) original description, a very faint sulcus was seen in large specimens examined in this study. As Cotton (1932) described, the sulcus cannot be seen on the type specimen (Fig. 7b), possibly due to wear on the ventral surface of this beach-collected cuttlebone. In other respects, the cuttkebones examined here do not differ from the original description. Adam and Rees (1966) report the absence of a keel on the spine, but a ventral keel is clearly present in this species. The dorso-lateral sides of the cuttlebone are pinkish in freshly preserved animals, but this colour is lost in specimens that have been stored in ethanol. Of the three male specimens collected, the spermatophores were well developed in one specimen only (MV F80995). The modification of the left ventral arm could be seen only in this specimen. The arms of this animal are not in very good condition, so the details of the modification of the hectocotylus needs to be confirmed when more material is available. In particular, as a number of suckers are missing from this specimen, it is difficult to determine the relative sizes of the suckers in the modified region. It is not possible to ascertain whether the folding of the posterior half of the tentac- ular club, seen in all specimens, is an artefact of preservation, or if this is characteristic for the species. It has not been observed in other species of sepiids examined by the author. The five animals upon which the above description is based were collected from a narrow geographic range off the northeastern coast of New South Wales (Fig. 9). Cuttlebones have been collected at considerable distances north of this region, suggest- ing that the distribution of this species may be more extensive than that indicated from the complete specimens, though the bones may have drifted some distance to these loca- tions. The small size of the species possibly accounts for the absence of material from museum collections until now. It may be escaping trawl nets, or be of insufficient interest to most collectors to be retained. Determination of the relationships of Sepia mira to other species is at present diffi- cult owing to the general lack of knowledge about the phylogeny of the sepiids. Khromoy et al. (in press) have suggested that S$. mira belongs to the ‘doratosepion’ species complex. This name is based on Rochebrune’s (1884) generic name Doratosepion, though the validity of this genus has generally been rejected. The ‘doratosepion’ species complex (which may or may not be monophyletic) includes those species with an elongated body, short arms with biserial suckers, short tentacular clubs with unequal suckers, and a narrow elongate cuttlebone with two posterior ‘wings’ and a spine (Roeleveld 1972). In addition to the presence of biserial suckers on some or all of the arms, doratosepia often have modified arms in one or both sexes, some pairs may be elongate, tapering to a very narrow threadlike tip, or differ in sucker arrangement to the remaining arms. Most members of the group show sexual dimorphism. While sharing with members of this complex an elongated body, narrow bone, U-shaped inner cone, and unequal club suckers, S. mira does not have any distinctive modification of the arms (though they are short) or arm suckers, and little sexual dimorphism is evident. The outer Proc. LINN. Soc. N.S.W., 119. 1998 170 COMPLETE DESCRIPTION OF SEPIA MIRA Figure 9. Distribution of Sepia mira. Triangles indicate collection localities for cuttlebones only, circles indi- cate whole animals. The open triangle shows the type locality. cone in S. mira has been described as consisting of two long ‘wings’ (Adam and Rees 1966), though, as these authors acknowledge, they do not form a recurved ‘cup-like’ expansion as is often seen in other narrow-boned species. The outer cone ‘wings’ are usually shorter in these species than in S. mira, though the outer cone ‘wings’ of S. elon- gata d’Orbigny 1845 from the Red Sea (figured in Adam and Rees (1966), PI. 21, Fig. 132) are not dissimilar in appearance to those of S. mira in ventral view. Many species with broad cuttlebones, and therefore not included within the ‘doratosepion’ species complex, also have a ventro-laterally flared posterior outer cone. According to Cotton (1932), S. mira shows some affinity to a species described by Iredale (1926) as Decorisepia, type species S. rex. Though not stated in the original description, the reason for this may be that S$. mira and S. rex both have a very narrow inner cone and, were thought to lack a sulcus (though a sulcus is present in both species). Proc. LINN. SOC. N.S.W., 119. 1998 A. REID 17] Sepia rex differs greatly from S. mira in many other respects; the bone is much wider, has dorsal ribs and differs in shape, the club suckers are all small and arranged in 10—12 transverse rows, and the animal is much larger, ranging up to approximately 10 cm ML. It is unlikely that the two species share close affinities. Until we develop some working hypotheses of the phylogenetic relationships among the sepiids, the position of S. mira and particularly its association with other nar- row-boned species remains equivocal. ACKNOWLEDGEMENTS I wish to thank Ken Graham from NSW Fisheries for his interest in collecting and retaining the Sepia mira specimens described here, and Ian Loch (AM) for the loan of some of this material. I thank Ken Graham also for providing the photograph of the freshly collected animal and C.C. Lu for the photographs of the cuttle- bone of the holotype. REFERENCES Adam, W. and Rees, W.J. (1966). A review of the cephalopod family Septidae. Scientific Reports of the John Murray Expedition 1933-1934 11(1), 1-165, 46 plates. Cotton, B.C. (1932). Notes on Australian Mollusca with descriptions of new genera and species. Records of the South. Australian Museum 4(4), 537-547. Clarke, M.R. (1986). “Handbook for the Identification of Cephalopod Beaks’. (Oxford Science Publications: New York). Dallwitz, M.J. (1980). A general system for coding taxonomic descriptions. Taxon 29, 41-46. Dallwitz, M.J., Paine, T.A. and Zurcher, E.J. (1993). ‘User’s Guide to the DELTA System: a General System for Processing Taxonomic Descriptions.’ 4th edition. 136 pp. (CSIRO Division of Entomology: Canberra). Graham, K.J. and Wood, B.R. (1997). The 1995-96 survey of Newcastle and Clarence River prawn grounds. Kapala Cruise Report No. 116. (NSW Fisheries: Australia). Iredale, T. (1926). The cuttle-fish “bones” of the Sydney beaches. Australian Zoologist. 4: 186-196. Iredale, T. (1954). Cuttle-fish “bones” again. Australian Zoologist. 12(1): 63-82. Iredale, T. and McMichael, D.F. (1962). A reference list of the marine mollusca of New South Wales. Memoirs of the Australian Museum. 11, 1-109. Khromov, D.N., Lu, C.C., Guerra, A., Dong, Zh. and Boletzky, S.V. (in press). A synopsis of Sepiidae outside Australian waters (Cephalopoda: Sepioidea). Smithsonian Contributions to Zoology. Lu, C.C. (in press). A synopsis of Sepiidae in Australian waters (Sepioidea: Cephalopoda). Smithsonian Contributions to Zoology. Lu, C.C. and Phillips, J.U. (1985). An annotated checklist of the Cephalopoda from Australian waters. Occasional Papers. Museum of Victoria 2: 21-36. Nesis, K.N. (1982). “Cephalopods of the world’. (T.F.H. Publications Inc. Ltd : Neptune City, New Jersey) 351 pp. (Levitov, B.S. trans. from Russian, Burgess, L.A. ed. 1987). Nixon, M. (1995). A nomenclature for the radula of the Cephalopoda (Mollusca) — living and fossil. Journal of Zoology (London) 236, 73-81. Nixon, M. and Dilly, P.N. (1977). Sucker surfaces and prey capture. In: ‘The Biology of Cephalopods’. (Eds M. Nixon and J.B. Messenger) pp. 447-511. Symposium of the Zoological Society, London 38. Partridge, T.R., Dallwitz, M.J. and Watson, L. (1993). ‘A Primer for the DELTA System.’ 3rd edition. 15 pp. (CSIRO Division of Entomology: Canberra). Rochebrune, A.T. de (1884). Etude Monographique de la Famille des Sepiadae. Bulletin des Sciences, par la Société philomathique, Paris 7(8), 7-44, plates 3-6. Roeleveld, M.A. (1972). A review of the Sepiidae of southern Africa. Annales of the South African Museum 59(10): 193-313. Roper, C.F.E. and Voss, G.L. (1983). Guidelines for taxonomic descriptions of cephalopod species. Memoirs of the National Museum of Victoria 44, 48-63. ENDNOTE Sepia limata was misidentified as S. braggi Verco 1907 in Graham and Wood (1997), based on the work of Lu (in press) that includes S. Jimata as a synonym of S. braggi. Work in progress and in preparation for pub- lication by A. Reid has shown S. limata to be valid and distinct from S. braggi. Proc. LINN. SOc. N.S.W., 119. 1998 wes ate i D i20:Tr, yi ‘44 bls ct Fires th enbr il a4 PS ‘eae RT } caper he ; ute 7 " aa y te aS 7 tA : nw . ihe tices”) der Paine. OL i) RY WY! ; yeah? anh 2h AS Ef 4 : : } ra =u Pa a> <@ wea VS Sy Vel wad, seh aon vt nA hee we ML LeASELP rz ad b Le aie aye AST a a ae ie id OCT) als ass _ 7 x. sts Laue is VI c i in ets rr. i athe, ay or vc ae 7 Med ie ime WH ue Pace), ah upid hie male bi ¥iA ee 7 S a 7 2 sit ele. os Ve 8 yeh 5 bie ke i. ie . een MaMa Wig Cras cay HRA? ee ae x pati ie h.eetry wi Se a ih ride a fhivacntho oume pone ‘st yaa ae Paermatie DAP pskver (AAT) dikyys, Mr eeatag Tatts nhs AMC AE eae : oo ane OA Wea Le eerie cc) sh: $6 et" i ah Mesa eae 7 7 : i ' pe tet ls rel pedal verte the: lesen ar eds) ie epoca) Fiaeit merece MEST Nae wv ata martina: ay alan (6 u wie wwe ne EN We f hit Br TR, bo ail pom an | a ie seh ja Yen O1y ery A we. woh) 1. ; Avoidance of Ultraviolet-B Radiation in Frogs and Tadpoles of the Species Litoria aurea, L. dentata and L. peronii T.F. VAN DE MorTEL! AND W.A. BUTTEMER? 'Faculty of Health Sciences, Southern Cross University, PO Box 157, Lismore NSW 2480; and *Dept. of Biological Sciences, University of Wollongong, Wollongong NSW 2522 VAN DE MoRTEL, T.F. AND BUTTEMER, W.A. (1998) Avoidance of ultraviolet-B radiation in frogs and tadpoles of the species Litoria aurea, L. dentata and L. peronii. Proceedings of the Linnean Society of New South Wales 119, 173-179. Exposure to increasing levels of ultraviolet-B (UV-B) radiation due to anthropogenic reduction of stratospheric ozone (O3) has been suggested as a possible cause of amphibian population declines. Amphibians routinely bask in sunlight as a means of raising their body temperature to levels at which they can assimilate their food more effectively, but unless they are able to detect and behaviourally avoid high-level UV-B radiation, they risk sustaining UV- B-induced DNA damage if basking occurs when O3 levels are low. It is possible that a differ- ential ability to detect and avoid UV-B radiation may be responsible for differences in the population stability of some amphibian species. When placed in an artificial environment which allowed a choice between high-level UV-B radiation versus UV-B-free conditions, lar- val L. aurea and L. peronii were observed more frequently in the UV-B-free environment (p = 0.004 and 0.04), while tadpoles of L. dentata and adult L. aurea and L. peronii showed no significant preference. There were no significant differences between species in the propor- tion of times they were observed under the UV-B-blocking filters. These results suggest that differences in population stability between these species are unlikely to be due to a differen- tial ability to detect and respond to peaks in UV-B radiation. Manuscript received 29 July 1997, accepted for publication 19 November 1997. Keywords: Ultraviolet-B radiation; amphibians; Green and Golden Bell Frog; endangered species; behaviour. INTRODUCTION Exposure to increased ultraviolet-B radiation (UV-BR) due to anthropogenic reduction in stratospheric ozone (O3) has been suggested as a possible cause of wide- spread amphibian population declines (Carey 1994; Grant and Licht 1994; Blaustein et al. 1994; Blaustein et al. 1995; Long et al. 1995; Kiesecker and Blaustein 1995). Due to anthropogenic production of chlorofluorocarbons, stratospheric O3 levels have declined leading to increases of up to 45% (in non-polar areas) in the amount of UV-BR reaching the earth’s surface, mainly in the past two decades, and particularly at montane sites and/or at mid-high latitudes (Atkinson et al. 1989; Blumthaler and Ambach 1990; Roy et al. 1990; Correll et al. 1992; Blumthaler 1993; Frederick et al. 1993; Kerr et al. 1993; Kerr and McElroy 1993). Many amphibian declines have also occurred in relatively undisturbed habitat in montane areas at mid-high latitudes (Corn and Fogelman 1984; Heyer et al. 1988; Richards et al. 1993; Fellers and Drost 1993; Pounds and Crump 1994; Wake 1991) over a similar time frame (Wake and Morowitz 1991). Amphibians are ectotherms (Phillips 1994) and may bask in sunlight as a means of raising their body temperature to levels which optimise physiological functions such as digestion and growth (Wikelski and Trillmich 1994; Christian and Bedford 1995). Proc. LINN. SOc. N.S.W., 119. 1998 174 ULTRAVIOLET-B RADIATION IN LITORIA Ectotherms may have evolved mechanisms to deal with natural levels of UV exposure, however, unless they are capable of detecting and avoiding peaks in UV-BR, they risk UV-B-induced DNA damage if basking occurs when O3 levels are low. As environmen- tal temperature decreases with increasing altitude (Spencer and Grimmond 1994; Vyverman and Sabbe 1995), amphibians at higher altitudes may spend more time bask- ing than lowland populations to reach and maintain their optimal body temperature, thus increasing their exposure to UV-BR. Moore (cited in Philips 1994) suggested that certain amphibian species may detect and avoid UV-BR. Moore exposed three amphibian species to 10 times the normal out- door level of UV-BR in containers that had one area fitted with filters which blocked UV-BR transmission. The African Clawed Frog (Xenopus laevis) and the Pacific Tree frog (Hyla regilla) both of which have stable populations, immediately sought cover behind the filter, whilst the Cascade Frog (Rana cascadae) which is a declining species, basked in the unfiltered light. In Australia, the Green and Golden Bell Frog (L. aurea), which has disappeared completely from high elevation sites (Osborne 1990; White 1995; Osborne et al. 1996) and suffered a population decline at other historic sites, is often seen basking (Barker and Grigg 1977). Sympatric congeners, Litoria dentata and L. peronii, which appear to have stable populations, are not noted for their basking behaviour (pers. obs.). The aim of this study was to determine if larval or adult L. aurea, L. dentata and L. peronii would avoid UV-BR, when given a choice between a UV-B-free and a high UV- B environment. If these species actively avoided exposure to abnormally high doses of UV-BR, one would expect them to spend more time in the UV-B-free environment. If there was a difference between species in the level of avoidance of UV-B wavelengths, one would expect the time spent in the UV-B-free environment would differ between species. This experiment assumed that the frogs would move randomly if they were unable to detect UV-BR. Detection of UV wavelengths may be visual or perhaps via sen- sors which register radiation damage. MATERIALS AND METHODS UV-B Irradiation Irradiation of the frogs and tadpoles was carried out using a cabinet in which one 30 watt and two 40 watt Sankyo Denki UV-B tubes were mounted in brackets on the ceiling. The total number of available watts was limited to 102 by dimmer circuits. The roof of the cabinet was lined with aluminium foil to ensure uniform irradiance. A cellu- lose triacetate filter (supplied by F. Wilkinson CSIRO) was used to remove wavelengths below 290 nm which are not part of the solar spectrum at the earth’s surface. UV-B lev- els were measured using an IL 1350 radiometer/photometer from the University of Sydney. Radiation levels were uniform in the experimental area beneath the lamps. Ultraviolet-B levels between 290 and 300 nm were approximately three times the normal level of UV-BR received at midday on a cloudless summer’s day in the Sydney area. The wavelengths between 290 and 300 nm are both the most biologically active, and those most susceptible to Oz depletion, as they are more strongly absorbed by O3 than the longer wavelengths (Webb 1991). Experimental design Adults This experiment was carried out on L. aurea and L. peronii. Adult frogs used for these experiments were collected from free-living populations in the Illawarra region. Proc. LINN. Soc. N.S.W., 119. 1998 T.F. VAN DE MORTEL AND W.A. BUTTEMER 175 Six adults of each species were placed in separate 30 cm | x 13 cm w x 11.5 cm h plastic boxes in a UV-B cabinet, 18 cm below the UV-B source. The experiments were carried out over two observation periods as the cabinet was not large enough to accommodate 12 animals at one time. Three adults of each species were tested during a given observation period, with each species being placed in alternating boxes. To maintain a balanced design, all replicates used the equivalent number of animals of both species. Half of each box was covered by a Mylar filter which blocked 100% of UV-B wavelengths between 280 and 315 nm, and 90% UV-BR at 320 nm. Mylar transmits 88% (mean value) of visi- ble light (400-700 nm), rising from 85% at 400 nm to 90% by 540 nm (S. Robinson, University of Wollongong pers. comm.). The remaining half of the box was covered by fine polyethylene (Gladwrap) which is transparent to wavelengths in the UV-B range and above. The filters were tested for UV-B (290-320 nm) transmissivity using a Carey 17 spectrophotometer. Ideally, half of the individuals should have been under the UV-B-blocking filter and the other half under the UV-B-transparent filter upon commencement of the experi- ment. However, this was impossible to arrange due to the tendency of the animals to move around after being handled. As a result, the animals were allowed to settle for 15 minutes prior to commencing each experiment. The animals’ position under the filters was noted upon commencement of the experiment and at 15 minute intervals over a peri- od of three hours. The frogs were observed through small holes in a blanket placed in front of the UV-B cabinet to prevent disturbance of the animals by movement of the observer. Air temperature was measured in each container throughout the experiment using thermocouples. Tadpoles Tadpoles were reared from spawn collected for the experiments reported by van de Mortel and Buttemer (1996). The experiment was carried out on L. aurea, L. dentata and L. peronii. Ten tadpoles of a given species were placed in each 30 cm 1 x 13 cm w x 11.5 cm h plastic container in 2 cm of water and placed in the UV-B cabinet. There were eight replicates per species. The species were placed in alternating containers, which were covered with filters as above. At time zero, 50% of the tadpoles were under the UV-B-blocking filter. The tad- poles’ position under the filters was noted at 15 minute intervals over a period of 1.5 hours. The tadpoles were observed through small holes in a blanket placed in front of the UV-B cabinet, to prevent disturbance of the animals by movement of the observer. Water temperature was measured in each container during the experiment as described above. Statistical analysis The number of animals counted under the UV-B-blocking filter was converted into a proportion of all observations for each replicate. An arcsine of the square root transfor- mation was used to improve the normality of the distribution (Zar 1984). The effect of species on the choice of environment was tested using a single factor ANOVA for the data obtained from the tadpole experiment, and using a t-test for the data obtained from the experiment on adults. Sign tests were carried out for each species to determine if the proportion of counts under the UV-B-blocking filters was greater than those under the UV-B-transparent filters (Zar 1984). Sign tests were used to avoid conflicts with the independence of observations within each replicate. As Sign tests have a low power to reject a false null hypothesis, power analyses were carried out on those data which did not show a significant difference (Zar 1984). All results are reported as mean values + the standard error unless noted otherwise. Proc. LINN. SOc. N.S.W., 119. 1998 176 ULTRAVIOLET-B RADIATION IN LITORIA RESULTS Adults Litoria aurea and L. peronii adults were observed under the UV-B-blocking filters on 71 + 17% and 83 + 17% of occasions, respectively (n=12) (Fig. 1). Counts under the UV-B-blocking filters were not significantly different to those under the UV-B-transpar- ent filters for either L. aurea (p=0.11) or L. peronii (p=0.09). The power of those analy- ses was 0.09 and 0.33, respectively, which indicates a very low probability of correctly rejecting a false null hypothesis. There were no significant differences between species in time observed under the UV-B-blocking filters (t=-0.481; df=10; p=0.64). Fifty per- cent of both L. aurea and L. peronii did not move from the location they held at time zero. There was no trend of greater movement to the UV-B-free environment as time spent under the UV-B source increased. Air temperature in each replicate was 27.8-27.9°C. Tadpoles Litoria aurea (p = 0.004) and L. peronii (p = 0.04) were observed on a significantly greater number of occasions under the UV-B-blocking filters than under the UV-B-trans- parent filters, while L. dentata was not (p=0.06). The power to reject a false null hypothe- sis for the L. dentata data was 0.35. Litoria aurea, L. dentata and L. peronii tadpoles were observed under the UV-B-blocking filters on 74 + 3%, 68 + 6% and 67 + 4% of occasions, respectively (Fig. 2). There were no significant differences between species in time spent under the UV-B-blocking filters (F = 0.69; df = 2, p = 0.51). Water temperature of the replicates ranged from 25.4-25.7°C. There was a slight non-significant trend of move- ment to the UV-B-blocking filter as time of exposure to UV-BR increased (Fig. 3). Mean proportion of counts under UV-B-blocking filter L. aurea L. peronii Species Figure 1. The mean (+ sem) proportion of all observations in which Litoria aurea and L. peronii adults were situated beneath the UV-B-blocking filter. Six adults of each species were exposed to three times the normal midday summer level of UV-BR at wavelengths between 290-300 nm. Proc. LINN. SOC. N.S.W., 119. 1998 T.F. VAN DE MORTEL AND W.A. BUTTEMER 177 Mean proportion of counts under UV-B-blocking filter L. aurea L. dentata L. peronii Species Figure 2. The mean (+ sem) proportion of all observations in which Litoria aurea, L. dentata and L. peronii tadpoles were situated beneath the UV-B-blocking filter. Eighty tadpoles of each species were exposed to three times the normal midday summer level of UV-BR at wavelengths between 290-300 nm. 1.0 0.9 0.8 0.7 0.6 filters 0 15 30 45 60 75 90 Proportion of tadpoles under UV-B-blocking Time (minutes) Figure 3. Proportion of Litoria aurea, L. dentata and L. peronii tadpoles under the UV-B-blocking filters as a function of time. Eighty tadpoles of each species were exposed to three times the normal midday summer level of UV-BR at wavelengths between 290-300 nm. Ml = L. aurea, A = L. dentata, O = L. peronii. Proc. LINN. SOc. N.S.W., 119. 1998 178 ULTRAVIOLET-B RADIATION IN LITORIA DISCUSSION There were no significant differences in the number of occasions that adult L. aurea and L. peronii, and L. dentata larvae were observed under the UV-B-blocking fil- ters, however, the power of these experiments to reject a false null hypothesis was very low. With a sample size of six animals per species, 100% of the animals needed to be observed more frequently under the UV-B-blocking filter than under the UV-B-transpar- ent filter to show a significant difference. In contrast to the adults, larval Litoria aurea and L. peronii were observed significantly more often under the UV-B-blocking filters than under the UV-B-transmitting filters. These results suggest that tadpoles of these species may detect and avoid high doses of UV-BR. It is possible that the tadpoles were responding to a decrease in visible wavelengths under the Mylar (UV-B blocking filter), although the 12% average reduction in visible wavelengths was not visible to the observers. Unfortunately, it is not possible to block UV-B wavelengths without affecting visible wavelengths as well. There was no thermal gradient between the two experimental treatments as recorded temperatures were the same under both filters. The results of this experiment differed from those of Moore (reported in Phillips 1994), who found that the declining species (R. cascadae) did not avoid UV-BR, while those species with apparently stable populations did. It is difficult to compare Moore’s results to those of the present study because little information is available regarding details of his experimental design. However, the level of exposure in Moore’s experiment seemed excessive (10 x the normal level of UV-B radiation) as the greatest demonstrated increase in UV-BR in areas with amphibian populations has been a short term increase of 45% (Frederick et al. 1993). It may be that testing L. aurea, L. dentata and L. peronii under considerably higher levels of UV-BR would have elicited a significant difference between species, however, results obtained under these conditions could not realistically be extrapolated to species in the field. Further work with a larger sample size is needed to clarify our results, particularly as the more powerful parametric tests used to analyze the effect of species on filter choice showed no significant differences between species in the number of occasions the animals were found under the UV-B-blocking filters. These results suggest that differ- ences in population stability between these species are unlikely to be due to a differential ability to detect peaks in UV-BR. ACKNOWLEDGEMENTS We are grateful to Gavin Greenoak for his assistance, comments and ideas. Permission to collect ani- mals for this experiment was granted by the National Parks and Wildlife Service, Permit B1292. REFERENCES Atkinson, R., Matthews, W., Newman, P. and Plumb, R. (1989). Evidence of the mid-latitude impact of Antarctic ozone depletion. Nature 340, 290-293. Barker, J. and Grigg, G. (1977). ‘A field guide to Australian frogs’. (Rigby: Adelaide). Blaustein, A., Hoffman, P., Hokit, D., Kiesecker, J., Walls, S. and Hays, J. (1994). UV repair and resistance to solar UV-B in amphibian eggs: a link to population declines? Proceedings of the National Academy Sciences, USA 91, 1791-1795. Blaustein, A., Edmond, B., Kiesecker, J., Beatty, J. and Hokit, G. (1995). Ambient ultraviolet radiation causes mortality in salamander eggs. Ecological Applications 5,740-743. Blumthaler, M. and Ambach, W. (1990). Indication of increasing solar ultraviolet-B radiation flux in alpine regions. Science 248, 206-208. Blumthaler, M. (1993). Solar UV measurements. In ‘UV-B radiation and ozone depletion: effects on humans, animals, plants, microorganisms and materials’ (Ed. M. Tevini) (Lewis Publications: Boca Raton). Proc. LINN. SOC. N.S.W., 119. 1998 T.F. VAN DE MORTEL AND W.A. BUTTEMER 179 Carey, C. (1994). Role of stress in amphibian declines and extinctions. Second World Congress of Herpetology: Abstracts p. 48-49. Christian, K. and Bedford, G. (1995). Seasonal changes in thermoregulation by the frillneck lizard, Chlamydosaurus kingii, in tropical Australia. Ecology 76, 124-132. Corn, P. and Fogelman, J. (1984). Extinction of montane populations of the northern leopard frog (Rana pipiens) in Colorado. Journal of Herpetology 18, 147-152. Correll, D., Clark, C., Goldberg, B., Goodrich, V., Hayes, D., Klein, W. and Schecher, W. (1992). Spectral ultraviolet-B radiation fluxes at the earth’s surface: long-term variations at 39N, 77W. Journal of Geophysical Research 97, 7579-7591. Fellers, G. and Drost, C. (1993). Disappearance of the Cascades Frog Rana cascadae at the southern end of its range, California, U.S.A. Biological Conservation 65, 177-181. Frederick, J., Soulen, P., Diaz, S., Smolskaia, I., Booth, C., Lucas, T. and Neuschuler, D. (1993). Solar ultravio- let irradiance observed from Southern Argentina: September 1990 to March 1991. Journal of Geophysical Research 98, 8891-8897. Grant, K. and Licht, L. (1994). Effects of ultraviolet on life history parameters of frogs from Ontario, Canada. Second World Congress of Herpetology: Abstracts p. 101. Heyer, W., Rand, A., da Cruz, C. and Peixoto, O. (1988). Decimations, extinctions and colonizations of frog populations in Southeast Brazil and their evolutionary implications. Biotropica 20, 230-235. Kerr, J., Wardle, D. and Tarasick, D. (1993). Record low ozone values over Canada in early 1993. Geophysical Research Letters 20, 1979-1982. Kerr, J. and McElroy, C. (1993). Evidence for large upward trends of ultraviolet-B radiation linked to ozone depletion. Science 262, 1032-1034. Kiesecker, J.M. and Blaustein, A-.R. (1995). Synergism between UV-B radiation and a pathogen magnifies amphibian embryo mortality in nature. Proceedings of the National Academy of Science, USA 92, 11049-11052. Long, L., Saylor, L. and Soule, M. (1995). A pH/UV-B synergism in amphibians. Conservation Biology 9, 1301-1303. Osborne, W.S. (1990). Declining frog populations and extinctions in the Canberra region. Bogong 11, 4—7. Osborne, W.S., Litttlejohn, M.J. and Thomson, S.A. (1996). Former distribution and apparent disappearance of the Litoria aurea complex from the Southern Tablelands of New South Wales and the Australian Capital Territory. Australian Zoologist 30, 190-198. Phillips, K. (1994) ‘Tracking the vanishing frogs: an ecological mystery’. (Penguin: New York). Pounds, J. and Crump, M. (1994). Amphibian declines and climate disturbance: the case of the Golden Toad and the Harlequin Frog. Conservation Biology 8, 72-85. Richards, S., McDonald, K. and Alford, R. (1993). Declines in populations of Australia’s endemic tropical rain- forest frogs. Pacific Conservation Biology 1, 66-77. Roy, C., Gies, H. and Elliott, G. (1990). Ozone depletion. Nature 347, 235-236. Spencer, N. and Grimmond, N. (1994). Influence of elevation on the thermoregulation of two sympatric lizards. N.Z. Journal of Zoology 21, 379-385. van de Mortel, T. F. and Buttemer, W.A. (1996). Are Litoria aurea eggs more sensitive to ultraviolet-B radia- tion than eggs of sympatric L. peronii or L. dentata? Australian Zoologist 30, 150-157. Vyverman, W. and Sabbe, K. (1995). Diatom-temperature transfer functions based on the altitudinal zonation of diatom assemblages in Papua New Guinea — a possible tool in the reconstruction of regional palaeocli- matic changes. Journal of Paleolimnology 13, 65-77. Wake, D. (1991). Declining amphibian populations. Science 253, 860. Wake, D. and Morowitz, H. (1991). Declining amphibian populations — a global phenomenon? Findings and recommendations. Alytes 9, 33-42. Webb, A. (1991). Solar ultraviolet radiation in southeast England: the case for spectral measurements. Photochemistry and Photobiology 54, 789-794. White, A. (1995). Green and Golden Bell Frogs. Frogfacts 5, 1-4. Wikelski, M. and Trillmich, F. (1994). 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N.S.W., 119. 1998 ea ante nyu, rida iil reat vee ask ER ) Ce et is PBs sor a sn wai 0 ect aki ae i Rowsttrse ol ives = om et oi = : t apc i: =: ee eee RSE ee . 2 vil. aot Siar ie sided vmsaik eiheae eae \ineunees fe yu ac - hasonansy spcdib Sait RAIN TIT ESS, ake Bh.” 72) Pitotanys ye id : SNe ett FOUN [iat Shes sot | a Z ra , ws Atal halt Hhufivke 2 ath nh n iene Vie: 1K Homey iG tir. we eS *é fa ae ei] p= be oT we a Weneaey as ; : Cie t9 FE cals y A (Se £45 soar, enelindi|n a5 as eleai gi at moses Gn se Ling Rgnibatl Taosaionodg Laing 3p is pA CAE TARID Blah jBOL) 4 aiden OM Brut a == = = * 7? - eS Hib tel ceee Ee, LB age i I or) ert) a} 8a ees 3 cm An anh) re War Street nogeqatnn, si Es) 29Epsede PTst hy chr i OTL 2% RO iva moras oxin Oo onaaine pop sania iP povertpe om G . 3 : jt fnotges: rAlreegnac ite “elec: tamale \- c Te ‘ pee steay &* mi Vigra i [2 dae) Sa Ra, A si Paik “ ie fue se ie _ A of ‘ ‘ ‘io ~ ee r =i lea His ; fi 3 . ’ 1avite act - oS) eubAr Ww AY yids fu: _ ‘iReiatiy ‘V, et eran 7 7 Wier, me. oy a ii al Cee 2 ack lays ] fete SF y Vv epee So iL427 1? GjeM2 “op m, da} ih aha er a PProrenaitys a ihe Masked ’ or i ‘ef, >> ih ejenaed © Bali yt. Berwiny ¢. Beats Sait Hokit, yy, (nib Asubt utes, ate ton tae? > i oo oe S;yliege Mone’ ‘Sah ee an = o ani die a be "a Wath pri oo mere be sien i i ait ey, uy , ae See 6, SR - . Gieahaniot? (7505)-Ac'o 1? aoe, 4 “0 NB Stents ‘i dendarea). PAU, thsi tyattago gallon carer’ 404 a" Tou! it iaw AE Glow, yooh? a sie Proglacial Hydrology and Drainage, Southeastern Vestfold Hills, East Antarctica DAMIAN B. GORE! AND JOHN PICKARD? 'School of Earth Sciences, Macquarie University, NSW 2109. dgore @laurel.ocs.mq.edu.au; and *Graduate School of Environment, Macquarie University, NSW 2109. john.pickard @ gse.mq.edu.au Gore, D.B. and Pickard, J. (1998). Proglacial hydrology and drainage, southeastern Vestfold Hills, East Antarctica. Proceedings of the Linnean Society of New South Wales 119, 181-196. Three ice dams in southeastern Vestfold Hills, East Antarctica, dam a system of five lakes periodically, impounding more than ~1.5 x 10° m’ of water. Dam #a impounds 1.1 x 10° m* of water, while Dams #b and #c prevent the free drainage of the lake below Dam #a, and impound the remaining 0.4 x 10° m’. The mode of failure of these dams and the rate of impoundment release were not known until January 1993, when Dams #a and #b failed, allow- ing a flood to travel along a channel incised in sediment, and into Crooked Lake at >8 m’s'; four times the peak midsummer discharge of the largest stream in Vestfold Hills. The flowpath from Lake #10 is determined by which of two dams fails first; the northwestern dam (#b) allows the impoundment to travel into Crooked Lake via Grimmia Gorge (observed during January 1993), and the northern dam (#c) into Crooked Lake via Sickle Lake, Lake Verkhneye and Foot Lake (observed during 1979 and 1990). Formation and failure of these Vestfold Hills ice dams is similar to “snow dams’ described from the Canadian Arctic. Floods released from the failure of the Vestfold dams provides an alternative explanation for a sudden increase in dis- charge at Ellis Rapids in January, 1976. Descriptions in this paper of glaciofluvial features are at odds with published notions that such features are absent from Vestfold Hills. Manuscript received 20 August 1997, accepted for publication 10 December 1997. KEYWORDS: Channel reach, fluvial, flood, hydrology, ice dam, melt, sublimation. INTRODUCTION Vestfold Hills is a 413 km’ area in East Antarctica (Fig. 1) that contains several externally draining freshwater stream systems (Tierney 1975; Colbeck 1977; Adamson and Pickard 1986; Bronge 1989; Gore 1991, 1992). Some workers (Hirvas and Nenonen 1991; Hirvas et al. 1993, 1994a, 1994b) argue that an apparent lack of glaciofluvial fea- tures in Vestfold Hills is evidence for deglaciation by sublimation and not melt. Glaciofluvial features do exist however: Gore (1991, 1992) described the upper reach of a large channel cut by fluvial activity into sediment in southeastern Vestfold Hills, but did not visit or describe the lower reaches. The first aim of this paper is to complete the description of that drainage system, to add to the growing body of evidence on glacioflu- vial features in Vestfold Hills. Gore (1991, 1992) speculated that the upper reach of the channel had been incised by flows released from a failed ice dam. During 1993 we were fortunate to be camped beside the channel when a flood occurred. The second aim of this paper is to describe this 1993 flood and its origin at the system of ice dams upstream, and refer to the drainage model hypothesised by Gore. Floods released from these ice dams contribute flow to the largest drainage net- work in Vestfold Hills, the Druzhby system (Fig. 1), which has a peak measured dis- charge at the outlet at Ellis Rapids of 2 m’s' (Colbeck 1977). Bronge (1989) selected a Proc. LINN. SOc. N.S.W., 119. 1998 182 PROGLACIAL HYDROLOGY IN EAST ANTARCTICA Tierney _4 River B oa Key lake & stream with flow direction es wy _, boundary of Creek” * derennial ice & snow 29. ix.80 date runoff first observed (1980/81 season) Figure 1]. Vestfold Hills showing location of the Druzhby drainage system, with places mentioned in the text. The main area of interest in southeastern Vestfold Hills, is outlined. The date of onset of runoff during the 1980/81 summer is shown. Proc. LINN. SOC. N.S.W., 119. 1998 D.B. GORE AND J. PICKARD 183 sub-catchment of the Druzhby system at Tierney Creek (Fig. 1), and recorded water depth, short-wave radiation, air temperature and water temperature over a 13 week peri- od in the 1987/88 summer. Bronge found that during the earlier part of the melt season, discharge was best correlated with irradiance, while in the latter part of the melt season, discharge was best related to air temperature. After incorporation of a time lag factor to allow water to reach the outlet from distant parts of the catchment, Bronge demonstrated that discharge can be related closely to these meteorological variables. However, Colbeck (1977) recorded a discharge pattern that was partly unrelated to these meteoro- logical parameters. Colbeck observed a sudden increase in discharge at Ellis Rapids between 15-20 January 1976. He hypothesised that Crooked Lake upstream had fallen below its outlet sill due to winter ablation, filled during the first part of the melt season and finally started to overflow in mid-January. The hypothesis of Crooked Lake having fallen below its outlet sill during late winter to early spring is also supported by our observations at the start of the melt season. However, it is possible that the release of reservoirs ponded behind ice dams could create short-lived, but dramatic increases in the discharge hydrograph downstream, that are unrelated to meteorological variables. The third aim of this paper is to present this alternative explanation for the discharge pattern at Ellis Rapids documented by Colbeck (1977). Ponding, diversion and sudden release of water by barriers of ice and snow in the present day proglacial zone creates drainage patterns and discharge rates that are rather different to those that might occur should the area become free of ice. The final aim of this paper is to contend that accurate reconstructions of the complexity of former proglacial flowpaths in low-gradient, deglaciated areas, using the fragmentary distribu- tion of fluvial features, are rather difficult, if not impossible. RESULTS Hydrology and the formation of ice dams from wind-blown snow In this paper, we define the flowpaths of southeastern Vestfold Hills as those depicted on Fig. 2. Bronge (1989) identified the most important meteorological variable for early summer melt generation as irradiance, and late summer melt generation as air temperature. Our field observations of melt generation support these findings. On the ice sheet early in the melt season, at air temperatures as low as —10°C, it is possible to observe ice crystals in full sunlight, detaching and falling from ice cliffs. On more gentle slopes, ice crystals detach but remain in situ until enough water is generated to make them flow as an ice-laden slurry. Movement of these slurries causes ice crystals at their margins to become stranded, leaving levees that are reminiscent of some terrestrial debris flows. As the air warms toward 0°C, and incoming solar radiation remains undiminished by cloud, melt streams form on the ice sheet and flow toward the land. The first flows that reach the land are absorbed into snowpack that accumulated during previous winters. In this way, there is a significant lag between the onset of runoff near to the ice edge and at the outlet of the Druzhby system. Snowbanks that lie in the lee-side of hills pond runoff into pools of variable size and longevity. Snow by definition is permeable, but the interiors of these snowbanks have not yet warmed to ambient summer temperatures, and flows that soak into them typically freeze solid for some time, allowing ponding of water behind the snow/ice dam. The duration of ponding is dependent on several factors - among them size and density of the snowbank dam, the extent of ice within the dam, and the temperature and volume of water. In most cases, small snow/ice dams last from a few hours to a few days at most (e.g. Woo and Sauriol 1980; Pickard 1986:297), but where the snow is thick and dense, the ice that forms within the snowpack may last for several years. The internal structure within the ice dams in southeastern Vestfold Hills can be Proc. LINN. Soc. N.S.W., 119. 1998 184 PROGLACIAL HYDROLOGY IN EAST ANTARCTICA |Crooked iLake lake & stream with flow direction m= ice dam channel reach _ perennial ice & snow, ge Fe Figure 2. Drainage and ice dams in southeastern Vestfold Hills. Many smaller ice and snowbank dams are not shown here, for clarity. UR=upper reach, MR=middle reach, LR=lower reach, FL=Foot Lake, LV=Lake Verkhneye, SL=Sickle Lake, BH=Boulder Hill. Lakes are numbered 1—13; ice dams are coded a-f. seen in tension cracks and outlet channels: the ice is layered, less dense than glacier ice and glacial debris is absent. These observations are strong evidence that the ice dams have formed from the in situ transformation of accumulated wind-blown snow into ice, as a result of saturation by summer meltwater with subsequent deep freezing during win- ter, as well as compaction. The ice-dammed lakes A group of four lakes in the southeast corner of the Vestfold Hills (Lakes 1-4, Fig. 2), are dammed periodically by a barrier made of ice and wind-drifted snow (Ice Dam #a). The lakes rise as much as five metres above their outlet sill heights, impounding an additional c. 1.1 x 10° m* of water (Gore 1992), and partially drain when the dam fails. As the annual addition of wind-drifted snow, saturated with meltwater, refreezes at the end of summer, the dam reforms into an impermeable ice barrier. Sequential aerial pho- tographs and satellite imagery from 1947 to 1990, and fieldwork during the 1979/80, 1988/89, 1989/90 and 1992/93 summers revealed that Ice Dam #a has been in existence from at least 1947, and that failure of that dam occurred during the 1987/88, 1989/90, 1991/92 and 1992/93 summers. We speculate that many more failures have occurred, but due to infrequent visitation they have never before been observed. The occurrence of ice-damming in this area first came to light with two discover- ies. The first was that Lakes 1-4, in the January 1987 aerial photographs, were flooded and joined to form one large lake. This highlighted the location of Ice Dam #a (Fig. 2). The second was the discovery of a dry channel which trends west-southwest from a large snowfield, around the south side of Boulder Hill and through Grimmia Gorge into Proc. LINN. SOc. N.S.W., 119. 1998 D.B. GORE AND J. PICKARD 185 Crooked Lake (Fig. 2). Ice Dam #a had never been observed to fail, and due to large, >10 m deep snow and ice fields draping the underlying topography below Lake 10 (Fig. 2), it was not known with certainty whether the channel was related to the ice dam or not. Gore (1992) hypothesised that the failure(s) of Ice Dam #a released flows which created the channel. Lake 10, below Lakes 1-4 and Ice Dam #a, is partially impounded by two ice dams, #b to the northwest and #c to the north (Fig. 2). Observations of silt lines on rocks around the lake, and lake height during the 1993 draining show that the lake may fall more than 5 m following failure of the ice dam, releasing more than 0.4 x 10° m’ of water. The exact dimensions of the three ice dams are uncertain, since they merge into surrounding snow and ice fields. However, Ice Dam #a is approximately 100 m wide, 200 m long and perhaps 20 m deep, #b is 30 m wide, 100 m long and 5 m deep, and #c is approximately 100 m wide, and at least 20 m deep (the length was not able to be deter- mined). Ice Dams #a and #c are part of broad snow- and ice-filled depressions, whereas Ice Dam #b is a lee-side snow and ice accumulation behind a small bedrock hill. Several other ice dams exist in southeastern Vestfold Hills. Ice Dam #d impounds the small Lake 11, that has a volume of 0.061 x 10° m’, while Dam #f impounds the much larger Lake 13; however no dam thickness or water depth data are available. Streams with midsummer discharges up to 0.5 m’s' flow into Lake 13 from the ice sheet, through Dam #e. While no outflow has been observed from Lake 13, bedrock barriers to the north and south allow us to infer that water flows periodically into Lake Verkhneye (LV on Fig. 2) to the southwest when Ice Dam #f fails. The three channel reaches The three channel reaches marked in Fig. 2 are the largest fluvially incised flow- paths in sediment yet found outside of the Transantarctic Mountains (e.g. Selby 1971; Chinn and Dickson 1986). With a maximum width of 30 m and depth of 4 m, the 500 m long upper reach was formed by the removal of more than 30,000 m* of glacial valley fill (Gore 1992). In places, fluvially derived fine gravel deposits mantle the right bank (Fig. 3), particularly where the channel widens, palaeofloods became shallower and lost com- petence and dropped the coarsest fraction of their sediment load. Surveyed cross-sections (T1 and T2 on Fig. 3) revealed that the left bank scarp is steeper than the right bank, reflecting undercutting during floods. Observations during 1993 of places where the glacial sediment forming the left bank had been undercut and was collapsing onto the channel floor, is strong evidence for recent flood(s) along the channel. The incision of the upper reach is controlled by a rock bar at the downstream end. Some 300 m of bedrock channel lie between the upper reach and Lake 12 (Fig. 2). A further 200 m of boulder-strewn rock channel lie between Lake 12 and the middle reach. The middle reach consists of incised glacial valley fill. Figure 4 shows a clearly demarcated flood trimline in the confined, bedrock section of the flowpath where palae- ofloods up to ~8 m above the present valley floor have removed glacial sediments. Some of those sediments were deposited in bars (?) on the right bank immediately downstream where the channel is wider and where hydraulic competence of former floods must have been less than in the more confined parts of the flowpath. The depositional setting is analogous to the bars on the upstream end of upper reach (Fig. 3). In places on the left bank of middle reach, erosional scarps reveal matrix-supported, silty glacial sediment, exposed by the palaeofloods that have scoured the channel from time to time. The clearly defined part of the stream channel varies from <20 m width at the upstream end to >50 m width at the head of Grimmia Gorge, and the height of washed debris lies in the range 6—8 m above the thalweg (Figs 4 and 5). The fluvially modified parts of T3—T6 (Fig. 5), with a mean cross-sectional area of 370 m’, allow estimation of the volume of glacial debris removed to form the middle reach as ~130,000 m°. Terraces Proc. LINN. Soc. N.S.W., 119. 1998 186 PROGLACIAL HYDROLOGY IN EAST ANTARCTICA Figure 3. Oblique aerial photograph of the upper reach of the channel, looking downstream. Tl and T2 mark the locations of transects described and discussed by Gore (1991, 1992). Note the fine gravel mantling the right bank at lower right, and the outline of the right bank scarp produced by the shadow. Fluvial deposits on the lee- side of boulders may be found across all of the valley fill. Ice lines the edges of the stream, which is shown here with a discharge of <2 m’s'. The feature named ‘delta?’ may be an erosional remnant of the valley fill, rather than a delta which is strictly depositional. Lake 11 drained into the unnamed pond at top left within the previous few months (see text). The channel at T2 is 40 m wide. Photo taken 29 January 1993. lie on the left and right bank immediately above Grimmia Gorge (Figs 4 and 5). The upper limit of the terraces on both sides of the channel coincide, allowing us to conclude that they were created by fluvial processes. However, it was not possible to determine whether these terraces, or indeed any of the washed sediments, were created during con- temporary floods, were stranded following downcutting of the valley fill, or were formed as shorelines during times of an ancestral Glacial Crooked Lake (Gore et al. 1996). The 350 m long middle reach ends with a rock bar immediately above Grimmia Gorge; a flu- vially cut slot gorge ~80 m long, with a cliff at its head of some 10 m height. The lower reach is a fluvially moulded and scarped, and partly dissected boulder- strewn deposit (Fig. 6). It has the morphology of a fan/delta, that has been formed and Proc. LINN. SOc. N.S.W., 119. 1998 D.B. GORE AND J. PICKARD 187 Crooked Lake Figure 4. Oblique aerial photograph montage of the 350-m long middle reach of the channel. Flow is from right to left; at Grimmia Gorge the flow is away from the viewer. The flood-scoured bedrock channel is identified at right, as are the bars immediately downstream on the right bank. The four surveyed transects are located. The lower reach is here coded f/d 1, for fan/delta 1. Photo taken 29 January 1993. been incised by multiple floods. Whether or not this feature is a true fan/delta is not known with certainty. As an order of magnitude estimate, if all of the debris removed from the upper and middle reaches upstream were to be distributed across the fan/delta surface, that layer would be some 3 m thick. Since the fan/delta is clearly much thicker than that, additional sources of debris (from previous glacial advances?) with attendant sediment transport mechanisms (floods released from failed ice dams?) are implied. Other fans are found in the same area; f/d 2 is a large depositional feature in the valley west of Grimmia Gorge. Like f/d 1, the boulders on the surface of f/d 2 exhibit a stream- lined pattern that was probably formed from fluvial activity. Another fan/delta lies at the Proc. LINN. SOc. N.S.W., 119. 1998 188 PROGLACIAL HYDROLOGY IN EAST ANTARCTICA 20 18 upper limit of washed 16 <4— debris ie right 12 bank ® 210 (e) 2 9 © 1 20-5 40 60 100 £ 65 16 undercut scarp - © 14 no washed debris 2 12 | 74 iS right £10 bank ® 8 = 20 40 60 80 ; = Le minor : channel pe terrace = © 8] upper limit 7S ae. « ie of washed iar « 6] debris? right S bank 3 4 eo) POCO CON 100m a120n ani 4OsmmmeO oO @ 10 = 8 OP Minar st a terraces —— = 6 upper limit right 4| of washed eral debris 2 0 0 20 40 60 80 100 120 140 160 metres along transect Figure 5. Surveyed transects across the middle reach (looking downstream; location on Fig. 4). The upper limit of washed debris is shown for each transect, where able to be determined. Proc. LINN. SOC. N.S.W., 119. 1998 D.B. GORE AND J. PICKARD 189 Figure 6. Oblique aerial photograph of the area around the lower reach, looking east. The head of Grimmia Gorge is marked gorge, and the lower reach is marked f/d 1. Flow through the gorge is from right to left. The fan/delta at the outlet of an adjacent valley is marked f/d 2. A third fan/delta is visible to the left of the name ‘Foot L’. The end of Foot Lake closest to the viewer is 200 m across. Photo taken 29 January 1993. northeastern corner of Foot Lake (visible at the top left of “Foot L’ on Fig. 6). These fea- tures may be used to argue that palaeofloods large enough to transport boulders, have occurred in a number of other proglacial valley systems. The January 1993 discovery of Ice Dam #d and Lake 11 Sometime during late 1992, Ice Dam #d failed, releasing 0.061 x 10° m’ of water into the upper reach of the channel. During previous summers of work in that area, we had traversed repeatedly across the snow-covered frozen surface of the reservoir, without suspecting its existence. Upon arrival at Ice Dam #d in January 1993, we found a hole in the ice field where the reservoir used to be, and a ‘keyhole’ slot through the ice dam (Fig. 7). We surveyed the hole to determine the volume of the reservoir, and examined the keyhole and tunnel through the ice dam. The keyhole form of the conduit, and subtle notching of the snow and ice by thermal erosion by water (Fig. 7), allow the mechanism by which the ice dam failed to be inferred. Field observations showed that the upper 1.5 metres of the ice dam consisted of permeable firn and snow, while at greater depths the dam consisted of solid ice. The reservoir, upon reaching the ice/firn boundary, seeped out slowly, creating a broad (1.2 m) but shallow (~0.4 m) tunnel which later formed the broad top of the keyhole. As this seepage reached the downstream side of the dam, the discharge rate increased as the tunnel enlarged and was eroded free of snow and firn. Hydraulically efficient pipe flow on the ice surface was able to cut rapidly a vertical tun- nel through the ice dam via mechanical and thermal erosion in a positive feedback - the deeper the tunnel became, the greater the discharge rate, and thus the greater the rate of incision. Without the buttressing afforded by the water, cracks appeared in the walls of Proc. LINN. Soc. N.S.W., 119. 1998 190 PROGLACIAL HYDROLOGY IN EAST ANTARCTICA Figure 7. The keyhole slot through Ice Dam #d which formerly ponded Lake 11. Only the upper 2 m of the dam is visible; the tripod stands at the edge of a 6 m drop to the deepest part of the floor of the drained lake. The reservoir drained through the ice dam some time during late 1992. The broad upper part of the keyhole coincides with a transition from permeable firn to impermeable ice; the tunnel through the ice dam revealed layered ice formed by years of snow accumulation in the lee-side of a hill. Photo taken 22 January 1993. the ice dam (e.g. right side of Fig. 7) as ice slumped into the hole where the reservoir used to be. Ice Dam #d is less than 10 m in thickness (Fig. 8). It surprised us that in six seasons of working in this part of Vestfold Hills we had seen the lake in a drained state only once. This observation suggests that quite thin dams, formed from the in situ trans- formation of wind-drifted snow into ice, can be rather stable. The 25 January 1993 failure of Ice Dam #b Flow discharge, and stage of Lake 12 Early in the morning of 25 Jan 1993 (approximately 0200 h), the inlet to Lake 12 just below the upper reach (Fig. 2), increased in discharge dramatically, as indicated by the noise of the rapids. The first definite evidence of a major flood came at 0930 h, when the inlet of Lake 12 was noticed to have increased in discharge visibly, and Lake 12 had expanded toward the foot of our tent camp. No fieldwork was carried out on the flowpath until the afternoon of the 25th, at which time the discharge was first gauged and the height of Lake 12 had increased measurably (Fig. 9). Stream discharge was estimated using the velocity/area method, with current velocity measured by Pygmy Ott current meter where possible, or by the rate of passage of cut blocks of ice over 30 m where the Ott could not be used safely. The peak velocity of the stream reached at least 1.3 ms" at the surface, with a maximum discharge of >8 m’s'' (Figs 9 and 10). Throughout the ris- ing stage of the flood, Lake 12 acted as a reservoir, delaying and attenuating the passage of the flood. While the data forming Fig. 9 are few, it is possible to estimate the order of Proc. LINN. SOc. N.S.W., 119. 1998 D.B. GORE AND J. PICKARD 19] Key © glacial debris A ice scree #. frozen lake surface + bedrock 7.2 @ spot height (m above lake) 9.8 — -@ — — interfluve between lakes 9.3 Figure &. Plan map of Lake 11 and Ice Dam #d. The upper limit of silt at top right records the former extent and height of the lake in that direction. magnitude of the flood as 1.2 x 10° m*, by calculating the area under the discharge curve. This volume is similar to that impounded by Ice Dams #a-c upstream. Dam location and mode of failure The flowpath was traced along the upper reach to where the flow was issuing from a breached ice dam (Ice Dam #b, Fig. 2) in Lake 10. The location of Ice Dam #b on Fig. 2 is slightly different to that suggested by Gore (1992); instead, Ice Dams #b and #c are separat- ed by only 50 m at the northern side of Lake 10. As at 2200 h on the 25th January, water drained through a steep (~5 m fall over 80 m), smooth ice-bedded channel through Ice Dam #b. The low friction of the ice allowed water to exit the channel at approximately 4 ms‘. It is surprising that the ice floor of the channel was able to withstand flows of this discharge and velocity for more than a few hours, but at that time the flood had been in progress for more than 20 hours. We suspect that Ice Dam #b failed by overtopping and progressive incision, but we have no further observations to support this notion. Aerial photography and field inspection suggests that the discharge rate of Lake 10 may be ultimately controlled by a rock bar buried within Ice Dam #b, although this is not known with certainty. DISCUSSION From the ice sheet to Crooked Lake, the drainage system of southeastern Vestfold Hills can be seen to be rather dynamic, with floods being routed along different drainage routes depending on which of several ice dams fail first. The hypothesis of Proc. LINN. Soc. N.S.W., 119. 1998 192 PROGLACIAL HYDROLOGY IN EAST ANTARCTICA 0.6 is ;! @=———® stream discharge | 0.5 bs (m3s-*) K 2 aa Se -x height of Lake 12 E 8 (m above arbitrary | 0.4 © = Ry! datum) = ) l Q (ap) I © eB ms \ 3-1 r ae Qmax = 8 ms 0.3 @ coe 6 m3 g 5 tN Qtot = 1.2 x 108 m : & a¢ : 02 = © I X ol 2 I = O F g I ==] 1S (©)) 7 ie : o 0K 3 0 = OnsyOlg 5O) OS) oO Si OnkLOonnO© Ska Usenet Ql Mec@ig 1Olza 1 ails shale Ol St mH o =) Nz (2) - 25 Jan 26Jan 27 Jan 28 Jan 29 Jan 1993 Figure 9. Stream discharge and the height of Lake 12 over the five days following the failure of Ice Dam #b. alternative drainage routes posed by Gore (1992) has been substantiated by field evi- dence from 1993, although in the present paper we locate more accurately Ice Dam #b. Large channels cut into sediment and flood-scoured bedrock testify to the competence of some of these floods in the mobilisation of debris. Large fan/deltas prograding into Crooked Lake must have been formed by truly large floods, far greater than that which occurred during January 1993. The 1993 flood carried some silt, but otherwise bedload and washload was minor. The large size of clasts in the valley-floor debris, and the propensity for stream floors to become armoured with imbricated cobbles, mean that only very large floods are capable of substantial geomorphic work in the proglacial zone. How and when these very large floods formed is uncertain, but they are clearly beyond the realm of the normal summer melt stream: floods released from large, failed ice dams are implied. Proc. LINN. SOc. N.S.W., 119. 1998 D.B. GORE AND J. PICKARD 193 Figure 10. Photo of the flowpath between the upper and middle reaches, at peak flow of 8 m’s'. Photo taken 25 January 1993. Ice dams can fail via a number of mechanisms: seepage, downcutting, undercut- ting, melt, flotation and mass failure (Xia and Woo 1992). At southeastern Vestfold Hills, downcutting during failure of Ice Dams #b and #d, and seepage failure of other small snowbank dams have been observed by Pickard (1986) and Gore (1995:59-61). Melt occurs in all cases, and undercutting may also occur depending on the geometry of the dam and dam outlet. These processes observed in or inferred for Vestfold Hills are simi- lar to those described by Xia and Woo (1992), except that we have no reason to infer dam flotation or mass failure of the dam material. Field observations of seepage failure and downcutting allow us to build a general model of ice dam failure at Vestfold Hills; (1) a seepage zone develops at the firn/ice boundary. (2a) the seepage zone, below the snow surface, becomes a tunnel through which water can flow rapidly, or (2b) the seep- age zone reaches the snow surface and sheet flow develops. Phase 2 may be quite slow, if the snowpack is able to absorb large amounts of water. (3) downcutting follows rapid- ly, with a positive feedback between the discharge volume and rate of downcutting. The outlet through the dam may remain a tunnel (e.g. Fig. 7 at Dam #d), or if the overlying snow lacks the tensile strength to span the gap, an open channel will form (e.g. Dam #b during 1993). This sequence is essentially a simplified version of the model described by Woo and Sauriol (1980), except that at Vestfold Hills, the smaller snowbank dams tend to develop a saturated zone at the upper surface of the dam, while the larger ice dams dis- cussed in this paper were never observed to saturate the upper surface of the dam, but instead developed seepage tunnels. A considerable time lag exists between the first appearance of melt on the ice sheet and the drainage of that melt into the sea at Ellis Rapids (Fig. 1). We have marked on Fig. | our observations of the appearance of first runoff at several locations in the Druzhby System, during the 1980/81 summer. Several points are of interest. First, while Tierney Creek commenced flow during early November 1980, the streams in southeast- Proc. LINN. Soc. N.S.W., 119. 1998 194 PROGLACIAL HYDROLOGY IN EAST ANTARCTICA ern Vestfold Hills (those in Fig. 2) did not start to flow until late December 1980. We believe that the seven week delay in the onset of runoff from southeastern Vestfold Hills was due to retention of runoff behind snowbank and ice dams. Second, despite nearly two months of inflow from Tierney Creek, Crooked Lake had no outflow until early January, 1981. It may be coincidental, but the Crooked Lake outflow followed soon after the southeastern creeks. During the 1980/81 summer, Tierney Creek flowed into Crooked Lake for nine weeks, and the southeastern creeks into Crooked Lake for two weeks, prior to runoff in the Tierney River (Fig. 1). This delay in the onset of outflow of Crooked Lake was because winter and spring ablation had caused the lake to drop below its outlet sill, and the initial inflow refilled the lake basin to the height of the outlet sill. Colbeck (1977) noted a sudden increase in flow discharge at Ellis Rapids during 15-20 January, 1976, which he believed to be due to the start of outflow from Crooked Lake. While our observations support Colbeck’s hypothesis, an alternative interpretation lies with the behaviour of the ice dammed lakes in southeastern Vestfold Hills. Given the modulation of discharge afforded by the ice dams in southeastern Vestfold Hills, it is possible that sudden increases in the discharge hydrograph downstream at Ellis Rapids may simply reflect the release of ice dammed reservoirs. There are no relevant field observations from 1976 to resolve whether or not Colbeck’s hydrograph was affected by ice-dammed reservoir discharge. In any case, the chronology of the onset of drainage in Fig. 1 demonstrates that the individual lake basins do act as retention ponds, that must be filled before overflowing. The streams draining into the eastern end of Crooked Lake are notable because of the changes in flow patterns in different channels. Two changes have been observed and these have important implications for interpreting glaciofluvial deposits should the ice retreat further, and by analogy, understanding the fluvial history of areas that are now distant from the ice sheet. Adamson and Pickard (1986, their Fig. 4.16) showed flow north and east from Lake 5 to Lake 4 and then Lake 2. Here it joined the major drainage route through to Lake 10 and onwards to Crooked Lake. This flow was based on field observations during the 1978/79 and 1980/81 summers. However, when visited during the 1987/88 summer, Gore (1993) found that the ridge of glacier/ice sheet ice between Lakes 5 and 6 had lowered sufficiently to cause Lake 5 to drain to the west. The diver- sion of this drainage meant that the channels draining north through Grimmia Gorge to Crooked Lake effectively captured additional catchment and meltwater. Observations from 1987 to 1993 showed that the small lake between Lakes 5 and 4, which used to form part of the flowpath, is now abandoned. The water level is lowering rapidly, and that lake will soon become a small, dry basin. During the 1992/93 summer, the failure of Ice Dam #b directed water from the ice- dammed lakes, almost directly west to the top of Grimmia Gorge and into Crooked Lake. During that summer, the flowpath from the ice dammed lakes to Crooked Lake via Sickle, Verkhneye and Foot Lakes was deprived of water, and no flow was observed entering Lake Verkhneye. Presumably during the 1993 winter, as the level of Lake 10 was low, snow would accumulate at the site of Ice Dam #b. Once Dam #b reformed, then water would be diverted once again toward the northern flowpath. Conditions favourable for ice dam reformation to occur would be a cold winter with a large snowfall, in con- junction with a following cold summer with minimal meltwater production. Several workers have noted the apparent absence of glaciofluvial features in Vestfold Hills, and inferred the importance of sublimation as a means of deglaciating Vestfold Hills (Hirvas and Nenonen 1991; Hirvas et al. 1993, 1994a, 1994b). The evi- dence presented in this paper demonstrates that fluvial activity occurs in the present day proglacial zone, with stream discharges of at least 8 m’s', and that much larger flows have occurred in the past to form the large fan/deltas shown in Fig. 6. Further from the ice sheet in northwestern Vestfold Hills, the terrain is more subdued and the development of ice dams there during deglaciation would always have been rare. As a consequence, streams Proc. LINN. SOc. N.S.W., 119. 1998 D.B. GORE AND J. PICKARD 195 there would always have been feeble and as a result glaciofluvial features subtle. A further complication is that glaciofluvial features may develop in discontinuous sections. Should the ice sheet retreat sufficiently far that there are no ice or snowbank remnants in south- eastern Vestfold Hills, a bewildering array of discontinuous fluvial features will remain whose history will be impossible to interpret accurately. It would be almost impossible to reconstruct the snow and ice dams, and certainly impossible to detect the changes in direc- tion of flow that we have described here. Thus, the features that we describe are more than of local interest. They provide an explanation of why complete understanding of fluvial features near ice sheet (and even glacier) margins may never be achieved. CONCLUSIONS The drainage system of southeastern Vestfold Hills is dominated by a series of ice-dammed lakes. The lakes are dominantly controlled by three ice dams, two of which (#b and #c) operate in a flip-flop fashion, unpredictably directing floods larger than 1.5 x 10° m* down alternate drainage routes. One of these routes has reaches incised into glacial sediment with some alluvial deposits, while the other has armoured reaches interspersed with lakes and snowfields. While it is not possible to predict which of the drainage routes will be active in any melt season, Ice Dam #b has failed four times in the six seasons from austral summer 1987/88 to 1992/93 (Gore 1995: 62). The channel reaches below Ice Dam #b have changed little through the three floods since the 1989/90 summer, with the exception of minor slumping of the channel walls due to undercutting. Since the release of an ice-dammed impoundment will be observed in the discharge volume downstream, the sudden increase in discharge at Ellis Rapids observed by Colbeck (1977) may simply have been the record of a flood released from a failed ice dam in southeastern Vestfold Hills. The presence of abundant meltwater at the present day, and the erosional and depositional features described in this paper, seems incompatible with the apparent absence of glaciofluvial features in Vestfold Hills described by Hirvas and colleagues (Hirvas and Nenonen 1991; Hirvas et al. 1993, 1994a, 1994b). ACKNOWLEDGEMENTS We thank the Australian Antarctic Division for logistic support, the Antarctic Science Advisory Group and the Joyce W. Vickery Fund for financial support. We thank Andrew Baird and John Webb for assistance with the stream gauging, and Andrew Baird and George Petcopolous for assistance with surveying. REFERENCES Adamson, D.A. and Pickard, J. (1986). Physiography and geomorphology of the Vestfold Hills. In “Antarctic Oasis’ (Ed J. Pickard) pp. 99-139. (Academic Press: Sydney). Bronge, C. (1989). The hydrology of proglacial Chelnok Lake, Vestfold Hills, Antarctica. University of Stockholm, Department of Physical Geography, Research Report 74, 29 pp. Chinn, T.J. and Dickson, R.H.J. (1986). Hydrology and glaciology, Dry Valleys, Antarctica: Annual Report for 1982-83. Report No. WS 1188. Ministry of Works and Development, Christchurch. Colbeck, G. (1977). Hydrographic project, Davis 1976. Antarctic Division Technical Memorandum 66, 44 pp. Gore, D.B. (1991). Examples of ice damming of lakes in the Vestfold Hills, East Antarctica, with implications for landscape development. In ‘Quaternary Research in Australian Antarctica: Future directions’ (Eds D.S. Gillieson and S. Fitzsimons) pp. 37-44. (The Australian Defence Force Academy: Canberra). Gore, D.B. (1992). Ice damming and fluvial erosion in the Vestfold Hills, East Antarctica. Antarctic Science 4(2), 227-234. Gore, D.B. (1993). Changes in the ice boundary of the Vestfold Hills, East Antarctica, 1947 to 1990. Australian Geographical Studies 31(1), 49-61. Proc. LINN. SOc. N.S.W., 119. 1998 196 PROGLACIAL HYDROLOGY IN EAST ANTARCTICA Gore, D.B. (1995). Geomorphological processes and Quaternary sediments of the Vestfold Hills, East Antarctica. PhD thesis, The University of Newcastle (Australia). Gore, D.B., Pickard, J., Baird, A.S. and Webb, J.A. (1996). Glacial Crooked Lake, Vestfold Hills, East Antarctica. Polar Record 32(180), 19-24. Hirvas, H. and Nenonen, K. (1991). Glacial history and Paleoclimates of the Vestfold Hills area, East Antarctica. In “FINNARP-89 Symposium Report No. 1’ (Ed Anon) pp. 31—38. (Ministry of Trade and Industry: Helsinki). Hirvas, H., Nenonen, K. and Quilty, P. (1993). Till stratigraphy and glacial history of the Vestfold Hills area, East Antarctica. Quaternary International 18, 81-95. Hirvas, H., Lintinen, P. and Nenonen, K. (1994a). Properties of till fines in the Vestfold Hills and Vestfjella Areas, Antarctica. In “Antarctic Reports of Finland: FINNARP Symposium Report 4’ (Ed Anon) pp. 20-27. (Ministry of Trade and Industry: Helsinki). Hirvas, H., Lintinen, P. and Nenonen, K. (1994b). Properties of till fines in Finland and Antarctica. Acta Univ. Oul. A 251, 9-23. (In Finnish with English summary) Pickard, J. (1986). Spatial relations of the vegetation of the Vestfold Hills. In “Antarctic Oasis’ (Ed J. Pickard) pp. 275-308. (Academic Press: Sydney). Selby, M.J. (1971). A method of gully erosion, Taylor Dry Valley, Antarctica. New Zealand Journal of Geology and Geophysics 14, 484-485. Tierney, T.J. (1975). An externally draining freshwater system in the Vestfold Hills. Polar Record 17(IIl), 684-685. Woo, M.-K. and Sauriol, J. (1980). Channel development in snow-filled valleys, Resolute, N.W.T., Canada. Geografiska Annaler 62A(1-2), 37-56. Xia, Z. and Woo, M.-K. (1992). Theoretical analysis of snow dam decay. Journal of Glaciology 38(128), 191-199. Proc. LINN. SOc. N.S.W., 119. 1998 A New Genus of Australian Soft Scale Insect (Hemiptera: Coccidae) with Species on Capparis (Capparaceae) and Doryphora (Monimiaceae) from New South Wales PENNY J. GULLAN! AND CHRIS J. HODGSON? 'Division of Botany and Zoology, The Australian National University, Canberra ACT, 0200, Australia, and CSIRO Entomology, GPO Box 1700, Canberra ACT, 2601; and *Environment Department, Wye College, University of London, nr Ashford, Kent, TINZS SAL UK GULLAN, P.J. AND Hopeson, C.J. (1998). A new genus of Australian soft scale insect (Hemiptera: Coccidae) with species on Capparis (Capparaceae) and Doryphora (Monimiaceae) from New South Wales. Proceedings of the Linnean Society of New South Wales 119, 197-217. A new genus, Austrolecanium (Hemiptera: Coccidae: Coccinae), is erected for two species of soft scale from New South Wales. A. sassafras, sp. nov., 1s bright green in life and occurs on the leaves of Doryphora sassafras Endl. (Monimiaceae) in wet gullies in the Blue Mountains and near coastal New South Wales. A. cappari (Froggatt), comb. nov., has been collected from the leaves of Capparis mitchellii Lindley (Capparaceae) in north-western New South Wales. Both species appear to be ovoviviparous and four female instars are known for A. sassafras. The adult female and first-instar nymph of A. cappari and A. sassafras and the second-instar male of A. sassafras are described and illustrated. A key to separate the female instars and the second-instar male of A. sassafras is provided. An apparently predatory cecidomyiid (Diptera) associated with the brood of A. sassafras is reported. Manuscript received 20 August 1997, accepted for publication 19 November 1997. KEYWORDS: Coccidae, Austrolecanium, Platylecanium, Doryphora, Capparis, Cecidomyiidae, taxonomy INTRODUCTION The Australian soft scale insects (family Coccidae), especially endemic species, are poorly known. Other than taxonomic studies of Ceroplastes (Qin and Gullan 1994), Cryptes baccatus (Farrell 1990) and the Pulvinariini (Qin and Gullan 1992), the only well-known species are a few cosmopolitan pests, such as Coccus hesperidum Linnaeus, Parasaissetia nigra (Neitner) and Saissetia oleae (Olivier) (Williams and Watson 1990), and the type species of three Australian genera, Alecanopsis Cockerell, Austrolichtensia Cockerell and Ceronema Maskell (redescribed by Hodgson 1994). Most species cannot be identified, often not even to genus, with the available literature. Froggatt’s (1921) revision of Australian scale insects is the last major taxonomic work to treat the Australian Coccidae as a whole and many of the names and generic concepts used by Froggatt are no longer valid. Recently, some bright green scale insects of an undescribed species of coccid were collected on the leaves of sassafras, Doryphora sassafras Endl. (Monimiaceae). The infested host tree was growing on the edge of a creek in an undisturbed, coastal forest of New South Wales. There are only two species of Doryphora Endl. (Boland et al. 1984) and both are endemic to Australia. D. sassafras occurs in wet forests of the Great Proc. LINN. Soc. N.S.W., 119. 1998 198 A NEW GENUS OF SOFT SCALE INSECT Dividing Range and coastal areas of New South Wales and Queensland (Harden 1990a) and D. aromatica (Bailey) L.S. Smith is confined to rainforests in north Queensland (Lassak and McCarthy 1983). The insect fauna of Doryphora has not been recorded, although levels of insect herbivory of the leaves of D. sassafras have been studied in New South Wales rainforests (Lowman 1985). The features of the adult female of this new species place it in the tribe Paralecaniini of the subfamily Coccinae (as defined by Hodgson 1994), in which it keys out near Platylecanium Cockerell and Robinson, Melanesicoccus Williams and Watson and Neosaissetia Tao and Wong, although it does not closely match any of these genera. Melanesicoccus is known from three species (Ben-Dov 1993): the type species from Papua New Guinea and two species from the Solomon Islands (Williams and Watson 1990; Hodgson 1994). Neosaissetia has four described species (Ben-Dov 1993). The type species, from Taiwan, was redescribed by Hodgson (1994) and the other three species are from Papua New Guinea (Williams and Watson 1990), the Philippines (Morrison 1920) and Malaysia, Singapore and Thailand (Takahashi 1942). Platylecanium has 11 described species (Ben-Dov 1993), also from the Australian and Oriental regions. The only Australian species currently placed in this genus 1s P. cappari (Froggatt), which is known only from the original collection on leaves of wild orange, Capparis mitchellii Lindley (Capparaceae), in north-western New South Wales (Froggatt 1915). Our detailed study of Froggatt’s original material of P. cappari has revealed that it is congeneric with the new species from sassafras. The host plants of these two Australian coccid species are not closely related and occur in quite different vegetation types (Harden 1990a,b), although both have leaves that contain alkaloids (Gharbo et al. 1965; Lassak and McCarthy 1983; Collins et al. 1990). The foliage of C. mitchelli is rel- ished by stock but there are unsubstantiated reports of the plant causing poisoning (Cunningham et al. 1981). The alkaloids of Capparis L. do not appear to have been iden- tified. Both the crushed leaves and the bark of D. sassafras are highly aromatic and have a similar alkaloid composition, with the leaves also containing an essential oil rich in toxic safrole (Gharbo et al. 1965; Chen et al. 1974; Lassak and McCarthy 1983). The use of unrelated host plants by these two coccid species may be associated with their ability to tolerate, or perhaps even respond to, the alkaloids or other compounds. Phloem-feed- ing insects are not protected from the plant secondary compounds of their host plants, as evidenced by the presence of plant-derived compounds, such as alkaloids, in their honey- dew (Dreyer et al. 1985; Molyneux et al. 1990) This paper erects a new genus, Austrolecanium, for the two Australian species dis- cussed above. The coccid from Doryphora is described as a new species and the coccid from Capparis is redescribed and formally transferred to Austrolecanium. The new genus and its two species are described and illustrated based on both the adult females and the first-instar nymphs (crawlers). In addition, the second- and third-instar females are described and the second-instar male is described and illustrated for the species from sas- safras and the pupa of an apparently predatory cecidomyiid also is illustrated. Hodgson’s (1994) key to coccid genera 1s modified to include this new genus and diagnostic fea- tures that readily distinguish the two species of Austrolecanium are listed. MATERIALS AND METHODS The descriptions largely follow the format and terminology of Hodgson (1994). To prepare adult females and nymphs as microscope slide-mounts, body contents were cleared in cold 10% w/v potassium hydroxide (KOH) solution overnight, the cuticle was stained in acid fuchsin in acid alcohol, dehydrated in three changes of absolute ethanol and one of absolute propan-2-o] and then placed in three changes of xylene prior to mounting in Canada balsam. Scanning electron microscopy (SEM) was used to examine Proc. LINN. SOC. N.S.W., 119. 1998 P.J. GULLAN AND C.J. HODGSON 199 the cuticular structures of the first-instar nymphs. Scale insects were prepared for SEM after preservation and storage in lactic-alcohol (Upton 1991) or 80% ethanol. Each speci- men was dehydrated in a graded ethanol series, dewaxed in xylene, rehydrated through a graded ethanol series into distilled water, post-fixed in 1% aqueous osmium tetroxide, washed in distilled water and sonicated briefly to remove any black precipitate, freeze- dried, glued onto a metal stub with nail varnish and coated with gold palladium under vac- uum. Specimens were then examined and photographed using a Cambridge $360 SEM. Each listed scale insect is mounted on a separate microscope slide, unless other- wise specified. All measurements were made on slide-mounted specimens. The following abbreviations are used for depositories: ANIC — Australian National Insect Collection, CSIRO, Canberra ASCT — The Agricultural Scientific Collections Trust, NSW Agriculture, Orange [for- merly the Biological and Chemical Research Institute (BCRI), Rydalmere] BMNH — The Natural History Museum, London. FCNI — State Forests of NSW Insect Collection, Beecroft, Sydney GENUS AUSTROLECANIUM, GEN. NOV. Type species: Austrolecanium sassafras, sp. noy., by present designation. Description of adult female Body oval, 3-10 mm long, 2—7 mm wide, generally slightly asymmetrical with one side of body slightly shorter than other (Figs 1, 4); margins of anal cleft closely adpressed; rather flat. Dorsum membranous or only very slightly sclerotised. Eyespot present, displaced some distance onto dorsum. Dorsal setae present: flagellate or spinose. Three types of dorsal pore: (1) largest, a closed pore with a granulate surface (apparently preopercular pores), most abundant lateral to anal plates, (11) a smaller closed pore with a granular sur- face, frequent elsewhere on dorsum; and (iii) a minute dorsal microductule with a long inner ductule, slightly swollen proximally; also fairly frequent throughout. Dorsal tubular ducts and dorsal tubercles absent. Anal sclerotisation absent; anal plates together approx- imately quadrate, each with 4 small setae near apex; no pores or ridges dorsally on plates; setae on inner margins of ano-genital fold and lateral margins variable. Anal tube usually subequal to anal plates in length; anal ring with 3 or 4 pairs of setae. Margin with spinose marginal setae. Stigmatic clefts distinct, each with a well- defined stigmatic sclerotisation; each cleft with stigmatic spines at inner end. Anal cleft quite deep, sides closely adpressed but not fused. Venter membranous. Ventral setae: with a single pair of long pregenital setae; other pregenital setae short and frequent in bands medially across abdominal segments; other setae most frequent mediolaterally on anal lobe, becoming less so anteriorly; submargin- al setae small or minute; with 1—2 pairs of interantennal setae. Pregenital disc-pores often rather malformed, mostly with 7—9 loculi; most abundant on pregenital segment (VII) but sometimes with a few on abdominal segments III to VI. Spiracular disc-pores mainly with 5 loculi; in fairly narrow bands between spiracles and margin and extending for a short distance medial to spiracle. Preantennal pores absent. Ventral microducts minute, each with outer ductule 2—3 wm long and inner ductule inconspicuous; sparsely scattered throughout venter but concentrated in a U-shaped band surrounding labium. Ventral tubular ducts absent. Spiracles well developed, each with a conspicuous, narrow muscle- plate to peritreme. Legs much reduced, with perhaps 3 segments plus a claw; trochanter Proc. LINN. Soc. N.S.W., 119. 1998 200 A NEW GENUS OF SOFT SCALE INSECT Figures 1—2. Live adult females of Austrolecanium sassafras sp. nov.: 1, two females feeding near the midrib of a leaf of Doryphora sassafras (the paired white streaks on the coccids are due to wax in the stigmatic fur- rows underneath); 2, a female photographed with transmitted light to reveal the internal structures, especially the tree-like tracheal system. Figure 3. Empty test produced by a second-instar male of A. sassafras, showing lines and patches of white wax secreted by the dorsal tubular ducts; the sutures of the test are along the lines. Figure 4. Two dry adult females of Austrolecanium cappari (Froggatt) on a dry leaf of Capparis; four females have fallen off the leaf, leaving behind a few crawlers and piles of cast-off embryonic membranes (arrowed), which appear as white patches. Scale line on each figure = 2 mm. Proc. LINN. SOc. N.S.W., 119. 1998 P.J. GULLAN AND C.J. HODGSON 201 fused with femur, tibia and tarsus also fused but occasionally showing slight signs of a pseudoarticulation; tarsal digitules both finely knobbed and usually slightly longer than claw digitules; claw digitules longer than claw, | broader than other, both finely knobbed; claw small, without a denticle. Antennae reduced, each with about 6 segments, but segmentation often obscure; flagellate setae only present on basal 2 segments. Mouthparts displaced somewhat nearer to shorter side of body, otherwise apparently nor- mal. Labium 2-segmented. Diagnostic features of adult female Adult females of this genus can be identified by the following combination of characters: (1) asymmetry of mouthparts; (11) stigmatic clefts distinct, with a stigmatic sclerotisation and stigmatic spines; (111) pregenital disc-pores generally with 7—9 loculi; (iv) absence of both dorsal and ventral tubular ducts; (v) spinose marginal setae; (v1) reduced, indistinctly-segmented antennae; and (vii) reduced legs. It appears to fall within the Paralecaniini as defined by Hodgson (1994), in which it keys out with Neosaissetia Tao and Wong, but can be easily separated using the following modification to Hodgson’s key to adult female Coccidae: GROUP D: subfamily COCCINAE, tribe PARALECANIINI (Hodgson 1994, pp. 78-79) (NB. Hodgson’s key is based only on the type species of each coccid genus) 8. Claws with a denticle on widest part; with 9-14 stigmatic spines present in a trian- gle situated at base of each stigmatic cleft; stigmatic cleft without stigmatic sclero- tisation; dorsal tubercles absent ............... Melanesicoccus Williams and Watson — Claws each without a denticle on widest part; with 2—3 (occasionally more) stig- matic spines along inner margins of each stigmatic cleft, not arranged in a triangle; stigmatic clefts each with distinct stigmatic sclerotisation; dorsal tubercles present OFA SEIU wie alee cpseeys Risa Meee deter quicken ics asain anol t pais Setrsuien Seaaseu cetera sce 8a 8a Mouthparts placed asymmetrically; dorsal tubercles absent; legs distinctly reduced; claw and tarsal digitules with small apical knobs; pairs of long pregenital setae ony pkeSent oni pregemitalesesmment ee. easy. seer Austrolecanium, gen. nov. — Mouthparts symmetrically placed; dorsal tubercles present in a sparse submarginal ring; legs well developed; claw digitules with broad apical knobs; pairs of long pregenital setae present on posterior three pregenital segments ......................5. Sada Ueno tobe e He Sed See Gane BEE Cem eR Seets OREM CT CE ae nie eae Neosaissetia Tao and Wong Description of first-instar nymph Body of unfed crawler 430-590 um long, 230-330 um wide, rather oval; body becomes wider relative to length after nymph has commenced feeding (see inset AA in Fig. 7); with anal plates only slightly withdrawn into a short, wide anal cleft in youngest specimens but at end of a short anal cleft with adpressed sides in mature specimens (see inset AA in Fig. 7). Dorsum membranous, derm corrugated on young specimens but apparently smooth on mature specimens; with a narrow, distinct, sclerotised border to stigmatic clefts; seg- mentation visible on thorax and abdomen. Eyespot present on margin. Dorsal setae minute (length approximately equal to diameter of basal socket), in 3-4 pairs medially on head and thorax. Dorsal pores of 3 types: (i) a small simple pore in a submarginal line; (ii) a microductule, which is either minute or much enlarged into a figure-of-eight pore, fairly frequent over most of dorsum; and (iii) a pair of minute trilocular pores near anterior margin of head. Dorsal tubular ducts, preopercular pores and dorsal tubercles absent. Anal plates together quadrate, but rather elongate, each with a very long apical Proc. LINN. Soc. N.S.W., 119. 1998 202 A NEW GENUS OF SOFT SCALE INSECT seta, 2 small setae on inner margin and another near apex on posterior margin. Anogenital fold with a single pair of small setae anteriorly and another pair on lateral margins; supporting bars distinct. Anal ring with 3 pairs of setae; anal tube rather short, less than half length of plates. Margin with a row of finely spinose marginal setae, distribution typical of first- instar coccids, with 6 pairs anterior to anterior stigmatic cleft, 2 (rarely 3) setae between anterior and posterior clefts and 8 pairs posteriorly on abdomen; setae on head longest. Stigmatic clefts shallow, each with a distinct stigmatic sclerotisation and 2 stigmatic spines that point posteriorly, 1 spine longer than other; usually with a marginal seta closely associated posteriorly. Venter membranous; segmentation visible on thorax and abdomen. Ventral setae flagellate, distributed as follows: 1 pair of long pregenital setae (occasionally 1-2 more pairs present); | pair of long interantennal setae; all other setae much shorter: | pair per segment in a mediolateral line on abdomen, | pair medially associated with each meso- and metacoxa, and in a sparse submarginal line on each side of body with 7 setae on abdomen, | between stigmatic clefts, 0-1 laterally on prothorax, plus 1 anteriorly on head. Pregenital disc-pores absent. Spiracular disc-pores in a single line of 2—5 pores per stigmatic furrow with none extending medially past peritreme; each pore with 3-7 loculi. With a single pair of ventral microducts, midway between pro- and meso-coxae; very occasionally one present elsewhere. Spiracles small. Legs well developed, each trochanter with a single Jong seta; tibia with a longish seta on ventral margin and 2 other short setae; tarsal digitules dissimilar, | much longer and broader than other, both with knobbed apex except on prothoracic tarsus where | digitule flagellate; claw digitules dis- similar, one broader than other, both shorter than tarsal digitules, each with a small knobbed apex; claw 15-19 um long with a distinct denticle. Antennae well developed, with 6 segments, apical segment rather long; penultimate 2 segments each with a flagel- late seta in addition to a fleshy seta; third segment with 2 long and | short flagellate setae. Mouthparts centrally placed (even on older nymphs) and typical of Coccidae. Diagnostic features of first-instar nymph The most striking features of the crawler are as follows: (i) the stigmatic sclerotisa- tion; (ii) presence of two stigmatic spines, both pointing posteriorly; (iii) presence of dor- sal setae; (iv) dissimilar tarsal digitules; (v) dissimilar claw digitules; (vi) presence of a denticle on claw; (vii) presence of single seta on each trochanter; and (viii) presence of a flagellate seta on penultimate two segments of antennae. Included species The genus Austrolecanium has been erected for two species (A. sassafras, sp. nOV., and A. cappari (Froggatt), comb. nov.), restricted to eastern Australia. For separation of these species, refer to the diagnoses that follow the descriptions of the adult female and first-instar nymph of each species. AUSTROLECANIUM SASSAFRAS, SP. NOV. (FIGS 1-3, 5-7) Material examined Types Holotype, adult female, NEw SOUTH WALES: Yadboro State Forest, off Western Distributor Rd, on Carters Creek, 35°31'00"S, 150°03'20"E, ex leaf of Doryphora sas- safras, 5.vii.1995, P.J. Gullan (ANIC), hereby designated. Proc. LINN. SOc. N.S.W., 119. 1998 P.J. GULLAN AND C.J. HODGSON 203 Paratypes. NEw SOUTH WALES: 13 adult females, same data as holotype (9 ANIC; 1 ASCT, 1 BMNH; 2 FCNI); 2 adult females, 2 third-instar females containing pharate adults, | pharate second-instar female (on slide with first-instar nymphs), 5 second- instar males, 87 first-instar nymphs (10 slides, 5 of which also have cecidomyiid pupae), same data as holotype except 12.1.1996 (ANIC except 1 slide with 8 first-instar nymphs and | cecidomyiid pupa in BMNH); 5 mature first-instar nymphs, same data as holotype except 16.ix.1996 (ANIC); second-instar males (1 slide), same data as holo- type except 28.1x.1996 (ANIC); 2 adult females, same data as holotype except 3.1v.1996, B.A. Melbourne (ANIC); | adult female, Blue Mountains, Nellys Glen, ex sassafras, WWE 699 (ANIC); 2 adult females, ex Doryphora sp., Wollongbar, 7.vili.1989, V.B. Robinson (ASCT). Other material In addition to the slide-mounted type specimens listed above, there are numerous unmounted specimens of adult females and first-instar nymphs both dry and preserved in lactic-alcohol. These are excluded from the type series. Key to instars (excluding prepupal, pupal and adult males) 1. Stigmatic clefts each with 6—12 stigmatic spines of differing lengths ...... adult female — Stigmatic clefts each with 4—5 stigmatic spines, 2 longer and stouter than others ...... ATO EN OIE IE ote NS Onan AERO ROMREH CER rReOTGE ne mEvaly By ntes yd third-instar female — Stigmatic clefts each with 3 stigmatic spines, 2 much longer than third .................. igi fal ith ca ae Se Ree at anye ete MOROR eh 5 e ha ey (second-instar nymph) 2 — Stigmatic clefts each with 2 stigmatic spines, | longer than other ...first-instar nymph 2. Dorsum with tubular ducts arranged in distinct lines.................. second-instar male == DOrsumwithouttubulamductS ss +.08 eee eee eee second-instar female Description of adult female (Figs 1, 2, 5) (measurements based on 10 specimens) Live specimens bright green with shiny dorsum, especially in mature females; young adults partially transparent so that internal organs are visible in transmitted light (Fig. 2). Body oval and usually asymmetrical, 5.4—9.5 mm long, 3.7—7.0 mm wide. Dorsum membranous except for a distinct, clearly marked area of sclerotisation around inner margin of each stigmatic cleft on all specimens, and areas of slight sclero- tisation around body margin and anal plates on old specimens. Eyespot far from body margin, just anterior to level of antennal bases. Dorsal setae flagellate, 15—30 um long, frequent throughout but most abundant and slightly longer near anal plates. Dorsal pores of 2 kinds: (1) minute microductules, | 4m in diameter, with inner ductule tubular and 5—7 ym long at its proximal end, filamentous distally; frequent and fairly evenly distributed throughout; and (i1) simple, closed, pores, 3—5 um in diameter, scattered over dorsum, but largest in a broad area around anal plates; latter, larger pores probably preopercular pores. Anal plates each triangular with anterior and posterior margins subequal, each plate 200-240 um long, 95-120 um wide; with 2 setae apically on each plate plus a small seta and a slightly longer seta posteriorly on posterior margin. Anogenital fold with 2 pairs of setae along anterior margin, approximately of equal length, 35-80 um long; also with a distinct supporting bar on each margin, with 6—10 Proc. LINN. Soc. N.S.W., 119. 1998 204 A NEW GENUS OF SOFT SCALE INSECT Figure 5. Adult female of Austrolecanium sassafras sp. noy. Unless otherwise stated, the letters on this and the following figures refer to the following structures: A, dorsal trilocular pore; B, dorsal seta; C, dorsal microduc- tule; D, dorsal simple pore; E, dorsal tubular duct; F, anal plates and anogenital fold; G, stigmatic area; H, mar- ginal spinose seta; J, spiracular disc-pore; K, pregenital disc-pore; L, ventral microduct; M, complete metatho- racic leg or part thereof; N, tarsus and claw of prothoracic leg; Q, complete antenna. In addition, unless other- wise stated, the scale lines against each vignette are as follows: anal plates (F), stigmatic cleft (G), leg (M) and antenna (Q) = 50 um; all other structures = 5 ym. Proc. LINN. SOC. N.S.W., 119. 1998 P.J. GULLAN AND C.J. HODGSON 205 setae (40-75 um long) in a line on either side, plus 3 pairs of setae more posteriorly. Anal tube slightly shorter than length of anal plates; anal ring 70-83 um in diameter, with 4 pairs of setae, each 225—275 um long. Margin with setae finely spinose, + straight, shorter than dorsal setae, 12—20 um long, in a single marginal row but absent from stigmatic and anal clefts; with 4-11 setae on each side between lateral stigmatic clefts; anal lobe setae not differentiated. Each stig- matic cleft rather “mushroom-shaped’, i.e. with narrow cleft broadening into a wide space; each with a well-defined area of sclerotisation along inner margins of cleft; each cleft with 6—12 stigmatic setae; each seta 13-50 um long, parallel-sided, with a rounded, often slightly clavate apex. Venter with segmentation only visible medially on thorax and abdomen. Ventral setae flagellate, sparsely scattered but longest and most abundant mediolaterally on anal lobes; pregenital segment (VII) with a single pair of long pregenital setae, 115-140 um long, and a group of 8-12 shortish flagellate setae just anterior to each group of pregenital disc-pores; other setae medially on abdomen, with pairs per segment: VI 7-19; V 7-10; IV 7-10; II 9-11; Hf 10-11; also with 1-2 pairs of interantennal setae; submar- ginal setae sparse. Pregenital disc-pores 4-6 wm in diameter, with mostly 7—9 loculi, sometimes slightly misshapen, in two groups of 25-40 pores on either side of ano-genital fold and occasionally with 0-2 on either side mediolaterally on previous segment. Each stigmatic furrow with a narrow band of spiracular disc-pores, each with 3—5 (mostly 5) loculi and 3—6 wm in diameter, with 45—160 pores between spiracle and stigmatic cleft; each band extending a short distance (up to 8 pores) medially past peritreme. Ventral microducts as in generic description, but also frequent in a broad submarginal band. Spiracles: anterior spiracle plus peritreme 115-150 um long, 80-110 um wide; posterior spiracle plus peritreme 110-160 um long, 80—120 wm wide. Legs each 70-140 um long with forelegs shortest; tarsal digitules 15—28 um long, narrower than claw digitules, | broader than other, both with minute apical knobs; claw digitules 10-20 um long, rather parallel-sided, each with a minute apical knob. Antennae each with about 6 indistinct seg- ments; total length 140-170 um; all setae on terminal 3 segments more-or-less fleshy, 8-28 um long. Mouthparts positioned distinctly nearer shorter margin of body. Clypeolabral shield 170-220 um long, 160-190 wm wide. Labium 70-80 wm long, 100-120 um wide. Diagnosis of adult female The adult female of A. sassafras differs from that of A. cappari (Froggatt) comb. nov. (redescribed below) in (i) having many more stigmatic setae per stigmatic cleft (6-12 cf. 1-4 in A. cappari); (11) marginal setae shorter than dorsal setae; (111) dorsal setae flagellate and rather concentrated around anal plates; (iv) ano-genital fold with a line of 6-10 setae along the lateral margins of each supporting bar, and (v) 4 pairs of setae in anal ring. Third-instar female (measurements based on 2 specimens each containing a pharate adult) Body oval, 3.0—3.2 mm long, 2.2—2.3 mm wide. Dorsum membranous except for sclerotised stigmatic clefts. Anal plates each elon- gate triangular, rounded laterally, 150 um long, 60-72 um wide. Anal ring c. 50 um in diameter, with 3 pairs of setae, each 140-175 um long. Margin with spinose setae, 10-14 um long, in a single marginal row. Each stig- matic cleft distinct with a well-defined area of sclerotisation along inner margins of cleft and with 4—5 stigmatic spines (2 longer and stouter than others), each parallel- sided with a blunt apex and a broad basal socket; 2 stouter stigmatic spines 28—40 wm long, 2—3 thinner spines 15—25 um long. Proc. LINN. Soc. N.s.W., 119. 1998 206 A NEW GENUS OF SOFT SCALE INSECT Venter membranous. Stigmatic furrows each with band of spiracular disc-pores (bands difficult to discern because of pharate adult cuticle beneath), each pore mainly with 3-5 loculi and perhaps 20-25 disc-pores in each furrow. Spiracles well developed, with narrow muscle plate associated with each peritreme; length of each spiracle plus peritreme 70-85 wm long, width of peritreme 40-45 um. Legs reduced, 90-100 um long, indistinctly 3-4 segmented due to partial fusion of tibia and tarsus. Antennae reduced, with 6—7 segments but segmentation indistinct; total length 120-130 um. Second-instar female nymph (measurements based on one partially pharate specimen only) Body oval, 1.3 mm long, 0.9 mm wide. Dorsum membranous except for sclerotised stigmatic clefts. Anal plates each elon- gate triangular and rounded laterally, each plate 90 wm long, 32 wm wide. Anal ring 35 yum in diameter, with 3 pairs of setae, each 105—140 ym long. Margin with spinose setae, c. 10-12 ym long, in a single marginal row. Each stig- matic cleft distinct, with a well-defined area of sclerotisation along inner margins of cleft and with 3 stigmatic spines (2 long, | short), each parallel-sided with a blunt apex and a broad basal socket; spines 27-31 um, 36-40 wm and 5—11 ym long, respectively. Venter membranous. Stigmatic furrows each with band of spiracular disc-pores + 1| pore wide, each mainly with 5 loculi, 3-4 wm diameter, with 13-15 disc-pores in each furrow plus 1—2 disc-pores extending a short distance medially past peritreme. Spiracles well developed, with narrow muscle plate associated with each peritreme; length of each spiracle plus peritreme 50-60 wm long, width of peritreme c. 25 um. Legs and antennae not preserved well enough to describe. Second-instar male nymph (Figs 3, 6) (measurements based on 7 specimens) Body oval, 1.9-2.6 mm long, 1.2-1.7 mm wide, showing only slight signs of asym- metry; stigmatic clefts distinct, inner margins sclerotised; anal cleft closely adpressed. Dorsum membranous except for a distinct, clearly marked area of sclerotisation around inner margin of each stigmatic cleft. Eyespot not discerned. Dorsal setae apparent- ly absent. Dorsal pores of 2 kinds sparsely distributed throughout: (1) a minute microduc- tule possessing an inner ductule with balloon-like proximal end (4-5 mm long) and long filamentous distal end (10-13 um long); and (ii) a simple pore about 2—3 wm in diameter. Preopercular pores and dorsal tubercles absent. Dorsal tubular ducts present in a distinct reticulate pattern, with 2 medial lines of ducts extending anteriorly from anal plates to about dorsad to mouthparts and with 4 pairs of lines radiating from medial lines laterally to margin: | pair on abdomen, | pair to each stigmatic cleft and | pair on head; in addition there are submarginal groups of tubular ducts associated with each anterior 3 lateral lines plus a further 3 groups each side anteriorly on abdomen; each tubular duct with a stout outer ductule (15—22 um long), a shorter (7-10 ym), stout, inner ductule and well-devel- oped terminal gland. Anal plates triangular, with anterior margin slightly shorter than pos- terior margins, each plate 88-100 um long, 38-41 wm wide; with 2 apical setae 20-35 um long, plus a short seta posteriorly on posterior margin and another long seta posteriorly on inner margin. Anogenital fold with 2 pairs of setae along anterior margin, length 20-30 um, plus | short seta on each lateral margin of anal cleft. Anal tube subequal to length of anal plates; anal ring 35-39 um in diameter, with 3 pairs of setae, each 110-140 ym long. Margin with straight and rather sharply spinose setae, 9-13 um long, in a single marginal row but absent from stigmatic and anal clefts; with 3—6 setae laterally between stigmatic clefts; anal lobe setae not differentiated. Each stigmatic cleft quite deep, with 3 stigmatic spines; each spine parallel-sided with a blunt apex: 2 longer spines (21-40 um Proc. LINN. SOC. N.S.W., 119. 1998 P.J. GULLAN AND C.J. HODGSON 207 Figure 6. Second-instar male of Austrolecanium sassafras sp. nov. Lettering and scale lines as in Fig. 1, except that the scale line for the stigmatic cleft (G) = 25 um. Proc. LINN. Soc. N.S.W., 119. 1998 208 A NEW GENUS OF SOFT SCALE INSECT long) anteriorly and | much smaller spine (5—15 um long) posteriorly, set in a well- defined area of sclerotisation along inner margins of cleft. Venter with derm membranous; segmentation only visible on abdomen. Ventral setae flagellate, distributed as follows: with a single pair of pregenital setae, 55-60 um long, and 1-2 pairs of moderately long (20-28 um) interantennal setae; all other setae much shorter (3—5 wm long): | pair medially and | pair mediolateral on each abdominal segment; 1—2 pairs associated with each coxa; also in a sparse submarginal band, with probably 1-2 laterally between stigmatic clefts (although none visible); plus a few scat- tered on head and thorax. Pregenital disc-pores absent. Stigmatic furrows with band of small spiracular disc-pores + one pore wide, each with 3—7 (mostly 5) loculi and 4-5 um in diameter, with 9-21 disc-pores in each furrow; also with 0-2 disc-pores extending a short distance medially past each peritreme. No preantennal pores detected. Ventral microducts 5—6 ym long, present in a sparse, broad submarginal band and also in a U- shaped band posterior to labium. Ventral tubular ducts absent. Spiracles well developed, with narrow muscle-plate associated with each peritreme; anterior spiracle plus peritreme 52-62 um long, 25—28 zm wide; posterior spiracle plus peritreme 50-60 um long, 25-30 uum wide. Legs much reduced, with trochanter + fused to femur and tibia fused to tarsus but with a pseudoarticulation, rather stout; total length 85—110 um; tarsal digitules much reduced, of variable length (5-16 wm) and only longer of 2 digitules with slight apical swelling; claw digitules 5-11 um long, usually shorter than tarsal digitules, barely longer than claw and sometimes | digitule with slight apical swelling; each claw small, without denticle. Antennae reduced, probably 7-segmented; total length 115-135 um; all setae on terminal 3 segments fleshy, flagellate setae only on basal 2 segments. Mouthparts posi- tioned distinctly nearer one foreleg. Clypeolabral shield 90-120 um long, 100-110 um wide. Labium 2-segmented, 50-60 um long, 60-70 um wide. Diagnosis of second-instar male nymph The second-instar male is easily distinguished from immature females by its reticu- late pattern of tubular ducts on the dorsum. These ducts apparently produce a whitish wax which is visible as lines on the empty test (Fig. 3). First-instar nymph (Fig. 7) (measurements based on 10 specimens) Live specimens with bright yellow body. Body oval, 430-570 um long, 230-310 um wide. Dorsum with dorsal setae 2-4 um long. Dorsal pores consisting of: (i) simple pores, 2 wm in diameter, in a submarginal line extending from head to posterior abdomen; (11) minute microductules each possessing an inner ductule with a fairly broad proximal end (4—5 ym long) and a filamentous distal end, present in a submarginal line and also medially on thorax and abdomen; and (i11) a pair of small trilocular pores near anterior margin on head. Anal plates each 60-65 um long, 25—28 um wide; each with apical seta 260-300 um long. Anal ring 22—25 um in diameter, with setae 45-80 um long, most ventral pair shortest and thinnest. Margin with setae, 7-20 um long, each slightly bent posteriorly. Stigmatic clefts pre- sent but shallow, each with a distinct stigmatic sclerotisation and 2 stigmatic spines on anterior margin of cleft; 1 spine about 4rd—/rds as long as other (lengths 8—22 um and 29-40 jum, respectively); usually with a marginal seta within sclerotised margin posteriorly. Venter with pregenital setae 38-55 um long, interantennal setae 30-50 um, all other setae 2-4 ym. Spiracular disc-pores each 2—3 ym in diameter and with 3—5 (usually 3) loculi; 3—5 (usually 4) pores in anterior furrows and 3-4 (almost always 4) pores in posterior furrows. Occasionally a ventral microduct just posterior to each scape in addi- tion to one between fore- and midlegs. Spiracles: length of each muscle-plate plus per- Proc. LINN. SOC. N.S.W., 119. 1998 P.J. GULLAN AND C.J. HODGSON 209 Figure 7. First-instar nymph (crawler) of Austrolecanium sassafras sp. nov.; inset (AA) shows the shape of the expanded body of the nymph after it has started feeding. Lettering and scale lines as in Fig. 1, except that the scale lines for the anal plates (F) and the stigmatic cleft (G) = 25 um. _ Proc. LINN. Soc. N.S.W., 119. 1998 210 A NEW GENUS OF SOFT SCALE INSECT itreme 15—22 ym; width of peritreme 8—11 wm. Legs as in generic description. Antennae 160-190 um long; on terminal segment: apical seta 33-44 um long, longest flagellate seta 90-110 um long. Clypeolabral shield 80—90 wm long, 67-80 wm wide. Labium 34-38 um long, 45—60 um wide. Diagnosis of first-instar nymph The first-instar nymph of A. sassafras differs from that of A. cappari primarily in possessing minute and scarce dorsal microductules, whereas in the latter species they are much more frequent and enlarged into “figure-of-eight’ pores. Comments and biological notes Among Froggatt’s dry collection in ANIC are three drymounts of A. sassafras with the manuscript name of ‘“Lecanium johnstoni’. Apparently, this proposed name was to honour the collector. Froggatt’s accession notebook records specimens of the species under that name to be from Doryphora sassafras at Katoomba, collected on 20.vi.1916 by Dr S. Johnston. Adult females of A. sassafras from the single infested tree at the site in Yadboro State Forest occurred mostly near veins on the underside of leaves (Fig. 3). Their honey- dew, which had fallen on the foliage below, had led to the growth of black sooty mould. Females turned brown to black-brown after death. In early January 1996, there were first- instar nymphs under the bodies of adult females and newly-settled nymphs on the leaves, mostly on the upper surfaces, of the same tree. However, mature first-instar nymphs also were collected from both abaxial and adaxial leaf surfaces of the same tree in September 1996, suggesting that emergence of crawlers may occur any time from spring to summer or that two cohorts are present at the site. This species is apparently ovoviviparous. A pharate second-instar female nymph was collected in January 1996. Two dead adult females pharate in their third-instar cuticle were collected in January 1996, confirming that this species has four female instars. It was not possible to illustrate or fully describe the second- and third-instar females due to the difficulty of seeing cuticular features on the few specimens available. Several dead second-instar males were obtained from under tests (Fig. 3) on the upper surface of leaves; adult males probably are present in early summer. Amongst the coccid progeny collected from under the female coccids were some tiny pupae of an unidentified species of Cecidomyiidae (Diptera) (Fig. 8). Pupae of only a very few Australian species of cecidomyiid have been described and none of them have been associated with coccids or Doryphora (P. Kolesik, pers. comm.). The cecidomyiid is probably an undescribed species (P. Kolesik, pers. comm.) and the larvae may be predatory or endoparasitic on the coccid progeny (Harris 1968). AUSTROLECANIUM CAPPARI (FROGGATT), COMB. NOV. (FIGS 4, 9, 10) Lecanium cappari Froggatt 1915: 604. — Froggatt 1921: 29. Platylecanium cappari. — Ben-Doy 1993: 237. Material examined Types Lectotype, adult female, NEW SOUTH WALES: Gunnedah, ex Capparis [as ‘Cappras’ on label], WWF481 (ASCT), hereby designated. Paralectoypes. NEW SOUTH WALES: 3 adult females, | drymount with additional 2 adult females, same as data as lectotype (ASCT). [The dry females from Froggatt’s Proc. LINN. Soc. N.S.W., 119. 1998 P.J. GULLAN AND C.J. HODGSON 211 Figure 8. Pupa of an unidentified species of Cecidomyiidae (Diptera) found amongst the coccid crawlers under the abdomen of an adult female of A. sassafras. Letters refer to the following structures: S, abdominal dorsal spine (large medial projection) or spicule (small ventral or dorsolateral projection); X, lower part of face; Y, prothoracic spiracle with trachea ending at apex; Z, abdominal spiracle. Proc. LINN. SOC. N.S.W., 119. 1998 PN A NEW GENUS OF SOFT SCALE INSECT collection are included as paralectotypes because Froggatt based his descriptions on unmounted material and thus all of his specimens must be considered as syntypic.] Other material examined NEW SOUTH WALES: 7 adult females, 77 first-instar nymphs (8 slides), 3 dry- mounts with additional 3 adult females and first-instar nymphs, Nyngan, ex Capparis, 11.x1.1921, WWE 1061 (ANIC); 2 adult females, 56 first-instar nymphs (4 slides), 3 dry- mounts with additional 19 adult females and first-instar nymphs, same data as ANIC specimens (ASCT); 4 adult females (1 slide), Moonie River via Collarenebri, ex Capparis mitchelli, Oct 1911, S.W. Jackson (ASCT). All specimens from Nyngan were collected in 1921 and, because the species was described in 1915, cannot be part of Froggatt’s syntypic material. The label for the type specimens from Gunnedah does not have a date (see comment under Biological notes). Description of adult female (Fig. 4, 9) (measurements based on 10 specimens) Live specimens (according to Froggatt 1915) dark chocolate brown with lighter- coloured edges to body when mature; of lighter coloration with yellow body margin when immature. Body oval and usually asymmetrical, 2.9-5.8 mm long, 2.1-4.5 mm wide. Dorsum with membranous derm except for narrow, heavily sclerotised crescents around margin of each stigmatic cleft and slight sclerotisation posterior to anal plates on older specimens. Eyespots displaced from margin, just anterior to level of antennal bases. Dorsal setae bluntly spinose, tending to clavate especially on posterior abdomen, 7—13 um long, sparse throughout but most frequent in a broad submarginal band; scarce medially. Dorsal pores or pore-like structures of 3 kinds: (1) minute microductules, | um in diameter, with inner ductule 10-13 um long, broadly tubular proximally, filamentous distally, fre- quent and fairly evenly distributed throughout; (ii) simple, closed ‘pores’ (probably areas of thin cuticle), 2.5—5 um in diameter, in a broad submarginal band and apparently absent medially; and (111) preopercular pores, flat with a slightly granulate surface, irregularly oval to circular, 4~7 um diameter, in a group on either side of anal plates and extending anterior- ly in 2 sparse, broad, diverging lines to about prothorax. Anal plates each triangular, with anterior margin slightly shorter than posterior margins, each plate 153-172 um long, 70-78 jum wide; with 2 setae apically on each plate, a slightly longer seta posteriorly on inner margin and a shorter seta posteriorly on posterior margin. Anogenital fold with 0-1 pair of setae along anterior margin, 15—25 wm long; with | pair setae at posterior end of lateral margins of anal cleft, 12-15 um long. Anal tube subequal or a little longer than length of anal plates; anal ring 60-73 um in diameter, with 3 pairs of setae, each 175-250 um long. Margin with bluntly spinose setae, each generally bent, with a slightly clavate apex; distinctly longer than dorsal setae, 17—33 um long, in a single marginal row but absent from stigmatic clefts and anal cleft; with 11—24 setae laterally between stigmatic clefts; anal lobe setae not differentiated. Each stigmatic cleft with a well defined area of sclerotisation around inner margins; each cleft usually with 2 (rarely 1, 3 or 4) stigmatic spines; anterior stigmatic spine 28-44 um long, posterior spine 38-50 um long, both spines parallel-sided with a rounded apex. Venter with segmentation only visible on abdomen. Ventral setae flagellate, 7-38 uum long, very sparse on head and thorax, most abundant medially on abdomen and espe- cially mediolaterally on anal lobes where longest; pregenital segment (VII) with a single pair of long (110-140 um) pregenital setae and paired lateral groups of 6—12 short flagel- late setae plus 1—3 setae of intermediate length; other setae medially on abdomen, with pairs per segment: VI 6-12; V 6-10; IV 2-4; II 4~7; II 3; also with | pair of interanten- nal setae; submarginal setae very sparse. Pregenital disc-pores rather misshapen, usually Proc. LINN. SOC. N.S.W., 119. 1998 P.J. GULLAN AND C.J. HODGSON 21 Ww SOOT \ NNere \ Ni ae Figure 9. Adult female of Austrolecanium cappari (Froggatt). Lettering and scale lines as in Fig. 1, except that the scale line for the stigmatic cleft (G) = 25 um. Proc. LINN. SOC. N.S.W., 119. 1998 214 A NEW GENUS OF SOFT SCALE INSECT with 7-9 loculi, 3—6 zm diameter, present as follows: pairs per segment: VII 4-9; VI 4-11; V 0-4; IV 0-4; III 0-1; II 0. Each stigmatic furrow with a band of 30-62 spiracu- lar disc-pores extending from spiracle to stigmatic cleft, each band about 1—3 pores wide, each disc-pore 3—5 zm diameter and usually with 5 loculi; each band with 1—7 disc-pores extending a short distance medially past peritreme. Ventral microducts as in generic description. Spiracles: anterior spiracle plus peritreme 45—60 um long, 42-50 um wide; posterior spiracle plus peritreme 50-70 4m long, 42-55 um wide. Legs each 50-85 um long; tarsal digitules 10-22 ym long, | digitule longer and broader than other, both nar- row with minute apical knobs; claw digitules 7-16 um long, parallel-sided, with only slight apical knobs. Antennae each with about 6 indistinct segments; total length 110-145 sm; with all setae on terminal 3 segments fleshy, 7-19 um long. Mouthparts generally positioned slightly nearer shorter margin of body. Clypeolabral shield 140-160 um long, 145-160 um wide. Labium 55—70 xm long, 90-110 um wide, sometimes twist- ed through 90. Diagnosis of adult female The adult female of A. cappari can be separated from that of A. sassafras in pos- sessing the following features: (i) usually only 2 stigmatic spines per stigmatic cleft; (11) marginal setae longer than dorsal setae; (iii) dorsal setae spinose and rather concentrated in a broad submarginal band; (iv) ano-genital fold with only | pair of small setae on lat- eral margins of each supporting bar, and (v) 3 pairs of setae in anal ring. First-instar nymph (Fig. 10) (measurements based on 10 specimens) Body oval, 450-590 um long, 290-330 ym wide. Dorsum with dorsal setae about 2 um long. Dorsal pores consisting of: (i) simple pores, 2~3 ym in diameter, present in a submarginal line extending from head to posteri- or abdomen and occasionally elsewhere; (ii) microductules enlarged into ‘figure-of- eight’ pores (greatest width of pore 5-6 ym), each possessing an inner ductule (15-22 um long) with a broad proximal end (6-8 zm long) and a longer filamentous distal end, present in segmental bands of about 12 pores across each abdominal and thoracic seg- ment and also + randomly on head; and (iii) a pair of small trilocular pores near anterior margin on head. Anal plates each 42-50 wm long, 20-25 zm wide; each with apical seta 160-210 um long. Anal ring 20-25 ym in diameter, with setae 45-50 um long. Margin with setae, 6-18 um long, each distinctly bent posteriorly. Stigmatic clefts clearly present in older specimens, less obvious in young specimens; each stigmatic area with a distinct stigmatic sclerotisation and 2 stigmatic spines; more anterior spine slight- ly more than half length of posterior spine (lengths 5-10 wm and 8—16 um, respectively); usually with a marginal seta very close to but not within posterior margin of stigmatic sclerotisation. Venter with pregenital setae 35-45 um long (a few individuals also with single long setae on segments VI and V), interantennal setae 20-30 ym long, all other setae 3-8 um long. Spiracular disc-pores each 3-5 ym in diameter and with 3, 5 or 7 loculi; 3-6 pores in anterior furrows and 4 pores in posterior furrows. Spiracles: length of each mus- cle plate plus peritreme 16-20 um; width of peritreme 7-10 ym. Legs as in generic description. Antennae 180-190 wm long; on terminal segment: apical seta 30-38 um long, longest flagellate seta 40-53 um long. Clypeolabral shield 90-95 um long, 80-95 um wide. Labium 30-38 um long, 50-65 zm wide. Diagnosis of first-instar nymph The first-instar nymph of A. cappari differs from that of A. sassafras mainly in having much enlarged dorsal microductules that resemble “figure-of-eight’ pores. Proc. LINN. SOC. N.S.W., 119. 1998 P.J. GULLAN AND C.J. HODGSON i) n Figure 10. First-instar nymph (crawler) of Austrolecanium cappari (Froggatt). Lettering and scale lines as in Fig. 1, except that the scale line for the prothoracic claw (N) = 25 um and the stigmatic cleft (G) = 5 um. Proc. LINN. Soc. N.S.W., 119. 1998 216 A NEW GENUS OF SOFT SCALE INSECT Biological notes The first-instar nymphs of A. cappari were obtained from small aggregations (Fig. 4) under the abdomen of several dry adult females that were attached to leaves and from inside the body cavity of the mothers. A. cappari is clearly ovoviviparous. Crawlers were found under most of the adult females collected in November 1921 at Nyngan. There were no crawlers under the adult females from Gunnedah; the date of collection of these latter specimens is not given on the labels, but from the sequence of entries in Froggatt’s accession notebook (housed in ASCT), they appear to have been obtained in mid-1910. No other immature stages are present in Froggatt’s collection, although he referred to the male tests as being *... white, semi-transparent, elongate oval, flattened, with a white line on either side, converging to a point at the posterior angle. Lateral plates finely crenulate on the margins. Length, “3 of an inch [=c. 2 mm].’ (Froggatt 1915, p. 605). Only the host-plant genus, Capparis, is recorded on Froggatt’s original material. However, both Froggatt’s original description and his accession notebook record ‘Capparis mitchelli’ [sic] as the host plant for both the Gunnedah and Nyngan collec- tions. Furthermore, Froggatt’s dry material consists of whole leaves bearing coccids and these leaves match those of C. mitchellii, as illustrated by Harden (1990b). C. mitchellii, which is known commonly as wild orange or native orange, is found in all mainland states but especially in inland northern New South Wales (Cunningham et al. 1981; Harden 1990b). ACKNOWLEDGEMENTS We thank Peter Gillespie (NSW Agriculture) for access to Froggatt’s specimens held at the Agricultural Scientific Collections Trust and Peter Kolesik (Waite Institute, University of Adelaide) for confirming the iden- tity of the cecidomyiid pupa and providing some information on cecidomyiids. Aimorn Stewart kindly prepared specimens for SEM and also printed the photographs used in Figures 14. We acknowledge the facilities and technical support provided by the Electron Microscopy Unit of the Australian National University. 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Vaan te cash ssi iati BAO sellin y | Weide oa VOR Reaaicny Sey Hy Pua pm iihie fe We Ernst Wi orm PS mE ee Ws PA ES ating | tage’ Pi | is mi y PTR TK Ppa diye Peed ctor Vaile aie, oy) aot wae Peta Si bas sleet heat mp hea ares ¢ raentst i wala Oe een iy ne co Oy PP peti Cosh ba wh} yi mer) Kanifegemere # eNitocee, Val is : = a ray P re 4 ' WU Vi i i Tey 5 a eee. 2 OSE, rv whieh ipinte tani iy Gr wy ; nes: Ni co lie ie 24% Ary TR OTe TE oe ays Or AMSSrTFeY ait Wry dota Ties ry oe . o ee| Mii deal Wabi auiired Susi st f vertices fez wide DE NAS. 04s: = Viele ae a) aS ee sc i Cs pee, dyes fiMasgiall) cmon ly ky sce ih biker: — co. oe ras : qe pie? eo eta Bi aes. ee) as eto we a eee CSO Goeth &, ayaa oe CT ee gah fener hn ia ) “l Aneiatie, wee ef le Sey —= ) as .ak : me. a tae Sere) Wwe Hef) Ane ate yp eure OF ites Ser ee ef tay! salyh oar ism Pri Meyees. Pemhirgeil it Ape ibn, Mee a tek S52 FANG Lae ae D hes WAKE Pe eal ih, fom, O08: cab sdlvadiastta ht LUGO A LED eee Dompboed Td i Mlaiek he ate) Ay a Wight, des Meares Lani a, ob hg Whaley ROW get, Mya ill ay, A Yeap nt Flee’ a, a —— fat iy. Sens) ha Fog) Maman Yipee $ hie anlity Fen Kinad sje! ae rayne Fe A Possible Bioherbicide for Avena fatua L. (Wild Oats): Isolate Collection and Host Range Testing SHANE D. HETHERINGTON, BRUCE A. AULD AND HEATHER E. SMITH Orange Agricultural Institute, NSW Agriculture, Forest Road, Orange, Australia HETHERINGTON, S.D., AULD, B.A. AND SMITH, H.E. (1998). A possible bioherbicide for Avena fatua L. (Wild Oats): Isolate collection and host range testing. Proceedings of the Linnean Society of New South Wales 119, 219-225. A survey of the diseases of wild oats was carried out in NSW during 1995 and 1996. This survey identified a variety of fungal pathogens. Because of its virulence in laboratory tests and its impact in the field, Drechslera avenacea was chosen as the basis of a potential bioherbicide against Avena fatua. A variety of cultivars of wheat, barley and oats were inocu- lated with 12 diverse isolates of this fungus and the disease reaction assessed in each case. In general, Avena fatua was most susceptible to the pathogen, followed in order of susceptibility by oats, barley and wheat. An isolate’s suitability for incorporation into a bioherbicide was based upon the severity of the disease interaction (measured as percentage leaf necrosis ten days after inoculation). Isolates with good potential caused severe symptoms on A. fatua and were less virulent on cereal cultivars. Of the tested isolates, two provided good selectivity and were chosen for further study. As a result of the large number of stored isolates it is possible that other selective isolates may be found. Manuscript received 14 October 1997, accepted for publication 19 November 1997. KEYWORDS: Avena fatua, bioherbicide, Drechslera avenacea, host range, survey. INTRODUCTION Wild oat, Avena fatua L. is a serious weed of a wide range of crops. These include both dicotyledons (e.g. peas, beans, potatoes, sunflower) and cereal crops (Holm et al. 1977). In Australia, wild oats are principal weeds of cereals including wheat, barley and oats. Its effect on wheat is through competition for light (Rooney 1991), nutrients (Kirkland 1993) and also through allelopathy (Perez and Ormeno-Nunez 1991). Herbicide resistant populations of Avena spp (Powles and Holtum 1992) have been recorded in Australia. Hence, calls for augmentation of chemical controls and the diver- sification of control strategies have been made (Howat 1987). The advantages of biological control in reducing use of chemicals, non-target dam- age and environmental contamination are well known. Application of pathogenic organ- isms to grass weeds remains a scarcely investigated option for weed control (Evans 1991). Several attempts have been made to develop bioherbicides effective against wild oats. In Canada, a bioherbicide was developed based on the fungal pathogens Phytophthora palmivora and Colletotrichum gloeosporioides f.sp. aeschymene (sic.) (Mortensen 1983). Pyrenophora semeniperda, a fungal pathogen capable of killing seed, can reduce germination, emergence and vigour of wild oats and is also the subject of bio- logical control research (Medd and Campbell 1996). Recently attention has been given to Drechslera avenacea (Curtis and Cooke) Shoem (teleomorph: Pyrenophora chaetomioides). Wilson (1987) found that D. avenae (Eidam) Scharif [a synonym of D. avenacea; (P.M. Kirk, in litt.)] reduced seedling root and shoot growth by 15% while not affecting winter wheat. However, in this case, the Proc. LINN. SOc. N.S.W., 119. 1998 220 POSSIBLE BIOHERBICIDE FOR AVENA FATUA competitive ability of the weed was unaffected. In China, isolates of D. avenacea have been collected from naturally infected wild oats in wheat fields, cultured and pathogenic- ity tested. The fungus infected wild oats but not wheat (Zonjian and Yanghan 1996). A. fatua is widely distributed across a range of environments and climates; addi- tionally variable phenotypes have been recorded (Whalley and Burfitt 1972). Survey and collection of pathogenic fungi across the geographic and physical ranges of the weed will potentially yield fungal isolates which will be useful as the principal component of a bio- herbicide. A. fatua is also closely related to the cereal crops, notably cultivated oats (A. sativa L). The susceptibility of these crops to the biological control agent must be assessed in the context of wheat/pasture rotations. Our objective in this study was to find suitable endemic fungi as the basis of a bioherbicide. We made no assumptions as to the suitability of candidates before the pro- ject, preferring to base our judgement on preliminary pathogenicity tests of all fungi collected. Based on the severity of symptoms observed in the field and following artifi- cial inoculation, D. avenacea was chosen as the most likely candidate for development as a bioherbicide. MATERIALS AND METHODS Survey of the diseases of wild oats in New South Wales Diseased wild oats (Avena spp) were collected from the field during 1995 and 1996. Plants were collected from roadsides, cultivation (principally wheat) and fallow paddocks. Survey dates were chosen so that plants of various ages could be collected. The area chosen for surveys was based largely on the NSW wheat belt bounded in the north-west by Bourke, the north-east by Glen Innes, the south-west by Hay and Deniliquin and the south-east by Cootamundra and Wagga Wagga. The survey also included several peripheral areas including the Grafton area and semi-rural areas west of Sydney. At each site whole diseased plants were collected and stored at 4°C until they were returned to the laboratory. Where possible, the collection included an inflorescence to confirm host identification; where seedlings were collected this was not possible. Lesions from diseased plants were photographed, assigned a code number, excised and cut into three portions before isolations took place. The portions were soaked in 2% sodium hypochlorite for one, two or three minutes, dried on a sterile absorbent paper and plated onto quarter strength Potato Dextrose Agar (Merck, Darmstadt, Germany), supple- mented with 2ml/l of 25% lactic acid to retard bacterial growth. Where fungi grew from lesions after five days, the fungus was subcultured onto half strength V8 (Campbells Soups Australia) agar and placed in a dark cabinet at 20°C until sporulation was observed. A series of limited host range tests on Avena fatua and wheat (cv. Dollarbird) were conducted for representative isolates of all genera collected. Host range testing of Drechslera avenacea The pathogenicity of twelve isolates identified as D. avenacea by the International Mycological Institute (IMI, Kew, UK) was tested against 6 cultivars of wheat, 2 cultivars of barley, 4 cultivars of oats and wild oats. The isolates chosen had geographically diverse origins. The cultivars tested were resistant to a range of diseases (Gammie 1995) (Table 1). A conidial suspension (1 x 10° conidia per ml) was made for each isolate. This was applied to 5 seedlings of all of the chosen cultivars. The seedlings were at approxi- mately the three leaf stage at the time of inoculation. Inoculated seedlings were placed in Proc. LINN. SOC. N.S.W., 119. 1998 S.D. HETHERINGTON, B.A. AAULD AND H.E.SMITH 221 TABLE | The cereal cultivars tested for their susceptibility to D. avenacea. The cultivars were chosen to represent variation in their response to a number of fungal diseases. Disease susceptiblity' Cereal Cultivar Susceptible Resistant Oats Bimbil leaf rust BYDV’ (moderately) stem rust Echidna leaf rust BYDV?° (moderately) stem rust Mortlock leaf rust BYDV? (moderately) stem rust Coolabah leaf rust BYDV’ (moderately) stem rust Barley O’ Connor net blotch (moderately) leaf scald (moderately) powdery mildew (very) covered smut Skiff leaf scald net blotch (moderately) powdery mildew (moderately) covered smut Wheat Dollarbird Septoria tritici blotch flag smut yellow spot leaf rust stem rust Rosella flag smut leaf rust stem rust Septoria tritici blotch yellow spot Hartog crown rot flag smut Septoria tritici blotch leaf rust yellow spot stem rust Janz crown rot flag smut Septoria tritici blotch (moderately) leaf rust yellow spot stem rust Sunbrook yellow spot flag smut leaf rust stem rust Septoria tritici blotch (moderately) Swift yellow spot flag smut leaf rust stem rust Septoria tritici blotch (moderately) 1. According to Gammie 1995 2. BYDV = Barley Yellow Dwarf Virus Proc. LINN. SOC. N.S.W., 119. 1998 222 POSSIBLE BIOHERBICIDE FOR AVENA FATUA a dew chamber (Percival, Iowa, USA) at 20°C for 16 hours, so that free moisture formed on the leaf surface. They were then placed in a controlled environment cabinet at 20°C with a 12 hr light/dark cycle. After ten days the percentage of necrotic tissue on the two oldest leaves was assessed visually. The average value for the five plants per treatment was used to generate an index of plant susceptibility. An isolate’s suitability as the active ingredient in a bioherbicide was based upon its fulfilment of the following criteria: 1. Index category 8 (more than 90% of leaf tissue necrotic) on wild oats 2. Index category 0, 1 or 2 (0O-4% of leaf tissue necrotic) on wheat Index category 0, 1 or 2 (0-4% of leaf tissue necrotic) on barley 3 4. Index category 5 (65% of leaf tissue necrotic) or lower on oats. RESULTS Survey of the diseases of wild oats in New South Wales One thousand two hundred and twelve fungal isolates were obtained during the survey. These isolates include Drechslera avenacea, Stagonospora avenae, Leptosphaerulina trifolii, Pleospora infectoria, Stemphylium vesicarium, Colletotrichum sublineolum, Bipolaris australiensis, Phoma subglomerata, Pyrenophora semeniperda, Epicoccum purpurascens, Curvularia sp. and Ascochyta agropyrina. Koch’s postulates were applied and confirmed that D. avenacea, S. avenae, L. tri- folii, P. infectoria, P. subglomerata and P. semeniperda infect wild oats but had little or no effect on wheat. C. sublineolum and Curvularia sp. infected both wheat and wild oats. Drechslera avenacea was chosen for closer study because of the foliar damage caused by this pathogen in artificial inoculations and its observed impact in the field. It was collected at a large number of diverse sites (Fig. 1). Some 400 isolates of this species have been lyophilised and stored. Host range testing of Drechslera avenacea The susceptibility of cultivars varied according to the identity of the challenging isolate (Table 2). No isolate fulfilled the desirable criteria perfectly. The best performing isolates were IMI374564 which caused a more severe disease reaction on all cultivars of oats than was desirable and IMI375958 which caused a more severe disease reaction on the oat cultivar Bimbil than was desirable. The majority of isolates (eight of twelve) caused severe disease on all cultivars of cultivated oats. No isolate caused severe disease on wheat or barley. DISCUSSION Genetic diversity of pathogenic fungi may be exploited in order to obtain the best isolates for use as the basis of bioherbicides. Isolates should cause severe damage to their target weed and be selective, in that they do not infect other plants; particularly valuable, closely related crop species. Widely distributed, distinct populations of fungi of the same species may demonstrate genetic variation best suiting them to particular habitats. A log- ical preliminary to examining potential bioherbicide fungi was to investigate this varia- tion through extensive surveys. While a large number of fungal species were isolated over the two year period of this study, Drechslera avenacea is the most likely to be used in a bioherbicide. The Proc. LINN. SOC. N.S.W., 119. 1998 S.D. HETHERINGTON, B.A. AAULD AND H.E.SMITH 223 GOS Sas) 0 40 80 120 160 152 Figure I. Survey sites from which isolates of Drechslera avenacea were collected during 1995 and 1996. Fungi were isolated from Avena fatua or A. ludoviciana (Q) and A. barbata (@). In other cases, (#) it was not possi- ble to determine the host identity to specific level. severity of disease it causes, its wide distribution and selectivity mean that it is potential- ly robust and effective. The isolation of fungi from infected plants in hot, dry areas (e.g. Deniliquin: 300-400 mm median annual rainfall) indicates that this isolate is either specifically adapted to these conditions or the species as a whole can tolerate a wide range of environmental conditions. Given this observation, certain isolates of this fungus may require only a short dew period; a desirable characteristic for a fungus which is to be the basis of a bioherbicide. Further testing remains to be done in this area. Should these speculations prove true the large number of isolates collected and stored during this study will enable us to further refine the bioherbicide through a more intensive study of other isolates. Isolates have also been collected from southern Queensland, Victoria and Western Australia. The susceptibility of cultivated oats to isolates tested is a concern. The fungus caus- es eye spots, pre-emergence and post-emergence seedling blight, leaf stripes of young plants and leaf spots of mature oats (Sivanesan 1987); crop losses of 2—10% have been reported in Finland (Rekola et al. 1970). However, two points should be considered. Firstly, this study identified a great deal of variation in the virulence of isolates; for exam- ple, for the cultivar Echidna, the area of necrotic leaf tissue as a result of inoculation var- ied from 7% to 97.2% dependent upon which isolate was applied. Careful screening of Proc. LINN. Soc. N.S.W., 119. 1998 224 POSSIBLE BIOHERBICIDE FOR AVENA FATUA TABLE 2 Susceptibility 1 of a range of cereal cultivars to twelve isolates of Drechslera avenacea which were collected from regions throughout New South Wales. Highlighted numbers indicate interactions which fulfilled the crite- ria desirable for the isolate to be used as a bioherbicide outlined in the text. Isolate accession number ~* 900066600 6 © 6 MI375692 ~~» @0006606060 0 6 © 37591 ~~ @9O0G0O00000O™ 375959 Cultivar Bimbil Echidna Mortlock Coolabah O’ Connor Skiff Dollarbird Rosella Hartog Janz Sunbrook Swift Wild Oats OOOO OOO68 = = = & M374568 0» 0000060 © = = & ™374565 000-060 +06006=& & &~ &~ IM375957 ~» @00+6006060 = °o & &~ IMB374572 ~ @OOOOO06 = = = o M374567 a» O©O» OO» COO = ~ & IM374566 ©- 60» Ww ff Ww wo & ~ ec 600 «600s s TMI374570 ©000000060006 = 1m375958 O9OOO00060~ ~ = = MB74564 5 6 6 ' 0: 0% of leaf area necrotic; 1: 0-2% of leaf area necrotic; 2: 24% of leaf area necrotic; 3: 4-9% of leaf area necrotic; 4: 9-50% of leaf area necrotic; 5: 50-65% of leaf area necrotic; 6: 65-80% of leaf area necrotic; 7: 80-90% of leaf area necrotic; 8: 90-100% of leaf area necrotic; x: interaction not assessed. isolates will maximise selectivity. Secondly, while dispersal mechanisms for D. avenacea have not been studied, it may be assumed that they are typical of the “Helminthosporium” group in which conidia are dispersed only a short distance by rain splash, wind-driven rain, or air currents. Because spores are relatively large, dispersal is only over short dis- tances (Agrios 1997). Applications of D. avenacea as a bioherbicide in wheat cultivation are unlikely to spread to nearby oat cultivation. Through the protocol adopted in this study a number of isolates have been selected for further study. Studies of disease etiology remain to be done. ACKNOWLEDGEMENT This work was undertaken with the financial support of the Australian Centre for International Agricultural Research (ACIAR). REFERENCES Agrios, G.N. (1997). “Plant Pathology’. 4th edition. (Academic Press Inc., San Diego, USA). Evans, H.C. (1991). Biological control of tropical grassy weeds. In ‘Tropical Grassy Weeds’ (Eds F.W.G. Baker and P.J. Terry). pp. 52-72. 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Graminicolous species of Bipolaris, Curvularia, Drechslera, Exserohilum and their teleomorphs. CAB International. Mycological papers No. 158. pp. 162—163. Whalley, R.D.B. and Burfitt, J.M. (1972). Ecotypic variation in Avena fatua L., A. sterilis L. (A. ludoviciana), and A. barbata Pott. in New South Wales and southern Queensland. Australian Journal of Agricultural Research 23, 799-810. Wilson, S. (1987). The scope for biological control of Avena fatua L. with Drechslera avenae (Eidam) Sharif. PhD thesis, Oxford University, Oxford. Zonjian, Z. and Yanghan, L. (1996). Discovery, isolation and pathogenicity study of a wild oat (Avena fatua) biological control fungus. Proceedings of the III International Bioherbicide Workshop. Stellenbosch, South Africa. p. 30. Proc. LINN. Soc. N.S.W., 119. 1998 Hse, nse witest i a Ni Een» Barta icone) learns Mnige: iba at: “yi ‘ } ae {BRITE Sy gine’ aye ie: 5h A a. al me et ; PENMESE SR eoTTy Res diate nk MBG > Nee fies epee e. 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