amphibian-reptile-conservation.org Published in the United States of America 2013 • VOLUME 6 • NUMBER 4 AMPHIBIAN & REPTILE ISSN: 1083-446X elSSN: 1525-9153 Editor Raul E. Diaz University of Kansas, USA Craig Hassapakis Berkeley, California, USA Associate Editors Howard O. Clark, Jr. Erik R. Wild Garcia and Associates, USA University of Wisconsin-Stevens Point, USA Assistant Editors Alison R. Davis University of California, Berkeley, USA Daniel D. Fogell Southeastern Community College, USA Editorial Review Board David C. Blackburn California Academy of Sciences, USA Bill Branch Port Elizabeth Museum, SOUTH AFRICA Jelka Crnobrnja-Isailovc IBISS University of Belgrade, SERBIA C. Kenneth Dodd, Jr. Lee A. Fitzgerald Adel A. Ibrahim University of Florida, USA Texas A&M University, USA Ha’il University, SAUDIA ARABIA Harvey B. Lillywhite Julian C. Lee Rafaqat Masroor University of Florida, USA Taos, New Mexico, USA Pakistan Museum of Natural History, PAKISTAN Peter V. 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Wilson AMNH Southwestern Research Station, USA Carl C. Gans (1923-2009) Honorary Members Joseph T. Collins (1939-2012) Cover : Neurergus kaiseri. Amphibian & Reptile Conservation — Worldwide Community-Supported Herpetological Conservation (ISSN: 1083-446X; elSSN: 1525-9153) is published by Craig Hassapakis /Amphibian & Reptile Conservation as full issues at least twice yearly (semi-annually or more often depending on needs) and papers are immediately released as they are finished on our website; http://amphibian-reptile-conservation.org; email: arc.publisher@gmail.com Amphibian & Reptile Conservation is published as an open access journal. Please visit the official journal website at: http://amphibian-reptile-conservation.org Instructions to Authors : Amphibian & Reptile Conservation accepts manuscripts on the biology of amphibians and reptiles, with emphasis on conservation, sustainable management, and biodiversity. Topics in these areas can include: taxonomy and phytogeny, species inventories, distri- bution, conservation, species profiles, ecology, natural history, sustainable management, conservation breeding, citizen science, social network- ing, and any other topic that lends to the conservation of amphibians and reptiles worldwide. Prior consultation with editors is suggested and important if you have any questions and/or concerns about submissions. Further details on the submission of a manuscript can best be obtained by consulting a current published paper from the journal and/or by accessing Instructions for Authors at the Amphibian and Reptile Conservation website: http://amphibian-reptile-conservation.org/submissions.html © Craig Hassapakis! Amphibian & Reptile Conservation Copyright: © 2012 Sharifi et al. This is an open-access article distributed under the terms of the Creative Com- mons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Amphibian & Reptile Conservation 6(4): 1-8. Sexual size dimorphism in Neurergus kaiseri (Caudata: Salamandridae) in south-western Zagros Mountains, Iran ^ozafar Sharifi, Hossein Farasat, and Somaye Vaissi Rctzi University Center for Environmental Studies , Department of Biolog}’, Faculty of Science, Baghabrisham 67149, Kermanshah, IRAN Abstract . — Using bivariate and multivariate techniques we evaluated sexual size dimorphism in 13 body-related and six head-related metrics from 99 live specimens of the Lorestan newt, Neurergus kaiseri. Analyses of variance of 12 metrics showed that average sizes for all these characters in fe- males are significantly (P< 0.05) greater than in males. However, one character (vent length) is larger in males than females (P<0.001). Evaluation of 13 metrics showed that average size dimorphism is apparent in 10 characters with nine characters showing these differences at P<0.01 and one char- acter at P<0.05 confidence levels. Principal Components Analysis of external characters provided a good separation of males and females. Although body measurements gave a clear pattern of differ- ences between the sexes in N. kaiseri, head measurements showed no such distinctions. Key words. Neurergus kaiseri , Principal Component Analysis (PC A), sexual size dimorphism, southwestern Iran, cloaca Citation: Sharifi M, Farasat H, Vaissi S. 2012. Sexual size dimorphism in Neurergus kaiseri (Caudata: Salamandridae) in south-western Zagros Moun- tains, Iran. Amphibian & Reptile Conservation 6(4):1-8(e48). Introduction Sexual dimorphism shows widespread and recogniz- able patterns in many species and has been studied for more than a century (reviewed in Fairbairn et al. 2007). Sexual size dimorphism is common in animal taxa, but is highly variable in magnitude and direction (Anders- son 1994; Fairbairn 1997; Brandt and Andrade 2007). Sexually dimorphic traits have been surveyed in differ- ent classes of vertebrates, including birds (Temeles 1985; Temeles et al. 2000), primates (Crook 1972), amphibians (Schauble 2004; Vargas-Salinas 2006; McGarrity and Johnson 2008; Malmgren and Thollesson 1999; Kalezic et al. 1992), lizards (Bruner et al. 2005; Kaliontzopou- lou et al. 2007), and snakes (Feriche et al. 1993; Shine et al. 1999). Amphibian females generally grow larger than males and female body size is often correlated to clutch size (Duellman and Trueb 1986; Rafinski and Pecio 1989; Kalezic et al. 1992). In amphibians, the most strikingly dimorphic sexual characteristics are seasonal; however, most species also show permanent sexual dif- ferences in morphometries and morphology (Malacarne and Cortassa 1983). Although mature female amphibians are generally larger than males, and female body size is often corre- lated to clutch size, there are examples where males are the larger. This can be attributed to high degrees of ago- nistic male behavior such as combat during the reproduc- Correspondence. Email: 1 sharifimozafar2012@gmail.com tive season. Several theories have been developed to ex- plain ecological and evolutionary significance for sexual size dimorphism (e.g., Slatkin 1984; Andersson 1994). As stated by Daiwin (1871) sexual selection is likely the most important single cause that generates dimorphism, but other factors such as female reproductive strategy and competition for food resources have been considered to be significant (Duellman and Trueb 1986). In the present paper, we explore and discuss sexual size dimorphism in the Lorestan newt Neurergus kai- seri in the southwestern mountains of Iran. The aim is to describe the expression of intersexual differences in this species to reveal sexually dimorphic traits that can be important in systematic and evolutionary research. Material and methods We measured 99 live specimens of Neurergus kaiseri found in the southern Zagros ranges. The average annual precipitation in the southern Zagros ranges from 400 to 800 mm per year. The dominant vegetation cover around streams is oak tree ( Quercus brantii ) open woodlands. The active period of N. kaiseri in its aquatic environment starts in March and ends in July, a period when tempera- ture enables breeding and feeding. The N. kaiseri used in the present study (58 males, 41 females) were all caught in the daytime between the 7 th and 13 th April 2012. The amphibian-reptile-conservation.org 001 July 201 2 | Volume 6 | Number 4 | e48 Sharifi et al. Figure 1 . Male Neurergus kaiseri have a fleshy protuberance at the base of the tail (A), whereas female has a prominent cloaca but without the protuberance (B). Photos by Mozafar Sharifi. method of capture was by hand, with individuals taken from among or under stones in the shallow water at the side of the stream. After measuring, the N. kaiseri were released unhanned at the location of their capture. The sex of each individual was assessed according to external secondary sexual characters: males have a fleshy protu- berance at the base of the tail, whereas females have a prominent cloaca, but without the protuberance (Fig 1). Juveniles are differentiated from mature adults in having a smaller body length and lacking both the protuberance and the prominent cloaca (Baran and Atatiir 1998). Thirteen variables (Table 1) for all specimens were measured with calipers to the nearest 0.1 mm. To test significant differences of sexually dimorphic metrics, Independent Sample /-test (2 -tailed) as well as Principal Component Analysis (PCA: correlation matrix) at the significance level of 0.01 were used. In multivariate anal- yses the variables were carried out into two sets (body- and head-related measurements) and were analyzed separately using PCA based on Pearson’s correlation coefficients. SPSS software version 16, Excel, and Past software were used for running the statistical analyses. Table 1 . Definitions of the morphometric character set and abbreviations used for body- and head-related metrics. Abbreviations Body measurements W SVL TL LFL LHL FHL VL TLL Head measurements HL HW HH IOD DN LW Variable definition Weight Snout to vent length, tip of snout to anterior margin of cloacal lips Tail length, posterior margin of cloacal lips to tip of tail Length of fore limb, anterior margin of front leg to tip of the longest finger Length of hind limb, anterior margin of hind leg to tip of the longest toe Forelimb to hindlimb length, posterior margin of front leg (axilla) to anterior margin of hind leg (groin) Vent length, anterior margin of cloacal lips to posterior margin of cloacal lips Total length, tip of snout to tip of tail (SVL + VL + TL) Head length, tip of snout to posterior region of neck Head width, largest width of head, in line with the comer of the mouth Height of head, margin of lower jaw to upper of eye, in line with the eyes Interorbital distance, shortest distance between eyes Distance of nostrils, from one nostril to the other Length of wrinkles under throat, tip of snout to posterior margin of wrinkles under throat amphibian-reptile-conservation.org 002 July 2012 | Volume 6 | Number 4 | e48 Sexual size dimorphism in Neurergus kaiseri Results The results of the Independent Sample t-test (2 -tailed) show that in N. kaiseri most characters differed signifi- cantly between sexes (P<0.01), so that in each variable females are larger than males, excluding vent length (Ta- ble 2). In N. kaiseri, 89% of body related metrics were significantly sexually dimorphic (Table 2). In contrast, there were fewer such sex related differences (50%) in measurements related to head morphology. The mean male to female Snout to Vent Length (SVL) ratio was 0.86 for N. kaiseri (Table 2). In Bivariate Analyses, SVL in relation to the Total Length (TL; measured from tip-of-snout to tip-of-tail), was dimorphic in N. kaiseri (males 46.59% ± 0.004 SE, females 49.36% ± 0.004 SE; P <0.001). The relation between Tail Length (TL) to the Total Length and Lorelimb to Hindlimb Length (LHL) to SVL were not significantly sexually dimorphic in N. kai- seri. Males of N. kaiseri had the shorter tail (males 56.76 ± 0.93 SE, females 63.20 ± 0.92; P <0.001). Analysis of metrics indicated general trends in varia- tion. Loadings for the first two components are given in Table 2, and the individual specimens are projected onto these components in Ligure 3. In each PC A there were high positive loadings for all characters on PCI. This axis is therefore interpreted as a general size measure. Contrasting positive and negative loadings were found on PC2, indicating general shape measures as important for this separation. Body variables gave a clear pattern of differences be- tween the sexes in N. kaiseri, while head measurements showed no such distinctions (Pigs. 1 and 2). Lactor load- ings for principal components (Table 3) revealed that a total of 75.9% and 58.3% of the variability for N. kaiseri could be explained by the first two components (PCI and PC2) for body- and head-related traits, respectively. The first component, which explained 62.1% and the second component, which explained 13.7% of the total character variation for body characters, provided complete separa- tion between males and females (Pigs. 3, 4, and 5). The first component, which explained 39% and the second component, which explained 19.3% of the total character variation for head measurements, do not prove complete separation between males and females (Pigs. 3, 4, and 5). The remaining components (PC3 ± PC9) individually explained < 12% of the total variation for this species on body related traits, and did not reveal any readily inter- pretable patterns. Lactor loadings for discriminant Hot- teling’s T 2 revealed that with using body measurements, males and females were well separated (Hotteling’s T 2 : 1307.9, P: 175.28, PO.OOl), but with using head variables the sexes were not separated (Hotteling’s T 2 : 29.351, P: 5.63, P0.001), and revealed these measure- ments unsuitable for determination of sexual dimorphism in N. kaiseri ( Pig. 2). Table 2. Descriptive statistics (mean, standard error of mean, and range) of 13 external characteristics (mm) in males and females of Neurergus kaiseri ; n: number; SE: standard error of mean. Morphometric Abbreviations: W (Weight), SVL (Snout- Vent Length), TL (Tail Length), HH (Height of Head), LFL (Length of Forelimb), LHL (Length of Hindlimb), FHL (Forelimb to Hindlimb Length), VL (Cloacal Length), IOD (Interorbital Distance), DN (Distance of Nostrils). Variable Males (n = Mean ± SE 58) Range Females ( n Mean ± SE = 41) Range P Body measurements W 6.2 ± 0.2 3 . 2 - 9 . 8 7.2 ± 0.3 3 . 8 - 11.4 0.0 SVL 54.8 ± 0.5 47 . 9 - 61.8 63.6 ± 0.8 54.1 - 78.9 0.0 TL 56.8 ± 0.9 33 . 5 - 72.8 63.3 ± 0.9 52 . 6 - 75.9 0.0 LFL 20.1 ± 0.2 16 . 3 - 22.3 20.5 ± 0.2 16 . 9 - 23.1 0.05 LHL 21.6 ± 0.2 17 . 2 - 24.6 22.4 ± 0.3 19 . 2 - 26.0 0.0 FHL 29.4 ± 0.4 18 . 0 - 35.8 35.0 ± 0.6 26 . 9 - 41.6 0.0 LW 13.8 ± 0.2 11.1 - 16.7 14.4 ± 0.4 6 . 2 - 18.9 0.1 VL 6.2 ± 0.2 4 . 5 - 7.9 2.1 ± 0.4 1 . 4 - 2.5 0.0 TLL 117.8 ± 1.4 100.1 - 137.3 128.9 ± 1.4 111 . 8 - 146.5 0.0 Head measurements HL 13.5 ± 0.1 10 . 7 - 16.0 13.6 ± 0.1 12.1 - 15.0 0.2 HW U . 0 ± 0.1 9 . 5 - 13.4 11.1 ± 0.2 5 . 7 - 12.5 0.7 HH 5.6 ± 0.1 4 . 9 - 7.4 6.0 ± 0.1 5 . 3 - 7.2 0.0 IOD 7.1 ± 0.1 L/< to 1 oo o 7.4 ± 0.1 6 . 2 - 8.8 0.0 DN 3.8 ± 0.1 1 . 2 - 4.6 4.0 ± 0.0 3 . 5 - 4.7 0.01 amphibian-reptile-conservation.org 003 July 201 2 | Volume 6 | Number 4 | e48 Sharifi et al. Figure 2. Discriminant Hotelling’s T 2 for separation of male and female Neurergus kaiseri. (A) Body-related characters; (B) Head- related variables. Note that on base head-related variables male and female aren’t well separated. Figure 3. Principal Component Analysis (PCA) on sexual dimorphism in Neurergus kaiseri. Scatter plots of principal component scores for the first two principal axes, with convex polygons for males and females. Loadings are shown in Table 3. (A) Body- related characters; (B) Head-related variables, significant sex differences are noted on neither PCI, nor PC2. PCI ( 55.82 %) Figure 4. Principal Component Analysis (PCA) on seasonal sexual dimorphism in Neurergus kaiseri. Scatter plots of prin- cipal component scores for the first two principal axes, with convex polygons for males and females. Loadings are shown in Table 3. Discussion The Lorestan newt Neurergus kaiseri was shown not to exhibit sexually dimorphism in head-related metrics. This is in agreement with other studies on head morphol- ogy in newts (Malmgren and Thollesson 1999; Rafinski and Pecio 1989; Kalezic et al. 1992). These results do not support the ecological model that N. kaiseri has de- veloped intersexual differences in feeding strategies along a niche divergence process (Slatkin 1984; Ander- sson 1994) driven by the two factors, the rate of feed- ing and type of food consumed (Shine 1989). The first factor considers that substantial intersexual difference in body size lead to differences in feeding rates between the sexes. The second factor comprises species where the sexes diverge in trophic morphology as a result of inter- sexual differences in dietary preferences. Both male and female newts experience high energetic costs during the reproductive season (Halliday and Arano 1991; Griffiths amphibian-reptile-conservation.org 004 July 201 2 | Volume 6 | Number 4 | e48 Sexual size dimorphism in Neurergus kaiseri SVL 7.50 7.00 6.50 ^ 6.00 5.50 5.00 4.50 Figure 5. Bivariate scatterplots of variables on sexual dimorphism with convex polygons for males and females in Neurergus kai- seri. (A and B) Body-related characters (C and D) Head-related variables. Body measurements gave a clear pattern of differences between the sexes in N. kaiseri , while head measurements showed no such distinctions. All values in mm. 1996) and both spend considerable time feeding when not involved in courtship. This indicates that feeding rates between males and females might be similar within species, even during the breeding period, suggesting that sexual dimorphism resulting from feeding rates and diet may be negligible. The separation of sexes in statistical analyses was high in N. kaiseri. Sexual dimorphism was attributed to females showing large values for dimensions related to fecundity, such as SVL and distance of FHL, contrasted with large values for cloaca in males. In all analyses the female SVL and FHL metrics were highly significant in the observed patterns, contrasting against the male CL. These results can be interpreted as primarily concordant with the fecundity model. Previous studies on amphib- ians have shown that females are generally larger than males in body size (Duellman and Trueb 1986), possibly because fecundity increases with increasing female body size. Males, however, can often increase their lifetime re- productive success through other life history traits in spe- cies with little or no agonistic behavior; for example by maturing at an early age. Moreover, Kalezic et al. (1992), showed that the trunk length (corresponding to FHL) is directly correlated to the length of the pleuroperitoneal cavity in Triturus newts to which Neurergus is a closely related. amphibian-reptile-conservation.org 005 July 201 2 | Volume 6 | Number 4 | e48 Sharifi et al. Table 3. Factor loadings for the first two principal components (eigenvectors) for Neurergus kaiseri from multivariate analyses (Principal Components Analysis, PC A) on body- and head-related variables. Neurergus kaiseri Variable PCI PC2 PC3 Body measurements SVL 0.848 0.124 - 0.304 TL 0.755 - 0.570 0.293 LFL 0.668 0.470 0.447 LHL 0.734 0.420 0.041 FHL 0.762 - 0.025 - 0.534 TLL 0.934 - 0.298 0.123 Eigenvalue 3.726 0.827 0.680 % of variability 62.099 13.776 11.328 Cumulated % 62.099 75.874 87.202 Head measurements HH 0.484 0.794 0.049 DN 0.631 - 0.540 - 0.003 IOD 0.654 0.140 - 0.482 Eigenvalue 1.699 0.985 0.887 % of variability 39.011 19.328 17.216 Cumulated % 39.011 58.339 75.555 Male N. kaiseri could be distinguished from females in having a larger cloaca (Fig. 1). The cloacal swelling in male newts is most notable laterally and ventrally com- pared to females, and may be an important factor in male mating success. Most of the cloacal volume is occupied by glands secreting substances fonning the spermato- phore, although tubules emanating from the pheromone- producing dorsal gland are present — especially in the caudal region of the cloaca (Sever et al. 1990). The dor- sal gland itself, which is known to be greatly enlarged during the breeding season in some newts, lies anterior to the pelvic girdle. As proposed by Sever et al. (1990), it is quite likely that both the rate of spennatophore pro- duction and the synthesis of courtship pheromones — fac- tors contributing to male mating success — are under the influence of sexual selection, thus increasing the size and volume of structures in the cloacal region. Acknowledgments. — We thank Nate Nelson for pro- viding funding for this project through the conservation breeding program for N. kaiseri at Sedgewick County Zoo, Wichita, Kansas, USA. We are also grateful to Razi University which provides funding to the postgraduate students involved in present study. Literature cited Andersson M. 1994. Sexual selection. Princeton Univer- sity Press, Princeton, New Jersey, USA. 1 Figure 6. Factor loadings for the first two principal components in analyses of body-related characters. Loadings on PCI are all close to one and positive, and this component is interpreted as a general size measure. On PC2 positive loadings (characters above the abscissa) are contrasted with negative loadings (be- low), and the component is interpreted as a measure of shape that discriminates between males and females (Figs. 4 and 5). Abbreviations as in Table 1 . amphibian-reptile-conservation.org 006 July 2012 | Volume 6 | Number 4 | e48 Sexual size dimorphism in Neurergus kaiseri Baran I, Atatiir MK. 1998. Turkish Herpetofciuna (Am- phibians and Reptiles). Republic of Turkish Ministry of Environment, Ankara, Turkey. Brandt Y, Andrade MCB. 2007. Testing the gravity hy- pothesis of sexual size dimorphism: Are small males faster climbers? Functional Ecology 21(2):379-385. Bruner E, Costantini D, Fanfani A, DelTomo G. 2005. Morphological variation and sexual dimorphism of the cephalic scales in Lacerta bilineata. Acta Zoolog- ica (Stockholm) 86(4):245-254. Crook JH. 1972. Sexual selection, dimorphism, and so- cial organization in the primates. In: Sexual Selection and the Descent of Man. Editor, Campbell B. Aldine Publishing Company, Chicago, Illinois, USA. 231- 281. Darwin C. 1871. The Descent of Man, and Selection in Relation to Sex. Murray, London, United Kingdom. Duellman WE, Trueb L. 1986. Biology of Amphibians. McGraw-Hill, New York, New York, USA. Fairbaim DJ, Blanckenhom WU, Szekely T. (Editors). 2007. Sex, Size and Gender Roles: Evolutionary Stud- ies of Sexual Dimorphism. Oxford University Press, Oxford, United Kingdom. Fairbaim DJ. 1997. Allometry for sexual size dimor- phism: Pattern and process in the coevolution of body size in males and females. Annual Reviews of Ecology and Systematics 28:659-687. Feriche M, Pleguezuelos JM, Cerro A. 1993. Sexual dimorphism and sexing of Mediterranean colubrids based on external characteristics. Journal of Herpe- tologv 27(4):357-362. Griffiths RA. 1996. Newts and Salamanders of Europe. Academic Press, London, United Kingdom. Halliday T, Arano B. 1991. Resolving the phylogeny of the European newts. Trends in Ecology and Evolution 6(4): 11 3- 117. Kalezic ML, Cmobrnja J, Dorovic A, Dzukic G. 1992. Sexual size difference in Triturus newts: Geographical variation in Yugoslav populations. Alytes 10(2):63-80. Kaliontzopoulou A, Carretero MA, Llorente GA. 2007. Multivariate and geometric morphometries in the analysis of sexual dimorphism variation in Podarcis lizards. Journal of Morphology 268(2): 152-165. Malacarne G, Cortassa R. 1983. Sexual selection in the crested newt. Animal Behaviour 3 1(4): 1256-1257. Malmgren JC, Thollesson M. 1999. Sexual size and shape dimorphism in two species of newts, Triturus cristatus and T. vulgaris (Caudata: Salamandridae). Journal of Zoology 249(2): 127-136. Mcgarrity ME, Johnson SA. 2008. Geographic trend in sexual size dimorphism and body size of Osteopilus septentrionalis (Cuban treefrog): Implications for in- vasion of the southeastern United States. Biological Invasions 11 (6): 141 1-1420. Rafinski J, Pecio A. 1989. Craniometric studies on the species of the genus Triturus, Rafmesque, 1815 (Am- phibia, Salamandridae). Folia Biologica (Krakow) 37:131-150. Schauble CS. 2004. Variation in body size and sexual dimorphism across geographical and environmental space in the frogs Limnodynastes tasmaniensis and L. peronii. Biological Journal of the Linnean Society (London) 82(l):39-56. Sever DM, Verrell PA, Halliday TR, Griffiths M, Waights V. 1990. The cloaca and cloacal glands of the male smooth newt, Triturus vulgaris vulgaris (Linnaeus), with special emphasis on the dorsal gland. Herpeto- logica 46(2): 160-168. Shine R, Olsson MM, Moore IT, Lemaster MP, Mason RT. 1999. Why do male snakes have longer tails than females? Proceedings of the Royal Society of London B (Biological Sciences) 266(1434):2147-2151. Shine R. 1989. Ecological causes for the evolution of sexual dimorphism: A review of the evidence. Quar- terly Review of Biology 64(4):419-461. Slatkin M. 1984. Ecological causes of sexual dimor- phism. Evolution 38(3): 135-139. Temeles EJ, Pan IL, Brennan JL, Horwitt JN. 2000. Evi- dence for ecological causation of sexual dimorphism in a hummingbird. Science 289(5478):441-443. Temeles EJ. 1985. Sexual size dimorphism of bird-eating hawks: The effect of prey vulnerability. American Naturalist 125(4):485-499. Vargas-Salinas F. 2006. Sexual size dimorphism in the Cuban treefrog Osteopilus septentrionalis . Amphibia- Reptilia 27(3):419-426. Received: 11 May 2012 Accepted: 07 June 2012 Published: 12 July 2012 amphibian-reptile-conservation.org 007 July 201 2 | Volume 6 | Number 4 | e48 Sharifi et al. Mozafar Sharifi is a senior lecturer in ecology at Department of Biology, Razi University, Kerman- shah, Iran. He is also director of Razi University Center for Environmental Studies. In recent years his main research interest focuses on conservation biology of chiroptera and amphibians. He has contribu- tion to the processes involving conservation assessment of chiroptera and two species the genus Neu- rergus in collaboration with the IUCN. Hossain Farasat is currently a Ph.D. candidate at Department of Biology, Razi University, Kerman- shah, Iran. He earned his M.Sc. from Razi University. His present research focuses on the ecology and genetic diversity of fragmented populations of Neurergus kaiseri. His main interest is to examine whether these fragmented populations are structured by a metapopulation. He is also keen to apply his finding in conservation of this critically endangered and endemic species of Iran. Somaye Vaissi is a M.Sc. student in systematic zoology at Department of Biology, Razi University, Iran. She has earned her B.Sc. in animal biology from the same department. She is currently the curator of a Captive Breeding Facility for Neurergus microspilotus at Razi University funded by the Mohamed bin Zayed Species Conservation Fund. Her current research activities with two species of Neurergus involve several topics associated with husbandry and health of the newts in captivity. These include nutrition, growth, development and their health. She has contribution in detecting chytrid fimgus and other diseases such as red-leg syndrome and rickettsial inclusions in the newts. amphibian-reptile-conservation.org 008 July 2012 | Volume 6 | Number 4 | e48 Copyright: © 2012 Bogaerts et al. This is an open-access article distributed under the terms of the Creative Com- mons Attribution-NonCommercial-NoDerivs 3.0 Unported License, which permits unrestricted use for non-com- mercial and education purposes only provided the original author and source are credited. Amphibian & Reptile Conservation 6(4):9-29. Conservation biology, husbandry, and captive breeding of the endemic Anatolia newt, Neurergus strauchii Steindachner (1887) (Amphibia: Caudata: Salamandridae) ^erge Bogaerts, 2 Henry Janssen, Jennifer Macke, 4 Gunter Schultschik, 5 Kristina Ernst, 6 Frangois Maillet, 7 Christoph Bork, 8 Frank Pasmans, and Patrick Wisniewski Hupinelaan 25, NL-5582CG Waalre, THE NETHERLANDS 2 Calvariebergstraat 6, B-8000 Brugge, BELGIUM 3 675 Totavi Street, Los Alamos, New Mexico 87544, USA 4 Sachsenweg 6, Haus 12, A-2391 Kaltenleutgeben, AUSTRIA 5 Waldgartenstrasse 26, D-81377 Miinchen, GERMANY *24 Rue du Bondar, F-95740 Frepillon, FRANCE D-44359 Dortmund, GERMANY ^Laboratory of Veterinary’ Bacteriology and Mycology Faculty of Veterinary Medicine, Ghent University ’, Salisburylaan 133, B-9820, Merelbelce, BELGIUM Abstract. — The long-term experiences of different private breeders on husbandry and breeding of the Anatolia newt, Neurergus strauchii are presented. This information is introduced and discussed in respect to the ecology, systematics, and conservation of N. strauchii. Our knowledge and data of husbandry and captive breeding is collated and compared with the literature. We present our experi- ences to provide information and advice for the successful long-term keeping, breeding, and raising of N. strauchii and also an example and model that may be used for privates’ contribution to Conser- vation Breeding Programs for endangered Neurergus species and other semi-aquatic salamanders. Neurergus strauchii has proved relatively easy to keep in captivity under a range of aquatic and terrestrial housing and with adequate diet. However, although breeding is successful under a vari- ety of conditions survival from egg to adult is low. Cold husbandry temperatures in winter increase reproduction. Eggs are laid very irregularly in time and number, and oviposition may depend on the condition of the female, particularly her nutritional condition through diet. There may be up to 285 eggs per female. The best temperature for egg laying is about 14.5 °C. Hatching success of eggs can vary enormously from 0% to 80%. Most larvae hatch from 11.5 to 14.5 mm. Larvae are easy to raise, with low mortality over a wide range of temperatures, and metamorphose in three to seven months, mostly from 55 to 63 mm and about 0.6 g. Several diseases are known to affect these newts and high temperature stress may exacerbate pathology. Key words. Neurergus strauchii, breeding, husbandry, ecology, conservation, private breeders, long-term mainte- nance, diseases, international cooperation Citation: Bogaerts S, Janssen H, Macke J, Schultschik G, Ernst K, Maillet F, Bork C, Pasmans F, Wisniewski P. 2012. Conservation biology, husbandry, and captive breeding of the endemic Anatolia newt, Neurergus strauchii Steindachner (1887) (Amphibia: Caudata: Salamandridae). Amphibian & Rep- tile Conservation 6(4):9-29(e53). Introduction Since its description by Steindachner (1887), relatively little information has been collected on the Anatolia newt, Neurergus strauchii. Schmidtler and Schmidtler (1970) were the first to collect substantial information on this species. In 1982, the first captive breeding expe- riences were published by Fleck (1982). Haker (1985) described breeding an F2 generation and the appearance of a color mutant, later known as the “gold-dust” variety. Although Fleck and Haker both mentioned that it was not difficult to keep and breed N. strauchii, it is still rela- tively rare to find N. strauchii in captivity. Little informa- tion on the husbandry of N. strauchii has been published, perhaps due to a lack of husbandry and breeding success. Steinfartz (1995) was the first to report detailed informa- tion on the keeping and breeding of the subspecies N. s. barani, which had been described just two years prior (Oz 1994). Inspired by the aquatic versus terrestrial rearing ex- periments on juvenile N. s. strauchii of Jennifer Macke (Macke 2006), the scattered Internet data sheets (see for instance Schultschik 2010; Sparreboom 2009), and the fact that Kristina Ernst is running a Studbook for this species for the AG Urodela, Serge Bogaerts started col- lecting data and experiences from active and long-term breeders in order to establish some guidelines for suc- cessful husbandry of this species. In 2007, our common project was presented at the meeting of the Arbeitsgruppe Correspondence. Email: 2 hemy.janssen@skynet.be, fpmacke@gmail. com, Hnfo@salamanderland.at, -tina.ernst@yahoo.de, f ug.maillet@free.fi: 1 christoph.bork@t-online.de, frank.pasmans@ugent.be, 1 s-bogaerts@hetnet.nl (corresponding author). 1 We dedicate this paper to a passionate and experienced amphibian keeper and breeder, Patrick Wisniewski, who sadly passed away during the time of writing. amphibian-reptile-conservation.org 09 September 2012 | Volume 6 | Number 4 | e53 Bogaerts et al. Figure 1. Captive bred adult female of Neurergus s. strauchii. Photo by Serge Bogaerts. Figure 2. Adult female of Neurergus strauchii barani photographed at Kubbe mountain, Malatya. Photo by Serge Bogaerts. Urodela of the Deutsche Gesellschaft fur Herpetologie und Terrarieukunde (DGHT) in Gersfeld, Germany (Bo- gaerts 2007). Not all authors have collected similar data for example, Henry Janssen has put an extraordinary ef- fort in collecting data on reproduction between 1991 and 1997. However, through collating all husbandry knowl- edge and data, we can draw some general guidelines for successfully keeping and breeding N. strauchii. We will combine the infonnation from both subspecies, as there appears to be few differences in their maintenance. Distribution, description, and habitat Neurergus strauchii is endemic to mountainous areas in eastern Turkey, roughly from Malatya to Lake Van. The subspecies N. s. barani is found only in the mountains southeast of Malatya. Neurergus s. strauchii has a wider distribution and is found east from the river Euphrates amphibian-reptile-conservation.org up onto the Lake Van area. Although there is a relatively high level of genetic differentiation at both the mitochon- drial (12S and 16S rRNA) and nuclear levels between the subspecies (Steinfartz et al. 2002; Pasmans et al. 2006), it is not very easy to distinguish individuals of each sub- species, particularly as juveniles. Ozdemir et al. (2009) found that N. s. barani is not strongly differentiated from N. s. strauchii, suggesting their distributions are either connected, or have been separated only recently. The most obvious visual difference between N. s. strauchii and N. s. barani is the difference in the num- ber and size of yellow spots on adults (Figs. 1 and 2). The main phenotypic difference between the subspecies is that the number of spots greatly increases during matu- ration in N. s. strauchii, but increases veiy little in N. s. barani. The N. s. barani subspecies keeps approximately its juvenile pattern of small spots in two rows dorsally, September 2012 | Volume 6 | Number 4 | e53 010 Conservation, husbandry, and captive breeding of the Anatolia newt, Neurergus strauchii Figure 3. Cloaca’s of male (left) and female (right) of N. s. bamni during breeding. Photo by Serge Bogaerts. whereas the number of spots on N. s. strauchii increases as it matures. Although this difference is very pronounced between the eastern populations of N. s. strauchii and N. s. barani, the westernmost N. s. strauchii are virtually in- distinguishable from N. s. barani in this respect. Pasmans et al. (2006) found a geographically correlated increase in the number of spots on adult newts towards the eastern part of their distribution. Neurergus strauchii are relatively large newts, mea- suring up to 19 cm (Steindachner 1887). Mean lengths in the wild are 14.3 cm for adult males and 15.2 cm for adult females (Table 1). The largest total length docu- mented in the field was 18.1 cm for a female (n = 42) and 17.6 cm for a male (n = 21) (Pasmans et al. 2006). Males can be recognized by their slender body, shorter tale, larger cloaca, and the bluish-white colorations on the lateral sides of the tail, which can run through to the lateral sides of the body. These breeding colorations are often already visible in autumn. Females have an orange cloaca, relatively longer tails, and shorter legs, and ap- pear more robust than males (Fig. 3). Neurergus strauchii lives roughly between 1,000 and 1 ,900 m. above sea level. Its breeding habitats are moun- tain brooks, preferably with large, deep, slow running pools. A typical habitat is shown in Figure 4. Terrestrial habitats are often very bare, without much vegetation (Bo- gaerts et al. 2006). Water temperatures vary considerably seasonally and with stream length from springs. Pasmans et al. (2006) recorded water temperatures in breeding streams from 10.9 to 17.3 °C, although Schmidtler and Schmidtler (1970) recorded temperatures of 9 to 10 °C in a flowing spring in which they found adults. Schnei- der and Schneider (2010) found water temperatures up to 21.9 °C, at the end of breeding season (June). Bogaerts et al. (2010) report of a temperature drop of 2.5 °C from 8.3 °C to 5.8 °C within one week at the start of the breeding season in April, which did not seem to change the breed- ing activity. From a spring, the water temperature was only 8.9 °C, but after flowing through a completely de- forested and heavily grazed valley, the temperature rose about 2 °C per 100 meters up to 19 °C. Nevertheless, this wide temperature range is tolerated by N. strauchii, with Table 1. Mean lengths and weights of adult N. strauchii (Adapted from Pasmans et al. 2006). Data were collected in the breeding season. There is no significant difference between the subspecies or males and females between the subspecies (/-test). Subspecies Sex and number Mean total length (mm) Min - max total length (mm) Snout vent length (mm) Tail length (mm) Mean weight (g) barani Males (n = 11) 143 132-153 72 71 11.2 barani Females ( n = 25) 154 134-174 76 78 14.0 strauchii Males (n = 10) 143 131-176 73 68 10.3 strauchii Females (n= 17) 150 129-181 75 75 12.7 amphibian-reptile-conservation.org 011 September 2012 | Volume 6 | Number 4 | e53 Bogaerts et al. Figure 4. Habitat of N. s. strauchii near Bitlis. Photo by Serge Bogaerts. the wanner areas probably only increasing the develop- ment rate of larvae and shortening or shifting the aquatic phase in the adults. The streams in which the newts were found by Pasmans et al. (2006) were all slightly alkaline (pH 7-9) and soft to moderately hard, but these values can be strongly influenced by heavy rains or periods of prolonged drought. Nenrergus strauchii has been found overwintering on land, not far from streams (Schmidler and Sclnnidtler 1970). Adults, subadults, and juveniles have also occa- sionally been found under stones on land in April (Pas- mans et al. 2006). As streams probably partly diy, it seems likely that N. strauchii spends most of the year on land under stones or underground, protected from high temperatures and arid summer conditions. Breeding ani- mals in streams and pools are found during a relatively short period in spring from April to June (Steinfartz and Schultschik 1997; Bogaerts et al. 2010; Schneider and Schneider 2010). Protection Neurergus strauchii is a strictly protected species (Ap- pendix II) by the Convention on the Conservation of Eu- ropean Wildlife and Natural Habitats (also known as the Bern Convention), which was ratified by Turkey in 1984. In Resolution No. 6 (1998) of the Standing Commit- tee, N. strauchii is listed as a species requiring specific habitat conservation measures. The status of N. strauchii in Turkey is not clear, although the IUCN lists them as Vulnerable Blab (iii) (Papenfuss et al. 2009). Their cur- rently known distribution is much larger than previously thought, but the fact that they live in a habitat that is sensitive to human influences, and particularly climate change, makes them vulnerable. Habitat changes and destruction including overgrazing, pollution of breeding waters, cutting of trees, appear to currently be the ma- jor threats to the species (Bogaerts et al. 2006; Schneider and Schneider 2010). Materials and methods Origin of N. s. strauchii The origin of the N. s. strauchii being kept by the authors has an interesting history, as it involves extensive co-op- eration between privates and the N. s. strauchii originat- ed from a very small gene pool. Henry Janssen was one of the first people who succeeded in breeding F3 and F4 animals from captive breeding groups started by Fleck (FI) and Haker (F2) originating from Bitlis, near Fake Van, Turkey. These were distributed among other private breeders, including all authors on this article. Gunter Schultschik had several successful breedings (2000, 2001), and in 2003 Gunter bred a large group of offspring many of which were distributed within Europe, with a group being exported to the United States of America. All N. s. strauchii we have kept are direct descendants of the first breedings by Fleck. So we conclude that all ani- mals of this subspecies kept by the authors originate from the same very small gene pool and we have bred to at least the F5 generation. Most N. s. barani that are in cap- tivity originated from small private importations in 1997 and 1998, and two larger importations in 2002 and 2003. amphibian-reptile-conservation.org 012 September 201 2 | Volume 6 | Number 4 | e53 Conservation, husbandry, and captive breeding of the Anatolia newt, Neurergus strauchii Housing for adults in captivity Adults may be housed under a wide variety of condi- tions. The first main variation in housing is whether they are kept in an aquatic habitat all year or kept terrestrially for part of the year. Although in nature they will probably spend the majority of the year on land away from the breeding waters, some are kept aquatic for most of the year, or permanently. Different types of tanks are used for housing and rela- tively small: 30 x 40 cm to 50 x 120 cm. Individual carers use different furnishings for their terrariums. Terrestrial enclosures are often typical naturalistic terrariums with, for instance, a well-drained forest soil or loam and pieces of bark, moss, and plants to create shelter. Gunter Schul- tschik keeps his animals in a more sterile enclosure, on a five cm layer of synthetic foam, with shelters made out of pieces of bark. In this case, each tank is connected to a water system that drips cold water into the tank slowly, and seeps through the foam, running out again through a drain. This system works well in a warm and dry envi- ronment, but not in a relatively cold moist cellar or base- ment. When kept terrestrially, in a naturalistic enclosure, a water bowl is always present, and a gradation of humid- ity is offered so animals can choose from slightly humid to dry parts of the habitat. When kept all year round in an aquarium or aqua- terrarium, all carers provide the newts with an oppor- tunity to climb to a dry area, which usually consists of stone plates that are above the water level (Fig. 5). These stones are often covered with cork bark or some- times moss for hiding opportunities. The newts usually don’t remain in the dry region for long periods, only for a few hours or occasionally for a few days, except when temperatures rise above 20-22 °C, then they escape the water. Henry Janssen notes that in colder periods, with temperatures below 10 °C, the newts spend most of their time on land. Temperatures can drop in winter to close to zero and in summer can rise up to 30 °C. Animals that are kept aquatic during summer will typically stay in the water until the temperature of the water exceeds 20-22 °C. Incidental high temperatures of up to 30 °C do not directly harm the newts, as long as the newts are healthy and can stay on land. For lighting, natural light or fluorescent lamps are used. Temperatures in the tanks usually follow the sea- son in order to mimic the animals’ natural enviro nm ent (Table 2). Neurergus strauchii are very good at escape and will soon notice any chance to escape and take it. Therefore, it is necessary to cover the aquarium or ter- rarium with a secure, well- ventilated cover. Temperature cycling A cold period occurs in nature from autumn to spring, in the snow covered mountain areas where these newts live. In captivity, this cold period is simulated using different methods as part of the natural reproductive cycle. Half of the current authors hibernate their animals in a refrig- erator, approximately from mid-December to the end of February, at temperatures from 2 to 5 °C or at a constant 4.5 °C. Newts are kept in small boxes with wet paper towel(s) and bark with the sexes separated. The other half of the current authors keep newts under a regional temperature cycle at temperatures varying between 0-10 Figure 5. Aquarium constructed for N. s. strauchii. Photo by Jennifer Macke. amphibian-reptile-conservation.org 013 September 2012 | Volume 6 | Number 4 | e53 Bogaerts et al. Table 2. Mean temperature ranges in the adult environment through the seasons and aquatic (a), aqua-terrestrial (a-t) or terrestrial (t) set up of the tank. Keeper Spring Summer Autumn Winter Steinfartz (1995) 14 °C (a) Up to 23 °C (a) 10-14 °C (t) 10-14 °C (t) Henry Janssen 10-17 °C (a) up to 25 °C (a-t) 10-17 °C (a-t) 7-13 °C (a-t) Jennifer Macke 16-17 °C (a) 18-22 °C (a) 16-17 °C (a) 2-12 °C (a) Gunter Schultschik 16-17 °C (a) up to 30 °C (t) 16 (t) 4.5 °C (t) Kristina Ernst 12-18 °C (a) 18-27 °C (t) 8-18 °C (t) and more humidity 2-5 °C (t and a) Frangois Maillet 12-14 °C (a) 17-20 °C (a) 12-16 °C (a) 6-10 °C (t) Christoph Bork 12-16 °C (a) 17-21 °C (a), max. 25 16-19 °C (a) < 10 °C (t, for 2 months) Serge Bogaerts 12-16 °C (a) Up to 30 °C (a-t) 15-20 °C (t) 5-10 °C in a refrigerator (t) Patrick Wisniewski 10-15 °C (a) 15-25 °C (a) 15-20 °C (t) 10-15 °C (t) °C in garages, basements, or garden sheds for one to three months. Fleck (1982), Haker (1986), and Steinfartz (1995) all kept these newts in an unheated room where temperatures could drop as well. This is either done in terrestrial or aquatic conditions, and both sexes are usu- ally kept together. Newts can be transferred into another tank or stay in the same tank. Newts are mostly not fed during the cold period; only Jennifer Macke feeds them twice per week throughout the cold period and finds that they eat well, and are active even when their temperature is as low as 2 °C. Diet and nutrition Adult newts eat many types of living and non-living food. On land we offer them a wide variety of insects, including young crickets ( Acheata domesticus or Gryllus sp.), mealworms ( Tenebrio molitor), fungus beetle lar- vae ( Alphitobius laevigatas), and larvae of wax moths; both the lesser ( Achroia grisella ) and greater ( Galleria mellonella). We also feed earthworms ( Lumbricus sp.), maggots, firebrats/silverfish ( Thermobia sp.), and slugs. In water they are fed earthworms, black worms (Lum- briculus variegatus), Tubifex sp., bloodworms ( Chironi - mas sp.), Daphnia sp., Gammarus sp., Hyalella azteca, white worms (Enchytraeus albidus), woodlice ( Asellus sp.), etc. Amphibian eggs and larvae ( Rana sp.) are eat- en. Henry Janssen also saw them eat small fish (Gup- pies, Poecilia reticulata) at night when the guppies were sleeping. Non-living prey is accepted. Fleck (1982) fed them slices of liver, and Christoph Bork fed them, with tweezers, octopus that was cut into small worm-like strips. Kristina Ernst reports that keeping females on land makes it easier to give high calorie food like wax worms, which seems to yield more eggs the next breeding peri- od. Henry Janssen has noted that, with equal amounts of food offered, juveniles grow faster and adults gain more volume at lower temperatures (10-17 °C) than at higher temperatures (18-25 °C). Neurergus strauchii is not as voracious a feeder as, for instance, newts of the genus Triturus. Neurergus strauchii may be rather slow to catch prey. Neurergus s. barani seem to be more greedy for food and eat everything in greater portions, compared to N. s. strauchii in our experience; it is one of the few sig- nificant differences between keeping N. s. strauchii and N. s. barani. We find that feeding plenty of (high calorie) food during the breeding period is essential for females to produce many eggs. Food items offered on land are typically dusted with a calcium vitamin powder. We have used, for example, Korvimin ZVT, Amivit A, Nutrobal Vitamin/Mineral powder, and ZooMed Calcium. Gut loading crickets with calcium rich plants, like dandelion, or nettles will en- rich their food quality. Feeding crickets at temperatures below 10 °C is difficult as most crickets die. Individual newts can have very different preferences for food items. Results Breeding For breeding purposes, the newts are placed into an aquarium. The tanks are furnished in various ways. Most of us use a layer of gravel on the tank floor, and various types of stones are placed on top of each other to provide places for hiding and egg deposition. Jennifer Macke uses turned over non-glazed ceramic flower pots with a cut out entrance, used by the females to deposit their eggs, which can easily be taken out with the eggs and replaced. Some of us have used no substrate or just a few flat stone plates, covering only part of the tank bottom. Tables 3 and 4 report the periods, temperatures, and other characteristics of the various breeding tanks. Development of enlarged cloacas and the whitish-blue colorations on tails of males can already be observed in autumn. The smallest male in captivity bred measured 11.5 cm total length (TL) and 6.2 cm snout-vent length (SVL); the smallest female measured 12.8 cm TL and 6.5 cm SVL. Thus, animals start breeding at total lengths of around 12 cm TL. Breeding occurs within a water temperature range of 9-17 °C (mean 10-14 °C) and this seems to be independent of the time of the year (Table amphibian-reptile-conservation.org 014 September 2012 | Volume 6 | Number 4 | e53 Conservation, husbandry, and captive breeding of the Anatolia newt, Neurergus strauchii Table 3. Aquarium conditions when breeding started. Included are only those years in which fertile eggs were deposited. Keeper Subspecies Year Starting Temperature start breeding (°C) Water level (cm) Water circulation and/or air pump Fleck (1982) strauchii 1981 March 12 10 yes Steinfartz (1995) barani 1993-1994 Feb-March 14 25 yes Jennifer Macke strauchii 2005-2009 2011-2012 Dec-Jan 9-12 20 yes Gunter Schultschik strauchii & barani 2003-2004 Jan-Feb 16-17 28 yes 2004 May May April 14 Kristina Ernst strauchii & barani 2005 2011 14 14 11-12 yes 2012 Feb 10-12 Francois Maillet barani 2005 March- April 12-14 12/15 yes and air pump Christoph Bork strauchii & barani 2001 2003 2005 Feb 13-15 24-28 yes Patrick Wisniewski strauchii 1996 1997 Feb-March 10-12 15 strong air pump only Serge Bogaerts strauchii 2006 Feb 12-14 8 yes 3). Newts were bred in winter, early spring or even to the end of spring. The water level does not seem to be important. As these newts are stream dwellers, most of us have simulated this by using water circulation, some- times with the addition of an air pump. Breeding starts with male activity, typically at water temperatures of 10 °C. Males and females can be put in the water at the same time, but some of us prefer to intro- duce females to the water a few days or weeks later. Af- ter entering the water, males have been observed to start performing courtship the same evening. Within the court- ship period, it is best to try and keep water temperatures below 14 °C. At 14 °C females start oviposition (Table 4 and Fig. 6). Figure 6 shows oviposition in three of the most suc- cessful breeding years, in relation to the water tempera- ture. Oviposition may take place during both day and night and may continue until water temperatures reach about 20 °C. Eggs are laid very irregularly in time and number, and oviposition may depend on the condition of the female, particularly her nutritional condition through diet. Occasional egg laying (one per day or less) can continue for up to two months after the main period of oviposition. Henry Janssen measured the water temperatures at which oviposition took place for 1,225 eggs from dif- ferent breedings over the years 1991-1997. He also noted which of these eggs hatched. Of all eggs, only six (0.48%) were laid at water temperatures below 14 °C. Most eggs (77.3%) were deposited at temperatures of 15-19 °C. Above 20 °C, production of eggs rapidly de- creases. Figure 1 shows a dip at 16-17 °C, but we think this is an artifact of the combination of data from differ- ent years. Another finding of Henry Janssen is that of all eggs that were deposited, the ones laid between 14-16 °C had the best hatching rate (62.4% between 14-15 °C and Figure 6. Oviposition (n = 760 eggs) in three successful breed- ing years in relation to water temperature. Data by Henry Jans- sen. 28.5% between 15-16 °C). There are several possible in- terpretations for these data. First, it may be related to the fecundity of the females; the first eggs laid are often of a higher quality than later eggs. Second, it could be related to the fertility of the males, which seem to be more active at lower temperatures. Jennifer Macke has also noted that egg fertility consistently decreases over time during the egg laying period (data not shown). Henry Janssen noted from the 1995 breeding season that when he separated females from males, after he discovered that males were eating some of the first eggs, most eggs laid afterwards were not fertilized. This seems to indicate that regular uptakes of spermatophores by the female, during the breeding period, are necessary for her to continue pro- ducing fertile eggs. Table 4 records the aquarium conditions when fe- males started oviposition — the number of eggs per fe- male, and the percent of hatched eggs. As can be seen, large variations were found in number of eggs per female amphibian-reptile-conservation.org 015 September 2012 | Volume 6 | Number 4 | e53 Bogaerts et al. Table 4. Conditions in the aquarium when females started oviposition, number of eggs per female (~ when more females are kept to- gether), time to metamorphosis, and percent hatched. - No data available. # Average over the whole oviposition period. *Ten of these are 14 months old but still have not completed metamorphosis; they show no differences in length compared with their siblings. Number of Time to meta- Keeper Subspecies Year Starting T°C eggs per morphosis Hatched female (months) Fleck (1982) strauchii 1981 April 17 ~ 75 4.5 - Haker (1986) strauchii 1985 June 16 - 3 - Steinfartz (1995) barani - - - 80-90 - - 2005 Feb 16 14 152 - -50% 2006 Feb 19 13 150 - -50% 2007 Feb 27 - 104 - - Jennifer Macke strauchii 2008 March 4 - 246 - - 2009 Feb 27 - 285 - - 2011 Feb 26 - 238 - mean 41% # 2012 March 8 - 195 - mean 78% # 2004 May 14 Gunter Schultsckik strauchii 2005 2011 May April 14 14 11-12 yes 2012 Feb 10-12 barani 2004 June 16 -200 5-8 - 50% barani 2005 May 20-21 - 100 4-7 - 25% strauchii 2005 May 17-19 - 150 4-7 98% Kristina Ernst barani 2006 May 16 -250 - - 75% strauchii 2006 May 17 -200 - 97% strauchii 2011 April 15 - 150 4-7* 88% strauchii 2012 March 15 - 100 - 80% barani 2012 April 15 - 100 - 90% Christoph Bork strauchii & barani 2001 2003 2005 March 14-16 - - 4 No counts, but never 100% strauchii 1992 Mar-April 14-17 129 4-8 45% Henry Janssen strauchii 1995 April-May 16-19 4-8 strauchii 1996 April 16-17 -85 4-8 25% Patrick Wisniewski strauchii 1996 March 10-15 47 5-6 45% strauchii 1997 February 10-15 17 5-6 50% Serge Bogaerts strauchii 2006 March 14-16 -40 5-10 70% and hatching rates. However, the temperature conditions in which oviposition occurred were roughly the same for all of us, for both subspecies. Eggs Eggs are mostly attached to the underside of stones (Fig. 7), but they can be laid almost anywhere, including on the filter, aquarium walls, and plants (Fig. 8), or specially prepared flower pots. Eggs may be found loose on the bottom of the tank, but this mainly occurs when there is too little space on the favorable places and, or eggs are not well attached. During oviposition the female lies on her back, often sandwiched between two layers of flat rock, depositing eggs on the underside of the upper rock. It is important that the habitat has enough space between the stone plates for the females to move around. Henry Janssen noted that out of a total of 560 eggs, 237 were deposited on the glass, 199 on stones, 83 loose on the substrate, 37 on plants, three on the filter system, and one was stuck to the hind leg of a female. No negative effects from exposure of developing eggs to indirect sunlight or artificial light could be observed when compared to eggs that developed under darker conditions. In general, eggs were removed from the breeding tank, as the adults sometimes eat the eggs. Eggs were typically removed every few days. Some of us moved the eggs together with the stones they were attached to, oth- ers cut the eggs gently loose from the rocks with a razor blade or fingernail. There was no difference observed in the development of eggs that were cut loose versus eggs that were left on the stones they were laid upon. Water parameters of the tank, where the eggs are put to hatch, do not seem to matter. Even an air stone is not really necessary for the development of the eggs. If the water is refreshed once a week this seems to be enough. It is, however, also possible to leave the eggs in the tank until they hatch, which some of us prefer. In all our breedings, no clutch of eggs was 100% fertile. Unfertilized eggs and eggs that have died off, shrink in size and start decaying, resulting in the clear layers around the zygote becoming cloudy, starting with amphibian-reptile-conservation.org 016 September 201 2 | Volume 6 | Number 4 | e53 Conservation, husbandry, and captive breeding of the Anatolia newt, Neurergus strauchii Figure 7. Female N. s. strauchii depositing eggs. Photo by Christoph Bork. the innermost layer and continuing outwards, followed by mould — growth on the outer surface (observations Henry Janssen; Fig. 9). It seems that a developing egg, attached to a moulding egg, can be infected with fungus too. Therefore, it is best to separate moulding eggs from developing eggs. Eggs can be eaten by the usual preda- tors like snails and flat worms ( Planaria sp.). Hatching success of eggs can vary enormously (Table 4). Jennifer Macke noted that in 2005 about 50% of eggs were fertile, and 10% began to develop but died as embryos. In 2006 about 80% were fertile and about 10% began to develop but died as embryos. Malformations seem to occur in all breedings. Kristina Ernst states that through feeding the females more often, more eggs are produced and in shorter periods. She observed up to about 15 eggs per day per female. In 2005 Jennifer Macke had eggs laid from February until the beginning of March. After that animals were transferred to another location and they continued to lay eggs (about one a day), but all eggs produced in April and May were infertile. In 2009 Jennifer counted a total of 570 eggs from two females during the entire egg lay- ing period (February- June). Henry Janssen measured the hatching success of all eggs deposited between 1991 and 1997, each year breeding occurred. Of the total of 1,413 eggs, 348 hatched (24, 62%). Gunther Schultschik noted the exact water parameters in his rearing tanks. Larvae were raised at a water tem- perature of 16-19 °C, with no measurable organic ions in the water (NH 3 , N0 2 , N0 3 ), maximum of oxygen, mini- mum of CO,. Water was treated by UV lamp. PH was 7.2 to 7.5. Francis Maillet maintains a pH of 7-8 and changed part of the water often to avoid nitrate develop- ment. Henry Janssen measured the length at hatching for 283 specimens (Figure 10). About 45% of the measured larvae were between 12 to 14 mm at hatching. Fig. 11 shows a hatching larvae. Henry Janssen measured the relationship between days of incubation and total length at hatching for 249 larvae. The shortest time to hatch was 15 days and the longest was 34 days. About 57.4% of all larvae hatch be- tween 26 and 3 1 days after deposition. Consistent with this, Jennifer Macke found that from the time the first eggs were laid until the first larva hatched, exactly 30 days elapsed when the eggs were maintained at 16-17 °C. The total length of the larvae becomes larger when hatching is delayed. Thus, the moment of hatching is not a fixed point in time. Moving the egg, for instance, can cause the larva to leave the egg shortly thereafter, where- as it would have stayed in place if the egg had been left undisturbed. Larval rearing All authors raised their larvae in more or less the same way, and all agreed it was not very difficult or problem- atic. For the first few days after hatching, the larvae live on their yolk. No food was added at this time, and some * $ % # f/i J J i f/M 5r Figure 8. Fresh laid N. s. strauchii egg. Photo by Henry Jans- sen. amphibian-reptile-conservation.org 017 September 2012 | Volume 6 | Number 4 | e53 Bogaerts et al. Figure 9. Moulding unfertilized eggs of N. s. strauchii on mm paper Photo by Serge Bogaerts. Figure 10. Total length of larvae at hatching in mm. Data by Henry Janssen, n = 283. authors noted that micro-organisms, particularly water mites ( Hydracarina sp.) and Cyclops sp. attacked new- ly-hatched larvae. After a few days the larvae begin to eat live food. Larvae are kept in tanks or tubs containing three to 20 cm of water, an air stone, and some pebbles and pots as hiding places. Aquatic plants are sometimes included. Larvae are fed first with Artemia (only the first one or two weeks), small live Daphnia sp., Tubifex/Lum- briculus (initially chopped, later whole), red mosquito larvae/bloodworms ( Chironimus sp.), and white worms ( Enchytraeus albidus ). Gunter Schultschik gave Artemia until the larvae were 20 mm. When feeding Daphnia, care must be taken to avoid feeding other less harmless aquatic fauna. Water temperatures can range from 10 to 20 °C. Even if the temperature of the water rises up to 30 °C accidentally, it is not a serious problem, although larvae stop eating and become less active. The larvae are not as aggressive toward each other as, for instance, Triturus larvae, but care must be taken to avoid overcrowding. Kristina Ernst noted cannibalis- tic behavior until the larvae were 1. 5-2.0 cm, at which point the behavior disappeared. Several of us have never observed cannibalism and even kept larvae of different sizes together without a problem. Most of us have kept the larvae in small groups (15-30 larvae) in, for instance, plastic containers of various sizes with aquatic vegeta- tion and shelters, like pieces of ceramic garden pots, as these salamanders hide during the day. Water is refreshed every week, or as often as required to avoid poor water quality. In some cases, malformed larvae hatch. These larvae spin around when trying to swim, or are swollen. These larvae often lag far behind their siblings in growth, and euthanasia is the best option. Larvae of a few centime- ters in size develop gold colored, shiny spots and dots that seem similar to the lateral line sense organs in fish used to detect movement and vibration in surrounding water (Fig. 12), which stay visible until metamorphosis (Fig. 13). After about three to four months the larvae de- Figure 11. Hatching larvae of A. s. strauchii. Photo by Serge Bogaerts. amphibian-reptile-conservation.org 018 September 2012 | Volume 6 | Number 4 | e53 Conservation, husbandry, and captive breeding of the Anatolia newt, Neurergus strauchii Figure 12. Larva of N. s. strauchii few weeks old, the lateral line sense system visable in stripes on lateral sides and tail and in spots behind the eye. Photo by Serge Bogaerts. Figure 13. Larva of N. s. strauchii of approximately four months old. Photo by Serge Bogaerts. velop yellow spots and later become darker and darker developing their juvenile black pattern (Figs. 14, 15). Another one to three months may elapse before the gills are completely gone. Mortality of larvae is very low. Lar- vae become lighter in color at night. Depending on the water temperature and the amount of food, larvae meta- morphosed in three to seven months, with a mean period of about five months (Table 4). The first shedding takes place at around the time of metamorphosis, sometimes just before emergence from the water. They leave the water mostly at night and search for a hiding place, and if not provided, they try to hide again in the water. The first few weeks after meta- morphosis, the juveniles can be kept in an aqua-terrarium with different hiding places from wet to dry, from which they can choose. Metamorphosis in this newt seems to be very gradual, such that juveniles continue to shift from water to land during a period of several weeks. After metamorphosis the juveniles resemble their par- ents, although they have significantly fewer yellow spots, and spots are confined to two rows along their backs. The bellies are not completely black and show light-colored parts. The orange-red stripe on the belly is rose-orange and not as brilliant as in the adults. We are positive color intensity in captive-raised adults depends on the amount of carotenoid-rich food animals eat, like in the Japanese fire belly newt, Cynops pyrrhogaster (see Matsui et al. 2003). Henry Janssen measured the total length of 108 speci- mens at the moment of metamorphosis (Figure 16). The amphibian-reptile-conservation.org 019 September 2012 | Volume 6 | Number 4 | e53 Bogaerts et al. Figure 14. Larva of N. s. strauchii change its coloration to juvenile pattern. Photo by Serge Bogaerts. Figure 15. Larva of N. s. strauchii just before metamorphosis. Photo by Serge Bogaerts. data include only larvae that metamorphosed within the year eggs were laid. Metamorphosed N. s. strauchii weigh about 0.60 g (n = 11, with mean total length of 55 mm; data Serge Bogaerts) which corresponds to Schult- schik (data not shown) who gives 0.67 g for metamor- phosed N. s. strauchii. Henry Janssen measured the rate of metamorphosis of all eggs deposited between 1991 and 1997 in which breeding occurred each year. Of the total of 1,413 eggs, only 138 specimens reached metamorphosis (9.8%) (see Table 5). Metamorphosis was considered as the moment the gills disappear, the black and yellow coloration are vis- ible, and juvenile newt(s) come onto land for the first time. However, this is not a fixed moment. They can stay in a semi-aquatic stage for a while, with very short gills and full black and yellow coloration. The data of Hen- ry Janssen show that there is a wide range of lengths at which metamorphosis can take place (Fig. 16). All of the measurements taken by other breeders have fallen within these ranges (Table 6). “Overwintering” larvae In both N. s. strauchii and N. s. barani, overwintering larvae are observed. Larvae that hatch later in the season, or stay behind in development, will remain larvae dur- ing the winter and metamorphose the next year. Fleck (1982) and Haker (1986) describe N. s. strauchii still found in larval fonn in January. Pasmans et al. (2006) describe this phenomenon for N. s. barani. During a field visit in May 2006, special attention was paid to this phe- nomenon at the type locality of N. s. barani, and many larvae that hatched in 2005 could be observed (S. Bo- gaerts, pers. obs.). The larvae keep their gills and fins, but develop characteristics of the juvenile coloration: black background color and yellow spots. Overwintering larvae seem to grow a bit larger than their siblings that amphibian-reptile-conservation.org 020 September 2012 | Volume 6 | Number 4 | e53 Conservation, husbandry, and captive breeding of the Anatolia newt, Neurergus strauchii Table 5. Survival rate from egg laying until metamorphosis over seven years of breeding. Data by Henry Janssen. Year Number of eggs Hatched eggs Metamorphosed Success rate per year (%) 1991 40 0 0 0 1992 514 229 48 9.3 1993 85 30 24 28.2 1994 39 2 2 5.1 1995 476 43 30 6.3 1996 171 43 34 19.8 1997 88 1 0 0 Table 6. Lengths of larvae at metamorphosis. Keeper Subspecies Length (mm) Remarks Schmidtler and Schmidtler 1970 strauchii 54-61 Fleck 1982 strauchii 54-55 Steinfartz 1995 barani 56 Henry Janssen strauchii 47-75 Gunter Schultschik strauchii 45-50 Kristina Ernst barani & strauchii 40-60 Jennifer Macke strauchii 60-65 Still with gills Francois Maillet barani 55-60 completed metamorphosis the previous year (up to 75, 25 Jennifer Macke tested the difference between terres- mm; data Henry Janssen). Raising juveniles Fleck (1982) writes that raising juveniles is not problem- atic, as they easily switch between aquatic and terrestrial living, and can be kept and raised in an aqua-terrarium. Most of us raise the juveniles terrestrially. This method of rearing is most like their natural conditions, where ju- veniles live terrestrially until reaching reproductive age. A small terrarium (50 x 20 x 15 cm) is often used, with a leaf litter soil (typically from beech or oak forest), or a mixture of substrates (soil, coconut fibre, etc.) and some pieces of bark, which the newts use as shelter. A more sterile option with moist paper towel(s) and some pieces of bark also works well, but needs cleaning at least once a week. The juveniles are fed at a minimum of once a week, or usually more (further details above under the “Diet and nutrition” section on page 14). Tanks should provide a range of dry and moist places (Fig. 17). Frank Pasmans raised juveniles on wet Kleenex kitchen towel paper, with pieces of ceramic roof tiles piled up, creating gradients from moist to dry. Foods are prepared similar as for the adults and are typically small crickets, small wax wonns, slugs, fruit flies {Drosophila sp.), woodlice {Asellus sp.), firebrats/ silverfishes ( Thermobia sp.), etc. Further, bloodworm (Chironimus sp.), Tubifex sp., or chopped earthworms and blackworms can be fed from a small bowl or on a wet paper towel. They can be kept in the same temperature ranges as adults. Our captive bred animals have reached at least the age of 12 years. trial and aquatic raising of juveniles. In March 2004, four of the juveniles obtained in October 2003 were adapted to water at a size of about 7-8 cm total length. This was accomplished by placing them, one at a time, into an 1 8-liter (five gallon) tank containing two cm (one inch) of water, a thick layer of aquatic plants, and an ample supply of live blackwonns {Lumbriculus variegatus), and chopped earthworms. Each animal adapted to hid- ing beneath the plants within one day. Once adapted to water, they were moved to a larger tank (60 x 30 x 30 cm) containing 25 cm of water, large river rocks, clay pots, and a mini canister filter providing a bit of current. Local tap water comes from ground water that is alkaline and moderately hard (GH 70 ppm, KH 90 ppm, pH 8). Both the aquatic and terrestrial animals appeared healthy and grew well. Feeding regimens were, of necessity, dif- Length at metamorphosis 14 12 10 a> 8 .o E 3 C 6 4 2 1111 . n _.UL_D 0 20 °C). A six week quarantine period is recommended when having first obtained animal(s). During this period, the newly acquired animal should be assessed by a qualified veterinarian for the presence of infectious and non-infec- tious diseases. We strongly recommend every newly ac- quired animal to be tested for the presence of ranaviruses and Batrachochvtrium dendrobatidis . The presence of both agents can be assessed by detection of their respec- tive DNA in skin swabs (less sensitive for the detection amphibian-reptile-conservation.org 022 September 2012 | Volume 6 | Number 4 | e53 Conservation, husbandry, and captive breeding of the Anatolia newt, Neurergus strauchii Figure 18. Adult of A. s. strauchii of “Gold-dust” form. Photo by Henry Janssen. Figure 19. Metabolic bone disease in an adult female N. s. strauchii. Note the malformation of the lower jaw. Photo by Frank Pasmans. of ranaviruses) or tail clips. Trade derived animals have indeed been identified as important carriers of both infec- tious agents and may spread diseases to native amphibian populations. Both diseases have been listed by the Office International des Epizooties or World Organisation for Animal Health (OIE) as notifiable diseases since 2008. The following disorders have been diagnosed in Neurer- gus (in part by F. Pasmans, pers. observ.): 1) Metabolic bone disease (MBD, Fig. 19). MBD comprises a number of metabolic disorders affecting skeletal calcification. In urodelans, most cases of MBD can probably be attributed to relative lack of calcium and/or vitamin D in the feed, and would thus be more appropriately named, nutritional secondary hyperpara- thyroidism. Clinical signs are most obvious in young, terrestrial specimens, and include backbone and head malformations (e.g., shortening of the lower jaw), and abnormal movements. MBD can be prevented by supply- ing feed items (e.g., crickets) with extra calcium through the insect diet (“gut loading”) and topically applying cal- cium containing powder on the feed insects. However, this is only applicable for juveniles raised on land and for terrestrial adults. Feeding calcium supplementation for aquatic newts is much more difficult to achieve and may in part be met by providing calcium supplemented pellet feed (e.g., turtle pellets, if accepted by the newt). 2) Ranavirosis. Recently, ranavirosis has been de- scribed in N. crocatus, imported from Iraq (Stohr et al., in prep.). Clinical signs of this viral disease include red- dening of the skin (erythema), skin ulceration, edema, anorexia, and death. The course of a Ranavirus infection may vary from subclinical (without clinical signs) to mass mortality. This virus is one of two known infectious threats to amphibian biodiversity worldwide. Prevention consists of quarantine measures of newly acquired ani- mals and preferably testing of a tail clip or skin swab for the presence of the viral DNA. It is of utmost importance to prevent any contact of Ranavirus -infected newts or their enviromnent (e.g., aquarium water) with the envi- ronment, to prevent spread of the virus to native amphib- ian populations. Ranavirosis cannot be treated. 3) Chytridiomycosis. This fungal disease is caused by Batrachochytrium dendrobatidis and is considered the most important infectious driver of worldwide amphib- ian declines. For this reason, it is of utmost importance that (as for ranavirosis), captive populations of Neurer- gus are negative for the fungus. It is at present not clear whether this fungus causes clinical problems in newts of the genus Neurergus. In other amphibians, the course of a B. dendrobatidis infection may vary from asymptomatic to apathy, skin disorders, and death. Recently, B. dendro- batidis infection was demonstrated in N. kaiseri (Spitzen van der Sluijs et al. 2011) but no clinical signs of disease were noticed. As a preventative measure, all newly ac- quired Neurergus should be tested for the presence of the fungus using a skin swab. If positive, infected animals and their captive environment should be treated appro- priately. Neurergus kaiseri was treated successfully us- ing voriconazole (F. Pasmans, pers. observ.; Martel et al. 2011). As for ranaviruses, all contact of B. dendrobatidis infected animals and their captive environment with the outside environment should be strictly prevented. amphibian-reptile-conservation.org 023 September 2012 | Volume 6 | Number 4 | e53 Bogaerts et al. Figure 20. Ascites (“bloating”) in an adult female N. s. strauchii with a severe enteritis, associated with high numbers of flagellates. Photo by Frank Pas mans. 4) Chlamydiosis. For more than a decade, enigmatic mortality in captive N. crocatus and N. strauchii newts was observed by several breeders. This mortality even impaired the establishment of successful breeding pro- grams in for example N. crocatus. In the nineties of the past century, entire captive breeding groups of this newt were lost. Keepers reported non healing wounds on the tail. Recently, the cause for this mortality was suggested to be a bacterium: Candidatus Amphibiichlamydia sal- amandrae (Martel et al. 2012). The disease presents as anorexia, lethargy, edema, markedly abnormal gait, and death. Secondary bacterial or mycotic infections (e.g., with Aeromonas sp. or Mucor sp.) appear to be common. Urodelans can be very probably latent carriers of Chla- mydia bacteria, with possible reactivation of the infection during stress periods. Indeed, Chlamydia infections are probably widely spread in urodelan collections and clini- cal signs are possibly provoked by suboptimal condi- tions, for example, elevated temperatures during summer months. Until now, clinically infected animals invariably die but therapy may consist of the use of, for example, tetracyclines to cure the infection. Preventative measures consist solely of quarantine measures and optimal hus- bandry (including temperatures <20 °C). 5) Intestinal parasitosis (Fig. 20). As in all amphib- ians, intestinal parasitosis may occur in Neurergus newts and appears to be mostly provoked by suboptimal hus- bandry. Several cases of severe enteritis, coinciding with very high numbers of flagellate protozoa were diagnosed in N. strauchii and N. crocatus. Clinical signs were an- orexia, loss of condition to cachexia, and in some cases ascites (bloating). Treatment using metronidazole and optimizing husbandry was successful in cases with an early diagnosis. Conclusions In addition to their beauty, N. strauchii are interesting newts in captivity. Although our data are still scattered and incomplete, the results of this project presents good indications for long-tenn captive maintenance and gives direction for further studies especially, when our experi- ences differ, or have revealed new topics to study. Our main goal in keeping this newt has been its suc- cessful breeding. We vary in our opinions about the importance of a terrestrial period as part of the yearly breeding cycle. Although, it is in their natural cycle to have a terrestrial period both Fleck (1982) and Steinfartz (1995) write that a terrestrial phase is necessary to initi- ate breeding. Fleck kept his animals from 1975 to 1979 in an aquatic enclosure, but males and females were not synchronized in their breeding behavior. He gave three animals a terrestrial phase in 1980 and then both sexes got into breeding condition at the same time. However, Jennifer Macke has bred the same group of animals for eight years without any terrestrial periods, while most of us experienced that animals were less willing to breed when they were not given a terrestrial period. Thus, if keeping adults aquatic or not, the whole year round is not of major importance in breeding these ani- mals, we show that a dramatic change in temperature, often combined with an abrupt shift from land to water is probably more important. Steinfartz (1995) reported that in his opinion it is not a low temperature in win- ter that is necessary for successful breeding, but a strong temperature difference between summer and winter pe- riod. However, our findings indicate a strong correlation between specific temperatures and the specific phases of reproduction. A period with temperatures below 10 °C amphibian-reptile-conservation.org 024 September 2012 | Volume 6 | Number 4 | e53 Conservation, husbandry, and captive breeding of the Anatolia newt, Neurergus strauchii Figure 21. Proposed yearly temperature-curve for the captive breeding of Neurergus strauchii. prior to the breeding season proves essential to realize the temperature curve that stimulates breeding (Fig 21). Hence, a terrestrial period may help to synchronize the sexes and breeding behavior. Adults can be kept all year round aquatic and will still breed, but need to at least undergo a change in temperatures. Keeping animals in a terrestrial phase makes it easier to change temperatures by, for instance, placing them in a refrigerator or out- doors. Another advantage of a terrestrial period is that one can provide more differentiation in food items (e.g., crickets, wax worms, and other insects) which increased diet quality. It seems that changing the newts between enclosures is not a drawback. When animals are kept the whole year in the same tank, with just gradual changes of temper- ature, they will not breed, but if they have a dramatic change of environment (or change of temperature), they are likely to initiate breeding behavior. We all agree that giving the newts a cold winter period is the best way to have a successful breeding. Although Sparreboom et al. (2000) noted interfer- ing males when a couple is mating, it does not seem to decrease the success rate of breeding, but if undisturbed breeding is the goal, one should keep one male together with one or more females during the breeding period. For successful initiation of oviposition, it seems that a shift in temperature from 12-14 °C (courtship behavior and development of eggs) to 16-18 °C is important. A temperature stable environment where the tank is placed (basement) or refreshing the water can help provide this. The period of egg laying seems to depend on the gradual rising of temperature. If temperature rises quickly this can reduce, or even eliminate, the period of egg laying. Henry Janssen noted such negative influences on his breeding results, caused by rapid and unwanted tem- perature changes, typical for the sea climate where he lives. It seems therefore important for the breeder to have some control over water temperature. In nature, depend- ing on the weather, mating season starts approximately at the end of April to beginning of May for both sub- species (Bogaerts et al. 2006) and continues into June (Schmidtler 1994). In captivity, temperature is of more influence than the time of the year and newts are prepar- ing to breed when temperatures rise above 10 °C. Although eggs are mostly “spawned” on the under- side of stones, they can be laid anywhere, as described. Bogaerts et al. (2006) found some eggs of A. v. barani on tree branches and at the bottom of water bodies, but con- cluded it was an artifact due to the lack of other suitable sites. This might also be true for captive populations. Eggs laid or kept in lighter enviromnents develop in the same manner, and we have not found any differences in development. Development of eggs does not seem to be influenced by taking them out of the breeding tank or cutting them loose from stones. The eggs are not deli- cate. One can choose the way that suits the breeder best. Taking the eggs out of the tank gives the breeder more control. Our data show total number of eggs per female can be much higher, up to 285, than previously reported (Schmidtler and Schmidtler 1970; Fleck 1982; Stein- fartz 1995). It seems likely the amount of food given to females adds to production of more eggs. Even young females can lay many eggs, thus size of the female does not seem to be a major factor determining the amount of eggs laid, but the amount of food given probably is amphibian-reptile-conservation.org 025 September 2012 | Volume 6 | Number 4 | e53 Bogaerts et al. a contributing factor as well; however, we do not have enough data to support this claim. There doesn’t seem to be a difference between the two subspecies in the number of eggs. Steinfartz (1995) stated that in his opinion the lar- vae of N. s. barani are more pond-type, in comparison to stream-type larvae of N. s. strauchii, based on minor differences. In the experience of the authors that have kept both subspecies, it is impossible to tell larvae apart. However, perhaps N. s. barani grows a little larger and more robust before metamorphosis. But we do not have data to support this. Rearing of the larvae and juveniles was relatively unproblematic in all cases, as long as water quality is checked regularly. Raising juveniles can be done in a ter- restrial environment or in an aqua-terrarium. In nature, juveniles certainly grow up terrestrial, as aquatic juve- niles have never been observed in the field. Raising them in an aqua-terrarium gives the opportunity to feed them more varied types of food. The newts are able to switch easily from land to water and vice versa, and they adapt quickly to water without skin problems. Jennifer Macke showed a slightly faster growth of aquatically-raised animals. But as the number of animals is very small and because the feeding regime and types of food differed, it is not possible to draw straightforward conclusions. It seems likely that growth depends more on quality and availability of food and temperature, than on the type of housing. In the past, whole groups of captive bred N. strauchii have collapsed; also among the authors. Sev- eral possible and proven candidates have been described above. We strongly advise to avoid stress (e.g, high tem- peratures >20 °C) and provide optimal husbandry and feeding. We hope our successful long-term keeping, breeding, and raising of N. strauchii is an example and model that may be used for private contributions to conservation breeding programs, for endangered Neurergus species and other semi-aquatic salamanders. Future studies on captive specimens will provide more data on the cap- tive breeding of this, and other newts. According to the Studbook (Molch-Register) by Kristina Ernst it seems this species is still available in good numbers, but con- sistent breeding every year is still rare, even among the authors. Hopefully, this paper can contribute to greater captive breeding efforts and to a better understanding of the ecology of this fascinating newt but also can be seen as a valuable example of privates’ contribution to con- servation breeding programs for endangered Neurergus species and other semi-aquatic salamanders. Acknowledgments. — Henk Wallays is thanked for his support and encouragement in the early part of the proj- ect. We thank the AG Urodela (www.ag-urodela.de) and the Salamander Society (www.salamanders.nl) for their support. Literature cited Bogaerts S, Pasmans F, Woeltyes T. 2006. Ecology and conservation aspects of Neurergus strauchii (Am- phibia: Salamandridae). In: Herpetologia Bonnen- sis II. Proceedings of 13th Congress of the Societas Europaea Herpetologica. Editors, Vences M, Kohler J, Ziegler T, Bohme W. Societas Europaea Herpeto- logica. 15-18. Bogaerts S. 2007. Internationales Projekt: Haltung und Zucht des Turkischen Bergbachmolches, Neurergus strauchii. Amphibia 6( 1 ): 1 6- 1 7. Bogaerts S, Pasmans F, Sparreboom M, Biricik M. 2010. Observations on a breeding population of Neurergus strauchii barani Oz, 1994 (Caudata: Salamandridae). Salamandra 46(l):55-58. Fleck J. 1982. Erst-Nachzucht des Turkischen Berg- bachmolches, Neurergus strauchii. Salamandra 18(3- 4): 138-149. Haker K. 1986. Nachzuclit in zweiter Generation des turkischen Bergbachmolches, Neurergus strauchii (Steindachner, 1887). Salamandra 22(4):286-287. Macke J. 2006. Neurergus strauchii. [Online]. Available: http://www.caudata.org/cc/species/Neurergus/N_ strauchii. shtml [Accessed: 19 June 2012]. Martel A, Adriaensen C, Bogaerts S, Ducatelle R, Hae- sebrouck F, Pasmans F. 2012. Novel Chlamydiaceae disease in captive salamanders. Emerging Infectious Diseases 18(6): 1020-1022. Martel A, Van Rooij P, Vercauteren G, Baert K, Van Waeyenberghe L, Debacker P, Gamer TW, Woeltjes T, Ducatelle R, Haesebrouck F, Pasmans F. 201 1. Devel- oping a safe antifungal treatment protocol to eliminate Batrachochytrium dendrobatidis from amphibians. Medical Mycology 49(2): 143-149. Matsui K, Mochida K, Nakamura M. 2003. Food habit of the juvenile of the Japanese newt Cynops pyrrhogas- ter. Zoological Science 20(7):855-859. Oz M. 1994. A new form of Neurergus strauchii (Uro- dela, Salamandridae) from Turkey. Turkish Journal of Zoology’ 18:115-117. Ozdemir N, Uzurn N, Avci A, Olgun K. 2009. Phylog- eny of Neurergus crocatus and Neurergus strauchii in Turkey based on morphological and molecular data. Herpetologica 65(3):280-291. Papenfuss T, Sparreboom M, Tok V, Ugurtas IH, Seville M, Kuzmin S, Anderson S, Eken G, Kilic T, Gem E. 2009. Neurergus strauchii. In: IUCN 2011. IUCN Red List of Threatened Species. Version 2011.2. [Online]. Available: http://www.iucnredlist.org [Accessed: 20 May 2012], Pasmans F, Bogaerts S, Woeltjes T, Carranza S. 2006. Biogeography of Neurergus strauchii barani Oz, 1994 and A. s. strauchii (Steindachner, 1887) (Amphibia: Salamandridae) assessed using morphological and molecular data. Amphibia-Reptilia 27(2):281-288. amphibian-reptile-conservation.org 026 September 2012 | Volume 6 | Number 4 | e53 Conservation, husbandry, and captive breeding of the Anatolia newt, Neurergus strauchii Schmidtler JJ, Schmidtler JF. 1970. Morphologie, Biolo- gie und Verwandtschaftsbeziehungen von Neurergus strauchii aus der Turkei. Senckenbergiana Biologica 51:42-53. Schmidtler JF. 1994. Eine Ubersicht neuerer Untersuc- hungen und Beobachtungen an der vorderasiatischen Molchgattung Neurergus Cope, 1862. Abhandlungen und Berichte fur Naturkunde 17:193-198. Schneider C, Schneider W. 2010. Fieldnotes on the ecol- ogy and distribution of Neurergus crocatus Cope, 1862 and Neurergus strauchii strauchii (Steindachner, 1887) in Turkey (Amphibia: Caudata: Salamandri- dae). Herpetozoa 23 (1/2): 59-69. Schultschik G. 2010. (Salamanderland) Neurergus strauchii strauchii (STEINDACHNER, 1887) - Strauchs-Bachsalamander - Strauch’s Brook Salaman- der. [Online]. Available: http://www.salamanderland. at/ Artenliste/N . str. strauchii/NEURERGU S_STR_ STRAUCHII.htm [Accessed: 15 May 2012]. Schultschik G. 2012. Neurergus strauchii (Steindachner, 1887) - Strauchs Bachsalamander. In: Gefahrdete Molch- und Salamanderarten der Welt, Richtlinien fur Erhaltungszuchten. Editors, Schultschik G, Grosse WR. Mertensiella 20. Deutsche Gesellschaft fur Her- petologie und Terrarienkunde (DGHT) e. V., Rhein- bach, Germany. (In press). Sparrboom M, Steinfartz S, Schultschik G. 2000. Court- ship behaviour of Neurergus (Caudata: Salamandri- dae). Amphibia-Reptilia 21(1):1-11. Sparreboom M. 2009. Neurergus strauchii (Stein- dachner, 1887) Anatolia newt. In: Salamanders of the Old World. [Online]. Netherlands Centre for Biodi- versity Naturalis, Leiden, The Netherlands. Available: http://science.naturalis.nl/salamanders [Accessed: 12 June 2012]. Spitzen Van der Sluijs A, Martel A, Wombwel E, Van Rooij P, Zollinger R, Woeltjes T, Rendle M, Haeseb- rouck F, Pasmans F. 2011. Clinically healthy amphib- ians in captive collections and at pet fairs: A reservoir of Batrachochytrium dendrobatidis . Amphibia Rep- tilia 32(3) :4 19-423. Steindachner F. 1887. Uber eine neue Molge-Art und eine Varietat von Homalophis Doriae Pet. Sitzberichte derAkademie kaiserlichen der Wissenschaften (Wien) 96:1-4. Steinfartz S. 1995. Zur Fortpflanzungsbiologie von Neu- rergus crocatus und Neurergus strauchii barani. Sala- mandra 3 1(1): 15-32. Steinfartz S, Hwang UW, Tautz D, Oz M, Veith M. 2002. Molecular phylogeny of the salamandrid genus Neu- rergus: Evidence for an intrageneric switch of repro- ductive biology. Amphibia-Reptilia 23(4):419-431. Steinfartz S, Schutlschik G. 1997. Die Gattung Neurer- gus. Reptilia (Munster) 8:43-48. Stohr A.C, Fleck J, Mutschmann F, Marschang R.E. (In prep.). Ranavirus infection in a group of wild-caught Lake Urmia newts {Neurergus crocatus ) imported into Gennany from Iraq. Received: 14 June 2012 Accepted: 01 July 2012 Published: 22 September 2012 Serge Bogaerts has been fascinated by salamanders and newts since the age of seven. He studied biol- ogy at Raboud University Nijmegen, specializing in herpetology and animal ecology. He works as advi- sor on ecology and nature law for infrastructure projects for the Ministry of Infrastructure and Environ- ment, while he continues herpetological studies as a hobby. Both through field work and captive care, he is striving to leam more about the ecology and behavior of newts and salamanders, particularly those of the Mediterranean and Middle East, and he publishes the results of these studies whenever possible. Henry Janssen started out as a turtle enthusiast in the early 1970s. Gradually his interest shifted to newts, in particular to the genus Paramesotriton. Over the years he was able to build up a significant collection of Paramesotriton and other newt species, and has bred and raised at least one generation of most of these. By keeping detailed records of his observations and through the systematic gathering of data, he has acquired a thorough knowledge about the species he works with. amphibian-reptile-conservation.org 027 September 2012 | Volume 6 | Number 4 | e53 Bogaerts et al. Jennifer Macke has had a life-long fascination with animals. Her interest in newts began with an un- dergraduate research project on limb regeneration, and she has kept and bred caudates ever since. She is particularly interested in newts of the genera Cynops and Neurergus. She is currently employed as a molecular biologist and also volunteers her time to manage the care of the reptiles, amphibians, and invertebrates at a local nature center. Gunter Schultschik started keeping newts and salamanders as a boy and still considers himself just an enthusiast. After getting in contact with AG-Urodela of DGHT in 1989, his collection grew rapidly. J. F. Schmidtler (Munich) was his teacher when he began to travel through Anatolia and the Middle East searching for amphibians and reptiles. As a member of the Austrian Herpetological Society, he founded “Urodela-Austria” a working group which has a meeting once a year, the “Molchlertag.” After some publications together with W. Grosse, they started a project called “Captive Care Management” for threatened species of tailed amphibians, which will be published soon in the DGHT series Mertensiella. Kristina Ernst has been interested in all kinds of animals since she was a small child. Her fascination for newts and salamanders started in 1 993 with her first newts of the genus Cynops. In 2000 she became a member of AG-Urodela of the DGHT and discovered, and developed a special interest in the genus Neurergus. Since then, she has focused on this genus and is responsible for the studbook of Neurergus strauchii at AG-Urodela. Francois Maillet has been keeping and breeding salamanders and newts as long as he can remember. Together with Jean Raffaelli and Amaud Jamin, he forms the core of the French Urodela Group (FUG), whose goals are to bring together knowledge and experience regarding captive bred animals, and to keep stable long-term populations of many species of newts and salamanders in captivity. Christoph Bork has been interested in amphibians since he was six years old. He especially loves newts and salamanders, which he has kept seriously for at least 25 years now. He got his start with Triturus species, and mainly keeps Neurergus species nowadays. As a hobby newt enthusiast he has been a member of the AG Urodela for many years. Additionally he is fascinated by poison dart frogs. Several of these species decorate his living room. amphibian-reptile-conservation.org 028 September 2012 | Volume 6 | Number 4 | e53 Conservation, husbandry, and captive breeding of the Anatolia newt, Neurergus strauchii Frank Pasmans has been fascinated by urodelans since he was a young boy, and he has kept and bred several species. As a veterinarian, he is currently head of a research group that studies amphibian dis- eases at Ghent University, Belgium. Patrick (Pat) Wisniewski (1954-2008) was an all- round natural historian and the longest serving cu- rator of Martin Mere in Lancashire, one of the nine UK Wildfowl & Wetland Trusts centres. He was “the newt man’s newt man,” said to be ahead of his time in amphibian husbandry. He kept and bred a great range of amphibians, especially newts and salamanders — mainly European and Asian species. A vast collection of captive animals covered every inch of several rooms and part of the garden. Pat wrote the booklet “Newts of the British Isles” pub- lished in 1989. amphibian-reptile-conservation.org 029 September 2012 | Volume 6 | Number 4 | e53 Copyright: © 2013 Rastegar-Pouyani et al. This is an open-access article distributed under the terms of the Cre- ative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License, which permits unrestricted use for non-commercial and education purposes only provided the original author and source are credited. Amphibian & Reptile Conservation 6(4):30-35. Ecology of Kurdistan newt ( Neurergus microspilotus : Salamandridae): Population and conservation with an appraisal of the potential impact of urbanization 1>2 Nasrullah Rastegar-Pouyani, 1 2 Mohsen Takesh, 1>2 Akbar Fattahi, 3 Marzieh Sadeghi, 3 Fatemeh Khorshidi, and 4 Robert Browne 1 Department of Biology, Faculty of Science, Rail University, 6714967346 Kertnanshah, IRAN 2 Iranian Plateau Herpetology Research Group (IPHRG), Faculty of Science, Razi University, 6714967346 Kermanshah, IRAN * Faculty of Chemistry, Razi University, Kertnanshah, Iran 4 Royal Zoological Society of Antwerp, Koningin, Astridplein 26, 2018, Antwerp, BELGIUM Abstract. — The Kurdistan newt, Neurergus microspilotus Nesterov, 1916, inhabits springs, ponds, brooks, streams, and wet caves in the western Iranian Plateau in both Iran and Iraq. The Iranian distribution of N. microspilotus is limited to Kurdistan and Kermanshah Provinces. Several major populations of N. microspilotus are threatened by urban development. We gathered autecologi- cal data of N. microspilotus and evaluated factors that may affect the distribution and abundance of this species. We conducted visual surveys for N. microspilotus at twelve localities across the north-western regions of Kermanshah Province from February to July 2012. The survey sites were classified as undeveloped or developed based on their proximity to urban or rural landscapes, and other anthropogenic disturbance and structures. We analyzed the effect of ecological factors, including water pH and specific conductance, temperature, peak of mating behavior, and the time of egg-laying. The daily air temperature of the study sites was provided by the weather bureau of Kermanshah Province. We investigated the correlation between daily maximum air temperature and N. microspilotus population density using Pearson Correlation Analysis, and analyzed the impact of urbanization on specific conductance and pH of habitat water and numbers of N. microspilotus according to Independent-Samples f-test. The densities of N. microspilotus across sites were posi- tively correlated with increased water and daily maximum air temperatures. In addition, we found that densities of N. microspilotus at undeveloped sites were significantly higher than those of de- veloped sites, whereas no relationship was recorded between specific conductance and pH of the water and urbanization. Key words. Kurdistan newt, Neurergus microspilotus, ecology, conservation, urbanization Citation: Rastegar-Pouyani N, Takesh M, Fattahi A, Sadeghi MS, Khorshidi F, Browne R. 2013. Ecology of Kurdistan newt (Neurergus microspilotus: Sal- amandridae): Population and conservation with an appraisal of the potential impact of urbanization. Amphibian & Reptile Conservation 6(4):30-35(e58). Introduction Many species of amphibians globally have declined in abundance and range over recent decades (Collins and Storfer 2003; Stuart and Chanson 2004; Beebee and Griffiths 2005) and 30% of species are now threatened with extinction (IUCN 2010). Related causes of these de- clines and extinctions are habitat loss and fragmentation, unsustainable harvesting, environmental contaminants, increasing UV radiation, climate change, introduced predators, and emerging diseases (Young et al. 2001; Collins and Storfer 2003; Baillie et al. 2004). Urbaniza- tion is a substantial cause of habitat loss and fragmenta- tion (McKinney 2002, 2006). Urbanization is a complex process characterized by increasing in human popula- tion density, which generates significant changes in the chemical, physical, and ecological conditions of affected Correspondence. Email: 1 nasrullah.r@gmail.com areas, and specifically results in the creation of new as- semblages of plants and animals, and possible alteration of the types and frequency of disturbance regimes (Mc- Donnell and Pickett 1993; Kinzig and Grove 2001). Urbanization alters hydrology through water extrac- tion, the construction of impervious surface and increased runoff, increase sedimentation, and pollution of hydro- logical systems (Paul and Meyer 2001; Miltner et al. 2004), and through modifying soils (Effland and Pouyat 1997). Urbanization may also result in an increase in in- vasive plants and animals (Pickett et al. 2001; McKinney 2006), different climates between urban and surrounding rural areas (Grimm et al. 2008). Urbanization is therefore currently one of the most pervasive causes of natural eco- system modification globally, and thus presents a major threat to biodiversity conservation (Czech et al. 2000; Miller and Hobbs 2002). amphibian-reptile-conservation.org 030 March 2013 | Volume 6 | Number 4 | e58 Rastegar-Pouyani et al. Previous investigations indicated that the distribution of the Kurdistan newt, Neurergus microspilotus, is in the mid-Zagros range at the bordering regions of Iran and Iraq (Nesterov 1917; Schmidtler and Schmidtler 1975). Najafimajd and Kaya (2010) reported the first observa- tion of N. microspilotus in west-Azarbaijan, Iran, how- ever, molecular studies were not conducted on these specimens to confirm their claimed status. Major locali- ties of N. microspilotus are found in both urban and rural areas some of which are centers for tourism (Sharifi et al. 2004; Rastegar-Pouyani et al. 2005; Rastegar-Pouyani 2006). In the present study, we investigated the relation- ships between presence and density of N. microspilotus, and the degree of urbanization, and analyzed factors that may affect this relationship including autecological data and water temperature. Materials and methods Study sites and survey techniques The survey area is located in north-western regions of Kermanshah Province, western Iran, and surveys were conducted from February to July 2012. Twelve sites in- cluding a range of ponds, pools, brooks, and streams were selected for surveys. The selection of sites was based on our previous knowledge of these sites providing a consis- tent occurrence of N. microspilotus. Sites investigated were Darian (35°08' N 46° 19' E), Darre-Najjar (35°06' N 46°19' E), Deshe (35°04' N 46°16' E), Dorisan (35°1E N 46°23' E), Hajij (35°09' N 46°19' E), Kavat (34°53' N 46°3E E), Lashgargah (35°01' N 46°08' E), Nilan (35°09' N 46°12' E), Nodeshe (35°1E N 46° 14' E), Noseme (35W N 46°22' E), Qholani (34°54' N 46°27' E), and Qhuri-Qhala (34°2E N 46°30' E) (See Table 1). We categorized study sites into two categories; 1) Developed-sites placed at the center or vicinity of urban, rural, or tourism areas, and 2) Undeveloped-sites remote from urbanization with limited ecological change such as grazing. The counting of N. microspilotus in the Kavat and Dorisan habitats begun on 08 March 2012 and, with weekly intervals, ended on 05 July 2012. Neurergus mi- crospilotus were surveyed and counted through stream bank observation without substrate disturbance. The peak of mating behavior was recorded as the maximum amount of courting behavior, and the time of egg-laying through the observation of eggs in the water for the first time during the season. Collection of habitat data To assay the specific conductance and pH of water, wa- ter-sampling was performed on a 50 ml water sample from each site. A Jenway 3345 Ion Meter was used for determination of conductivity measurements. The pH of water was calculated via pH meter model Metrohm 827 pH lab equipped with a combined glass electrode, calibrated against two standard buffer solutions at pH 4.0 and 7.0 and used for monitoring of the pH values. The daily maximum air temperature of the study sites, over the period of the study, was provided by the weather bu- reau of Kermanshah Province (Table 3). Statistical analysis To statistically analyze the effect of urbanization on spe- cific conductance and pH of water, and the population density of N. microspilotus, we subjected the data to Independent-Sample /-tests. To analyze the relationship between daily maximum air temperature and increas- ing populations of A. microspilotus subjected the data to Table 1 . Study site names and coordinates (North, East), N. microspilotus numbers (no.), water specific conductance (SC; pS cm 1 ), and pH, natural (normal font) or developed (italic font) sites, and threats. Sites Coordinates No. SC pH Threats from development Kavat 34°53'N46°31'E 750 0.3 8.2 — Qholani 34°54' N 46°27' E 79 0.4 7.8 — Darre-Najjar 35°06'N46°19'E 19 0.4 7.6 — Darian 35°08'N46°19'E 24 0.2 7.8 Fish aquaculture and gardening Nilan 35°09'N46°12'E 48 0.3 7.6 Gardeners in this suburban development, clearing the bottom of the pools of aquatic plants that provide shelter for eggs, larvae, juveniles, and adults Hajij 35°09'N46°19'E 7 0.3 7.4 Dam-construction in the Sirvan River, ecotourism, gardening Noseme 35°00' N 46°22' E 5 0.3 7.2 Habitat degradation or loss through irrigation and domestic water usage, accumulation of rubbish in water, and home-construction in the village Deshe 35°04'N46°16'E 1 0.3 7.3 ft Dorisan 35°01'N46°23'E 58 0.3 7.3 ft Nodeshe 35°11' N 46°14' E 30 0.3 7.5 Organic-pollution of water and gardening Qhuri-Qhala 34°21' N 46°30' E 3 0.4 7.4 Major tourist destination. Accumulation of rubbish in the water and the ma- nipulation of habitat through sanitation processes and cleaners in streams Lashkargah 35°01' N 46°08' E 5 0.46 7.6 Many N. microspilotus are road fatalities amphibian-reptile-conservation.org 031 March 2013 | Volume 6 | Number 4 | e58 Ecology of Neurergus microspilotus Table 2. Statistical analysis of the effect of urbanization on the specific conductance and pH of water and the number of N. microspilotus . Variable Developed habitats ( n = 9) Mean ± SD Undeveloped habitats ( n = 3) Mean ± SD t df p-value Specific conductance 0.3 ± 0.1 0.48 ± 0.1 1.1 10 0.32 pH 7.5 ± 0.2 7.9 ± 0.3 2.9 10 0.02 Number 20.1 ± 21.3 299.7 ± 393.9 2.4 10 0.04 Pearson Correlation Analysis using the program SPSS (version 16 for Windows; SPSS Inc. Chicago, Illinois, USA). Data were considered statistically different at P <0.05. Results and discussion for the reproduction of N. microspilotus within the Ka- vat and Dorisan habitats, as well as on Shahoo Mountain (northwestern Kermanshah Province), gradually reaches an optimum with the onset of increased moisture (from melting snow and spring rain) and temperature, with a peak in mid- June and early July (Fig. la, b). Our results suggest that nine out of 12 localities of N. microspilotus from the populations were lowered by ur- banization. These localities are Darian, Deshe, Dorisan, Hajij, Lashgargah, Nilan, Nodeshe, Noseme, and Qhuri- Qhala. The Independent-Samples /-test revealed that devel- oped/undeveloped sites do not have any difference in specific conductance (p = 0.31), but do in the water pH and number of newts (p = 0.04). Bowles et al. (2006) used specific conductance to investigate the effect of urbanization on water of habitats in Eurvcea tonkawae, but our results indicate that specific conductance could not be used as a separator tool to measure the impact of urbanization on N. microspilotus. The resulting data of the specific conductance indicates that there is not much overlap between developed (0.22 < X < 0.46) and un- developed sites (0.27 < X < 0.44). Instead, the analysis suggests that the pH is a better indicator (p = 0.02) of the effects of urbanization on N. microspilotus (in our stud- ied populations) (Table 2). The resulting data on the counting of the newt in the Kavat and Dorisan habitats, the temperature of the two synoptic stations, dates of observations and statistical as- sessment of correlation between maximum daily air tem- perature, and the number of N. microspilotus are present- ed in Table 3. The Pearson Correlation Analysis revealed that there is a strong association between the temperature and presence of individuals of two populations in Kavat and Dorisan (/7-value = 0.919, r = 0.000; /7-value = 0.812, r = 0.000, respectively). According to the data, N. microspilotus adjusts its transition from torpor, and presence in the environment and mating behavior, at a time when food availability of insects and other invertebrates is maximum and the thick- ness of the forest canopy and leaves on the water surface provides the maximum shelter from predators (Table 3; Fig. la, b). At this time a maximal cover of aquatic vegetation provides the best environment for reproduc- tive activities, the deposition of sperm, and egg attach- ment. In March and early April, the vegetative cover of the Kavat and Dorisan habitats is low. Habitat suitability Acknowledgments. — We wish to thank the Mohamed Ben Zayed Foundation, and the European Association of Zoos and Aquaria for supporting of this project through grants. Support was also provided by core funding from the Royal Zoological Society of Antwerp. We also thank the weather bureau of Kermanshah Province, especial- ly Mr. Shaygan for providing weather data, the driver, Sabzali Rasooli, who helped us very much during field work in western Iran, and Mohammad Reza Ashrafi Kooshk and Dariush Naderi for their help in preparation of the manuscript. Literature cited Baillie JEM, Hilton-Taylor C, Stuart SN (Editors). 2004 IUCN Red List of Threatened Species. A Global Spe- cies Assessment. IUCN, Gland, Switzerland and Cam- bridge, United Kingdom. Beebee TJC, Griffiths RA. 2005. The amphibian decline crisis: A watershed for conservation biology? Biologi- cal Conservation 125(2):27 1-285. Bowles BD, Sanders MS, Hansen RS. 2006. Ecology of the Jollyville Plateau salamander ( Eurycea tonkawae'. Plethodontidae) with an assessment of the potential effects of urbanization. Hydrobiologia 553(1): 111- 120 . Collins JP, Storfer A. 2003. Global amphibian declines: sorting the hypotheses. Diversity and Distributions 9(2):89-98. Czech B, Krausman PR, Devers PK. 2000. Economic as- sociations among causes of species endangerment in the United States. BioScience 50(7):593-601. Effland WE, Poutat RV. 1997. The genesis, classification, and mapping of soils in urban areas. Urban Ecosys- tems 1(4): 2 17-228. Grimm NB, Faeth SH, Golubiewski NE, Redman CL, Wu J, Bai X, Bridges JM. 2008. Global change and the ecology of cities. Science 319(5864):756-760. IUCN, Conservation International, and Nature Serve. 2006. Global Amphibian Assessment. [Online] Avail- able: http://www.globalamphibians.org [Accessed: amphibian-reptile-conservation.org 032 March 2013 | Volume 6 | Number 4 | e58 Rastegar-Pouyani et al 0) _3 to > O o o o o o ON o CM oo O 3 in o NO m cc cc C 3 o> CM O in ^1" cn oo in cc o m C 3 CM CM O ON NO CM CM cc m in ON CM C 3 m o ON in ON CM ""t in oo CM C 3 00 o o in »n CM ON ""t CM C 3 O -T CM CM m m CM CM to CM O cc CM m o CM -H O T3 § £ o CD 2 c 3 D Oh CO Q. < h- CM Q. < O CM O m CM O rc O O in ON r- o CM CM CM ON ON CM NO NO CM O CM NO O CM cc CM CM CM OO »n oo CM C3 § 4) -o Co S s CJ o CS -O .a o S-H 4) a § 4> H a* 2 £ Q. < CO Q. < CO to © CO CO CO CM to m to oo ON in in in ON CO o CM 12 a> 0 E ra i_ ra a. re > (0 S2 a> +j o ro c s = J2 3 re 0 re 5 v- s£> VO CN in ■'t Os so oo oo rn oo oo iri C re 'E O a c S2 0 -K o re c s = J 2 3 re o — 0 o 3 +- 2 2 0 !5 a. re E ■= 0 c +* re C re -c 0 o s a ■* — numb ers of n ew t £ t- 5= - b -* — weather temperature 40 35 30 a- B TT 25 mm &.■ :o 15 a- i—i C? 10 4 ■ 0 Figure la, b. 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Population declines and priorities for amphibian con- servation in Latin America. Conservation Biology 15(5): 1213-1223. Received: 22 October 2012 Accepted: 29 December 2012 Published: 14 March 2013 Nasrullah Rastegar-Pouyani earned his B.S. in zoology from Razi University Kermanshah, Iran, in 1986 and his M.S. in zoology from Tehran University, Tehran, Iran, in 1991, where he studied herpetology with the aga- mids as the central object. He started his Ph.D. in Gothenburg University, Sweden, in 1 994 under the advisement I of Professor Goran Nilson and graduated in 1999, working on taxonomy and biogeography of Iranian Plateau agamids, with Trapelus as the main objective. He is a professor and head of the biology department at Razi University. His research interests include taxonomy and biogeography of the Iranian Plateau, the Middle East, ar| d central Asian herpetofauna. Mohsen Takesh earned his B.S. in animal biology from Urmia University, Urmia, Iran, in 2009 and his M.S. in animal biosystematic from Razi University, Kermanshah, Iran, in 2012 under Professors Mozafar Sharifi and Nasrullah Rastegar-Pouyani. His M.S. thesis was on ecology and distribution of Kurdistan newt, Neurergus microspilotus , in its Iranian distribution range. His research interests include ecological, phylogeographical, and phylogenetic investigations of amphibians and reptiles. Akbar Fattahi earned his B.S. from Urmia University, Urmia, Iran, in 1993 and his M.S. in animal biosyste- matic from Razi University, Kermanshah, Iran, in 2012 under the supervision of Professor Nasrullah Rastegar- Pouyani. His M.S. thesis was on ecology and systematics of the Kurdistan newt, Neurergus microspilotus , in its Iranian distribution range and the tree-frog, Hyla savignyi, in Kermanshah Province, western Iran. His research interests include ecological and taxonomic investigations on amphibians and reptiles. amphibian-reptile-conservation.org 034 March 2013 | Volume 6 | Number 4 | e58 Rastegar-Pouyani et al. Marzieh Sadeghi earned her B.S. in applied chemistry from Razi University Kermanshah, Iran, in 2003 and her M.S. in analytical chemistry from Shiraz University, Shiraz, Iran, in 2005, where she studied construction of optical sensor for pH and metal ions under the advisement of Professor Afsaneh Safavi. She started her Ph.D. in Razi University, Kermanshah, Iran, in 2005 under the advisement of Professor Mojtaba Shamsipur and gradu- ated in 2010, working on optical and electrochemical sensor for metal ions and pharmaceutical compounds as the main object. Her research interests include optical sensor and liquid phase microextraction, solid phase extraction, and removal of toxic compound from aqueous environment using nanomaterial compound. Fatemeh Khorshidi earned her B.S. in applied chemistry from Razi University Kermanshah, Iran, 2006 and her M.S. in analytical chemistry from Razi University, Kermanshah, Iran, in 2012, where she studied the removal of arsenic from aqueous environment using metallic nanocomposite under the advisement of Dr. Marzieh Sadeghi. Her research interests include removal of toxic elements from aqueous environments, nano-drug and medical nano technology. Robert Browne is co-editor of the journal Amphibian and Reptile Conservation. He has a wide range of aca- demic and practical experience in many research fields supporting herpetological conservation and environmen- tal sustainability. Currently he works for “Sustainability America” www.SustainabilityAmerica.org (Belize). amphibian-reptile-conservation.org 035 March 2013 | Volume 6 | Number 4 | e58 Copyright: © 2013 Najafi-Majd and Kaya. This is an open-access article distributed under the terms of the Cre- ative Commons Attribution-NonConunercial-NoDerivs 3.0 Unported License, which permits unrestricted use for non-commercial and education purposes only provided the original author and source are credited. Amphibian & Reptile Conservation 6(4):36-41. Rediscovery of the Lake Urmia newt, Neurergus crocatus Cope, 1862 (Caudata: Salamandridae) in northwestern Iran after 1 50 years ^Inaz Najafi-Majd and 2 Ugur Kaya Ege University, Faculty of Science, Department of Biology’, Section of Zoology, Bornova, izmir/ 35100, TURKEY Abstract . — We report on the rediscovery of the Lake Urmia newt, Neurergus crocatus in Iran, 150 years after its original description and last report by Cope 1862. The Lake Urmia newt is classified as Vulnerable by the IUCN Red List of Threatened Species. Some specimens, both adult and larvae, of N. crocatus were found in Iran during two field surveys near the Iran-lraq border (south west of West Azerbaijan Province, surrounding the type locality “Urmia,” at 1786-1823 m above sea level [a.s.l.] elevation). Water samples were taken from two breeding habitats, as preliminary data, and were ana- lyzed for 13 chemical variables to determine the characteristics of water chemistry. The morphologi- cal comparison of the new specimens with the original description and data from Schmidtler and Schmidtler (1975) did not reveal any distinct morphological differences. Previous to our study there was no information regarding the exact locality of N. crocatus and its population status in Iran. Our confirmation of N. crocatus in northwestern Iran indicates that protection is needed if this species is to persist in Iran. In addition, water chemistry analysis of the two new habitat records showed that in this area N. crocatus inhabits two streams with good water quality. Key words. Neurergus crocatus , rediscovery, conservation, Iran Citation: Najafi-Majd E, Kaya U. 2013. Rediscovery of the Lake Urmia newt, Neurergus crocatus Cope, 1862 (Caudata: Salamandridae) in northwestern Iran after 150 years. Amphibian & Reptile Conservation 6(4):36-41(e59). Introduction Newts of the genus Neurergus (Salamandridae) are con- fined to Turkey, Iran, and Iraq. Neurergus was originally categorized as a member of the family Salamandridae and the subfamily Tritoninae, subsequently it was changed to Pleurodelinae (Cope, 1862). Four species of Neurer- gus have been described (Schmidtler 1975; Leviton et al. 1992; Sparreboom et al. 2000): Neurergus crocatus Cope, 1862 from northwest of Iran, northern Iraq, and southeast of Turkey; Neurergus strauchii (Steindacliner 1887) from the western side of Van Lake to Malatya in eastern (Anatolian) Turkey; Neurergus microspilotus (Nesterov 1916) from the west and northwest of Iran and east of Iraq; and Neurergus kaiseri Schmidt 1952 from the surroundings of Shah-Bazan of Luristan Province, Iran. All known species of Neurergus can easily be dis- tinguished by their morphological characters (Schmidtler and Schmidtler 1970, 1975; Schmidtler 1994; Najafimajd and Kaya 2010; Schneider and Schneider 2011). At pres- ent, the taxonomic relations of the closely related taxa N. microspilotus and A. derjugini (Nesterov 1916) from the Iraq and Iran borders are still debatable (Schneider and Schneider 2011). The genus Neurergus is represented by three species in Iran, N. crocatus Cope, 1862, N. microspilotus (Nest- erov 1916), and N. kaiseri Schmidt, 1952 (Balouch and Kami 1995). According to IUCN, N. crocatus has one of the largest distribution ranges among all Neurergus spe- cies; though there is almost no data about its biology and exact distribution (Steinfartz et al. 2008). As stated by Sparreboom (2009) this insufficient data may be ascribed primarily to geographic inaccessibility, as well as long term ethnic tensions, and a long and continuing history of military conflicts. Neurergus crocatus has been known from several localities in Turkey since 1986 (Baran and Oz 1986; Baran and Atatur 1997; Ozdemir et al. 2009). This striking coloration of these N. crocatus attracted the attention of local people and non-herpetologists subse- quently misidentified them as N. strauchii (Kemal 2008). Very recently in 2010 a new locality for N. crocatus, in- cluding two neighboring streams close to the §emdinli, Turkey, was published (Schneider and Schneider 2010). Neurergus crocatus has been reported from eight lo- calities in Iraq: Agrah (Schmidt 1939), Shiwolak and Tajeka villages in the northeastern (Khalaf 1961; Nader 1969), and five recent localities from Barzan, Girbish, Roste, Smilan, and Nawanda in 2010 (Schneider and Schneider 2011). Correspondence. Email: 2 ugurkaya@ege. edu.tr (corresponding author); 1 elnaz.najafi.majd@gmail.com amphibian-reptile-conservation.org 036 April 2013 | Volume 6 | Number 4 | e59 Najafi-Majd and Kaya There is limited information regarding the exact local- ity and the distribution of N. crocatus and its population status in Iran. Only one historic record by Cope (1862) is noted from northwestern Iran but the exact type locality is unknown, however Fowler and Dunn (1917) reported the type locality as “Ooremiah, Persia.” Within the Salamandridae, newts of the genus Neu- rergus are basically known as stream inhabiting species that leave streams during dry periods or during winter (Schmidtler and Schmidtler 1970, 1975; Schmidtler 1994). The Lake Urmia newt, N. crocatus is quite close- ly related in an ecological aspect to Calotriton species which have similar habitats (Steinfartz et al. 2002). Neu- rergus crocatus is a montane species and lives in cool and well-oxygenated streams (Ozeti and Yilmaz 1994; Baran and Atatiir 1998), where it breeds. There is no published information about the terrestrial component of their life history. It is presumed that the adults hibernate under rocks and other cover during the winter (Papen- fuss et al. 2009). There is almost no information on its life cycle, but eggs and larvae of different lengths were observed at the end of May and June. The seasonality of the breeding season is dependent upon elevation (Ozeti and Yilmaz 1994). According to the IUCN, there is a continuing decline in the extent and quality of the habitat of N. crocatus in Turkey, Iran, and Iraq (Papenfuss et al. 2009). However, there is no published information about the size and rate of this decline. Neurergus crocatus is categorized as a Vulnerable species [VU B2ab (iii)] in the IUCN Red List of Threatened Animals. Despite the significance of its conservation, nothing is known about the life history of N. crocatus , e.g., individual growth, longevity, and other demographic parameters, or its ecology including critical habitat components such as water quality and tempera- ture, breeding, oviposition, or hibernation sites. One hundred and fifty years after the original descrip- tion of N. crocatus (Cope 1 862) we verify the existence of N. crocatus in northwest Iran. We also provide infor- mation on the characteristics of water chemistry in the aquatic breeding habitat of N. crocatus. Materials and methods To assess the presence of N. crocatus in north-west Iran, we conducted two surveys during the field seasons of 2009 and 2010 of the northern Zagros Mountains in the west of Lake Urmia, from Sero (the area between Lake Table 1 . Morphological characteristics of the closely related species Neurergus microspilotus and N. crocatus (Schmidtler and Schmidtler 1975) in comparison with our specimens from Oshnaviyeh. Characteristics Neurergus crocatus Neurergus microspilotus Specimens from Oshnayiyeh Adult 1. Maximum total length of $/c? 16 / 18 cm 15 / 17 cm 16.8 cm $ (Mean) 2. The form of cloacae in mating season in $ Lips approximately 1-2 mm protruding Not sharp conical, lips 3 mm protruding Lips approximately 1-2 mm pro- truding 3. Design of backside (spots small 0.2 mm, large 0.4 mm) Large and small yellow spots Small yellow spots Large and small yellow spots 4. Design of throat Unicolored orange Orange, mostly with black spots Unicolored orange 5.Design of belly Unicolored yellow to reddish orange Lateral black coloration con- fines orange middle parts Unicolored yellow to reddish orange 6. Design of underside of extremities Unicolored orange Orange, mostly black spotted Unicolored orange 7. Design of tail laterals Large yellow spots Small yellow spots Large yellow spots Larvae 8. Total length 35-70 mm 35-70 mm 58 mm 9. Relation dorsal fin- 1 . 0 - 1.1 Protrudes from the back center significantly 0.7-0. 9 1 . 0 - 1. 1 Protrudes from the back center significantly length / interaxial length Protrudes from the back center 10. Dorsal design of older larvae (about 50 mm total length) Bright spots irregularly and big; partly fussed Clearly long bright stains toward two lines along center of the back Big, irregular bright spots; partly fussed 1 1 . Design of belly of older larvae (about 50 mm total length) Almost unicolored bright Tow imperfect dark spot lines along edges of belly Almost unicolored bright 12. Design of tail sides More or less clearly dark pig- mented Clearly blackish brown "clouded" More or less clearly dark pigmented amphibian-reptile-conservation.org 037 April 2013 | Volume 6 | Number 4 | e59 Rediscovery of the Lake Urmia newt Urmia and Iran-Turkey border) and continued the search to the south up to Piranshahr, in the West Azerbaijan Province near the Iran-Iraq border (between 37° 42’ 36” and 36° 40’ 33” latitudes). We investigated all potential and accessible habitats of N. crocatus in this area, including streams, springs, and ponds. Searches were undertaken between 9.30 am and 6.30 pm. Local people were interviewed in order to obtain more information on N. crocatus. Geographic positions of study sites were recorded with a GPS receiver (Garmin eTrex® 30). Total lengths of adult females were measured with calipers in the field and given in millimeters (mm). Collected specimens were compared morphologically with the description given in Cope (1863) and Schmidtler and Schmidtler’s (1975) table (Table 1). Water samples were collected from the breeding habi- tat to determine parameters of the salamander’s breeding water conditions. Water conductivity, pH, dissolved oxy- gen (DO), and salinity were measured in the field using a Hach Portable pH/conductivity/dissolved oxygen meter, and water temperature was measured with a thermome- ter. Some water chemistry parameters such as iron, man- ganese, chloride, ammonium, sulfide, potassium, nitrate, ammonia, and hardness (calcium and magnesium) were measured in the laboratory using a DR 2800 VIS Spec- trophotometer, following the manufacturer’s procedures. Results and discussion There was no evidence of the species hi the 2009 field survey but in 2010, on June 4 th , seven adult specimens (?) were discovered and collected from a Margo Ziyarat Region spring near Oshnaviye, at the border in the west of West Azerbaijan Province, north-Zagros Mountains. In the last conducted survey of the same locality on July 30, 2010 morning, two adults in the spring and eight lar- vae were found in a small stream and six larvae were collected from the stream; elevation of the location was 1786-1823 ma.s.l. Neurergus crocatus was previously known from 11 locations in Iran, Iraq, and Turkey (Cope 1863; Schmidt 1939; Khalaf 1961; Nader 1969; Baran and Oz 1986; Schneider and Schneider 2010, 2011). This species is present in the vicinity of Beytu§§ebap and Semdinli, south-east Anatolia, Turkey and in eight localities in the northeastern region of northern Iraq. For the Iranian re- cord, Cope (1862) did not designate the exact type lo- cality of N. crocatus ; however subsequently Flower and Dunn (1917) determined the type locality as “Ooremi- ah, Persia” which corresponds to Lake Urmia. Freytag (1956: pi. 4) has at first depicted the type specimen with the labels. We discovered N. crocatus in the west of West Azerbaijan Province near the Iran-Iraq border in a natural spring and a small stream. The investigated locality and previously known localities are shown in Fig. 1. In the north west of Iran in the Mergo Ziyarat region near Oshnaviyeh, we found a total of 17 newts (nine adult females and eight larvae); adults from a spring and larvae from a stream. Adult specimens were discovered in Arabe Spring (37° 2.59’ N; 44° 56.72’ E), west of Os- hnaviyeh. Six larvae were collected from Gurgu Stream (37° 2.78’ N, 44° 56.80’ E), approximately 100 m north of the spring at 1786-1799 m a.s.l. (Fig 2: A, B). Females were hiding between vegetation in the spring and larvae were found in streams with fast running water, in a small puddle behind stones. Morphological characters Collected specimens were compared morphologically as well as in coloration with the description of Cope 1862 and Table 1 in Schmidtler and Schmidtler (1975). Neu- rergus crocatus is characterized and readily identified by yellowish color spots on their flattened black body. Dor- sal blotches are yellow and small in midline and larger with light yellow coloration in the lateral position; ven- tral surface orange-red in males, yellowish in females; limbs overlap when laid against the body, broadly. Males have white colored spots along the tail (Schneider and Schneider 2010). Tail fins on dorsal and ventral sides de- veloped; dorsal tail fin a little higher, especially in the breeding season (Fig 2: C, D). Our specimens represent the typical characteristics of N. crocatus given in the literature (Table 1 ; Schmidtler and Schmidtler 1975). The species dorsal coloration varies from dark brown to black with numerous yellow round or elongated spots. The yellow spots were a little smaller than in the type specimen (Freytag 1956: pi. 4). The coloration of trunk and tail venter is yellowish or- ange with infrequent small black spots overlaying the abdomen. Ecological habitat characteristics Generally, salamanders are found only in or near running water such as mountain brooks and streams and their sur- vival and distribution can be strongly limited by water quality (Saynn et al. 2009). Moreover, salamanders have highly permeable skin and their larvae may be particu- larly susceptible to water quality parameters (Duellman and Trueb 1994). Some chemicals such as nitrates and chlorides could influence salamander distribution, abun- dance, and the selection of breeding sites (Odum and Zippel 2008). Neurergus crocatus is a montane species that lives and breeds in cool and continuously flowing streams and springs in hillsides and mountainous areas. Habitat substrate is composed of small stones and sands. Typical of the environment of N. crocatus are Water cress (Nas- turtium officinale), Oregano ( Origanum vulgare), and amphibian-reptile-conservation.org 038 April 2013 | Volume 6 | Number 4 | e59 Najafi-Majd and Kaya Figure 1. Anew locality, Oshnaviyeh (11) and other known localities of Neurergus crocatus : 1) Beytii^ebap, 2) Semdinli, 3) Agrah, 4) Shiwolak, 5) Tajeka, 6) Barzan, 7) Girbish, 8) Roste, 9) Smilan, 10) Nawanda. The hatched part shows our studied area, which also covers the unknown exact location of Cope’s original “terra typica.” Figure 2. A) Arabe Spring. B) Gurgu Stream. C and D) Neurergus crocatus adult female. amphibian-reptile-conservation.org 039 April 2013 | Volume 6 | Number 4 | e59 Rediscovery of the Lake Urmia newt Table 2. Values of some water chemistry variables in breeding habitats of Neurergus croccitus. Habitat TEM pH DO CON SAL Fe Mn Cl K NO, NH, Hardness Hardness (C°) mg/I Ms/cm % mg/I mg/I mg/I mg/I mg/l mg/i Mg(mg/I) Ca (mg/l) Arabeh 10 7.69 6.78 228 0.11 0.03 0.2 0.9 0.7 1.2 0.01 2.43 0.01 Gurgu 15 8.45 7.64 301 0.14 0.00 0.2 0.9 0.5 1.2 0.02 2.00 0.13 Stinging nettle ( Urticci dioica ). According to Baran and Atatiir (1998) N. crocatus spend the winter months on land under stones or in burrows. Analysis results of some water chemistry parameters in breeding habitats such as iron, manganese, chloride, potassium, nitrate, ammonia, and hardness (calcium and magnesium) are given in Table 2. Mean values of some water chemistry parameters of breeding habitats was found to be as follows: Fe = 0.015 mg/1, Mn = 0.2 mg/1, Cl- = 0.9 mg/1, K = 0.6 mg/L, NCk = 1.2 mg/L, NFL, = 0.015 mg/L, hardness Ca = 0.07 mg/L, and hardness Mg = 2.215 mg/L. Therefore, in these in- habited waters toxic parameters (chloride, nitrate, and ammonia), hardness (Mg and Ca), dissolved oxygen, and pH are all in acceptable range (Odum and Zippel 2008). Amphibians, especially salamanders, are excellent indicators of local conditions because they have perme- able skins and fairly limited home ranges (Blaustein and Wake 1995). After obtaining similar information from known breeding habitats and comparing these with habi- tats that are not used for breeding, it will be possible to assess the water quality requirements for breeding of N. crocatus. This information can be combined with infor- mation of other habitat variables to assess the broader habitat requirements of N. crocatus. Threats and conservation of N. crocatus Currently, N. crocatus is considered Vulnerable by the IUCN Red List due to its restricted range and potential habitat destruction. Neurergus crocatus has not been ob- served since its original description from Iran by Cope in 1862. Our new record verifies its existence after 150 years from its original description and adds to the broad- er distributional knowledge of the species. The species is susceptible to habitat change, habitat loses, pollution, drought, and over harvesting (Papenfuss et al. 2009), but these factors have not been determined. There is a continuing decline in the extent and quality of its habitat. The distribution of this species in Turkey is expected to undergo significant change due to various human activities such as the construction of several dams within the range of N. crocatus over the next 10 years (Papenfuss et al. 2009). During summer and fall over grazing and pollution of streams by sheep and goats dam- age the habitat of N. crocatus. Some N. crocatus were reported killed by local people that use these springs as drinking water resource, as they think these newts are poisonous. To assure the sustainable management of N. crocatus it is imperative to assess its habitat needs and conser- vation status over its range and distribution surveys. Of particular importance are how aquatic habitat variables influence the choice and success of breeding sites. Acknowledgments. — This study was a part of the first author’s Ph.D. thesis. We want to thank Jahanbakhsh Najafi-Majd who companies us in all field studies. This work was supported by Ege University (BAP, Project No. 2012/FEN/004). Literature cited Baloutch M, Kami HG. 1995. Amphibians of Iran. Teh- ran University Publications 2250, Tehran, Iran. 91-98 (177 p.). Baran I, Atatiir MK. 1998. Turkish herpetofauna (am- phibians and reptiles). Republic of Turkey, Ministry of Environment, Ankara, Turkey 16 (214 p.). Baran I, Oz M. 1986. On the occurrence of Neurergus crocatus and Neurergus strauchii in Southeast Ana- tolia. Zoology in the Middle East (Heidelberg) 1:96- 104. Biricik M. 2009. The re-determination of the Neurergus (Salamandridae, Caudata) specimens recently record- ed in $irvan. Cesa News (Centre for Entomological Studies, Ankara) 46(1 901): 1-5, 6-10. Blaustein AR, Wake DB. 1995. The puzzle of declin- ing amphibian populations. Scientific American 272(4):52-57. Cope ED. 1862. Notes upon some Reptiles of the Old World. 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Phylog- eny of Neurergus crocatus and Neurergus strauchii in Turkey based on morphological and molecular data. Herpetologica 65(3):280-291. Ozeti N, Yilmaz I. 1994. Tiirkiye amphibileri. Ege Uni- versitesi Fen Fakiiltesi Kitaplar Serisi. 22 lp. Papenfuss T, Sparreboom M, Ugurtas I, Rastegar-Pouy- ani N, Kuzmin S, Anderson S, Eken G, KiliQ T, Gem E, Kaya U. 2009. Neurergus crocatus. In: IUCN Red List of Threatened Species. Version 2011.2. [Online]. Available: www.iucnredlist.org [Accessed: 09 April 2012 ]. Saynn F, Ba§kale E, Tarkhnishvili D, Kaya U. 2009. Some water chemistry parameters of breeding habi- tats of the Caucasian salamander, Mertensiella cauca- sica in the Western Lesser Caucasus. Comptes Rendus Biologies 332:464-469. Schmidt KP. 1939. Reptiles and amphibians from south- western Asia. Field Museum of Natural History, Zoo- logical Series 24:49-92. Schmidtler JF. 1994. Eine Ubersicht neuerer Untersuc- hungen und Beobachtungen an der vorderasiatischen Molchgattung Neurergus. 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Molecular phylogeny of the salamandrid ge- nus Neurergus : evidence for an intrageneric switch of reproductive biology. Amphibia-Reptilia (Leiden) 23(4):4 19-431. Rastegar-Pouyani N. 2006. Conservation and distribu- tion of Neurergus microspilotus (Caudata: Salaman- dridae) in Zagros Mountains, Kermanshah Province, western Iran. 13th Congress of the Societas Europaea Herpetologica. 115-116. Received: 19 October 2012 Accepted: 26 December 2012 Published: 05 April 2013 Elnaz Najafi-Majd is originally from the Islamic Republic of Iran. She is currently a Ph.D. student at the Zo- ology Section, Biology Department at Ege University in Turkey. She focuses her studies on the conservation, ecology, and distribution of the Lake Urmia newt (Azerbayjan newt; Neurergus crocatus) and the Kurdistan newt ( Neurergus microspilotus). Ugur Kaya is professor at Ege University and curator of the zoology department Ege University (ZDEU) col- lection. His main research fields are taxonomy, ecology, and bioacoustics behavior of amphibians. Recently, his main research interests have focused on the conservation and ecology of amphibians. To help protect amphib- ians species, he monitors various endangered populations and also uses bioacoustics to monitor other anuran populations. amphibian-reptile-conservation.org 041 April 2013 | Volume 6 | Number 4 | e59 Copyright: © 2013 Al-Sheikhly et al. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License, which permits unrestricted use for non- commercial and education purposes only provided the original author and source are credited. Amphibian and Reptile Conservation 6(4): 42-48. New localities of the Kurdistan newt Neurergus microspilotus and Lake Urmia newt Neurergus crocatus (Caudata: Salamandridae) in Iraq ^mar Fadhil Al-Sheikhly, 2 lyad A. Nader, 3 Nasrullah Rastegar-Pouyani, and 4 Robert K. Browne 1 Omar Fadhil Al-Sheikhly, Department of Biology’ - University of Baghdad, IRAQ 2 P.O. Box 2491, Riyadh, SAUDI ARABIA D Faculty of Science, Razi University, 67 14967346 Kermanshah, IRAN 4 Royal Zoological Society of Antwerp, Antwerp, BELGU for Sarteneja, BELIZE Ipf Biolog y, inability Abstract. — Little is known about the distribution and current conservationftatul^rfthe two M pe- des of mountain dwelling newts of the genus Neurergus found in the Zacu4s Mounn^in nopnern and northeastern Iraq: the Critically Endangered Kurdistan newt Neurerc s micrr oilotu (Nesterov 1916), and the vulnerable Lake Urmia newt Neurergus crocatus (Cop^u62)^Surveys irvlrie Kurd- istan region of northeastern Iraq from 2007 to 2012 resulted in the liscc rj Jf N. m^ospilotus at seven new localities distributed in the Zagros Mountain of Sulayjj^miya Pr^whpe. "Hie new locations provide a major range extension of N. microspilotus. In additipn, tour new I^^M^ of N. crocatus were located between 2007 and 2013. In addition to Neurergus ne^keurveys, imerviews with local people were also conducted through the use of photographs. Severnkought during recent years as well as anthropogenic habitat destruction and pollwc^rcive been considered as main threats to the survival of both species in northern Iraq. Here twaescribe ngw geographical distributions and the conservation status of both Neurergus species Bund in Iraq: Key words. Kurdistan newt, Lake Uremia newt, Nmner gus micl tan region, Iraq Iptus^xeu rergus crocatus , salamanders, Kurdis- Citation: Al-Sheikhly OF, Nader IA, Rastegar-Pouyani N, Browne RK. ^13. newt Neurergus crocatus (Caudata: Salamandridae) if^^^Amphibil lo c a rngp of the Kurdistan newt Neurergus microspilotus and Lake Urmia Reptile Conservation 6(4): 42-48 (e68). Introduction The Kurdistan newts we sun description by Nesterov ( ] triton derjugini and R.. ons, Neurergus d. clerjTtgini (1 ;d cor^Bfided to then under the^knes Rhithro- (crospilotu^mhQSQ tax- ov 1916) and N. d. microspilotus (NjBProv 1916), ar^™rently considered a single speci^Pyn. Nm rergus mi^^pilotus [Nester- ov 1916]) a^^vere ^^^recorded in Iraq by Schneider and Schneide^k&ll), in the d^e vicinity of their type 1 ocaltfBB^I ram^Ee sur^pFmade by Schneider and Sd^lider^k 11) irn^ysBmstan region of northern Iraq _reportecWe Critically Endangered (IUCN icrospilotus from seven localities situa|Bcrm the northeastern mountains along the Iraq- Iran bnkr. Leviton et al. (1992) described the range of the Vuln^^le (IUCN Red List 2013) Lake Urmia newt (TV crocatus ; Cope 1862) from northeastern Iraq, eastern Turkey, northwestern Iran, and the Zagros Mountains of Luristan. Neurergus crocatus was found in Iraq by Allouse (1955) and Khalaf (1959), with a subsequent review of the range and distribution of N crocatus in Iraq by Nader (1969). Al-Adhami and Hameed (1988) carried out a comprehensive study on the histology of the TV. c. cro- catus (Schmidt 1939) combined with description of the sampling locations. Mahdi and George (1969) listed both TV. microspilotus and TV. crocatus in the herpetofauna of Iraq without providing their range or distribution. Both species were shown to have a restricted range in Iraq and to be allopatric (Najafimajd and Kaya 2010). From 2007 to 2013 we conducted intensive field work to de- termine the range, distribution, and conservation status of Neurergus newts in the Kurdistan region of northern Iraq. Here we describe a new geographical distribution for both species in Iraq and assesses their conservation status. Material and Methods We surveyed suitable habitats from 2006 to 2012 for Neu- rergus in the three Iraqi provinces of Kurdistan region in Northern Iraq (Sulaymaniyah, Erbil, and Duhok provinc- es; Table 1); additional field observations were made dur- ing a short survey in 2013. In mountainous landscapes (elevation of ca. 1200-1 600 m) consisting of fresh water springs, streams, ponds, and waterfalls, we conducted surveys in order to locate Neurergus eggs, larvae, and Correspondence. Email: 1 alsheikhlyomar@gmail.com (corresponding author). amphibian-reptile-conservation.org 042 July 201 3 | Volume 6 | Number 4 | e68 Al-Sheikhly et al. Barzan Smilan A ( Nawanda A Shiwalok l Halsho o Awdalan JJ Sulav Ashawa Duhok Iran Smaquli Area W < t « i Erbil *- N. microspilotus (2007-2013) 0 N. crocatus (2007-2013) / N. microspilotus Schneider & Schneider (2011) A* crocatus Schneider & Schneider V 1111 V’^Sulaymaniyah t Ahmad AwaX Sargate Fig. 1 . Google Earth map of northern ^^LtKurdi 2013 for Neurergns microspilotus^^ sll^Vs new localities we discovered during our surveys from 2007 to scribed by Schneider and Schneider (2011). adults. Adults and larvaey^re photogra^^toi and some collected for morphoh^^u^kmnination; e^wfially A. crocatus , with focu^olrthe dimfc^es between the col- oration pattern A. derjugin^^dcrospilotus taxon and those of N^^catus Canon E^^Eanon EOS 40D camera bod^EquipP^P^ith Canon EF 75-300 mm (f 4-5.6) and cSfc^^Ro-400jgjpi (f 4) lens was used to take onl^t to con»n field identifications. We in eir^M^a^^roof Hiking GPS device to >ns (longl^e, latitude, and elevation). Lo- dewed and shown photographs in orcn^^further identify locations where Neurergus spp. have observed. An IUCN threat assessment score was then^pculated to identify the main threats to both Neurergus spp. Results Kurdistan newt (Neurergus microspilotus) Neurergus microspilotus is mainly found at an eleva- tion of about 1200-1600 m in the fresh water springs, streams, ponds, and waterfalls of the Zagros Mountain Forest Steppe Ecoregion in northeastern Iraq. These water bodies are primarily found on hillsides or in deep wooded valleys. The 2012 surveys located populations of A. microspilotus (18 individuals: 12 males; 6 females; 442 larvae) at seven new locations along the Iraq-Iran northern border (Table 1, Fig. 1). The previously recog- nized populations of N microspilotus in Iraq are mainly restricted to the mountains of northeastern districts of Sulaymaniyah province. We found six new localities for N microspilotus in the Halabja and Pshdar districts of Sulaymaniyah province. On 10 July 2007 an adult male N microspilotus was found in a shallow mountain pond with a maximum depth of 7.3 cm. The pond (elevation 1307 m) branched from a running stream within a valley near Isawa village, Mawat Mountain, Sharbazher district (to the north of Su- laymaniyah City). Furthermore, additional new localities for N microspilotus were discovered during extensive field surveys in northern Iraq (in Iraqi Kurdistan) per- formed during May and June 2012. At the Isawa site on 13 May 2012 we failed to locate any adult A. microspilo- tus but found hundreds of eggs and larvae in early meta- morphosis stages. We also surveyed many suitable habi- tats in the Pshdar district on 15 May 2012 which resulted amphibian-reptile-conservation.org 043 July 201 3 | Volume 6 | Number 4 | e68 New localities of the Kurdistan and Lake Urmia newts Table 1. List of the New localities for N. microspilotus and N. crocatus in Northern Iraq (Kurdistan Region); M = male; F =female; L =larva; *= unknown count. Province District Site Coordinates Neurergus species M Sulaymaniya Sharbazher Mawat - Isawa village Sulaymaniya Halabja Sargate Sulaymaniya Halabja Ahmad Awa Area Sulaymaniya Halabja Byara Sulaymaniya Pshdar Qara and Abubakra Area Sulaymaniya Pshdar Halsho Sulaymaniya Pshdar Hero N35°56'E45°23' N35°17'E46° 6' N35°18'E46° 5' N35°13'E46° 7 N 36°24' E 45° 3' N36°12'E45°16' N 36° 7E45°17 microspilotus microspilotus microspilotus microspilotus microspilotus microspilotus microspilotus Total Erbil Shaqlawa Dob Smaquli Area N36°21'E44°19' crocatus Erbil Choman Grtk N 36°46’ E 44°52' crocatus Duhok Duhok Area Ashawa - Sarsank N37°0'E43°17' crocatA Duhok Amedi Sulav N 37° 5’ E 43°27' cv^mtus Total in three new locations for N. microspilotus. Adult males and females were located in a small gravel pool branch- ing from a running mountain stream at elevation ca. 1309 m in the Hero area (to the southeast of the Qaladza town- ship). Searches along the edge of the pool resulted in identifying 17 larvae in different metamorphosis stages. Both adults and larvae were carefully examined andjaho tographed (Figs. 2a and b). We located a breeding site for N. microspilotus elevation of ca.1342 m in a mountain poncHo the of Hero in the Halsho area to the nordB^^BQalad' (ca. 10 km). A total of 23 larvae wem^Rind buBo were found (Fig. 3a). On 9 Jun^E012 t AbubakrJBfeas ^Bkyisited. ATfbr interviewing many Wak wp wprp able^klocate adult males and females w^n ar vae m cimerent metamorphic stages in a all mountainstream at an elevation of ca. 1300 m in Qara mounBn (Fig. 3b). In June 2012, our surveys eurergus Bre extended throughout suitable habi- tatS^^neHjBja district. In addition to the Schneider Scnneiaier (2011) sites, we located three new locali- r . microspilotus in Halabja. dult male along with 3 1 larvae were found in a mountain pond (elevation ca. 1400 m) in Ahmad Awa on 4 June 2012. Additionally, 11 individuals (seven males and four females) with 208 larvae in early metamorphic * M V, >*l’ Figs. 2a and 2b. Neurergus microspilotus (a): adult male; (b): larva, Hero of Pshdar district. Photographs by Omar Al-Sheikhly. amphibian-reptile-conservation.org 044 July 201 3 | Volume 6 | Number 4 | e68 Al-Sheikhly et al. stages were found in Byara (ca. 12 km) to the southeast of Ahmad Awa and near the Schneider and Schneider (2011) sites in Tawale and Balkha (Fig.l). On the 5 June 2012, through interviews with local people, an adult male was located in a mountain stream at an elevation of ca.1254 m in Sargate area in Halabja (Table 1). Lake Uremia newt (Neurergus crocatus) Neurergus crocatus thrives in any suitable aquatic habitat found at an elevation of ca. 1200-1500 m in the north- western parts of the Zagros Mountain Ecoregion in northern Iraq. In addition to the Schneider and Schnei- der (2011) N crocatus localities, our 2012-2013 surveys resulted in four new localities for N crocatus within the Erbil and Duhok provinces. Four males and two females were found (Table 1). However, it appears that there are many potential areas of suitable habitat for N crocatus in northern Iraq still to be discovered. Four new localities (two in Erbil and two in Duhok) for N. crocatus were located during 2007 field surveys, and during a short visit to the Duhok area in 2013 (Fig.l). In the Erbil Province two new localities were located. On 17 August 2007 an adult male and female were found in a mountain stream at elevation of ca. 1400 m in the Grtk Mountains in the Choman district of the Erbil Prov- ince (c. 15 km), close to the Iraq-Iran border (Fig. 4). On 25 August 2007 an adult male was found at elevation of ca. 1200 m in a mountain stream in Doli Smaquli area of Shaqlawa district in Erbil. On 1 September 2007 an adult male was located at the Sulav area i^Aniedi district at elevation of ca. 1400-1500 m. OnJ^^^B®Q13 and during a short visit to the Ashawa S arsank^^erfal 1 an adult male and female was found at an elevatio®f ca. 1206 m (Table 1). Discussion There is a newts in and scatte' tat of info Ferning Neurergus newtSMF^e a restricted range |on mainly confined to the habi- ivtou^un Forest Steep Ecoregion. Our surveys suggest that Neurergus newts are /tS i Figs. 3a and 3b. Neurergus microspilotus (a): juvenile at Halsho of Pshdar district; (b): Adult male (below) and female (above) in Qara mountain. Photographs by Omar Al-Sheikhly. amphibian-reptile-conservation.org 045 July 201 3 | Volume 6 | Number 4 | e68 New localities of the Kurdistan and Lake Urmia newts Fig. 4. Male (right) and female Sheikhly. still thriving in suitatfl^nabita^m tne nortnwn moun- tains of Iraq, witl^^ations con^fctated mainly along the border witl^Rm. Our surveys abided the known range of N. j^^ospilo^fthat included seven new loca- tions in the nkmtau^^n northeastern Iraq. In addition, four neaJocalmBff^ N. crocjrs were discovered in this in mountain stream at Grtk of Choman district in Erbil. Photograph by Omar Al- tere are two allopatric species Iraq with the populations of N. wim^^motus t)emg*restricted mainly to the eastern and northeastern mountains of Sulaymaniya Province with notable o^centrations of populations in the Halabja and Pshdar dismcts. Halabja and Pshdar had six new loca- tions of the N. microspilotus with high number of eggs and larvae. The Penjwin district also appears to pro- vide many habitats for N. microspilotus. Schneider and Schneider (2011) reported N. microspilotus from three different localities in Penjwin, however, we believe that further surveys in Pshdar and Penjwin districts will re- veal new localities for N. microspilotus. The protection of known localities in these three districts is urgently needed to conserve N. microspilotus in Iraq. Populations of N. crocatus in Iraq are mainly re- stricted to the mountains and elevated grounds of Erbil and Duhok provinces, close to Iraq-Turkey border. Scat- tered populations of N. crocatus were located during our 2012-2013 surveys. However, many areas in the Erbil and Duhok provinces suspected to host N. crocatus have not yet been surveyed. Conclusion The mountain dwelling Neurergus newts are living in re- lictual aquatic environments which may make them par- ticularly vulnerable to environmental changes. However the geographical range and distribution of Neurergus newts in Iraq are not yet fully accessed and little is known about their ecology and conservation biology. From our survey results and a literature search we consider that habitat destruction including pollution when combined with climate extremes, and especially droughts, are the main threats to these newts. Urban expansion and rapid development combined with severe drought especially amphibian-reptile-conservation.org 046 July 201 3 | Volume 6 | Number 4 | e68 Al-Sheikhly et al. during the current years have impacted many fresh water springs, streams, ponds, and waterfalls which are consid- ered the main habitats for Neurergus newts in northern Iraq. Solid wastes produced by tourism and agro-chemi- cal pollutants, mainly from the use of agricultural pesti- cides and herbicides, are considered as the main pollut- ants that may impact Neurergus populations. Therefore, serious conservation actions should be ur- gently undertaken in light of various factors negatively impacting populations of these unique salamanders. The Iraqi government is responsible for protecting mountain biota including Neurergus spp., and particularly the Crit- ically Endangered N microspilotus, as they are iconic species for conservation. Greater international coopera- tion between researchers and conservation agencies in Iraq, Iran, and Turkey, countries sharing similar moun- tainous habitats and water resources, should be strength- ened in order to conserve the Neurergus species. The populations of N microspilotus and N crocatus and their unique habitat in the mountains of northern Iraq need to be urgently included in long-term monitoring programs with the aim of: 1) estimating the effective size and con- servation significance of genetically distinct populations; 2) quantify the main threats and gathering additional in- formation of the threats to salamander populations; 3) undertake in situ actions such as land/water mana; and protection; and 4) raising educational awa: should be prioritized to protect and conserve the g Neurergus in Iraq. Acknowledgments. — We than of Environment, Land and of Environment (IMoE) ture Iraq for their suppo Iraq (Kurdistan region Barbanera; Dr. MijAtar K Dr. Jorg Feryahqijrand Mr. Ali Ne assistance d Taha (Unive l, and Na- surve^l&Mi northern extend to^^. Filippo Elnaz N aj afimaj d, alaman for their Mrs. Zainfi Mahmod and Ali r their notes and photo- wa - Sarsank in Duhok for his helpful support to ran Nilson for his advice and ogical studies in Iraq. Literature Cited Al-Adhami MA, Hameed AK. 1988. Ultrastructural study of the thyroid gland of the salamander, Neurer- gus crocatus crocatus (Cope, 1862). Bulletin of the Iraq Natural History Museum 8: 25 4 1 . Allouse BG. 1955. A bibliography on the vertebrate fauna of Iraq and neighboring countries, reptiles and amphibians. Iraqi Natural History Mm mum Publica- tion 6: 1-23. Cope ED. 1862. Notes upon som^EFTILES o^ k Old World. Proceedings of the Accn^ry of Nature mSci- : 337-; on the . 96 pp. on SA. 1992. and Reptiles. and Reptiles, A systematic list of the verte- ral History Museum Publica- ences of Philadelphia KhalafKT. 1959. Reptile amphibians. Ar-Rc Leviton AE, Andemof^ Handbook to^mddle Society Study ol Oxforc MahdiN, ‘Iraq. 26: 1-104. ^The newt Neurergus crocatus (Cope, mqi Natural History Museum Publica- iaya U. 2010. A newly found locality of critically endangered Yellow Spotted Newt Neu- microspilotus (Nesterov, 1916), nourishes hope for its conservation (Salamandridae: Caudata). Zool- ogy in the Middle East 51:5 1-56. Nesterov PV. 1917 (1916). Tri novych chvostatych am- fibii is Kurdistana [Three new forms of amphibians from Kurdistan], Ezhegod. Museum of Zoology, Nauk Petrograd 21: 1-30. [In Russian]. Schneider C, Schneider W. 20 1 1 . The Kurdistan newts of the Genus Neurergus in Iraq (Caudata: Salamandri- dae). Herpetozoa 23(3/4): 3-20. Received: 14 May 2013 Accepted: 16 June 2013 Published: 15 July 2013 Omar F. Al-Sheikhly is an assistant teacher and wildlife expert at the Department of Biology, College of Science at the University of Baghdad. He has worked as a field team leader, wildlife expert, and provisional photographer in the Canada-Iraq Marshland Initiative (CIMI) and Nature Iraq. Omar has trained many Iraqi biologists who now work at the Iraqi environmental institutes such as the Iraqi Ministry of Higher Education (IMHE) and the Iraqi Ministry of Environment (IMoE) on wildlife field monitoring methodologies. He has studied and photographed most of Iraq wildlife rarities including Neurergus microspilotus. amphibian-reptile-conservation.org 047 July 201 3 | Volume 6 | Number 4 | e68 New localities of the Kurdistan and Lake Urmia newts Iyad Nader is a senior professor in mammals and wildlife expert worked in Nature Iraq Wild Mammals Advisory Team (NIMAT). His special interest is the conservation of the wildlife of Iraq and he has published numerous papers and conducted field research with the wild biota of Iraq. Nasrullah Rastegar-Pouyani earned his B.S. in zoology from Razi University Kermans hah, I ran, in 1986 and his M.S. in zoology from Tehran University, Tehran, Iran, in 1991, where he studied the agamids as the central object. He started his Ph.D. in Gothenburg University, Swedei^^rl994 uncAfche ad- visement of Professor Goran Nilson and graduated in 1999, working on taxonomy an^Kjgeography ol Plateau agamids, with Trapelus as the main objective. He is a professor and head of tl^fckdogy departf Razi University. His research interests include taxonomy and biogeography IraniaH^teau, the^ East, and central Asian herpetofauna. Robert Browne, director of the journal Amphibian & Rep A and practical experience in many research fields suppo sustainability. range of academic ion and environmental amphibian-reptile-conservation.org 048 July 201 3 | Volume 6 | Number 4 | e68 CONTENTS Administration, journal information (Instructions to Authors), and copyright notice Inside front cover Mozafar Sharifi, Hossein Farasat, and Somaye Vaissi — Sexual size dimorphism in Neurergus kaiseri (Cauda- ta: Salamandridae) in south-western Zagros Mountains, Iran 1 Serge Bogaerts, Henry Janssen, Jennifer Macke, Gunter Schultschik, Kristina Ernst, Francois Mail- let, Christoph Bork, Frank Pasmans, and Patrick Wisniewski — Conservation biology, husbandry, and captive breeding of the endemic Anatolia newt, Neurergus strauchii Steindachner (1887) (Amphibia: Cauda- ta: Salamandridae) 9 Nasrullah Rastegar-Pouyani, Mohsen Takesh, Akbar Fattahi, Marzieh Sadeghi, Fatemeh Khorshidi, and Robert Browne — Ecology of Kurdistan newt {Neurergus microspilotus : Salamandridae): Population and conservation with an appraisal of the potential impact of urbanization 30 Elnaz Najafi-Majd and Ugur Kaya — Rediscovery of the Lake Urmia newt, Neurergus crocatus Cope, 1862 (Cau- data: Salamandridae) in northwestern Iran after 150 years 36 Retracted: Omar Fadhil Al-Sheikhly, Iyad A. Nader, Nasrullah Rastegar-Pouyani, and Robert K. Browne — New localities of the Kurdistan newt Neurergus microspilotus and Lake Urmia newt Neurergus crocatus (Caudata: Salamandridae) in Iraq 42 Table of Contents Back cover VOLUME 6 2013 NUMBER 4