dhe
Hi
ati:
at ry
: ph be
iB}
statute 5
nie Mehta Toei Ht
t “
aye
bua bal
ui i sett Siete baatiat ee
: rt L cyeass gh te + it . tw
sii {nu clnib teh gt Srey bast
eiirers ry ere eeu stsrtnertts rt Mareret ieee astetres
Ua Bohne i i Mehta beset ae ats these bisa heg ;
Se ay rf thy Lyi ge : srt elgirer asesegs Peterson 5
ies ; ’ 1th peaks reheated Hot ee oteroene neta 7s ha by bys
shea ss beg tatpanai armani Eg tc
Becher a ne Hit sparking setae: tithe Raed Hy
cpp berertcy i Wirbhd, ; Pi tits ts garbaneectrrs tithe
intone 43 neve ae Rak ty eins inet
Peter Wee Lib abee reson 1; i arate rt att chy
EI Ryser ehate sen aI chee abet Hye Det th t Toe oe
sa Pathe ey a Rilateteeisee reer eee yay ys ot : Sih Saptyert te
yeast aeeg sot rst Bieta aie eaerE ore 2 ts print : erat st
tetera hh terabyte: Phebmertatrs eats! strrid gions: resi siapiggesyiety at rae
bet ties sedan rat tay Sts aris ety Bes ait Rite be sare eet + neuer is
te ey botnet serie ce t M43 0 oT a bre aye! 4 sigs, reits tery + i. i a ay
pbb teas arbors? - coleciarietigs or : 4h; ath ease Mibergh shot tthe
PEEEE PS Ey bitte 3 ae *y ite 1 Fst he ctahercye rte rstias rat
Septet tie f ents srrein RHitH iret hi tee gts re Samer rast
f yas are Pa TS attest! Lar tee. pevelite yest tate ceteed te ate te bts
nose gitsts attest ate atte : Lip tem ernest ttt icerttecaatsateer Sgt
yay a tipo igs 4 rhe: eter roietites eat reeseat te m : abbgtroys Stara atc agtescay arate Te nite
reat si ee eee bs : IMT Mor be ete bee aaNet ete ety Sey per titres orbs bebe ey ake rfehe re brs tebetesebe artes peer reeset ests
Cin sh eeyty eytyy re lehety pith fidbes ty atti + [afodeg nie Dt reyes eat " eat tet eet Tarte The es 8 he ogee ehhh ine pat
pais an peri epee ceca ae a pet autre tht SEE i
Sry , ; Soeape be oa he sees eee) i , Tyeseye ea t shat
por iiie Toes ere 4) ret tn rt aed peste
Miers ee) eet Piru ts ‘ yh at
Sata shy! yas hers Siphe wt ctr oF tubs teh yt % eyes martsatiro bh restates
maneentreeadhe tet i Haag by Petre i ry enuiis
Me eared Pedy wees et TO AILS ty , cots
paereerar est vay Peateby rune tester aoe oe reteea spurte weepcate teh taor
in. dave Se a See ee eye Sra a
SL aia cea ere ai cpbrbieeieghaet di ts ifs reubaie baatitbesrie sae itt are
Pee cr tin iteaye ¥ pebeaeeg Cate ia aoe Y epaiytiseetatemtee tee tetatyts tits
+ : ~ sat et ett cheat bese epee belatetct ie be ay
ha at tyt . y See eSetaeyt
pe ; 1S att med tntag bee tees
oH 4 ase ty Rduiietee be ieee eae
ws ryt Ste gehe pars Ane poo bee beet gee"
“4 M Te he ott oh rants pith tytts saebey
‘i ty Has ete, pH poate ea bit gtitenreesene bos!
oe bre Mest ahstete Dy Rugg steseveyan cesta pea ct
are LRA _— a reese aera redoetel Th Shas ba beng othe te et
Ser sci Ba Raitt i
uta bhatt
BE at oes Nd. i
Dene st it dior Fe Seg Wee be be tof 26409
sie OHHH i * a , ap eat Moe ht) ritet eid sea ie
4, ete de Lieder) re vite Garrett tera
; seat Sei sts Eater of ater by iereetets Berenihaatsieretee tts
bdichp she ld siete me Heiss oe + pate paitahs ty
i Osis YA eey stetyerteeet if rf ; arte ben bere the neay
: vce ‘ Li yest!
: i tear senda ah
ey; renee Meitic!
pita yt +A
ay tis ets Kei tiey
a et tae pieid tr, Sty
aria ay bereayleeytt hah i spat piteteettete!
wa Qe tit 7 ite pa betdoeit Rpts! pies os
~ thy } ry atehs obese Marthe ria sents tet ee be
fey AEE 3 es Way tr
ibys eater tete t
te
i is ios
t,o, Fp reer er
t
4 bse be
Lethe ye oe
bi ad be 5408 Seog Sheet Ste
Besesse Meret Petits hsaeae a
erste ‘ cu pey se
& byigetrhed tt yr i Lh oe
piers bs gh Thee a Pe eayt et
t z ayes 4d kegs;
3 bang Wand Mod 9
L Petpet
,
phe
r ° a
Baidvtetaty oe} he nae vy
ies ' pee
4 thy | pti
2 + a bee he
oy t
= 4
te or
2 7
Se 7 tatee oo eae
iene a ; Gat
fe ri steay| ttt
tes
Hidisiengeigy
Hh fsiate't}
HARVARD UNIVERSITY.
DIiBRA HY.
OF THE
MUSEUM OF COMPARATIVE ZOOLOGY
ASKS
GIFT OF
ange nA CN, ae
oul A a:
alte oT IM Ge
; ba
a Te Cla cn ihet ed
“i DL ies Te ak i
m4
. ;
ia - eo
Og
A LABORATORY MANUAL
INVERTEBRATE
ZOOLOGY
BY
GILMAN A. DREW, PH.D.
PROFESSOR OF BIOLOGY AT THE UNIVERSITY OF MAINE; IN CHARGE OF
ZOOLOGICAL INSTRUCTION AT THE MARINE BIOLOGICAL LABORATORY,
WOODS HOLL, MASSACHUSETTS
WITH THE AID OF
MEMBERS OF THE ZOOLOGICAL STAFF OF
INSTRUCTORS OF THE MARINE BIOLOGICAL
LABORATORY, WOODS HOLL, MASS.
PHILADELPHIA AND LONDON
W.B. SAUNDERS COMPANY
1907
a ” ae 4 a C.F
7 i y eye Rey “) Px
“2 ie. an Toner PISS ee loan .
Ne Ar ae Area,
“ ‘a - 7 ee i
¢ ets
«
‘
| CHA AL |
a: AENIDOS COG Cab
CAM SE0IKEN A:
Copyright, 1907, by W. B. Saunders Company
: F = : ‘
.
PRESS OF 54
W. G. SAUNDERS COMPANY ie -,4
PHILADELPHIA
&
— = ~ . ef he +
ae
& é a oem
= e ¥ a
PREPAGE,
THE present manual has for its basis a set of laboratory direc-
tions prepared by members of the staff of instructors to meet
the needs of the class in general zoology at the Marine Biolog-
ical Laboratory of Woods Holl, Massachusetts. Those who
were associated with me in the preparation of the first notes
were Dr. Robert W. Hall, Dr. James H. McGregor, Mr. Robert
A. Budington and Dr. Caswell Grave. Other members of the
staff who have either aided me in modifying the original notes
or who have added others are Dr. Winterton C. Curtis, Dr. D.
H. Tennant, Dr. Otto C. Glaser, Dr. Grant Smith, Dr. John H.
McClellan and Dr. Lorande L. Woodruff. Each year for the
past six years the directions have been changed where experi-
ences indicated changes should be made.
Probably few instructors will find it desirable for their stu-
dents to follow closely all that is given in this manual, but it has
seemed better to arrange the matter in a logical order, and in
some of the forms to call attention to only the important points
of anatomy or adaptation, than to try to make the directions
for each form complete in themselves. To make the directions
for each form complete would necessarily add much labor for
the student and would, by the repetition of well-known facts,
tend to blunt some of the new and important points to be gained.
The type method of laboratory study has for many years been
the prevailing method, but care needs to be exercised to keep
students from making everything conform to type, and in lead-
ing them to see the wonderful adaptations that fit the different
animals for their particular lives. The manual is not’ intended
to lead students to a knowledge of comparative anatomy alone,
but to an appreciation of adaptation as well.
It has fallen on me year by year to see that desirable changes
were made in the directions, and it has finally been my lot to
put them into their present form, but much of the credit be-
longs to the men who have been associated with me in the
instruction work at the Marine Biological Laboratory.
Tur AUTHOR.
May, 1907.
iii
CONTENTS.
PAGE
mR NS NS gio ass Bo ses. SS gece a hetend oa cee a enn WN 1
oT DO he Be aS hae hee a en a OY» LLY Wiehe SNE) 3
PUREE IOAMEINOUCULS 23 277 both unin e2.) 0 25.5 ee ae ae 3
SUCRE APIATTNRRE TH) ot Cony tn ee et eo kb ti Re ne Re ie 4
mevnospherium.or Actinopnrys .... 0: sence se ehone soe e- 5
SRR MIE eS PHTOEUAY 0 Ape Ete ecvtn tata rise hurt ant Mae Nm, ene ey oe 6
PREPS Gh ol Ned SA ewe Si rn ee ic tyes eh ee Oe SN et 6
MUA PsP ee Ra a cco BC. SA ae bli ee Oe OS d
“CT ETE TRIICLT EES Se gp Sit eS Re tel RRA SEE EAS BO lt C3 Tig Png 8
be ETT EVE te I iS ie a RCE ee eke tL aoe 2d 8
ERE PERMONT RE ORES erie scm a ee PR he fay eeu Me 8)
ere IMI Me EN eyo ts eh, ets rs Batia e ks Va a aes aaa
RESET NG ee tT re ary CPS SS cybhe Gy mee RS ty SN PLL. 10
SPOS ELITE) SPiN ee ae Pt EOD hd Ba caer AML Dt 10
Seema APRBEEA Seat SE Fcc, wee tee eee glee oe oe 11
na Pale teste cae Sete cee ie © eye a Pg uke Me rahe a) ) ny eR ae fat
OE TELE G8 15 a a a ce ea eee aang a PPR || 13
ED Cer Mitre Sais na sts ole ey oo ee 14
RerremA MEE tay et ar. eng il See ngae th eg ees Is ae Roe 15
MTR 8 x he ny lanl Be ean uy Phir. ay Rees oe ee 18
RR TMCIE NORA PhP Re ae Ie Nk em og nl vk Md wa 6k lee OO 20
Pence resn- water Pole py. oi. su cee yan ete 20
VESTER LS ee SEI ae OSG ea ge NT er Wal CR a 8) 2 22
ETE ELE DS ts Eek Re a BR oe cen RTT SOR TS Pa 24
Seana TTESERAL eer pcs oot Ee ia bt ks od aaa 25
STOTT 0 2 || ee ag Pee een es) TS MT ea aL 27
TITEL ESEECY CLES iy ISS geese ae ee ire Soka eeaMaie o=2e0 ONE a 27
UMEDA te acti aca ic ey Root nek, Nghe i 7S in awd Ca aT
PAN ee Llyn Bib oho aeee ns cea ita 5" eee eae 27
os SEES oR SR lg oa ne i eer el PS ae EN ee 30
PACT tha | SCR AMPIIONE) oo: 3.-s oS. 5 coh 5 i, eas 30
AME MTUVEL A Shc eet ra Ne cee ee Se ON pies) e 32
MIPS TAMMIsn etA Bae Se MES OAM OUND SoS oe OY 32
eM UTE NC EDEN 6 Sc. coi occ oes Res oe a he Sie oT Pek 35
SURE TEMRCI NETO E IES ily oy 22 ory ee ec hee ee iar Ls 36
rem E RU PRICLOCTULALAS ooh So hd LS ks RE eg ek NG Ln 36
InGcionra or Sy nccelidiim Fs. ek 2k athe ae Ss caea es s EY
Lo STE SLC WPT SONS of SIRE al crcl ee Se seit aad Ruane get inhale CNC Se 39
Enema olcecids (Piston) .ec.65 ss icles ox p aeenishee’s cists oe 2c 39
SPORES eet ot, iach ats re eS ee ee bey ee 41
Proccemoniriiii: ACMIAbUOE b, > <.o55 36-6 anv ea wre cleek aw erce hak 41
vi CONTENTS.
PLATYHELMINTHES (Continued). PAGE.
Wee TIA ta 2 oo hee bie hed ss Se ae 44
Petenstenwaa . os. fo a ce Seo we oe - sade 2 2 ee 44
NE MATHELMIN THES.) occ. 025 toes tea i 46
ane ARRE ores ae iv Soke Ss Lee Bea OE eas Be eS Oe 46
rel sa Se, Oe lee Sn y's Wa ees 47
TROCHMULMUN PRES... so fois bees os tot eb. he 5 os. he 48
Happen 2 oo eee ek See OI aa 48
Brachionus (A. Rotiier):.. 2 svelv22t. 222 =. >... Se 48
BTA AULD A. ee ee ee wate IRS 1 Sk 5 eo bole os, 50
POU cls) oh 4. odo Pe Ree ne eas Sod orn 50
Bra, 32 6. Gea eas tee ew awe oe a 50
Phimatella 3 Soh 8 ho es oo 52
RENEIOPODE:. 6. 544006 (0s a! sa ee a eee En 52
Tereiratialina. «2.00 ace 45 < oe so os oe eae nee ee 52
BOHINOMERMATA « boc: sus oo. Re ES OR ee 54
ASTERGIDEA™ oo s:c 0. oS oaks Soe hoe een 4 os ee 55
Asterias (Starlish) 220. ix. <2. ae neda- 2: oe 55
OPHTIBOIDEM. (40522. oe 2 dS ee oe eo 60
Ophiura (Serpent-Star) .-..-... 0... +. +++ +. +> + ee 60
FUCTINGIDEA ©. 0a5 20 oo = Kr debs oe one Gece ee er 61
Arbacia (Sea-Urehin) ...... 5... 5.2.0. os: - ne 61
FIGLOTEURGIDEA......0. 25526 02 ns bea Da on ee 67
Thyone (Sea-Cucumber)::....-.4.- ¢-.55. i. 6: 40 2 67
PU TOTON VIDA: ee he foe oe dw baleen oe ee ae ne ae ee 70
CraeTorObA.. =). 600 lois soho amas oe Le 2 71
Nereis virens (Clam-Worm)>...¢. 2... <... 1: 5. 4.342, 71
Lumbricus (Karthworm) .....:... 22... 202.2 ee 74
Autolytus cornutus 2...) 2... o. <2. 922+ ee ee 80
Lepidonotus squamatus ... 2... .. 2. 2.2.26) 33s 81
Diopatra cupreas... 25/2. .<%. 22 Jae 2% El eee 81
Chetopterus .. 25s 62 2 oe a OS le 82
Amphitrite ornata:. 0...) 0s02-25 V2 oie ee 83
Cistenides gouldit <2... 2. 22. de ee 84
Clymenella-torquata s- 2.2). as ae ee aes 84
Arenicola cristata... 2.2.5 62 4. ease sa 3s eee 85
Sabella microthalima ........... 5. 72 «+ sale ee 85
Hydroidés. . 222 ews on s'. See 3 Se ae ee 86
Spirorbis borealis ...... . 2.2.0... 3/25. Se 86
GEPHVREA . 2)... J. ee oc Pe eee See 86
Phascolosoma... 2.36. vcs 5 seis 62k 2 Oe ae ee 86
MOLITUSCA |... ce bd as 22 ee Oe 89
LAMELLIBRANCHIATA .. 2.60 oo ob nee Ae ee 91
Venus mercenaria (Quohog) ...... -... -.¢)55.+--- >> +- = ee 91
¥ oldia limatula... 3...) 542T 2. Uae 99
Mytilus or Modiola (Mussels)..2>. oc. ..0) 2: 37 >. 100
Pecten itradians (Scallop) ....2... 2. 22S 3.-8- 2s See 101
Ostrea virginiana (Oyster). 20.6... 4 eae 103
Dolenomysa «oo... ws se Se a ee ok 2 ee 1038
Mya arenaria (Long Clam) ... 2: i..03.< 27 2 se 104
Ensis direetus (Razor-shell Clam) .:......... 22s. sag)... spe
CONTENTS. Vil
Mouuusca (Continued). Pace.
oe DRE ie, Sane eats ek SN arte ee a RO 106
SU MRMEONIRCET eM Geete ox cy Ce EY har tg Ae xs rile 106
SLE EA tite Soa Sih Gh OR alr SR hei So RS es en 107
Pema ye Liane es oY LS. x See ge Aceh ab e's wht cine bn 107
MNP MarR GLENEEN) 52 Goo sock nee Merscere Ha ccie «se oe mle Dae 115
EN Fact ones 5 Ph ole Fk as oA De tlw sie adn 4 ae Ao 124
A SRMRINEE Sa Cpe ee Sn ES lath ores Gigs wea tee a ive SAA, Oe ee ee 128
onmiares amenecanns.(LObSter).. <--> «0... «ea dkms paeeedee
ialmecves nastatus¢(iiie Crab) i... .0.02<. 000 oo ee ee ee ee
Papacuras (liermn Gra)... 226 es ooo cece see OO ee ee 138
MRE PENG Sane occ 52 2's 15. oko s nice eaten te de Spe 139
ETE | Te Ry Ba ERATE a i pA RO Perr at en er gre LRT OTE AE 139
LS SCE RO ier tO Rat On ee Mee OR ead eT Lorine ent sor 141
alorenestia (eachah lea rst ace) Gala soo sa eee 141
Porceiho or Oniseus, (Sow-Bug) . 5... 00.7. e252 2c eee BAS
(WET DEE] [Pan eat ii aie ae an eee ool mananre i Same onece eS Waa ate ge 143
Drancaipus (lary Sirimip)s:. os.) 4). okies. sien oe
EC HIAS nega Son piso Naa te Oey SU Re ee eee ee 144
LOGUP TSS eee 2 or ea a oar ge ce Pema ae 145
Peereerae PENI MOISE a. oe og 5 oe 2 ak nite ae oe ee oa 146
Mepasat eee Pear NACle) 2502-25 cists sc heen ewe seake ees 146
PMR ETUAERSR vente, fry Oh nate re: eee gs HU See eae Pe et 147
Marmsiine (larmesiioe Cra). 2. .en Fs as oie se ees He eds ane 147
Pee ARERN SIRE FIRED oh 2 ccs teres eon sa Craie ey eee, DR 149
ipeira Cr OUHE- WER Opler) 22. toes ca Ss ome Pee SE ea eee 150
EDU TUES ER SO gene Pa eg eo a ae 152
MYRIAPODA.. Sty EON rca ee Meme Say ts 2
Lithobius (Centipede, Earwig). . IE SE Sn OE Re eH ge Ne 152
RE ET OUSARE IES i aes oo )-s ae ue wane eee cee ae oo eee 153
OR en ica as ge Bit hai ote en eae nat coe wo Sg eke 154
Peerig Hii (tTASBiGpPPe’) s.408 "San ain sca ee ae cs oe os 154
Peas melitieds (HoneyBee). oo: oa. ss" age nc oe ai ee 159
Pe ORE ee oe Shs Perce ee ig, Ue oats wn ged Baha PE eel ae DS 164
Demet Mey a ets ec saa x Sk ey yee RR ens od a ee 165
ives mame UEMOIS fc 6 nt ck Sa bh oe SSS Oe 165
REPRE Ss otic act re as crag ime ee Sagat: 2 Soy one cee 168
Rab nat cho Sgr ch Pe caer 1 Pee eon inl < SR EE 169
PEMIATOSCUIM (CAE OPK). 2.5 x sot Stas es gees 2s Oa tee ee 170
STUNDE, COOPAN LONI rok ta 5 2 acc 1 2 -0is Stags cc She oy Mies + Peotone somes 172
RURER MRT AR A 2 8 enn Wate et Be 8 abe NS Cone aa di No Ie ARE 173
te Wie) ol Si tasg) tol 28 ite ea ffs Bier gies eg mo REC ene ac ee oe SLB 173
a FOR GUIDANCE IN MAKING PERMANENT PREPARA-
$e REIS Se ar ig OA NR ne Ae Seer ST oe er ROR ar eo Sag aad ER 175
DEEN gto. co On ahs ok Soe aed ee ea eee eee ar a id aio rw oad 181
MNOS Se Se Sane whinge akg he a eR wae es s hehtate Sond = 195
INVERTEBRATE ZOOLOGY.
PROTOZOA.
Unicellular Animals.
Cuass 1. Sarcodina.
With changeable pseudopodia, during adult life.
Reproduction by simple division and by spore-
formation.
Subclass 1. Rhizopoda.
With lobose or reticulate pseudopodia.
Order 1. Amoebida.
With lobose pseudopodia. (Amoeba, Arcella,
Difflugia. )
Order 2. Reticulariida.
With fine branching and anastomosing pseudopo-
dia. Shells, when present, usually calcareous.
(Microgromia, the Foraminifera.)
Subclass 2. Heliozoa.
Typically spherical in form. The pseudopodia,
which radiate from the entire surface of the body,
are ray-like, seldom changeable, and usually pos-
sess an axial filament. (Actinophrys, Actino-
spherium, Clathrulina.)
hile 3. Radiolaria.
With ray-like pseudopodia, and with a chitinous
capsule inclosing the nuclei. The skeleton, when
present, is formed of silica or acanthin. All are
marine. (Thallassicolla.)
Cuass 2. Mastigophora.
Motile organs in the form of flagella. Repro-
duction by longitudinal division. Colony forma-
tion is frequent.
Subclass 1. Flagellidia.
With a definite anterior end on which there are
1 1
2 PROTOZOA.
one or more flagella. The members of one order
(Choanoflagellidia) have one or more collar-like
processes about the base of the single flagellum.
(Mastigamceba, Proterospongia, Euglena, Pera-
nema. )
Subclass 2. Dinoflagellidia.
Usually with two flagella, one encircling and
the other directed away from the body. (Peri-
dinium, Ceratium.)
Subclass 3. Cystoflagellidia.
With two flagella, one of which is modified into
a “tentacle,” while the other is short and con-
tained within the gullet. (Noctiluca.)
Ciass 3. Sporozoa.
Without flagella or cilia in the adult period of
the life-cycle. Reproduction is by spore-forma-
tion. All are endoparasites.
Subclass 1. Telosporidia.
Reproductive phase of the life-cycle is distinct
from, and follows the trophic phase.
Order 1. Gregarinida.
The young stages are intracellular parasites, while
the adults are free and motile in the digestive
tract or body-cavity of the host. Sporulation
occurs within a cyst during the free period of the
life-cycle. (Gregarina.)
Order 2. Coccidiida.
Without a free and motile adult stage. Sporula-
tion occurs within a cyst, during the intracellular
period of the life-cycle. (Coccidium.)
Order 3. Hzemosporidiida.
Living chiefly in the blood-corpuscles of verte-
brates. In many forms the entire sexual
period of the life-cycle takes place in an in-
termediate host, as the mosquito. (Laverania
malariz.)
Subclass 2. Neosporidia.
Reproduction takes place during the trophic
phase of the life-cycle.
Order 1. Myxosporidiida.
The initial free stage occurs in the tissues or the
cavities of the organs of the host. The adult form
is amceboid. (Myxidium.)
AMCBA PROTEUS. 3
Order 2. Sarcosporidiida.
The initial stage of the life-cycle occurs in the
muscle-cells of vertebrates. (Sarcocystis.)
Cuass 4. Infusoria.
With motile organs in the form of cilia during all
or part of the life-cycle. Nucleus dimorphic
(macronucleus and micronucleus). Reproduction
is by simple transverse division or by budding.
Subclass. 1 Ciliata.
With cilia throughout the life-cycle.
Order 1. Holotrichida.
The cilia are of approximately equal length and
thickness and equally distributed over the body.
Trichocysts are present. (Prorodon, Parame-
cium.)
Order 2. Heterotrichida.
With a uniform covering of cilia, together with
an “‘adoral zone” formed of cilia fused into mem-
branelles. (Spirostomum, Stentor, Halteria.)
Order 3. Hypotrichida.
The cilia are limited to the ventral surface of a
dorso-ventrally flattened body. Cilia often fused
into cirri, membranelles, ete. (Oxytricha, Pleuro-
tricha, Kuplotes, Peritromus.)
Order 4. Peritrichida.
More of less bell-shaped in form. Cilia usually
reduced to those constituting the adoral zone.
(Vorticella, Zoothamnium, Lichnophora.)
Subclass 2. Suctoria.
Usually possessing cilia only during the embry-
onic stages of the life-cycle. Tentacles adapted
for piercing and sucking are present. (Podo-
phrya, Ephelota, Acineta.)
SARCODINA.
AMOEBA PROTEUS.
Amcoebe are usually easily discernible under the low power of
the microscope as irregular, semi-transparent, granular bodies.
Find a specimen in the material provided, which is known to con-
tain amcebe, and determine the following points:
1. With the high power observe the peculiar method of loco-
4 PROTOZOA.
motion, the constant but slow change in the shape of the body
by means of projections, pseudopodia, or “false feet.”
Make sketches at intervals of one or two minutes to show the
changes in the form of the body.
2. Observe the peripheral zone of hyaline protoplasm, the
. ectoplasm, and compare this with the inner protoplasm, the endo-
plasm. Observe in detail the formation of a pseudopodium.
Does the endoplasm extend into the pseudopodium? Can you
explain how the movement is caused?
3. Find a clear space which appears and disappears at inter-
vals; this is the contractile vacuole. Determine the length of
time between successive contractions. Are the intervals regu-
lar? When the vacuole contracts what becomes of the contents?
Do you know its supposed function?
4. Note the oval or rounded nucleus moving with ha flowing
endoplasm. What is its structure?
5. Food materials in process of digestion are readily seen.
Of what do they consist? They are contained in gastric vacu-
oles. By careful watching, it is often possible to observe the man-
ner in which food is ingested and the manner in which the undi-
gested matter is egested.
Make a careful drawing of an Ameba.
Ameoebe of various kinds represent in many respects the
simplest type of protozoan, and are therefore placed in the first
class of these animals, the Sarcodina. The individuals of this
class all possess pseudopodia, but many are quite unlike those of
Amoeba. Look over the figures of various Rhizopoda.
If time and material permit, study Ameba verrucosa, Arcella,
and Difflugia, and compare them with Amceba proteus. Do you
understand how the shells of the last two genera are made, and
of what service they are? Why are not shells good for all forms?
Drawings of these forms are desirable.
FORAMINIFERA.
With very few exceptions Foraminifera are marine and pro-
vided with shells. Empty shells from deep-sea dredgings or from
ACTINOSPH/RIUM OR ACTINOPHRYS. 5
the sand beaches of such islands as the Bermudas may be had
for study. Examine them with a low power by reflected light.
1. Carefully examine various shells, compare them with each
other and with figures. Notice the great variety in size and
shape and determine how the chambers must have been added
during growth.
2. Observe a single opening in a shell, and determine whether
the general surface has any finer perforations. Be sure to under-
stand the relation of the live animal to the shell. (Refer to
Calkins, pp. 71-78, for a general discussion of the shells of the
Sarcodina. )
Make drawings of several types of shells.
ACTINOSPHAERIUM OR ACTINOPHRYS.
Find, as usual, with the low power, and increase the magni-
fication as occasion demands.
1. Note the many fine radiating pseudopodia. These are quite
stiff compared with those of Ameba and for a considerable time
show little change, not being pushed out and retracted constantly
asin Ameba. Is the animal flat or spherical?
2. Both ectoplasm and endoplasm are so filled with vacuoles
that they present a frothy appearance characteristic of most
Heliozoa. The endoplasm of all Protozoa is alveolar in struc-
ture, but in Actinospherium the vacuoles are exceptionally
large, though not as large as those in the ectoplasm.
3. The nucleus is present in the center of the organism, but
it is somewhat difficult to demonstrate in the live animal.
4. At some point near the periphery, the contractile vacuole
can usually be seen. When it is found notice its action, and
immediately after it has contracted look among the pseudopodia
of that region for indications of its extruded contents.
Draw a specimen, indicating all of the points observed.
5. When the contractile vacuole discharges, or when any
foreign body touches the ends of the pseudopodia, notice the
way in which this type of pseudopodium is moved. What does
this indicate in regard to its structure? How far do the pseudo-
6 PROTOZOA.
podia extend? They may be seen to contain minute granules
when studied with the high power and best hght.
6. If possible, observe the process of catching food with the
tips of the pseudopodia and the manner in which it is drawn
toward the body. Note any motion on the surface of the body
as the food is drawn closer, and also the manner in which the
food is finally ingested. Are there any indications that the
pseudopodia extend as still finer filaments beyond the point to
which it is possible to trace them with the highest magnifica-
tion at hand? If the capturing of food is observed, make a
series of diagrams to illustrate the process.
If possible, observe a specimen undergoing division. Draw.
It is desirable to examine Clathrulina, noting the stalk and
skeleton. Look over figures.
MASTIGOPHORA.
EUGLENA,
Understand its habitat and with what forms it is usually
associated.
1. Observe the free-swimming movements of the organism,
and the euglenoid changes in the form of the body. .
Make drawings showing the changes in the shape of a single in-
dividual.
2. Distinguish anterior and posterior ends. Is there any
dorso-ventral differentiation? Note the motile organ, the flagel-
lum. Where is it attached? What relation does it bear to the
gullet? How is it directed during locomotion of the organism.
Does it serve any other purpose besides locomotion?
3. The green color of Euglena is due to chlorophyl, and this
enables the animal to live in the clearest water, being nourished
like a typical green plant, but minute particles of food are also
taken into the endoplasm through the gullet, and thus Euglena
combines holozoic and holophytie methods of nutrition. Consider
the bearing of this on the position of Euglena and its allies in
the protozoan scale.
VOLVOX. 7
4. Note the absence of color near the anterior and posterior
ends of the organism. Near the anterior end also notice the red
pigment spot, or stigma. What is its probable function?
5. Stain a specimen with iodin and look for the nucleus.
It is obscured by the chlorophyl.
Make a drawing showing all of the points observed.
Look through the stock cultures for other forms of Masti-
gophora, such as T’rachelomonas, Peranema, Phacus, ete.
It is desirable to make drawings of the different forms.
VOLVOX.
Volvox globator is better for study than V. aurens. It may
be distinguished from the latter by the larger size of the colony,
the greater number of cells that compose it (about 15,000), the
angular shape of the individual cells, and the stout connecting
processes of protoplasm, into which chromatophores may enter.
Observe the movements of colonies in a watch-glass of water,
with the naked eye and with a low power of the microscope.
1. Do the colonies tend to collect toward a particular side
of the dish? What reason is there for the reaction?
2. Place a number of colonies on a slide with enough water
to allow them to be covered without crushing them. Study
first with the low and then with the high power and determine
the species. Understand the relation of the individual cells to
the colony. (See Parker and Haswell, Fig. 50.)
Draw a figure showing several cells and their protoplasmic con-
nections.
3. Compare in detail an individual cell with Huglena.
4. Observe, if possible, certain cells, called parthenogonidia,
which are specialized for asexual reproduction. These divide and
form the daughter colonies, which become detached and swim
in the interior of the parent colony. They are finally liberated
by the rupture of the wall of the parent colony.
Make a figure of a parent colony that incloses several daughter
colonies of different sizes.
8 PROTOZOA.
5. V. globator is moneecious. Look for eggs and bundles of
spermatozoa.
Figure them.
6. Be sure to recognize the significance of the fact that the
cells of Volvox are differentiated into somatic and germ cells,
and to understand the resulting physiological division of labor.
(See Calkins, p. 232.)
7. Consider the reasons for and against regarding Volvox
and allied organisms as animals rather than plants.
CERATIUM.
1. Examine this form with a high power, and in a favorable
specimen notice the sculptured outer surface of the cellulose
test. The living animals are green or brown owing to the pres-
ence of chromatophores in the protoplasm.
2. Note the furrow encircling the body. Does it extend
completely around it? Is there a short furrow on one side at
right angles to the first, or a depression of considerable size?
Understand the position of the flagella.
Draw the animal, showing the points observed.
Look for examples of the earlier stages of division, and of
later stages, which appear as chains of fully formed individuals
attached together.
NOCTILUCA.
If living specimens are not to be had for study, material
preserved in alcohol, after suitable fixation, can be used. Spec-
imens are best examined in a cell-slide under a cover-glass.
1. Observe the nearly globular shape, and on one side a groove
from which arises a large flagellum or “tentacle.” Is there a deep
groove near it?) At the bottom of this groove it is possible to see
the mouth in a living specimen. Another smaller flagellum is
visible in living specimens inserted at the bottom of the mouth,
but in preserving the organism it is usually destroyed.
2. Note the appearance of the preserved protoplasm. The
endoplasm appears parenchymatous. At one point a more com-
ee
GREGARINA. 9
pact mass is seen, from which strands appear to radiate. This
has been found to contain the nucleus.
Noctiluca is phosphorescent, and frequently causes very bril-
liant displays.
Make a drawing.
SPOROZOA.
GREGARINA.
Remove the head and posterior end of a larval or adult
meal beetle and pull out the digestive tract with a pair of for-
ceps. Place the digestive tract on a slide, split it open length-
wise with a sharp scalpel, and then spread it out, with the
inner wall exposed, and cover. The operation should be per-
formed rapidly to prevent the material from drying. If the
beetle is infected, numerous gregarines will be visible under the
microscope. Study with low and high powers.
1. Does the animal move? A great number of refractive
granules are present in the protoplasm. They are regarded as
reserve nourishment. ‘They can be removed with acid.
2. Note that the body is covered with a membrane, and is
divided into a dense superficial layer, the ectoplasm, and a cen-
tral, more fluid mass, the endoplasm.
3. The endoplasm is separated into two parts by a portion
of the ectoplasm. The anterior part is termed the protomerite,
and the posterior part the dewtomerite. In which is the nucleus
situated?
4. Is it possible to distinguish a layer of myonemes just ex-
ternal to the endoplasm?
5. Is there another section of the body just anterior to the
protomerite? If so, this is the epimerite.
Before reproduction Gregarina throws off the epimerite, leaves
it in the cell-host, and falls into the lumen of the digestive
tract. It then encysts, and the protomerite and the deutome-
rite form one spore-producing individual. The attached stage
in the life-history of Gregarina is termed the cephalont, and the
detached stage, the sporont. (See Calkins, Fig. 77.)
Make a drawing.
10 PROTOZOA.
INFUSORIA.
PARAMECIUM.
Place a drop of the culture on a slide, cover, and examine
with the low power.
1. In an animal not closely confined note the shape and
movements. Is it possible to distinguish an anterior and a
posterior end? A forward and backward movement? Is one
side of the animal kept constantly uppermost? Is there a dorsal
and ventral surface? Do the animals change their shape either
permanently or temporarily? Individuals tend to collect about
air-bubbles and at the edge of the cover-glass. Why?
Indicate by a sketch all the points which can be determined
with the low power.
2. Draw off all superfluous water by means of filter-paper,
add a trace of powdered carmine, and then find a specimen
which is narrowly confined and examine it with the high power.
The particles of carmine are taken into the body. Deter-
mine how and where. Note that the carmine collects in gastric
vacuoles. What do you think is probably the nature of the
fluid in the vacuoles? In watching them do you notice any
definite movement of the protoplasm? Try to see the undi-
gested material ejected.
3. Determine the arrangement of the cilia, and the nature
of their motion. Is there a reversal of the direction of the stroke,
etc. ?*
4. Observe the contractile vacuoles. How many are there?
Is their position constant? What is their action? In com-
pressed specimens the contractile vacuoles and their reservoirs
are usually conspicuous. Note the order of appearance and
disappearance of the vacuoles and reservoirs.
5. Focus carefully on the margin of the body and note a very
thin outer cuticle. A thick layer, the ectoplasm, devoid of gran-
‘It is possible to decrease the rate of movement of both animal and
cilia by placing it in a solution of gum arabic. Specimens so treated
remain alive for some time.
—— eer ee
SPIROSTOMUM. VORTICELLA. 11
ules but containing radially arranged, minute, oval bodies, the
trichocysts, is just internal to the cuticle. The inner mass of
protoplasm, containing the contractile and gastric vacuoles,
and small granules, is the endoplasm.
6. If possible distinguish the clear, centrally located nucleus
(macronucleus).
Make a sketch showing all oj the above points.
7. Kill the animal by running a drop of methyl-green under the
cover-glass. What happens to the cilia? To the trichocysts?
Sketch the trichocysts with the threads protruded, and also note
and sketch the macronucleus and the micronucleus.
8. Observe, if possible, animals dividing and conjugating.
9. Study demonstrations of permanently stained specimens
for finer structure.
SPIROSTOMUM.
1. Compare Spirostomum with Paramecium, noting the
method of locomotion, the shape of the body, the ciliation, the
buccal groove and mouth, and the large excretory reservoir, fill-
ing the posterior end of the body and in communication with
the anterior end of the body by a canal.
2. Note the highly refractive, long, band-like (monilijorm)
macronucleus. In another species of Spirostomum the macro-
nucleus is similar to that of Paramecium. It is desirable to
examine stained specimens of the two species of Spirostomum.
3. Note the sudden contractions of the body. When these
occur spiral lines appear on the surface. Can you distinguish
these lines when the animal is extended? These are primitive
structures (myonemes) functioning as muscles.
Make a drawing of the extended animal and a diagram shou-
ing the form when contracted.
VORTICELLA.
Place a number of individuals on a slide and cover loosely
to avoid crushing. As usual, study first with the low power and
then with the high.
1. Notice that the body of Vorticella has the general shape
12 PROTOZOA.
of an inverted bell. The covering of the body is a very thin
transparent layer, the cuticle, underneath which is the periphe-
ral layer of ectoplasm enveloping the more fluid and granular
endoplasm.
2. The peristome is the rounded rim about the base of the bell.
3. The elevated and inclined area included within the peri-
stome, and ciliated around the edge, is the disk. It is some-
what convex.
4. The marked depression between the disk and the peri-
stome is the vestibule. It is also lined with cilia. The vestibule
defines the ventral surface of the animal.
5. The guilet, a slender canal, leads from the vestibule toward
the center of the body.
6. The anus occurs at the side of the vestibule. It is a tem-
‘porary opening from which the undigested products are passed
into the vestibule and so to the exterior.
7. Within the endoplasm are situated the clear contractile
vacuole, several gastric vacuoles, the long U-shaped macronucleus,
and the small round micronucleus. The macronucleus may be
made more distinct by treating with methyl-green.
8. The stalk is composed of a sheath, which is continuous with
the cuticle of the body, and, within the sheath, the contractile
axis or myoneme, which is continuous with the body ectoplasm.
Notice that this myoneme is situated within the sheath in a
very loose spiral, and that the stalk quickly contracts into a
close spiral when the animal is stimulated. Observe also the
manner in which the peristome folds over simultaneously with
the contraction of the stalk. What purpose does the contrac-
tion of the stalk serve?
Vorticella is distinguished from its allied genera by its sim-
ple unbranched stalk and also by the spiral form assumed by the
contracted stalk. In which order of the Ciliata does the cilia-
tion of Vorticella place it?
Make a drawing of an expanded individual and a sketch to
show the condition when contracted.
9. Study, by means of finely powdered carmine, the vortex
OXYTRICHA. 13
currents set up by the cilia. Note how the particles are collected
in the gullet, and at intervals are forced in rounded masses into
the endoplasm to form gastric vacuoles. Is there a definite
circulation in the endoplasm?
10. Endeavor to find several stages of reproduction by divi-
sion.
Large fresh-water species of Vorticella are preferable for
study, but marine species may be substituted when necessary.
If time and material permit, study Lichnophora, a marine peri-
trichous form parasitic on Crepidula. (See Calkins, p. 208.)
OXYTRICHA,
Infusoria belonging to the genus Oxytricha, or the genera
Stylonychia, Pleurotricha, Euplotes, etc., may be used for the
following study. ‘These forms belong to the order Hypotrichida.
Hypotrichous forms are the most highly organized of the class
Infusoria, as well as of the entire phylum of Protozoa, and pre-
sent a complexity of structure and function which is not found
probably within the limits of a single cell elsewhere in the animal
series.
1. In an animal which is becoming quiet, note the mode of
locomotion, the shape of the body, the buccal groove, the con-
tractile vacuole, etc., as in other forms studied. Compare the
ciliation with that of other forms. Refer to Calkins, Fig. 98,
and understand the relation of cirri, membranelles, ete., to cilia.
Draw, showing the structure in detail.
2. Run some methyl-green under the cover-glass. What
is the shape of the macronucleus? The shape varies considera-
bly in the different genera. Is it possible to distinguish the
macronucleus?
3. Prepare a fresh slide and observe in detail the character-
istic movements and manner of creeping over various objects.
As the animal turns sidewise, note the marked dorso-ventral
compression of the body.
Represent this diagrammatically beside the previous drawing.
It is desirable to examine permanently stained preparations
for division stages, finer details of the nuclei, etc.
PORIFERA.
Cells not differentiated to form definite organs. Water
admitted through surface pores and ejected through an osculum
or through oscula.
Ciass 1. Calcarea.
With a skeleton composed of calcareous spicules.
Subclass 1. Homoccela.
With the gastreal layer continuous so the col-
lar cells line the whole gastreal cavity. (Leu-
cosolenia. )
Subclass 2. Heteroccela.
Gastreal layer discontinuous. Collar cells restrict-
ed to the flagellated chambers. (Grantia.)
Cuass 2. Hexactinellida.
With a skeleton composed of siliceous six-rayed
spicules.
Order 1. Lyssacina.
Spicules separate or becoming united. (Euplec-
tella.)
Order 2. Dictyonina.
Spicules united from the first into a firm frame-
work. (EKurete.)
Ciass 3. Demospongie.
Great diversity of structure. Dominant forms
of today.
Subclass 1. Tetraxonida.
_ Typically with four-rayed spicules. (Corticella.)
Subclass 2. Monaxonida.
Simple, usually unbranched spicules. Spongin
frequently present. (Cliona, Suberites, Chalina,
Spongilla.)
Subclass 3. Keratosa.
Skeleton of spongin fibers. No true spicules.
(Euspongia, Aplysina.)
Subclass 4. Myxospongida.
Without skeleton. (Oscarella.)
14
GRANTIA. 15
GRANTIA.
This form is quite common along the New England coast,
where it occurs attached to rocks, seaweeds, and submerged
woodwork from just below the lowest tide-mark to a number of
fathoms in depth. You should visit an old wharf where speci-
mens may be found, and study their relation to the forms with
which they are associated. Specimens will be found to vary
considerably in size. The largest sometimes reach an inch in
length.
1. Examine a dry specimen and notice its general shape,
manner of attachment, and osculum. The osculum is surrounded
by a funnel of rather long spicules. Distributed over the gen-
eral surface, more or less hidden by the numerous spicules, are
many small pores. Their presence may be demonstrated more
satisfactorily later.
2. Look for indications of budding. If your specimen does
not show this, examine others.
Make an enlarged drawing of a sponge.
With a razor or sharp scalpel cut a dry specimen into halves,
with a stroke from base to osculum, and notice:
3. The central cavity or cloaca.
4. Many apopyles, the inner openings of tubes that are em-
bedded in the walls of the sponge, will be seen opening into
the cloaca, Are the apopyles arranged in any order?
5. With the low power of your microscope (with the light
turned off) examine the cut wall and find that it is traversed by
parallel tubes. Determine that these tubes are of two kinds.
(a) Regular, nearly cylindrical tubes that open into the
cloaca through the apopyles and that bear tufts of spicules on
their closed ends, at the surface of the body. These are the
radial canals. It is frequently hard to see their openings into
the cloaca, as the apopyles are narrow, so the section only occa-
sionally passes through them.
(b) Smaller and less regular tubes that open on the outer
surface between the clusters of spicules, and do not open into
16 PORIFERA.
the cloaca. These are the incurrent canals. In life there are
small pores, prosopyles, that open from the incurrent canals
into the radial canals. These openings are very minute and are
apparently capable of being closed. They are never visible
in dried material.
6. Examine thin, transverse sections of a dry sponge and
determine the positions of radial and incurrent canals.
Make a drawing that will show the arrangement of the canals.
7. Examine the spicules and determine their positions as
regards canals. Boil a portion of a sponge in caustic potash
until only the spicules remain and examine the spicules. See if
more than one kind occurs.
Draw specimens of the spicules.
LIVING AND SECTIONED MATERIAL.
1. Place a living sponge in a watch-glass of sea-water, add a
little powdered carmine, and examine it with the low power of
your microscope for currents of water. See if particles are mov-
ing in a definite direction near the general surface and near the
osculum.
2. With a moderately sharp razor cut tangential sections
of the wall, as thin as possible, mount in sea-water under a cover,
and examine with a low power. This will show both incurrent
and radial canals in cross-section. How can you distinguish
one from the other? In a favorable place look for moving
flagella. Are flagella in all of the canals? In favorable situa-
tions it can be easily seen that the cells that have flagella possess
collars also. (Collars may be withdrawn by cells so they pro-
trude but slightly.) You see now what causes the current of
water. Do you understand how a sponge feeds? Compare the.
choanocytes of the sponge with choanoflagellate protozoans.
Make a drawing showing the arrangement of choanocytes.
Examine transverse sections.of a specimen that has been
decalcified and stained.
1. The cloacal chamber is lined by a pavement of epithelium.
2. The radial canals are lined by more conspicuous cells,
the gastral epithelium, or choanocytes.
GRANTIA. 17
3. The incurrent canals and the outer surface of the sponge
are covered with flattened cells, the dermal epithelium.
4. In a part of the section where a considerable area of choan-
ocytes appear in surface view, look for the prosopyles, through
which the water passes from the incurrent to the radial canals.
(They may not be found.)
5. Make out any structures you can in the area lying between
the dermal and gastreal layers. What cells are found here?
Make a drawing of several adjacent canals to show the above
points and indicate the course of the water by arrows.
6. In the stained sections, look for single ova and for spheres
containing many spermatozoa, the sperm-spheres. Look also
for segmenting eggs, which are frequently to be found. The
ova are fertilized while still lying where they have developed,
just within the choanocyte layer. Remaining in place, they
undergo cleavage and develop so far as the amphiblastula stage
(see figures in the text-books). They then break through the
choanocyte layer into the radial canals and pass out with the
current of water. Living specimens are frequently found with
such embryos issuing from the osculum in the outgoing current
of water. The sperm-spheres, when fully developed, also break
through the choanocyte layer and, separating into their com-
ponent spermatozoa, pass out with the outgoing water.
Ova and sperm are present in the same individual, and the ani-
mal is therefore hermaphroditic. Whether self-fertilization is pre-
vented, as in many other hermaphroditic forms, by the ripening
of one element before the other, has not been fully established.
Ij the time allows, draw ova, sperm-spheres, segmenting eggs,
and embryos.
It is desirable to examine specimens of Lencosolenia, a still
simpler sponge, and of some of the more complicated forms,
like commercial sponges, Spongilla, Cliona, and Chalina. Why
is more than a single osculum desirable in such forms? Under-
stand the relation of the internal structure of the complicated
forms to the more simple forms. What reason is there for the
complication?
2
COELENTERATA.
With a single continuous ccelenteron or gastro-vascular cayv-
ity. With the exception of the Ctenophora all have nettle cells.
There are two cellular layers and a mesoglea.
Cuass 1. Hydrozoa.
Ccelenteron simple, without septa. Gonads usu-
ally ectodermal. Fully formed medusze have a
velum.
Order 1. Leptoline.
With a fixed zodphyte stage.
Suborder 1. Anthomedusz.
Without hydrothece or gonothece. The medusa
bears gonads on the manubrium. (Hydra, Pary-
pha.)
Suborder 2. Leptomedussze.
With hydrothece and gonothece. The medusa
bears gonads on the radial canal. (Obelia, Goni-
onemus. )
Order 2. Trachyline.
Without fixed zoophyte stage.
Suborder 1. Trachymeduse.
Tentacles from the margin of the umbrella.
Gonads on the radial canals. (Petasus.)
Suborder 2. Narcomeduse.
Tentacles from the exumbrella. Gonads on the
manubrium. (Atginopsis.)
Order 3. Hydrocorallina.
Massive calcareous exoskeleton. (Millepora.)
Order 4. Siphonophora.
Pelagic. Colonial. Colony usually shows extreme
polymorphism of its zodids. (Physalia.)
Ciass 2. Scyphozoa.
Body-wall of polyp thrown into four ridges (tzeni-
oles) which project into the ccelenteron. Medusz
without velum and with gastric tentacles.
Medusoid form predominating.
18
CCELENTERATA. 19
Order 1. Stauromedusz.
Conical or vase-shaped umbrella. No tentacu-
locysts. (Tessera.)
Order 2. Peromeduse.
Conical umbrella with transverse constriction.
Four inter-radial tentaculocysts. (Pericolpa.)
Order 3. Cubomedusz.
Four-sided umbrella. With per-radial tentaculo-
cysts. (Charybdea.)
Order 4. Discomeduse.
Saucer-shaped umbrella. Per-radial and inter-
radial tentaculocysts. (Aurelia.)
Cuass 3. Actinozoa.
With a stomodzeum, and with mesenteries ex-
tending into the ccelenteron. Fixed forms.
Subclass 1. Zoantharia.
Mesenteries and tentacles usually very numerous.
Order 1. Actiniaria.
Usually single. No skeleton. (Metridium. Sa-
gartia.)
Order 2. Madreporaria.
Usually form colonies and always have calcare-
ous exoskeleton. (Astrangia, Orbicella, Mean-
drina. )
Order 3. Antipatharia.
Tree-like. Mesenteries and tentacles compara-
tively few. Chitinoid skeleton. (Cirripathes.)
Subclass 2. Alcyonaria.
Mesenteries and tentacles eight in number. Ten-
tacles branched.
Order 1. Aleyonacea.
Skeleton in the form of small, irregular bodies,
frequently calcareous spicules. (Aleyonium,
Tubipora.)
Order 2. Gorgonacea.
Tree-like, with calcareous or horny exoskeleton.
No syphonoglyphes. (Gorgonia.)
Order 3. Pennatulacea.
Colony with one end usually embedded in the
sea-bottom. (Pennatula, Renilla.)
Ciass 4. Ctenophora.
Single. Pelagic. Eight rows of meridional
swimming plates. No nettle cells.
20 CCELENTERATA.
Order 1. Cydippida.
Nearly circular. Two tentacles, each of which
may be retracted into a sheath. (Pleurobra-
chia, Mnemiopsis.)
Order 2. Lobata.
Compressed in the vertical plane. Two large
oral lobes. No tentacle-sheaths. (Deiopea.)
Order 3. Cestida.
Ribbon-shaped. Two tentacles with sheaths, and
numerous other tentacles. (Cestus.)
Order 4. Beroida.
Laterally compressed. Without tentacles.
(Berce.)
HYDROZOA.
HYDRA. (Fresh-water Polyp.)
Hydra, the only common fresh-water ccelenterate, is fre-
quently found in jars of water taken from quiet pools or sluggish
streams that contain lily-pads, decaying leaves, and other vege-
table matter. The animals may frequently be found by examin-
ing the surfaces of submerged leaves, but it is usually better
to allow such material to stand in glass jars for a day or two,
as the animals then tend to collect on the lighter sides of the
vessels. They are easily kept in balanced aquaria.
Examine specimens in an aquarium and find what you can
about their mode of life. Do they form colonies?
Place a specimen in a watch-glass of water and examine it
with a lens.
1. What is its shape and color? Is it attached? If so, by
what part of the body? Notice the circlet of tentacles. How
many are there? Compare notes with others and see if all have
the same number. How are they placed?
2. Does the Hydra move its body or tentacles? Is it sensi-
tive? How do you know?
3. Examine with a low power of the microscope and review
the above points. You may also be able to see the mouth around
which the tentacles are arranged.
HYDRA. a
Make two drawings, one showing the animal expanded and
the other contracted.
Place your specimen on a slide under a cover-glass that is
supported by the edge of another cover-glass, so it can be exam-
ined with a high power. Be careful not to crush it. Notice:
4. The outer layer, ectoderm. What is its color? Is it
continuous over the whole outer surface? Does it vary in thick-
ness? Are the cells of which it is composed apparently all alike?
5. The inner layer, endoderm. What is its color? If color
is present, is it evenly diffused or is it collected in special bodies?
Are the cells of which the endoderm is composed apparently
all alike? Do they differ in appearances from those of the ecto-
derm other than in color? If the specimen is not deeply colored,
look for flagella moving in the internal cavity.
6. Examine the ectoderm of the tentacles carefully and
notice that each of the large, rounded, clear cells, the nematocysts,
shows a rather indefinite streak running from its outer end,
back into the interior. See if you can find the trigger (enidocil)
on any of these cells.
Draw a portion of a tentacle showing the distribution oj the
nematocysts.
7. Place your specimen under the low power of the micro-
scope, carefully run in a drop of saffranin, and see if any of the
nematocysts are discharged when the saffranin touches them.
Examine with a high power and notice the appearance of the
thread. Notice the change in the shape of the nematocysts
that have discharged. See if you can find two kinds.
Make an enlarged drawing of an exploded nematocyst.
8. Examine prepared transverse sections of Hydra. Notice
that the body is composed of two layers of cells, between which
is an almost structureless thin layer. Do the cells of the two
layers differ in size, shape, and structure? Do you find more
than one kind of cell in each or either of these layers? Where
are they? What are they?
Make a careful drawing of the section showing the arrangement
as you see it.
22 CCQELENTERATA.
Examine longitudinal sections, for differences in the char-
acter of the ectoderm and endoderm in different parts of the
body.
9. Reproduction. Examine living specimens in a watch-
glass of water for bud formation and for sexual organs. Sperm-
aries are just beneath the tentacles; ovaries, lower down;
buds may be found at different levels. What layers of cells is
involved in the formation of each of these?
Eggs are not formed at all seasons of the year and vary
greatly in appearance according to their stage of development.
Make drawings of the stages of reproduction that you find.
OBELIA.
These small, colonial animals are. common on submerged or
floating wood, stones, and seaweeds, where the water is rather free
from sediments. With the aid of a glass-bottomed pail they,
in company with many other forms, may usually be seen about
old wharfs.
Note the appearance of large colonies of this form that are
growing on stones or on pieces of board.
1. Notice the tree-like form of any single stem. Do the
branches have a definite size and arrangement?
2. At the extremities of the branches are the single individuals,
hydranths or zodids. Each is similar to a single Hydra in cer-
tain ways, but is inclosed in a vase-like formation, the hydrotheca.
3. The latter is a continuation of a tough, membranous
sheath, the perisarc, which covers each part of the whole colony.
Do you notice any modifications of the perisare below the
hydrotheca? Do the modifications serve any purpose?
4. Trace the stem down to the creeping, stolon-like portion
of the colony, the hydrorhiza.
Make a drawing of a colony.
5. The fleshy continuation of the zodid down into the stalk
is termed the cenosarc. Is it in close contact with the perisare?
6. In an expanded hydranth, note the mouth, the arrange-
ment of the tentacles, and the number of tentacles. How is the
OBELIA. 23
individual supported in its cup? Can you trace the celenteric
cavities down through the stalks, and prove them to be continuous
with each other? Motion in the fluid contents of living speci-
mens makes this easy to observe.
7. Examine a hydranth with a high power and look for the
cell-layers characteristic of ccelenterates. Determine how its
tentacles differ from the tentacles of Hydra, and explode nemato-
cysts as with Hydra.
Make a drawing of a hydranth.
8. Look for certain extremities which show neither tentacles
nor any opening in the perisarcal covering. Such a condition
signifies either an undeveloped hydranth or a reproductive
individual. If the latter, it is considerably swollen and is termed
agonangium. ‘The central core or coenosarce of a gonangium, the
blastostyle, should be examined for medusew buds. This may re-
quire a high power.
Make a drawing of a gonangium.
9. You may find free medusze swimming in the dish where
material is kept. If you do, you should examine one, but it
will not prove as satisfactory for study as a larger form, like
Gonionemus, directions for the study of which are given further
over.
In comparing it with Gonionemus notice the small size of
the velum, and the ease with which the bell turns wrong side
out, so the manubrium appears like a handle. At Woods Holl,
Obelia apparently does not always liberate its medusz, and it
is not uncommon to find planule escaping from gonothece.
Frequently those medusze that are liberated have previously
shed their eggs.
10. Study the cellular structure of a hydranth and of a gon-
angium, as seen in cross and longitudinal sections.
Make a drawing of each.
For comparison use any thecate forms, which may be offered
as loose material or as demonstrations, such as Campanularia,
Sertularia, and Plumularia.
24 CCELENTERATA.
PARYPHA.
This form is frequently abundant on the piles of old wharfs,
where the colored colonies form conspicuous masses just below
low-water mark.
Examine the general form of a colony and note, either with
a hand lens or with the naked eye, the stem, or hydrocaulus, as
it arises from the branching, matted hydrorhizal portion of the
colony. The parts of the colony will be seen to differ from the
Leptomedusan (Campanularian) form studied, especially in
branching, rigidity, hydrothece, and gonangia.
Make a drawing to show the formation of the colony.
1. How does a hydranth differ from the hydranth of Obelia
in the matter of tentacles? Is a hydrotheca present?
2. The mouth is terminal and is situated at the end of a
manubrium.
3. The short but rather large body of the hydranth passes
back to the perisarc as the fleshy axis, cenosarc.
4. Notice the medusa buds on the manubrium between the
rows of tentacles. What is their arrangement? This is a form
in which the medusz are not set free, but remain vestigial.
The gonads ripen on the partially developed manubrium of
the medusa. The sexes are separate.
Make a drawing of a hydranth.
5. The arrangement of the attached meduse& is best seen in
sections. In the male meduse numerous spermatozoa will be
found, while the female individuals have a much smaller number
of large ova, which are likely to be in advanced stages of seg-
mentation.
The sections show the same body layers as Hydra, and the
derivation of the medusa as an outpocketing of the wall of the
hydranth is evident.
Make drawings of sections of male and female reprogaeeiae
organs (medusa buds).
For comparison, study Pennaria, Margelis, Hydractinia,
Clava, and Eudendrium.
GONIONEMUS. 25
GONIONEMUS.
As has been seen, the medusze buds are usually produced from
the walls of a specialized hydranth (Leptomedusz) or -from the
manubrium wall of an ordinary hydranth (Anthomeduse). A
series of these buds in various stages would show the formation
of the umbrella-shaped individual with the growth of the marginal
tentacles around the edge. ‘The life-history of this form is not
known, but from its structure we are led to believe that it
belongs to the suborder Leptomedusz.’ It is found in con-
siderable numbers throughout the summer in the border of eel-
grass in the Eel Pond at Woods Holl, where it may be obtained
with adip-net. It is more satisfactory to study than the medusa
of Obelia, as it is much larger and its movements and organs
are more easily observed. In plan of structure the two are
quite similar.
Put a living specimen in a flat-sided jar containing sea-water,
or in a finger-bowl, with a black tile beneath, and notice:
1. Its method of locomotion. To the contraction of what
part of the bell is movement due? How large is the jet of water
that is delivered from the bell? Why is the jet made narrow?
Does the jet necessarily leave at the center or may it be thrown
from one side? Should it be thrown from one side, what would
be the result?
2. Its position in the water when quiet. Why is this position
more desirable than the opposite? With a needle-point prove
that various parts of the body are sensitive.
With either fresh or preserved material notice:
1. Its flattened dome-shape. The convex face is called the
ex-umbrella (aboral), while the concave portion is termed the
sub-umbrella (oral).
2. The velum is the perforated diaphragm that partly closes
in the sub-umbrella. All medusz possessing this structure
are classed as Craspedota. Do you understand its use?
1There is some reason for thinking that the polyp stage of this form
develops directly into the medusa. (See Perkins, Proc. Acad. Nat. Sci.,
Philadelphia, 1902.)
26 CCELENTERATA.
3. In the center of the sub-umbrella is seen the large pen.
dent manubrium, at the extremity of which is a wide-lipped
mouth. If the medusa is alive, feed it with small bits of clam
meat.
4. From the capacious sac at the base of the cavity of the
manubrium, the stomach, the four radiating chymiferous tubes,
or canals, lead to the periphery of the disk, where they open
into the very delicate circular circumferential canal. The four
radii marked out by these canals are called the per-radu. Do
you understand the use of these canals?
5. The gonads hang from beneath the chymiferous tubes into
the sub-umbrellar space. They are lobulated in structure, and
more or less prominent according to maturity and the breeding
season. The eggs or spermatozoa, as the case may be, are de-
hisced from these into the water directly.
6. The tentacles. Is their arrangement a radially symmetrical
one? How are the nematocysts arranged on them? Look for
adhesive organs on them. Of what use are such organs?
Turn your specimen with the velum side toward you and
study the edge of the medusa with a low-power objective for
the sense organs. These are of two kinds:
(a) The larger, round bodies at the bases of the tentacles
communicate with the circumferential canal (which may possibly
be seen along the edge of the bell). They are filled with a layer
of strongly pigmented endoderm cells and are probably light-
perciprent organs.
(b) Other small sessile and transparent outgrowths, situated
between the bases of the tentacles, are the so-called otocysts,
which are probably static organs.
All of the tentacles are abundantly supplied with tactile,
sensory cells. There is a well-established circumvelar nerve
ring (not easily determined in living material) derived from the
ectoderm, also scattering nerve cells beneath the ectoderm in
connection with the muscular tissue. Ex-umbrellar and sub-
umbrellar layers of muscle fibers are also present.
HYDROCORALLINA. SIPHONOPHORA. AURELIA. Ze
Make a drawing from the side, slightly tipped, to show the velum,
and another as seen from the oral surface.
HYDROCORALLINA.
To this group belong forms that have heavy calcareous
exoskeletons. While material is generally not at hand to study
the polyps, it is desirable to study and sketch the characteristic
forms of colonies such as Millepora and Stylaster, and to note
the difference in the distribution of pores. Later you will see
how decidedly these differ from the ordinary stony corals.
SIPHONOPHORA.
Examine living or preserved specimens of Physalia, and
sketch the type with reference to showing, if possible, the follow-
ing structures: (a) pneumatophore, (b) dactylozodids, (c) gastro-
zodids, (d) gonodendrons, (e) tentacles. It will be well to refer
to a text-book to find the positions and functions of each of
these.
SCYPHOZOA.
AURELIA.
This form is one of the common jelly-fishes, and is found
floating freely in the water. It is frequently washed up on shore.
To be appreciated these medusz should be seen as they occur
at the surface of the sea, before they have been handled or in-
jured. Frequently vast numbers may be seen together, all
gently pulsating and thus keeping near the surface. The move-
ment is very different from that of most hydrozoan medusa,
being very deliberate and graceful.
If living material is offered, study the method of locomotion
and compare it with the locomotion of Gonionemus. Like the
latter, the discoid animal presents ex-umbrellar (aboral) and sub-
umbrellar (oral) surfaces, but the edges of the disk are indented,
fringed with very numerous short tentacles, and a velum is
wanting. What difference does the velum make in locomotion?
The ex-umbrellar surface presents little of interest. In the
28 CCELENTERATA.
live specimens, however, prove that the animal is sensitive over
this area as elsewhere.
Preserved and hardened material is better than living for the
study of the rest of the anatomy of this form. With a specimen
in water in a finger-bowl, with a black tile for the background,
find the following from the sub-umbrellar surface:
1. The shape of the animal. Is the margin perfectly circular
or regularly indented? Are all of the marginal portions similar?
2. Four large, fringed oral arms or lips hang from the corners
of the nearly square mouth, which is located in the center. No-
tice how each arm is similar to a long, narrow leaf, with the
sides folded especially along their margins. Examine the arms
for nematocysts. Do you understand how the animal gets its
food? If the arm edges appear to be covered with dark specks
and granules, examine to see if embryos may not be entangled.
3. The mouth is found to lead by a short gullet into a rather
spacious stomach, which is produced in the region between each
two corners of the mouth to form a gastric pouch. Determine
the shape of the stomach.
4. The remaining parts of the digestive (and also in this
case circulatory) system include the numerous radial canals
and the single circumferential canal.
(a) Directly beneath each oral arm a per-radial canal is given
off, which, at a short distance from the stomach, gives off a
branch on either side. The per-radial canal proper usually
continues straight to the marginal circumferential canal, without
further subdivision, but the two side branches above mentioned
in turn subdivide several times.
(6) From the peripheral wall of each gastric Be three
canals pass toward the margin; the middle one (inter-radial
canal) branches somewhat after the manner of the per-radial
canals, but the other two (ad-radial canals) continue to the
circular canal without further branching.’
1In most cases the foregoing canals are very evident, but if they are
not, they may be injected w ‘ith water in which carmine is mixed, by insert-
ing a large-mouthed pipet into the stomach.
AURELIA. 29
5. The position of the gastric pouches is made clearly mani-
fest by the gonads, which lie on the floor of the pouches, as frill-
like structures, horseshoe-shaped, with their open sides toward
the mouth. The ova or spermatozoa are shed into the stomach and
pass out of the mouth. Embryos in various stages of develop-
ment may frequently be found adhering to the oral arms. The
sexes are separate. On the sub-umbrellar surface, opposite
each gonad, is a little pocket, the sub-genital pit, which opens
freely to the outside. Whatever purpose this may serve, it
does not function to conduct the genital products to the outside.
6. Parallel with the inner or concave border of each gonad
is a row of delicate gastric filaments. These are supplied with
stinging cells, and they may aid in killing live food taken into
the stomach. These structures are not present in the Hydro-
zoan medusa.
7. At the marginal extremity of each per-radial and inter-
radial canal there is an incision on the edge of the animal, in
which there are sensory organs. In each incision find:
(a) A tentaculocyst in the form of a short, club-like struc-
ture containing a prolongation of the circular canal. At its
outer extremity are calcareous concretions or lithites, and a pig-
ment-spot or ocellus. Each tentaculocyst is protected aborally
by a hood-like projection, and on the sides by marginal lappets.
(b) Two depressions, one above and the other below the
tentaculocyst. These have been assigned olfactory functions,
and are called the oljactory pits.
Make a drawing showing the profile of the entire animal, and
show the structure of at least one quadrant, as seen from the oral
surface.
If time permits study a developmental stage, “‘ephyra,” and
compare it with the adult.
By way of comparison, examine demonstrations of Cyanea,
Dactylometra, Lucernaria, or other forms belonging to this group.
30 CGELENTERATA.
ACTINOZOA.
METRIDIUM. (Sea-Anemone.)
Specimens are quite common on piles, as well as on rocky
bottoms, and may be easily observed by means of a glass-
bottomed pail. Most of the observations can be made much
better on specimens in aquaria, but it is desirable to see their
natural surroundings.
1. Notice the shape and attachment of expanded, living speci-
mens in an aquarium, or in a deep finger-bowl. The free end,
called the disk or peristome, is fringed with tentacles, and the
elongated mouth is located in the middle of this area. At one
or both angles of the mouth the lips are thickened into what is
called a stphonoglyph.
Make a drawing of the animal.
2. Feed a specimen with bits of mashed clam to ascertain
its manner of taking in food. Drop bits on the tentacles at one
time, and disk at another.
Endeavor also to determine whether there are currents
constantly passing in or out of the mouth that are due to-ciliary
action.
3. Irritate the animal and observe its manner of contraction.
When fully contracted, if the irritation is continued, thread-
like structures, acontia, are thrust out through minute pores,
cinclides, in the body-wall.
Make a drawing of the contracted animal.
Internal Anatomy.—Using preserved material, place the edge
of a razor across the peristomial area, at right angles to the
mouth-slit, and divide the animal from disk to base into halves.
1. Note the extent of the esophagus and siphonoglyphes;
they lead into the celenteric chamber. Find the extent of this
chamber, and the method of its subdivision by delicate parti-
tions, the mesenteries, or septa. Are all of the mesenteries alike?
2. Forming the free edges of the mesenteries, below the
esophagus, are the convoluted mesenteric filaments, which are
METRIDIUM. 31
secretory organs that are probably equivalent to the gastric
filaments of the Seyphozoa.
3. Quite near the bases of the mesenteries are the attachments
of the acontia. What relation have they to the mesenteric fila-
ments?
4. Also located on the mesenteries, and arranged parallel
to the filaments, but back from the edge a bit, are the repro-
ductive organs or gonads. Are they found on all of the mesen-
teries? The ova or spermatozoa are shed into the ccelenteric
chamber and pass out through the mouth.
Cut one of the halves of your specimen transversely in the
region of the esophagus, and study the arrangements of the
mesenteries, their attachments, ete.
5. How many pairs of primary mesenteries, i. e., those attached
both to the outer body-wall and to the esophagus, are there?
The directive septa are those at the angles of the esophageal tube.
The portion of the ccelenteric cavity between any two pairs
of mesenteries is termed an inter-radial chamber. The space
between the two mesenteries of each pair is called an intra-
radial chamber.
6. Carefully determine the disposition of the longitudinal
retractor muscles on the mesenteries. Do they occupy similar
positions on all of the mesenteries ?
7. Examine the upper parts of the mesenteries for openings,
septal stomata, that put the chambers in communication
8. Are the tentacles solid or hollow?
Make a drawing oj a longitudinal section and another of a
cross-section. Put into these all of the points of the anatomy you
have seen.
If time and opportunity permit, it is very desirable that this
form should be compared with specimens of the order Madre-
poraria, and later with the Alcyonaria. Such a form as Astran-
gia may easily be obtained either alive or properly preserved,
and will serve to show the relation of the hard parts of the coral
_ to the polyp. You should understand the relation of the septa
32 CCELENTERATA.
and the mesenteries, and of the polyps to each other. If speci-
mens are at hand, compare such forms as Orbicella, Favia, and
Meandrina, or any forms that show gradations from separate
calices to fused groups, and understand the positions of mouths,
the arrangement of the ccelenteric chambers, and the way in
which the colony has come to its present form. You should
also examine large branching colonies and determine why branches
are formed and how they arise.
Examine the structure of an Alcyonarian colony and see
how the polyps are placed. The structure of the expanded
polyps is nicely shown by Renilla. The spicules of such forms
as Gorgonia may be obtained by boiling a portion of a colony
in caustic potash. What purpose do such spicules serve?
CTENOPHORA.
| MNEMIOPSIS.
This form belongs to the group of animals popularly called
‘“‘comb-jellies,”’ and occurs along the coast in irregular abun-
dance during the summer months. Specimens are very phos-
phorescent when disturbed, so, when they are abundant, the
display caused by them while rowing at night is sometimes bril-
liant. They may frequently be seen during the daytime and can
often be satisfactorily observed in the shade of a wharf when the
water is calm.
Unmutilated, living material can be studied to best advan-
tage, but preserved material may be had that is quite satisfac-
tory for anatomic study.
1. In general appearance a specimen resembles a hydrozoan
medusa, with its aboral surface elongated until, as a whole, it
approaches the shape of a fowl’s egg.
2. The broader or oral end bears two heavy terminal lobes,
between the bases of which is the slit-like mouth. We may con-
sider the elongation of the mouth to be in the antero-posterior
line.
-~ ee ee eee eed
feat So ae eee te
MES SU Se Se Se ne SS
Settay yr ee SSS
err cce tise sben sepeeeseeeenst eas ba geet st
SEIS See os
secphecemichstestearetsrets yiijge eee : Sseee
ae > Mists eR ES ee
32 RES ee ee
Bao Ss el eee pawn be pwecen wp hepeatas se ssee awe, oppress semeeeete
Ge wernt ate cp ae we eer eee mre me tome hee eereb bree a eeee eee
333 at er to bo bas probe rege ara ene ere rst -) Peheer ema e thre mesos son. ee ee ee ere
SSS ooops wore ee ns bape le See ween ee renee probe eos buena as yee os ter omen
ins ss SEs aS ss: Poorman se Sate sces sere po we Pe Se PS nae STS SS SET s OOPS OE OS SSPE we or oe waders cate Fe ow ree OS re hase Gees ese= em ereeeore:
See a mt pel oe bn Oe Pe een oe Renn ae en ee ee ee ee ee ee eee ene ees : ete wace tee
TIS le cae = es Ten pees beer pos eee ees
eS I tap ise
oR oss Tt Se SSFP TSS
a a pe ae ee oe >
ee ee eee to te ekesetes
eS Se ee eee
so oa re Om on pn ae ne a oe He om
a
pers eesa ee teers tee
ee ee ire eee a eet ee
ioerrteteeetrttictpesssectessarn street ers Oe yo tare epee neo here eiivactarsttieted <2 Sie ee
Mates Ss et er oe Me eee :
ea ee pe me me er
se ee
re etrrt sree
5 ope et eete se oe
ei tet ne ea we ee ee ee ene
Se en +e RE OE tm OO ne wm ome
STE Te a a a EO
yep ee Se ee eo
ee beaess Mate
it Tha
ee ee een ee
eos
i cigtetsspee este os) Ske ees
Pye ee SL ee et Pe e= we
eee knee ee S Mee pee ee
eq teatyessperagien s s syatgtsterpieseey py eens o> Aen eye een sem
pies Lowe te ale wheter a iaedetilipthte ketene! nebo 33-44 ocean ne
ae be Bee ae Oo a erent ee eee rate ee
Ne 0 Oe em tee me oe .
= Bits toere esta
we See ene = Paes 7 =
eae Se ae ee ee
Serenata rae =
eos ewer e— bo ee>
ye ene =)
2a pene en
a
soe me ee ga pe pe ep e mek ee te mee pl ee eee roe tS SSE Sa en St Sper y— ot
ep oe ee ere a a eee ce Le es te oe ole pw he Pees Oe ime bola ee oe ee es oe ee a Te ie pe ee Ba ee eh ee nn ee
Oe SS) ae Ba pe raya ee OE peta eke we Pepe Nee eee te wee ats
eppaenpuse sip leh) Hereeesr see tl, ee at re pe ae Bee ee eee os ee ee
Itt Sse —
bhp ope moro pe ace faa aS isa Spel errnge peer sper
preresyarbestee taf. cys Siiaeteestes
owners Spee eee se
SS ee ats
SSS reon> wee ne ape ee ee
Fm re ok = pe pepe eye
SS ee ee
verey es 4 oo =
Tee ee Se SA
- ~ a reset pa be
Sereeteisreete Sos aera ete yeas
Ca Seah ikea etd aoe
ne eR ee eee
-o- oe SI SESte Tt:
+ 7 Ne ae ee eee ge le ete oe me
ke ee pe
PF a a PS ee pe haoere ee am
ome” € Starx = Se
ph ees a ee en
ngs as FS eee ee SS
To BS
™ Se
=e >= pape
rere er 3752332
ane aseserp saber pees re el ere ma te mene ipks Sota tne otaaetenod tn oe oe eee one. bee
aoe: SEetess Ssieo toc Ser eti cert bret of
ae ae ene
= ot ee eae
=e eee ee
So nr a eer ae ee gees
Ieee
Se ee
ae aye ey 2 Oe re
ae att eins ee heehee ae ee es
=)
Se
Bi Peg iisttssseee steers
oe ee
ee
oe he on
iS Pe bs Be bey wm awebe a
SS eee
pee Seale LS ee
2 SS
pez.
ee ere =
=i a eee
i
i
'
hy
stl
.
or
|
“:
.
rryerreee
Wis
bits!
|
rertt
ty
rininst
beer
{
elt ft
int
eitit
iy
4
rh
.
bets
wd
Hd