Thesis (MSc)--Stellenbosch University, 2001. / ENGLISH ABSTRACT: PELOMEDUSA SUBRUFA is a freshwater turtle widely distributed throughout Africa and Madagascar, and is described as a Tropical to Sub-tropical species. 1 examined the female and male reproductive cycles of P. subrufa, over a 20-month period to determine whether they display a typical Tropical to Sub-tropical type reproductive cycle (pre-nuptial) or a typical Temperate Zone type reproductive cycle (post-nuptial). Blood and tissue samples were collected from wild specimens captured in the Western Cape, South Africa and these samples were supplemented by tissue samples obtained from museum specimens. In female P. subrufa seasonal variation in related circulating reproductive hormones in the plasma (estradiol, progesterone, and testosterone) were analyzed using validated ELISA kits. Plasma vitellogenin (yolk precursor produced in liver) was measured using a newly developed universal vitellogenin ELISA for vertebrates (UNIVTG). Ovarian follicles were measured (± 0.1 mm) and female ovaries were staged macroscopically (non-active, pre-vitellogenic, vitellogenic, gravid), and results were confirmed via histological sectioning of ovaries and oviducts. Females exhibited a cyclic reproductive pattern, with distinct phases of follicular enlargement (vitellogenesis), ovulation and a gravid period. Seasonal timing of the reproductive cycle coincided with those of other temperate zone freshwater turtles. Vitellogenic recrudescence began in summer (late December), and continued unabated through winter with ovulation occurring in the following spring (September-October). My data suggested that P. subrufa females mostly lay a single clutch of eggs during the late-spnng summer period (September through January). Clutch size varied between 7 -3 7 eggs, with the number of eggs being significantly correlated with maternal body size (r = 0.82, P < 0 001). Plasma estradiol and plasma vitellogenin concentrations peaked once during the ovarian cycle, typically coinciding with the period of early- to mid-vitellogenesis in late summer. Plasma testosterone varied throughout the year, but significant increases were measured during the ovulation and mating period in spring. Plasma progesterone concentrations were significantly elevated during the gestation period prior to ovi-position in mid-summer (December). In male P. subrufa spermatogenesis in mature specimens was distinctly seasonal and timing of the reproductive cycle coincided with those of other temperate zone freshwater turtles. Spermatogemc recrudescence began in summer, following emergence from a winter hibernation period (brumation) and spring mating. Peak testicular volume and maximum spermiogemc activity occurred in late summer and early autumn. Testicular regression commenced in autumn through winter. Spermatozoa were abundant in the ducti epididymi throughout the year. Plasma testosterone concentrations peaked once during the testicular cycle, typically coinciding with spermio genes is in late summer, early autumn. Ducti epididymi diameter showed significant variation throughout the year, whereas the epithelial cell height showed no significant seasonal variation. Peak secretory activity coincided with spermiogemc activity and high circulating testosterone concentrations in late summer, early autumn. Testicular recrudescence was correlated with increasing ambient air temperatures, photopenod and summer rainfall, whereas testicular regression, during late autumn, corresponded conversely with decreasing ambient air temperatures, photopenod and rainfall. Female and male reproductive cycles were asynchronous in that the peak spermatogenic activity occurred in autumn at the time when most females were depositing yolk in growing ovarian follicles. Therefore, adult females displayed a typical postnuptial vitellogemc cycle and adult males displayed a typical post-nuptial spermatogenic cycle. Differences between sexes in body size are common in many animals, and the African helmeted turtle is no exception. Sexual size dimorphism (SSD) in P. subrufa was pronounced, and using principal component analysis, it was clear that adult male P. subrufa was significantly larger than adult females. Using carapace length as the measure of body size (covariate), adult males, adult females, and juveniles differed significantly in absolute size of the carapace width, carapace depth, plastron length, plastron width, and head depth. However, there was no significant difference between adult males, adult females and juveniles in head width and head length. Therefore, adult males were larger than adult females in the seven traits measured, except in carapace depth where the females were significantly larger In the occurrence of ontogenetic growth patterns, the adults grow at a slower rate than juveniles in plastron length. There was no significant difference between adults and juveniles in shell width, however in depth, the adults grow at a faster rate when compared to the juveniles. Adults significantly grow at a faster rate than juveniles in absolute head size as well. However, when these traits were used as a whole data set (eight traits measured), there was no difference in growth rate between adults of either sex. Similarly, there was no significant difference in adults compared to juveniles in shell size, however, adults grow at a faster rate than juveniles in absolute body size and head size. Differences in body size, and in the size of traits such as shell measurements and head measurements relative to absolute body size, were assessed to clarify SSD of P. subrufa in South Africa. / AFRIKAANSE OPSOMMING: PELOMEDUSA SUBRUFA is ‘n varswaterskilpad wat wyd verspreid oor Afrika en Madagascar voorkom en word beskryf as ‘n Tropiese tot Sub-tropiese spesies. Die manlike en vroulike voortplantingspatroon van P. subrufa is oor ‘n tydperk van 20 maande bestudeer om vas te stel of hul voortplanting ooreenstem met ‘n tipiese tropiesie tot sub-tropiese voortplantingspatroon of ‘n tipiese gematigde-sone voortplantingspatroon. Waterskilpaaie is uit damme in die Wes-Kaap, Suid-Afnka gevang en bloed- en weefselmonsters is versamel. Materiaal en data is aangevul deur weefselmonsters van waterskilpaaie wat in museumversamelings gehuisves word. Ovarium follikels in P. subrufa wyfies is gemeet en die wyfies se ovanums is makroskopies gegradeer (onaktief, pre-vitellogenies, vitellogemes, dragtig) en resultate is deur histologiese snitte van die ovaria en ovidukte bevestig. Wyfies vertoon ‘n sikliese voortplantingspatroon, met duidelike fases van follikulere groei (vetllogenese), ovulasie en dragtigheid. Sirkulerende voortplantingshormone in die bloedplasma (estradiol, progesteroon en testosteroon) is ook geanaliseer met behulp van gevalideerde hormoonspesifieke ELISA bepalings. Plasma vitellogeen (‘n dooiervoorloper wat in die lewer vervaardig word) konsentrasies is ook bepaal met ‘n nuut ontwikkelde, universele (spesifiek vir werweldiere) vitellogeen ELISA (UNIVTG). Seisoenale tydsberekemng van die voortplantingsiklus het ooreengestem met die van ander varswaterskilpaaie vanuit die Gemagtigde-sone. Vitellogenese het in die somer begin en duur voort deur die grootste gedeelte van die somer, herfs en winter gevolg deur ovulasie in die daaropvolgende lente (September - Oktober). Die data ingewin stel voor d a t/5. subrufa wyfies meestal een broeisel eiers tydens laat lente-somer le (September tot Januane). Broeiselgrootte het gewissel tussen 7-37 eiers, met die hoeveelheid eiers wat beduidend met moederlike liggaamsgrootte gekorreleer was (r = 0.82, P < 0.001). Plasma estradiol en vitellogeen konsentrasies het een keer tydens die ovariumsiklus gepiek, en gewoonlik saamgeval met vroee tot middel vitellogenese in die laat somer. Plasma testosteroon het dwarsdeur die jaar gevarieer, maar beduidende toenames is gemeet tydens ovulasie en die paartydperk in die lente. Plasma progesteroon konsentrasies was beduidend hoer tydens dragtigheid kort voor eierlegging in die middel van die somer (Desember). In volwasse P. subrufa mannetjies was spermatogenese sterk seisoenaal en het die voortplantingsiklus ooreengestem met die van ander varswaterskilpadspesies wat in die gematigde streke voorkom. Na ‘n oorwinteringsperiode (brumasie), volg die panngstydperk gedurende die lente. ‘n Nuwe spermatogemese siklus het in die somer begin. Maksimale spermatogeniese aktiwiteit en testis-volume word in die laat somer en vroee herfs bereik. Testikulere regressie neem in aanvang in die herfs en duur voort tot na paringstyd in die lente. Tydens testikulere regressie word spermatosoe in die ducti epididymi gestoor. Plasma testosteroon konsentrasies het in die laat somer en vroee herfs gedurende die testikulere siklus, spermiogenese (sperm produksie fase), gepiek. Die grootte (omtrek) van die ducti epididymi het beduidende vanasie dwarsdeur die jaar getoon, terwyl epiteel selhoogtes geen beduidende seisoenale vanasie getoon het nie. Piek sekretonese aktiwiteit het saamgeval met spermiogeniese aktiwiteit en hoe vlakke van sirkulerende testosteroon tydens laat somer en vroee herfs. Testikulere groei het goed gekorreleer met toenemende omgewingstemperatuur, fotopenode en reenval, terwyl testikulere regressie in herfs met ‘n daling in omgewingstemperature, fotopenode en reenval gekorrespondeer het. Die vroulike en manlike voortplantingspatrone was nie goed ge-sinkroniseerd nie, deurdat piek spermatogeniese aktiwiteit tydens herfs voorgekom het, gedurende die tyd waann meeste wyfies besig was om dooier in groeiende ovarium follikels neer te le. Daarteenoor vertoon die mannetjies testikulere regressie tydens die pre-ovulatoriese fase en ovulasie penode van die wyfies. Dus toon volwasse wyfies ‘n tipiese gematigde sone vitellogeniese patron en volwasse mannetjies ‘n tipiese gematigde sone spermatogeniese patroon. ‘n Verskil in liggaamsgrootte tussen die geslagte is 'n algemene verskynsel by baie diere en P. subrufa is geen uitsondering me. Daar was wesenlike geslagtelike grootteverskille (SSD) in P. subrufa en ‘n hoofkomponent analise (PC A) het getoon dat daar beduidende morfometnese verskille tussen volwasse mannetjies en wyfies was. Deur karapakslengte as 'n maatstaf vir liggaamsgrootte te gebruik (mede-veranderlike), het volwasse mannetjies, volwasse wyfies en onvolwassenes beduidend verskil ten opsigte van absolute grootte van hul karapaksbreedte, karapaksdiepte, plastronlengte, plastronbreedte en kopdiepte. Geen beduidende verskil in kopbreedte en koplengte in volwasse mannetjies, volwasse wyfies en onvolwassenes is gevind nie. Derhalwe was volwasse mannetjies groter as volwasse wyfies in sewe van die liggaamseienskappe wat gemeet is, buiten vir karapaksdiepte waar die wyfies beduidend groter was. In terme van die voorkoms van ontogenetiese groeipatrone het volwassenes teen ‘n stadiger tempo as onvolwassenes in plastronlengte toegeneem. Daar was geen beduidende verskil in die groeitempo van dopbreedte tussen volwassenes en onvolwassenes nie, alhoewel dopdiepte van volwassenes teen ‘n vinniger tempo gegroei het as die van onvolwassenes. Absolute kopgrootte van volwassenes het ook teen ‘n vinniger tempo gegroei as in onvolwassenes. Wanneer hierdie eienskappe as ‘n volledige datastel gebruik word (al agt gemete eienskappe), wil dit voorkom asof daar geen verskil in groeitempo van volwassenes van die onderskeidelike geslagte is me. Daar was geen beduidende verskil tussen volwassenes en onvolwassenes, ten opsigte van dopgrootte nie, alhoewel volwassenes klaarblyklik teen ‘n vinniger tempo in liggaamsgrootte en kopgrootte toeneem. Verskille in liggaamsgrootte en grootte van ander veranderlikes, soos byvoorbeeld dop- en kop-eienskappe, relatief tot absolute liggaamsgrootte, word aangebied om geslagsdimorfisme in P. subrufa vir die eerste keer te beskryf.