Tripanossomas de répteis há muito são descritos em lagartos, tartarugas, crocodilos e cobras de todos os continentes. A classificação destes tripanossomas tem sido feita de acordo com a morfologia de formas sanguíneas e hospedeiro de origem. No entanto, estes critérios tradicionais não são suficientes para descrição de espécies, gerando varias sinonímias. Vinte e sete isolados foram mantidos em culturas contínuas. Estas são as primeiras culturas de tripanossomas de lagartos da América do Sul, e as unicas culturas de tripanossomas de jacarés e cobras disponíveis no mundo. As analises filogenéticas de tripanossomas de répteis realizadas usando seqüências dos genes SSUrRNA e gGAPDH corroboram a polifília de tripanossomas de répteis. O clado que inclui isolados provenientes de cobras e lagartos sugere uma associação com hospedeiros Squamata, sugerindo que flebotomíneos tenham um papel importante na transmissão destes tripanossomas. Os dados morfológicos e moleculares resultantes deste estudo permitiram a descrição de 7 novas espécies de tripanossomas. / Reptile trypanosomes have been described in lizards, tortoises, crocodiles and snakes of all continents. Reliable classification of reptile trypanosomes requires analysis based on traditional parameters and molecular phylogeny. However, these criteria are not enough for species description, generating synonymies Twenty-seven isolates were established in continuous cultures: 9 from lizards, 8 from snakes and 10 from caimans. Cultures of these isolates displayed distinct morphology and growth behavior, representing the first cultures of lizard trypanosomes from S. America and the only available cultures from caimans and snakes. The phylogenetic analysis of the reptile trypanosome using SSU rRNA and gGAPDH gene sequences corroborated the polyphyly of reptile trypanosomes. The clade harboring snake and lizard trypanosomes suggested an association with Squamata hosts, with evidence that sand flies play an important role in the transmission of these trypanosomes. Morphological and molecular data from this study permitted the description of 7 new trypanosome species.

abstract: Traumatic injury to the central nervous or musculoskeletal system in traditional amniote models, such as mouse and chicken, is permanent with long-term physiological and functional effects. However, among amniotes, the ability to regrow complex, multi-tissue structures is unique to non-avian reptiles. Structural regeneration is extensively studied in lizards, with most species able to regrow a functional tail. The lizard regenerated tail includes the spinal cord, cartilage, de novo muscle, vasculature, and skin, and unlike mammals, these tissues can be replaced in lizards as adults. These studies focus on the events that occur before and after the tail regrowth phase, identifying conserved mechanisms that enable functional tail regeneration in the green anole lizard, Anolis carolinensis. An examination of coordinated interactions between peripheral nerves, Schwann cells, and skeletal muscle reveal that reformation of the lizard neuromuscular system is dependent upon developmental programs as well as those unique to the adult during late stages of regeneration. On the other hand, transcriptomic analysis of the early injury response identified many immunoregulatory genes that may be essential for inhibiting fibrosis and initiating regenerative programs. Lastly, an anatomical and histological study of regrown alligator tails reveal that regenerative capacity varies between different reptile groups, providing comparative opportunities within amniotes and across vertebrates. In order to identify mechanisms that limit regeneration, these cross-species analyses will be critical. Taken together, these studies serve as a foundation for future experimental work that will reveal the interplay between reparative and regenerative mechanisms in adult amniotes with translational implications for medical therapies. / Dissertation/Thesis / Differentially Expressed Genes in the Early Regenerating Lizard Tail / Gene Ontology of Differentially Expressed Genes in the Early Regenerating Lizard Tail / KEGG and Reactome Pathway Analysis of Differentially Expressed Genes in the Early Regenerating Lizard Tail / 3D Reconstruction of an Alligator Regenerated Endoskeleton / Lateral 2D Serial Sections of a Regenerated Alligator Tail / Doctoral Dissertation Biology 2020

This study determined the passage time and phage propagation time of salmonella specific phages, Felix O1 and S16, in 10 bearded dragons, based on re‐isolation from cloacal swabs and faecal samples following oral administration, as a possible tool for reducing salmonella shedding. In Study 1, Felix O1 was administered orally for 12 consecutive days. Over 60 days, swabs were taken from the oral cavity and cloaca and qualitative Salmonella detection as well as salmonella quantification from faecal samples were performed. In Study 2, a phage cocktail (Felix O1 and S16) was administered to half of the tested animals. Salmonella (S.) Eastbourne was also given orally to all animals. Oral and cloacal swabs were tested as in Study 1, and faecal samples were collected for phage quantification. Various Salmonella serovars were detectable at the beginning of the study. The numbers of serovars detected declined over the course of the study. S. Kisarawe was most commonly detected. Salmonella titres ranged from 102 to 107 cfu/g faeces. The phages (Felix O1 and S16) were detectable for up to 20 days after the last administration. The initial phage titres ranged from 103 to 107 pfu/ml. The study shows that the phages were able to replicate in the intestine, and were shed for a prolonged period and therefore could contribute to a reduction of Salmonella shedding.

In 1926, the German biologist Johanna (Hanni) Hrabowski published a study of the morphology and development of the fetal placenta in lizards that has proven to be of historical importance. Her anatomical descriptions and interpretations identified developmental patterns that differ from other amniotes – features now recognized as unique attributes of squamate (lizards and snakes) development. Her 1926 monograph presented the first histological comparison of fetal membranes in closely-related oviparous and viviparous reptiles, thereby establishing a comparative framework for understanding placental specializations for viviparity. Hrabowski reported that yolk sac development did not differ between oviparous and viviparous species. The novel, shared components of yolk sac development she identified are now recognized as the foundation for the unique yolk sac placenta of reptiles, the omphaloplacenta. In addition, Hrabowski's extensive ontogenetic sampling and the detail and accuracy of her anatomical descriptions set high standards for subsequent studies of comparative evolutionary embryology.

To gain a better understanding of the role of steroid hormones in vertebrate reproduction, we quantified steroid hormone concentrations in a free ranging population of the Northern Pacific rattlesnake, Crotalus oreganus. Plasma steroid hormone concentrations were quantified for both male and female snakes throughout the active season (Mar-Oct). We measured testosterone (T), 5α-dihydrotestosterone (DHT), and corticosterone (B) concentrations in male and female snakes. 17β-estradiol (E2) and progesterone (P) were measured in females only. We also observed breeding behaviors (e.g. consortship, courtship, and copulation) in the field and measured testis and follicle size in male and female snakes from museum collections. Our results indicate that C. oreganus in central California utilizes a bimodal pattern of breeding, with mating and agonistic behavior occurring in the spring and the late summer/fall. Each breeding season corresponds with elevated or highly variable androgen (T and DHT) levels. Several female snakes had high E2 concentrations in the spring and fall, coincident with vitellogenesis and mating. Females with high E2 concentrations also had high T and DHT concentrations. Corticosterone concentrations in males are not related to either time of year or concentrations of any other hormones quantified. This suggests that the breeding season in this population may not demand a significant increase in energy mobilization by glucocorticoids. Measurements of testis volume show that testes are regressed in the spring when the majority of breeding was observed in this population and reach peak volume in August and September during spermatogenesis. Multiple regression analyses revealed that in female snakes, P is positively correlated with T and DHT, and E2 is correlated with T. Since these results are strictly descriptive, experimental studies are needed to identify the functional significance of these results.

This project evaluated the influence of habitat parameters and distance from low-order streams on herpetofauna communities, in addition to evaluating the efficacy of 2 sampling methods. Amphibians were associated with mature hardwood forests with high density of large, highly decayed woody debris. Reptiles were associated with mixed pine-hardwood forests, woody debris, and vertical canopy structure. Overall, woody debris was an important habitat feature for amphibians and reptiles. Amphibians were detected in greater abundance and species richness near the stream. Reptiles were similar in abundance and species richness as distance increased from low-order streams. I recommend a SMZ width ¡Ý50 m on each side of the low-order streams in east-central Mississippi. Area-constrained and funnel-pitfall traps each detected species that the other method was unable to sample due to inherent biases. I recommend the use of both survey methods when investigating the entire terrestrial herpetofauna community in mature, riparian and upland forests.

I conducted surveys of amphibian, reptile, and small mammal communities surrounding 4 isolated, upland and 6 stream-connected temporary wetlands on Tombigbee National Forest and Noxubee National Wildlife Refuge in north Mississippi from May 2001 - March 2006. Trap captures yielded 17 amphibian species (n = 11,142), 21 reptile species (n = 541), and 10 small mammal species (n = 472). Upland pools supported greater diversity of Ambystomatid salamanders, anurans, lizards, and mice (Peromyscus spp.), than floodplain pools. Factors including landscape position of pools, proximity to alternate water sources, and barriers to dispersal potentially influenced faunal communities of temporary wetlands. Infrared-triggered cameras were used to monitor mammalian activity surrounding ephemeral wetlands to determine potential depredation of pitfall traps. Raccoons (Procyon lotor) accounted for most images (35% total images). I assessed relative effectiveness of sampling techniques to capture and retain herpetiles in presence of depredation. I tested pitfall trap, funnel trap, and pitfall trap combined with exclusion cover along driftence arrays. Overall, pitfall traps of both designs produced more captures than funnels, and excluded-pitfalls yielded greater captures than un-excluded pitfalls for most amphibians. Overall mortality rates were <2% of total captures with anurans accounting for most (63.30%) mortality. I submit that in long-term studies, pitfall traps with exclusion are prudent to limit sampling bias and mortality occurring with depredation of captured herpetofauna. During trapping, pit-traps of both designs yielded incidental captures of small mammals. Capture rates for small mammals were similar in un-excluded pitfall traps and excluded pitfall traps. Southern short-tailed shrews (Blarina carolinensis) and mice accounted for 93% of total captures and suffered 76% and 52% mortality, respectively, potentially due to exposure, starvation, flooding, and/or depredation. Additionally, one species listed as rare in Mississippi was captured during herpetofaunal surveys, oldfield mouse (Peromyscus polionotus). Due to potential impact of pitfall trapping on small mammals, I recommend that researchers either alter trapping methods to address non-target hazards (frequency of checking traps, providing shelter) or work cooperatively using an integrated survey approach for herpetiles and small mammals to limit trap mortality.

While substrate-borne vibrations are utilized by different reptile species, true conspecific communication via biotremors has not yet been demonstrated in reptiles. This study follows a preliminary report that the veiled chameleon (Chamaeleo calyptratus) could produce biotremors in communicative contexts. I tested chameleon behavioral sensitivity to vibrations by placing them on a dowel attached to a shaker emitting vibrations of 25, 50, 150, 300, and 600 Hz and then measured their changes in velocity before and after the stimulus. I then paired chameleons in various social contexts [anthropogenic disturbance (human disruption of animal); dominance (malemale; female-female C. calyptratus); courtship (male-female C. calyptratus); heterospecific (C. calyptratus + C. gracilis); and predator-prey (adult + juvenile C. calyptratus)] and used a video camera and accelerometers to record their behavior. This study demonstrates that chameleons produce biotremors and that receivers exhibit a freeze response when exposed to a simulated biotremor stimulus. Furthermore, veiled chameleons produce biotremors in anthropogenic disturbance, conspecific dominance and courtship contexts, and these biotremors are elicited by visual contact with another adult conspecific and heterospecifics. Overall, two classes of biotremor were identified, "hoots” and “rumbles,” which differ significantly in dominant frequency and waveform. No correlation was identified between animal size and dominant frequency of the biotremors they produced as biotremors originate from rapid muscle contractions. Juvenile chameleons of two months of age are able to produce biotremors, suggesting this behavior may have multiple functions. Overall, the data suggest that the veiled chameleon has the potential to utilize substrate-borne vibrational communication during conspecific and possibly heterospecific interactions.

Artificial retreats are increasingly used to sample animal populations and in attempts to boost animal numbers in degraded habitats. Here, I test potential applications of artificial retreats for lizards inhabiting a coastal environment of high conservation value (Kaitorete Spit, New Zealand). I first conducted a pitfall-trapping survey examining the distribution and relative abundance of lizards in duneland, farmland and shrubland habitats, and tested the influence of trap placement on capture rates. Capture rates of the diurnal skinks Oligosoma maccanni (McCann�s skink) and O. nigriplantare polychroma (common skink) were highest in duneland and farmland, respectively, and were most sensitive to the distance separating traps from the nearest cover (the greater the distance, the lower the capture rate). Captures of O. lineoocellatum (spotted skink) and Hoplodactylus maculatus (common gecko) were rare. Secondly, in separate chapters I test the utility of artificial retreats for monitoring: 1) a preference trial examining relative use of three types of artificial retreats by skinks (O. maccanni and O. n. polychroma) and geckos (H. maculatus); 2) a comparison of the effectiveness of artificial retreats relative to pitfall traps for detecting cryptic and primarily nocturnal geckos (H. maculatus) following translocation; and 3) capture-recapture estimation of population parameters (survival and abundance) of H. maculatus. I found that: 1) geckos strongly preferred retreats made of Onduline over corrugated iron and concrete tiles, whereas skinks exhibited no apparent preferences; 2) artificial retreats were more effective than pitfall traps for detecting geckos following translocation; and 3) monthly survival and recapture probabilities of geckos varied with age-class and over time. Estimated survival was unexpectedly low, possibly due to excessive trap spacing. I developed a new capture-recapture model specifically for population size estimation with data from artificial retreats, which gave estimates that were up to 50% greater than those predicted by conventional capture-recapture models. I caution that permanent placement of artificial retreats in long-term studies may be inappropriate for estimation of population parameters due to potential habitat-enhancement effects and/or altered predation risk. Thirdly, I conducted a capture-recapture field experiment, using a replicated Before-After-Control-impact (BACI) design, to test the relative effects of habitat manipulation (artificial retreat addition) and partial predator removal (by fencing) on annual survival of duneland skink (O. maccanni) populations. Survival increased at sites with predator exclosures, but not at control sites or following the addition of artificial retreats, either alone or in combination with a predator exclosure. The magnitude of the increase in survival for the exclosure-only treatment was small, but sufficient to change the trajectory of an apparently stable population into an increasing one, suggesting that the population is limited by predators. Predator control, but not the addition of artificial retreats, is predicted to benefit O. maccanni. To conclude, the Onduline design developed here appears to be particularly useful for sampling cryptic, terrestrial geckos; however, artificial retreats must be used appropriately to avoid bias arising from habitat-enhancing effects and/or altered predation risk. The restoration value of artificial retreats requires further testing on other species and in areas where natural retreat sites are limited.

Artificial retreats are increasingly used to sample animal populations and in attempts to boost animal numbers in degraded habitats. Here, I test potential applications of artificial retreats for lizards inhabiting a coastal environment of high conservation value (Kaitorete Spit, New Zealand). I first conducted a pitfall-trapping survey examining the distribution and relative abundance of lizards in duneland, farmland and shrubland habitats, and tested the influence of trap placement on capture rates. Capture rates of the diurnal skinks Oligosoma maccanni (McCann�s skink) and O. nigriplantare polychroma (common skink) were highest in duneland and farmland, respectively, and were most sensitive to the distance separating traps from the nearest cover (the greater the distance, the lower the capture rate). Captures of O. lineoocellatum (spotted skink) and Hoplodactylus maculatus (common gecko) were rare. Secondly, in separate chapters I test the utility of artificial retreats for monitoring: 1) a preference trial examining relative use of three types of artificial retreats by skinks (O. maccanni and O. n. polychroma) and geckos (H. maculatus); 2) a comparison of the effectiveness of artificial retreats relative to pitfall traps for detecting cryptic and primarily nocturnal geckos (H. maculatus) following translocation; and 3) capture-recapture estimation of population parameters (survival and abundance) of H. maculatus. I found that: 1) geckos strongly preferred retreats made of Onduline over corrugated iron and concrete tiles, whereas skinks exhibited no apparent preferences; 2) artificial retreats were more effective than pitfall traps for detecting geckos following translocation; and 3) monthly survival and recapture probabilities of geckos varied with age-class and over time. Estimated survival was unexpectedly low, possibly due to excessive trap spacing. I developed a new capture-recapture model specifically for population size estimation with data from artificial retreats, which gave estimates that were up to 50% greater than those predicted by conventional capture-recapture models. I caution that permanent placement of artificial retreats in long-term studies may be inappropriate for estimation of population parameters due to potential habitat-enhancement effects and/or altered predation risk. Thirdly, I conducted a capture-recapture field experiment, using a replicated Before-After-Control-impact (BACI) design, to test the relative effects of habitat manipulation (artificial retreat addition) and partial predator removal (by fencing) on annual survival of duneland skink (O. maccanni) populations. Survival increased at sites with predator exclosures, but not at control sites or following the addition of artificial retreats, either alone or in combination with a predator exclosure. The magnitude of the increase in survival for the exclosure-only treatment was small, but sufficient to change the trajectory of an apparently stable population into an increasing one, suggesting that the population is limited by predators. Predator control, but not the addition of artificial retreats, is predicted to benefit O. maccanni. To conclude, the Onduline design developed here appears to be particularly useful for sampling cryptic, terrestrial geckos; however, artificial retreats must be used appropriately to avoid bias arising from habitat-enhancing effects and/or altered predation risk. The restoration value of artificial retreats requires further testing on other species and in areas where natural retreat sites are limited.