Intraspecific phylogeography of the gopher tortoise, Gopherus polyphemus: RFLP analysis of amplified mtDNA segments

The slow rate of mtDNA evolution in turtles poses a limitation on the levels of intraspecific variation detectable by conventional restriction fragment surveys. We examined mtDNA variation in the gopher tortoise (Gopherus polyphemus) using an alternative restriction assay, one in which PCR-amplified segments of the mitochondrial genome were digested with tetranucleotide-site endonucleases. Restriction fragment polymorphisms representing four amplified regions were analysed to evaluate population genetic structure among 112 tortoises throughout the species' range. Thirty-six haplotypes were identified, and three major geographical assemblages (Eastern, Western, and Mid-Florida) were resolved by UPGMA and parsimony analyses. Eastern and Western assemblages abut near the Apalachicola drainage, whereas the Mid-Florida assemblage appears restricted to the Brooksville Ridge. The Eastern/Western assemblage boundary is remarkably congruent with phylogeographic profiles for eight additional species from the southeastern U.S., representing both freshwater and terrestrial realms.     

Salmonella from gopher tortoises (Gopherus polyphemus) in south Georgia

From 2002 to 2006, gopher tortoises (Gopherus polyphemus) were collected at Moody Air Force Base, Lowndes/Lanier counties, Georgia, USA, and opportunistically surveyed for the presence of Salmonella species. Four of 155 (2.6%) cloacal swabs collected from 80 tortoises were positive for the presence of Salmonella enterica, and the following serovars were identified: Give, Hartford, Javiana, and Luciana. Female tortoises (5%) were infected at a rate similar to male tortoises (5%). All isolates were obtained from adult tortoises (n = 73); subadults (n = 7) were all negative. Each isolated serovar is a potential human pathogen, suggesting appropriate precautions should be emphasized when handling these animals.     

Morphological changes in the oviductal endometrium during the reproductive cycle of the tortoise,Gopherus polyphemus

The oviducts of 24 tortoises (Gopherus polyphemus) were examined using histological techniques and scanning electron microscopy to determine endometrial morphology. Measurements of endometrial characteristics (epithelial cell height, cilia length, thickness of endometrial glandular layer, and glandular diameter) in the uterus and tube (tuba uterina) were obtained to determine changes during the reproductive cycle. Epithelial cell height increases in both the uterus and the tube during vitellogenesis and remains hypertrophied during gravidity. Cilia length increases in the uterus during late vitellogenesis and gravidity, but the length of tubal cilia does not change during the reproductive cycle. The ratio of secretory to ciliated epithelial cells in the oviduct increases from quiescence to gravidity. The thickness of the glandular endometrial layer increases in both the uterus and tube during vitellogenesis. In the uterus, the glandular layer decreases in thickness during gravidity. The diameter of the uterine glands increases throughout vitellogenesis and gravidity; however, following ovulation glandular cells become depleted of secretory granules and cell height diminishes. The diameter of the tubal glands is unchanged during the reproductive cycle. Oviductal hypertrophy during vitellogenesis coincides with elevated circulating estradiol, whereas during gravidity progesterone concentrations peak (Taylor, '82, PhD Dissertation, University of Florida, Gainesville) and may induce secretion of albumen and eggshell components.     

Histology and functional morphology of the female reproductive tract of the tortoise Gopherus polyphemus

The oviducts of 25 tortoises (Gopherus polyphemus) were examined by using histology and scanning electron microscopy to determine oviductal functional morphology. Oviductal formation of albumen and eggshell was of particular interest. The oviduct is composed of 5 morphologically distinct regions; infundibulum, uterine tube, isthmus, uterus, and vagina. The epithelium consists of ciliated cells and microvillous secretory cells throughout the oviduct, whereas bleb secretory cells are unique to the infundibulum. The epithelium and endometrial glands of the uterine tube histologically resemble those of the avian magnum which produce egg albumen and may be functionally homologous. The isthmus is a short, nonglandular region of the oviduct and appears to contribute little to either albumen or eggshell formation. The uterus retains the eggs until oviposition and may form both the fibrous and calcareous eggshell. The endometrial glands are histologically similar to the endometrial glands of the isthmus of birds, which are known to secrete the fibers of the eggshell. These glands hypertrophy during vitellogenesis but become depleted during gravidity. The uterine epithelium may supply "plumping water" to the egg albumen as well as transport calcium ions for eggshell formation. The vagina is extremely muscular and serves as a sphincter to retain the eggs until oviposition. Sperm are found within the oviductal lumen and endometrial glands from the posterior tube to the anterior uterus throughout the reproductive cycle. This indicates sperm storage within the female tract, although the viability and reproductive significance of these sperm are unknown.     

Orientation of the gopher tortoise, Gopherus polyphemus

Twenty-three gopher tortoises were released in an open field at least 8 km from their home burrows, and thirty-seven were tested in an arena which excluded all directional landmarks. Over half showed a statistically significant individual direction preference under each of these conditions. Directions chosen had no relation to the homeward direction and were widely divergent for individuals from the same geographic population. Comparing the preferred directions in the open field and in the arena indicates that landmarks may have a disruptive effect on orientation. The preferred headings of ten tortoises subjected to a 6-hr phase shift was about ninety degrees in the expected direction, indicating that this species is capable of sun-compass orientation.     

Polymorphic microsatellite markers for the Mojave desert tortoise, Gopherus agassizii

We describe primers and polymerase chain reaction (PCR) conditions to amplify 14 tri- and tetranucleotide microsatellite loci for the Mojave desert tortoise (Gopherus agassizii). Across three populations (87 individuals) located in the Mojave Desert, USA, the markers yielded a range of four to 33 alleles and an average observed heterozygosity of 0.733 (range 0.433 to 0.933). We neither detected linkage disequilibrium between any pair of loci nor did we find a consistent pattern of deviation from Hardy-Weinberg equilibrium. These microsatellites are designed for PCR multiplexing, and provide higher throughput capacity to aid in conservation genetics studies for this threatened species.     

Population and conservation genetics of the gopher tortoise (<Emphasis Type="Italic">Gopherus polyphemus</Emphasis>)

The gopher tortoise (Gopherus polyphemus) is an important member of the sandhill, longleaf pine, and scrub ecosystems in the southeastern United States. Even though it is currently protected throughout its range, tortoise populations continue to decline. We assessed genetic diversity at nine microsatellite loci in 300 individuals from 21 locations throughout Florida and southern Georgia. Tortoise populations are clearly subdivided into at least eight genetic assemblages with an . Furthermore, we found indications of anthropogenic effects in the form of population bottlenecks in five populations and putative admixture in four. From these data, we recommend that the populations be managed to maintain existing genetic structure without further isolation of populations and the establishment of a holistic␣database to include genetic and demographic information useful for relocation and management purposes.     

Histomorphometric studies of dermal bone in the desert tortoise, Gopherus agassizii.

Dermal bone biopsies were collected from the periphery of the carapaces of adult desert tortoises (Gopherus agassizii) from grazed and ungrazed habitats near the Arizona/Utah border (USA). Quantitative bone histomorphometry was performed on these biopsies as well as on dermal bone biopsies collected from captive juvenile desert tortoises. Except for mild osteomalacia, carapaces of adult desert tortoises from the grazed habitat were relatively normal. No signs of osteopenia were observed. Based on the low numbers of osteoblasts and osteoclasts in dermal bone of both populations of adult desert tortoises, it appears that the peripheral carapace is relatively inert with very low levels of dermal bone turnover. Bone cells and osteoid were more common in dermal bone biopsies from the carapace and plastron of captive juvenile desert tortoises than in adult desert tortoises. However, the great variability in the incidence of bone cells among individuals and the difficulty in collecting juvenile desert tortoises in the field limit the usefulness of dermal bone biopsies from animals of this age group. Based on these results, we propose that dermal bone of the peripheral carapace is a poor sample site for evaluating the effects of dietary or environmental conditions on calcified tissues in desert tortoises.     

PCR primers for microsatellite loci in the desert tortoise (Gopherus agassizii,Testudinidae)

The desert tortoise, Gopherus agassizii, is a threatened species native to the North American desert southwest and is recognized as having distinct Mojave and Sonoran populations. We identified six polymorphic microsatellite loci in the desert tortoise. All six loci were polymorphic in Sonoran samples. Five of the loci were variable in Mojave samples with varying degrees of amplification success. Two of the loci exhibited low allelic variation (2–3 alleles) while four were highly variable (8–27 alleles).     

Effects of predator exclusion on nest and hatchling survival in the gopher tortoise

Gopher tortoise (Gopherus polyphemus) populations are declining throughout the Southeast, and high levels of predation on nests and juveniles have been suggested as a potential contributor to this decline. Therefore, we documented gopher tortoise nest success and hatchling survival relative to mammalian predator control. We used 4, large (approx. 40-ha) fenced, predator exclosures to exclude mid-sized mammalian predators: bobcat (Lynx rufus), raccoon (Procyon lotor), Virginia opossum (Didelphis virginianus), fox (Urocyon cinereoargenteus and Vulpes vulpes), coyote (Canis latrans), nine-banded armadillo (Dasypus novemcinctus), and skunk (Mephitis mephitis); 4 unfenced plots served as controls. We monitored nests for survival through hatching and used radio-telemetry to examine hatchling survival. We radio-tracked 40 hatchlings for up to 329 days, but we were only able to track 8 individuals from a single nest at an unfenced plot because of high nest predation. Mean nest survival was greater at exclosures than at unfenced controls (F_{1, 2} = 45.80, P = 0.0001). Hatchling survival differed (χ² = 5.839, P = 0.016) between unfenced plots (37.5%) and exclosures (74.4%), suggesting that mammals also were significant predators of hatchlings. The number of juvenile (<13 cm in diameter) and subadult tortoise burrows (13–21.9 cm) increased over a 6-year period in exclosures, providing further support for an effect of excluding mammalian predators on nest and juvenile tortoise survival. © 2012 The Wildlife Society.     

Extrapair paternity, migration, and breeding synchrony in birds

To understand interspecific patterns in the strength of sexualselection, variation in the costs and benefits of exercisingmate choice needs to be evaluated. One manifestation of sexualselection in birds is the occurrence of greatly variable levelsof extrapair paternity (EPP). A proposed general explanationfor this variation is that the benefits to females in seekingextrapair copulations vary in a predictable manner accordingto the degree of breeding synchrony because females are betterable to assess potential extrapair partners when males are simultaneouslyin breeding condition. This hypothesis predicts a latitudinaltrend in EPP because birds tend to breed more synchronouslyaway from the equator. Expanding on previous geographicallyand taxonomically restricted tropical/temperate comparisons,we used phylogenetically independent standardized linear contraststo show that this positive relationship persists when all birdspecies for which EPP estimates currently exist are considered.However, if a third factor covaries with latitude in the sameway as breeding synchrony and EPP, this relationship need notbe causal. Migration could also account for latitudinal variationin the benefits to females of pursuing EPP, if migration isassociated with (1) hasty or (2) inaccurate mate choice, (3)facilitated assessment of male quality through the condition-dependenceof arrival time, or (4) increased genetic variance in male quality.We show that migration distance is positively related to theproportion of EPP and that migration can statistically explainthe latitudinal trend in EPP, even when confounding factorsare simultaneously controlled. Hence, alternative explanationsfor latitudinal variation in EPP may be feasible, and carefulintraspecific tests are needed to assess their relative importanceand their implications for geographical variation in life-historyevolution.     

Group structure, mating system and extra-group paternity in the co-operatively breeding White-breasted Thrasher Ramphocinclus brachyurus

Avian co-operative breeders show a diverse range of social and reproductive systems. Here we combined microsatellite genotyping with field observations over three consecutive breeding seasons to investigate the social structure and mating system of the White-breasted Thrasher Ramphocinclus brachyurus . Co-operative breeding was facultative in this species, with approximately one-third of nests having helpers. Breeding groups comprised a pair of breeding adults and up to three helpers of either sex. Within co-operative groups, age was associated with parentage in both sexes. Helpers were retained offspring and either rarely or never achieved parentage in current broods. There was no evidence of egg dumping and a relatively small percentage of chicks (7.5%, n  = 67) were fathered by males from outside the co-operative group. The White-breasted Thrasher is a globally Endangered species with a very small and declining population. A captive breeding programme for this species is not recommended at present, but may become necessary in the future, and any such programme should take account of our findings regarding breeding behaviour and group structure.     

Distribution and habitat utilization of the gopher tortoise tick (Amblyomma tuberculatum) in Southern Mississippi

The distribution of the gopher tortoise tick ( Amblyomma tuberculatum ) has been considered intrinsically linked to the distribution of its primary host, gopher tortoises ( Gopherus polyphemus ). However, the presence of G. polyphemus does not always equate to the presence of A. tuberculatum . There is a paucity of data on the ecology, habitat preferences, and distribution of A. tuberculatum . The goals of this study were to assess the distribution of A. tuberculatum in southern Mississippi and to determine which, if any, habitat parameters explain the distribution pattern of A. tuberculatum . During 2006-2007, we examined 13 G. polyphemus populations in southern Mississippi for the presence of A. tuberculatum , and we measured a suite of habitat parameters at each site. Only 23% of the G. polyphemus populations supported A. tuberculatum , suggesting a more restricted distribution than its host. The results of our multivariate analyses identified several habitat variables, e.g., depth of sand and percentage of sand in the topsoil and burrow apron, as being important in discriminating between sites with, and without, A. tuberculatum. Amblyomma tuberculatum was only found at sites with a mean sand depth of > 100 cm and a mean percentage of topsoil and burrow apron sand composition > 94.0 and 92.4, respectively. Thus, environmental factors, and not just its host's range, seem to influence the distribution of A. tuberculatum.     

Multiple paternity in the freshwater snail,Potamopyrgus antipodarum

Mating multiply may incur costs, such as exposure to predators and to sexually transmitted diseases. Nevertheless, it may be favored, in spite of these costs, as a way to increase the genetic diversity of offspring through fertilization by multiple males. Here, we tested for multiple paternity in a freshwater snail (Potamopyrgus antipodarum), which is host to several species of sterilizing trematode worms. Using microsatellites markers, we found multiple paternity in two different snail populations, with as many as seven males fertilizing a single female. In addition, high evenness of sire fertilization was found within individual broods. Multiple paternity can occur for a variety of reasons; however, given that these populations experience high risk of infection by a sterilizing trematode, one potential explanation may be that multiple paternity and high evenness of sire fertilizations increase the chances of the production of parasite‐resistant offspring.