ASB 2004 - Memphis TN





Marshall University and Fernbank Science Center


Sexual dimorphism in the eastern hellbender, Cryptobranchus a. alleganiensis


The Eastern Hellbender, Cryptobranchus a. alleganiensis, is a large, aquatic salamander found in higher order, cold-water streams in the Ohio River drainage. Males will commonly fight for possession of the best breeding sites (usually under rocks) and will also guard the eggs they fertilize. Past research has shown that when urodelan species practice male combat, the males normally reach a size as large if not larger than females. Based on these behaviors, it would be expected that males become larger than females. Contrary to this prediction, it is thought that females reach a larger overall length than males. Due to tremendous size overlap though, determining gender based simply on total length (TL) can be nearly impossible. This research examined whether there are other sexual dimorphic characters present all year. Preserved specimens (n=105, f=52, m=53) from different populations were measured for TL, snout-vent length (SVL), thoracic girth (TG), head width (HW), and mass. Gender was determined by checking for the presence of follicles. All measurements were then divided by the specimenís TL or SVL, so size-corrected ratios were also compared. Pair-wise comparisons (t-tests) were made between males and females for each measurement and ratio. Males were found to differ significantly from females for the ratio TG/TL (P<.05.) Principal component analyses showed no separation between males and females, but logistic regression did reveal that females are more variable than males. Based on these results, the dimorphism that exists in hellbenders is not great enough to positively determine the gender of specimens.




Department of Biological Sciences, Marshall University, Huntington, West Virginia


Phenotypic variation among cave-dwelling spring salamanders, Gyrinophilus spp. Cope (Plethodontidae), in West Virginia, Virginia, and Tennessee


Green and Brantís 1966 survey of salamanders in West Virginia caves revealed that Spring Salamanders, Gyrinophilus spp., are the most frequently encountered salamanders in subterranean habitats in the state. In 1977, Bersharse and Holsinger described Gyrinophilus

subterraneus, the West Virginia Spring Salamander, from one cave population in Greenbrier County. Some have suggested that G. subterraneus is an extreme variant of the widespread G. porphyriticus. In an attempt to document the degree of variability among cave-dwelling Gyrinophilus species, Principle Components Analysis (PCA) was applied to measurements of 20 external morphological characters. This analysis was based on 106 specimens of cavedwelling Gyrinophilus spp. from the U. S. Natural History Museum (USNM) and West Virginia Biological Survey collection (WVBS). Specimens analyzed included; 5 metamorphosed and 11 larval G. subterraneus, 33 metamorphosed and 24 larval G. porphyriticus, 1 metamorphosed and 20 neotenic G. palleucus palleucus, and 6 neotenic G. gulolineatus. Eye diameter was the source of the greatest variation between species. Separation was pronounced between G. p. palleucus and G. gulolineatus and weaker between G. subterraneus and G. porphyriticus, where some overlap occurs in PCA. A grant from the West Virginia Division of Natural Resources Wildlife Diversity Program supported this research.




Department of Biological Sciences, Marshall University, Huntington, West Virginia


The natural history of cavedwelling spring salamanders, Gyrinophilus spp. Cope (Plethodontidae), in West Virginia.


Surface populations of Spring Salamanders, Gyrinophilus porphyriticus ssp., have been widely studied in cool mountain streams, seeps, and springs throughout their broad range in the Appalachian region. In West Virginia, however, they are also commonly found in caves. Green and Brantís 1966 survey of salamanders in West Virginia caves revealed that G. porphyriticus are the most abundant salamanders in subterranean habitats in the state. In 1977, Besharse and Holsinger described the West Virginia Spring Salamander, Gyrinophilus subterraneus, from one cave population in Greenbrier County. Since then, there has been little investigation into the natural history of cave-dwelling Spring Salamanders. We monitored two cave populations of G. porphyriticus using mark-recapture techniques with the objective of investigating the ecology, life histories, and population structure of these troglophilic salamanders. This study has provided insight into the secretive lives of some of the top predators of West Virginia cave ecosystems and should be valuable for the conservation of these species. Grants from the West Virginia Division of Natural Resources Wildlife Diversity Program and the West Virginia Association for Cave Studies supported this research.




Department of Biology, Marshall University and West Virginia Division of

Environmental Protection


The use of streamside salamanders as indicators of headwater stream health in West Virginia


The health of headwater streams in the United States has been assessed using various types of bioindicators, including benthic invertebrates and fishes. In our study funded by the USGS and the US EPA we examined the potential use of streamside salamanders as another type of bioindicator. Forty-four randomly selected sites in West Virginia were surveyed using complete census and multiple pass removal sampling techniques over the course of 2 years.  Environmental parameters were recorded and included temperature, pH, relative humidity, turbidity, cobble count, etc. Using the program CAPTURE, we were able to calculate detection probabilities and population estimations from our removal sampling. Multivariate regression analysis was also used and showed correlations between habitat data and salamander abundance. This data indicates that stream salamanders can serve as bioindicators to headwater stream health.




Alabama A&M University and USDA Forest Service, Southern Research Station


Demographics of Bufo americanus populations in relation to several silvicultural techniques in northern Alabama


The eastern American toad is usually considered a habitat generalist. However, we predicted that demographics of toad populations would vary in relation to different levels of overstory tree retention. We predicted that, due to physiological constraints, smaller sized individual would be encountered less often on clearcuts and other treatments with low levels of basal area. Sex ratios might also differ because of differing survival rates. There seemed to be a relationship between body size (SVL and mass) and treatment, with larger average body sizes on clearcut and control plots, and smaller sizes in plots with intermediate levels of harvest. Sex ratios were not significantly different among treatments. The ratio of juvenile to adult toads also seemed to vary by treatment. These results suggest that subtle changes in population structure of American toads may occur in response to habitat changes associated with silviculture.




University of Memphis


Effect of hydroperiod on developmental polymorphisms of the eastern newt


Recent studies provide evidence that ecological factors play a role in parapatric speciation.

There are four subspecies of the eastern newt (Notophthalmus viridescens) distributed parapatrically and this species expresses developmental polymorphisms, which reflect adaptations to variable environments. Larvae potentially can become terrestrial efts, aquatic lunged adults, or aquatic gilled adults. Pond hydroperiod is an important environmental parameter for aquatic amphibians, and it varies regionally. Previous work indicates that frequency of developmental polymorphisms varies among subspecies. Our research tested the hypothesis that there were genetically based differences among subspecies in developmental polymorphisms in relation to pond hydroperiods. We collected adults of three subspecies (N. v. viridescens, N. v. dorsalis, and N. v. louisianensis) for breeding. Hatched larvae were distributed into 36 replicated artificial ponds with different hydroperiods: constant water, short hydroperiod (4 week-drying), and long hydroperiod (8 week-drying). We measured four responses: survival, mass at metamorphosis, larval period, and developmental outcome. Our result indicated that N. v. viridescens produced a significantly higher proportion of efts than N. v. dorsalis under shorter hydroperiods. Under longer hydroperiods, N. v. viridescens had shorter larval periods than N. v. dorsalis, but had significantly smaller metamorphic body masses than both N. v. dorsalis and N. v. louisianensis. These results suggest N. v. viridescens may be more adapted to shorter hydroperiods, whereas under longer hydroperiods, the other two subspecies perform better. Accordingly, strong selection imposed by different hydroperiods could lead to life history divergence and potentially speciation if accompanied by assortative mating within subspecies.




Marshall University


Reproductive biology of Regina septemvittata (Queen Snake) in West Virginia


Reproductive biology of Regina septemvittata was studied in Ohio County, West Virginia during the 2003 summer season. Ten gravid snakes were collected between 28 July and 7 August and housed in aquaria until parturition. Data collected on adult female snakes included snout-vent length, total body length, pre-parturition mass, post- parturition mass, relative clutch mass, and behavior during capture. Data collected on neonates included snout-vent length, total body length, mass, and gender. Number of neonates per clutch ranged between 6 and 10 with a mean number of 8 per clutch. Of 81 neonates, 36 were male and 45 were female. The mean relative clutch mass for adult females was 0.34. Gravid females were also notably more aggressive than non-gravid snakes during capture. When West Virginia R. septemvittata were compared to a Kentucky population several differences were observed. Adult Kentucky snakes had a significantly higher snout-vent lengths (P = < .001) and mean clutch size (P = < .001) than West Virginia R. septemvittata. However preparturition mass did not differ significantly (P = .264). Between the two localities, relative clutch mass was not significantly different. Neonate West Virginia R. septemvittata had significantly smaller snout-vent lengths (P < .001) and masses (P<.001). Overall West Virginia R. septemvittata were smaller than Kentucky snakes in all but one reproductive aspect (preparturition mass). These findings concur with the hypothesis that as latitude increases, size decreases within a snake species.




Department of Biological Sciences, Marshall University, Huntington, WV.


Discovery of Aeromonas hydrophila and Pseudomonas spp. skin infections and other malformations of Rana pipiens in West Virginia


Over the past few decades, biologists have been attempting to uncover the cause(s) of the Global Amphibian Decline (GAD). There are many potential factors linked to the GAD, such as increased UV light, habitat degradation, and pollution. More recently, researchers are discovering that infections and malformations are also major components of the GAD. In 2003, two male specimens of Rana pipiens were confirmed to have skin infections caused by Red-leg disease, Aeromonas hydrophila, at Greenbottom Swamp in Cabell County, WV.  Using sterile techniques, two distinct bacterial colonies were isolated from ulcers on the frogsí legs. Using Biolog, the colonies were positively identified as Aeromonas hydrophila (99.2% +/- 0.374) and mixed colonies composed of Pseudomonas fulva and P. maculicola. Three additional specimens of Rana pipiens and one of R. catesbeiana were discovered with hindlimb malformations, including undeveloped phalanges, malformed toes, and polydactyly. Also, numerous specimens of R. pipiens (n= 31) were found with skin tumors on the urostyle, rostrum, lip, and dorsal surface. The presence of infections and tumors suggest that the immune system of the affected frogs is compromised by some additional stress, such as the factors listed above. The presence of malformations and Aeromonas hydrophila infections in these frog populations should serve as an indicator of a greater problem occurring at this particular swamp. Funding: WVDNR.







Department of Biological Sciences, Marshall University, Huntington, West Virginia


Use of an artificial pond by amphibians and reptiles in West Virginia


A farm pond approximately 40 years old and located in Wayne County, West Virginia was studied to determine the ingress and egress of amphibians and reptiles. The period of study extended from February 2003 to November 2003. The pond is approximately 50 m long and 25 m wide and is located on a south-facing hillside at 622 ft. in elevation. A drift fence composed of landscaping cloth was constructed to completely encircle the site. Funnel traps were positioned on both sides of the fence every 5.5 meters and all traps were checked daily during the study period. The most common species found entering and exiting the pond were (in order of frequency) Rana clamitans melanota, Notophthalmus v. viridescens, Pseudacris c. crucifer, and Bufo a. americanus. Other species less frequently observed included R. palustris, R. catesbeiana, P. brachyphona, Ambystoma maculatum, Terrapene c. carolina, and Scincella lateralis. Additionally, ten other species were trapped while entering or leaving the pond. This is the first extended study of the use of an artificial pond in West Virginia and demonstrates that farm ponds can provide habitat for reproduction and foraging for many amphibian and reptile species. With the rapid loss of natural pools and ponds due to habitat alterations, artificial aquatic systems play an important role in the preservation of amphibians and reptiles in the central Appalachian Mountains.




Department of Biological Sciences, Marshall University, Huntington, WV


Analysis of anuran community level interactions at Greenbottom Swamp in Cabell County, WV


Community level studies are very important in determining the extent of niche partitioning between different frog species. In this study, community level interactions between Rana pipiens, R. clamitans melanota, and R. catesbeiana were analyzed from September 2003 until November 2003. These frog species were chosen because there appears to be exploitation of different niches. In this study, stratification along terrestrial and aquatic niches was measured in a marsh at Greenbottom Swamp. Frogs were captured within a 60m x 32m quadrant and their positions were marked with flags. The distance from the water/land interface was measured and relative positions were analyzed. There was significant niche separation between R. catesbeiana and R. pipiens (P= 0.001). There was also niche separation between R. clamitans melanota and R. catesbeiana and between R. clamitans melanota and R. pipiens, however these distances were not significant. The results were as follows: R. pipiens was the most terrestrial, and R. catesbeiana was the most aquatic, while R. clamitans melanota was found in both terrestrial and aquatic habitats. Additionally, within the species groups, there was separation into different size classes. Rana pipiens separated into two size classes, while R. catesbeiana separated into three. From these results, it is apparent that there is niche partitioning between the above listed frog species and that different size classes exist among these species. Funding: WVDNR.




Shorter College


Diet of Eurycea cirrigera larvae in a woodland stream


We examined the stomach contents of 48 Eurycea cirregera larvae in a fishless spring-fed stream (eight larvae were sampled every 4 h over a 24 h period). In addition, we characterized the benthic macroinvertebrate fauna in the creek to determine the available prey base of E. cirregera larvae. Overall, E. cirregera larvae captured more prey during the day than night and total prey abundance was greatest at 0800. The number of prey decreased throughout the remaining day and night collections and reached its lowest abundance at 0400. The diet of E. cirregera larvae consisted mostly of isopods, amphipods, ephemeropterans, trichopterans and chironomids. These taxa were commonly found in our macroinvertebrate sample as well, but their proportion in the diet in some cases suggested preference or avoidance of certain prey taxa. These gape limited salamanders showed selection for smaller prey (chironomids and amphipods) and some avoidance of larger prey such and isopods and trichopterans, whose size and/or behavior (case building by trichopterans) may deter or minimize predation.




Department of Biology, Marshall University


Effectiveness of turtle trapping techniques in West Virginia


Many methods of inventory have been used worldwide to capture and observe turtle species.  Some techniques include live capture with traps and nets and visual surveys with binoculars and spotting scopes. During the summer of 2003, a statewide inventory of West Virginia riverine turtles was implemented employing some of these methods. Live capture was conducted with fyke nets, catfish traps, basking traps, and large hoop nets. Visual surveys were done with spotting scopes and binoculars. The data was analyzed to determine capture efficiency and effectiveness for each method and includes a discussion on the advantages and disadvantages for each technique.




Marshall University


Amphibian and reptile surveys in the Gauley River National Recreation area in West Virginia


The Gauley River National Recreation area (GRNR) is located in central West Virginia in the Allegheny Plateau region. The study area is located downstream of Summersville Lake and consisted of the Gauley River, its major tributaries, and the associated forested/riparian habitats. From 2000 to 2003, searches were conducted at all times of the year with a variety of sampling techniques. Collectively, 45 species of reptiles and amphibians were documented.  Of these, 8 are considered species of special concern. Certain habitats were found to possess much greater herpetofauna diversity than others. Some species that were expected to be there (i.e. Eumeces a. anthracinus) were not found, but this is probably due more to the difficulty in capturing these species than their absence. Future surveys will focus on locating the expected species that have not yet been found.




Marshall University and State University of New York, Syracuse


Status of the West Virginia state collection of amphibians and reptiles


The West Virginia Academy of Science gave Neil D. Richmond $100 in 1935 to travel the state and collect amphibians and reptiles. These specimens and supplemental collections in 1937 and 1938 formed the nucleus for a state collection of herpetofauna. Since Richmond was not associated with a museum or University, he lacked curatorial services and a building to hold the collections. To provide curatorial services the collections were moved to Marshall College in 1939 under the care N. Bayard Green. N. B. Green maintained the collections from 1939 Ė 1971. During this time, the collections grew from approximately 1,000 to over 5,000. Michael Seidel served as curator from 1971 to 1987. Thomas K. Pauley assumed the curatorship in 1987 and continues to provide curatorial services for the collection today. Presently there are over 14,000 specimens. The WV Division of Natural Resources, United States Park Service, and United States Department of Agriculture- Forest Service have provided financial service for the maintenance of the collection.