MIGRATION PATTERNS AND ALGAL ASSOCIATIONS OF THE SPOTTED SALAMANDER, AMBYSTOMA MACULATUM, INHABITING PENN STATE ERIE WETLANDS

Paul J. Crawford, Ashley Kingston, Ashley Frank, and Nicholas Lowrie (Carley Gwin)
ABSTRACT
The Spotted salamander, Ambystoma maculatum, is one of the many organisms that inhabit the campus of Penn State Erie, The Behrend College. Habitat degradation as a result of development and construction may have negative effects on many of these organisms; the extirpation of one species may have disastrous effects on numerous other organisms. Pit fall traps were set at pond 4 located next to the athletic fields and the recently constructed BayfrontConnector. As the salamanders migrate to their natal ponds to breed, this study will determine if spatial migration patterns have been affected. The salamanders were massed, sexed, and measured, and these data were recorded, as well as the temperature and amount of precipitation each day. Migration patterns were analyzed; the overall pattern differed between 1999 and 2009. Also, algae found in the gelatinous matrix of salamander embryos will be identified both by dichotomous key and by genetic sequence information. Algae cells will be cultured in vitro, from which DNA will be isolated. Using PCR and sequencing, genes from the chloroplast will be isolated and characterized. Eventually symbiotic relationships between the algae and the embryos will be explored.

MATERIALS AND METHODS
Drift fences and pitfall traps (Fig. 3) were positioned in the same area as those from the study performed by Silver, Campbell, and Cooper (1999) before major expansion and construction began. The only pond from which captures were obtained was pond 4 (Silver et. al, 1999). The drift fences were placed around the perimeter; each morning during the breeding season the number of salamanders captured at each fence and each salamanders’ length were recorded (Fig. 4), along with the temperature and precipitation. Comparisons between differences in the number of salamanders captured this year and in the previous study will offer information regarding any possible effects the construction and development had on A. maculatum migration patterns. A dichotomous key was used to perform a preliminary identification of the algae found within the egg mass samples, which were collected during spring 2008 and stored at -80°C. TAP medium, and then nutrient agar with ampicillin, plates were used to culture the algal samples. Currently attempts are being made to culture the algae in a liquid minimal media.

TABLE I: Per Fence Contribution to Total Salamander Captures

Fence

1999 Number Captured 5 29 144 50 42 35

2009 % Number Contribution Captured 1.64 9.51 47.21 16.39 13.77 11.48 5 11 72 29 19 37

% Contribution 2.89 6.36 41.62 16.76 10.98 21.39

F9

INTRODUCTION
The Spotted salamander, Ambystoma maculatum, is found in eastern North America. Their range extends west from Nova Scotia and the Gaspé Peninsula to the northern shore of Lake Superior and south to eastern Texas and southern Georgia (Petranka, 1998). A. maculatum are most often found in deciduous forests with large amounts of water and can also be found in upland mixed or coniferous forests (Petranka, 1998). Numerous studies suggest that A. maculatum return to their natal ponds to breed, which takes place at night during early spring when temperatures are above 0ºC and there is precipitation (Silver et. al, 1999). In the previous study, the majority of A. maculatum were found in wetlands near mature forests. Pond 4 had 976 salamander captures over a four year study and fence 11 accounted for the greatest number, 597 salamanders (Silver et. al, 1999). Penn State Erie, The Behrend College has a population of A. maculatum that may have been disrupted by campus expansion and development, as well as the construction of the Bayfront Connector (Fig. 1). The increased use of athletic fields and completion of the Bayfront Connector have eliminated some of the ponds and possibly disrupted the migration of A. maculatum. We will address if the migration patterns of the Spotted salamander have changed in response to campus development and the construction of the Bayfront Connector. A. maculatum egg masses can form a symbiotic relationship with a species of algae (Chlamydomonas) embedded in the egg’s gelatinous matrix (Fig. 2); DNA from the algae in the egg masses will be isolated and chloroplast genes will be analyzed and characterized.

F 10 F 11 UF 11 F 12
Figure 3 depicts a typical drift fence with a pit fall trap. Figure 4 illustrates the method for measuring a salamander.

UF 12

RESULTS
The timing of migration and the number of migrants were significantly influenced by weather conditions (Fig. 5). Migration began on March 18 with the capture of one Spotted salamander, but the first large number of migrants was observed on March 19. Migrant numbers at each fence were compared (Table I). Overall there was a significant change in migration patterns in 2009 vs. 1999 (G-test; G=178.7, p<0.05, df=5); Upland Fence 12 showed the largest change in proportion of salamanders captured (Fig. 6). To look at fences independently, data from 1996 to 2006 were pooled with the 2009 data. Graphing z-scores vs. time, the year to year variability is accounted for, and the fence with the most apparent change in migration is Fence 10 (Fig.7). Algae samples were initially identified as belonging to the genus Chlamydomonas. Attempts to culture the algae on TAP medium and ampicillin plates were unsuccessful.

FUTURE DIRECTIONS
The development of athletic fields and construction of the Bayfront Connector have had effects on the migration patterns of the Spotted salamander population at Penn State Erie. Future studies may expand to collect data from all the wetlands used in the original study and analyze migration patterns campus-wide. After the algae are successfully cultured, DNA will be isolated and Chlamydomonasspecific primers will be used to confirm the particular algal species associated with the egg masses, and to isolate and characterize chloroplast genes.

BIBLIOGRAPHY
Petranka, J. 1998. Salamanders of the United States and Canada. Washington and London: Smithsonian Institution Press. Silver et. al. 1999. The Population Biology of the Spotted Salamander, Ambystoma Maculatum , Inhabiting Penn State Erie Wetlands. Wyman, R.L. “What’s happening to amphibians?” Conservation Biology 4:350-352 (1990).

ACKNOWLEDGEMENTS
A special thanks to Dr. Robert Light and the Behrend Undergraduate Research Grants, Dr. Pamela Silver, and Carley Gwin. printed by
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Figure 1 depicts the location of the wetlands within the athletic fields and bordering the Bayfront Connector. Figure 2 shows an egg mass of A. maculatum and the associated algae.

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