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In this project, red-backed salamanders, Plethodon cinereus, were collected from the heart of their range (Mountain Lake, Virginia, USA), in an effort to document the typical, or baseline, leukocyte profile of this species in its natural state. From microscopic examination of thin blood films, we determined relative proportions of all leukocyte types and determined the ratio of neutrophils to lymphocytes, which is a useful index of stress levels. In addition, individuals were examined for three naturally occurring endoparasites: two gastro-intestinal (trematodes and a ciliated protozoan in the genus Cepedietta) and one in the blood (a Rickettsial bacteria). This allowed us to conduct statistical comparisons of leukocyte data across parasite infection groups. Of 36 salamanders, 11 (31%) had trematode parasites, 7 (19%) contained Cepedietta parasites, 6 (17%) were parasitized by Rickettsia, and 12 had no parasites. The most common leukocytes were lymphocytes (87.4%), followed by neutrophils (4.2%), monocytes (3.3%), eosinophils (2.6%), and basophils (2.5%). The average neutrophil/lymphocyte ratio for these salamanders was remarkably low (0.05) for an amphibian population. Statistical comparisons of leukocyte profiles among parasite infection groups indicated parasitized salamanders had twice as many circulating leukocytes as nonparasitized individuals, which could be considered an "inflammation response". There was a slight but significant elevation in relative eosinophil abundance in parasitized salamanders. There were few differences in leukocyte subsets between parasite types, nor was there evidence of any proliferation of phagocytic cells in any parasitized individual. [ABSTRACT FROM AUTHOR]

Parasitic leeches and trypanosomes release chemical signals into their hosts to evade immuno-detection, but it is unknown whether these compounds manipulate host behavior or endocrine physiology. We determined whether parasitic infections with leeches and/or trypanosomes affected the immune and stress response of an imperiled giant species of amphibian, the eastern hellbender (Cryptobranchus alleganiensis Daudin). We monitored corticosterone and white blood cell counts in response to restraint and injection with adrenocorticotropin hormone (ACTH) or saline for up to 50 h. The presence of leeches dampened hellbender corticosterone responses to restraint and reduced diel patterns of plasma corticosterone. Injection with ACTH restored the normal inter-renal responses of hellbenders, suggesting that leeches, possibly through neurotransmitters in leech saliva, cause down-regulation of corticosterone release at the level of the pituitary or hypothalamus. Infection with leeches also increased the relative abundance of eosinophils, white blood cells often recruited into circulation in response to parasitic infection. Lastly, neutrophil to lymphocyte (N:L) ratios increased in all animals after 24 h of capture and remained elevated for up to 50 h, but these temporal dynamics did not differ with parasite infection. Trypanosome infection did not affect any aspect of hellbender physiology that we measured. Our findings reveal a previously undocumented host-parasite dynamic. While the functional significance to the parasite is unclear, the physiological and behavioral implications for the host are great, given the important role of glucocorticoids in regulating physiology and behavior., (© 2015. Published by The Company of Biologists Ltd.)

Eastern hellbender salamanders (Cryptobranchus alleganiensis alleganiensis) are declining in North America and because of this the health status of individuals in several populations is closely monitored by researchers. During a health survey of hellbenders from a stream in Smyth County, VA, USA, we examined Giemsa-stained blood smears of 71 animals captured during 2011 for the presence of blood parasites. We discovered an unknown species of trypanosome that was apparently widespread within this population; 40 of the 71 individuals (56.3 %) were infected. There are seven known trypanosome species of caudate amphibians; based on microscopic examination, the parasite we observed appeared most similar to Trypanosoma cryptobranchi, which was reported in this species only once before, 76 years ago, from a single animal apparently captured in Iowa. Given that some trypanosomes can adversely affect the health of their hosts, we recommend further monitoring be done in this and other hellbender populations to determine the geographic extent of the parasite and its effects on its increasingly rare host.