Mouse monoclonal antibodies are preferred over rabbit polyclonal antibodies because they are secreted by immortalized hybridomas in vitro, and thus there would be no need to immunize mice or rabbits in the future. scientific community interested in disease ecology of alligators. == Abstract == Little is known about the disease ecology of American alligators (Alligator mississippiensis), and especially how they respond immunologically to emerging infectious diseases and zoonotic pathogens. In this study, we examined serum samples collected from wild alligators in Florida (20102011) and South Carolina (20112012, 20142017) for antibody responses to multiple bacteria. Immunoglobulin Y (IgY) was purified from serum to generate a mouse monoclonal antibody (mAb AMY-9) specific to the IgY heavy chain. An indirect ELISA was then developed for quantifying antibody responses against whole cellEscherichia coli,Vibrio parahaemolyticus,Vibrio vulnificus, Mycobacterium fortuitum, Erysipelothrix rhusiopthiae, andStreptococcus agalactiae. In Florida samples the primary differences in antibody levels were between JanuaryMarch and late spring through summer and early fall (May-October), most likely reflecting seasonal influences in immune responses. Of note, differences over the months in antibody responses were confined toM. fortuitum,E. rhusiopthiae,V. vulnificus, andE. coli. Robust antibody responses in SC samples were observed in 2011, 2014, and 2015 against each bacterium exceptE. coli. All antibody responses were low in 2016 and 2017. Some of the highest antibody responses were againstV. parahaemolyticus,M. fortuitum, andE. rhusiopthiae.One SC alligator estimated to be 70+ years old exhibited the highest measured antibody response againstV. parahaemolyticusandM. fortuitum.By combining data from both sites, we show a clear correlation between body-mass-indices SB 399885 HCl (BMI) and antibody titers in all six of the bacteria examined. Our study provides a critical antibody reagent and a proof-of-concept approach for studying the SB 399885 HCl disease ecology of alligators in both the wild and in Rabbit Polyclonal to WIPF1 captivity. Keywords:American alligator, IgY, mAb AMY-9, disease ecology, immune responses, environmental immunology == 1. Introduction == The American alligator,Alligator mississippiensis, is a crocodilian distributed throughout the Southeastern USA, from Texas to North Carolina [1]. Due to heavy and largely unregulated hunting pressure from the 1870s to the 1960s, these crocodilians dwindled in numbers until they were protected under the Endangered Species Conservation Act of 1969 (a precursor to the Endangered Species Act of 1973), and thereafter removed from the list in 1987 as populations recovered [2,3]. Today, although alligators are categorized as a species of Least Concern by the International Union for Conservation of Nature [4], populations remain vulnerable to multiple conservation threats including habitat degradation, pollution, invasive species, and climate change [5]. Reproductive traits of alligators are key factors in the ability of these animals to withstand environmental stressors, and healthy habitat and adaptability to changing environmental factors are essential to reproductive success [6,7,8]. However, the crocodilian immune system is also impacted by environmental stressors (e.g., pathogens), and a healthy immune system is a key measure of fitness in terms of ecological and evolutionary success [9,10,11]. Compared to other vertebrates, the link between basic immunobiology and effects of environmental stressors on immune function in reptiles is not well understood, though this field is rapidly advancing [12,13]. While most aspects of innate immunity, including biochemical and cellular components are conserved across all vertebrates, much less is known about SB 399885 HCl pathogen-specific immune responses in reptiles [14,15], SB 399885 HCl particularly crocodilians. Only a small number of studies have examined the effects of environmental contaminants on crocodilian immune function, and even fewer have assessed the immunological effects of bacterial pathogens on these animals. Thus, there is a clear need for SB 399885 HCl advancement in our understanding of immunological responses of free-ranging crocodilians to naturally occurring pathogens within their habitats. A key factor in understanding and possibly managing the pathogenic component of environmental stressors is monitoring the presence of various pathogens, and especially those deemed to be opportunistic [16]. Monitoring the prevalence and extent of pathogenic outbreaks is easily performed by PCR using environmental samples [17,18,19]. However, this information may not reveal the potential for infectivity, transmission, or immunological responses over time.