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OBJECTIVE: The purpose of this study was to compare various sampling techniques and commercially available diagnostic tests for Cryptosporidium serpentis. METHODS: A colony of 80 eastern indigo snakes (Drymarchon couperi) in human care was screened for the presence of C serpentis using endoscopic gastric mucosal biopsies for histologic and molecular analyses. At the time of endoscopic examination and biopsy, a cloacal swab, gastric swab, and gastric lavage sample were also collected. A C serpentis-specific probe hybridization quantitative PCR (qPCR) was performed on each sample. The gastric lavage sample was divided equally for direct microscopy, acid-fast stain, rapid qualitative immunochromatographic assay, direct fluorescent antibody, and 5 different PCR analyses. If a fecal sample was available at the time of endoscopic evaluation, it was also evaluated for Cryptosporidium oocysts by direct microscopy and acid-fast staining. RESULTS: When comparing test results to histologic analyses, the sensitivity of the probe hybridization qPCR of gastric biopsy, gastric lavage, and gastric swab was 100% while the cloacal swab was 72%. When gastric lavage tests were compared, qPCRs outperformed the other tests. CONCLUSIONS: Endoscopic biopsy for histologic and qPCR analyses is recommended for disease diagnosis, while gastric lavage or gastric swab samples for qPCR analysis are as sensitive as endoscopic biopsy for screening for the pathogen but cannot diagnose disease. CLINICAL RELEVANCE: The results from this study allow the veterinary practitioner to select the most appropriate sample and testing methodology when evaluating an ophidian patient for gastric cryptosporidiosis.
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There is a paucity of information regarding the health status of free-ranging eastern indigo snakes (EIS; Drymarchon couperi) in heavily modified and developing landscapes. As a component of regional Florida Everglades restoration efforts, several areas occupied by EIS are being converted from agricultural lands to reservoirs. From 2020 to 2022, 28 EIS were opportunistically captured at two of these sites and brought into captivity to join a captive breeding colony; however, 11 snakes died within 5 mo of capture. Health assessments were performed on 28 individuals and included hematology and plasma biochemistry analysis, as well as screening for pesticide contaminant levels, parasites, and other pathogens. Overall, the presence of pathogens was relatively high, suggesting immunosuppression secondary to stress: 25/28 (89.4%) Kalicephalus sp.; 12/28 (42.9%) Raillietiella orientalis; 11/28 (39.2%) Ochetosoma validum; 7/28 (25.0%) Cryptosporidium serpentis; 3/28 (10.7%) snake adenovirus 1; and 1/28 (3.6%) Ferlavirus genotype C. Stress may have been caused by physical displacement, habitat modification, and noise pollution. These potential stressors (including the presence of remnant harmful chemicals from previous land use and the impacts on this federally threatened species) should be considered further when making restoration or construction decisions.
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Colubridae , Criptosporidiose , Cryptosporidium , Humanos , Animais , Florida/epidemiologia , Serpentes , EcossistemaRESUMO
Madagascar hissing cockroaches (MHC, Gromphadorhina portentosa) are members of the Blaberidae (giant cockroaches) family of the Insecta class. They are native to the African island of Madagascar where they live within leaf litter on the rainforest floor. Due to their large size, relative tameness, and general easy keeping, they have become popular in classrooms, zoological collections, museums, research laboratories, and as private exotic pets; however, descriptions of diseases of MHC in the literature are rare. The objective of this study is to describe and characterize postmortem histological findings in 18 captive MHC from a single zoological collection. In this retrospective study, 18 (4 females and 14 males) adult MHC necropsies were submitted to Northwest ZooPath between 2016 and 2020 for evaluation. The main organs with histological lesions were chitinous gut (foregut and/or hindgut; n = 17), tracheae (n = 15), fat body (n = 14), ventriculus (midgut) (n = 13), body wall (n = 12), Malpighian tubules (n = 12), and hemolymphatic sinuses (n = 12). All animals had inflammatory lesions affecting various organs. Inflammatory lesions typically consisted of aggregates of hemocytes with variable amounts of melanization and/or encapsulation. Bacterial, fungal, and parasitic infections were common and variably associated with hemocytic inflammation. Many of these organisms may represent symbiotic organisms of the MHC that cause opportunistic infections. This study contributes to the current knowledge of pathological findings and disease response of MHC and reviews diseases reported in multiple cockroach species.
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Baratas , Masculino , Feminino , Animais , Baratas/microbiologia , Baratas/fisiologia , Madagáscar , Estudos RetrospectivosRESUMO
Erythrocyte sedimentation rate (ESR) is a hematological test that can detect inflammatory activity within the body. Although not specific for any particular disease, ESR is often used as a screening "sickness indicator" due to its reliability and low cost. The Westergren method is a manual ESR technique commonly used but requires special graduated pipettes and over 1mL of whole blood, precluding its use in smaller patients where limited sample volumes can be obtained. A modified micro-ESR technique has been described using hematocrit capillary tubes but is used less commonly. ESR has been reported to be a useful inflammatory indicator in gopher tortoises (Gopherus polyphemus) and box turtles (Terrapene spp.) but not in Florida cottonmouth snakes (Agkistrodon conanti). Having an inexpensive screening test for inflammation can help guide medical decisions within conservation efforts of imperiled species. This study evaluated the correlation between these two ESR methodologies in threatened eastern indigo snakes (Drymarchon couperi, EIS) and found a very strong correlation (rs = 0.897), without constant or proportional biases and a reference interval of 0 (90% CI -1-1)-9 mm/h (90% CI 8-11) was defined. Additionally, a significant difference was found between healthy EIS and EIS in mid-ecdysis (p = 0.006) and EIS with gastric cryptosporidiosis (p = 0.006), indicating ESR as a useful inflammatory indicator in EIS.
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Understanding risk factors associated with reintroductions is important for making informed decisions within an adaptive framework. Biosecurity measures minimizing the risk of the introduction or spread of transmissible diseases are a priority when considering the release of captive-reared wildlife. Eastern indigo snake (EIS; Drymarchon couperi) reintroductions have been occurring in Alabama since 2010 and in Florida since 2017. During this effort the pathogen Cryptosporidium serpentis was detected, affecting several of the captive breeding snakes. Infected snakes were quarantined and removed from breeding efforts, which reduced snakes available for the reintroduction projects. To make informed management decisions about future reintroduction strategies, 155 free-ranging snakes were sampled at the two release sites and a third site in Georgia to evaluate the natural occurrence of C. serpentis. Additionally, 72 free-ranging EIS and other species incidentally encountered throughout the EIS range were tested opportunistically. All snakes sampled at the three focal sites tested negative, but one opportunistically tested EIS from South Florida tested positive. These results indicate that C. serpentis is present in the environment in at least one location, but at low levels. Our results suggest that, pending additional surveillance, C. serpentispositive snakes should not be included in reintroduction efforts, and that maintaining a high level of biosecurity is important in captive breeding programs.
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Colubridae , Criptosporidiose , Cryptosporidium , Animais , Criptosporidiose/epidemiologia , Serpentes , Animais Selvagens , GeorgiaRESUMO
A probe-hybridization quantitative polymerase chain reaction assay specific for Cryptosporidium serpentis (qPCR) has been developed and shown to be extremely sensitive in the laboratory, but clinical sensitivity and specificity for this test are lacking. To approximate the sensitivity and specificity of the C. serpentis qPCR, the medical records from a captive snake colony were reviewed, and between November 2015 and June 2021, 63 eastern indigo snakes (Drymarchon couperi) were necropsied. Of these 63 snakes, 11 had qPCR performed on gastric biopsies collected at the time of necropsy, 8 had qPCR performed on samples collected by gastric swab within 35 days of necropsy, and 34 had qPCR performed on samples collected by cloacal swab within 84 days of necropsy. The qPCR results were then compared to the post-mortem histological findings, where all three sampling techniques had a 100% specificity. The sensitivity was highest in samples collected at necropsy (100%, CI: 63.06 - 100%) followed by the ante-mortem testing: gastric swab (87.50%, CI: 42.13 - 99.64%) and cloacal swab (66.67%, CI: 44.68 - 84.37%).
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Colubridae , Criptosporidiose , Cryptosporidium , Animais , Humanos , Cryptosporidium/genética , Serpentes , Reação em Cadeia da Polimerase em Tempo Real , EstômagoRESUMO
Medical and husbandry records of eastern indigo snakes (EIS) (Drymarchon couperi) housed at the Orianne Center for Indigo Conservation (OCIC) were reviewed to determine risk factors for developing dystocia. Thirty-four cases of dystocia were identified out of 104 successful breeding events between 2010 and 2020. The number of breeding events, age, body weight, housing, and selected blood parameters were reviewed for the female EIS successfully bred at OCIC between 2010 and 2020. Categorical data were evaluated with chi-square and the Fisher exact test, and the continuous data were evaluated using analysis of variance. Differences in data were considered statistically significant when P < 0.05. There was a significant difference seen between EIS that developed dystocia (EISd) and EIS that did not (EISn) for breeding events, with the virgin breeding event having more EISd than any other breeding event. Risk analysis of the data revealed that EIS were 9.28 times more likely to develop dystocia during the virgin breeding event than on subsequent breeding events. Age and housing were not significantly different between EISd and EISn for each breeding event, but body weight was, as EIS weighing less than 1.5 kg were found to be 4.43 times more likely to develop dystocia. There was also a significant difference between EISd and EISn in plasma sodium, calcium, ionized calcium, glucose, and albumin. When compared to EISn, EISd had lower plasma sodium, calcium, ionized calcium, and albumin and higher plasma glucose. There were also significant differences in the white blood cell differential count, as EISd had lower basophils and monocytes when compared to EISn. The majority of EISd were managed surgically, and retained eggs were more likely to be located within the right oviduct. This retrospective study is the first to evaluate potential causes of dystocia in EIS.
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Colubridae , Distocia/veterinária , Envelhecimento , Criação de Animais Domésticos , Animais , Conservação dos Recursos Naturais , Espécies em Perigo de Extinção , Feminino , Gravidez , Estudos RetrospectivosRESUMO
When organisms possess chemical defenses, their predators may eventually evolve resistance to their toxins. Eastern indigo snakes (Drymarchon couperi; EIS) prey on pitvipers and are suspected to possess physiological resistance to their venom. In this study, we formally investigated this hypothesis using microassays that measured the ability of EIS blood sera to inhibit (A) hemolytic and (B) snake venom metalloproteinase (SVMP) activity of copperhead (Agkistrodon contortrix) venom. To serve as controls, we also tested the inhibitory ability of sera from house mice (Mus musculus) and checkered gartersnakes (Thamnophis marcianus), a snake that does not feed on pitvipers. Sera from both EIS and gartersnakes inhibited over 60% of SVMP activity, while only EIS sera also inhibited venom hemolytic activity (78%). Our results demonstrate that EIS serum is indeed capable of inhibiting two of the primary classes of toxins found in copperhead venom, providing the first empirical evidence suggesting that EIS possess physiological resistance to venom upon injection. Because we documented resistance to hemolytic components of pitviper venom within EIS but not gartersnakes, we speculate this resistance may be driven by selection from feeding on pitvipers while resistance to SVMP may be relatively widespread among snakes.
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While practicing exotic animal medicine as an ambulatory practitioner, veterinarians need to be prepared for the inevitable emergency call. Emergencies in exotic animal medicine come in all shapes and sizes and the veterinarian must be prepared for a variety of situations. With the proper training, equipment, and managing client expectations, an ambulatory exotics animal practitioner can successfully address emergencies. This article provides a brief overview in managing emergency cases in an ambulatory exotics animal practice.