Hematological changes in Florida pompano (Trachinotus carolinus) supplemented with ß-glucan and Pediococcus acidilactici synbiotic.

Florida pompano (Trachinotus carolinus) are a species of growing interest for commercial aquaculture. Effective health monitoring is crucial to the successful growout of the species, and prophylactic and therapeutic use of chemicals and antibiotics has been the traditional strategy for promoting stock health. However, concerns about antimicrobial resistance, chemical residues in seafood products and the environment, and resultant immunosuppression have prompted the industry to identify alternative management strategies, including supplementation with prebiotics, probiotics, and combinations of both (synbiotics). The objectives of this study are to determine and compare hematological, plasma biochemical, and plasma protein electrophoresis data of synbiotic-supplemented (ß-glucan and Pediococcus acidilactici) and non-supplemented Florida pompano. Reference intervals for blood analytes are provided for both groups and for subgroups (females, males, large, and small fish) where statistically significant results exist. There are no differences between the hematological and plasma biochemistry analytes between the supplemented and control groups, except for blood urea nitrogen and carbon dioxide, indicating a possible effect of synbiotic supplementation on gill function and osmoregulation. Sex-related and size-related differences are observed within each of the control and supplemented groups; however, biometric measurements do not strongly correlate with blood analytes. These data represent baseline hematological and plasma biochemical data in the Florida pompano and indicate the safety of synbiotic supplementation in this commercially important species. This study serves to further the commercialization of Florida pompano by providing blood analyte reference intervals for health monitoring in the aquaculture setting.

Diagnosis of piscine francisellosis in Largemouth Bass from a public display exhibit in north-central Florida, USA.

OBJECTIVE: The Largemouth Bass Micropterus salmoides is an important freshwater fish that is native to the southeastern United States and is cultured for conservation, food, and for the sports fishing industry. Francisella orientalis is a globally distributed bacterial pathogen of warmwater fish species and is associated with granulomatous inflammation and high mortalities. Outbreaks of piscine francisellosis in the United States have been reported in only a few fish species. This study describes three case presentations of francisellosis in Largemouth Bass from a public display system in north-central Florida. Additionally, laboratory-controlled immersion challenges using an F. orientalis isolate from tilapia Oreochromis spp. evaluate susceptibility of Largemouth Bass fingerlings to F. orientalis infection and mortality through this exposure route. METHODS: Necropsy, histologic examination, immunohistochemistry, bacterial recovery and culture, and quantitative polymerase chain reaction were used as diagnostic tools to evaluate both the affected display fish and the immersion-challenged fingerlings. RESULT: Although the display fish and immersion-challenged fingerlings presented with nonspecific clinical signs, gross and histological changes were indicative of granulomatous disease. Immunohistochemical and molecular testing methods confirmed F. orientalis infection in affected fish. CONCLUSION: The three case presentations described here mark the first reporting of naturally occurring piscine francisellosis in Largemouth Bass that were held in a public display exhibit. Additionally, causality was proven in the Largemouth Bass fingerlings through the immersion challenges. These findings demonstrate susceptibility through immersion-based exposure and assert that francisellosis should be considered among the list of differential diagnoses for Largemouth Bass with granulomatous disease.

Spatiotemporal patterns in the prevalence of microscopic hepatic changes in Gulf of Mexico Tilefish (Lopholatilus chamaeleonticeps) and associations with hepatic PAHs.

Following the 2010 Deepwater Horizon blowout, demersal longline surveys were conducted across the Gulf of Mexico (GoM) continental shelf to evaluate polycyclic aromatic hydrocarbon (PAH) exposure, tissue accumulation, and health indices in demersal fishes. Tilefish (Lopholatilus chamaeleonticeps), a target species due to Gulf-wide distribution with documented high exposure to PAHs, were collected in the north central GoM at repeat stations 2012 to 2015, and from the northwest GoM, Bay of Campeche, and Yucatán Shelf in 2015 and 2016. Liver samples (n = 239) were analyzed for microscopic hepatic changes (MHCs) by a board-certified veterinary pathologist. Histological analyzes identified 14 MHCs. Prevalence of MHCs was generally uniform throughout the GoM, except for low prevalence on the Yucatán Shelf. Inflammatory and vacuolar changes were most prevalent, while pre-neoplasia and neoplasia were rare. Tilefish sampled annually in the north central GoM showed increases in inflammatory MHCs and glycogen-type vacuolar change over time, while lipid-type vacuolar change decreased over time. Short-term exposure to PAHs was assessed by measuring PAH metabolites in bile (n = 100) using high performance liquid chromatography with fluorescence detection. Longer-term accumulation of PAHs in tissue was assessed by analyzing liver (n = 111) for PAHs and alkylated homologs using QuEChERS extractions and gas chromatography tandem mass spectrometry. Six MHCs including glycogen-type vacuolar change, biliary fibrosis, foci of cellular alteration, parasites, hepatocellular atrophy, and necrosis were significantly associated with hepatic PAH accumulation in Tilefish from the northern central GoM; however, no MHCs were associated with biliary PAH metabolites. Combined with previous studies of PAH exposure and health indices in north central GoM Tilefish post-Deepwater Horizon, which also identified decreases in hepatic lipid storage and Fulton's condition factor that were correlated to increasing PAH exposure, these data indicate concerning temporal trends and changes in hepatic energy storage.

Detection of long chain per- and polyfluoroalkyl substances (PFAS) in the benthic Golden tilefish (Lopholatilus chamaeleonticeps) and their association with microscopic hepatic changes.

Oceans are major sinks for anthropogenic pollutants, including per- and polyfluoroalkyl substances (PFAS). Although PFAS have been detected in surface waters globally, this is the first report of PFAS in a deep (170-400 m) demersal species in the Gulf of Mexico (GoM). Golden Tilefish (Lopholatilus chamaeleonticeps) plasma extracts (n = 185) were investigated for the presence of PFAS using ultra-high performance liquid chromatography-tandem mass spectrometry. A subset of liver tissues (n = 51) were also analyzed for microscopic hepatic changes (MHCs). Overall, nine of the 110 PFAS targeted were detected in Tilefish plasma at relatively high frequencies. Plasma concentrations of total PFAS (Σ9PFAS) ranged from below the detection limit to 27.9 ng g-1 w.w. Significant regional differences were observed with the highest concentrations of PFAS detected in the north central region of the GoM, where substantial industrialization and discharges from the Mississippi River occur. Compared to most wildlife and matrices analyzed globally, the PFAS profiles in Tilefish were unique as they are dominated by PFUnDA. Profile differences are hypothesized to be the result of Tilefish's distinctive lifestyle, habitat, diet, and partitioning characteristics of long-chain PFAS. Several MHCs were identified in this subset of Tilefish that could be detrimental to their health. Significant correlations between PFAS concentrations and biometric indices and MHCs were evident, however, additional research is needed to investigate the role PFAS and PFAS combined with chemical admixtures may play in inducing observed hepatic changes and other physiological effects in Tilefish. These findings give insight into the fate of PFAS at depth in aquatic ecosystems and are cause for concern regarding the health of other deep water benthic biota in GoM and other deepwater sinks for PFAS.

A Retrospective Study of Pathology in Bats Submitted to an Exotic and Zoo Animal Diagnostic Service in Georgia, USA (2008-2019).

Pathology records of bats submitted to the University of Georgia from managed care settings were reviewed to identify naturally occurring diseases. Fifty-nine cases were evaluated during an 11-year period (2008-2019), including representatives from four families: Pteropodidae (Yinpterochiroptera), Phyllostomidae, Vespertilionidae and Molossidae (Yangochiroptera). Pathology reports were reviewed to determine the primary pathological process resulting in death or the decision to euthanize. Cases were categorized as non-infectious (34/59; 58%), infectious/inflammatory (17/59; 29%) or undetermined due to advanced autolysis (8/59; 14%). Musculoskeletal diseases and reproductive losses were the most frequent pathological processes. Among the infectious processes identified, bacterial infections of the reproductive and haemolymphatic systems were most frequently observed. The first two reports of neoplasia in small flying foxes (Pteropus hypomelanus) are described. Bats under managed care present with a wide range of histopathological lesions. In this cohort, non-infectious disease processes were common.

Hepatobiliary PAHs and prevalence of pathological changes in Red Snapper.

Red Snapper (Lutjanus campechanus) were collected throughout the Gulf of Mexico (GoM) from 2011 to 2017 and analyzed for biliary (n = 496) fluorescent aromatic compounds (FACs), hepatic (n = 297) polycyclic aromatic hydrocarbons (PAHs) and microscopic hepatobiliary changes (MHC, n = 152). Gross and histological evaluations were conducted with liver tissues to identify and characterize pathological changes. This is the first report to interrelate hepatobiliary PAH concentrations and MHCs in Red Snapper. Hepatic PAHs measured in GoM Red Snapper ranged from 192 to 8530 ng g-1 w.w. and biliary FACs ranged from 480 to 1,100,000 ng FAC g-1 bile. Biliary FACs in Red Snapper collected along the west Florida Shelf and north central region declined after 2011 and were relatively stable until a sharp increase was noted in 2017. Increases in the PAH exposures are likely due to a number of sources including leaking infrastructure, annual spills, riverine input and the resuspension of contaminated sediments. In contrast, hepatic PAH concentrations were relatively stable indicating Red Snapper are able to maintain metabolic clearance however this energetic cost may be manifesting as microscopic hepatic changes (MHCs). Virtually all (99 %) of the evaluated Red Snapper had one to nine MHCs with an average of five coinciding changes in an individual fish. The observed changes were broadly classified as inflammatory responses, metabolic responses, degenerative lesions, nonneoplastic proliferation and neoplastic lesions. Biliary FACs were associated with parasitic infection and intracellular breakdown product accumulation such as intra-macrophage hemosiderin, lipofuscin and ceroid laden prevalence. Whereas, hepatic PAHs were associated with increased myxozoan plasmodia prevalence. This study evaluates relationships between hepatobiliary PAH concentrations and biometrics, somatic indices, condition factors and microscopic hepatic changes in Red Snapper located in the north central GoM. Together, these results may be signaling increased disease progression in Gulf of Mexico Red Snapper more than likely resulting from chronic environmental stressors including elevated PAH exposures and concentrations.

A histological evaluation of pansteatitis-affected Mozambique tilapia, Oreochromis mossambicus (Peters 1852), from different geographical locations in South Africa.

Pansteatitis is the leading cause for the decline in Nile crocodile populations and the sporadic mortality of fish in the Olifants River System, South Africa. To determine the prevalence of this disease in lentic systems, Mozambique tilapia, Oreochromis mossambicus, were collected from Lake Loskop, Lake Flag Boshielo, Phalaborwa Barrage and Lake Luphephe-Nwanedi. The former three impoundments are located within the main stem of the Olifants River, while the latter, which is geographically isolated and situated in the Limpopo River System, served as a reference site. Mesenteric adipose, liver, serosa of the swim bladder, gill and the skeletal muscle of fish sampled were examined for gross and microscopic evidence of pansteatitis. Microscopically observed changes were used to statistically compare pansteatitis prevalence between samples and sites. Based on histopathological evaluation, the adipose tissue in the liver, swim bladder serosa and coelom from severely debilitated individuals showed the most significant pathological changes. Lesions indicative of steatitis were observed in fish collected from Lake Loskop (75%), Lake Flag Boshielo (22%) and Lake Luphephe-Nwanedi (15%). Further investigation is warranted to understand the pervasiveness and mechanisms driving pathological changes of pansteatitis at Lake Flag Boshielo, Phalaborwa Barrage and Lake Luphephe-Nwanedi.

Longitudinal and Cross-Sectional Sampling of Serpentovirus (Nidovirus) Infection in Captive Snakes Reveals High Prevalence, Persistent Infection, and Increased Mortality in Pythons and Divergent Serpentovirus Infection in Boas and Colubrids.

The aim of this study of serpentovirus infection in captive snakes was to assess the susceptibility of different types of snakes to infection and disease, to survey viral genetic diversity, and to evaluate management practices that may limit infection and disease. Antemortem oral swabs were collected from 639 snakes from 12 US collections, including 62 species, 28 genera, and 6 families: Pythonidae (N = 414 snakes; pythons were overrepresented in the sample population), Boidae (79), Colubridae (116), Lamprophiidae (4), Elapidae (12), and Viperidae (14). Infection was more common in pythons (38%; 95% CI: 33.1-42.4%), and in boas (10%; 95% CI: 5.2-18.7%) than in colubrids (0.9%, 95% CI: <0.01-4.7%); infection was not detected in other snake families (lamprophiids 0/4, 95% CI: 0-49%; elapids 0/12, 95% CI: 0-24.2%; and vipers 0/14, 95% CI: 0-21.5%), but more of these snakes need to be tested to confirm these findings. Clinical signs of respiratory disease were common in infected pythons (85 of 144). Respiratory signs were only observed in 1 of 8 infected boas and were absent in the single infected colubrid. Divergent serpentoviruses were detected in pythons, boas, and colubrids, suggesting that different serpentoviruses might vary in their ability to infect snakes of different families. Older snakes were more likely to be infected than younger snakes (p-value < 0.001) but males and females were equally likely to be infected (female prevalence: 23.4%, 95% CI 18.7-28.9%; male prevalence: 23.5%, 95% CI 18-30.1%; p-value = 0.144). Neither age (p-value = 0.32) nor sex (p-value = 0.06) was statistically associated with disease severity. Longitudinal sampling of pythons in a single collection over 28 months revealed serpentovirus infection is persistent, and viral clearance was not observed. In this collection, infection was associated with significantly increased rates of mortality (p-value = 0.001) with death of 75% of infected pythons and no uninfected pythons over this period. Offspring of infected parents were followed: vertical transmission either does not occur or occurs with a much lower efficiency than horizontal transmission. Overall, these findings confirm that serpentoviruses pose a significant threat to the health of captive python populations and can cause infection in boa and colubrid species.

Diagnosing Microcystin Intoxication of Canines: Clinicopathological Indications, Pathological Characteristics, and Analytical Detection in Postmortem and Antemortem Samples.

In the summer of 2018, six dogs exposed to a harmful algal bloom (HAB) of Microcystis in Martin County Florida (USA) developed clinicopathological signs of microcystin (MC) intoxication (i.e., acute vomiting, diarrhea, severe thrombocytopenia, elevated alanine aminotransferase, hemorrhage). Successful supportive veterinary care was provided and led to survival of all but one patient. Confirmation of MC intoxication was made through interpretation of clinicopathological abnormalities, pathological examination of tissues, microscopy (vomitus), and analytical MC testing of antemortem/postmortem samples (vomitus, blood, urine, bile, liver, kidney, hair). Gross and microscopic examination of the deceased patient confirmed massive hepatic necrosis, mild multifocal renal tubular necrosis, and hemorrhage within multiple organ systems. Microscopy of a vomitus sample confirmed the presence of Microcystis. Three analytical MC testing approaches were used, including the MMPB (2-methyl-3-methoxy-4-phenylbutyric acid) technique, targeted congener analysis (e.g., liquid chromatography tandem-mass spectrometry of MC-LR), and enzyme-linked immunosorbent assay (ELISA). Total Adda MCs (as MMPB) were confirmed in the liver, bile, kidney, urine, and blood of the deceased dog. Urinalysis (MMPB) of one surviving dog showed a high level of MCs (32,000 ng mL-1) 1-day post exposure, with MCs detectable >2 months post exposure. Furthermore, hair from a surviving dog was positive for MMPB, illustrating another testable route of MC elimination in canines. The described cases represent the first use of urine as an antemortem, non-invasive specimen to diagnose microcystin toxicosis. Antemortem diagnostic testing to confirm MC intoxication cases, whether acute or chronic, is crucial for providing optimal supportive care and mitigating MC exposure.

Lipidomics for wildlife disease etiology and biomarker discovery: a case study of pansteatitis outbreak in South Africa.

INTRODUCTION: Lipidomics is an emerging field with great promise for biomarker and mechanistic studies due to lipids diverse biological roles. Clinical research applying lipidomics is drastically increasing, with research methods and tools developed for clinical applications equally promising for wildlife studies. OBJECTIVES: Limited research to date has applied lipidomics, especially of the intact lipidome, to wildlife studies. Therefore, we examine the application of lipidomics for in situ studies on Mozambique tilapia (Oreochromis mossambicus) in Loskop Dam, South Africa. Wide-scale mortality events of aquatic life associated with an environmentally-derived inflammatory disease, pansteatitis, have occurred in this area. METHODS: The lipidome of adipose tissue (n = 31) and plasma (n = 51) from tilapia collected from Loskop Dam were characterized using state of the art liquid chromatography coupled to high-resolution tandem mass spectrometry. RESULTS: Lipid profiles reflected pansteatitis severity and were significantly different between diseased and healthy individuals. Over 13 classes of lipids associated with inflammation, cell death, and/or oxidative damage were upregulated in pansteatitis-affected adipose tissue, including ether-lipids, short-chained triglyceride oxidation products, sphingolipids, and acylcarnitines. Ceramides showed a 1000-fold increase in the most affected adipose tissues and were sensitive to disease severity. In plasma, triglycerides were found to be downregulated in pansteatitis-affected tilapia. CONCLUSION: Intact lipidomics provided useful mechanistic data and possible biomarkers of pansteatitis. Lipids pointed to upregulated inflammatory pathways, and ceramides serve as promising biomarker candidates for pansteatitis. As comprehensive coverage of the lipidome aids in the elucidation of possible disease mechanisms, application of lipidomics could be applied to the understanding of other environmentally-derived inflammatory conditions, such as those caused by obesogens.

Variation among human, veterinary and environmental Mycobacterium chelonae-abscessus complex isolates observed using core genome phylogenomic analysis, targeted gene comparison, and anti-microbial susceptibility patterns.

Mycobacterium chelonae is a member of the Mycobacterium chelonae-abscessus complex and a cause of opportunistic disease in fish, reptiles, birds, and mammals including humans. Isolates in the complex are often difficult to identify and have differing antimicrobial susceptibilities. Thirty-one previously identified rapidly-growing, non-tuberculous Mycobacterium sp. isolates cultured from biofilms, fish, reptiles, mammals, including humans, and three ATCC reference strains were evaluated with nine M. chelonae-abscessus complex whole genome sequences from GenBank by phylogenomic analysis, targeted gene comparisons, and in-vitro antimicrobial susceptibility patterns to assess strain variation among isolates from different sources. Results revealed minimal genetic variation among the M. chelonae strains. However, the core genomic alignment and SNP pattern of the complete 16S rRNA sequence clearly separated the turtle type strain ATCC 35752T from the clinical isolates and human reference strain "M. chelonae chemovar niacinogenes" ATCC 19237, providing evidence of two distinct subspecies. Concatenation of the partial rpoB (752 bp) and complete hsp65 (1,626 bp) sequence produced the same species/subspecies delineations as the core phylogeny. Partial rpoB and hsp65 sequences identified all the clinical isolates to the appropriate species level when respective cut-offs of 98% and 98.4% identity to the M. chelonae type strain ATCC 35752T were employed. The human strain, ATCC19237, was the most representative strain for the evaluated human, veterinary, and environmental strains. Additionally, two isolates were identified as Mycobacterium saopaulense, its first identification in a non-fish or non-human host.

Mycobacterium syngnathidarum sp. nov., a rapidly growing mycobacterium identified in syngnathid fish.

Two closely related isolates, 27335T and 24999, of rapidly growing, non-pigmented mycobacteria, were cultured from two clinically ill fish of the family Syngnathidae. Whole genome sequencing of the two isolates revealed low sequence homology to documented mycobacteria within public databases such as the NCBI. Evaluation of targeted housekeeping genes, including 16S rRNA, ITS, rpoB and hsp65, related the two bacteria distantly to Mycobacterium senegalense CK2 M4421 and Mycobacterium farcinogenes DSM 43637. Phenotypic, biochemical and dDNA-DNA hybridization tests demonstrated that Mycobacterium syngnathidarum is a new species distinct from other recognized rapidly growing mycobacterial species. Phenotypic, chemotaxonomic and phylogenetic data evaluation provided evidence that the two strains represent one novel species. We propose the formal recognition of Mycobacterium syngnathidarum sp. nov., with isolate 27335T as the type strain (=ATCC TSD-89T,=DSM 105112T).

BAYLISASCARIS PROCYONIS LARVA MIGRANS IN TWO CAPTIVE NORTH AMERICAN BEAVERS (CASTOR CANADENSIS).

Baylisascaris procyonis larva migrans was diagnosed in two North American beavers ( Castor canadensis ) belonging to a zoological park in Clarke County, Georgia. Both beavers presented with neurological signs. One beaver died naturally and despite attempted treatment, the other beaver was euthanatized because of severe clinical signs and poor prognosis. Histologic evaluation of the beavers revealed evidence of parasitic migration characterized by several lesions, including eosinophilic granulomas in various organs, as well as necrotizing eosinophilic and lymphoplasmacytic to granulomatous polioencephalitis, leukoencephalitis and cervical leukomyelitis. This represents the first confirmed case of B. procyonis larva migrans in beaver and the first non-raccoon ( Procyon lotor ) host in the southeastern United States. This report highlights the need for clinicians and diagnosticians to consider baylisascariasis in animals with compatible clinical signs. Preventative measures should be considered for captive animals, because early diagnosis of B. procyonis is challenging, and treatment is often unrewarding.

Histologic and molecular characterization of Edwardsiella piscicida infection in largemouth bass (Micropterus salmoides).

The genus Edwardsiella is composed of a diverse group of facultative anaerobic, gram-negative bacteria that can produce disease in a wide variety of hosts, including birds, reptiles, mammals, and fish. Our report describes the isolation and identification of Edwardsiella piscicida associated with chronic mortality events in 2 separate captive largemouth bass (Micropterus salmoides) populations in New York and Florida. Wet-mount biopsies of skin mucus, gill, kidney, and spleen from several affected largemouth bass contained significant numbers of motile bacteria. Histologic examination revealed multifocal areas of necrosis scattered throughout the heart, liver, anterior kidney, posterior kidney, and spleen. Many of the necrotic foci were encapsulated or replaced by discrete granulomas and associated with colonies of gram-negative bacteria. Initial phenotypic and matrix-assisted laser desorption ionization-time of flight mass spectrometric analysis against existing spectral databases of recovered isolates identified these bacteria as Edwardsiella tarda Subsequent molecular analysis using repetitive sequence mediated and species-specific PCR, as well as 16S rRNA, rpoB, and gyrB sequences, classified these isolates as E. piscicida As a newly designated taxon, E. piscicida should be considered as a differential for multiorgan necrosis and granulomas in largemouth bass.

Disseminated Yersinia pseudotuberculosis infection in a paca (Cuniculus paca).

A 2-yr-old paca (Cuniculus paca) was presented for necropsy with a history of sudden death. GrosS examination revealed multifocal, transmural, well-demarcated, white, soft nodules scattered along the length of the small intestine. The liver also had similar nodules associated with the capsular and cut surface. Histologic evaluation of several organs, including the intestine, liver, lung, kidney, adrenal gland, and lymph nodes, was consistent with disseminated yersiniosis. In addition, aerobic bacterial culture of liver and lung tissue yielded heavy growth of Yersinia pseudotuberculosis. Yersinia pseudotuberculosis is a Gram-negative, enteric pathogen that can cause disease in a variety of terrestrial species including humans. Although systemic infection has been observed in rodent species, to our knowledge this is the first report of disseminated Y pseudotuberculosis in a paca.