FOUR CASES OF SEVERED PEDUNCLES IN BOTTLENOSE DOLPHINS (TURSIOPS TRUNCATUS) ALONG THE ALABAMA COAST.

Alabama (AL) is a hotspot in the Gulf of Mexico (GoM) for human interaction-related cetacean strandings, including harassment, vessel strikes, and fisheries interactions. We examined four bottlenose dolphins (Tursiops truncatus) stranded dead along the AL coast during 2012-2017 with severed peduncles suspected to be related to human interaction (HI). Evidence from each case, including photographs, gross necropsy results, and histopathologic findings when available, was reviewed to determine the mode of severance and whether it contributed to death. In each case, the severance site had smooth, clean edges on at least one side, indicating the use of a sharp instrument to remove the caudal peduncle and flukes. Three cases also had evidence of fisheries interactions, including linear impressions around the rostrum, fins and/or flukes, indicating that these animals may have been entangled in fisheries gear prior to death. Histopathology in one of these cases revealed that the severance occurred perimortem; speculatively, the caudal peduncle and flukes may have been cut off to facilitate removing the dolphin from its entanglement. Although cases of amputation and mutilation are not uncommon globally among stranding reports, few cases have been described and analyzed in the literature. This paper is the first to document and compare multiple cases of severed peduncles with evidence of HI, including fisheries, in the GoM. This case series enhances our understanding of the types of HI occurring in bottlenose dolphins and highlights the need for continued public education, policy, and management to address cases like these.

The Gombe Ecosystem Health Project: 16 years of program evolution and lessons learned.

Infectious disease outbreaks pose a significant threat to the conservation of chimpanzees (Pan troglodytes) and all threatened nonhuman primates. Characterizing and mitigating these threats to support the sustainability and welfare of wild populations is of the highest priority. In an attempt to understand and mitigate the risk of disease for the chimpanzees of Gombe National Park, Tanzania, we initiated a long-term health-monitoring program in 2004. While the initial focus was to expand the ongoing behavioral research on chimpanzees to include standardized data on clinical signs of health, it soon became evident that the scope of the project would ideally include diagnostic surveillance of pathogens for all primates (including people) and domestic animals, both within and surrounding the National Park. Integration of these data, along with in-depth post-mortem examinations, have allowed us to establish baseline health indicators to inform outbreak response. Here, we describe the development and expansion of the Gombe Ecosystem Health project, review major findings from the research and summarize the challenges and lessons learned over the past 16 years. We also highlight future directions and present the opportunities and challenges that remain when implementing studies of ecosystem health in a complex, multispecies environment.

Brucella ceti sequence type 23, 26, and 27 infections in North American cetaceans.

Brucella ceti infection is associated with a variety of disease outcomes in cetaceans globally. Multiple genotypes of B. ceti have been identified. This retrospective aimed to determine if specific lesions were associated with different B. ceti DNA sequence types (STs). Characterization of ST was performed on 163 samples from 88 free-ranging cetaceans, including common bottlenose dolphin Tursiops truncatus (T.t.; n = 73), common short-beaked dolphin Delphinus delphis (D.d.; n = 7), striped dolphin Stenella coeruleoalba (n = 3), Pacific white-sided dolphin Lagenorhynchus obliquidens (n = 2), sperm whale Physeter macrocephalus (n = 2), and harbour porpoise Phocoena phocoena (n = 1), that stranded along the coast of the US mainland and Hawaii. ST was determined using a previously described insertion sequence 711 quantitative PCR. Concordance with 9-locus multi-locus sequence typing was assessed in a subset of samples (n = 18). ST 26 was most commonly identified in adult dolphins along the US east coast with non-suppurative meningoencephalitis (p = 0.009). Animals infected with ST 27 were predominately perinates that were aborted or died shortly after birth with evidence of in utero pneumonia (p = 0.035). Reproductive tract inflammation and meningoencephalitis were also observed in adult T.t. and D.d. with ST 27, though low sample size limited interpretation. ST 23 infections can cause disease in cetacean families other than porpoises (Phocoenidae), including neurobrucellosis in D.d. In total, 11 animals were potentially infected with multiple STs. These data indicate differences in pathogenesis among B. ceti STs in free-ranging cetaceans, and infection with multiple STs is possible.

GLOBAL RETROSPECTIVE ANALYSIS OF PATHOLOGICAL FINDINGS IN ZOO-MANAGED GOELDI'S MONKEYS (CALLIMICO GOELDII), 1965-2018.

As part of the collaborative efforts and goals of managing zoo-housed Goeldi's monkeys, or callimicos (Callimico goeldii), a retrospective review of gross and histopathological postmortem examination reports submitted to the International Studbook Coordinator was carried out by veterinary representatives of the Species Survival Plan to investigate disease trends. A total of 1,887 postmortem reports (1965-2018) collected from more than 150 institutions were reviewed. Histologic findings from 862 postmortem reports and primary causes of mortality were compiled to determine the most common findings. Within the study population, 419 individuals (48.6%) were male, 383 (44.4%) female, and the remaining 60 (7%) of undetermined sex. The primary lesion at death in adults was chronic renal disease. The other prevalent lesions included cardiac disease, myelolipomas, enteritis, colitis, and hepatitis. In Great Britain and mainland Europe, Yersinia spp. infection had significantly higher prevalence than in North American callimico populations. Multiple lesions affecting more than one organ system were identified in many animals of this study population. Results also showed that for adult callimicos in zoological institutions in North America, Europe, and Great Britain, life span has been increasing over the last 50 yr.

Shell Lesions Associated With Emydomyces testavorans Infection in Freshwater Aquatic Turtles.

A newly described onygenalean fungus, Emydomyces testavorans, has been isolated from ulcerative shell and skin lesions of freshwater aquatic chelonians. To investigate the shell lesions associated with infection and determine if any lesional features were unique to E. testavorans, tissues from turtles housed in zoological institutions (n = 45) in the United States and free-living turtles (n = 5) submitted for diagnostic biopsy or necropsy were examined. Free-living turtles were from geographically distinct habitats in Florida (n = 1) and Washington (n = 4) at the time of sampling. Histologic shell sections were evaluated for the presence or absence of specific lesional features. Infection with E. testavorans was evaluated in all cases by screening GMS (Grocott-Gomori's methenamine silver)-stained histologic sections for the presence of morphologically consistent fungi and by quantitative PCR (polymerase chain reaction) on representative frozen tissue or formalin-fixed paraffin-embedded sections. Additionally, culture was performed for 15 cases with available fresh/frozen tissue. In total, there were 17 PCR-confirmed E. testavorans cases, 29 cases with morphologically consistent fungi on GMS-stained sections, and 21 cases of shell lesions without histologic or molecular evidence of E. testavorans infection. Epithelial inclusion cysts, defined as cystic structures within the dermis lined by keratinized stratified squamous epithelium and containing necrotic bone and keratin debris, were significantly (P < .01) associated with E. testavorans infection. Other significantly associated shell lesions included squamous metaplasia, hyperkeratosis, inflammation, and osteonecrosis (P < .05). This study identified characteristic shell lesions associated with E. testavorans infection. Further studies to prove causality are needed.

Novel Herpesvirus Associated With Oropharyngeal Squamous Cell Carcinoma in Smooth Green Snakes (Opheodrys vernalis).

During a 19-month period, 5 smooth green snakes (Opheodrys vernalis) maintained as an ex situ conservation colony presented with rapid clinical progression of locally invasive oropharyngeal squamous cell carcinoma. All 5 originated from the same wild source and were housed together or in close proximity. An infectious cause was considered likely, and nested conventional polymerase chain reaction (PCR) and in situ hybridization confirmed the presence of a novel alphaherpesvirus, Opheodrys herpesvirus 1, in the neoplastic tissue in 4 of the 5 snakes. Retrospective screening of previously submitted smooth green snakes by in situ hybridization did not detect virus in prior submissions from the colony. This report documents molecular characterization of an ophidian herpesvirus as well as colocalization of its viral nucleic acid with neoplastic transformation in snakes.

Oesophagostomiasis in non-human primates of Gombe National Park, Tanzania.

Oesophagostomum sp. is a parasitic nematode that frequently infects wild chimpanzees. Although nodular lesions are commonly associated with infection, some wild chimpanzee populations seem to tolerate Oesophagostomum nodular lesions while those at Gombe and other sites suffer from associated morbidity and mortality. From August 2004 to December 2013, we examined demographic (i.e., age, sex) and individual correlates (i.e., fecal consistency, Oesophagostomum egg production) to Oesophagostomum-associated pathology in 14 individually recognized chimpanzees at Gombe Stream National Park, Tanzania. In addition, we characterized Oesophagostomum-associated pathology in 14 individual sympatric primates including baboons, colobus, and cercopithecid monkeys. In five chimpanzees, there was no evidence of any significant underlying disease aside from oesophagostomiasis to explain the thin condition or diarrhea. All five of these chimpanzees had moderate to numerous parasitic nodules. In general, nodules were more numerous in older chimpanzees. Three of four chimpanzees with the highest average Oesophagostomum egg counts in feces collected during the year prior to their death had numerous parasitic nodules at necropsy. In contrast, the four chimpanzees with the lowest egg counts had only moderate numbers of nodules. No association (P = 0.74) was noted between frequency of diarrhea in the year prior to death and the number of nodules noted at necropsy. Nodules were also present in all baboons examined documenting pathology associated with Oesophagostomum infection in wild baboons. In contrast, no lesions were noted in colobus or cercopithecid monkeys, although it is uncertain if they are infected as no fecal studies have been completed in these species to date at Gombe. Sequence of DNA isolated from nodules in chimpanzees matched (99%) Oesophagostomum stephanostomum. Further research is needed to identify the types of Oesophagostomum causing lesions in baboons and to determine if baboons suffer from these infections. Am. J. Primatol. 80:e22572, 2018. © 2016 Wiley Periodicals, Inc.

CANINE DISTEMPER IN A VACCINATED SNOW LEOPARD ( PANTHERA UNCIA).

A 6-yr-old male snow leopard ( Panthera uncia) presented with acute seizures, hyperthermia, and tachypnea. Because of a diagnosis of anuric renal failure, the animal was euthanized. On histopathologic examination, numerous intralesional intracytoplasmic and intranuclear inclusions were found in the lungs, lymph nodes, and stomach. Positive immunohistochemical staining for canine distemper virus (CDV) was found in the lungs and, to a lesser extent, in the lymph nodes and brain. Molecular testing yielded a CDV H gene sequence that was closely related to CDV isolates concurrently found in wild raccoons from adjacent forested areas. The leopard had been vaccinated once against CDV with the use of a recombinant canarypox-vectored live vaccine during a routine wellness examination 12 wk prior to death. Serial serum neutralization titers performed on banked serum collected between vaccination and death showed poor serologic response to the vaccine. This case demonstrates a probable failure of protection against naturally occurring CDV.

Fetal distress and in utero pneumonia in perinatal dolphins during the Northern Gulf of Mexico unusual mortality event.

An unusual mortality event (UME) involving primarily common bottlenose dolphins Tursiops truncatus of all size classes stranding along coastal Louisiana, Mississippi, and Alabama, USA, started in early 2010 and continued into 2014. During this northern Gulf of Mexico UME, a distinct cluster of perinatal dolphins (total body length <115 cm) stranded in Mississippi and Alabama during 2011. The proportion of annual dolphin strandings that were perinates between 2009 and 2013 were compared to baseline strandings (2000-2005). A case-reference study was conducted to compare demographics, histologic lesions, and Brucella sp. infection prevalence in 69 UME perinatal dolphins to findings from 26 reference perinates stranded in South Carolina and Florida outside of the UME area. Compared to reference perinates, UME perinates were more likely to have died in utero or very soon after birth (presence of atelectasis in 88 vs. 15%, p < 0.0001), have fetal distress (87 vs. 27%, p < 0.0001), and have pneumonia not associated with lungworm infection (65 vs. 19%, p = 0.0001). The percentage of perinates with Brucella sp. infections identified via lung PCR was higher among UME perinates stranding in Mississippi and Alabama compared to reference perinates (61 vs. 24%, p = 0.01), and multiple different Brucella omp genetic sequences were identified in UME perinates. These results support that from 2011 to 2013, during the northern Gulf of Mexico UME, bottlenose dolphins were particularly susceptible to late-term pregnancy failures and development of in utero infections including brucellosis.

CUTANEOUS SQUAMOUS CELL CARCINOMA IN A PANTHER CHAMELEON (FURCIFER PARDALIS) AND TREATMENT WITH CARBOPLATIN IMPLANTABLE BEADS.

A 3-yr-old male panther chameleon (Furcifer pardalis) presented with bilateral raised crusted skin lesions along the lateral body wall that were found to be carcinoma in situ and squamous cell carcinoma. Similar lesions later developed on the caudal body wall and tail. A subcutaneous implantable carboplatin bead was placed in the first squamous cell carcinoma lesion identified. Additional new lesions sampled were also found to be squamous cell carcinomas, and viral polymerase chain reaction was negative for papillomaviruses and herpesviruses. Significant skin loss would have resulted from excision of all the lesions, so treatment with only carboplatin beads was used. No adverse effects were observed. Lesions not excised that were treated with beads decreased in size. This is the first description of cutaneous squamous cell carcinoma and treatment with carboplatin implantable beads in a panther chameleon.

COINFECTION OF CALIFORNIA SEA LION ADENOVIRUS 1 AND A NOVEL POLYOMAVIRUS IN A HAWAIIAN MONK SEAL (NEOMONACHUS SCHAUINSLANDI).

The Hawaiian monk seal (Neomonachus schauinslandi) is an endangered species. Here, we present a clinical case of a 26-yr-old male Hawaiian monk seal (HMS) kept in an aquarium with a history of intermittent anorexia and evidence of renal disease. Histologic examination revealed eosinophilic intranuclear inclusions in the liver. Conventional nested PCR protocols were used to test for viruses, and it tested positive for adenovirus and polyomavirus, and negative for herpesvirus. The adenovirus partial polymerase gene is 100% homologous to that of California sea lion adenovirus 1 (CSLAdV-1). CSLAdV-1 causes viral hepatitis in CSL, and has recently been reported in different species of otariids in an aquarium in Japan ( Otaria flavescens and Arctocephalus pusillus ) and a sequence from Spain has been submitted in NCBI as Otaria flavescens adenovirus-1. The polyomavirus in this animal is a novel virus, and is the first polyomavirus discovered in Hawaiian monk seals. This new virus is designated Hawaiian monk seal polyomavirus (HMSPyV-1), and is 83% homologous to California sea lion Polyomavirus-1 (CSLPyV-1). This is the first report of viral coinfection in a HMS and clinical significance in this case remains unclear but may be associated with advanced age.

Increased mortality and AIDS-like immunopathology in wild chimpanzees infected with SIVcpz.

African primates are naturally infected with over 40 different simian immunodeficiency viruses (SIVs), two of which have crossed the species barrier and generated human immunodeficiency virus types 1 and 2 (HIV-1 and HIV-2). Unlike the human viruses, however, SIVs do not generally cause acquired immunodeficiency syndrome (AIDS) in their natural hosts. Here we show that SIVcpz, the immediate precursor of HIV-1, is pathogenic in free-ranging chimpanzees. By following 94 members of two habituated chimpanzee communities in Gombe National Park, Tanzania, for over 9 years, we found a 10- to 16-fold higher age-corrected death hazard for SIVcpz-infected (n = 17) compared to uninfected (n = 77) chimpanzees. We also found that SIVcpz-infected females were less likely to give birth and had a higher infant mortality rate than uninfected females. Immunohistochemistry and in situ hybridization of post-mortem spleen and lymph node samples from three infected and two uninfected chimpanzees revealed significant CD4(+) T-cell depletion in all infected individuals, with evidence of high viral replication and extensive follicular dendritic cell virus trapping in one of them. One female, who died within 3 years of acquiring SIVcpz, had histopathological findings consistent with end-stage AIDS. These results indicate that SIVcpz, like HIV-1, is associated with progressive CD4(+) T-cell loss, lymphatic tissue destruction and premature death. These findings challenge the prevailing view that all natural SIV infections are non-pathogenic and suggest that SIVcpz has a substantial negative impact on the health, reproduction and lifespan of chimpanzees in the wild.

RETROSPECTIVE REVIEW OF THE PREVALENCE OF MYELOLIPOMA IN GOELDI'S MONKEYS (CALLIMICO GOELDII).

Myelolipomas are mesenchymal tumors composed of adipose and hematopoietic tissue. They have been reported in many species, including domestic dogs and cats, birds, mustelids, wild felids, and nonhuman primates. Myelolipomas in Callitrichidae have been reported at postmortem examination and rarely antemortem. Multiple cases of hepatic myelolipomas associated with morbidity in Chicago Zoological Society's collection of Goeldi's monkeys (Callimico goeldii) prompted a global retrospective study to determine the prevalence and investigate factors associated with this condition. A total of 842 postmortem examination reports (1965-2013) collected from 133 captive collections were reviewed. Myelolipomas were reported in 17.2% (n=145) of animals, with significantly more female cases than male (P=0.023). There was a significantly older mean age at death in affected animals (134 months) compared to unaffected animals (79 months) (P<0.0001). Myelolipomas were diagnosed in the liver in all affected animals, and rarely in the adrenal or spleen. One in five (22.1%) affected Callimico had notable morbidity associated with this disease, which is markedly different from the primarily benign reports in other species. To identify disease, prevent morbidity from hepatic myelolipoma, and improve care of Callimico species in zoological collections, new Species Survival Plan (SSP) medical management recommendations include incorporating abdominal ultrasonographic examination during routine physical examinations in addition to previously recommended hematological and serum biochemical evaluations. Future studies are merited to examine the prevalence, risk factors, health effects, and treatment options of myelolipoma in living collections.

REPRODUCTIVE NEOPLASMS IN WILD AND LONG-TERM CAPTIVE FEMALE FLORIDA MANATEES (TRICHECHUS MANATUS LATIROSTRIS).

Few reports of neoplastic diseases in manatees exist in the veterinary literature. This case series presents reproductive neoplasia noted in eight wild and long-term captive female Florida manatees (Trichechus manatus latirostris) obtained through carcass recovery and animal rehabilitation programs between April 2009 and May 2014. All cases were evaluated histologically, and diagnoses of uterine carcinoma (n = 1), granulosa cell tumor (n = 2), ovarian adnexal tumor (n = 1), and leiomyoma (n = 5) were made. The underlying cause of tumor development and effects on reproductive success is currently unknown, but possible asymmetric reproductive aging and/or a correlation between obesity and reproductive disorder in long-term nonreproductive female manatees are of interest and warrant further investigation.

Human metapneumovirus infection in chimpanzees, United States.

Zoonotic disease transmission and infections are of particular concern for humans and closely related great apes. In 2009, an outbreak of human metapneumovirus infection was associated with the death of a captive chimpanzee in Chicago, Illinois, USA. Biosecurity and surveillance for this virus in captive great ape populations should be considered.

Parasitism and branchitis in various fish species from 4 Cook County, Illinois inland freshwater lakes, 5-year survey, 2017-2021.

We investigated disease trends of concern for fish or public health in a 5-y (2017-2021) prospective survey of fish in Cook County, IL, inland lakes. Lesions were assessed in relation to fish species, lake type and location, season and collection year, and lake water chemistry parameters. Fish included bullheads (n = 98), common carp (n = 99), bluegill (n = 114), and largemouth bass (n = 118). Annually, fish collection and point-source water sampling were conducted in spring, summer, and fall from both seepage and impoundment lakes. Examinations included autopsy, wet-mount cytologic assessment for ectoparasites, and histopathology. No lesions of public health concern were detected. The most common abnormalities were branchitis (261 of 429; 60.8%) and endoparasitism (312 of 429; 72.7%). Branchitis was mild in most cases (189 of 261; 72.4%) and concurrent with branchial parasitism in 175 of 261 (67%) cases. Monogeneans were the most common gill parasites but did not influence branchitis severity (Kruskal-Wallis, p = 0.484). Using generalized ordered logistic regression, predictors of branchitis severity included fish species (p < 0.001), the interaction of lake or location and alkalinity (p < 0.001), and water temperature or season (p < 0.001). Endoparasites included tissue larval trematodes (metacercaria), nematodes, and cestodes (plerocercoids), enteric acanthocephalans, gastric trematodes, renal myxosporidia, biliary and gall bladder myxosporidia, enteric cestodes, and tissue microsporidia. Using generalized ordered logistic regression, variables influencing endoparasitism severity included species (p < 0.001), year (p < 0.001), chloride level (p = 0.009), and the interaction of year and chloride level (p < 0.001). Our results suggested overall good health of fish in sampled Cook County inland lakes and provide a foundation for continued monitoring of ecosystem and public health in the urban environment.

Brevetoxin in blood, biological fluids, and tissues of sea turtles naturally exposed to Karenia brevis blooms in central west Florida.

In 2005 and 2006, the central west Florida coast experienced two intense Karenia brevis red tide events lasting from February 2005 through December 2005 and August 2006 through December 2006. Strandings of sea turtles were increased in the study area with 318 turtles (n = 174, 2005; n = 144, 2006) stranding between 1 January 2005 and 31 December 2006 compared to the 12-yr average of 43 +/- 23 turtles. Live turtles (n = 61) admitted for rehabilitation showed clinical signs including unresponsiveness, paresis, and circling. Testing of biological fluids and tissues for the presence of brevetoxin activity by enzyme-linked immunosorbent assay found toxin present in 93% (52 of 56) of live stranded sea turtles, and 98% (42 of 43) of dead stranded sea turtles tested. Serial plasma samples were taken from several live sea turtles during rehabilitation and toxin was cleared from the blood within 5-80 days postadmit depending upon the species tested. Among dead animals the highest brevetoxin levels were found in feces, stomach contents, and liver. The lack of significant pathological findings in the majority of animals necropsied supports toxin-related mortality.

Disseminated Histiocytic Sarcoma in Two Free-Living Florida Manatees (Trichechus manatus latirostris).

During necropsy of two manatees, masses in multiple organs including liver and spleen were identified. They were composed of neoplastic round to spindle-shaped cells, positive for vimentin, Iba-1, lysozyme, and Mac387, consistent with histiocytic sarcoma. One manatee also had an undifferentiated sarcoma that may have arisen from the histiocytic sarcoma.

Pathologic impacts of contaminants in freshwater fish of Cook County IL.

Mercury (Hg) and polychlorinated biphenyls (PCBs) are widespread environmental toxicants in urban environments with negative impacts to fish health. The present study evaluated the potential association between muscle tissue contaminant (total Hg and total PCB) concentrations and indicators of health in benthic and predatory fish collected from four Forest Preserves of Cook County lakes in the Chicago metropolitan area. Common carp (carp; Cyprinus carpio) and largemouth bass (LMB; Micropterus salmoides) were sampled three times a year (spring, summer, fall) during 2019 and 2020. Water quality analyses (temperature, color, turbidity, dissolved oxygen, total alkalinity, pH, chloride, nitrate, phosphate, ammonium, and pH) were performed concurrently with fish collections. Tissue (skin-on fillet) contaminant concentrations were compared between lake types and fish species and assessed for any relationship with fish morphometric data and pathologic lesions. Main health indicator endpoints included muscle lipid content, parasite burden, and pathologic lesions. Mean total PCB concentrations were greater in carp (203.1 ± 152 µg/kg, wet weight), and mean Hg concentrations were greater in LMB (0.11 ± 0.1 mg/kg, wet weight). In most fish, concentrations of both toxicants surpassed the EPA's lowest threshold to restrict fish consumption for sensitive cohorts (0.029 mg/kg for Hg and 1.5 µg/kg for PCBs). In both species, Hg positively correlated with splenic pigmented macrophage aggregate area (P < 0.001). In carp, Hg also positively correlated with hepatocellular pigmentation (P < 0.01). Mercury correlated with standard length in both species (LMB: P < 0.001, carp: P = 0.95), but polychlorinated biphenyls only correlated with standard length in carp (P < 0.001). No association was found between intraspecific contaminant concentrations and parasite burden, year, or lake type, though differences were noted among individual lakes. The contaminant burden appeared well-tolerated with only mild Hg-associated and no appreciable PCB-associated lesions. However, possible effects on reproduction or behavior were not fully assessed, and future studies are warranted.