The Study of Bluetongue Virus (BTV) and Epizootic Hemorrhagic Disease Virus (EHDV) Circulation and Vectors at the Municipal Parks and Zoobotanical Foundation of Belo Horizonte, Minas Gerais, Brazil (FPMZB-BH).

Bluetongue Virus (BTV) and Epizootic Hemorrhagic Disease Virus (EHDV) are Orbiviruses primarily transmitted by their biological vector, Culicoides spp. Latreille, 1809 (Diptera: Ceratopogonidae). These viruses can infect a diverse range of vertebrate hosts, leading to disease outbreaks in domestic and wild ruminants worldwide. This study, conducted at the Belo Horizonte Municipal Parks and Zoobotany Foundation (FPMZB-BH), Minas Gerais, Brazil, focused on Orbivirus and its vectors. Collections of Culicoides spp. were carried out at the FPMZB-BH from 9 December 2021 to 18 November 2022. A higher prevalence of these insects was observed during the summer months, especially in February. Factors such as elevated temperatures, high humidity, fecal accumulation, and proximity to large animals, like camels and elephants, were associated with increased Culicoides capture. Among the identified Culicoides spp. species, Culicoides insignis Lutz, 1913, constituted 75%, and Culicoides pusillus Lutz, 1913, 6% of the collected midges, both described as competent vectors for Orbivirus transmission. Additionally, a previously unreported species in Minas Gerais, Culicoides debilipalpis Lutz, 1913, was identified, also suspected of being a transmitter of these Orbiviruses. The feeding preferences of some Culicoides species were analyzed, revealing that C. insignis feeds on deer, Red deer (Cervus elaphus) and European fallow deer (Dama dama). Different Culicoides spp. were also identified feeding on humans, raising concerns about the potential transmission of arboviruses at the site. In parallel, 72 serum samples from 14 susceptible species, including various Cervids, collected between 2012 and 2022 from the FPMZB-BH serum bank, underwent Agar Gel Immunodiffusion (AGID) testing for BTV and EHDV. The results showed 75% seropositivity for BTV and 19% for EHDV. Post-testing analysis revealed variations in antibody presence against BTV in a tapir and a fallow deer and against EHDV in a gemsbok across different years. These studies confirm the presence of BTV and EHDV vectors, along with potential virus circulation in the zoo. Consequently, implementing control measures is essential to prevent susceptible species from becoming infected and developing clinical diseases.

ANTIMICROBIAL SUSCEPTIBILITY OF STAPHYLOCOCCUS SPP. ISOLATED FROM FELIDS AND CANIDS IN BELO HORIZONTE ZOO, BRAZIL.

The epidemiology of Staphylococcus spp. has become a major concern among humans and animals due to increasing antimicrobial resistance and frequent reports of infection. Despite the importance of animals as reservoirs for staphylococci, little is known about the epidemiology of Staphylococcus spp. in most nondomestic species, including canids and felids. This study evaluated the frequency, distribution, and patterns of antimicrobial resistance of staphylococcal species isolated from captive felids and canids from Belo Horizonte Zoo, Brazil. Rectal, oral, and nasal swabs from apparently healthy maned wolves (Chrysocyon brachyurus, n= 7), a lion (Panthera leo, n = 1), jaguars (Panthera onca, n = 3), and one swab of a cougar (Puma concolor) with an ear infection were streaked onto mannitol salt agar. Colonies identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry, polymerase chain reaction for the Staphylococcus intermedius group (SIG), and 16S rRNA gene sequencing. Isolates were subjected to antimicrobial susceptibility tests and Staphylococcus pseudintermedius strains were subjected to multilocus sequence typing. Staphylococcus species were isolated from 24 of the 34 samples (70.6%). Among the isolated strains, S. pseudintermedius and Staphylococcus felis were the most frequent species (41.7 and 25%, respectively). Five novel sequence types were identified among the S. pseudintermedius isolates. Resistance to tetracycline (7/24, 29.2%) or penicillin (6/23, 26.1%) was significantly higher than the other antimicrobial agents tested (P < 0.05). One isolate, Staphylococcus nepalensis, was positive for mecA and resistant to five antimicrobials, and was thus classified as multidrug-resistant. The present work suggests that maned wolves are natural hosts of SIG and also reports the isolation of S. felis in sick and healthy, captive, nondomestic carnivores. The isolated staphylococci were susceptible to most classes of antimicrobials tested. However, the multidrug-resistance capability of an S. nepalensis strain reinforces the hypothesis that felids and canids act as reservoirs of pathogens with antimicrobial resistance.

Chylothorax associated with pulmonary compressive atelectasis in an emperor tamarin (Saguinus imperator).

Chylothorax is the accumulation of lymph in the thoracic cavity, and it has never been reported in neotropical primates. An emperor tamarin died and at necropsy chylothorax associated with pulmonary compressive atelectasis was diagnosed. Idiopathic chylothorax can be a cause of respiratory insufficiency and death in tamarins.

Polyarteritis nodosa in a captive common wooly monkey (Lagothrix lagotricha) associated with intestinal necrosis, peritonitis, and sepsis.

Polyarteritis nodosa is an idiopathic necrotizing vasculitis that affects small to medium-sized arteries. We describe a case of polyarteritis nodosa in a captive common wooly monkey (Lagothrix lagotricha) associated with transmural intestinal necrosis and secondary peritonitis. This condition must be considered for differential diagnosis of segmental arteritis in neotropical primates.

Lethal acute diarrhea associated with Clostridioides difficile toxin A and B in a buffy-tufted-ear marmoset (Callithrix aurita).

This is a case of lethal acute diarrhea associated with a mild neutrophilic enteritis in a buffy-tufted-ear marmoset (Callithrix aurita) with detection of A/B toxins and isolation of a toxigenic clade 3 Clostridioides difficile strain (A+ B+ CDT+ , ST5), which should be considered as a potential cause of enteritis in this species.

A RETROSPECTIVE STUDY OF HEPATIC HEMOSIDEROSIS AND IRON STORAGE DISEASE IN SEVERAL CAPTIVE AND FREE-RANGING AVIAN SPECIES.

Some wild species of mammals and birds are prone to excessive iron accumulation, especially when maintained in human care. Hemosiderosis is the process of intracellular accumulation of iron without evidence of toxicity, whereas hemochromatosis is characterized by severe iron accumulation with accompanying organ damage. Iron storage disease (ISD) occurs when organ damage is severe and causing clinical signs. This retrospective study investigated the occurrence of hemosiderosis and ISD across a variety of avian taxa, including captive and free-ranging birds. Archived paraffin-embedded hepatic samples from 103 birds from Belo Horizonte Zoo that died naturally in the period of 2008 to 2018 were re-evaluated with histologic and morphometric techniques, focusing on the identification and scoring of iron deposits in hepatocytes and the quantification of total affected hepatic area. The birds represented 13 orders, 22 families, and 52 genera, and 66 (64.0%) had some degree of iron accumulation in their liver. Importantly, no statistical difference was observed in the occurrence of iron accumulation between families, orders, or origin (free-ranging or captive). Direct and positive correlation was observed between the total area affected by the iron deposits and the histologic score. In this study, there were two cases with severe iron accumulation and clinical signs compatible with ISD: a barefaced curassow (Crax fasciolata) and a channel-billed toucan (Ramphastos vitellinus). This study indicates that iron accumulation may occur in a wide range of avian species, with frequencies and intensities that are similar between free-ranging birds and those in human care. It describes for the first time the occurrence of ISD in a Galliform species.

Mammaliicoccus (Staphylococcus) sciuri-induced suppurative meningoencephalitis and bacteremia in an infant western lowland gorilla (Gorilla gorilla gorilla).

Mammaliicoccus (Staphylococcus) sciuri has been rarely associated with infections and sepsis in humans. A 3-month-old male western lowland gorilla (Gorilla gorilla gorilla), born under human care, died after a traumatic event. Histologic, microbiologic, and molecular findings in postmortem demonstrated a suppurative meningoencephalitis and bacteremia associated with M. sciuri infection.

Streptococcus pasteurianus-induced valvular endocarditis and sepsis in a puerperal emperor tamarin (Saguinus imperator).

Streptococcus pasteurianus is associated with endocarditis and sepsis in humans. A puerperal emperor tamarin died, and necropsy showed a bacterial endocarditis with sepsis. DNA sequencing from the paraffinized heart tissue was compatible with S. pasteurianus. S. pasteurianus could be an important agent associated with sepsis in tamarins.

Natural cururu toad (Rhinella sp.) poisoning in a free-ranging lesser grison (Galictis cuja): Outcomes in a new susceptible predator with a novel peptide description.

Cururu toad (Rhinella marina group) is widely distributed in Brazil. Lesser grison (Galitic cuja) is a South American mustelid. This is the first report of natural poisoning in a free-ranging lesser grison by Rhinella toad parotoid gland secretion (PGS). Five minutes after biting a toad, the lesser grison developed convulsion, dying within 1.5 h. Mass spectrometry analysis of a milky-whitish secretion found in the lesser grison oral cavity allowed identification of a bufotoxin and a new bufonid peptide.

ACUTE NECROTIZING AND EOSINOPHILIC MYOCARDITIS IN A CHIMPANZEE (PAN TROGLODYTES).

Cardiac disease is of importance in captive chimpanzee (Pan troglodytes) health. Here we report an eosinophilic and necrotizing myocarditis in a 17-y-old chimpanzee with no previous history of cardiac disease that progressed to death within 48 h. Toxic and infectious causes were ruled out. The chimpanzee had eosinophilia at different occasions in previous years. The animal had a severe, diffuse, and acute monophasic necrotizing myocarditis, with a moderate lymphoplasmacytic infiltrate that was rich in eosinophils. Ante- and postmortem investigations are compatible with an unusual eosinophilic myocarditis with clinical evolution and morphology comparable with human eosinophilic myocarditis secondary to hypereosinophilic syndrome.

Polyostotic osteosarcoma associated with avian leukosis virus infection in a captive bare-faced curassow (Crax fasciolata).

BACKGROUND: Osteosarcoma is a malignant mesenchymal bone tumor. Although it is a common tumor in the appendicular skeleton of dogs and cats, it is rarely reported in birds. Retroviruses are usually associated with solid tumor development in different avian species. CASE PRESENTATION: This report aims to describe a case of osteosarcoma associated with the avian leukosis virus in a captive bare-faced curassow (Crax fasciolata). A captive adult female bare-faced curassow presented with lameness, hyporexia, and a non-ulcerative and firm tumor in the right femur. The bird was euthanized due to the poor prognosis. Histopathology revealed an infiltrative mesenchymal neoplasm consisting of spindle cells with moderate cell pleomorphism, organized in bundles and interspersed by marked deposition of the osteoid matrix, which was compatible with osteosarcoma affecting both femur and tibiotarsus, with renal metastasis. Immunohistochemistry of the primary and metastatic tumor demonstrated vimentin expression by neoplastic cells. Samples of the neoplasm, bone marrow, and spleen were processed for PCR, which enabled the demonstration of proviral avian leukosis virus (ALV) DNA. CONCLUSIONS: To the best of our knowledge, this is the first report of an osteosarcoma in a bare-faced curassow with an unusual polyostotic manifestation and associated with ALV infection.

Genital infection by Aerococcus viridans in a captive african elephant (Loxodonta africana) / Infecção genital por Aerococcus viridans em um elefante africano em cativeiro (Loxodonta africana)

ABSTRACT: Aerococcus viridans is an emerging pathogen for humans and livestock animals, mainly associated with genitourinary infections cases. Its occurrence in wild mammals has never been reported. The aim of this study was to determine the etiological agent associated with clinical a case of a genital infection in a female African elephant (Loxodonta africana). Phylogenetic analysis and antimicrobial susceptibility profile of the isolate were also addressed. The animal presented frequent cases of genital infection with intermittent white secretion. Purulent secretion was sampled and submitted to bacteriological exam. The isolate obtained was thus identified by phenotypic and molecular methods as A. viridans and was found to be similar to human pathogenic isolates in BLASTn and phylogenetic analysis. The isolate was sensitive to almost all antimicrobials evaluated, presenting resistance to ciprofloxacin and norfloxacin. This is the first report of occurrence of A. viridans infection in the genital tract of an African elephant.

RESUMO: Aerococcus viridans é um patógeno emergente para seres humanos e animais de produção, principalmente associado a casos de infecções geniturinárias. Sua ocorrência em mamíferos selvagens nunca foi relatada. O objetivo deste estudo foi determinar o agente etiológico associado a um caso clínico de infecção genital em uma fêmea de elefante africano (Loxodonta africana). Análises filogenéticas e perfil de susceptibilidade antimicrobiana do isolado também foram avaliados. O animal apresentou casos frequentes de infecção genital com eliminação de secreção branca intermitente. A secreção purulenta foi coletada e submetida a exame bacteriológico. O isolado obtido foi identificado por métodos fenotípicos e moleculares como A. viridans e apresentou alta similaridade a isolados humanos patogênicos nas análises de BLASTn e filogenética. O isolado foi sensível a quase todos os antimicrobianos avaliados, apresentando resistência à ciprofloxacina e norfloxacina. Este é o primeiro relato de ocorrência de infecção por A. viridans no trato genital de elefante africano.

PATHOLOGIC FINDINGS IN 36 SLOTHS FROM BRAZIL.

Sloths are xenarthrans from Central and South America with a highly adapted morphophysiology. Five of the six known species of sloths are found in Brazil, among which Bradypus torquatus (maned three-toed sloth) is considered a vulnerable species by International Union for Conservation of Nature. Nevertheless, knowledge on health and disease of sloths is very scarce, thus this study aimed to describe macroscopic and microscopic findings in 36 Brazilian sloths. The most common findings included iron storage disorder, probable bacterial pneumonia, gastric and intestinal nematode parasitism, and a presumptive diagnosis of systemic mastocytosis.

Serological evidence of Leishmania infection by employing ELISA and rapid tests in captive felids and canids in Brazil.

Visceral leishmaniasis (VL) is a zoonosis and the dog is considered the most important urban reservoir. Cases in cats have been reported, but little is known about Leishmania infection and disease in wild felids and canids kept in captivity in endemic areas. Thus, the serological pattern of wild felids and canids kept in captivity at the Belo Horizonte Zoological Garden was investigated using two primary antigens for conventional ELISA: k39 and rKDDR, as well as three serological rapid kits: Dual Path Platform (DPP®) immunochromatographic test, rKDDR Immunochromatographic assay and ELISA SNAP Leishmania IDEXX®. A total of 21 serum samples, 13 of wild felids and 8 wild canids of varying age and sex were evaluated. The results obtained in the tests were analyzed by agreement using Kappa coefficient, and between ELISA antigens all the analysis performed had showed significant agreement among both of them, as well between the three immunochromatographic tests. The results demonstrated that there is serological evidence of wild animals seropositive for Leishmania antibodies at the Belo Horizonte Zoological Garden, and that all the antigens and rapid tests used can be employed in serological screening for VL in wild felids and canids.

Competence of non-human primates to transmit Leishmania infantum to the invertebrate vector Lutzomyia longipalpis.

Leishmaniasis is a zoonotic disease of worldwide relevance. Visceral leishmaniasis is endemic in Brazil, where it is caused by Leishmania infantum with Lutzomyia longipalpis being the most important invertebrate vector. Non-human primates are susceptible to L. infantum infection. However, little is known about the role of these species as reservoirs. The aim of this study was to evaluate the transmissibility potential of visceral leishmaniasis by non-human primates through xenodiagnosis using the phlebotomine Lu. longipalpis as well as to identify phlebotomine species prevalent in the area where the primates were kept in captivity, and assess infection by Leishmania in captured phlebotomine specimens. Fifty two non-human primates kept in captivity in an endemic area for leishmaniasis were subjected to xenodiagnosis. All primates were serologically tested for detection of anti-Leishmania antibodies. Additionally, an anti-Lu. longipalpis saliva ELISA was performed. Sand flies fed on all animals were tested by qPCR to identify and quantify L. infantum promastigotes. Eight of the 52 non-human primates were positive by xenodiagnosis, including three Pan troglodytes, three Leontopithecus rosalia, one Sapajus apella, and one Miopithecus talapoin, with estimated numbers of promastigotes ranging from 5.67 to 1,181.93 per µg of DNA. Positive animals had higher levels of IgG anti-Lu. longipalpis saliva when compared to negative animals, prior to xenodiagnosis. Captive non-human primates are capable of infecting Lu. longipalpis with L. infantum. Our findings also demonstrate the relevance of non-human primates as sentinels to zoonotic diseases. Several phlebotomine species, including Lu. longipalpis, have been identified in the area where the primates were maintained, but only one pool of Lutzomyia lenti was infected with L. infantum. This study has implications for public health strategies and conservation medicine.

Visceral leishmaniasis in an infant gorilla (Gorilla gorilla gorilla): Clinical signs, diagnosis, and successful treatment with single-dose liposomal amphotericin B.

A 2-year-old captive gorilla (Gorilla gorilla gorilla) was diagnosed with visceral leishmaniasis in Brazil, and treated with a single dose of liposomal amphotericin B, which resulted in clinical cure. This is the first report of visceral leishmaniasis in gorillas, and the first reported liposomal amphotericin B treatment in great apes.

Environmental Enrichment Effect on Fecal Glucocorticoid Metabolites and Captive Maned Wolf (Chrysocyon brachyurus) Behavior.

Environmental enrichment is a technique that may reduce the stress of nonhuman animals in captivity. Stress may interfere with normal behavioral expression and affect cognitive decision making. Noninvasive hormonal studies can provide important information about the stress statuses of animals. This study evaluated the effectiveness of different environmental enrichment treatments in the diminution of fecal glucocorticoid metabolites (stress indicators) of three captive maned wolves (Chrysocyon brachyurus). Correlations of the fecal glucocorticoid metabolite levels with expressed behaviors were also determined. Results showed that environmental enrichment reduced fecal glucocorticoid metabolite levels. Furthermore, interspecific and foraging enrichment items were most effective in reducing stress in two of the three wolves. No definite pattern was found between behavioral and physiological responses to stress. In conclusion, these behavioral and physiological data showed that maned wolves responded positively from an animal well being perspective to the enrichment items presented.

Pathogens of wild maned wolves (Chrysocyon brachyurus) in Brazil.

The maned wolf, Chrysocyon brachyurus, is an endangered Neotropical canid that survives at low population densities. Diseases are a potential threat for its conservation but to date have been poorly studied. We performed clinical evaluations and investigated the presence of infectious diseases through serology and coprologic tests on maned wolves from Galheiro Natural Private Reserve, Perdizes City, Minas Gerais State, southeastern Brazil. Fifteen wolves were captured between 2003 and 2008. We found high prevalences of antibody to canine distemper virus (CDV; 13/14), canine parvovirus (CPV; 4/14), canine adenovirus type 2 (13/14), canine coronavirus (5/11), canine parainfluenza virus (5/5), and Toxoplasma gondii (6/8), along with Ancylostomidae eggs in all feces samples. Antibodies against Leishmania sp. were found in one of 10 maned wolves, and all samples were negative for Neospora caninum. Evidence of high exposure to these viral agents was also observed in unvaccinated domestic dogs from neighboring farms. High prevalence of viral agents and parasites such as CDV, CPV, and Ancylostomidae indicates that this population faces considerable risk of outbreaks and chronic debilitating parasites. This is the first report of exposure to canine parainfluenza virus in Neotropical free-ranging wild canids. Our findings highlight that canine pathogens pose a serious hazard to the viability of maned wolves and other wild carnivore populations in the area and emphasize the need for monitoring and protecting wildlife health in remaining fragments of the Cerrado biome.

Behavioral responses of maned wolves (Chrysocyon brachyurus, Canidae) to different categories of environmental enrichment stimuli and their implications for successful reintroduction.

The maned wolf (Chrysocyon brachyurus, Illiger, 1815, Canidae) is a threatened species that inhabits the cerrados of Brazil, Argentina, Peru, Bolivia, Paraguay, and Uruguay. Captive maned wolves could be potentially used in reintroduction programs for species conservation; however, it is necessary that their behavior and cognitive abilities are conserved. Environmental enrichment is a tool used to stimulate captive animals and maintain a natural behavioral repertoire. To compare the variation of captive maned wolves' behavioral responses to environmental enrichment, we studied three maned wolves held by Belo Horizonte Zoo, Brazil. Foraging, interspecific, and intraspecific stimuli were offered to the animals and their responses were compared with a baseline and postenrichment treatments. The test was used to help in choosing which one of the three individuals would participate in a reintroduction project. The results showed that stimuli type did influence the animal's responses, and that individually wolves responded differently to foraging, interspecific, and intraspecific enrichment items (P80.05 for some behaviors in each enrichment category). The individual's personality seemed to influence their behavioral responses, with animals showing bold and shy responses, and this trait should be considered during decision makings for reintroduction.

First observations of nest attendance behavior by wild maned wolves, Chrysocyon brachyurus.

In many species of canids the male's role in reproduction extends to providing extensive or some parental care to his offspring. Maned wolves are a monogamous canid species whose males have been observed providing parental care to their offspring in captivity, but no field observations exist. We observed a wild pair of maned wolves at their nest site in a period soon after the female had given birth for a total period of 65 days. We made five observation sessions with an average of 3-4 days each separated by approximately 2 weeks. Direct visual observations of maned wolves were made each 30 min during the hours of darkness (17:00-07:00) using night-vision binoculars and confirmed by VHF radiotelemetry. During observations we recorded the location of the male and the female in relation to the nest (i.e., in the nest, nearby or long way from the nest). The results showed that the female spent more than 60.44% of her time in or near the nest. The male spent 28.90% of his time in or near the nest. There was a positive significant correlation between the female and the male in terms of the amount of time spent in or near the nest (P<0.01). The maned wolves showed a strong temporal variation in time spent in or near the nest. In conclusion, our data show that wild male maned wolves--probably--provide parental care through provision of food to their female and presumed offspring rather than "babysitting."