Lack of serological and molecular evidences of Zika virus circulation in non-human primates in three states from Brazil.

BACKGROUND: Zika virus (ZIKV) was discovered in 1947 with the virus isolation from Rhesus monkey (Macaca mulatta) in Uganda forest, Africa. Old World Primates are involved in a sylvatic cycle of maintenance of this arbovirus, however a limited knowledge about the role of New World primates in ZIKV transmission cycles has been established. OBJECTIVE: This work aimed to investigate the presence of enzootic circulation of ZIKV in New World Primates from three Brazilian states: São Paulo, Paraíba, and Paraná. METHODS: We analyzed 100 non-human primate samples collected in 2018 and 2020 from free-ranging and captive environments from São Paulo (six municipalities belonging to Sorocaba region), Paraíba (João Pessoa municipality), and Paraná (Foz do Iguaçu municipality) using reverse transcriptase quantitative polymerase reaction (RT-qPCR) assays, indirect enzyme-linked immunosorbent assay (ELISA), and plaque reduction neutralization test (PRNT). FINDINGS: All samples (n = 141) tested negative for the presence of ZIKV genome from tissue and blood samples. In addition, all sera (n = 58) from Foz do Iguaçu' non-human primates (NHPs) were negative in serological assays. MAIN CONCLUSION: No evidence of ZIKV circulation (molecular and serological) was found in neotropical primates. In addition, the absence of antibodies against ZIKV suggests the absence of previous viral exposure of NHPs from Foz do Iguaçu-PR.

A pilot study about assisted reproduction in harpy eagles (Harpia harpyja) in the course of species conservation including collection, storage, and analysis of semen.

Currently, 52% of all raptor species demonstrate a decreasing population tendency, and the American harpy eagle (Harpia harpyja) has been categorized as "near threatened" by the IUCN. Habitat loss, persecution, and subsequent reduction of genetic diversity are regarded as major threats to the world's strongest eagle. Captive breeding and reintroduction into protected habitats are approaches of species conservation projects, but captive propagation is difficult due to low ex-situ numbers and scarce successful breeding pairs. The aim of the present study was to collect, analyze, and store semen from harpy eagles and to use aliquots for artificial insemination to increase the number of offspring and to include more individuals into the ex-situ gene pool. First, semen collection and semen availability were assessed in four males during the course of 1 year in European zoos. Second, these experiences were transferred to ex-situ breeding programs in Brazilian zoos to attempt semen collection in 13 male eagles. Semen collection was successful in 51.7% of the attempts and in 8/13 males (individual success rates 20-100%) using electro-stimulation. Most commonly, whey-like to milky, whitish semen samples were collected, regularly containing urate impurities (67.7%). The median semen volume was 106 µl and the median sperm concentration 5,000 sperm/µl (750-22,500 sperm/µl). Mean values for pH were 6.7, for sperm motility 27.7 ± 22.6%, for progressive motility 2.9 ± 5.6%, and for sperm viability 46.6 ± 16.3%. Using semen extenders, a sperm motility of 8% was maintained for 27 h in the refrigerator. Artificial insemination was performed in one female, but the success of fertilization could not be assessed due to egg destruction. In this study, methods for assisted reproduction were refined for use in harpy eagles, and the first semen samples were evaluated as a start to establish species-specific orientation values.

Canine distemper virus and canine adenovirus type-2 infections in neotropical otters (Lontra longicaudis) from Southern Brazil.

All descriptions of infectious diseases affecting otters were published in the Northern Hemisphere, with no occurrence identified in neotropical otters (Lontra longicaudis). Consequently, a retrospective histopathological study using archival tissue samples from six free-living neotropical otters was done to investigate the possible occurrence of disease patterns associated with common viral infectious disease agents of the domestic dogs. Immunohistochemical (IHC) assays were designed to identify intralesional tissue antigens of canine distemper virus (CDV), and canine adenovirus-1 (CAdV-1) and canine adenovirus-2 (CAdV-2). The most frequent histopathological patterns diagnosed were interstitial pneumonia (83.33%; 6/5) and hepatocellular vacuolar degeneration (50%; 3/6). IHC identified intralesional intracytoplasmic immunoreactivity to CDV antigens in all otters evaluated, with positive immunolabeling occurring within epithelial cells of the lungs, stomach, kidneys, and liver, and skin. Intracytoplasmic CAdV-2 antigens were identified within epithelial cells of the peribronchial glands in four otters with interstitial pneumonia. These findings resulted in singular and simultaneous infections in these neotropical otters, represented the first report of concomitant infections by CDV and CAdV-2 in free-living neotropical otters from the Southern Hemisphere, and suggested that this mammalian species is susceptible to infections by viral disease agents common to the domestic dogs and may develop similar histopathologic disease patterns.

Lack of serological and molecular evidences of Zika virus circulation in non-human primates in three states from Brazil

BACKGROUND Zika virus (ZIKV) was discovered in 1947 with the virus isolation from Rhesus monkey (Macaca mulatta) in Uganda forest, Africa. Old World Primates are involved in a sylvatic cycle of maintenance of this arbovirus, however a limited knowledge about the role of New World primates in ZIKV transmission cycles has been established. OBJECTIVE This work aimed to investigate the presence of enzootic circulation of ZIKV in New World Primates from three Brazilian states: São Paulo, Paraíba, and Paraná. METHODS We analyzed 100 non-human primate samples collected in 2018 and 2020 from free-ranging and captive environments from São Paulo (six municipalities belonging to Sorocaba region), Paraíba (João Pessoa municipality), and Paraná (Foz do Iguaçu municipality) using reverse transcriptase quantitative polymerase reaction (RT-qPCR) assays, indirect enzyme-linked immunosorbent assay (ELISA), and plaque reduction neutralization test (PRNT). FINDINGS All samples (n = 141) tested negative for the presence of ZIKV genome from tissue and blood samples. In addition, all sera (n = 58) from Foz do Iguaçu' non-human primates (NHPs) were negative in serological assays. MAIN CONCLUSION No evidence of ZIKV circulation (molecular and serological) was found in neotropical primates. In addition, the absence of antibodies against ZIKV suggests the absence of previous viral exposure of NHPs from Foz do Iguaçu-PR.

'Candidatus Mycoplasma haemoalbiventris' and tick-borne pathogens screening in white-eared opossums (Didelphis albiventris) from Curitiba and Foz do Iguaçu Cities, Paraná State, southern Brazil.

Hemoplasmas are epierythrocytic bacteria that infect mammals. 'Candidatus Mycoplasma haemoalbiventris' was detected in white-eared opossums (Didelphis albiventris) from southern and central-western Brazil. The present study aimed at: i) screening opossums for tick-borne (TBP) pathogens (Piroplasmida and Anaplasmataceae) and ii) detecting and characterizing hemoplasma species infecting opossums from Curitiba and Foz do Iguaçu cities in the Paraná State, southern Brazil. Thirty blood samples from white-eared opossums were evaluated by PCR assays. Animals were not infested by ectoparasites. The mammalian endogenous gapdh gene was consistently amplified in all samples. All opossums tested negative for Theileria/Babesia spp. and Ehrlichia/Anaplasma spp. by PCR based on 18S rRNA and 16S rRNA genes, respectively. A genus-specific PCR assay based on the 16S rRNA gene of hemoplasmas showed that three/13 (23.08%; CI 95%: 8.18-50.26%) opossums from Foz do Iguaçu were positive for hemotropic Mycoplasma sp. All opossums from Curitiba tested negative for hemoplasmas. Sequencing of both the 16S and 23S rRNA genes revealed that the animals were infected by 'Ca. M. haemoalbiventris'. Although 'Ca. M. haemoalbiventris' is prevalent in opossums in Brazil, clinical signs associated with its infection and its putative vectors remain unknown.

'Candidatus Mycoplasma haemoalbiventris' and tick-borne pathogens screening in white-eared opossums (Didelphis albiventris) from Curitiba and Foz do Iguaçu Cities, Paraná State, southern Brazil / 'Candidatus Mycoplasma haemoalbiventris' e triagem para patógenos transmitidos por carrapatos em gambás de orelha branca (Didelphis albiventris) das cidades de Curitiba e Foz do Iguaçu, Estado do Paraná, sul do Brasil

Abstract Hemoplasmas are epierythrocytic bacteria that infect mammals. 'Candidatus Mycoplasma haemoalbiventris' was detected in white-eared opossums (Didelphis albiventris) from southern and central-western Brazil. The present study aimed at: i) screening opossums for tick-borne (TBP) pathogens (Piroplasmida and Anaplasmataceae) and ii) detecting and characterizing hemoplasma species infecting opossums from Curitiba and Foz do Iguaçu cities in the Paraná State, southern Brazil. Thirty blood samples from white-eared opossums were evaluated by PCR assays. Animals were not infested by ectoparasites. The mammalian endogenous gapdh gene was consistently amplified in all samples. All opossums tested negative for Theileria/Babesia spp. and Ehrlichia/Anaplasma spp. by PCR based on 18S rRNA and 16S rRNA genes, respectively. A genus-specific PCR assay based on the 16S rRNA gene of hemoplasmas showed that three/13 (23.08%; CI 95%: 8.18-50.26%) opossums from Foz do Iguaçu were positive for hemotropic Mycoplasma sp. All opossums from Curitiba tested negative for hemoplasmas. Sequencing of both the 16S and 23S rRNA genes revealed that the animals were infected by 'Ca. M. haemoalbiventris'. Although 'Ca. M. haemoalbiventris' is prevalent in opossums in Brazil, clinical signs associated with its infection and its putative vectors remain unknown.

Resumo Hemoplasmas são bactérias epieritrocíticas que infectam mamíferos. 'Candidatus Mycoplasma haemoalbiventris' foi detectado previamente em gambás-de-orelha-branca (Didelphis albiventris) das regiões sul e centro-oeste do Brasil. O presente estudo objetivou: i) triar os gambás para as doenças transmitidas por carrapatos (Piroplasmida e Anaplasmataceae); e ii) detectar e caracterizar as espécies de hemoplasma que infectam gambás nas cidades de Curitiba e Foz do Iguaçu, no Estado do Paraná, sul do Brasil. Trinta amostras de sangue de gambás-de-orelha-branca foram analisadas por PCR. Os animais não estavam infestados por ectoparasitos. O gene endógeno de mamífero gapdh foi amplificado em todas as amostras. Todos os gambás testaram negativos para Theileria/Babesia spp. e Ehrlichia/Anaplasma spp. por PCR, respectivamente, para os genes 18S rRNA e 16S rRNA. Uma PCR gene-específica, baseada no gene 16S rRNA de hemoplasmas, mostrou que três/13 (23,08%; CI 95%: 8,18-50,26%) gambás de Foz do Iguaçu foram positivos para Mycoplasma sp. hemotrópico. Todos os gambás de Curitiba testaram negativos para hemoplasmas. O sequenciamento de fragmentos dos genes 16S e 23S rRNA revelou que os animais estavam infectados pelo 'Ca. M. haemoalbiventris'. Embora 'Ca. M. haemoalbiventris' seja prevalente em gambás no Brasil, os sinais clínicos associados à infecção e os prováveis vetores permanecem desconhecidos.

Immunohistochemical evidence of canine morbillivirus (canine distemper) infection in coatis (Nasua nasua) from Southern Brazil.

The pathological and immunohistochemical (IHC) findings associated with infection due to canine morbilivírus (canine distemper virus, CDV) are described in coatis (Nasua nasua). Tissue fragments of coatis (n = 13) that died at the Bela Vista Sanctuary, Paraná, Southern Brazil, were routinely processed for histopathology to identify the main histopathologic patterns as compared to that of the domestic dog. Selected formalin-fixed paraffin-embedded (FFPE) tissue fragments of the lungs, liver, urinary bladder and small intestine were used in IHC assays designed to identify the antigens of CDV, canine adenovirus (CAdV-1 and CAdV-2) and canine parvovirus type 2 (CPV-2). The main histopathologic patterns identified were interstitial pneumonia (n = 9), interstitial nephritis (n = 6), atrophic enteritis (n = 4) and ballooning degeneration of the uroepithelium (n = 3). Positive immunolabelling for intralesional antigens of CDV was identified in the lung with interstitial pneumonia (n = 3), in the intestine (n = 2) and in the degenerated epithelium of the urinary bladder (n = 2). Antigens of CPV-2, CAdV-1 and CAdV-2 were not identified in any FFPE tissue sections evaluated. These findings indicate that these wild carnivores were infected by a viral disease pathogen common to the domestic dog and develop similar histopathologic findings. Collectively, these findings suggest that these coatis were infected by CDV and can serve as a potential host for this infectious disease pathogen.

Immunohistochemical identification of antigens of canine distemper virus in neotropical felids from Southern Brazil.

The pathologic and immunohistochemical findings associated with infections due to canine distemper virus (CDV) are described in the cougar (Puma concolor), margay (Leopardus wiedii) and jaguarundi (Herpailurus yagouaroundi) from Southern Brazil. Tissue sections of the neotropical felids (n = 3) that died at the Bela Vista Sanctuary, Paraná, Southern Brazil were routinely processed for histopathology to identify possible histopathologic patterns associated with infections due to CDV. Selected formalin-fixed paraffin embedded tissue sections of the lungs and urinary bladder were used in immunohistochemical assays designed to identify the antigens of CDV. The main histopathologic patterns identified were interstitial pneumonia in the margay and jaguarundi, while ballooning degeneration of the transitional epithelium of the urinary bladder was observed in the cougar. Positive immunoreactivity to antigens of CDV was identified within intralesional sections of the lungs of the two wild felids with interstitial pneumonia and in the degenerated urothelium of the cougar. These findings indicate that these neotropical cats were infected by a viral infectious disease pathogen common to the domestic dog and add to the few documented descriptions of CDV-induced infections in wildlife from Brazil.

Cryopreservation of margay (Leopardus wiedii) spermatozoa: Effects of different extenders and frozen protocols.

Sperm cryopreservation offers many benefits to wild felids conservation programs. However, the implementation of these programs is limited by the different responses of each species to the cryopreservation protocols and extenders used, requiring the formulation of species-specific protocols. For this purpose, semen samples from 6 margays (Leopardus wiedii) were submitted to 2 cryopreservation protocols: 1) manual freezing (cooling rate of - 0.33 °C/min at 5 °C/180 min and freezing rate with two steps - 9 °C/min for 2 min and -19.1 °C/min for 2 min) and 2) automatic freezing machine (cooling rate of - 0.25 °C/min at 5 °C/120 min and freezing rate with one step -20 °C/min for 8.3 min) using 2 commercial extenders, an egg yolk-based (Test Yolk Buffer; TYB) and an egg yolk-free extender (AndroMed; MED). Post-thawed sperm quality was assessed at 3 time points (immediately after thawing and 1 and 2 h post-thawed) by sperm motility index (SMI), plasma membrane and acrosomal integrity, and mitochondrial membrane potential (MMP). Regarding SMI, TYB yielded superior results (29.4 ± 3.5%) compared to MED (11.2 ± 2.8%; p < 0.002) immediately after thawing until 2 h after thawing (TYB 3.9 ± 1.7% and MED 0.0 ± 0.0%; p < 0.05). Furthermore, the automated freezing method provided higher motility compared to the manual freezing procedure immediately post-thaw (25.08 ± 3.66% and 15.78 ± 3.29%, respectively) and 1 h post-thaw (13.71 ± 2.56% and 6.03 ± 1.97%, respectively; p < 0.05). The percentage of intact acrosomes and plasma membranes and the percentage of sperm with high MMP were superior for TYB when compared to MED regardless of cryopreservation protocol (p < 0.05). Conversely, the interaction between cryopreservation protocols and extenders was observed for MMP where TYB exhibits better results compared to MED (p < 0.05) in both procedures, but it was higher in automated procedures. For MED, no changes were found in MMP between procedures. Considering only TYB, samples showed higher MMP when submitted to an automated procedure (p < 0.05). In conclusion, the slow cooling rates with shorter time of exposure to glycerol contributed to minimize cryodamage in the Margays' sperm. Moreover, results indicated that association between TYB and automatic freezing machine ensured the minimal quality of spermatozoa after thawing required for further use in in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI).

ARTs in Wild Felid Conservation Programmes in Poland and in the World.

With the exception of the domestic cat, all felid species (Felidae) are currently threatened with extinction in their natural habitat. To develop effective and optimal wild cat conservation programmes with assisted reproductive technology (ART) it is necessary to combine advances from different disciplines of science, starting from the biology of the species, through research into the population and habitat, assisted reproductive technologies, establishment of gene banks, developing bioinformatic systems, and ending with biodiversity and endangered species management. In the last few years knowledge of felid reproduction has expanded considerably thanks to comparative studies utilising the domestic cat as a research model for endangered wild cats. Basic reproductive techniques utilised in both domestic cat breeding and rescuing wild felid populations that are threatened with extinction include semen collection and cryopreservation, artificial insemination, oocyte collection, in vitro maturation, in vitro fertilisation, somatic cloning, and embryo transfer. The main directions in which assisted reproductive technologies are being developed in wild cat conservation implementations and the contribution of Polish research centres in advancing these methods are presented.

Multiple bluetongue virus serotypes causing death in Brazilian dwarf brocket deer (Mazama nana) in Brazil, 2015-2016.

Bela Vista Biological Sanctuary (RBV) is a protected area of Itaipu Binacional, a hydroelectric power company located on the border of Brazil and Paraguay. A captive population of Brazilian dwarf brocket deer (Mazama nana, Cervidae, Artiodactyla) is maintained for conservation purposes. Despite the reproductive success of the animals, outbreaks of a fatal hemorrhagic disease have been registered over the years, compromising conservation efforts. In order to identify the etiological agents of these hemorrhagic diseases, 32 captive Brazilian dwarf brockets were sampled to investigate bluetongue virus (BTV), epizootic hemorrhagic disease (EHD), and adenovirus hemorrhagic disease (AHD), in 2015. Only one deer (1/32; 3.12%) was seropositive for BTV. After this survey, five animals died in the early autumn of 2015 and 2016, again presenting clinical signs of hemorrhagic disease. Using RT-qPCR, RT-PCR and DNA sequencing, five BTV serotypes (3, 14, 18, 19, and 22) were identified in blood and tissues collected during necropsies. These BTV serotypes had not been previously described or isolated in Brazil, either in wild or domestic ruminants. Additionally, differential diagnosis was performed for EHD and AHD, but all samples were negative for both diseases. The multiple distinct BTV serotypes identified in these outbreaks resulted in a high lethality (100%) of Brazilian dwarf brockets and indicated that various BTV serotypes are circulating in the area.

IMMUNOHISTOCHEMICAL AND MOLECULAR EVIDENCE OF PUTATIVE NEORICKETTSIA INFECTION IN COATIS ( NASUA NASUA) FROM SOUTHERN BRAZIL.

The pathologic, molecular, and immunohistochemical findings associated with Neorickettsia helminthoeca are described in coatis ( Nasua nasua). Tissue sections (small intestine, lungs, kidney, liver, and spleen) of coatis ( n = 3) that died at the Bela Vista Biological Refuge, Foz do Iguaçu, Paraná, southern Brazil were routinely processed from histopathology. Selected formalin-fixed paraffin-embedded (FFPE) tissue sections of the small intestine, lungs, and spleen were used in an immunohistochemical (IHC) assay designed to identify the antigens of N. helminthoeca. Additionally, FFPE tissue sections of the small intestine were used to demonstrate antigens of canine parvovirus-2 (CPV-2) by IHC. Histopathology revealed chronic enteritis in all coatis. Parasitic enteritis was diagnosed in two coatis; one of these contained examples of a trematode within the lumen of the small intestine and the ovum of a trematode encysted in the intestinal mucosa. Other significant pathologic findings included interstitial pneumonia ( n = 2) and pyogranulomatous splenitis ( n = 1). Positive immunolabeling for N. helminthoeca was identified within macrophages of the small intestine and reticuloendothelial cells within the germinal centers of the spleen of all coatis; the intestinal trematode was N. helminthoeca IHC-positive. All pulmonary sections revealed negative immunolabeling for N. helminthoeca. Furthermore, the antigens of CPV-2 were not identified in the intestine of any coati. These findings indicate that these coatis were infected by N. helminthoeca, but since clinical and gross pathological findings were not recorded, it is uncertain if this pathogen produced clinical disease in this canid host; therefore, coatis may be asymptomatic or dead-end hosts for this organism.

First molecular screening of Plasmodium species in ungulates from Southern Brazil.

OBJECTIVE: Despite malaria epidemiology has been extensively studied in primates, few studies were conducted in ungulates. After half a century without descriptions of Plasmodium spp. in deer since its first identification, recent research has rediscovered Plasmodium on ungulates in Africa, Asia, North America and South America, including Central Brazil. Here, a captive herd was evaluated in southern Brazil using light microscopy and PCR. DNA samples were tested for fragment amplification of two Plasmodium spp. genes: mitochondrial cytochrome b and small subunit ribosomal RNA. RESULTS: All analyses were negative. However, the tests were performed on samples that were collected at a single time point, and parasitemia may fluctuate over the parasite's life cycle. Thus, the possibility of occult infection cannot be ruled out. Despite the negative results of all of the methods applied, it cannot be categorically stated that these animals are free from Plasmodium sp. infection. Further monitoring and/or multiple sequential sampling may improve the success rate of detecting parasites. Moreover, although this survey of Plasmodium represents the first molecular study on ungulate malaria parasites from Southern Brazil, further analysis of samples from different ungulate species is important for characterizing the epidemiology of Plasmodium of these mammals in this region.

Serosurvey of anti- Leptospira sp. and anti- Toxoplasma gondii antibodies in capybaras and collared and white-lipped peccaries.

INTRODUCTION:: Brazilian native species are reemerging as increasingly free-ranging populations. METHODS:: Sera from 31 capybaras (Hydrochoerus hydrochaeris) and 28 peccaries (Pecari tajacu and Tayassu pecari) were tested for anti-Leptospira and anti-Toxoplasma gondii antibodies using microscopic seroagglutination test. RESULTS:: Nineteen percent of free-ranging and 10.0% of captive capybaras, along with 31.8% of collared peccaries, were seropositive for T. gondii. None was seropositive for Leptospira sp. CONCLUSIONS:: The present findings indicated low risk of disease, particularly among capybaras and white-lipped peccaries; however, active surveillance programs are important for monitoring wildlife health and public health once they are in public parks around cities.

Serosurvey of anti- Leptospira sp. and anti- Toxoplasma gondii antibodies in capybaras and collared and white-lipped peccaries

Abstract INTRODUCTION: Brazilian native species are reemerging as increasingly free-ranging populations. METHODS: Sera from 31 capybaras (Hydrochoerus hydrochaeris) and 28 peccaries (Pecari tajacu and Tayassu pecari) were tested for anti-Leptospira and anti-Toxoplasma gondii antibodies using microscopic seroagglutination test. RESULTS: Nineteen percent of free-ranging and 10.0% of captive capybaras, along with 31.8% of collared peccaries, were seropositive for T. gondii. None was seropositive for Leptospira sp. CONCLUSIONS: The present findings indicated low risk of disease, particularly among capybaras and white-lipped peccaries; however, active surveillance programs are important for monitoring wildlife health and public health once they are in public parks around cities.

Occurrence of antibodies anti -Toxoplasma gondii, Neospora caninum and Leptospira interrogans in a captive deer herd in Southern Brazil.

A large number of Brazilian zoos keep many endangered species of deer, however, very few disease surveillance studies have been conducted among captive cervids. Blood samples from 32 Brazilian deer (Blastocerus dichotomus, Mazama nana and Mazama americana) kept in captivity at Bela Vista Biological Sanctuary (Foz do Iguaçu, Brazil) were investigated for 10 ruminant pathogens, with the aims of monitoring deer health status and evaluating any potential zoonotic risk. Deer serum samples were tested for Brucella abortus, Leptospira (23 serovars), Toxoplasma gondii, Neospora caninum, bovine viral diarrhea virus, infectious bovine rhinotracheitis virus, foot-and-mouth disease virus, western equine encephalitis virus, eastern equine encephalitis virus and Venezuelan equine encephalitis virus. Antibodies against T. gondii (15.6%), N. caninum (6.2%) and L. interrogans serogroup Serjoe (3.1%) were detected. The serological results for all other infectious agents were negative. The deer were considered to be clinically healthy and asymptomatic regarding any disease. Compared with studies on free-ranging deer, the prevalences of the same agents tested among the captive deer kept at the Sanctuary were lower, thus indicating good sanitary conditions and high-quality management practices at the zoo.

Occurrence of antibodies anti -Toxoplasma gondii, Neospora caninum and Leptospira interrogans in a captive deer herd in Southern Brazil / Ocorrência de anticorpos anti -Toxoplasma gondii, Neospora caninum e Leptospira interrogans em um rebanho de cervídeos de cativeiro do sul do Brasil

Abstract A large number of Brazilian zoos keep many endangered species of deer, however, very few disease surveillance studies have been conducted among captive cervids. Blood samples from 32 Brazilian deer (Blastocerus dichotomus, Mazama nana and Mazama americana) kept in captivity at Bela Vista Biological Sanctuary (Foz do Iguaçu, Brazil) were investigated for 10 ruminant pathogens, with the aims of monitoring deer health status and evaluating any potential zoonotic risk. Deer serum samples were tested for Brucella abortus, Leptospira (23 serovars), Toxoplasma gondii, Neospora caninum, bovine viral diarrhea virus, infectious bovine rhinotracheitis virus, foot-and-mouth disease virus, western equine encephalitis virus, eastern equine encephalitis virus and Venezuelan equine encephalitis virus. Antibodies against T. gondii (15.6%), N. caninum (6.2%) and L. interrogans serogroup Serjoe (3.1%) were detected. The serological results for all other infectious agents were negative. The deer were considered to be clinically healthy and asymptomatic regarding any disease. Compared with studies on free-ranging deer, the prevalences of the same agents tested among the captive deer kept at the Sanctuary were lower, thus indicating good sanitary conditions and high-quality management practices at the zoo.

Resumo Um grande número de zoológicos brasileiros abriga espécies de cervídeos ameaçados de extinção, entretanto, estudos de vigilância de doenças em cervídeos de cativeiro são escassos. Amostras de sangue de 32 cervídeos brasileiros (Blastocerus dichotomus, Mazama nana e Mazama americana), mantidos em cativeiro no Refúgio Biológico Bela Vista (Foz do Iguaçu, Brasil), foram investigados para 10 patógenos de ruminantes, visando monitorar o estado de saúde dos cervídeos e avaliar a presença de agentes zoonóticos. As amostras de soro foram testadas para Brucella abortus, Leptospira (23 sorovares), Toxoplasma gondii, Neospora caninum, diarreia viral bovina, rinotraqueíte infecciosa bovina, febre aftosa, encefalomielite equina do oeste, encefalomielite equina do leste e encefalomielite equina venezuelana. Foram detectados anticorpos para T. gondii (15,6%), N. caninum (6,2%) e para L. interrogans sorogrupo Serjoe (3,1%). As sorologias apresentaram resultado negativo para as demais doenças. Os cervídeos foram considerados clinicamente sadios e assintomáticos para doenças. Comparados aos estudos de populações de vida livre, as soroprevalências para os mesmos agentes testados foram menores para os cervídeos mantidos no Refúgio, indicando as boas condições sanitárias e a qualidade das práticas de manejo no zoológico.

Contenção farmacológica da jaguatirica, Leopardus pardalis, para coleta de sêmen, pela associação de tiletamina-zolazepam e xilazina / Chemical restraint of ocelot, Leopardus Pardalis, for semen collection with the association of tiletamine-zolazepam and xylazine

Foram avaliados os efeitos anestésicos da associação de cloridrato de tiletamina (7 mg/kg), cloridrato de zolazepam (7 mg/kg) e cloridrato de xilazina (1 mg/kg), doses calculadas para peso estimado, aplicados por via intramuscular profunda para a contenção farmacológica de machos de jaguatirica, (Leopardus pardalis. Linnaeus, 1758) (Felidae), submetidos à coleta de sêmen por eletroejaculação. Quatro animais foram anestesiados em quatro a cinco ocasiões diferentes, com intervalo mínimo de 30 dias, totalizando 17 procedimentos. Em algumas ocasiões, quando a contenção farmacológica inicial não propiciava segurança para a manipulação do animal, foi necessária a aplicação complementar de anestésicos, sendo então administrada 1/3 da dose inicial, fato necessário em apenas três (17,64%) do total de procedimentos. Após a administração das drogas, foram monitorados durante 80 minutos parâmetros como frequência cardíaca, frequência respiratória, temperatura retal, miorrelaxamento e nocicepção. O período de latência foi de 3,56±0,55 minutos após a injeção e o tempo para recuperação foi de 169,60±12,37 minutos. Não foi evidenciada diferença estatística (p>0,05) entre os animais e os momentos avaliados para reações dolorosas, miorrelaxamento, bem como para os parâmetros: frequências respiratória e cardíaca e temperatura retal. A associação de tiletamina-zolazepam e xilazina, nas doses de 7mg/kg e 1mg/kg respectivamente, proporcionou uma boa qualidade anestésica, curto período de latência, bom período anestésico hábil e com segurança para a realização do procedimento de eletroejaculação nas jaguatiricas, com boa qualidade do sêmen obtido.

The anesthetic effects of tiletamine hydrochloride (7 mg/kg), zolazepam hydrochloride (7 mg/kg) and xylazine (1 mg/kg) association, doses calculated by the estimated weight, were evaluated after deep intramuscular injection for pharmacological restraint of male ocelot (Leopardus pardalis. Linnaeus, 1758) (Felidae), subjected to semen collection by electroejaculation. Four animals were anesthetized on four to five different occasions with an interval of 30 days, totaling 17 procedures. In some instances, when the initial pharmacological restraint was not propitiated safety for handling the animal, it was necessary to apply supplementary anesthetics, and then administered 1/3 of the initial dose, fact necessary only in three procedures (17.64%). After the injection, parameters like heart rate, respiratory rate, rectal temperature, muscle relaxation and nociception was monitored for 80 minutes. The latency period was 3.56±0.55 minutes after injection and the time for recovery was 169.60±12.37 minutes. There was no significant statistical difference (p>0.05) between the animals and moments evaluated for painful reactions, muscle relaxant, as well as for the parameters: respiratory and heart rates and rectal temperature. The combination of tiletamine-zolazepam and xylazine at doses of 1mg/kg and 7mg/kg respectively, provided a good quality of anesthesia, short latency period, good viable anesthetic period and safety to proceed the electroejaculation in ocelots, with good quality of obtained semen.

Mycoplasma ovis in captive cervids: prevalence, molecular characterization and phylogeny.

Hemotrophic mycoplasmas (hemoplasmas) are bacteria that attach to red blood cells of mammals, leading to acute and/or subclinical disease in infected animals. It has been suggested that Mycoplasma ovis, a hemoplasma that infects sheep and goats worldwide, may also infect deer. The aim of this study was to evaluate whether South American deer are infected with M. ovis. EDTA-anticoagulated blood samples from a herd of 32 captive South American deer were collected. DNA extraction of blood samples was performed followed by PCR amplification of the 16S and 23S rRNA genes, and sequencing of products. Using M. ovis PCR, 27/31 (87%) were positive, including 21/22 Mazama nana; 2/3 Mazama americana and 4/6 Blastocerus dichotomus. Sequencing of the nearly entire 16S rRNA gene of 26/27 positive samples showed 98.2-98.8% identity to M. ovis of sheep (GenBank, AF338268) and 98.6-99.4% identity to M. ovis-like of a fawn (FJ824847); the 23S rRNA gene from one of these isolates and the fawn's had 97.6% identity. The remaining isolate had just 94.9% identity to the 16S rRNA gene of M. ovis and only 89.4% identity to the 23S rRNA gene of the fawn's M. ovis. This is the first report of M. ovis in captive South American deer, revealing a high prevalence of hemoplasma infection in these animals.

Use of a Mycoplasma suis-PCR protocol for screening a population of captive peccaries (Tayassu tajacu and Tayassu pecari).

Mycoplasma suis is a hemotropic bacteria of red blood cells and the causative agent of swine eperythrozoonosis. Diagnosis of infection may be reached by direct examination of blood smears; however, the use of polymerase chain reaction (PCR) of the 16S RNA gene of M. suis improves the sensitivity and specificity of detection. The aim of this study was to screen peccaries (Tayassu tajacu and T. pecari) for M. suis infection using a specific conventional PCR. A total of 28 blood samples from captive collared and white-lipped peccaries were collected, DNA extracted and a specific M. suis PCR assay performed. All samples were negatives by both blood smear examination and PCR testing. To verify the presence of amplifiable DNA, PCR for beta-actin gene was performed in all samples. This study was part of an active surveillance program, which is crucial for monitoring animal health status, particularly in wildlife species.