Equine Rabies in the Southern Region of Piauí State

Background: Rabies is an infectious disease that is important in the "One Health" worldwide with high lethality rate. The etiological agent is a neurotropic virus, genus Lyssavirus, transmitted mainly through the saliva of infected animals. For equines, the bite of hematophagous bats is the main source of infection. Piauí is an important state for equestrian sports and the increase in the number of horses with neurological clinical signs without diagnosis has increased in recent years. In this context, the aim of this study is to report to the scientific community a confirmed case of equine rabies in the Santa Luz county, Southernmost state of Piauí, Brazil.Case: A 3-year-old female non-defined breed horse, was admitted to the Hospital Veterinário da Universidade Federal do Piauí (UFPI/CPCE). The equine had difficulty walking 2 days ago, in the panoramic inspection was restless and disoriented in the paddock. Rectal temperature of 38.2oC, heart rate of 60 bpm, respiratory rate of 40 mpm, congested mucosa and dyspnea were verified. With the progression of the neurological signals, it positioned itself in a lateral decubitus with pedaling movements, hyperesthesia, dysphagia and paralysis of the hindlimbs. The clinical suspicion was rabies and the Agência de Defesa Agropecuária do Piauí (ADAPI) was communicated to euthanize the animal and collect samples for diagnosis in accordance with official standards of the Ministério da Agricultura, Pecuária e Abastecimento (MAPA). At necropsy, there was slight brain hyperemia, with no other significant organ changes. Fragments of the cerebellum, cortex, hippocampus and spinal cord were collected and sent at a temperature of 4oC to perform the Direct Immunofluorescence (DIF) assay. Samples for histopathology were not collected because they do not include assay for confirmatory diagnosis of rabies. The DIF technique with antigen-labeled antibodies were performed in the imprint lamina of these fragments. The fragments were treated according to specific protocol. The results were negative for DIF in the collected equine fragments. For complementary exam, the samples were homogenized, clarified and inoculated intracranial in BALB/C mice, being observed for up to 30 days. The samples were positive after the bioassay.Discussion: Piauí is a state with great equestrian activity that expose the animals to the risks of transmission of infectious diseases. Among these diseases, rabies is important for affecting horses, but also humans (veterinarians and owners). In the present report, the equine showed clinical signs of furious rabies for a short period, rapidly evolving to paralytic form. This clinical aspect must be carefully evaluated by the veterinarian, in order to avoid false clinical suspicions such as tetanus or other non-infectious diseases. The official diagnosis of the rabies is DIF technique, with high sensitive (80-100%). According to the results of the DIF technique, it was possible to confirm the clinical suspicion of rabies in mice previously inoculated with emulsion of fragments of the equine central nervous system (CNS). This fact demonstrates that in negative results for CNS samples from the horse, the bioassay increases the sensitivity of the test and avoids false negative diagnoses. Thus, it was possible to prove that rabies is affecting the equines in the southern region of Piauí state and alerts the breeders and the community to intensify surveillance and control of the hematophagous bats. For the authors, this is the first scientific report of rabies in the region studied

Isolation, Expansion, Differentiation and Growth Kinetics Essay in Mesenchymal Stem Cells Culture from the Bone Marrow of Collared Peccaries (Tayassu tajacu)

Background: There are few studies on stem cell isolation in wild animals that provide isolation and culture protocols of these cells in vitro. Among the wild species studied, we present the collared peccary (Tayassu tajacu) as a model with potential to obtain and use MSC in preclinical studies. These animals are phylogenetically close to the domestic pig, popularly known as peccaries and found naturally in South America, Central America and the South of the United States. The aim of the present study was to establish a protocol for the isolation, in vitro cell expansion, differentiation and assessment of the stromal MSC growth curve before and after thawing.Materials, Methods & Results: Mesenchymal stem cells (MSC) from collared peccary bone marrow (Tayassu tajacu) were isolated and expanded by centrifuge in Ficoll® solution and cultured in DMEM® High Glucose medium. The culture was assessed by assays of colony forming units CFU-F and growth curve by saturation (GCS). Cultures in the third passage, with 70% confluence, were replicated at 105 cells/mL concentration in the culture media to induce osteogenic cell differentiation and adipogenic cell differentiation, respectively. The MSC were frozen in nitrogen for 40 days, thawed and re-assessed for cell viability and GCS.Discussion: The bone marrow collected presented high mononuclear cellularity, with a mean variability of 94.5% and 60.83 ± 4.27 UFC were identified in the samples and cells with fibroblast-like-cell morphology were observed. When they were expanded, the mean cell viability was 95%, the mean cell concentration obtained was 233.31 ± 20.04 cells per 25cm2 bottle and the culture reached the growth plateau in GCS between the 13th and 16th day. The osteoblastic cell differentiation assay showed after 18 days, morphology similar to osteoblasts, with irregular cytoplasm limits, cell prolongation formation and flattened appearance. After staining with Alizarin Red, the nucleus presented a wine red coloring and the cytoplasm, more basophilic and well-defined, with calcium deposits inside the cells. The cultures submitted to adipogenic differentiation were large, hexagonal, irregular and presented birrefringent cytoplasm granules after the third week of culture. When stained with Oil Red it was observed that the cytoplasm granules were scattered small fat vacuoles and stained maroon. The viability after thawing was 78% and the mean cell concentration obtained in GCS was 199.71 ± 14.72 cells per 25 cm2 bottle. The curves reached the saturation plateau early, on the eighth day of observation. From then onwards the cultures entered became exhausted and the cell concentration of the samples decreased progressively until minimum values. These results showed the presence of a well-defined MSC population in the collared peccary bone marrow with a high rate of replication in vitro and potential for differentiation confirmed by the adipogenic and osteogenic lines. The cryopreservation technique adopted presented satisfactory results, but indicated a significant cell stress after thawing that justifies investigation of the apoptosis rates induced post thawing in the species. Furthermore, the bone marrow collection did not harm the animals and the facility of stromal MSC isolation and culture qualifies the collared peccary as a viable alternative model to obtain MSC and for studies in the area of cell therapy

In vitro maturation of agoutis (Dasyprocta prymnolopha) oocytes followed by in vitro fertilization and parthenogenetic activation

O objetivo foi avaliar protocolos de maturação in vitro (MIV) para oócitos de cutias, seguida de fertilização in vitro (FIV) e ativação partenogenética (AP). Os oócitos imaturos (CCOs) foram obtidos por fatiamento do ovário, após OSH, e submetidos a três grupos: MAT - 16 (16 horas de maturação), MAT - 20 (20 horas de maturação) e MAT - 24 (24 horas de maturação), em incubadora de cultivo a 38,8°C, com atmosfera de 5% de CO2 e 95% de umidade relativa. A maturação foi analisada pela presença do primeiro corpúsculo polar. Em seguida, os CCOs maduros foram submetidos à FIV, com período de coincubação dos CCOs e dos espermatozoides de 15h, a 38,8ºC e 5% de CO2, e AP com ionomicina. Os grupos de MIV foram analisados utilizando-se o teste qui-quadrado e, nos experimentos de FIV e AP, foram analisadas a taxa de clivagem e a proporção de desenvolvimento embrionário. A análise estatística foi realizada utilizando-se o programa SAS. Houve diferença significativa entre os grupos de maturação, tendo os grupos MAT - 20 e MAT - 24 apresentado maior porcentagem de oócitos maturados in vitro. As taxas de clivagem e de desenvolvimento embrionário foram de 8,6% e 2,9%, respectivamente, na FIV, e de 63,6% e 15,1%, na AP. Entretanto, nos dois casos, o embrião não passou do estágio de mórula.The objective was to evaluate IVM protocols for agouti oocytes, followed by in vitro fertilization (IVF) and parthenogenetic activation (PA). The immature oocytes (CCOs) were obtained by slicing the ovary after OSH and submitted to three groups: MAT - 16 (16 hours maturation), MAT - 20 (20 hours maturation) and MAT – (24 hours maturation), in a culture incubator at 38.8°C, with an atmosphere of 5% CO2 and 95% relative humidity. The maturation was analyzed by the presence of the first polar corpuscle. Then, mature CCOs were submitted to IVF, with co-incubation period of CCOs and spermatozoa from 15h to 38.8°C and 5% of CO2, and PA with inomycin. The IVM groups were analyzed using the chi-square test and in the FIV and PA experiment the rate of cleavage and the rate of embryonic development were analyzed. Statistical analysis was performed using the SAS program. There was a significant difference between the maturation groups, and the MAT - 20 and MAT - 24 groups showed a higher percentage of matured oocytes in vitro. The rates of cleavage and embryonic development were 8.6% and 2.9%, respectively in FIV and 63.6% and 15.1% in PA. However, in both cases the embryo did not pass beyond the morula stage

AMPUTAÇÃO DO MEMBRO PELVICO ESQUERDO DE TAMANDUÁ-MIRIM (Tamanduá tetradactyla) (relato de caso) AMPUTATION OF THE LEFT PELVIC MEMBER OF LITTLE ANTEATER (Tamandua tetradactyla) (case report)

Os traumas são comuns em animais silvestres e correspondem a 15,5% das desordens clínicas registradas em tamanduás. No entanto, há poucos relatos de procedimentos cirúrgicos em animais silvestres. Este relato visa contribuir com informações sobre o tratamento cirúrgico utilizado para a amputação do membro pélvico de um tamanduá-mirim (Tamandua tetradactila), realizado no Hospital Veterinário Universitário da Universidade Federal do Piauí, em decorrência de um trauma extenso com perda parcial da extremidade distal do membro pélvico. O procedimento cirúrgico favoreceu a sobrevivência e o completo restabelecimento do animal. <br /><br />PALAVRAS-CHAVES: Animal silvestre, cirurgia, Tamandua tetradactyla. Trauma is common in wild animals. Correspond to 15.5% of the clinical disorders registered in anteaters. However, reports of surgical procedures in wild animals not are commons. This report describes amputation of the pelvic member of a little anteater (Tamandua tetradactila), performed at the Universitary Veterinary Hospital of the Federal University of Piaui, due to extensive trauma. This case is very important because the animal survived to surgical stress and presented complete recuperation.<br /><br />KEY WORDS: Surgery, Tamandua tetradctyla, wild animal.<br /&gt

Evaluation of hematological profile, biochemical and peripheral blood smear with a view to the health profile in primates of the Cebus genre maintained in captivity

Os primatas não humanos atuam como modelo para estudos sobre a dinâmica das vias de transmissão e história natural de doenças compartilhadas entre homens e animais. O ambiente de cativeiro é propício à disseminação de doenças de caráter zoonótico. Muitos destes animais não apresentam sintomatologia clínica, mesmo estando infectados, o que os caracterizam como importante fonte de infecção para os animais domésticos e o homem. O objetivo deste trabalho foi avaliar a sanidade de primatas não humanos mantidos em cativeiro através de análise hematológica e bioquímica, bem como esfregaço de sangue periférico, com intuito de investigar a presença de patógenos de caráter zoonótico, servindo de modelo para estudos futuros sobre a dinâmica das vias de transmissão de doenças compartilhadas entre homens e animais. Foram coletadas amostras de sangue de 15 macacos prego (Cebus sp.), adultos, clinicamente saudáveis e pertencentes ao Parque Zoobotânico de Teresina. Foram coradas lâminas de esfregaço sanguíneo e obtidos os perfis hematológicos e bioquímicos de cada animal. A análise dos dados baseou-se em estatística básica. Não foi observado nenhum hemoparasita presente em sangue periférico. Todos os animais estavam anêmicos, 46,7% trombopênicos e 87% apresentavam algum tipo de processo patológico de evolução crônica, devido à elevada taxa de monócitos encontrados. Todos os animais apresentaram elevadas taxas de fosfatase alcalina e das transaminases AST e ALT, indicando injúria do parênquima hepático. Novos estudos deverão ser realizados para melhor elucidação dos resultados, visto que dados bioquímicos fisiológicos de primatas do gênero Cebus são escassos na literatura.Non-human primates serve as a model for studies on the dynamics of transmission routes and natural history of diseases shared between humans and animals. The captive environment is conducive the dissemination of zoonotic diseases. Many of these animals do not present clinical symptomatology, even when infected, which characterize them as an important source of infection for domestic animals and man. The objective of this study was to evaluate the sanity of non-human primates kept in captivity through hematological and biochemical analysis as well as peripheral blood smear, aiming to investigate the presence of zoonotic pathogens, serving as a model for future studies on the dynamics of routes of transmission of diseases shared between humans and animals. Were collected samples of blood from 15 nail monkeys (Cebus sp.), adults, clinically healthy and belonging to Park Zoobotanic of Teresina. Were stained smear sanguine blades and obtained the haematological and biochemical profiles of each animal. The data analysis was based on basic statistics. Did not observed any haemoparasite present in peripheral blood. All animals were anemic, 46,7% thrombopenics and 87% of the animals showed some type of pathological process of chronic evolution, due to the high rate of monocytes found. All animals showed high rates of alkaline phosphatase, and transaminases AST and ALT, indicating injury of the hepatic parenchyma. New studies should be conducted to better elucidate of results, seeing that biochemical physiological data primate of the genus Cebus are scarce in literature.

In vitro maturation of agoutis (Dasyprocta prymnolopha) oocytes followed by in vitro fertilization and parthenogenetic activation

O objetivo foi avaliar protocolos de maturação in vitro (MIV) para oócitos de cutias, seguida de fertilização in vitro (FIV) e ativação partenogenética (AP). Os oócitos imaturos (CCOs) foram obtidos por fatiamento do ovário, após OSH, e submetidos a três grupos: MAT - 16 (16 horas de maturação), MAT - 20 (20 horas de maturação) e MAT - 24 (24 horas de maturação), em incubadora de cultivo a 38,8°C, com atmosfera de 5% de CO2 e 95% de umidade relativa. A maturação foi analisada pela presença do primeiro corpúsculo polar. Em seguida, os CCOs maduros foram submetidos à FIV, com período de coincubação dos CCOs e dos espermatozoides de 15h, a 38,8ºC e 5% de CO2, e AP com ionomicina. Os grupos de MIV foram analisados utilizando-se o teste qui-quadrado e, nos experimentos de FIV e AP, foram analisadas a taxa de clivagem e a proporção de desenvolvimento embrionário. A análise estatística foi realizada utilizando-se o programa SAS. Houve diferença significativa entre os grupos de maturação, tendo os grupos MAT - 20 e MAT - 24 apresentado maior porcentagem de oócitos maturados in vitro. As taxas de clivagem e de desenvolvimento embrionário foram de 8,6% e 2,9%, respectivamente, na FIV, e de 63,6% e 15,1%, na AP. Entretanto, nos dois casos, o embrião não passou do estágio de mórula.The objective was to evaluate IVM protocols for agouti oocytes, followed by in vitro fertilization (IVF) and parthenogenetic activation (PA). The immature oocytes (CCOs) were obtained by slicing the ovary after OSH and submitted to three groups: MAT - 16 (16 hours maturation), MAT - 20 (20 hours maturation) and MAT – (24 hours maturation), in a culture incubator at 38.8°C, with an atmosphere of 5% CO2 and 95% relative humidity. The maturation was analyzed by the presence of the first polar corpuscle. Then, mature CCOs were submitted to IVF, with co-incubation period of CCOs and spermatozoa from 15h to 38.8°C and 5% of CO2, and PA with inomycin. The IVM groups were analyzed using the chi-square test and in the FIV and PA experiment the rate of cleavage and the rate of embryonic development were analyzed. Statistical analysis was performed using the SAS program. There was a significant difference between the maturation groups, and the MAT - 20 and MAT - 24 groups showed a higher percentage of matured oocytes in vitro. The rates of cleavage and embryonic development were 8.6% and 2.9%, respectively in FIV and 63.6% and 15.1% in PA. However, in both cases the embryo did not pass beyond the morula stage

Isolation, Expansion, Differentiation and Growth Kinetics Essay in Mesenchymal Stem Cells Culture from the Bone Marrow of Collared Peccaries (Tayassu tajacu)

Background: There are few studies on stem cell isolation in wild animals that provide isolation and culture protocols of these cells in vitro. Among the wild species studied, we present the collared peccary (Tayassu tajacu) as a model with potential to obtain and use MSC in preclinical studies. These animals are phylogenetically close to the domestic pig, popularly known as peccaries and found naturally in South America, Central America and the South of the United States. The aim of the present study was to establish a protocol for the isolation, in vitro cell expansion, differentiation and assessment of the stromal MSC growth curve before and after thawing.Materials, Methods & Results: Mesenchymal stem cells (MSC) from collared peccary bone marrow (Tayassu tajacu) were isolated and expanded by centrifuge in Ficoll® solution and cultured in DMEM® High Glucose medium. The culture was assessed by assays of colony forming units CFU-F and growth curve by saturation (GCS). Cultures in the third passage, with 70% confluence, were replicated at 105 cells/mL concentration in the culture media to induce osteogenic cell differentiation and adipogenic cell differentiation, respectively. The MSC were frozen in nitrogen for 40 days, thawed and re-assessed for cell viability and GCS.Discussion: The bone marrow collected presented high mononuclear cellularity, with a mean variability of 94.5% and 60.83 ± 4.27 UFC were identified in the samples and cells with fibroblast-like-cell morphology were observed. When they were expanded, the mean cell viability was 95%, the mean cell concentration obtained was 233.31 ± 20.04 cells per 25cm2 bottle and the culture reached the growth plateau in GCS between the 13th and 16th day. The osteoblastic cell differentiation assay showed after 18 days, morphology similar to osteoblasts, with irregular cytoplasm limits, cell prolongation formation and flattened appearance. After staining with Alizarin Red, the nucleus presented a wine red coloring and the cytoplasm, more basophilic and well-defined, with calcium deposits inside the cells. The cultures submitted to adipogenic differentiation were large, hexagonal, irregular and presented birrefringent cytoplasm granules after the third week of culture. When stained with Oil Red it was observed that the cytoplasm granules were scattered small fat vacuoles and stained maroon. The viability after thawing was 78% and the mean cell concentration obtained in GCS was 199.71 ± 14.72 cells per 25 cm2 bottle. The curves reached the saturation plateau early, on the eighth day of observation. From then onwards the cultures entered became exhausted and the cell concentration of the samples decreased progressively until minimum values. These results showed the presence of a well-defined MSC population in the collared peccary bone marrow with a high rate of replication in vitro and potential for differentiation confirmed by the adipogenic and osteogenic lines. The cryopreservation technique adopted presented satisfactory results, but indicated a significant cell stress after thawing that justifies investigation of the apoptosis rates induced post thawing in the species. Furthermore, the bone marrow collection did not harm the animals and the facility of stromal MSC isolation and culture qualifies the collared peccary as a viable alternative model to obtain MSC and for studies in the area of cell therapy