Detección de anticuerpos anti-Mycobacterium avium subsp. paratuberculosis (MAP) en venados silvestres en Colombia / Detection of anti-Mycobacterium aviumsubsp. paratuberculosis antibodies in wild deer in Colombia

RESUMEN Objetivo. Determinar la presencia de anticuerpos anti-Mycobacterium avium subsp. paratuberculosis (MAP) en venados de cola blanca (Odocoileus virginianus) y venados de páramo (Mazama rufína), capturados en las regiones de la Orinoquía y el Caribe en Colombia. Materiales y métodos. Un total de 44 muestras de suero sanguíneo de venados de las especies M. rufína y O. virginianus fueron colectadas en condiciones de campo entre 2014 y 2016. Se utilizó un kit comercial de ELISA para la detección de anticuerpos anti-MAP. Un animal se consideró positivo a ELISA cuando la relación muestra-positivo (S/P%) fue > 0.4, según lo recomendado por el fabricante. Resultados. El 50% (22/44) de los animales muestreados resultaron positivos, lo que corresponde a 10 hembras y 12 machos. Un 81.8% (18/22) y un 77.3% (17/22) de estos animales seropositivos fueron capturados en la región de la Orinoquía y fueron reportados como adultos, respectivamente. Conclusiones. No se sabe cómo o cuándo se introdujo MAP en la población de ciervos colombianos en las regiones de estudio. La hipótesis más plausible para explicar la presencia de anticuerpos anti-MAP en estas poblaciones silvestres es la transmisión por contacto con el ganado bovino infectado, ya que en ambas regiones estas especies comparten pasturas. Este es el primer estudio en explorar la infección por MAP en animales silvestres en Colombia. Estos hallazgos respaldan la necesidad de realizar más estudios utilizando técnicas de diagnóstico directo, y aproximaciones investigativas que permitan la definición de vínculos en la dinámica de la infección entre mamíferos silvestres y domésticos en Colombia.

ABSTRACT Objective. To determine the presence of anti-Mycobacterium avium subsp. paratuberculosis (MAP) antibodies in white-tailed deer (Odocoileus virginianus) and red brocket deer (Mazama rufína), captured in the Orinoquía and Caribbean regions in Colombia. Materials and methods. A total of 44 blood serum samples from deer of species M. rufina and O. virginianus were collected under field conditions between 2014 and 2016. An ELISA commercial kit was used to detect anti-MAP antibodies. An animal was considered ELISA-positive at a sample-to-positive ratio (S/P%) of > 0.4, as recommended by the manufacturer. Results. The 50% (22/44) of the animals were positive, corresponding to 10 females and 12 males. An 81.8% (18/22) and 77.3% (17/22) of these seropositive animals were captured in the Orinoquía region and were reported as adults, respectively. Conclusions. It is not known how or when MAP was introduced in the Colombian deer population in the study regions. The most plausible hypothesis to explain the presence of antibodies against MAP in these wild populations is transmission by contact with infected bovine cattle since, in both regions, these species share pastures. This is the first study to explore MAP infection in wild animals in Colombia. These findings support the need for further studies using different direct diagnostic techniques and research approaches that allow the definition of links in the infection dynamics between wild and domestic mammals in Colombia.

First report of complete albinism in Mazama americana (Erxleben, 1777) in the Biological Reserve of Tapirapé, Oriental Amazon, Brazil

Albinism is a genetic condition that results in total hypopigmentation of the eyes, fur, skin, hair, scales, and feathers of an organism. Albinism might result in a selective disadvantage for affected animals. Cases of albinism have been previously recorded in Neotropical vertebrates, such as reptiles, mammals, birds, and fish. However, observing albinism in a wild population is still considered to be a rare event. This paper reports a unique case of complete albinism in a red-brocket deer (Mazama americana) living in the Brazilian Amazon rainforest. The individual was observed within the Biological Reserve of Pará State, one of the most deforested regions of the Brazilian Amazon. The survival of the albino red-brocket deer in the wild can be related to mechanisms of apostatic selection, which theorize the survival of individual prey animals whose mutations make them less likely to be attacked by predators. In other words, the more different a prey animal is from others, the less likely it will be targeted by predators. The high abundance prey animals within the Biological Reserve of Tapirapé seems to support this prediction. This report exemplifies the importance of monitoring the biodiversity and promoting the conservation of favorable habitats to support species multiplicity in highly fragmented regions, as in the Brazilian Amazon.

CWD in Norway

The Norwegian Food Safety Authority (NFSA) and Norwegian Environmental Authority (NEA) asked the Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) for an opinion on factors associated with the introduction of Chronic Wasting Disease (CWD) to Norway. VKM appointed a working group consisting of two members of the Panel on Biological Hazards, one member of Panel on Animal Health and Welfare, and two external experts to prepare the answer to the questions. The Panel on Biological Hazards has reviewed and revised the draft prepared by the working group and approved the opinion. CWD was diagnosed in March 2016 in a wild reindeer (Rangifer tarandus) from the Nordfjella mountain area in Norway and in May and June in two mooses (Alces alces) in Selbu in South Trøndelag County, approximately 300 km north from the first case. There is currently no information to determine the origin(s) of CWD agents in Norway. However, the sporadic or genetic (somatic mutation) occurrence of prion disease in cervids cannot be excluded, nor can introduction from North America or other countries. Furthermore, there is no evidence that it has not been circulating at low levels in the Norwegian cervid populations for years, but has not previously been identified. In this scientific opinion, information on prion diseases in general, and CWD in particular, is presented in the light of experiences with this disease in North America. Prions are among the most resilient pathogens known and dissemination of prions into ecosystems is likely to result in long-term problems. Prions bind strongly to soil and remain infectious. In CWD, prions are present in most peripheral organs and also shed into the environment via saliva, faeces, and urine, as well as with the placenta. CWD transmits easily among cervids, either through direct contact, or indirectly via the environment. Migration of animals is relevant for the spread between areas. Strain diversification might occur in CWD and may influence transmission properties of the agents. Clinical signs of CWD are non-specific and do not alone enable confirmation of the diagnosis. Analysis of tissue from the brainstem at the level of the obex by approved methods is necessary for diagnosis of CWD. Prion infectivity is assessed by bioassays, often involving transgenic mice. In vitro conversion assays, like protein misfolding cyclic amplification (PMCA), provide sensitive quantification of converting activity, which is a good approximation of infectivity. Genetic variation (polymorphisms) in the gene that encodes PrP (PRNP) can modulate sensitivity towards CWD. The level of such genetic variation in Norwegian wild and semi-domesticated cervids is currently unknown. Cattle and sheep are at very low risk of developing CWD and it is highly unlikely that prion diseases in sheep or cattle are the origin of CWD. Although transmission of CWD to humans has never been known to occur, and animals other than cervids have not been found to be infected, indicating a species barrier, this possibility cannot be excluded. Thus, measures for reduction of human exposure are recommended. Taking into account uncertainties regarding the plasticity of the CWD agents and the lack of transmission data from the Norwegian isolates, this scientific opinion considers the zoonotic risk of CWD to be very low.

Detection of bluetongue virus in Brazilian cervids in São Paulo state / Detecção de vírus da língua azul em cervídeos brasileiros no estado de São Paulo

Viral hemorrhagic diseases in cervids occur worldwide and include epizootic hemorrhagic disease (EHD), bluetongue (BT), and adenoviral hemorrhagic disease (AHD). Since gross lesions in all three hemorrhagic diseases are identical (hemorrhagic enteropathy, pulmonary edema, systemic petechial and suffusion hemorrhages), it is necessary to use accurate techniques for a definitive etiologic diagnosis. Archival material (paraffin blocks) at the Department of Veterinary Pathology of FCAV - Unesp was reviewed for lesions of hemorrhagic disease and 42 captive and free-living Brazilian deer were selected to include in this study. Paraffin-embedded tissues were evaluated using immunohistochemistry and tested negative for adenovirus. Using real time RT-PCR, EHD virus was not detected in paraffin-embedded tissues in any of the cases evaluated. The same technique was used for detection of BT virus and seven positive animals (16,66%) were confirmed after agarose 4% gel electrophoresis and gene sequencing. The main macroscopic changes observed in the positive animals were hemorrhagic intestinal contents, reddish mucous membrane of the gastrointestinal tract, ulcers on tongue and petechiae in various organs. Microscopic changes observed were lymphocytic inflammatory infiltrate in liver, kidney and lungs, hemorrhage, and congestion in various organs. All positive cases were from captive animals, three females (two young and one adult), and four young males. This study demonstrates that the bluetongue virus is involved in hemorrhagic disease outbreaks of deer in Brazil.(AU)

Doenças hemorrágicas virais em cervídeos ocorrem no mundo todo e incluem a doença epizoótica hemorrágica (DEH), língua azul (LA), e doença hemorrágica por adenovírus (DHA). Uma vez que as lesões nas três doenças hemorrágicas são idênticas (enteropatia hemorrágica, edema pulmonar, petéquias sistêmicas e sufusões hemorrágicas), é necessário utilizar técnicas precisas para um diagnóstico etiológico definitivo. Material de arquivo (blocos de parafina) do Departamento de Patologia Veterinária da FCAV - Unesp foi revisado para lesões de doenças hemorrágicas e 42 cervídeos brasileiros de cativeiro e de vida livre foram selecionados e incluídos neste estudo. Tecidos embebidos em parafina foram avaliados usando imunohistoquímica e foram negativos para adenovírus. Usando o RT-PCR em tempo real, o vírus da DEH não foi detectado nos tecidos de nenhum dos casos avaliados. A mesma técnica foi utilizada para detecção do vírus da LA e sete animais positivos (16,66%) foram confirmados após eletroforese em gel de agarose a 4% e sequenciamento genético. As principais alterações macroscópicas observadas nos animais positivos foram conteúdo intestinal hemorrágico, mucosa do trato gastrointestinal avermelhada, úlceras na língua e petéquias em vários órgãos. As alterações microscópicas observadas foram infiltrado inflamatório linfocítico em fígado, rins e pulmões, e hemorragia e congestão em vários órgãos. Todos os casos positivos foram de animais de cativeiro, três fêmeas (dois jovens e um adulto), e quatro jovens do sexo masculino. Este estudo demonstra que o vírus da lingual azul está envolvido nos surtos de doença hemorrágica em veados no Brasil.(AU)

Osteología del miembro torácico de la corzuela parda Mazama gouazoubira (g. Fischer, 1814) (Cetartiodactyla: Cervidae) / Osteology of the thoracic limb of south american gray brocket Mazama gouazoubira (Fischer, 1814) (Cetartiodactyla: Cervidae)

El objetivo del estudio fue describir las características anatómicas y establecer hipótesis morfo-funcionales del esqueleto del miembro torácico de Mazama gouazoubira. Para ello, se utilizaron los miembros torácicos de cuatro adultos jóvenes. Los huesos se prepararon mediante ebullición y fueron blanqueados en una solución de peróxido de hidrógeno para obtener descripciones comparativas, mediciones osteométricas y radiografías digitales. En comparación con los rumiantes domésticos, la escápula resultó ser ancha y plana, con el acromion pequeño, el proceso coracoides muy pequeño y sin tuberosidad de la espina. El húmero se observó alargado con la diáfisis redondeada en una sección transversal y con proyecciones no articulares lisas. La ulna se unió al radio hasta la epífisis distal y juntos formaron un espacio interóseo proximal alargado y estrecho, sin surco vascular. El esqueleto de la mano presentó los metacarpianos III y IV bien desarrollados y fusionados, mientras que los metacarpianos II y V resultaron rudimentarios con disposición telemetacarpiana, filogenéticamente típico de cérvidos del Nuevo Mundo. Se encontraron cuatro dedos con tres falanges en cada uno, de los cuales, dos corresponden a los principales (III y IV) que llegan al suelo y dos son rudimentarios (II y V). Las radiografías nos permitieron visualizar los patrones de tensiones trabeculares normales y la osteometría permitió establecer relaciones con fines comparativos. Se reconocieron las adaptaciones esqueléticas del miembro torácico para favorecer la locomoción cursorial saltatoria.

This study aimed to describe anatomical features of the forelimb skeleton of Mazama gouazoubira in order to establish morphofunctional activity. Forelimbs of four young adult specimens were used to this end. The bones were prepared by boiling and cleared in a solution of hydrogen peroxide for comparative descriptions, osteometrics measurements and digital radiographs. Compared to domestic ruminants, the scapula proved to be wide and flat, with a small acromion, reduced coracoid process and lacking of spine tuberosity. The humerus had an elongated shaft with a rounded cross-section and discrete non-articular projections. The ulna accompanied the radio to the distal epiphysis and formed a long and narrow, proximally placed interosseous space without a vascular groove. The skeleton of the hand presented the rudimentary metacarpals II and V and the well-developed III and IV fused metacarpals, which is a typical telemetacarpal arrangement of close phylogenetically deer species from the New World. There were four fingers, each one with three phalanges; two main (III and IV) touching the ground and two rudimentary (II and V). Radiographs allowed visualizing patterns of normal trabecular tensions and osteometrics enabled to establish ratios for comparative purposes. Adaptations of skeletal forelimb to favor cursorial saltatory locomotion were typically identified.

Avaliação de dois métodos para condicionamento e coleta de sêmen em quatro espécies do gênero Mazama / Assessment of two methods for conditioning and semen collection in four species of Mazama genus

O desenvolvimento de técnicas não invasivas para a obtenção de sêmen de cervídeos facilita a criação de bancos genômicos, que são importantes instrumentos para a conservação ex situ e in situ. Este trabalho teve como objetivo criar uma metodologia não-invasiva de coleta de sêmen e comparar duas técnicas de coleta em quatro espécies do gênero Mazama: M. americana, M. gouazoubira, M. nana e M. nemorivaga. Para tanto, foram utilizados seis machos (M) e duas fêmeas (F) da espécie M. americana, 3M e 2F de M. gouazoubira, 1M e 1F de M. nana e 2M e 1F de M. nemorivaga. Para cada técnica testada, foi realizado um período de habituação dos animais ao manejo. Em seguida, duas técnicas de condicionamento e coleta foram avaliadas. Na primeira delas foi utilizada uma fêmea em estro com desvio lateral do pênis para vagina artificial (FEDL), obtendo-se a coleta de 50% dos indivíduos (100% dos machos de M. gouazoubira e 50% dos machos de M. americana), não obtendo ejaculados das demais espécies. Na segunda técnica, utilizando um manequim taxidermizado com urina de fêmea em estro (MUFE) não foi possível a coleta de nenhum ejaculado. Em todas as fases foi observado o comportamento do macho quanto ao tempo de interesse e aproximação, reflexo de "Flehmen", ato de cheirar ou lamber, exposição do pênis, ereção, número de falsas montas, tentativas de cópula e ocorrência de agressividade entre os animais.

The development of noninvasive techniques for obtaining semen from deer facilitates the creation of genome banks, which are important tools for ex situ and in situ conservation. This study aimed to establish a noninvasive method of semen collection and compare two techniques of collection in four species of the genus Mazama: M. americana, M. gouazoubira, M. nana and M. nemorivaga. To achieve this, 6 males (M) and 2 females (F) of the species M. Americana, 3M and 2F of M. gouazoubira, 1M and 1F of M. nana and 2M and 1F of M. nemorivaga were used. For each technique tested, a period of habituation to animal handling was conducted; then, the two conditioning techniques and collection were evaluated. In the first, a female in estrus was used with lateral deviation of the penis to an artificial vagina (FEDL), yielding collection from 50% of the males (100% from M. gouazoubira and 50% from M. americana), with no ejaculate from the remaining species. In the second technique, using a taxidermized dummy with urine from females in estrus (MUFE), no semen collection was possible. During all stages, male behavior was observed regarding the time of interest and approximation, the "Flehmen" response, the act of sniffing or licking, exposure of the penis, erection, number of false mounts, attempts at copulation and the occurrence of aggression between the deer.

Effects of laparoscopy on the cardiorespiratory system of brown brocket deer (Mazama gouazoubira) anesthetized with ketamine/ xylazine/ midazolam combination and isoflurane / Efeitos da laparoscopia sobre o sistema cardiorrespiratório de veados-catingueiro (Mazama gouazoubira) anestesiados com a associação cetamina/xilazina/midazolam e isofluorano

Laparoscopy is not widely used as a tool to perform assisted reproduction techniques in South American cervids; thus, scarce information in literature is available regarding its effects and appropriate anesthetic protocols to perform it. This study evaluated the effect of laparoscopy on heart rate (HR), respiration rate (RR), saturation of oxyhemoglobin (SpO2) and rectal temperature (RT) of six female brown brocket deer (Mazama gouazoubira) anesthetized with ketamine (5mg/kg), xylazine (0.3mg/kg), midazolam (0.5mg/kg) combination i.v. and isoflurane. Twelve laparoscopies were performed and each animal was used twice with a 40-day interval. After anesthetized, the animals were placed in dorsal recumbency to perform laparoscopy procedure using abdominal CO2 insufflations (14.2 ± 2.39mmHg; M ± SE). The main events of the laparoscopy procedure were divided into three periods: animal without (P1) and with abdominal insufflation (P2) and abdominal insufflation with the hips raised at 45º (P3). As a control, the animals were anesthetized again 40 days after the last laparoscopy, and were maintained in a dorsal recumbency for the same average duration of the previous anesthesia and no laparoscopy procedure was conducted. The period of anesthesia for the controls was also divided into P1, P2, and P3 considering the average duration of these periods in previous laparoscopies performed. Data were analyzed through the (ANOVA) variance analysis followed by Tukey test and values at P<0.05 were considered significant. No significant differences were observed in the parameters evaluated at P1, P2 and P3 between the animals submitted to laparoscopy and control. However, the RR mean between P1 (38.8 ± 4.42) and P3 (32.7 ± 4.81); and the RT mean between the P1 (38.2ºC ± 0.17), P2 (37.6ºC ± 0.19) and P3 (37.0ºC ± 0.21) varied significantly, independent of the laparoscopy. These data indicated that laparoscopy didn't cause any significant alterations in the...

A laparoscopia ainda é pouco utilizada como ferramenta para técnicas de reprodução assistida em cervídeos sul-americanos, não havendo informações sobre seus efeitos e protocolos anestésicos seguros para sua realização. Objetivaramse avaliar as possíveis alterações na freqüência cardíaca (FC), respiratória (FR), saturação de oxihemoglobina (SpO2) e temperatura retal (TR) durante a laparoscopia para visualização dos órgãos reprodutivos de seis fêmeas de veado-catingueiro (Mazama gouazoubira) anestesiadas com a associação cetamina (5mg/kg), xilazina (0,3mg/kg), midazolam (0,5mg/kg) e isofluorano. Cada animal, após anestesiado, foi posicionado em decúbito dorsal para realização de duas laparoscopias com insuflação abdominal de CO2 (14,2 ± 2,39mmHg; M ± EPM) com intervalo de 40 dias. Para avaliar os principais eventos da laparoscopia, esta foi dividida em três períodos: animal sem insuflação abdominal (P1), com insuflação abdominal (P2) e insuflação abdominal com os quadris elevados a 45º (P3). O controle foi realizado após 40 dias da última laparoscopia, para isto, cada animal foi novamente anestesiado e mantido em decúbito dorsal por um período de tempo igual ao tempo médio de duração das anestesias realizadas nas laparoscopias, sem que o procedimento laparoscópico fosse realizado. O tempo de anestesia dos controles foi também dividido em P1, P2 e P3, respeitando o tempo médio de duração de cada um destes períodos das laparoscopias. Para análise dos dados foi usado o teste de análise de variância (ANOVA) seguido do teste de Tukey e valores de P<0,05 considerados significativos. Não houve diferença significativa nos parâmetros estudados em nenhum dos períodos estabelecidos para o controle e laparoscopia. Porém, a FR média entre P1 (38,8 ± 4,42) e P3 (32,7 ± 4,81) e a TR média entre P1 (38,2ºC ± 0,17), P2 (37,6ºC ± 0,19) e P3 (37,0ºC ± 0,21) variaram significativamente, independente da laparoscopia. Tais dados permitiram concluir que a laparoscopia...