Malformaciones müllerianas: actualización y revisión a propósito de casos clínicos, 2022 / Müllerian malformations: update and review of clinical cases, 2022

Las malformaciones müllerianas (MM) son un grupo de anomalías estructurales originadas por fallas de desarrollo de los conductos paramesonéfricos o de Müller durante las primeras 16 semanas de gestación. Un oportuno diagnóstico y una correcta clasificación permiten ofrecer el mejor manejo y diferenciar aquellas pacientes que requieren tratamiento quirúrgico. Se realizó una revisión de la literatura sobre MM en las bases de datos Epistemonikos, SciELO, Cochrane y PubMed. Se rescataron todas las pacientes ingresadas con diagnóstico de MM. En el año 2021, la American Society of Reproductive Medicine publicó un consenso en el que se estandarizó la nomenclatura, se amplió el espectro y se simplificó la clasificación. La clínica es variada, e incluye pacientes asintomáticas cuyo diagnóstico es un hallazgo por imágenes. Los mejores estudios imagenológicos son la resonancia magnética (RM) y la ultrasonografía 3D, dejando la histeroscopia y la laparoscopia (método de referencia) como procedimiento diagnóstico-terapéutico. Se presentan casos clínicos desarrollados durante el primer trimestre de 2022. Recomendamos la utilización sistemática de la RM para el diagnóstico de anomalías complejas u obstructivas del aparato genital. El tratamiento de estas patologías debe ser realizado por ginecólogos endoscopistas expertos, e incluye tratamiento médico y quirúrgico, el cual debe ser enfocado en cada paciente dependiendo del tipo de MM y de los deseos de fertilidad.

Müllerian malformations (MM) are a group of structural anomalies caused by developmental failure of the paramesonephric or Müllerian ducts during the first 16 weeks of gestation. Timely diagnosis and classification allow us to offer the best management and to differentiate those patients who require surgical treatment. Literature review on MM in Epistemonikos, SciELO, Cochrane and PubMed databases. All patients admitted with a diagnosis of MM were rescued. In 2021, the American Society of Reproductive Medicine publishes a consensus where it standardizes the nomenclature, broadens the spectrum, and simplifies the classification. The clinical picture is varied and includes asymptomatic patients whose diagnosis is an imaging finding. The best imaging studies are magnetic resonance imaging (MRI) and 3D ultrasonography, leaving hysteroscopy and laparoscopy (gold standard) as diagnostic therapeutic. Clinical cases developed during the first trimester 2022 are presented. We recommend the routine use of MRI for the diagnosis of complex and/or obstructive anomalies of the genital tract. The treatment of these pathologies should be performed by expert endoscopic gynecologists and include medical and surgical treatment, which should be focused on each patient, depending on the type of MM and fertility desires.

Detección molecular de la infección natural por parásitos tripanosomátidos en Didelphis marsupialis de una zona rural del norte de Colombia / Molecular detection of the natural infection by trypanosomatid parasites in Didelphis marsupialis from a rural area in northern Colombia

Se evaluó la prevalencia de infección por parásitos tripanosomátidos en Didelphis marsupialis y su relación con los aspectos morfológicos/etarios en una zona rural de El Carmen de Bolívar, Colombia. En cinco visitas (2018-2019) se instalaron trampas Tomahawk® en los ecótopos peridoméstico y silvestre en la Vereda El Alférez, durante tres noches consecutivas/visita. A los animales recolectados, se les determinaron medidas corporales, sexo y edad; y se les extrajo sangre por cardiopuntura, previa sedación, para extracción del ácido desoxirribonucleico (ADN) total y amplificación de la región conservada del ADN de minicírculos de kinetoplasto (ADNk) de parásitos tripanosomátidos. La asociación entre parámetros morfológicos de los didélfidos y su frecuencia de infección por parásitos tripanosomátidos fue determinada mediante una regresión binomial. Se recolectaron 30 individuos de D. marsupialis (60,0% hembras y 40,0% machos/66,7% adultos y 33,3% juveniles). El diagnóstico molecular reveló una frecuencia de infección por parásitos tripanosomátidos del 46,7%. El estadio (p=0,024) fue determinante para la infección. Se discute el papel de D. marsupialis como potencial reservorio de parásitos tripanosomátidos en la zona evaluada.

We studied the prevalence of infection by trypanosomatid parasites in Didelphis marsupialis and its relationship with morphological/age aspects in a rural area of El Carmen de Bolivar, Colombia. Five visits were made to the Vereda El Alférez; each of which lasted three consecutive nights. During these visits, Tomahawk® traps were installed in the peridomestic and wild ecotopes of the Vereda El Alférez. Body measurements, sex and age were determined from the collected animals. Blood was extracted by cardiopuncture, after sedation, in order to obtain total deoxyribonucleic acid (DNA) and amplify the conserved region of the kinetoplast minicircle DNA (kDNA) of parasitic trypanosomatids. The association between morphological parameters of didelphids and their frequency of infection by parasitic trypanosomatids was determined by binomial regression. Thirty D. marsupialis specimens (60.0% females and 40.0% males/66.7% adults and 33.3% juveniles) were collected. Molecular diagnosis revealed a frequency of trypanosomatid parasite infection of 46.7%. Stage (p=0.024) was a determinant for infection. We discuss the role of D. marsupialis as a potential reservoir of parasitic trypanosomatids in the Vereda El Alférez.

Breeding and reproductive behaviour of the neo-tropical opossum, Didelphis marsupialis insularis, Allen 1902 under captive conditions / Reprodução e comportamento reprodutivo do gambá neotropical, Didelphis marsupialis insularis, Allen 1902, em condições de cativeiro

Developing a captive breeding system for the widely hunted Caribbean species of opossum Didelphis marsupialis insularis can greatly aide in the management and conservation of this species in the neo-tropics. Although this species possesses ideal traits for captive breeding in this region (tolerance to high heat and humidity, high reproductive rate, and resistance to disease), challenges due to its aggressive behaviour and limited information on its breeding behaviour have prevented a system from being developed for this species. The present study describes a breeding system, and the reproductive behaviour of this species under captive conditions. Six (1 male; 5 females) adult opossums were maintained and managed for breeding over a ten (10) month period. Pouch litter sizes averaged 5±2.5 with a range of 2 to 8. Gestation length was found to be 13.25±0.96 days and 4 litters (n=23) were successfully weaned at 11-13 weeks. It was found that the male D. m. insularis exhibits behaviours of interest that can serve as indicators for receptivity of the female, and overall, that this species can be successfully reared and bred under captive conditions in the neo-tropics.

O desenvolvimento de um sistema de criação em cativeiro para as espécies de gambá do Caribe amplamente caçadas, Didelphis marsupialis insularis, pode auxiliar bastante no manejo e conservação dessa espécie nos neotrópicos. Embora essa espécie possua características ideais para a criação em cativeiro nessa região (tolerância a altas temperaturas e umidade, alta taxa de reprodução e resistência a doenças), alguns fatores, como o seu comportamento agressivo e informações limitadas sobre seu comportamento reprodutivo, impediram o desenvolvimento de um sistema para D. m. insularis. O presente estudo descreve um sistema de criação e o comportamento reprodutivo dessa espécie em condições de cativeiro. Seis gambás adultos (1 macho e 5 fêmeas) foram mantidos para reprodução durante um período de 10 meses. O tamanho da ninhada em bolsa foi em média de 5 ± 2,5 com um intervalo de 2 a 8. A duração da gestação foi de 13,25 ± 0,96 dias, e 4 ninhadas (n = 23) foram desmamadas com sucesso de 11 a 13 semanas. Verificou-se que o macho D. m. insularis exibe comportamentos de interesse que podem servir como indicadores de receptividade da fêmea, e, em geral, essa espécie pode ser criada com sucesso em condições de cativeiro nos neotrópicos.

Breeding and reproductive behaviour of the neo-tropical opossum, Didelphis marsupialis insularis, Allen 1902 under captive conditions

Abstract Developing a captive breeding system for the widely hunted Caribbean species of opossum Didelphis marsupialis insularis can greatly aide in the management and conservation of this species in the neo-tropics. Although this species possesses ideal traits for captive breeding in this region (tolerance to high heat and humidity, high reproductive rate, and resistance to disease), challenges due to its aggressive behaviour and limited information on its breeding behaviour have prevented a system from being developed for this species. The present study describes a breeding system, and the reproductive behaviour of this species under captive conditions. Six (1 male; 5 females) adult opossums were maintained and managed for breeding over a ten (10) month period. Pouch litter sizes averaged 5±2.5 with a range of 2 to 8. Gestation length was found to be 13.25±0.96 days and 4 litters (n=23) were successfully weaned at 11-13 weeks. It was found that the male D. m. insularis exhibits behaviours of interest that can serve as indicators for receptivity of the female, and overall, that this species can be successfully reared and bred under captive conditions in the neo-tropics.

Resumo O desenvolvimento de um sistema de criação em cativeiro para as espécies de gambá do Caribe amplamente caçadas, Didelphis marsupialis insularis, pode auxiliar bastante no manejo e conservação dessa espécie nos neotrópicos. Embora essa espécie possua características ideais para a criação em cativeiro nessa região (tolerância a altas temperaturas e umidade, alta taxa de reprodução e resistência a doenças), alguns fatores, como o seu comportamento agressivo e informações limitadas sobre seu comportamento reprodutivo, impediram o desenvolvimento de um sistema para D. m. insularis. O presente estudo descreve um sistema de criação e o comportamento reprodutivo dessa espécie em condições de cativeiro. Seis gambás adultos (1 macho e 5 fêmeas) foram mantidos para reprodução durante um período de 10 meses. O tamanho da ninhada em bolsa foi em média de 5 ± 2,5 com um intervalo de 2 a 8. A duração da gestação foi de 13,25 ± 0,96 dias, e 4 ninhadas (n = 23) foram desmamadas com sucesso de 11 a 13 semanas. Verificou-se que o macho D. m. insularis exibe comportamentos de interesse que podem servir como indicadores de receptividade da fêmea, e, em geral, essa espécie pode ser criada com sucesso em condições de cativeiro nos neotrópicos.

Breeding and reproductive behaviour of the neo-tropical opossum, Didelphis marsupialis insularis, Allen 1902 under captive conditions / Reprodução e comportamento reprodutivo do gambá neotropical, Didelphis marsupialis insularis, Allen 1902, em condições de cativeiro

Developing a captive breeding system for the widely hunted Caribbean species of opossum Didelphis marsupialis insularis can greatly aide in the management and conservation of this species in the neo-tropics. Although this species possesses ideal traits for captive breeding in this region (tolerance to high heat and humidity, high reproductive rate, and resistance to disease), challenges due to its aggressive behaviour and limited information on its breeding behaviour have prevented a system from being developed for this species. The present study describes a breeding system, and the reproductive behaviour of this species under captive conditions. Six (1 male; 5 females) adult opossums were maintained and managed for breeding over a ten (10) month period. Pouch litter sizes averaged 5±2.5 with a range of 2 to 8. Gestation length was found to be 13.25±0.96 days and 4 litters (n=23) were successfully weaned at 11-13 weeks. It was found that the male D. m. insularis exhibits behaviours of interest that can serve as indicators for receptivity of the female, and overall, that this species can be successfully reared and bred under captive conditions in the neo-tropics.

O desenvolvimento de um sistema de criação em cativeiro para as espécies de gambá do Caribe amplamente caçadas, Didelphis marsupialis insularis, pode auxiliar bastante no manejo e conservação dessa espécie nos neotrópicos. Embora essa espécie possua características ideais para a criação em cativeiro nessa região (tolerância a altas temperaturas e umidade, alta taxa de reprodução e resistência a doenças), alguns fatores, como o seu comportamento agressivo e informações limitadas sobre seu comportamento reprodutivo, impediram o desenvolvimento de um sistema para D. m. insularis. O presente estudo descreve um sistema de criação e o comportamento reprodutivo dessa espécie em condições de cativeiro. Seis gambás adultos (1 macho e 5 fêmeas) foram mantidos para reprodução durante um período de 10 meses. O tamanho da ninhada em bolsa foi em média de 5 ± 2,5 com um intervalo de 2 a 8. A duração da gestação foi de 13,25 ± 0,96 dias, e 4 ninhadas (n = 23) foram desmamadas com sucesso de 11 a 13 semanas. Verificou-se que o macho D. m. insularis exibe comportamentos de interesse que podem servir como indicadores de receptividade da fêmea, e, em geral, essa espécie pode ser criada com sucesso em condições de cativeiro nos neotrópicos.

Sarcocystis neurona, seroprevalence of antibodies in equines and research of oocystis in opossum in Ilhéus - Itabuna microregion, Bahia, Brazil / Sarcocystis neurona, seroprevalência de anticorpos em equinos e pesquisa de oocistos em gambás na microrregião de Ilhéus - Itabuna, Bahia, Brasil

Abstract The aims of this study were to determine the seroprevalence of Sarcocystis neurona antibodies in equines in the Ilhéus-Itabuna microregion (BA), and identify possible factors associated with infection. The presence of sporocysts/oocysts of Sarcocystis spp. was also verified in Didelphis spp. A total of 669 serum samples were collected from equines in 56 properties located in 12 municipalities in the region. Indirect fluorescent antibody test was performed with slides containing merozoites of the S. neurona, using a cut-off titer of 1:80. Occurrence of 7.92% of anti-S. neurona antibodies was observed in the sampled equines. The purposes trade and work were significantly associated with the presence of antibodies (p<0.05), and being used for the purpose of work (21.6%) was considered a risk factor, while being used for the purpose of trade (3.6%) was a protective factor. A total of 25 Didelphis spp. was captured for research on sporocysts/oocysts in stool samples and intestinal scrapings, being all negative. Didelphis spp. were all negative for the presence of Sarcocystis spp. and this circumstance does not change the fact that seroprevalence of S. neurona has been observed in horses raised in the southern Bahia.

Resumo O presente estudo foi realizado na microrregião de Ilhéus-Itabuna, Bahia. Os objetivos deste estudo foram determinar a soroprevalência de anticorpos contra Sarcocystis neurona em equinos da microrregião Ilhéus-Itabuna (BA) e identificar possíveis fatores associados à infecção. A presença de esporocistos/oocistos de Sarcocystis spp. também foi pesquisada em Didelphis spp. Foram coletadas 669 amostras de soro de equinos em 56 propriedades localizadas em 12 municípios da região. Foi utilizada a reação de imunofluorescência indireta (RIFI), utilizando-se lâminas confeccionadas com merozoítos de Sarcocystis neurona (cepa SN138) e ponto de corte na diluição de 1:80. A ocorrência de anticorpos anti- S. neurona, nos equinos amostrados, foi de 7,92%. As finalidades dos animais - comércio e trabalho - apresentaram-se significativas (p<0.05), sendo que a finalidade trabalho (21,6%) foi considerada fator de risco, enquanto a finalidade comércio (3,6%) foi considerada fator de proteção. Foram capturados 25 Didelphis spp., para pesquisa de esporocistos/oocistos em amostras de fezes e raspado de mucosa intestinal. Todos os Didelphis spp. foram negativos para a presença de Sarcocystis spp., mesmo assim essa circunstância não alterou o fato da ocorrência de S. neurona ter sido observada em cavalos criados na mesorregião do sul da Bahia.

Rickets in white-eared opossums (Didelphis albiventris) / Raquitismo em gambás-de-orelha-branca (Didelphis albiventris)

Two young opossums were necropsied and diagnosed with rickets. This study aims to describe the clinical-pathological aspects of rickets in Didelphis albiventris. Macroscopically, the opossums presented kyphosis and scoliosis, lateral deviation of the limbs in varus, locomotion difficulty, and enlargement with softening of costochondral junctions (rickety rosary). Samples of bones and joints were processed for hematoxylin and eosin staining and Masson's trichrome. Microscopically, we observed thickening of the epiphyseal plate, characterized by irregular and multifocal proliferation of serialized and hypertrophic cartilage zones, which formed circular groups of large, dysplastic chondrocytes towards the spongy zone, often surrounded by non-mineralized osteoid tissue. In the cortical bone, there were pale eosinophilic zones around the Havers channels consistent with non-mineralized osteoid. The staining of Masson's trichrome evidenced the accumulation of osteoid tissue in cortical and trabecular bones. It is possible that a mixed cause of absorption deficiency of vitamin D3 associated with an unbalanced Ca:P diet based on lactose-free milk and fruits may have triggered the disease.(AU)

Dois gambás jovens foram necropsiados e diagnosticados com raquitismo. O objetivo do trabalho é descrever os aspectos clínico-patológicos de raquitismo em Didelphis albiventris. Macroscopicamente os gambás apresentaram cifose e escoliose, desvio lateral dos membros em varus, dificuldade de locomoção e alargamento com amolecimento das junções costocondrais (rosário raquítico). Amostras dos ossos e articulações foram processadas para coloração de hematoxilina e eosina e Tricrômico de Masson. Microscopicamente havia espessamento da placa epifisária, caracterizada pela proliferação irregular e multifocal das zonas de cartilagem seriada e hipertrófica, que formavam grupos circulares de condrócitos grandes, displásicos em direção a zona esponjosa frequentemente cercados por tecido osteoide não mineralizado. No osso cortical haviam zonas eosinofílicas pálidas ao redor dos canais de Havers consistentes com osteoide não mineralizado. A coloração de Tricrômico de Masson evidenciou o acúmulo de tecido osteoide no nosso cortical e trabecular. Acredita-se que uma causa mista de déficit de absorção de vitamina D3 associada a uma dieta desbalanceada em Ca:P a base de leite sem lactose e frutas tenha desencadeado a doença.(AU)

A case of polymelia associated with syndactyly in Didelphis aurita (Wied-Neuwied, 1826) / Relato de caso de polimelia associada a sindactilia em Didelphis aurita (Wied-Neuwied, 1826)

Abstract In January 2018, a young male of Didelphis aurita (Wied-Neuwied, 1826), the Brazilian common opossum, was admitted for veterinary evaluation at the Recovery Clinic of Wild Animals of Estácio de Sá University, CRAS, in Rio de Janeiro, Brazil, due to the presence of a partially duplicated right hind limb. After x-ray evaluation, polymelia associated with syndactyly was diagnosed. The images revealed the following alterations on the right limb: single femur with increased diameter, two tibiae, two fibulae and absence of hallux. The observed malformations were restricted to the hind right limb, which remains functional. This is the first report of such anomalies in an individual of Didelphis aurita.

Resumo Em janeiro de 2018, um macho jovem de Didelphis aurita (Wied-Neuwied, 1826), o gambá-de-orelha-preta, foi recebido para atendimento veterinário na Clínica de Recuperação de Animais Silvestres (CRAS) da Universidade Estácio de Sá, no Rio de Janeiro, por ser portador de membro posterior direito parcialmente duplicado. Após exame radiográfico, constatou-se a ocorrência de polimelia associada a sindactilia. As imagens radiográficas revelaram as seguintes alterações no membro posterior direito: um fêmur com diâmetro aumentado, duas tíbias, duas fíbulas e ausência de hálux. As malformações observadas restringiram-se apenas ao membro posterior direito, o qual mantém sua funcionalidade. Essas alterações nunca antes foram descritas na espécie Didelphis aurita.

Wildlife rehabilitation: A case study of the neo-tropical, opossum Didelphis marsupialis insularis, Allen 1902 / Reabilitação de vida selvagem: um estudo de caso do gambá neotropical, Didelphis marsupialis insularis, Allen 1902

Abstract Wildlife rehabilitation facilities in the Caribbean region are limited, yet they can provide relevant information on wild populations. Didelphis marsupialis insularis is a popularly hunted, under-studied, neo-tropical marsupial species that is increasingly being admitted for rehabilitation. The aim of this study was 1. To record the experiences of rehabilitating D. marsupialis insularis in the neo-tropical island of Trinidad and Tobago and 2. To extract and highlight information on the biology of this opossum sub-species. Using admission records, obtained over a roughly four year period, two breeding periods (February to March and August to October) were illustrated. Litter sizes averaged five individuals, with a range of 1 to 8 young. This species was found to be common in urban areas of the country, with dog attacks reported as the major cause for admission. Thus the information recorded by this wildlife rehabilitation facility has provided great insight on the sparsely studied opossum, D. marsupialis insularis.

Resumo As instalações de reabilitação e assistência à vida selvagem na região neotropical do Caribe são limitadas, mas informações relevantes sobre populações selvagens podem ser obtidas nesses locais. Didelphis marsupialis insularis é uma espécie de marsupial neotropical, subestimada e popularmente caçada; no entanto, essa espécie está sendo cada vez mais admitida para a reabilitação. Os objetivos deste estudo foram: 1. Registrar as experiências de reabilitação de D. marsupialis insularis na ilha neotropical de Trinidad e Tobago; e 2. Extrair e destacar informações sobre a biologia dessa subespécie de gambá. Utilizando registros de admissão, obtidos ao longo de um período de aproximadamente quatro anos, foram mostrados dois períodos de reprodução, de fevereiro a março e de agosto a outubro. O tamanho da ninhada foi em média de cinco indivíduos, variando de um a oito jovens. Comumente encontrado em áreas urbanas do país, os ataques de cães foram relatados como a principal causa de internação dessa espécie. Assim, a informação registrada por essa instalação de reabilitação de vida selvagem forneceu uma grande visão sobre o gambá D. marsupialis insularis, que tem sido pouco estudado.

Common opossum population density in an agroforestry system in Bolivia

The common opossum, Didelphis marsupialis thrives near human settlements. Understanding its ecology could help planning management decisions about this species, especially in anthropogenic landscapes. Yet, there are no density estimations for this species throughout its distribution range in Bolivia. We estimated the density of D. marsupialis in a rural agricultural community, where agroforestry plantations and fallows cover most of the land. We counted individuals in line transects and used DISTANCE software to calculate density. We covered a total of 70.21 km in 143 night counts, obtaining 38 records of D. marsupialis. We estimated a density of 0.30 individuals ha-1 (SE = 0.062; range: 0.20 - 0.45 individuals ha-1), with a mean encounter rate of 0.54 individuals km-1. Encounter rate varied between habitats, with mean values of 0.20 in secondary forests and 0.64 in agroforestry plantations. Our density estimate is near the lower range of previously reported values for the common opossum in other countries. We argue that our results may reflect the response to the availability of food resources and predation pressure in agroforestry plantations. (AU)

Fibroblastic osteosarcoma in a white-eared opossum (Didelphis albiventris) / Osteossarcoma fibroblástico em gambá-de-orelha-branca (Didelphis albiventris)

Osteosarcoma is characterized by the production of osteoid or woven bone, using neoplastic osteoblasts. It is the most common primary bone neoplasm in canines and humans. This neoplasm was previously reported in all vertebrate classes, including a wide variety of mammals. However, there is no case report describing this neoplasm in Didelphis albiventris. Therefore, the objective of this manuscript is to describe the clinical-pathological aspects of fibroblastic osteosarcoma in D. albiventris. A wild adult male white-eared opossum (D. albiventris) arrived at the zoological park of the "Universidade de Caxias do Sul" with a swollen left thoracic limb. After a general clinical examination, the animal was transferred to the Veterinary Clinic of "Universidade de Caxias do Sul" for radiographic examination of the left thoracic limb and thorax. Additionally, some material was collected through fine needle aspiration (FNA) for cytologic evaluation. The radiographic findings and cytologic evaluation indicated osteosarcoma. The animal was euthanized due to severe clinical conditions and guarded prognosis. During necropsy, macroscopic analysis of the viscera was performed, fragments of various organs were collected and fixed in 10% neutral buffered formalin. All fragments were processed following routine histological techniques. The histopathological evaluation confirmed osteosarcoma, which was classified as a fibroblastic subtype. Case reports are crucial for the knowledge of incidence, prevalence, and behavior of the current mentioned disease, as well as other diseases, in species with such limited information. In order to obtain a decisive diagnosis, a few different examination methods were associated. Although the observations presented are based on a single case, this neoplasm had a similar clinical presentation to that described in other species.(AU)

O osteossarcoma é caracterizado pela produção de osteoide ou osso imaturo, por osteoblastos neoplásicos. É a neoplasia óssea primária mais comum em caninos e humanos. Essa neoplasia já foi relatada em todas as classes de vertebrados, incluindo uma grande variedade de mamíferos. Não havendo descrição dessa neoplasia até o momento em Didelphis albiventris. O objetivo deste trabalho é descrever aspectos clínico-patológicos de um caso de osteossarcoma fibroblástico em D. albiventris. Chegou para atendimento no Zoológico da Universidade de Caxias do Sul um gambá-de-orelha-branca (D. albiventris), macho, adulto, de vida livre com aumento de volume no membro torácico esquerdo. Após avaliação clínica geral, o animal foi encaminhado para a Clínica Veterinária da Universidade de Caxias do Sul para realização de radiografia do membro torácico esquerdo e de tórax, sendo também realizada coleta de material por punção aspirativa por agulha fina (PAAF) para avaliação citológica. Os achados radiográficos e da avaliação citológica foram sugestivos de osteossarcoma. Devido ao estado clínico grave e prognóstico reservado optou-se pela eutanásia. Durante a necropsia realizou-se a análise macroscópica das vísceras, foram coletados fragmentos de diversos órgãos, fixados em formalina 10%, processados pelas técnicas histológicas de rotina. Na avaliação histopatológica confirmou-se a suspeita de osteossarcoma sendo classificado no subtipo fibroblástico. A descrição de relatos é fundamental para conhecimento da incidência, prevalência e comportamento desta e de outras doenças em espécies que as informações são limitadas. A associação de diferentes métodos de exames foram necessários para a obtenção de um diagnóstico definitivo. Embora as observações apresentadas se baseiem em um caso único, esta neoplasia possuiu apresentação clínica semelhante a descrita em outras espécies.(AU)

Rickettsial infection in equids, opossums and ticks in the municipality of Monte Mor, state of São Paulo, Brazil / Infecção por Rickettsia spp. em equídeos, gambás e carrapatos do município de Monte Mor, estado de São Paulo, Brasil

Abstract The aim of this study was to investigate rickettsial infection in equids, opossums and ticks in the municipality of Monte Mor, a place where a Brazilian spotted fever case occurred in 2005. In addition, characteristics possibly associated with seropositivity in horses were analyzed. Serum samples from horses, mules and opossums (Didelphis albiventris) were subjected to indirect immunofluorescence assay (IFA) against Rickettsia rickettsii. The ticks collected from the animals were identified and Amblyomma sculptum ticks from the equids were tested using PCR for Rickettsia spp. Anti-R. rickettsii antibodies were detected in 22.6% (14/62) of the horses, none of the mules and 21.7% (5/23) of the opossums. Among the variables analyzed, only age > 12 years showed a statistically significant association with seropositivity among horses. All of the 166 A. sculptum ticks tested using PCR were negative. The results showed that rickettsiae of the spotted fever group was circulating in the municipality of Monte Mor when the samples were collected and indicate a need for surveillance of Brazilian spotted fever in this region.

Resumo Este trabalho objetivou pesquisar a infecção por Rickettsia spp. em equídeos, gambás e carrapatos, do município de Monte Mor, local que teve um caso de febre maculosa brasileira, em 2005. Além disso, características possivelmente associadas com a soropositividade nos equinos foram analisadas. Soros de equinos, muares e gambás Didelphis albiventris foram submetidos à reação de imunofluorescência indireta (RIFI) contra Rickettsia rickettsii. Os carrapatos coletados dos animais foram identificados e os carrapatos Amblyomma sculptum dos equídeos foram testados pela PCR para Rickettsia spp. Anticorpos anti-R. rickettsii foram detectados em 22,6% (14/62) equinos, zero muares e 21,7% (5/23) gambás. Entre as variáveis analisadas, apenas a idade maior que 12 anos mostrou associação estatisticamente significante com a soropositividade em equinos. De 166 carrapatos A. sculptum testados pela PCR, todos foram negativos. Os resultados mostram que riquétsias do grupo da febre maculosa estavam circulando no município de Monte Mor, quando as amostras foram coletadas, e apontam para a necessidade de vigilância para a febre maculosa brasileira nessa região.

Molecular isolation and identification of Mycobacterium avium subsp. hominissuis in Didelphis virginiana from Hidalgo, Mexico

Objective: To isolate and identify the exact species of the genus Mycobacterium from Didelphis (D.) virginiana, and the direct implications of this bacterium to public health and veterinary medicine. Methods: Thirty-one D. virginiana were captured and necropsied in Hidalgo, Mexico. Tissue samples were collected to culture mycobacteria present and examine individual specimens' histopathology. Mycobacterium identification was obtained through the application of amplification and sequencing of 16S rDNA techniques. Results: Three strains were isolated and identified as Mycobacterium (M.) avium subsp. hominissuis by utilizing M. avium complex- specific primers. Granulomatous lesions were observed in the subpleural zone (granuloma grade Π ) and bronchial (granuloma grade I ) of the lungs of D. virginiana with positive isolation. Conclusions: Three strains of M. avium subsp. hominissuis, from lung tissue samples of D. virginiana were identified. This subspecies of M. avium has important implications in public health and veterinary medicine.

Molecular isolation and identification of Mycobacterium avium subsp. hominissuis in Didelphis virginiana from Hidalgo, Mexico

Objective: To isolate and identify the exact species of the genus Mycobacterium from Didelphis (D.) virginiana, and the direct implications of this bacterium to public health and veterinary medicine. Methods: Thirty-one D. virginiana were captured and necropsied in Hidalgo, Mexico. Tissue samples were collected to culture mycobacteria present and examine individual specimens' histopathology. Mycobacterium identification was obtained through the application of amplification and sequencing of 16S rDNA techniques. Results: Three strains were isolated and identified as Mycobacterium (M.) avium subsp. hominissuis by utilizing M. avium complex- specific primers. Granulomatous lesions were observed in the subpleural zone (granuloma grade Π ) and bronchial (granuloma grade I ) of the lungs of D. virginiana with positive isolation. Conclusions: Three strains of M. avium subsp. hominissuis, from lung tissue samples of D. virginiana were identified. This subspecies of M. avium has important implications in public health and veterinary medicine.

Surveillance of phlebotomine fauna and Didelphis marsupialis (Didelphimorphia: Didelphidae) infection in an area highly endemic for visceral leishmaniasis in Colombia / Vigilancia de la fauna flebotomínea e infección de Didelphis marsupialis (Didelphimorphia: Didelphidae) en un área de alta endemia de leishmaniasis visceral en Colombia

Abstract Introduction: The study of the interaction between the parasite, the vector and the mammalian hosts, including man, allows to understand the behavior of the leishmaniases. Objective: To determine the presence of Lutzomyia species and to detect the Leishmania infection in Didelphis marsupialis in an endemic area for visceral leishmaniasis. Materials and methods: Phlebotomine fauna and individuals of D. marsupialis were collected with CDC and Tomahawk™ traps, respectively. The species of Lutzomyia were identified using the Young and Duncan key (1994). Ear and tail biopsies and blood samples from D. marsupialis were taken to identify the Leishmania species by amplifying a fragment of the gene associated with the 70 kD heat shock protein. Results: Seven Lutzomyia species were identified: Lu. evansi, Lu. gomezi, Lu. panamensis, Lu. dubitans, Lu. cayennensis cayennensis, Lu. rangeliana and Lu. trinidadensis. The first three species have epidemiological importance in Colombia because of their implications in the transmission of the Leishmania parasite. Sixty-five tissue samples from 19 D. marsupialis individuals were negative for Leishmania spp. Conclusions: The presence of the Lutzomyia species that have been identified as vectors for Leishmania inside and around houses in the village of El Bledo, in El Carmen de Bolívar represents a risk of infection. Furthermore, the presence of Lu. panamensis is reported for first time in El Carmen de Bolívar in Colombia. Although the lack of detection of Leishmania spp. in D. marsupialis samples may suggest that D. marsupialis does not play an important role in the transmission cycle of Leishmania in this region, it is necessary to carry out further longitudinal studies to confirm this hypothesis.

Resumen Introducción. El estudio de la interacción entre el parásito, el vector y los huéspedes mamíferos, incluido el hombre, permite entender el comportamiento de la leishmaniasis. Objetivo. Determinar la presencia de especies del género Lutzomyia y detectar la infección por Leishmania spp. en Didelphis marsupialis en un área endémica de leishmaniasis visceral. Materiales y métodos. Se recolectaron flebotomíneos y D. marsupialis con trampas CDC y Tomahawk™, respectivamente. Las especies de Lutzomyia se identificaron usando la clave de Young y Duncan, 1994. Se tomaron biopsias de oreja, cola y muestras de sangre de D. marsupialis para diagnosticar Leishmania spp. mediante la amplificación de un fragmento del gen de la proteína de choque térmico de 70 kD. Resultados. Se identificaron siete especies de Lutzomyia: Lu. evansi, Lu. gomezi, Lu. panamensis, Lu. dubitans, Lu. cayennensis cayennensis, Lu. rangeliana y Lu. trinidadensis. Las tres primeras especies son reconocidas como vectores en el país por estar implicadas en la transmisión de Leishmania spp. En total, 65 muestras de tejidos de oreja, cola y de sangre provenientes de 19 D. marsupialis fueron negativas para Leishmania spp. en la PCR-HSP70. Conclusiones. La presencia de flebotomíneos con importancia epidemiológica en la zona evaluada representa un riesgo de transmisión. Asimismo, Lu. panamensis es reportada por primera vez en El Bledo (Carmen de Bolívar). La ausencia de Leishmania spp. en D. marsupialis podría sugerir que esta especie no tiene un papel importante en el ciclo de transmisión de Leishmania en la vereda El Bledo, por lo que es necesario profundizar en estudios longitudinales para corroborar esta hipótesis.

Detection of a putative novel genotype of Ehrlichia sp. from opossums (Didelphis aurita) from Brazil / Detecção de um suposto novo genótipo de Ehrlichia sp. em gambás (Didelphis aurita) do Brasil

Abstract Erlichiosis affects humans and animals worldwide. Its distribution and prevalence depends on the presence of tick vectors and hosts in one geographic area. The aim of the present study was to investigate the occurrence of Ehrlichia spp. and Anaplasma spp. in opossums (Didelphis sp.) from the State of Rio de Janeiro, southeast Brazil. Blood samples from 37 animals were tested for these two pathogens using molecular methods. One animal (2.7%) was positive for Ehrlichia sp. by 16S rRNA-based nested PCR. In a phylogenetic analysis based on the 16S rRNA gene using the maximum likelihood method and the GTRGAMMA+I evolutionary model, we detected a novel Ehrlichia sp. genotype closely related to genotypes of E. canis previously reported in dogs from Brazil. To the authors' knowledge, this is the first molecular detection of Ehrlichia sp. in opossums from this State in the southeastern region of the country.

Resumo A erliquiose afeta seres humanos e animais em todo o mundo. Sua distribuição e prevalência dependem da presença de vetores de carrapatos e hospedeiros em uma área geográfica. O objetivo do presente estudo foi investigar a ocorrência de Ehrlichia sp. e Anaplasma sp. em gambás (Didelphis sp.) do Estado do Rio de Janeiro, sudeste do Brasil. Amostras de sangue de 37 animais foram testadas para estes dois patógenos usando métodos moleculares. Um animal (2,7%) foi positivo para Ehrlichia sp. baseado em 16S rRNA-nested PCR. Em uma análise filogenética baseada no gene 16S rRNA usando o método de máxima verossimilhança e o modelo evolutivo GTRGAMMA + I, detectamos um novo genótipo de Ehrlichia sp. intimamente relacionado a genótipos de E. canis previamente relatados em cães do Brasil. Para o conhecimento dos autores, esta é a primeira detecção molecular de Ehrlichia sp. em gambás deste estado na região sudeste do país.

Trypanosoma janseni n. sp. (Trypanosomatida: Trypanosomatidae) isolated from Didelphis aurita (Mammalia: Didelphidae) in the Atlantic Rainforest of Rio de Janeiro, Brazil: integrative taxonomy and phylogeography within the Trypanosoma cruzi clade

BACKGROUND Didelphis spp. are a South American marsupial species that are among the most ancient hosts for the Trypanosoma spp. OBJECTIVES We characterise a new species (Trypanosoma janseni n. sp.) isolated from the spleen and liver tissues of Didelphis aurita in the Atlantic Rainforest of Rio de Janeiro, Brazil. METHODS The parasites were isolated and a growth curve was performed in NNN and Schneider's media containing 10% foetal bovine serum. Parasite morphology was evaluated via light microscopy on Giemsa-stained culture smears, as well as scanning and transmission electron microscopy. Molecular taxonomy was based on a partial region (737-bp) of the small subunit (18S) ribosomal RNA gene and 708 bp of the nuclear marker, glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) genes. Maximum likelihood and Bayesian inference methods were used to perform a species coalescent analysis and to generate individual and concatenated gene trees. Divergence times among species that belong to the T. cruzi clade were also inferred. FINDINGS In vitro growth curves demonstrated a very short log phase, achieving a maximum growth rate at day 3 followed by a sharp decline. Only epimastigote forms were observed under light and scanning microscopy. Transmission electron microscopy analysis showed structures typical to Trypanosoma spp., except one structure that presented as single-membraned, usually grouped in stacks of three or four. Phylogeography analyses confirmed the distinct species status of T. janseni n. sp. within the T. cruzi clade. Trypanosoma janseni n. sp. clusters with T. wauwau in a well-supported clade, which is exclusive and monophyletic. The separation of the South American T. wauwau + T. janseni coincides with the separation of the Southern Super Continent. CONCLUSIONS This clade is a sister group of the trypanosomes found in Australian marsupials and its discovery sheds light on the initial diversification process based on what we currently know about the T. cruzi clade.

Helmintofauna de Didelphis albiventris (lund, 1841) no município de Palotina, Paraná, Brasil / Didelphis albiventris (lund, 1841) helminths in the city of Palotina, Paraná, Brazil / Helmintofauna de Didelphis albiventris (lund, 1841) en el municipio de Palotina, Parana, Brasil

Entre fevereiro de 2009 a abril de 2011, nove exemplares adultos (sete machos e duas fêmeas) de Didelphis albiventris (Lund, 1841), foram encontrados mortos por atropelamento no município de Palotina, Paraná. Os animais foram encaminhados para necropsia e coleta de material. Os parasitos encontrados foram fixados em formol acético e, posteriormente identificados. Os nove gambás estavam parasitados por helmintos, com predominância para o filo Nemathelminthes (66%), seguido por Acanthocephala (17%) e Platyhelminthes (17%). Foram isolados os seguintes helmintos e respectivas prevalências: Turgida turgida (89%) no estômago; Cruzia tentaculata (44%), Aspidodera sp. (22%) e Trichuris sp. (11%) no intestino grosso; Hamanniella microcephala (78%) e Rhopalias coronatus (11%) no intestino delgado. Em 80% dos D. albiventris ocorreu a associação parasitária entre, no mínimo, duas espécies de helmintos.(AU)

From February 2009 to April 2011, nine adult specimens (seven male and two female) of Didelphis albiventris (Lund, 1841) were found dead by roadkill in Palotina, Paraná. The animals were submitted for necropsy and collection of material. The parasites found were fixed in formalin acetic acid and subsequently identified. The nine opossums were parasitized by helminths, predominantly belonging to the Nemathelminthes (66%) phylum, followed by Acanthocephala (17%) and Platyhelminthes (17%). The following helminths were isolated and their prevalences were as follows: Turgida turgida (89%) in the stomach; Cruzia tentaculata (44%), Aspidodera sp. (22%) and Trichuris sp. (11%) in the large intestine; Hamanniella microcephala (78%) and Rhopalias coronatus (11%) in the small intestine.Parasitic association between at least two helminth species occurred in 80% of D. albiventris samples.(AU)

De febrero de 2009 a abril de 2011, nueve ejemplares adultos (siete machos y dos hembras) de Didelphis albiventris (Lund, 1841), fueron encontrados muertos por arrollamiento en el municipio de Palotina, Paraná. Los animales fueron enviados para necropsia y colecta de material. Los parásitos encontrados fueron fijados en formol acético, y posteriormente identificados. Las nueve zarigüeyas estaban parasitadas por helmintos, especialmente por Nemathelminthes (66%), seguido de Acanthocephala (17%) y Platyhelminthes (17%). Fueron aislados los siguientes helmintos y sus respectivas prevalencias: Turgida turgida (89%) en el estómago; Cruzia tentaculata (44%), Aspidodera sp. (22%) y Trichuris sp. (11%) en el intestino grueso; Hamanniella microcephala (78%) y Rhopalias coronatus (11%) en el intestino delgado. En 80% de los D. albiventris ocurrió la asociación parasitaria entre al menos dos especies de helmintos.(AU)

Riesgo de transmisión de la enfermedad de Chagas por intrusión de triatominos y mamíferos silvestres en Bucaramanga, Santander, Colombia / Risk of transmission of Chagas disease by intrusion of triatomines and wild mammals in Bucaramanga, Santander, Colombia

Introducción: La notificación de triatominos en las viviendas de algunos barrios de Bucaramanga motivó la realización de este estudio. Objetivo: Evaluar la intrusión de triatominos y mamíferos, así como algunos factores de riesgo para la enfermedad de Chagas en viviendas urbanas. Materiales y métodos: En un barrio de Bucaramanga, Santander, se recolectaron triatominos mensualmente durante un año con participación comunitaria mediante búsqueda manual en el alumbrado público, y el uso de trampas de luz, cebo animal y atrayentes químicos en el bosque cercano. Los reservorios se recolectaron con trampas cebadas. Los insectos y mamíferos se determinaron y examinaron para establecer su infección natural. Los factores de riesgo de las viviendas se midieron mediante una encuesta sobre factores sociales y ambientales. Resultados: Se recolectaron 11 adultos de Panstrongylus geniculatus y 63 de Rhodnius pallescens en el bosque, en zonas de recreación en el peridomicilio y en el domicilio, incluidas dos hembras y 21 ninfas de R. pallescens en dormitorios. Se capturaron dos ejemplares de Didelphis marsupialis en el bosques adyacente. De los 11 individuos de P. geniculatus capturados, se examinaron nueve, de los cuales cinco fueron positivos para Trypanosoma cruzi (56 %); de los 63 individuos de R. pallescens capturados, se examinaron ocho, cuatro de los cuales fueron positivos para T. cruzi (50 %). De dos especímenes de D. marsupiales capturados, uno fue examinado y se encontró que era positivo para T. cruzi. No se pudo establecer un factor de riesgo significativo, sin embargo, las viviendas con reporte de triatominos se encontraban más cerca del bosque adyacente. Conclusiones: El hallazgo de especies de triatominos intrusivas y de mamíferos con T. cruzi en el domicilio y el peridomicilio, así como en los bosques periurbanos, demuestra el riesgo de infección en las poblaciones que habitan en viviendas urbanas adyacentes a los ecótopos donde se mantiene el ciclo silvestre.

Abstract Introduction: Notice of triatomines in dwellings of some neighborhoods in Bucaramanga motivated the realization of this study. Objetive: To evaluate the intrusion of triatomines and mammals, as well as some risk factors in urban dwellings. Materials and methods: Triatomines were collected in a neighborhood in Bucaramanga, Santander, on a monthly basis during one year with participation of the community. Collection included manual search in lamp posts, use of light traps, animal bait, and chemical attractants in nearby forests. Reservoirs were collected with bait traps. Insects and mammals were identified and examined in order to determine their natural infection. Risk factors in homes were assessed by means of a social-environmental survey. Results: Eleven adult specimens of Pastrongylus geniculatus, as well as 63 of Rhodnius pallescens were collected in the forest, recreational peridomiciliary areas, and houses. Even two females and 21 nymphs of R. pallescens were found in bedrooms. Two specimens of Didelphis marsupialis were captured in neighboring forests. Out of the eleven P. geniculatus captured, nine were examined. Of these, five were positive for Trypanosoma cruzi. It was not possible to establish a significant risk factor; however, the dwellings with report of triatomines were located nearer to the adjacent forest. Conclusions: The finding of intrusive triatominae species and mammals with T. cruzi in intradomiciliary and peridomiciliary areas and periurban forests prove the potential risk to acquire infection from these populations that dwell in urban housing adjacent to these ecotopes where the sylvan cycle is kept.

Detection of a putative novel genotype of Ehrlichia sp. from opossums (Didelphis aurita) from Brazil

Abstract Erlichiosis affects humans and animals worldwide. Its distribution and prevalence depends on the presence of tick vectors and hosts in one geographic area. The aim of the present study was to investigate the occurrence of Ehrlichia spp. and Anaplasma spp. in opossums (Didelphis sp.) from the State of Rio de Janeiro, southeast Brazil. Blood samples from 37 animals were tested for these two pathogens using molecular methods. One animal (2.7%) was positive for Ehrlichia sp. by 16S rRNA-based nested PCR. In a phylogenetic analysis based on the 16S rRNA gene using the maximum likelihood method and the GTRGAMMA+I evolutionary model, we detected a novel Ehrlichia sp. genotype closely related to genotypes of E. canis previously reported in dogs from Brazil. To the authors knowledge, this is the first molecular detection of Ehrlichia sp. in opossums from this State in the southeastern region of the country.

Resumo A erliquiose afeta seres humanos e animais em todo o mundo. Sua distribuição e prevalência dependem da presença de vetores de carrapatos e hospedeiros em uma área geográfica. O objetivo do presente estudo foi investigar a ocorrência de Ehrlichia sp. e Anaplasma sp. em gambás (Didelphis sp.) do Estado do Rio de Janeiro, sudeste do Brasil. Amostras de sangue de 37 animais foram testadas para estes dois patógenos usando métodos moleculares. Um animal (2,7%) foi positivo para Ehrlichia sp. baseado em 16S rRNA-nested PCR. Em uma análise filogenética baseada no gene 16S rRNA usando o método de máxima verossimilhança e o modelo evolutivo GTRGAMMA + I, detectamos um novo genótipo de Ehrlichia sp. intimamente relacionado a genótipos de E. canis previamente relatados em cães do Brasil. Para o conhecimento dos autores, esta é a primeira detecção molecular de Ehrlichia sp. em gambás deste estado na região sudeste do país.