Molecular detection of hemoplasmas in rescued blackeared opossums (Didelphis aurita Wied-Neuwied, 1826) from southeastern Brazil, with evidence of a novel genotype infecting marsupials / Detecção molecular de hemoplasmas em gambás-de-orelha-preta (Didelphis aurita Wied-Neuwied, 1826) resgatados do sudeste do Brasil, com evidência de um novo genótipo infectando marsupiais

There is a growing concern about the participation of wild hosts and reservoirs in the epidemiology of several pathogens, particularly within the context of environmental changes and the expansion of the One Health concept. The aim of this study was to investigate the presence of hemoplasmas in opossums rescued from the metropolitan region of Rio de Janeiro state, Brazil. Blood samples were collected from 15 Didelphis aurita and subjected to DNA extraction and PCR using primers for the 16S rRNA and 23S rRNA genes. Physical examination and hematological analysis were also performed. Three out of 15 opossums tested positive for hemotropic Mycoplasma spp. by PCR and showed hematological alterations such as anemia and leukocytosis. Clinical signs were non-specific and associated to traumatic lesions. The phylogenetic analysis indicated that the hemoplasma detected was positioned between 'Ca. Mycoplasma haemodidelphis' detected in D. virginiana from North American and hemoplasmas recently detected in D. aurita from the state of Minas Gerais, Brazil. This study indicates the existence of hemoplasma infections in D. aurita from the metropolitan region of Rio de Janeiro, and reinforce the need for new epidemiological inquiries to clarify the participation of these in the dynamics of circulation of tick-borne pathogens.(AU)

Há uma crescente preocupação com a participação de hospedeiros e reservatórios silvestres na epidemiologia de diversos patógenos, principalmente no contexto das mudanças ambientais e da expansão do conceito "One Health". O objetivo deste estudo foi investigar a presença de hemoplasmas em gambás resgatados da região metropolitana do estado do Rio de Janeiro, Brasil. Amostras de sangue foram coletadas de 15 Didelphis aurita e submetidas à extração de DNA e PCR utilizando-se "primers" para os genes 16S rRNA e 23S rRNA. O exame físico e a análise hematológica também foram realizados. Três dos 15 gambás testaram positivo para Mycoplasma spp. hemotrópico por PCR. Os sinais clínicos eram inespecíficos e associados a lesões traumáticas. Anemia e leucocitose foram detectadas em animais positivos. A análise filogenética indicou que o hemoplasma detectado estava posicionado entre 'Ca. Mycoplasma haemodidelphis' detectado em D. virginiana da América do Norte e hemoplasmas recentemente detectados em D. aurita do estado de Minas Gerais, Brasil. Este estudo indica a existência de infecções por hemoplasmas em D. aurita, da região metropolitana do Rio de Janeiro, e reforça a necessidade de novos inquéritos epidemiológicos, para esclarecer a participação destes na dinâmica de circulação de patógenos transmitidos por carrapatos.(AU)

Cardiac hypertrophy associated with Corynebacterium spp. in Didelphis albiventris cubs

The purpose of this manuscript is to describe the clinical-pathological aspects of cardiac hypertrophy related to the presence of Corynebacterium spp. in three Didelphis albiventris cubs. In necropsy, macroscopically, in the heart, cardiomegaly, concentric hypertrophy of the ventricles and the ventricular septum were observed, with consequent reduction of the chamber. Microscopically, the primary lesions found in the heart were cardiomyocyte hypertrophy and necrosis, myocytolysis, and the presence of myriad basophilic bacteria. Liver fragments and endocardial swabs were sent for bacterial culture, in which pleomorphic Gram-positive rods grew, forming small and hemolytic colonies. Chemical tests demonstrated characteristics compatible with Corynebacterium spp. Thus, this report represents the first description of cardiac hypertrophy associated with Corynebacterium spp. in white-eared opossums cubs, representing an essential contribution to studying diseases in wild animals.(AU)

Lesão facial grave em um gambá-de-orelha-branca (Didelphis albiventris) - tratamento com ozonioterapia / Severe facial wound in a white-eared opossum (Didelphis albiventris) - treatment with ozone therapy

Background: Ozone therapy (OT) is a type of treatment that uses ozone gas, which has high oxidizing power. OT has been utilized for various diseases and to accelerate the wound healing process and reduce inflammation. Additionally, OT is a low-cost treatment recommended for patients that can be easily restrained. There are several methods of ozone administration, including topical use through bagging, cupping, or rectal insufflation, in addition to ozonated autohemotherapy. Based on the benefits of OT and the scarcity of reports describing its effects on white-eared opossums (Didelphis albiventris), we report on the use of ozone in the treatment of a severe facial wound in a white-eared opossum. Case: An adult female white-eared opossum weighing 1.6 kg was rescued from a fire. The patient was pregnant, dehydrated, hypothermic and had an extensive burn wound in the facial region. Due to the stress of captivity and its aggressive behavior, the animal needed to be chemically restrained with dexmedetomidine and butorphanol. Following sedation, the cephalic vein was catheterized and intravenous fluid therapy with ringer lactate was started to treat dehydration. Next, enrofloxacin, meloxicam, and tramadol hydrochloride were administered. Body rewarming was performed with thermal bags. The microbial load of the wound was reduced with chlorhexidine degerming solution followed by washing with saline solution. A thin layer of silver sulfadiazine cream was applied to the wound area. Over the following days the patient was treated with OT using the cupping technique combined with ozonated sunflower oil. After the first applications there was an improvement in the appearance of the wound, which initially had a purulent aspect. After 20 days of treatment the wound showed good granulation and there were no signs of infection, and at 45 days of treatment it was completely healed. The patient was then released with her baby. Discussion: Ozone therapy was included in the patient's treatment plan to enhance the tissue repair process in order to reintroduce the animal to the wild in the shortest possible time. The strategy was successful in improving healing and reducing tissue inflammation and infection. Ozone is known to cause the inactivation of microorganisms as a result of the oxidation of phospholipids and lipoproteins present in the cell membrane of bacteria and to interrupt viral replication. Ozone helps in the perfusion of injured tissues, contributing to the entry of humoral components, which control inflammation. Moreover, ozone contributes to increase supply of oxygen and nutrients to tissues and to stimulate increased production of fibrinogen and formation of granulation tissue. Ozone increases the enzymatic antioxidant response, while also playing a role in immunoregulation. Ozone can be used in several ways and via various methods with potential routes including intravenous, urethral, intramuscular, subcutaneous, or intra-articular, in addition to rectal and intramammary insufflation. Immunomodulatory, anti-inflammatory, analgesic, and antiseptic effects can be achieved with different doses and concentrations, which enables its application in different types of pathologies. Because of the aggressive behavior of the animal, in the current case we opted for the cupping technique combined with ozonated oil, both techniques which can be administered easily with only physical restraint of the patient. Cupping allows for the provision of a higher concentration of ozone gas locally whereas the ozonated oil maintains the effect of the gas for slightly longer, enhancing its effect and promoting tissue repair. The present report showed that OT is an effective alternative for the treatment of burn wounds in opossums, without causing adverse effects to the patient or her baby.

Molecular diversity of Sarcocystis spp. in opossums (Didelphis spp.) from Southeastern and Midwestern Brazil / Diversidade molecular de Sarcocystis spp. em gambás (Didelphis spp.) do Sudeste e Centro-oeste do Brasil

South American opossums (Didelphis spp.) are definitive hosts of Sarcocystis neurona, Sarcocystis speeri, Sarcocystis lindsayi and Sarcocystis falcatula. In Brazil, diverse studies have demonstrated a high frequency of Sarcocystis falcatula-like in sporocysts derived from opossums, and high genetic diversity has been observed in surface antigen-encoding genes (SAGs). In this study, genetic diversity of Sarcocystis spp. derived from Didelphis albiventris and Didelphis aurita from the cities of Campo Grande and São Paulo, was accessed by sequencing SAG2, SAG3, SAG4, the first internal transcribed spacer (ITS-1) and cytochrome c oxidase subunit I (cox1). Molecular identification was performed for 16 DNA samples obtained from sporocyst or culture-derived merozoites. The ITS-1, cox1, and SAG3 fragments were cloned, whereas SAG2 and SAG4 were sequenced directly from PCR products. Four alleles variants were found for SAG2, 13 for SAG3 and seven for SAG4, from which four, 13 and four, respectively, were novel. Twenty-seven allele variants were found for ITS-1, all phylogenetically related to S. falcatula-like previously described in Brazil. Sarcocystis sp. phylogenetically related to Sarcocystis rileyi was evidenced by cox1 in three opossums. Further studies are needed to clarify the role of Didelphis spp. as definitive hosts of Sarcocystis spp. other than that previous described.(AU)

Gambás sul-americanos (Didelphis spp.) são hospedeiros definitivos de Sarcocystis neurona, Sarcocystis speeri, Sarcocystis lindsayi e Sarcocystis falcatula. No Brasil, diversos estudos têm demonstrado alta frequência de Sarcocystis falcatula-like em esporocistos derivados de gambás, com grande diversidade nos genes que codificam antígenos de superfície (SAGs). Neste estudo, a diversidade genética de Sarcocystis spp., oriundos de Didelphis albiventris e Didelphis aurita, dos municípios de Campo Grande e São Paulo, foi acessada por meio do sequenciamento de SAG2, SAG3 e SAG4, da primeira região espaçadora interna transcrita (ITS-1) e citocromo c oxidase subunidade I (cox1). Identificação molecular foi realizada em 16 amostras de DNA, obtidas de esporocistos ou merozoítos derivados de cultivo. Os fragmentos de ITS-1, cox1 e SAG3 foram clonados, enquanto SAG2 e SAG4 foram sequenciados diretamente dos produtos de PCR. Quatro alelos foram observados em SAG2, 13 em SAG3 e sete em SAG4, sendo novos quatro, 13 e quatro, respectivamente. Em ITS-1, 27 alelos foram observados, todos filogeneticamente relacionados à S. falcatula-like, previamente detectados no Brasil. Sarcocystis sp. filogeneticamente relacionado à Sarcocystis rileyi foi evidenciado por cox1 em três gambás. Mais estudos são necessários para entender o papel de Didelphis spp. como hospedeiro definitivo de Sarcocystis spp. diferentes daqueles previamente descritos.(AU)

Molecular diagnostic for a screening investigation method of tick-borne pathogens in Didelphis albiventris road-killed in north of Paraná, Brazil

The synanthropization of wild animals puts public health at risk by promoting the circulation of zoonotic agents, found naturally in the wild, in the anthropic environment. The objective of this work was to carry out screening by molecular detection of pathogens of the Anaplasmatacea family in Didelphis albiventris, a specie characterized as having a synanthropic habit. Opossums that were dead (n = 25) after being road-killed were collected in the North of Paraná state, southern Brazil during the 2016 and 2018 years, through active search. A questionnaire was filled out with information about the animal and collected place. Biological samples of spleen and liver were collected. The genetic material extracted from the spleen and liver was submitted to molecular diagnosis through PCR for amplification of dsb of Ehrlichia and 16S genes for the other agents of the Anaplasmataceae family. One animal was positive for the genus Ehrlichia in semi-nested PCR for amplification of the 349 bp fragment of the dsb gene in extracted from the liver samples. In PCR for the 16S target no animal was positive. These are preliminary results that reinforce the circulation of Ehrlichia in opossums. To improve the knowledge of these agents in opossums more studies are necessary.(AU)

A sinantropização de animais silvestres coloca em risco a saúde pública por propiciar a circulação de agentes zoonóticos, encontrados naturalmente em ambiente silvestre, no ambiente antrópico. O trabalho teve como objetivo realizar a triagem por detecção molecular de patógenos da família Anaplasmataceae em Didelphis albiventris, espécie caracterizada como de hábito sinantrópico. Gambás mortos (n=25) por atropelamento durante os anos de 2016 e 2018 foram coletados na região norte do Paraná, sul do Brasil, por meio de busca ativa. Realizou-se o preenchimento de formulário com informações sobre a espécie do animal e o local do atropelamento. Foi realizada a necrópsia e coleta de amostras biológicas, de baço e fígado. O material genético extraído de baço e fígado foi submetido a diagnóstico molecular, por meio de PCR, para amplificação dos genes dsb de Ehrlichia sp. e 16S para os demais agentes da família Anaplasmataceae. Um animal foi positivo para o gênero Ehrlichia em semi-nested PCR para a amplificação do fragmento de 349 pb do gene dsb, extraído de fígado. Na PCR para detecção do gene 16S nenhum dos animais foi positivo. Esses resultados preliminares reforçam a circulação de Ehrlichias em gambás. Para melhorar o conhecimento desses agentes em gambás mais estudos são necessários.(AU)

Thoracic evaluation and cardiac measurement using radiography in opossums (Didelphis albiventris) / Avaliação radiográfica de toráx e mensuração cardíaca de gambás (Didelphis albiventris)

Cardiac measurement by chest X-ray is very important in the cardiac characterization of dogs and cats, however, the wild animal clinic has a different reality. Opossums (Didelphis albiventris) are part of the Brazilian fauna and are frequently referred for clinical and radiographic evaluation. The objective of this research was to evaluate the thorax of skunks and establish the average of VHS and VLAS for these animals. Chest radiographs were performed in lateral and ventrodorsal projections to measure the VHS and VLAS, thoracic depth and tracheal diameter. The mean VHS was 7.85 (± 0.3) for males and 8.12 (± 0.5) for females and the measured VLAS was 1.19 (± 0.2) for both sexes. The mean value for thoracic depth was 1.13 (± 0.03) and the ratio between the diameter of the chest inlet and the diameter of the trachea was 0.25 (± 0.02), with no significant differences between the sexes. This is the first study that addresses the thoracic evaluation by tracheal and cardiac measurements of skunks through radiographic examination.(AU)

A mensuração cardíaca por meio de radiografia de tórax é muito importante na caracterização cardíaca de cães e gatos, porém, a clínica de animais silvestres tem uma realidade diferente. Os gambás (Didelphis albiventris) fazem parte da fauna brasileira e são frequentemente encaminhados para avaliação clínica e para avaliação radiográfica. O objetivo desta pesquisa foi avaliar o tórax de gambás e estabelecer a média de VHS e VLAS para esses animais. Foram realizadas radiografias de tórax em projeções laterais e ventro dorsais, para mensuração do VHS e VLAS, a profundidade torácica e o diâmetro traqueal. O VHS médio foi de 7,85 (± 0,3) para os machos e 8,12 (± 0,5) para as fêmeas e o VLAS mensurado foi de 1,19 (± 0,2) para ambos os sexos. O valor médio da profundidade torácica foi 1,13 (± 0,03) e a relação entre o diâmetro da entrada do tórax e o diâmetro da traqueia foi de 0,25 (± 0,02), sem diferenças significativas entre os sexos. Este é o primeiro estudo que aborda a avaliação torácica a medida traqueal e cardíaca de gambás por meio de exame radiográfico.(AU)

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.

Helminth community structure of Didelphis marsupialis (Didelphimorphia, Didelphidae) in a transition area between the Brazilian Amazon and the Cerrado / Estrutura da comunidade de helmintos de Didelphis marsupialis (Didelphimorphia, Didelphidae) em uma área de transição entre a Amazônia brasileira e o Cerrado

Although the common opossum, Didelphis marsupialis (Didelphimorphia: Didelphidae) is a species widely distributed in South America, knowledge about their helminth parasites and helminth community structure is scarce. The aims of this study were to describe the species composition and analyze the structure of the helminth community of the common opossum in an area of the Amazonian Arc in northern Mato Grosso. The helminths were recovered, counted, and identified in 32 individuals. Overall, 10,198 specimens were categorized into 9 helminths taxa (seven nematodes, one cestode, and one acanthocephalan). The most abundant species were Aspidodera raillieti, Viannaia hamata, and Travassostrongylus orloffi. No statistically significant differences in helminth abundance and prevalence were observed between host sexes. However, young hosts had higher abundance and prevalence of Didelphonema longispiculata, whereas Oligacanthorhynchus microcephalus had higher abundance and prevalence in adult hosts. This was the first study to analyze the helminth fauna and helminth community structure of D. marsupialis in the Amazonian Arc. This is the first report of the presence of A. raillieti, D. longispiculata, T. orloffi, T. minuta, V. hamata, and O. microcephalus in the state of Mato Grosso, Brazil.(AU)

Embora o gambá comum, Didelphis marsupialis (Didelphimorphia: Didelphidae) seja uma espécie amplamente distribuída na América do Sul, o conhecimento sobre seus helmintos parasitos e a estrutura da comunidade de helmintos são escassos. Os objetivos deste estudo foram descrever a composição de espécies e analisar a estrutura da comunidade helmíntica do gambá comum em uma área do Arco Amazônico, no norte de Mato Grosso. Os helmintos foram recuperados, contados e identificados em 32 indivíduos. Ao todo, 10.198 espécimes foram categorizados em 9 táxons de helmintos (sete nematoides, um cestoide e um acantocéfalo). As espécies mais abundantes foram Aspidodera raillieti, Viannaia hamata e Travassostrongylus orloffi. Não foram observadas diferenças estatisticamente significativas na abundância e prevalência de helmintos entre os sexos dos hospedeiros. No entanto, hospedeiros jovens apresentaram maior abundância e prevalência de Didelphonema longispiculata, enquanto Oligacanthorhynchus microcephalus apresentou maior abundância e prevalência em hospedeiros adultos. Este foi o primeiro estudo a analisar a helmintofauna e a estrutura da comunidade helmíntica de D. marsupialis no Arco Amazônico. Este é o primeiro relato da presença de A. raillieti, D. longispiculata, T. orloffi, T. minuta, V. hamata e O. microcephalus no estado de Mato Grosso, Brasil.(AU)

Hemoparasites in Didelphis aurita from Seropédica, Rio de Janeiro state, Brazil / Hemoparasitos em Didelphis aurita capturados em Seropédica, estado do Rio de Janeiro, Brasi

The present study aimed to use polymerase chain reaction (PCR) to detect species of the order Piroplasmida, such as Anaplasma spp., Borrelia spp., and Ehrlichia spp., circulating in the blood of Didelphis aurita in a peridomiciliary environment. Blood samples collected from big-eared opossum (Didelphis aurita) were screened for hemoparasites using PCR. The extracted DNA was tested for tick-borne hemoparasites. We were unable to detect hemoparasites, such as Ehrlichia spp., Babesia spp., Anaplasma spp., and Borrelia spp. Theileria DNA was detected in only one sample screened using PCR for an approximately 650-base pair fragment of the 18S rRNA gene. Sequencing and BLAST analysis of a subset of the PCR amplicons revealed 97% (535/553 bp) identity with Theileria bicornis. The detection of Theileria sp. in D. aurita challenges us to pursue more in-depth studies of marsupial piroplasmosids and to evaluate the morphological aspects of the findings and their possible involvement in zoonoses.

O objetivo do presente estudo foi utilizar a reação em cadeia da polimerase (PCR) para detectar espécies da ordem Piroplasmida, como Anaplasma spp., Borrelia spp. e Ehrlichia spp., circulando no sangue de Didelphis aurita em ambiente peridomiciliar. Amostras de sangue coletadas de gambá da orelha preta (Didelphis aurita) foram triadas para hemoparasitas por meio da PCR. O DNA extraído foi testado para alguns hemoparasitas transmitidos por carrapatos. Não conseguimos detectar hemoparasitos, como Ehrlichia spp., Babesia spp., Anaplasma spp. e Borrelia spp. O DNA de Theileria foi detectado apenas em uma amostra. Um fragmento de ~650 pares de bases do gene 18S rRNA foi sequenciado. e a análise pelo BLAST (um subconjunto de amplicons de PCR) revelou uma identidade de 97% (535/553 pb) com Theileria bicornis. A detecção de Theileria sp. in D. aurita nos desafia a aprofundar estudos sobre piroplasmosídeos marsupiais e avaliar os aspectos morfológicos dos achados e seu possível envolvimento com zoonoses.

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.

Electrocardiographic evaluation of baby opossum (Didelphis albiventris) / Avaliação eletrocardiográfica de gambás bebê (Didelphis albiventris)

O eletrocardiograma (ECG) é um importante método diagnóstico, essencial para determinar o ritmo e o funcionamento elétrico do coração. Os gambás (D. albiventris) frequentemente são atendidos nos Centros de Triagem de Animais Silvestres (CETAS), contudo pouco se sabe sobre o sistema cardiovascular dessa espécie. Dessa forma, o objetivo foi estabelecer valores de referência dos parâmetros eletrocardiográficos dessa espécie. Foram utilizados 32 filhotes, considerados hígidos nos exames clínicos geral e específico. Avaliaram-se a frequência cardíaca, o ritmo, a morfologia, a amplitude e a duração das ondas P, o complexo QRS e T, a duração dos intervalos PR e QT e o segmento ST. A frequência cardíaca média foi de 214 batimentos por minuto (bpm), o ritmo sinusal mostrou-se normal e houve arritmia sinusal em 25% dos pacientes. O eixo médio foi de 40º(QRS)e 47º (P). A onda P apresentou morfologia positiva (81,3%) e negativa (18,7%), amplitude de 0,03mV (±0,01) e duração de 0,04s (±0,04). O complexo QRS apresentou morfologia QRS ou RS (31%) e duração de 0,05s (±0,06). A amplitude de Q foi de -0,04mV (±0,05), a de R foi 0,23mV (±0,16) e S -0,07mV (±0,08). Já a onda T apresentou morfologia variada, amplitude de 0,04mV (±0,04) e duração de 0,04s (±0,09). O intervalo PR obteve duração de 0,05s (±0,08), QT com intervalo entre 0,08 e 0,21 e o segmento ST médio de 0,05s (±0,02). Dessa forma, observou-se que foi possível realizar o ECG em gambás filhotes e que o sexo e o peso não influenciaram no traçado obtido. Os resultados diferiram de cães, gatos e de outras espécies de gambás.

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.(AU)

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.(AU)

Sarcocystis neurona and related Sarcocystis spp. shed by opossums (Didelphis spp.) in South America / Sarcocystis neurona e Sarcocystis spp. relacionados, excretados por gambás (Didelphis spp.) na América do Sul

Protozoan parasites of the genus Sarcocystis are obligatory heteroxenous cyst-forming coccidia that infect a wide variety of animals and encompass approximately 200 described species. At least four Sarcocystis spp. (S. falcatula, S. neurona, S. lindsayi and S. speeri) use opossums (Didelphis spp.) as definitive hosts, and two of them, S. neurona and S. falcatula, are known to cause disease in horses and birds, respectively. Opossums are restricted to the Americas, but their distribution in the Americas is heterogeneous. Five Didelphis spp. are distributed in South America (D. aurita, D. albiventris, D. marsupialis, D. imperfecta and D. pernigra) whereas just one opossum species (D. virginiana) is found in North America. Studies conducted in the last decades show that Sarcocystis spp., derived from South American Didelphis spp., have biological and genetic differences in relation to Sarcocystis spp. shed by the North American opossum D. virginiana. The aim of this review was to address the peculiar scenario of Sarcocystis species shed by South American opossums, with a special focus on diagnosis, epidemiology, and animal infections, as well as the genetic characteristics of these parasites.(AU)

Parasitos protozoários do gênero Sarcocystis são coccídios heteroxenos formadores de cistos, que infectam variadas espécies animais e compreendem cerca de 200 espécies descritas. Pelo menos quatro Sarcocystis spp. (S. falcatula, S. neurona, S. lindsayi e S. speeri) utilizam gambás (Didelphis spp.) como hospedeiros definitivos; e duas delas, S. neurona and S. falcatula são conhecidas por causarem doença em equinos e aves, respectivamente. Gambás didelfídeos são restritos ao continente americano, contudo são distribuídos de forma heterogênea nas Américas. Cinco Didelphis spp. são distribuídos na América do Sul (D. aurita, D. albiventris, D. marsupialis, D. imperfecta e D. pernigra), enquanto somente uma espécie (D. virginiana) é encontrada na América do Norte. Trabalhos conduzidos, nas últimas décadas, mostram que Sarcocystis spp. derivados de Didelphis spp. sul-americanos possuem diferenças biológicas e genéticas, quando comparados a Sarcocystis spp. excretados pelo gambá norte-americano D. virginiana. O objetivo desta revisão é discutir a situação peculiar das espécies de Sarcocystis na América do Sul com um foco especial em diagnóstico, epidemiologia e infecções animais, assim como nas características genéticas desses parasitos.(AU)

Mamíferos em remanescentes florestais de mata atlântica, Barbacena, Minas Gerais / Mammals in atlantic forest remnants of Barbacena, Minas Gerais

A perda e fragmentação de habitats são as principais causas do declínio da biodiversidade, uma vez que as alterações das condições ambientais e ecológicas afetam diretamente a riqueza, a abundância e a distribuição das espécies. O presente trabalho teve como objetivo identificar mamíferos de médio e grande porte em três fragmentos remanescentes de Mata Atlântica, situados no Campus Barbacena do Instituto Federal do Sudeste de Minas Gerais (IFSudesteMG). As espécies foram registradas por meio de parcelas de areia e busca ativa de vestígios diretos e indiretos. Foram identificadas 16 espécies distribuídas em seis ordens e 14 famílias, dentre elas, algumas vulneráveis e ameaçadas como Lycalopex vetulus (raposa do campo), Lontra longicaudis (lontra) e Callicebus nigrifrons (sauá). Canis familiaris (cão doméstico) e Didelphis sp. (gambá) foram mais frequentes nas parcelas de areia. O maior fragmento apresentou maior riqueza, com 15 espécies, 93,75% do total registrado no Campus. O número de espécies compartilhadas nas três áreas amostradas foi 31,2%. Os índices de similaridade registrados entre os fragmentos indicam que há necessidade de maior conectividade entre as áreas através da criação de corredores ecológicos, permitindo assim o deslocamento das espécies e, consequentemente, o maior fluxo gênico, favorecendo a manutenção de importantes serviços ecossistêmicos para a região. Apesar de pequena, a área de estudo desempenha importante papel na conservação dos mamíferos da região, preservando espécies de Cerrado e Mata Atlântica. Com o intuito de reduzir os impactos na fauna local, torna-se necessário a implantação de ações de manejo e conservação.

Habitat loss and fragmentation are the main causes of biodiversity decline, since changes in environmental and ecological conditions directly affect species richness, abundance, and distribution. This study aimed to identify medium- and large-sized mammals occurring in three Atlantic Forest remnants located in the Barbacena campus of the Federal Institute of Southeast Minas Gerais (IFSudesteMG). The species were recorded in track plots and through active search for direct and indirect signs. 16 species distributed in six orders and 14 families were identified, including some vulnerable and threatened species such as Lycalopex vetulus (meadow fox), Lontra longicaudis (otter), and Callicebus nigrifrons (black-fronted titi monkey). Canis familiaris (domestic dog) and Didelphis sp. (opossum) were the species most frequently observed in the track plots. The largest fragment had the greatest species richness, with 15 species, or 93.75% of the total. The number of species shared among the three sampled areas was 31.2%. The similarities identified between the fragments indicate the need for higher connectivity among them, through the creation of ecological corridors to allow the movement and, consequently, gene flux among the populations, favoring the maintenance of important ecosystem services in the region. Although small, the study area plays a major role in mammal conservation in the region, preserving species from the Cerrado and the Atlantic Forest domains. To reduce potential impacts on the local fauna, management and conservation efforts should be put forward.

Small mammals from the lasting fragments of Araucaria Forest in southern Brazil: a study about richness and diversity

Rodents and marsupials together, form the most rich group of mammals in Brazil, they are distributed in all biomes, including Atlantic Forest, and can be used to analyze environmental quality. The Araucaria Forest is a threatened formation of the Atlantic forest with few areas remaining intact and a huge lack of knowledge about the potential of its secondary forest fragments to act as habitat refuges to native fauna. This study evaluated the species richness and diversity of non-volant small mammals among five vegetation types within a protected area in an Araucaria Forest area from southern Brazil. The sampling was made in six bimonthly campaigns during a year, by pitfall and live traps. This effort resulted in 875 captures of 625 individuals from 16 species. The most common species in Pine Plantation was Oligoryzomys nigripes, in Natural Regeneration forest was Thaptomys nigrita and Akodon montensis was most often captured in the other vegetation. Although differences in species composition were found among the different vegetation types, there was no significant difference in species richness. Nevertheless, true diversity values were significantly lower in the Pine Plantation. The results demonstrated that the substitution of native for exotic vegetation generated a negative impact on the group diversity. More studies, in other Araucaria Forest areas are necessary to allow better understanding of the processes driving this loss of diversity in this biome threatened by antropic action.(AU)

Small mammals from the lasting fragments of Araucaria Forest in southern Brazil: a study about richness and diversity

Rodents and marsupials together, form the most rich group of mammals in Brazil, they are distributed in all biomes, including Atlantic Forest, and can be used to analyze environmental quality. The Araucaria Forest is a threatened formation of the Atlantic forest with few areas remaining intact and a huge lack of knowledge about the potential of its secondary forest fragments to act as habitat refuges to native fauna. This study evaluated the species richness and diversity of non-volant small mammals among five vegetation types within a protected area in an Araucaria Forest area from southern Brazil. The sampling was made in six bimonthly campaigns during a year, by pitfall and live traps. This effort resulted in 875 captures of 625 individuals from 16 species. The most common species in Pine Plantation was Oligoryzomys nigripes, in Natural Regeneration forest was Thaptomys nigrita and Akodon montensis was most often captured in the other vegetation. Although differences in species composition were found among the different vegetation types, there was no significant difference in species richness. Nevertheless, true diversity values were significantly lower in the Pine Plantation. The results demonstrated that the substitution of native for exotic vegetation generated a negative impact on the group diversity. More studies, in other Araucaria Forest areas are necessary to allow better understanding of the processes driving this loss of diversity in this biome threatened by antropic action.

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

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

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

Sarcocystis neurona and related Sarcocystis spp. shed by opossums (Didelphis spp.) in South America / Sarcocystis neurona e Sarcocystis spp. relacionados, excretados por gambás (Didelphis spp.) na América do Sul

Abstract Protozoan parasites of the genus Sarcocystis are obligatory heteroxenous cyst-forming coccidia that infect a wide variety of animals and encompass approximately 200 described species. At least four Sarcocystis spp. (S. falcatula, S. neurona, S. lindsayi and S. speeri) use opossums (Didelphis spp.) as definitive hosts, and two of them, S. neurona and S. falcatula, are known to cause disease in horses and birds, respectively. Opossums are restricted to the Americas, but their distribution in the Americas is heterogeneous. Five Didelphis spp. are distributed in South America (D. aurita, D. albiventris, D. marsupialis, D. imperfecta and D. pernigra) whereas just one opossum species (D. virginiana) is found in North America. Studies conducted in the last decades show that Sarcocystis spp., derived from South American Didelphis spp., have biological and genetic differences in relation to Sarcocystis spp. shed by the North American opossum D. virginiana. The aim of this review was to address the peculiar scenario of Sarcocystis species shed by South American opossums, with a special focus on diagnosis, epidemiology, and animal infections, as well as the genetic characteristics of these parasites.

Resumo Parasitos protozoários do gênero Sarcocystis são coccídios heteroxenos formadores de cistos, que infectam variadas espécies animais e compreendem cerca de 200 espécies descritas. Pelo menos quatro Sarcocystis spp. (S. falcatula, S. neurona, S. lindsayi e S. speeri) utilizam gambás (Didelphis spp.) como hospedeiros definitivos; e duas delas, S. neurona and S. falcatula são conhecidas por causarem doença em equinos e aves, respectivamente. Gambás didelfídeos são restritos ao continente americano, contudo são distribuídos de forma heterogênea nas Américas. Cinco Didelphis spp. são distribuídos na América do Sul (D. aurita, D. albiventris, D. marsupialis, D. imperfecta e D. pernigra), enquanto somente uma espécie (D. virginiana) é encontrada na América do Norte. Trabalhos conduzidos, nas últimas décadas, mostram que Sarcocystis spp. derivados de Didelphis spp. sul-americanos possuem diferenças biológicas e genéticas, quando comparados a Sarcocystis spp. excretados pelo gambá norte-americano D. virginiana. O objetivo desta revisão é discutir a situação peculiar das espécies de Sarcocystis na América do Sul com um foco especial em diagnóstico, epidemiologia e infecções animais, assim como nas características genéticas desses parasitos.

Toxoplasmose encefálica em um gambá-de-orelha-branca (Didelphis albiventris) / Encephalic toxoplasm in a white-eared possum (Didelphis albiventris)

Background: Toxoplasmosis is caused by Toxoplasma gondii, an obligate intracellular protozoan that belongs to the Apicomplexa phylum, coccidian subclass, and affects all warm-blooded animals. The role of opossums in the epidemiology of toxoplasmosis in Brazil is not fully understood, and there are very few descriptions of toxoplasmosis lesions in these animals. This report describes the anatomopathological, molecular and immunohistochemical findings of a case of encephalic toxoplasmosis in free-living white-eared possum (Didelphis albiventris). Case: A young male opossum (D. albiventris), was treated at the Veterinary Hospital of Wild Animals of the University of Brasília, Federal District. The animal was apathetic, uncoordinated, reluctant to move, and had an exposed proximal fracture in the left radius and ulna with laceration of muscles and adjacent tendinous structures. Amputation on the left thoracic limb was performed followed by analgesia and antibiotic therapy. The environment is frequented by other wild animals, and stray cats have access to the patio of the building. Twenty-five days after arriving at the hospital, the animal was found dead in its cage. After death, a necropsy was performed. Organ fragments from the abdominal cavity, thoracic and central nervous system were collected, processed routinely for histology and stained with hematoxylin and eosin. Macroscopic lesions in the central nervous system were not observed. On microscopy, the brain showed moderate random glial nodules throughout the neuropil associated with the presence of spherical to elongated parasitic cysts of about 20 µm, with a thin wall and with its interior full of bradyzoites, consistent with Toxoplasma gondii. There was also moderate fibrinoid necrosis and moderate multifocal lymphoplasmacytic infiltrate surrounding the blood vessels (perivascular cuffs) To investigate the etiology of the brain injury, brain sections were subjected to immunohistochemistry (IHC) and real-time polymerase chain reaction (qPCR) technique for detection of T. gondii and Neospora caninum. Immunostaining for T. gondii in the cyst wall and in bradyzoites and negative immunostaining for N. caninum. qPCR was positive for T. gondii and negative for N. caninum. Discussion: Diagnosis of encephalic toxoplasmosis in a Didelphis albiventris was possible based on histopathological, immunohistochemical and molecular findings. The morphological classification of the brain lesion was important for the diagnosis. Brain toxoplasmosis in opossums usually results in focal areas of malacia on macroscopy and focally extensive necrosis on microscopy, neutrophil infiltrate, calcified necrotic material, and perivascular cuffs of lymphocytes and plasma cells. In the present case, similar histopathological lesions were noted, but no significant macroscopic changes were observed. The etiology here was defined by immunohistochemistry and qPCR, techniques proven to be useful and with good specificity for diagnosing toxoplasmosis in mammals. It is believed that the positive immunohistochemical and molecular result for Toxoplasma gondii together with the negative result for Neospora caninum were conclusive for the diagnosis. Thus, we demonstrate here a post mortem diagnosis of toxoplasmosis in a free-living synanthropic opossum and the use of anatomopathology, immunohistochemistry and real-time polymerase chain reaction as a diagnostic option for this disease in opossums.

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.