Neorickettsia sp. in coatis (Nasua nasua) in Brazil / Neorickettsia sp. em quatis (Nasua nasua) no Brasil

The genus Neorickettsia comprises trematode-associated bacteria that can cause diseases in animals and humans. Despite detection of Neorickettsia antigens in the intestine of coatis kept in captivity in southern Brazil through immunohistochemistry, the molecular identity of the bacteria in South American procyonids remains elusive. The aim of the present study was to investigate the occurrence of Neorickettsia sp. in blood samples from coatis in central-western Brazil. Between March 2018 and January 2019, animals were captured and recaptured in two areas of the Cerrado (Parque Estadual do Prosa, PEP; and Vila da Base Aérea, VBA) located in the city of Campo Grande, state of Mato Grosso do Sul, central-western Brazil. All captures were performed according to convenience. DNA from 97 blood samples was subjected to nested PCR (nPCR) targeting a fragment of the 16S rRNA gene of Neorickettsia sp. Six samples (3.6%; five from VBA and one from PEP) from different coatis were positive in nPCR based on the 16S rRNA. The sequences obtained (~500 bp) showed ˃ 99% similarity to N. risticii. Phylogenetic analysis clustered the sequences detected in the present study in a clade with N. risticii. This is the first molecular detection of Neorickettsia sp. in coatis in Brazil.(AU)

O gênero Neorickettsia compreende bactérias associadas a trematódeos que podem causar doenças em animais e humanos. Apesar da detecção de antígenos de Neorickettsia por imuno-histoquímica no intestino de quatis mantidos em cativeiro no sul do Brasil, a identidade molecular da bactéria em procionídeos da América do Sul permanece indefinida. O presente trabalho teve como objetivo investigar a ocorrência de Neorickettsia sp. em amostras de sangue de quatis do Centro-Oeste do Brasil. Entre março de 2018 e janeiro de 2019, os animais foram capturados em duas áreas de Cerrado (Parque Estadual do Prosa PEP e Vila da Base Aérea VBA) localizadas na cidade de Campo Grande, Mato Grosso do Sul, Centro-Oeste do Brasil. Todas as capturas e recapturas foram realizadas por conveniência. O DNA de 97 amostras de sangue foi submetido a "nested" (nPCR), baseada em um fragmento do gene 16S rRNA de Neorickettsia sp. Seis (3,6% - 5 de VBA e 1 de PEP) amostras de quatis diferentes foram positivas na nPCR, baseada no rRNA 16S. As sequências obtidas (~500 pb) apresentaram ˃99% de identidade com N. risticii. A inferência filogenética agrupou as sequencias detectadas no presente estudo em um clado com N. risticii. Esta é a primeira detecção molecular de Neorickettsia sp. em quatis do Brasil.(AU)

Genetic diversity of Hepatozoon spp. in rodents from Chile / Diversidade genética de Hepatozoon spp. em roedores do Chile

Abstract This study aimed to investigate the genetic diversity of Hepatozoon spp. in rodents from Valdivia, Chile. A total of 74 rodents (synanthropic n=38; wild n=36) were trapped in Valdivia. We performed conventional PCR assays for Apicomplexa organisms targeting two overlapping 18S rDNA gene fragments (600 bp and 900 bp) followed by sequencing of selected amplicons. Hepatozoon spp. occurrence was 82.43% (61/74). Twelve sequences obtained from the 600 bp and ten from the 900 bp 18S rDNA fragments were identified as Hepatozoon sp. Six sequences obtained from 18S rDNA-based overlapping PCR protocols were used for concatenated (1,400 bp) phylogenetic, haplotype and distance analyses. Hepatozoon spp. 18S rDNA concatenated sequences from the present study were detected in Oligoryzomys longicaudatus, Rattus norvegicus, Mus musculus, and Abrothrix longipilis grouped with Hepatozoon species earlier described in rodents and reptiles from Chile and Brazil. Nucleotide polymorphism of the six 18S rDNA sequences (1,400 bp) from this study, and other Chilean sequences from rodents and rodent's ticks, showed high diversity with a total of nine Chilean haplotypes. Three haplotypes from Valdivia were identified for the first time in this study, suggesting the circulation of novel haplotypes in rodents from southern Chile.

Resumo Este estudo teve como objetivo investigar a diversidade genética de Hepatozoon spp. em roedores de Valdivia, Chile. Um total de 74 roedores (sinantrópicos n=38; selvagens n=36) foram capturados. PCR convencional foi realizada para organismos Apicomplexa, visando dois fragmentos sobrepostos do gene 18S rDNA (600 bp e 900 bp), seguida pelo sequenciamento de amplicons selecionados. A ocorrência de Hepatozoon spp. foi de 82,43% (61/74). Doze sequências obtidas dos fragmentos de 18S rDNA de 600 pb e dez dos fragmentos de 18S rDNA de 900 pb foram identificadas como Hepatozoon sp. Seis sequências obtidas, a partir de protocolos de PCR sobrepostos, foram usadas para análises filogenéticas (1.400 bp), de haplótipos e de distância. Sequências concatenadas 18S rDNA do presente estudo foram detectadas em Oligoryzomys longicaudatus, Rattus norvegicus, Mus musculus e Abrothrix longipilis e agrupadas com Hepatozoon descrito em roedores e répteis do Chile e do Brasil. A análise de polimorfismos das seis sequências deste estudo, junto com outras sequências chilenas de roedores e carrapatos de roedores, mostrou alta diversidade com um total de nove haplótipos no Chile. Três haplótipos detectados em Valdivia foram identificados pela primeira vez neste estudo, sugerindo que novos haplótipos circulam em roedores do sul do Chile.

Hepatozoon sp. gamonts as an accidental finding in synovial liquid from an injured maned wolf (Chrysocyon brachyurus) in southeastern Brazil / Gamontes de Hepatozoon sp. como achado acidental em líquido sinovial de lobo guará (Chrysocyon brachyurus) no sudeste do Brasil

A free-living, adult male maned wolf (Chrysocyon brachyurus) was referred to the Governador Laudo Natel FCAV/Unesp veterinary hospital after being found with skin lesions and a fracture on the right pelvic limb, which had to be amputated due to compromised integrity. Around 20 days later, bilateral accentuated swollen on humerus-radius-ulna articulation was observed. The synovial liquid was drained and sent to the laboratory for synovial cytology with Rosenfeld staining that revealed predominantly degenerated neutrophils with karyolytic chromatin associated with intracellular inclusions suggestive of Hepatozoon sp. gametocytes. Blood and synovial liquid samples were submitted to molecular analysis, aiming to amplify the Hepatozoon spp. 18S rRNA gene fragment. Despite the positioning of the found Hepatozoon sequence together with Hepatozoon canis previously detected in domestic carnivores, the BLAST analysis showed only 98% identity with H. canis. To the best of the authors knowledge, this is the first time a Hepatozoon was detected in the synovial liquid by clinical pathology and molecular analyses.(AU)

Um lobo guará (Chrysocyon brachyurus) adulto, macho, de vida livre foi encaminhado para atendimento no hospital veterinário Governador Laudo Natel FCAV/Unesp após ser encontrado com lesões de pele e fratura em membro pélvico direito, sendo amputado devido a comprometimento da integridade do membro. Aproximadamente 20 dias após a chegada ao hospital, foi notado acentuado aumento de volume bilateral em região de articulação úmero-rádio-ulnar. O líquido sinovial foi drenado e enviado para análise citológica com coloração de Rosenfeld, revelando a presença de neutrófilos degenerados com cromatina cariolítica associados a inclusões intracelulares sugestivas de gametócitos de Hepatozoon sp. Amostras de sangue e líquido sinovial foram submetidas a análises moleculares visando amplificar um fragmento do gene 18S rRNA de Hepatozoon spp. Apesar da sequência de Hepatozoon detectada se posicionar filogeneticamente no mesmo clado que H. canis previamente detectado em carnívoros domésticos, o resultado da análise do BLAST mostrou somente 98% de identidade com H. canis. De acordo com o conhecimento dos autores, esta é a primeira vez que Hepatozoon foi detectado no líquido sinovial por meio de patologia clínica e análises moleculares.(AU)

Association of rush pin and intramedular pin techniques for the stabilization of salter harris type I fracture in rabbit (Oryctolagus cuniculus)

Background: In domestic rabbits, fractures are usually the result of household accidents. Fractures of the distal femoral physis are frequently observed in animals with immature skeletons and may cause future orthopedic problems. With this type of fracture, early reduction and stabilization are necessary to prevent additional damage to the physis and to preserve the growth potential of the bone. This report aims to describe the clinical and radiographic findings, as well as the surgical method used that combined Rush pins with an intramedullary pin, associated with a Salter Harris type I fracture of the right distal femur in a rabbit.Case: A 4-month male domestic rabbit (Oryctolagus cuniculus) weighing 1.2 kg, was referred to the Veterinary Hospital with a right pelvic limb lameness after an episode of trauma. Orthopedic examination revealed swelling, instability, crepitation, and increased pain sensitivity in the distal aspect of the right femur. Following physical and radiographic examination, a Salter Harris type I fracture of the right distal femur was diagnosed. The patient was premedicated with a combination of xylazine (2 mg/kg IM) and ketamine (10 mg/kg IM), which allowed placement of an IV catheter for the administration of fluids and intubation using an endotracheal tube with an internal diameter of 2 mm. Anesthesia was maintained with isoflurane, and an epidural was performed with a combination of lidocaine (2 mg/kg) and bupivacaine (0.75 mg/kg). Osteosynthesis was performed with two Rush pins and an intramedullary pin. Radiographic examination 110 and 330 days after the surgical procedure showed good alignment of the bone and adequate healing of the fracture. The combination of techniques used in this case report proved to be effective, resulting in functional recovery of the limb and rapid bone healing.[...](AU)

Association of rush pin and intramedular pin techniques for the stabilization of salter harris type I fracture in rabbit (Oryctolagus cuniculus)

Background: In domestic rabbits, fractures are usually the result of household accidents. Fractures of the distal femoral physis are frequently observed in animals with immature skeletons and may cause future orthopedic problems. With this type of fracture, early reduction and stabilization are necessary to prevent additional damage to the physis and to preserve the growth potential of the bone. This report aims to describe the clinical and radiographic findings, as well as the surgical method used that combined Rush pins with an intramedullary pin, associated with a Salter Harris type I fracture of the right distal femur in a rabbit.Case: A 4-month male domestic rabbit (Oryctolagus cuniculus) weighing 1.2 kg, was referred to the Veterinary Hospital with a right pelvic limb lameness after an episode of trauma. Orthopedic examination revealed swelling, instability, crepitation, and increased pain sensitivity in the distal aspect of the right femur. Following physical and radiographic examination, a Salter Harris type I fracture of the right distal femur was diagnosed. The patient was premedicated with a combination of xylazine (2 mg/kg IM) and ketamine (10 mg/kg IM), which allowed placement of an IV catheter for the administration of fluids and intubation using an endotracheal tube with an internal diameter of 2 mm. Anesthesia was maintained with isoflurane, and an epidural was performed with a combination of lidocaine (2 mg/kg) and bupivacaine (0.75 mg/kg). Osteosynthesis was performed with two Rush pins and an intramedullary pin. Radiographic examination 110 and 330 days after the surgical procedure showed good alignment of the bone and adequate healing of the fracture. The combination of techniques used in this case report proved to be effective, resulting in functional recovery of the limb and rapid bone healing.[...]

Salivary gland adenocarcinoma in an orange-spined hairy dwarf porcupine (Sphiggurus villosus)

The goal of this case report is to describe a salivary gland adenocarcinoma in a free ranging Sphiggurus villosus. A orange-spined hairy dwarf porcupine (S. villosus) was referred to the Wildlife Medicine Service at the Veterinarian Hospital of the Universidade Estadual Paulista (Jaboticabal, São Paulo, Brazil), with a mass in the lateral region of the face. Radiographs highlighted swelling in the soft tissues, measuring 7 cm x 9 cm, without bone involvement. The physical examination and radiographs were consistent with a neoplastic process. The animal was submitted to surgical excision of the mass. Microscopically, the tumor was composed of a highly cellular proliferation of epithelial cells arranged predominantly in tubular pattern, associated with moderated fibrous stroma, poorly demarcated, non-encapsulated and with infiltrative growth. These findings were consistent with salivary gland adenocarcinoma. To the authors" knowledge, this is the first report of a salivary gland adenocarcinoma in a wild life orange-spined hairy dwarf porcupine.

Salivary gland adenocarcinoma in an orange-spined hairy dwarf porcupine (Sphiggurus villosus)

The goal of this case report is to describe a salivary gland adenocarcinoma in a free ranging Sphiggurus villosus. A orange-spined hairy dwarf porcupine (S. villosus) was referred to the Wildlife Medicine Service at the Veterinarian Hospital of the Universidade Estadual Paulista (Jaboticabal, São Paulo, Brazil), with a mass in the lateral region of the face. Radiographs highlighted swelling in the soft tissues, measuring 7 cm x 9 cm, without bone involvement. The physical examination and radiographs were consistent with a neoplastic process. The animal was submitted to surgical excision of the mass. Microscopically, the tumor was composed of a highly cellular proliferation of epithelial cells arranged predominantly in tubular pattern, associated with moderated fibrous stroma, poorly demarcated, non-encapsulated and with infiltrative growth. These findings were consistent with salivary gland adenocarcinoma. To the authors" knowledge, this is the first report of a salivary gland adenocarcinoma in a wild life orange-spined hairy dwarf porcupine.(AU)

Hepatozoon caimani in Caiman crocodilus yacare (Crocodylia, Alligatoridae) from North Pantanal, Brazil / Hepatozoon caimani em Caiman crocodilus yacare (Crocodylia, Alligatoridae) do Norte do Pantanal, Brasil

Hepatozoon species are the most common intracellular hemoparasite found in reptiles. Hepatozoon caimani, whose vectors are Culex mosquitoes, has been detected in a high prevalence among caimans in Brazil by blood smears examinations. The present work aimed to detect and characterize the Hepatozoon spp. found in 33 caimans (24 free-ranging and 9 captive; 28 males and 5 females) (Caiman crocodilus yacare) sampled at Poconé, North Pantanal, state of Mato Grosso, Brazil, using blood smears examinations and molecular techniques. Hepatozoon spp.-gametocytes were found in 70.8% (17/24) and 88.8% (8/9) of blood smears from free-ranging and captive caimans, respectively. Hepatozoon spp. 18S rRNA DNA was found in 79.2% (19/24) and 88.8% (8/9) of free-ranging and captive caimans, respectively. Comparative analysis of parasitized and non-parasitized erythrocytes showed that all analyzed features were significantly different (P 0.05) for both linear and area dimensions. Phylogenetic analysis based on 18S rRNA sequences grouped the Hepatozoon spp. sequences detected in the present study together with H. caimani, recently detected in caimans in southern Pantanal.(AU)

Espécies do gênero Hepatozoon são os hemoparasitas intracelulares mais comumente encontrados em répteis. Hepatozoon caimani, cujos vetores são mosquitos do gênero Culex sp., têm sido detectados em uma alta prevalência entre jacarés no Brasil, por meio da análise de esfregaços sanguíneos. O presente estudo objetivou detectar e caracterizar parasitas do gênero Hepatozoon spp. em 33 jacarés (24 de vida-livre e 9 de cativeiro; 28 machos e 5 fêmeas) (Caiman crocodilus yacare) amostrados em Poconé, região norte do Pantanal, estado do Mato Grosso, Brasil, por meio da análise de esfregaços sanguíneos e técnicas moleculares. Gametócitos de Hepatozoon spp. foram encontrados em 70,8% (17/24) e em 88,8% (8/9) dos esfregaços sanguíneos de jacarés de via-livre e cativeiro, respectivamente. 18S rRNA DNA de Hepatozoon spp. foi detectado em 79,2% (19/24) e 88,8% (8/9) das amostras de sangue de jacarés de vida-livre e cativeiro, respectivamente. A análise comparativa de eritrócitos parasitados e não parasitados mostrou diferença significativa (P 0,05) em todas as variáveis lineares e de área analisadas. A análise filogenética baseada em sequências de DNA do 18S rRNA agrupou as sequências de Hepatozoon spp. detectadas no presente estudo juntamente com aquelas de H. caimani, recentemente detectadas em jacarés do Pantanal do Mato Grosso do Sul.(AU)