Transferabilidade de iniciadores de regiões microssatélites produzidos em veado catingueiro (Mazama gouazoubira) para uso em estudos genéticos com cervos-do-pantanal (Blastocerus dichotomus)

O cervo-do-pantanal (Blastocerus dichotomus), único representante do gênero Blastocerus é a maior espécie de cervídeo neotropical. A espécie é classificada pela lista vermelha da IUCN como vulnerável, especialmente pela perda de seu habitat, a várzea, para agropecuária e hidrelétricas. Com isso há grande frequência de isolamento populacional e consequentemente problemas com deriva e endogamia nas pequenas populações remanescentes. Os estudos genéticos podem contribuir de maneira decisiva no entendimento do efeito do isolamento populacional na sustentabilidade genética das populações. O presente estudo teve como objetivo testar iniciadores heterólogos desenvolvidos para a espécie Mazama gouazoubira em amostras de cervos-do-pantanal de uma população reintroduzida e com baixo nível de diversidade genética. O DNA de 8 animais reintroduzidos foi amplificado por meio de iniciadores e enviados para genotipagem. Sete dos 16 iniciadores heterólogos selecionados, amplificaram a região microssatélite esperada, sendo somente dois deles polimórficos, confirmando a baixa diversidade genética desta população. A partir dos resultados obtidos, o estudo permitiu identificar iniciadores heterólogos produzidos para Mazama gouazoubira capazes de amplificar a mesma região microssatélite para os cervídeos da espécie Blastocerus dichotomus

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.Discussion: Preoperative planning for orthopedic surgery in rabbits is different from that of dogs and cats, due to the particularities of the species. Familiarity with the regional anatomy, patient preparation, and appropriate instrumentation are necessary when contemplating osteosynthesis in a rabbit. Salter Harris fractures affect young animals, where the physis is considered an area of fragility in the bone. Surgical planning should take into account the function of the growth plates.It is recommended that implants passing through the physis do so perpendicularly, since angulation greater than 45° may predispose the bone to premature closure of the growth plate. Another important consideration involves the choice of implants, since the use of trocar-tipped pins facilitates their precise placement in the bone. The use of threaded pins should be avoided due to their weakness at the thread-shaft interface, and the risk of impaired longitudinal bone growth and thedifficulty of removal if necessary. Steinman pins and Kirschner wires can be used to stabilize a variety of different fractures. In Salter-Harris type I and II fractures, the use of pins neutralizes bending forces but not rotational or compressive forces. In contrast, Rush pinning and cross-pinning techniques are effective in neutralizing the forces acting on the physis, and are frequently used for the fixation of fractures in this region. Complications, often associated with poor reduction and alignment, can result in varus or valgus deviation and are associated with a high risk of implant failure or migration, malunion, and patellar luxation. It was concluded that the combination of Rush pins and an intramedullary pin resulted in adequate stabilization of the Salter Harris type I fracture of the distal femur in this rabbit.Keywords: Orthopedic implants, osteosynthesis, trauma, bone repairing, lagomorph

Theileria terrestris nov. sp. : a novel Theileria in lowland tapirs (Tapirus terrestris) from two different biomes in Brazil

The low-land tapir (Tapirus terrestris) is the largest wild terrestrial mammal found in Brazil. Although T. terrestris has been already reported as a host of hemoparasites, the occurrence and genetic identity of Piroplasmida agents in this species is still cloudy. Although it is reported that Theileria equi, an endemic equid-infective agent in Brazil, is occurring in lowland tapirs, these reports are probably misconceived diagnoses since they are solely based on small fragments of 18S rRNA that may not achieve accurate topologies on phylogenetic analyses. The present study aimed to detect and investigate the identity of Theileria spp. in tapirs from Pantanal and Cerrado biomes. Blood-DNA samples from tapirs were screened for a partial (~800 bp) 18S rRNA gene fragment from Piroplasmida and 64 (64/122; 52.46% CI: 43.66–61.11%) presented bands of expected size. Samples were submitted to different protocols for molecular characterization, including near-full length 18S rRNA gene (~1500 bp), and the ema-1 gene from T. equi. Eight sequences were obtained for extended fragments (1182–1473 bp) from the 18S rRNA gene. Moreover, three sequences from partial cox-1 and five from partial hsp70 gene were obtained. None of the samples presented amplifications for the ema-1 gene. Phylogenetic and distance analyses from the 18S rRNA sequences obtained demonstrated a clear separation from tapirs’ Theileria spp. and T. equi. Phylogenetic analyses of cox-1 and hsp70 sequences obtained herein also showed a unique clade formed by tapir’s Theileria spp. Theileria terrestris sp. nov. is positioned apart from all other Theileria species in 18S rRNA, cox-1, and hps70 phylogenetic analyses. This novel proposed species represents a new Piroplasmida clade, yet to be characterized regarding biological features, vectors involved in the transmission cycles, additional vertebrate hosts, and pathogenicity

Neorickettsia sp. in coatis (Nasua nasua) in Brazil

Abstract 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

Pathological findings associated with Dipetalonema spp. (Spirurida, Onchocercidae) infection in two species of Neotropical monkeys from Brazil

Among vector-borne helminths, filarioids of the genus Dipetalonema (Spirurida: Onchocercidae) localize in several tissues and body cavities of several animal species, causing mild to moderate lesions. The pathological findings associated with Dipetalonema spp. infection in Neotropical monkeys from southern Brazil are herein described, along with a fatal case due to filarial polyserositis and entrapment of an intestinal segment. At necropsy, nematodes were observed in abdominal and thoracic cavities, or in the pericardium of 37 (31.3%) out of the 118 individuals examined (i.e., 35 Alouatta guariba clamitans and two Sapajus nigritus). In addition, at histology, 27.0% of positive animals presented microfilarie (inside blood vessels of lung, spleen, liver, and brain) and 8.1% presented adult nematodes in the heart, lung, and liver. In two cases, cross-sections of filarioids were associated with areas of epicardial thickening with intense fibrosis and pyogranulomatous inflammation in the brain, heart, liver, lungs, or spleen. The DNA fragment was amplify using the cox1 gene, sequenced and analyzed to identify the nematode species collected; presence of Wolbachia was assessed in the filarioids using the 16S rRNA gene. At BLAST analysis of the cox1 gene, 10 sequences showed 91.7% nucleotide identity with Dipetalonema gracile, and two with D. gracile (98.5%) and Dipetalonema graciliformis (98.3%). Phylogenetic analyses clustered sequences of the cox1 obtained in this study in two clades corresponding with the host species. Wolbachia sp. endosymbiont was detected in four samples. Data herein reported provide a description of pathological lesions associated with the infection by Dipetalonema spp., suggesting that they may cause disease in Neotropical monkeys. In addition, a better understanding of diversity and biology of Dipetalonema spp. in South America is needed to assess the impact they may cause in native non-human primates from Brazil

Multi-Locus Sequencing Reveals Putative Novel Anaplasmataceae Agents, ‘<i>Candidatus</i> Ehrlichia dumleri’ and <i>Anaplasma</i> sp., in Ring-Tailed Coatis (Carnivora: <i>Nasua nasua</i>) from Urban Forested Fragments at Midwestern Brazil

The Anaplasmataceae family encompasses obligate intracellular α-proteobacteria of human and veterinary medicine importance. This study performed multi-locus sequencing to characterize Ehrlichia and Anaplasma in coati’s blood samples in Midwestern Brazil. Twenty-five samples (25/165—15.1%) were positive in the screening PCR based on the dsb gene of Ehrlichia spp. and were characterized using 16S rRNA, sodB, groEL, and gltA genes and the 23S-5S intergenic space region (ITS). Phylogenetic analyses based on all six molecular markers positioned the sequences into a new clade, with a common origin of Ehrlichia ruminantium. Haplotype analyses of 16S RNA sequences revealed the presence of two distinct Ehrlichia genotypes. Six samples (6/165, 3.6%) were positive in the screening nPCR for the 16S rRNA gene of Anaplasma spp. and were submitted to an additional PCR targeting the ITS for molecular characterization. Phylogenetic analyses based on both 16S rRNA gene and ITS positioned the Anaplasma sp. detected in the present study in a large clade with other Anaplasma sp. previously detected in ticks and wild animals and in a clade with ‘Candidatus Anaplasma brasiliensis’, respectively. Based on distinct molecular markers, the present work described a putative novel Anaplasmataceae agent, namely ‘Candidatus Ehrlichia dumleri’, and Anaplasma sp. closely related to the previously described ‘Candidatus Anaplasma brasiliensis’

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.Discussion: Preoperative planning for orthopedic surgery in rabbits is different from that of dogs and cats, due to the particularities of the species. Familiarity with the regional anatomy, patient preparation, and appropriate instrumentation are necessary when contemplating osteosynthesis in a rabbit. Salter Harris fractures affect young animals, where the physis is considered an area of fragility in the bone. Surgical planning should take into account the function of the growth plates.It is recommended that implants passing through the physis do so perpendicularly, since angulation greater than 45° may predispose the bone to premature closure of the growth plate. Another important consideration involves the choice of implants, since the use of trocar-tipped pins facilitates their precise placement in the bone. The use of threaded pins should be avoided due to their weakness at the thread-shaft interface, and the risk of impaired longitudinal bone growth and thedifficulty of removal if necessary. Steinman pins and Kirschner wires can be used to stabilize a variety of different fractures. In Salter-Harris type I and II fractures, the use of pins neutralizes bending forces but not rotational or compressive forces. In contrast, Rush pinning and cross-pinning techniques are effective in neutralizing the forces acting on the physis, and are frequently used for the fixation of fractures in this region. Complications, often associated with poor reduction and alignment, can result in varus or valgus deviation and are associated with a high risk of implant failure or migration, malunion, and patellar luxation. It was concluded that the combination of Rush pins and an intramedullary pin resulted in adequate stabilization of the Salter Harris type I fracture of the distal femur in this rabbit.Keywords: Orthopedic implants, osteosynthesis, trauma, bone repairing, lagomorph

Molecular Survey of Hemotropic <i>Mycoplasma</i> spp. and <i>Bartonella</i> spp. in Coatis (<i>Nasua nasua</i>) from Central-Western Brazil

Even though previous works showed molecular evidence of hemotropic Mycoplasma spp. (hemoplasmas) in ring-tailed coatis (Nasua nasua) from Brazil, Bartonella sp. has not been reported in these mammals so far. The present study aimed to detect the above-mentioned agents in coatis’ blood and associated ectoparasites, assessing the association between these infections and red blood parameters. Between March 2018 and January 2019, coati (n = 97) blood samples, Amblyomma sp. ticks (2242 individual ticks, resulting in 265 pools), and Neotrichodectes pallidus louse (n = 59) were collected in forested urban areas from midwestern Brazil. DNA extracted from coatis’ blood, and ectoparasite samples were submitted to quantitative PCR (qPCR) (16S rRNA) and conventional PCR (cPCR) (16S rRNA and 23S rRNA) for hemoplasmas and qPCR (nuoG gene) and culturing (only blood) for Bartonella spp. Two different hemoplasma genotypes were detected in blood samples: 71% coatis positive for myc1 and 17% positive for myc2. While 10% of ticks were positive for hemoplasmas (myc1), no louse was positive. The estimated bacterial load of hemoplasmas showed no association with anemia indicators. All coatis were negative for Bartonella sp. in qPCR assay and culturing, albeit two Amblyomma sp. larvae pools, and 2 A. dubitatum nymph pools were positive in the qPCR. The present work showed a high occurrence of hemoplasmas, with two distinct hemoplasma genotypes, in coatis from forested urban areas in midwestern Brazil

Parasitic fauna of bats from Costa Rica

Bats are important reservoirs and spreaders of pathogens, including those of zoonotic concern. Though Costa Rica hosts one of the highest bat species' diversity, no information is available about their parasites. In order to investigate the occurrence of vector-borne pathogens (VBPs) and gastrointestinal (GI) parasites of chiropterans from this neotropical area, ectoparasites (n = 231) and stools (n = 64) were collected from 113 bats sampled in Santa Cruz (site 1) and Talamanca (site 2). Mites, fleas and ticks were morphologically and molecularly identified, as well as pathogens transmitted by vectors (VBPs, i.e., Borrelia spp., Rickettsia spp., Bartonella spp.) and from feces, such as Giardia spp., Cryptosporidium spp. and Eimeria spp. were molecularly investigated. Overall, 21 bat species belonging to 15 genera and 5 families were identified of which 42.5% were infested by ectoparasites, with a higher percentage of mites (38.9%, i.e., Cameronieta sp. and Mitonyssoides sp.) followed by flies (2.6%, i.e., Joblingia sp.) and tick larvae (1.7%, i.e., Ornithodoros sp.). Rickettsia spp. was identified in one immature tick and phylogenetically clustered with two Rickettsia species of the Spotted Fever Group (i.e., R. massiliae and R. rhipicephali). The frequency of GI parasite infection was 14%, being 3.1% of bats infected by Giardia spp. (un-identified non-duodenalis species), 1.5% by Eimeria spp. and 9.4% by Cryptosporidium spp. (bat and rodent genotypes; one C. parvum-related human genotype). The wide range of ectoparasites collected coupled with the detection of Rickettsia sp., Giardia and Cryptosporidium in bats from Costa Rica highlight the role these mammals may play as spreaders of pathogens and the need to further investigate the pathogenic potential of these parasites