High-resolution melting (HRM) analysis is a very attractive and flexible advanced post-PCR method with high sensitivity/specificity for simple, fast and cost-effective genotyping based on the detection of specific melting profiles of PCR products. Next generation real-time PCR systems, along with improved saturating DNA-binding dyes, enable the direct acquisition of HRM data after quantitative PCR. Melting behaviour is particularly influenced by the length, nucleotide sequence and GC content of the amplicons. This method is expanding rapidly in several research areas such as human genetics, reproductive biology, microbiology and ecology/conservation of wild populations. Here we have developed a successful HRM protocol for avian sex identification based on the amplification of sex-specific CHD1 fragments. The melting curve patterns allowed efficient sexual differentiation of 111 samples analysed (plucked feathers, muscle tissues, blood and oral cavity epithelial cells) of 14 bird species. In addition, we sequenced the amplified regions of the CHD1 gene and demonstrated the usefulness of this strategy for the genotype discrimination of various amplicons (CHD1Z and CHD1W), which have small size differences, ranging from 2 bp to 44 bp. The established methodology clearly revealed the advantages (e.g. closed-tube system, high sensitivity and rapidity) of a simple HRM assay for accurate sex differentiation of the species under study. The requirements, strengths and limitations of the method are addressed to provide a simple guide for its application in the field of molecular sexing of birds. The high sensitivity and resolution relative to previous real-time PCR methods makes HRM analysis an excellent approach for improving advanced molecular methods for bird sexing.
This study aimed to analyze the admission causes, outcomes, primary causes of death, and main lesions observed in the post mortem examinations of Western European hedgehogs, Erinaceus europaeus (Linnaeus, 1758), in the north of Portugal. The data were obtained by consulting the records from the two main wildlife rehabilitation centers located in the north of Portugal (Wildlife Rehabilitation Centre of Parque Biologico de Gaia and the Wildlife Rehabilitation Centre of the University of Trás-os-Montes and Alto Douro). Over 17 years (2002–2019) a total of 740 animals were admitted. Most of the animals were juveniles, with the highest number of admissions occurring during summer (36.8%) and spring (33.2%). The main reasons for admission were debilitation (30.7%) and random finds (28.4%). Of the total number of individuals admitted to these centers, 66.6% were successfully released back into the wild. The most relevant causes of death were trauma of unknown origin (32.7%), nontrauma causes of unknown origin (26.6%), and nutritional disorders (20.2%). The main lesions observed were related to trauma, including skeletal and skin lesions (fractures, hemorrhages, wounds) and organ damage, particularly to the lungs and liver. The hedgehog is a highly resilient and adaptable animal. The urban environment has many benefits for hedgehogs, yet the presence of humans can be harmful. In the future, the public needs to become even more involved in the activities of the wildlife centres, which will make a positive difference for these populations.
Background-Straelensia cynotis, a trombidioid larval mite, was identified as a cause of nodular dermatitis in dogs in Southern Europe. It has been suggested that red fox (Vulpes vulpes) is a natural host for S. cynotis. However, no case has been reported in this species. Objective-To describe three suspected cases of straelensiosis in red foxes. Animals-Three juvenile wild red foxes from Portugal. Results-Erythematous papules and nodules were found in the head, neck and limbs of these foxes with no associated pruritus. In skin biopsies, well-preserved larval mites were found within dilated hair follicles. These follicular lesions were multifocal and consisted of a degenerative and necrotic area nearby the parasite's mouthparts with marked pseudoepitheliomatous hyperplasia and perifollicular mucinosis. These features are considered pathognomonic in S. cynotis infestations in dogs. Treatment and outcome are outlined. Conclusions and clinical importance-To the best of the authors' knowledge, the present case series documents the first known occurrence of nodular dermatitis by Straelensia spp. in red foxes. This new evidence may corroborate participation of the red fox in the life cycle of S. cynotis.
RESUMO.-[Osteossíntese de fratura localizada no membro torácico num Lobo Ibérico (Canis lupus signatus) selvagem e sua reabilitação.] O objetivo do presente artigo é a descrição da realização da estabilização de uma fratura cominutiva exposta do rádio e ulna do membro torácico direito em Lobo Ibérico selvagem por meio de osteossíntese com placa e parafusos ósseos bem como o manejo pós-operatório. A evolução da cicatrização da fratura óssea foi similar à esperada num canídeo doméstico de porte idêntico. A intervenção cirúrgica realizada rapidamente, bem como o alojamento adequado, alimentação e manejo adaptado a um lobo selvagem, visando minimizar a manipulação e o aparecimento de possíveis complicações pós-operatórias permitiram o sucesso na reabilitação e posterior libertação do animal. Na 10ª semana pós-operatória foi colocado um colar GPS no animal para deteção permanente da sua localização, tendo sido o animal libertado na mesma área onde havia sido capturado. The surgical treatment of an exposed compounded comminuted fracture of the right radius and ulna in a free-ranging adult female Iberian Wolf (Canis lupus signatus) with an osteosynthesis plate and screws and subsequent post-operative care are described. The evolution of the fracture healing was very similar to those expected in a dog of the same size. The prompt surgical intervention and a proper housing, feeding and wound management adapted to a free-ranging wolf, in view to reduce manipulation and post-operative complications, allowed the subsequent rehabilitation and release of the animal. After 10 th post-operative weeks the wolf was fitted with a Global Positioning System (GPS) for wildlife tracking collar and released in the same area where it has been caught. GPS telemetry data showed that the animal covered increasingly large distances confirming a complete functionality of the right thoracic limb and its successfully return to the wild. This report could constitute the first detailed report of a long bone fracture treatment in a free-ranging wolf and its successfully rehabilitation, release and adaptation to the wild. recuperação do membro torácico direito e o sucesso da sua reintrodução na natureza. Este artigo poderá constituir a primeira descrição pormenorizada sobre a estabilização de fratura de ossos longos em um lobo selvagem com a sua posterior reintrodução na vida selvagem com sucesso.
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