Investigação de Salmonella spp. em Psittaciformes exóticos e nativos mantidos em cativeiro na região central do Rio Grande do Sul / Investigation of Salmonella spp. in exotic and native Psittaciformes kept in captivity in the central region of Rio Grande do Sul

As doenças infecciosas são reconhecidamente causadoras de declínios populacionais de animais silvestres e algumas delas podem representar ameaça à saúde pública. O presente estudo objetivou investigar a ocorrência de Salmonella spp. em Psittaciformes exóticos e nativos mantidos em cativeiro na região central do Rio Grande do Sul, além de comparar os resultados obtidos por meio do método bacteriológico convencional e da reação em cadeia da polimerase (PCR), mediante a utilização de material fecal do ambiente, evitando-se o estresse da contenção. Durante os meses de agosto/2016 e setembro/2016 foram coletadas amostras de fezes frescas de 90 gaiolas em dois criatórios, representando 180 aves. Com as duas técnicas empregadas nas análises, as amostras foram negativas para Salmonella spp., contudo foram detectadas bactérias da família Enterobacteriaceae: Escherichia coli, Cedecea sp. e Citrobacter freundii. Considera-se importante a continuidade do monitoramento dos criatórios, já que o patógeno investigado pode estar em aves portadoras e se manifestar em situações de estresse, representando riscos enquanto zoonose e prejuízos à saúde das aves.(AU)

Infectious diseases are known to cause decline in wildlife population and some of them may represent a threat to public health. This study aimed to investigate the occurrence of Salmonella spp. in exotic and native Psittaciformes kept in captivity in the central region of Rio Grande do Sul, Brazil, and to comparing the results obtained through the conventional bacteriological method and the polymerase chain reaction (PCR), using fecal matter collected from the environment, avoiding the containment stress. From August to September 2016, samples of fresh feces were collected from 90 cages in two breeding grounds, representing 180 birds. According to both analysis techniques the samples were negative for Salmonella spp., however, bacteria from the Enterobacteriaceae family: Escherichia coli, Cedecea sp. and Citrobacter freundii were detected. Continuous monitoring of breeding grounds is important since the pathogen investigated can be found in carrier birds and manifest itself in stressful situations presenting risks such as zoonosis and reduced bird health.(AU)

Piometra em uma leoa (Panthera leo): relato de caso / Pyometra in a lion (Panthera leo): case report

A piometra é uma infecção aguda ou crônica do útero que ocorre frequentemente em cadelas não castradas, podendo também ocorrer em gatas domésticas e selvagens, sendo poucos os estudos relacionados à piometra em grandes felídeos. O objetivo deste relato foi descrever um caso de piometra em uma leoa (Panthera leo) de cativeiro, as lesões de necropsia e histológicas, bem como os resultados da análise microbiológica. Uma leoa com aproximadamente 23 anos, pertencente a um criadouro conservacionista de Santa Maria-RS, foi encontrada morta pela manhã em seu recinto. Após coleta de dados, procedeu-se à necropsia e à coleta de material para análise histopatológica e bacteriológica. A análise microbiológica revelou predomínio das bactérias Streptococcus sp. e Escherichia coli no conteúdo purulento do útero, caracterizando como piometra, e a bactéria predominante em plasma, fígado e medula óssea foi E. coli. De acordo com o laudo histopatológico, as alterações observadas nessa leoa sugerem um quadro de septicemia grave, sendo a origem do foco infeccioso bacteriano, provavelmente, a piometra. Considera-se importante chamar a atenção dos médicos veterinários de animais selvagens para um diagnóstico precoce dessa doença, que é comum em cadelas, mas que pode acometer também felídeos selvagens e levá-los à morte.

Pyometra is an acute or chronic uterus infection that occurs often in not spayed dogs, but may also occur in wild and domestic cats, and there are few studies related to pyometra in big cats. The aim of this report was to describe a case of pyometra in a lion (Panthera leo) in captivity, the gross and microscopic lesions found at necropsy, and the results of the microbiological analysis. A female with approximately 23 years of age, belonging to a Conservationist Breeding Center located in Santa Maria-RS-Brazil was found dead in her enclosure in the morning. After data collection, we proceeded to the autopsy and collection of material for histopathological and bacteriological analyzes. Microbiological analysis revealed a predominance of the bacteria Streptococcus sp and Escherichia coli in the uterus content, characterized as pyometra and the predominant bacterium in the plasma, liver and bone marrow was Escherichia coli. According to histopathology, the changes observed in this lioness suggest a framework of severe septicemia, being the source of the bacterial infection, probably, pyometra. It is considered important to draw the attention of zoo and wildlife veterinarians for an early diagnosis of this common disease in dogs, which can also affect large wild cats and lead them to death.

Atividade de nanoformulações de Melaleuca alternifolia e terpinen-4-ol em isolados de Rhodococcus equi / Melaleuca alternifolia activity in nanoformulations and terpinen-4-ol against Rhodococcus equi isolates

Rhodococcus equi é o agente etiológico da rodococose equina, importante doença respiratória de potros. Especialmente na última década, a emergência de cepas resistentes aos antimicrobianos empregados no tratamento da rodococose tem sido relatada. Nesse sentido, há a necessidade de estudos envolvendo terapias alternativas e novas tecnologias, incluindo o uso de plantas medicinais e nanotecnologia. Neste trabalho utilizou-se Melaleuca alternifolia nas seguintes formulações: óleo livre, nanocápsula, nanoemulsão e a combinação de óleo livre com nanocápsula e com nanoemulsão, além do seu composto majoritário, terpinen-4-ol, a fim de verificar a atividade antimicrobiana frente a isolados de R. equi de diferentes origens. Utilizou-se o método de microdiluição em caldo na determinação das concentrações inibitória mínima (CIM) e bactericida mínima (CBM) das diferentes formulações frente aos isolados (n=24). Verificou-se baixo potencial para atividade antibacteriana de M. alternifolia na formulação de óleo livre. Todavia, essa atividade foi potencializada quando se incorporou o óleo essencial às nanoformulações. O composto terpinen-4-ol demonstrou potencial atividade antibacteriana quando incorporado ao óleo essencial e quando utilizado isoladamente. Verificou-se que tanto M. alternifolia quanto terpinen-4-ol testados possuem atividade antimicrobiana contra isolados de R. equi, sugerindo seu emprego em estudos avaliando seu potencial para o tratamento da rodococose.

Rhodococcus equi causes rodococose in horses, characterized by bronchopneumonia in foals. Due to reports of antimicrobial resistance, it is important to develop studies involving alternative therapies and new technologies, including the use of medicinal plants and nanotechnology. In this work, the plant Melaleuca alternifolia in oil free formulations, nanocapsule, nanoemulsion and the combination of free and nanocapsule oil nanoemulsion, besides its major compound, terpinen-4-ol, were used in order to verify antimicrobial activity against isolates of R. equi. The broth microdilution method was employed to determine the minimum inhibitory (MIC) and minimum bactericidal (MBC) concentrations of different formulations against 24 isolates. There was low antibacterial activity of M. alternifolia in oil free formulation; however, it was observed that the activity was enhanced when incorporated as essential oil the nanoformulations. The major compound, terpinen-4-ol, showed bactericidal and bacteriostatic activity when used alone. It is suggested that M. alternifolia, in association with nanocarriers systems, as well as terpinen -4-ol, presents potential for future studies concerning the equine rodococosis therapy.

Contrasting genetic diversity and differentiation of populations of two successional stages in a neotropical pioneer tree (Eremanthus erythropappus, Asteraceae)

Eremanthus erythropappus, commonly known as “candeia”, is an abundant pioneer tree species, forming dense populations known as “candeial”, but it is also found in forests at middle stages of succession. Trees from forests are bigger and occur in lower density than in the “candeial”. The objectives of the present study were to investigate if the decrease in population density during successional process is accompanied by 1) changes in within-population genetic diversity, and 2) differentiation of populations. Eight populations, four of early successional stage (“candeial”) and four of middle successional stages (forest), were analyzed with RAPD markers. The genetic diversity found was high compared to other tree species analyzed with RAPD markers. AMOVA revealed that most of the genetic variations of E. erythropappus were found within populations (85.7%), suggesting that this species is predominantly outcrossing. The relatively low differentiation among the populations can be attributed to small distances among the populations analyzed (0.2 to 10.8 km). No indication that populations from middle successional habitats show lower genetic variation than populations from early successional stages was found. The percentage of polymorphic fragments (82.8 and 84.8%) and the Shannon indexes (0.442 and 0.455) were similar in “candeial” and forest, respectively. These results suggest that if an increase in selection intensity occurred during succession, it did not result in a decrease in genetic diversity or that the selection effect was balanced by other factors, such as gene flow. Higher significant differentiation among E. erythropappus populations from “candeial” in relation to that among populations from forest was also not detected.

Comparative analysis of different DNA extraction protocols in fresh and herbarium specimens of the genus Dalbergia

Five published DNA extraction protocols were compared for their ability to produce good quality DNA from fresh and herbarium leaves of several species of the genus Dalbergia. The leaves of these species contain high amounts of secondary metabolites, which make it difficult to perform a clean DNA extraction and thereby interfering with subsequent PCR amplification. The protocol that produced the best DNA quality in most of the Dalbergia species analyzed, utilizes polyvinylpyrrolidone to bind the phenolic compounds, a high molar concentration of NaCl to inhibit co-precipitation of polysaccharides and DNA, and LiCl for removing RNA by selective precipitation. The DNA quality of herbarium specimens was worse than that for fresh leaves, due to collecting conditions and preservation of samples. We analyzed 54 herbarium specimens, but the recovered DNA allowed successful PCR amplification in only eight. For the genus Dalbergia, the herbarium is an important source of material for phylogenetic and evolutionary studies; due to the occurrence of the different species in various geographical regions in Brazil, it is difficult to obtain fresh material in nature. Our results demonstrated that for Dalbergia species the methods used for the collection and preservation of herbarium specimens have a mayor influence on DNA quality and in the success of phylogenetic studies of the species

Applicability of RAPD markers on silver-stained polyacrylamide gels to ascertain genetic diversity in Peripatus acacioi (Peripatidae; Onychophora)

RAPD (random amplification of polymorphic DNA) molecular markers can be utilized for analyzing genetic variability in populations for which only a few or no molecular markers are available. They were used in a study of an endangered species, Peripatus acacioi, found in the Tripuí Ecological Station, in Ouro Preto, MG, Brazil. The ecological station was specifically created to protect this velvet worm species, the first of this group found in Brazil. For an initial evaluation of the genetic diversity of this species, DNA samples from the lobopods of four individuals, collected at random, were analyzed using RAPD. Each reaction was run with a different primer (Operon RAPD 10-mer Kits), totaling 13 primers (OPC2, OPC3, OPC4, OPC6, OPC8, OPC10, OPC11, OPL2, OPL7, OPL11, OPL13, OPL18, and OPL19). Due to the low amplification yield, RAPD fragments were separated in polyacrylamide gels and stained with silver nitrate. Numerous bands were observed. Fifty-five of the amplified bands proved to be reproducible, both in terms of presence and intensity. Among these, 27 were variable and 28 were constant. The average number of bands per gel was 4.2. Nine of the 13 primers tested allowed the identification of constant and variable bands among these four individuals. RAPD analysis of genetic variation using silver-stained polyacrylamide gel electrophoresis provided measures of band sharing among the individuals, and therefore could be used in population genetics studies of P. acacioi.