A genetic and virulence characterization of Brazilian strains of Mycoplasma hyopneumoniae

Mycoplasma hyopneumoniae (M. hyopneumoniae) is considered the primary causative agent of porcine enzootic pneumonia (EP), a chronic contagious respiratory disease that causes economic losses. Obtaining new pathogenic isolates and studying the genome and virulence factors are necessary. This study performed a complete sequencing analysis of two Brazilian strains, UFV01 and UFV02, aiming to characterize the isolates in terms of the virulence factors and sequence type. The complete genome analysis revealed the main virulence genes (mhp385, mhp271, MHP_RS03455, p102, p97, p216, MHP_RS00555, mhp107) and ST-123, the presence of three toxin-related genes (tlyC, PLDc_2 and hcnC), and some genetic groups specific to these two isolates. Subsequently, the pathogenicity of the isolates was evaluated via an experimental infection conducted in a swine model. The study was divided into three groups, namely a negative control group (n = 4) and two test groups (n = 8), totaling 20 animals. They were challenged at 35 days of age with 107 CCU (Color Changing Units) M. hyopneumoniae via the intratracheal route. The UFV01 group showed earlier and higher seroconversion (IgG) (100%), while only 50% of the UFV02 group seroconverted. The same trend was observed when analyzing the presence of IgA in the bronchoalveolar lavage fluid (BALF) at 35 days post-infection (dpi). The UFV01 group had a mean macroscopic lesion score of 11.75% at 35 dpi, while UFV02 had 3.125%. Microscopic lesions were more severe in the UFV01 group. Based on laryngeal swab samples evaluated by qPCR, and the detection began at 14 days. The UFV01 group showed 75% positivity at 14 dpi. The UFV02 group also started excreting at 14 dpi, with a positivity rate of 37.5%. The results indicate that the UFV01 isolate exhibits higher virulence than UFV02. These findings may aid in developing new vaccines and diagnostic kits and establishing experimental models for testing

Records of ectoparasites on humans and wildlife in southeastern Brazil

We report ectoparasites found on humans and wildlife from different locations in southeastern Brazil. Ectoparasites were stored in 70% alcohol and visually identified under a stereo microscope with the help of identification keys. Genomic DNA of ticks was extracted and used in the Polymerase Chain Reaction (PCR) to detect the presence of tick-borne pathogens. Sixteen species of ectoparasites (six ticks, one lice, four fleas, four flies, and one bedbug) were identified on ten host species, including humans, other mammals, and birds. We report the first record of the parasitism of Amblyomma longirostre on Didelphis aurita, Microlynchia pussila on Turdus leucomelas, Ornithoica vicina and Ornithocoris pallidus on Psittacara leucophthalmus and O. pallidus on Homo sapiens. In addition, we also report the first record of O. vicina in the state of Minas Gerais and the infection by Rickettsia sp. in Amblyomma varium

Salmonella spp. in poultry carcass: evaluation of sample preparation methods and effect of storage under refrigeration on pathogen recovery

The aim of the present study was to evaluate the use of different analytical units and the influence of storage under refrigeration on the detection of Salmonella sp. in naturally contaminated poultry carcasses. One hundred and thirty samples were collected during the production process soon after chilling (postchiller phase). Fifty-five samples were analyzed in up to 2 h after collection and 65 samples were analyzed after 72 h of storage. Pathogen screening was based on three different analytical units and a comparison was made between them. Carcasses were initially rinsed with 400 mL of diluent, and three different analytical units were incubated: total rinsing volume (TRV), a single 30 mL aliquot of the rinsing volume, and 25 g of skin from different areas of the carcass. Of all samples analyzed, 60% were positive for Salmonella sp. From the samples collected at the post-chiller phase, 57% were positive for the pathogen and 52.31% of these were detected by TRV; a better statistical performance (P0.05). There were no significant differences between the number of positive samples from the post-chiller phase and after 72 h of refrigeration. It was also seen that the use of different analytical units (one for the post-chiller phase and another for the refrigerated samples) in samples coming from the same production lot may give different results.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Histologia da associação de membranas biológicas de origem bovina implantadas no subcutâneo de ratos

O trabalho teve por objetivo avaliar a capacidade de barreira da membrana de cortical óssea desmineralizada liofilizada bovina, testando sua permeabilidade e integridade, em associação com a membrana de pericárdio bovino liofilizado. Em 15 ratos Wistar, machos, adultos, implantou-se no subcutâneo da região cérvico-torácica dorsal a combinação de duas membranas de cortical óssea desmineralizada tendo no interior a membrana de pericárdio bovino. Os animais (cinco ratos/grupo) foram submetidos à eutanásia aos 15, 30 e 60 dias de pós-operatório. A avaliação microscópica mostrou que, aos 15 dias de pós-cirúrgico, as membranas estavam íntegras, com exceção de pequenas áreas de reabsorção da membrana de cortical óssea junto aos locais referentes aos antigos canais nutritivos. Aos 30 dias, havia apenas restos da membrana de cortical óssea e, aos 60 dias, a sua total ausência. Baseados nos resultados aqui obtidos, é possível concluir que a membrana de cortical óssea bovina é rapidamente absorvida e não confere proteção à membrana de pericárdio bovino liofilizado

Histologia da associação de membranas biológicas de origem bovina implantadas no subcutâneo de ratos

O trabalho teve por objetivo avaliar a capacidade de barreira da membrana de cortical óssea desmineralizada liofilizada bovina, testando sua permeabilidade e integridade, em associação com a membrana de pericárdio bovino liofilizado. em 15 ratos Wistar, machos, adultos, implantou-se no subcutâneo da região cérvico-torácica dorsal a combinação de duas membranas de cortical óssea desmineralizada tendo no interior a membrana de pericárdio bovino. Os animais (cinco ratos/grupo) foram submetidos à eutanásia aos 15, 30 e 60 dias de pós-operatório. A avaliação microscópica mostrou que, aos 15 dias de pós-cirúrgico, as membranas estavam íntegras, com exceção de pequenas áreas de reabsorção da membrana de cortical óssea junto aos locais referentes aos antigos canais nutritivos. Aos 30 dias, havia apenas restos da membrana de cortical óssea e, aos 60 dias, a sua total ausência. Baseados nos resultados aqui obtidos, é possível concluir que a membrana de cortical óssea bovina é rapidamente absorvida e não confere proteção à membrana de pericárdio bovino liofilizado.The aim of this study was to evaluate the barrier capacity of the bovine demineralized lyophilized bone cortical membrane, testing its permeability and its integrity in association with lyophilized bovine pericardium membrane. Fifteen male adult Wistar rats were used. The combination of two bone cortical membranes with bovine pericardium membrane inside was implanted subcutaneously in the cervical-thoracic area. The animals (five rats per group) were submitted to euthanasia at 15, 30 and 60 days postoperatively. on day 15 postoperative, the microscopic evaluation showed integrity of the membranes, except the absorption of small areas of the bone cortical membrane in the sites of nutritive channels. on day 30, there was a remnant of bone cortical membrane, and on day 60 bone cortical membrane was completely absent. It was possible to conclude that bovine demineralized lyophilized bone cortical membrane is quickly absorbed and does not preserve the lyophilized bovine pericardium membrane

Mercury concentrations in south atlantic swordfish, Xiphias gladius, Caught off the Coast of Brazil

This study evaluated the concentrations of mercury in fillets (anterior, middle, and end regions) from the swordfish, Xiphias gladius, and the relationships between mercury concentration and fish weight, as well as the region of collection. Of a total of 697 swordfish analyzed, 11 had mercury concentrations above 1 mg/kg, 421 were between 0.5 and 1.0 mg/kg, and 265 were below 0.5 mg/kg. The anterior and posterior regions had greater concentrations of mercury than the middle region, and fish caught off the northern coast of Brazil had a higher concentration than those caught off the southern coast. © 2013 Springer Science+Business Media New York

Genomic Analyses of Weissella cibaria W25, a Potential Bacteriocin-Producing Strain Isolated from Pasture in Campos das Vertentes, Minas Gerais, Brazil

Weissella is a genus containing Gram-positive, heterofermentative bacteria belonging to the lactic acid bacteria (LAB) group. These bacteria are endowed with promising technological and antimicrobial attributes. Weissella cibaria W25 was isolated from a dairy environment where raw milk cheeses are produced. Therefore, we sequenced and assembled the W25 draft genome sequence, which consists of 41 contigs totaling ~2.4 Mbp, with a G + C content of 45.04%. Then we carried out a comprehensive comparative genomic analysis with W. cibaria 110, known to produce the weissellicin 110 bacteriocin, and four other non-bacteriocin-producing W. cibaria strains

A refrigeração no diagnóstico de Salmonella spp. utilizando o método microbiológico tradicional e reação em cadeia da polimerase em carcaças de frango

A pesquisa de Salmonella em carcaças de aves tem mostrado resultados discrepantes, dependendo se as amostras foram colhidas ainda na indústria, imediatamente após o chiller ou no comércio varejista, quando se encontram submetidas à refrigeração por vários dias. Técnicas mais sensíveis, tais como a Reação em Cadeia pela Polimerase (PCR), podem fornecer dados importantes sobre o efeito do resfriamento sobre as células do patógeno, comparando seus resultados com os da metodologia microbiológica convencional (MC). Foram colhidas 130 carcaças de frango, sendo que 65 foram obtidas ainda na indústria (pós-chiller) e imediatamente analisadas e 65 carcaças embaladas e estocadas a 5°C por 72 horas (simulando o varejo), sendo então realizada a pesquisa do patógeno por ambas as técnicas. do total analisado (130 amostras), a PCR foi capaz de detectar 58 positivas (44,6%) e a MC 50 (38,5%). Ambas as técnicas detectaram um número superior de amostras positivas para Salmonella em carcaças colhidas ainda na indústria, quando comparadas às do varejo. A PCR detectou 50,77% de positividade em amostras da indústria e 38,46% em amostras do varejo. Para a MC, esses valores foram de 46,15% (indústria) e 30,77% (varejo). Concluímos que o resfriamento das carcaças a 5°C por 72 horas pode ser um fator limitante na detecção de Salmonella quando a pesquisa do patógeno se faz pela metodologia microbiológica convencional.The survey of Salmonella in poultry carcasses has shown conflicting results, depending on whether the samples were taken yet in the factory, immediately after the chiller or retail market, when they are subjected to refrigeration for several days. More sensitive techniques such as Polymerase Chain Reaction (PCR) can provide important data on the effect of cooling on the cells of the pathogen by comparing their results with those of conventional microbiological methods (CM). It was collected 130 chicken carcasses, wich 65 were obtained yet in the industry (post-chiller) and immediately analyzed and 65 carcasses packaged and stored at 5°C for 72 hours (simulating retail), and then research of the pathogen was performed by both techniques. of the analyzed total (130 samples), PCR was able to detect 58 positive (44.6%) and CM, 50 (38.5%). Both techniques detected a higher number of samples positive for Salmonella on carcasses collected yet in the factory when compared to those of retail. The PCR detected 50.77% of positive industry samples and 38.46% of retail samples. CM for these values was 46.15% (factory) and 30.77% (retail). We conclude that the cooling of the carcasses for at 5°C for 72 hours can be a limiting factor in the detection of Salmonella when the pathogen research is done by conventional microbiological methods.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Avaliação da unidade analítica na detecção de Salmonella spp. em frangos varejo

Over 95 % of the cases of human salmonellosis are foodborne. Among the foods involved in transmission of these diseases are the products from animal origin, being the poultry products highly relevant. To deal with this issue, the Ministry of Agriculture, Livestocks Farming and Supply (MAPA – Ministério da Agricultura, Pecuária e Abastecimento) established a program, in order to monitor the contamination by Salmonella spp. in poultry carcasses. This study aimed at comparing the efficiency of two analytical methodologies (rinsing of chicken carcass and skin excision) for recovering the cells from this bacterium and to verify the occurrence of this pathogen in samples collected in Botucatu, SP. Analyzing 100 carcasses, and computing the results by both techniques, contamination by Salmonella spp. was detected in 43% samples. Using the rinsing technique, 81.40% (35/43) showed positive results as well as 51.16% (22/43) using the skin excision methodology. Although the rinsing technique detected high number of positive samples, no statistical difference (p > 0.05) was found between the results obtained from two methodologies. Further studies should be conducted on this issue as the poultry carcasses are crucial source for transmission of Salmonella spp., and the subsequent introduction of this pathogen on food preparation environment.Mais de 95% das salmoneloses humanas são de origem alimentar e, entre os alimentos envolvidos na transmissão estão os produtos de origem animal, especialmente os de maior relevância, os produtos avícolas. Diante desta questão, o Ministério de Agricultura, Pecuária e Abastecimento (MAPA) estabeleceu um programa com a finalidade de reduzir e monitorar a contaminação por Salmonella spp. nas carcaças de aves. Neste estudo, foi comparada a eficiência de duas metodologias de análises (o enxágue da carcaça e a excisão da pele) para recuperação de células da bactéria, bem como para a verificação da ocorrência do agente patogênico em amostras coletadas na região de Botucatu, SP. Em 100 carcaças analisadas, computando-se os resultados de ambas as metodologias, 43% (43/100) estavam contaminadas por Salmonella spp. Por meio de metodologia de enxágue foram detectadas 81,40% (35/43) amostras com resultados positivos e 51,16% (22/43) pela técnica de excisão. Embora a técnica de enxague tenha detectado número superior de amostras positivas, não houve diferença estatística (p > 0,05) entre os resultados obtidos pelas duas metodologias. Estudos adicionais devem ser realizados, dada à importância das carcaças de frango na veiculação de Salmonella spp. e sua consequente introdução no ambiente de preparação de alimentos