RESUMO
In Latin America, hematophagous bats are the main reservoirs of rabies virus (RABV) to livestock, to other mammals and, occasionally, to human. Nonetheless, reports of exposure of human and pets to RABV upon aggression by non-hematophagous bats are increasing, possibly facilitated by the synanthropic habits of these bats. We, herein, report the detection and genetic identification of a RABV recovered from an insectivorous bat found sick in a student housing building at the Federal University of Santa Maria, Southern Brazil. Taxonomic characterization identified the captured bat as a member of the genus Nyctinomops, family Molossidae, the group of insectivorous bats. Brain fragments of the bat were positive for RABV antigens by fluorescent antibody test (FAT) and for sequences of the nucleoprotein (N) gene by RT-PCR. The N amplicon was submitted to nucleotide sequencing and analysis, showing that the consensus sequences (SV 33/19) had high identity with RABV sequences of insectivorous bats deposited in GenBank. At phylogenetic tree, the N gene sequences of SV 33/19 clustered with RABV recovered from Nyctinomops laticaudatus, Molossus molossus, and Tadarida lauticaudata bats, and a part of RABV variant 3, 4, and 6, that correspond to Desmodus rotundus, Tadarida brasiliensis, and Lasiurus cinereus, respectively. Although no direct human or domestic animal exposure has been reported, this case strengthens the need for a continuous rabies vaccination in pets in the surrounding areas, since non-hematophagous bats may serve as source of infection for these animals. These findings also call attention for continuous monitoring of populations of synanthropic bats to avoid/prevent human exposure.
Assuntos
Quirópteros , Vírus da Raiva , Raiva , Animais , Brasil , Quirópteros/virologia , Filogenia , Raiva/veterinária , Vírus da Raiva/genéticaRESUMO
Candida kefyr has been considered both a food-spoiling agent and a type of yeast with fermentation properties. In this study, the authors have evaluated the antimicrobial activity of a coconut oil-in-water emulsion associated to the presence of C. kefyr. Fresh coconut kernels were used to obtain the coconut oil-in-water emulsion, the sterile coconut oil-in-water emulsion by decantation, and the coconut oil by means of a heating process. Commercial virgin coconut oil was also used. Agar diffusion, minimal inhibitory concentration and minimal bactericidal concentration (MIC/MBC) techniques were employed to evaluate antimicrobial activity against E. coli and S. epidermidis. The C. kefyr isolate was identified and confirmed. Coconut milk-derived fatty acids were characterized by acid index and thin layer chromatography. Scanning electronic microscopy was performed to evaluate the morphology of the microorganisms. Lipase activity of C. kefyr isolate was also detected. Coconut oil-in-water emulsion associated to C. kefyr was active against both bacteria. Thin layer chromatography confirmed the presence of triglycerides and free fatty acids. The acid index showed higher acidity potential for the coconut oil-in-water emulsion. The microscopic images showed antibacterial action through the formation of membrane holes' and demonstrated yeast shape. All the above show new potentials for C. kefyr and coconut oil-in-water emulsion in food technology.
RESUMO
Molecular techniques have revealed a high prevalence of Pneumocystis colonization in wild mammals. Accurate quantification of Pneumocystis sp. is essential for the correct interpretation of many research experiments investigating this organism. The objectives of this study were to detect the presence of Pneumocystis sp. in bats by qPCR, and to distinguish colonization from infection. Probes and primers for real time PCR (qPCR) were designed based on the gene of major surface glycoprotein (MSG) of Pneumocystis sp., in order to analyze 195 lung tissue samples from bats captured (2007-2009). All samples were also analyzed by nested PCR, using oligonucleotide primers designed for the gene encoding the mitochondrial small subunit rRNA (mtSSU rRNA) to confirm the results. The qPCR assay was standardized using a standard curve made with the DNA extracted from bronchoalveolar lavage positive for Pneumocystis jirovecii. The average Ct was found to be between 13 and 14 (calibration curve) for the detection of infection with Pneumocystis sp. and above these values for colonization. It was considered as negative samples the ones that had Ct values equal to 50. Out of the total 195 samples, 47 (24.1%) bat lung DNA samples were positive for Pneumocystis sp. by qPCR. The most common bat species found were: Tadarida brasiliensis (23.4%), Histiotus velatus (17.0%), Desmodus rotundus (14.9%) and Molossus molossus (8.5%). The average cycle threshold of the positive samples (bats) was 25.8 and standard deviation was 1.7. The DNA samples with Ct values greater than 14 suggest that these animals might be colonized by Pneumocystis sp. Results obtained in this study demonstrated the usefulness of the qPCR procedure for identification of Pneumocystis sp. and for distinction between its colonizing or infectious status in bats.
Assuntos
Portador Sadio/veterinária , Quirópteros/microbiologia , Reservatórios de Doenças/microbiologia , Infecções por Pneumocystis/transmissão , Pneumocystis/isolamento & purificação , Animais , Brasil , Líquido da Lavagem Broncoalveolar/microbiologia , Portador Sadio/epidemiologia , Portador Sadio/microbiologia , Quirópteros/classificação , DNA Fúngico/análise , DNA Fúngico/genética , Proteínas Fúngicas/genética , Especificidade de Hospedeiro , Pulmão/microbiologia , Glicoproteínas de Membrana/genética , Pneumocystis/genética , Pneumocystis carinii/genética , Pneumocystis carinii/isolamento & purificação , Reação em Cadeia da Polimerase , Reação em Cadeia da Polimerase em Tempo Real , Ribotipagem , Especificidade da EspécieRESUMO
The biodegradation of effluent from an industrial paper mill was monitored in relation to the parameters total phenols, low molar mass phenols and color during its incubation with laccase (Novozymes), laccase-HBT and peroxidase (Merck). The results revealed that the laccase reduced the effluent color by 37% and the peroxidase by 43%. Also, the enzymes had a great efficiency in the reduction of color, total phenols and high molar mass compounds contained in the effluent, indicating that depolymerization occurred.
A biodegradação dos efluentes de uma indústria de papel e celulose foi monitorada em relação aos parâmetros de fenóis totais, fenóis de baixa massa molar e cor durante a sua incubação com lacase (Novozymes), lacase-HBT e peroxidase (Merck). Os resultados revelaram que a lacase reduziu a cor do efluente em 37% e a peroxidase em 43%. Também foi possível observar que as enzimas possuiram grande eficiência na redução de cor, compostos fenólicos totais e compostos fenólicos de alta massa molar contidos no efluente, indicando, desta forma, que a despolimerização ocorreu.
Assuntos
Biotransformação , Indústria de Papel e Celulose , Efluentes Industriais , Peroxidase/administração & dosagem , Lacase/administração & dosagemRESUMO
ABSTRACT The biodegradation of effluent from an industrial paper mill was monitored in relation to the parameters total phenols, low molar mass phenols and color during its incubation with laccase (Novozymes), laccase-HBT and peroxidase (Merck). The results revealed that the laccase reduced the effluent color by 37% and the peroxidase by 43%. Also, the enzymes had a great efficiency in the reduction of color, total phenols and high molar mass compounds contained in the effluent, indicating that depolymerization occurred.
RESUMO A biodegradação dos efluentes de uma indústria de papel e celulose foi monitorada em relação aos parâmetros de fenóis totais, fenóis de baixa massa molar e cor durante a sua incubação com lacase (Novozymes), lacase-HBT e peroxidase (Merck). Os resultados revelaram que a lacase reduziu a cor do efluente em 37% e a peroxidase em 43%. Também foi possível observar que as enzimas possuiram grande eficiência na redução de cor, compostos fenólicos totais e compostos fenólicos de alta massa molar contidos no efluente, indicando, desta forma, que a despolimerização ocorreu.
RESUMO
A sandwich ELISA (S-ELISA) was developed to detect antibodies to rabies virus in seraof different species. The test was performed as follows: ELISA plates were coated withpolyclonal mouse/anti-rabies antibodies for 2 hours at 37ºC. After adsorption, plateswere washed and non-specific binding blocked with 2% powdered milk. In a separateplate, serial threefold dilutions of test sera were incubated with inactivated rabies virusantigen. The mixtures were then placed on the rabies antibody-coated plates andincubated. These were then washed and incubated with polyclonal rabbit/anti-rabiesantibodies. Subsequently, a rabbit/IgG-peroxidase conjugate was added and platesincubated. After washing, the chromogen (ABTS with 0.15% H2O2) was added to platesand after incubation for 30 min were read in a spectrophotometer (OD405). To validatethe S-ELISA, 128 serum samples including humans, cattle, hematophagous and nonhaematophagous bats, mice, marmosets, ocelots - Leopardus pardalis, raccoons -Procyon lotor, jaguarondi - Herpailurus yaguarondi, fox - Cerdocyon thous and coati -Nasua nasua, were tested and compared to a standard fluorescent antibody virusneutralization test (FAVN). In comparison to FAVN, the S-ELISA showed highsensitivity (82.98%) and specificity (100%), with an accuracy of 87.5%. Subsequently,738 serum samples from different species were tested in the S-ELISA. Antibodies torabies were detected by S-ELISA in all species tested, with the exception of the threeserum samples from raccoons. The S-ELISA was shown to be a serological test of lowcost that can be easily implemented in diagnostic laboratories. In addition, no liveanimals, infectious virus, cell culture or fluorescence microscopy are required forperformance of the test. This is an additional advantage of the S-ELISA over othermethods of rabies antibody detection.