Isolation and characterization of synthetic pyrethroids-degrading bacterial strains from agricultural soil / Isolamento e caracterização de cepas bacterianas degradadoras de piretróides sintéticos de solo agrícola

Pyrethroid pesticides are commonly used for pest control in agriculture setup, veterinary and home garden. They are now posing increased risks to non-targeted organisms associated to human beings due to their considerable use. The present work deals with the isolation of bacteria with tolerance to high concentrations of bifenthrin and cypermethrin from contaminated soil. Enrichment culture technique (bifenthrin concentration = 50-800 mg/L) was used for bacterial isolation. Bacteria that showed growth on minimal media with bifenthrin were also sub-cultured on minimal media with cypermethrin. Bacteria showing luxurious growth on both the pyrethroid, were screened out based on their morphological, biochemical parameters and by API 20NE Kit. Phylogenetic studies revealed that, one bacterial isolate (MG04) belonging to Acinetobacter lwoffii and other five bacterial isolates (MG06, MG05, MG01, MG03 and MG02) cluster with Pseudomonas aeruginosa, Pseudomonas putida respectively. Isolated members of genera Pseudomonas and Acinetobacter could be used for further detailed degradation studies by using FTIR, HPLC-MS or GC-MS analysis.

Os pesticidas piretróides são comumente usados ​​para controle de pragas na agricultura, veterinária e hortas domésticas. Atualmente eles apresentam riscos aumentados para organismos não-alvo associados a seres humanos devido ao seu uso considerável. O presente trabalho analisou o isolamento de bactérias com tolerância a altas concentrações de bifentrina e cipermetrina de solo contaminado. A técnica de cultura de enriquecimento (concentração de bifentrina = 50-800 mg/L) foi utilizada para o isolamento bacteriano. Bactérias que apresentaram crescimento em meio mínimo com bifentrina também foram subcultivadas em meio mínimo com cipermetrina. Bactérias apresentando crescimento luxuoso em ambos os piretróides foram triadas com base em seus parâmetros morfológicos, bioquímicos e pelo Kit API 20NE. Estudos filogenéticos revelaram que, um isolado bacteriano (MG04) pertencente a Acinetobacter lwoffii e outros cinco isolados bacterianos (MG06, MG05, MG01, MG03 e MG02) agrupam-se com Pseudomonas aeruginosa, Pseudomonas putida respectivamente. Membros isolados dos gêneros Pseudomonas e Acinetobacter podem ser usados ​​para estudos de degradação mais detalhados usando análises de FTIR, HPLC-MS ou GC-MS.

In vitro evaluation of probiotic and technological properties of lactic acid bacteria isolated from artisanal cheese produced in the Serra Geral - Minas Gerais - Brazil / Avaliação in vitro de propriedades probióticas e tecnológicas de bactérias ácido-láticas isoladas de queijo artesanal da Serra Geral (MG)

O presente estudo avaliou o potencial probiótico in vitro de bactérias ácido-láticas (BAL) isoladas de queijo artesanal da Serra Geral (MG), com 14 e 21 dias de maturação, leite cru e pingo, utilizados em sua elaboração, e de bancadas de produção. As bactérias foram submetidas aos testes de antibiograma, à tolerância a ácido gástrico artificial e a sais biliares e ao antagonismo contra micro-organismos indicadores. Levilactobacillus brevis (Q521) e Lactococcus lactis (LLSG) apresentaram os melhores resultados e foram selecionados para a produção de leite fermentado. Apenas LLSG foi capaz de fermentar o leite. O produto apresentou qualidade microbiológica e físico-química adequada, exceto para os teores de proteína, segundo a legislação vigente para leites fermentados, demonstrando potencial tecnológico. A partir dos resultados apresentados, sugere-se que testes in vivo sejam realizados para que LLSG possa vir a ser utilizado como probiótico.

Toxoplasma gondii in lactating animals: potential risk to milk consuming population in Khyber Pakhtunkhwa / Toxoplasma gondii em animais lactantes: risco potencial para população consumidora de leite em Khyber Pakhtunkhwa

Toxoplasma gondii is an intracellular zoonotic protozoan parasite usually infects human and animal worldwide. This study aimed to analyze the sero-prevalence of T. gondii in blood of lactating animals and human living in close proximity and also to detect Toxoplasma DNA in unpasteurized milk of the studied animals. A total of 233 blood and milk samples were collected from lactating animals, and 735 blood samples were taken from humans in District Upper Dir, Khyber Pakhtunkhwa, Pakistan. The blood samples were analyzed through ELISA while the milk samples were analyzed by PCR for the presence of T. gondii DNA. A standard questionnaire was introduced to collect the data from the participants. In animals, the reported sero-prevalence was 32.18% for IgM, 17.16% for IgG, and 6.4% for both IgM and IgG. The reported positivity for T. gondii DNA in milk was 14.44%, 34.8%, 20%, and 26% in sheep, goats, cows, and buffaloes, respectively. In the human blood samples, 9.8% were found positive for IgM and 11.2% for IgG while none of the samples was found positive for both IgM and IgG. Overall sero-prevalence reported in females was significantly higher than the male (p< 0.0001) were the significant risk factors associated with T. gondii infections in animals. In conclusion, T. gondii infection is prevalent in lactating animals and humans using their raw milk in the study area. It is suggested that raw milk should be considered as a vehicle for the transmission of T. gondii to humans. Proper pasteurization of milk is very useful in limiting the transmission of infection. Awareness and control programs should be implemented to prevent the infection.

Toxoplasma gondii é um protozoário parasita intracelular zoonótico que geralmente infecta humanos e animais em todo o mundo. Este estudo teve como objetivo analisar a soroprevalência de T. gondii no sangue de animais lactantes e humanos que vivem em proximidade, além de detectar o DNA de Toxoplasma no leite não pasteurizado dos indivíduos estudados. Um total de 233 amostras de sangue e leite foram coletadas de animais lactantes e 735 amostras de sangue foram coletadas de humanos no Distrito Upper Dir Khyber Pakhtunkhwa, no Paquistão. As amostras de sangue foram analisadas pelo método ELISA enquanto as amostras de leite foram analisadas por PCR para a presença de DNA de T. gondii. Um questionário padrão foi introduzido para coletar os dados dos participantes. Em animais, a soroprevalência relatada foi de 32,18% para IgM, 17,16% para IgG e 6,4% para IgM e IgG. A positividade relatada para DNA de T. gondii encontrada no leite foi de 14,44%, 34,8%, 20% e 26% em ovelhas, cabras, vacas e búfalas, respectivamente. Nas amostras de sangue humano, 9,8% foram consideradas positivas para IgM e 11,2% para IgG, enquanto nenhuma das amostras foi considerada positiva para IgM e IgG. A soroprevalência geral relatada em mulheres foi significativamente maior do que em homens (p < 0,05). Neste estudo, contato com gatos (p < 0,0001), hábitos alimentares (p < 0,0001), fonte de água para beber (p < 0,0001) e más condições de higiene (p < 0,0001) foram os fatores de risco significativos associados a infecções por T. gondii em animais. Em conclusão, a infecção por T. gondii é prevalecente em animais lactantes e humanos que utilizam leite cru, isto é, não-pasteurizado, na área de estudo. Sugere-se que o leite não-pasteurizado seja considerado um veículo de transmissão do T. gondii para humanos. A pasteurização adequada do leite é muito útil para limitar a transmissão de infecções. Programas de conscientização e controle devem ser implementados para prevenir a infecção.

Cultivo associado de Leuconostoc mesenteroides lb10. 4 e Lactococcus lactis l4a8: propriedades antimicrobianas e potencial aplicação / Associated culture of Leuconostoc mesenteroides lb10. 4 and Lactococcus lactis l4a8: antimicrobial properties and potential application / Cultivo asociado de Leuconostoc mesenteroides lb10. 4 y Lactococcus lactis l4a8: propiedades antimicrobianas y aplicación potencial

O interesse por produtos lácteos funcionais tem motivado o estudo e a prospecção de novas bactérias lácticas. Neste contexto o objetivo deste trabalho foi explorar a potencial associação de Leuconostoc mesenteroides LB10.4 e Lactococcus lactis L4A8 isolados de leite de búfala e identificar aplicações em matriz alimentar. Foram realizados testes de atividade antimicrobiana, influência das bacteriocinas e avaliação da eficiência das bactérias ácido lácticas (BAL) individualmente e associadas frente a espécie de Listeria monocytogenes ATCC 7644 aplicadas em caldo Tryptic Soy Broth e em matriz láctea. Na avaliação da atividade antimicrobiana, Leuconostoc mesenteroides LB 10.4 e Lactococcus lactis L4A8 foram capazes de inibir Staphylococcus aureus ATCC 25923 e Listeria monocytogenes ATCC 7644 com halos de inibição variando de 8 a 16 mm e 6 a 18 mm, respectivamente, pelos dois métodos testados. Na avaliação do efeito das bacteriocinas, os resultados demonstraram melhor controle inibitório dos patógenos pela nisina nas concentrações de 1% e 2%, com halos de inibição entre 14 a 24 mm. A avaliação da eficiência das BAL individualmente e associadas frente a espécie de Listeria monocytogenes ATCC 7644, demonstrou que os isolados em associação são capazes de inibir com mais efetividade a bactéria patogênica, sendo observada uma redução na contagem de L. monocytogenes de 2,67x107 UFC/g para 1,35x104 UFC/g após 240 horas, em matriz alimentar. As bactérias lácticas Leuconostoc mesenteroides LB10.4 e Lactococcus lactis L4A8 apresentaram características promissoras quanto ao seu potencial inibitório, não sendo inibidas pela pediocina. Destaca-se com esses resultados, a importância de estudar o leite de búfala como fonte de novos candidatos de bactérias lácticas autóctones para aplicação em alimentos.(AU)

The interest in functional dairy products has motivated the study and prospection of new lactic bacteria. In this context, the aim of this study was to explore the potential association of Leuconostoc mesenteroides LB10.4 and Lactococcus lactis L4A8 isolated from buffalo milk and to identify applications in food matrix. Tests of antimicrobial activity, influence of bacteriocins and evaluation of the efficiency of lactic acid bacteria (LAB) individually and associated against the species of Listeria monocytogenes ATCC 7644 applied in Tryptic Soy Broth and in milk matrix were performed. In the evaluation of antimicrobial activity, Leuconostoc mesenteroides LB 10.4 and Lactococcus lactis L4A8 were able to inhibit Staphylococcus aureus ATCC 25923 and Listeria monocytogenes ATCC 7644 with inhibition halos ranging from 8 to 16 mm and 6 to 18 mm, respectively, by the two methods tested. In evaluating the effect of bacteriocins, the results showed better inhibitory control of pathogens by nisin at concentrations of 1% and 2%, with inhibition halos between 14 and 24 mm. The evaluation of the efficiency of LAB individually and associated against the species of Listeria monocytogenes ATCC 7644, showed that the isolates in association are able to more effectively inhibit the pathogenic bacteria, with a reduction in the L. monocytogenes count of 2.67x107 CFU /g to 1.35x104 CFU/g after 240 hours, in food matrix. The lactic acid bacteria Leuconostoc mesenteroides LB10.4 and Lactococcus lactis L4A8 showed promising characteristics regarding their inhibitory potential, not being inhibited by pediocin. With these results, attract attention the importance of studying buffalo milk as a source of new candid ates of autochthonous lactic acid bacteria for application in food.(AU)

El interés por los productos lácteos funcionales ha motivado el estudio y prospección de nuevas bacterias ácido lácticas. En este contexto, el objetivo de este trabajo fue explorar la asociación potencial de Leuconostoc mesenteroides LB10.4 y Lactococcus lactis L4A8 aislados de leche de búfala e identificar aplicaciones en matriz alimentaria. Se realizaron pruebas de actividad antimicrobiana, influencia de bacteriocinas y evaluación de la eficiencia de bacterias ácido lácticas (BAL) individualmente y asociadas frente a las especies de Listeria monocytogenes ATCC 7644 aplicadas en caldo Tryptic Soy Broth y en matriz de leche. En la evaluación de la actividad antimicrobiana, Leuconostoc mesenteroides LB10.4 y Lactococcus lactis L4A8 fueron capaces de inhibir Staphylococcus aureus ATCC 25923 y Listeria monocytogenes ATCC 7644 con halos de inhibición de 8 a 16 mm y de 6 a 18 mm, respectivamente, por los dos métodos probados. Al evaluar el efecto de las bacteriocinas, los resultados mostraron un mejor control inhibitorio de patógenos por parte de la nisina a concentraciones de 1% y 2%, con halos de inhibición entre 14 y 24 mm. La evaluación de la eficiencia de las BAL de forma individual y asociada frente a la especie de Listeria monocytogenes ATCC 7644, mostró que los aislados en asociación son capaces de inhibir más eficazmente las bacterias patógenas, con una reducción del recuento de L. monocytogenes de 2,67x107 UFC/g a 1,35x104 UFC/g después de 240 horas, en matriz alimentaria. Las bacterias del ácido láctico Leuconostoc mesenteroides LB10.4 y Lactococcus lactis L4A8 mostraron características promisorias en cuanto a su potencial inhibidor, no siendo inhibidas por la pediocina. Con estos resultados, se destaca la importancia de estudiar la leche de búfala como fuente de nuevos candidatos de bacterias ácido lácticas autóctonas para su aplicación en alimentos.(AU)

Microbiological featuring of two neotropical fish species from the quilombola area of maranhão state, Brazil / Caracterização microbiológica de duas espécies de peixes neotropicais oriundas de área quilombola maranhense, Brasil

The aim of study is to feature the microbiological quality of two neotropical fish species from the quilombola area of Maranhão State, Brazil. In order to do so, 21 samples of Hoplerythrinus unitaeniatus and 21 samples of Cichlasoma bimaculatum were captured in flooded environment. Collected fish were euthanized in laboratory environment; muscle fragments were removed for microbiological analyses focused on enumerating molds and yeasts, viable strict and facultative mesophilic microorganisms and coagulase-positive staphylococci; on counting total and thermotolerant coliforms; and on investigating Escherichia coli and Salmonella sp. Microbiological results were compared to the Brazilian legislation, which establishes the list of microbiological standards for food products. Among the assessed fish, 9.52% were classified as non-acceptable for human consumption, based on the Salmonella parameter. Enumerated coagulase-positive staphylococci ranged from < 10 to 3.9 x 104 CFU/g; 9.52% of assessed fish were classified as having intermediate standard for human consumption, whereas 4.76% were classified as non-acceptable for such a purpose. E. coli counting ranged from 3.6 to > 1,100MPN/g; 4.76% of assessed fish were classified as having intermediate standard for human consumption, whereas 4.76% were classified as non-acceptable for such a purpose. Total and thermotolerant coliforms' counting and the enumeration of viable strict and facultative aerobic microorganisms, as well as of molds and yeasts, have evidenced high microbial population rates; this finding suggests poor hygienic conditions at capture site, contaminated raw material and risk of incidence of enteropathogens. This finding has evidenced imbalance in the investigated environment, as well as compromised aquatic biodiversity.(AU)

Objetivou-se caracterizar a qualidade microbiológica de duas espécies de peixes neotropicais oriundas de área qui-lombola maranhense, Brasil. Para isso, foram capturadas 21 amostras de Hoplerythrinus unitaeniatus e 21 amostras de Cichlasoma bimaculatum de ambiente alagável. No laboratório, os peixes foram eutanasiados, procedida a retirada dos fragmentos mus-culares e realizada as análises microbiológicas: enumeração de bolores e leveduras, micro-organismos mesófilos aeróbios estri-tos e facultativos viáveis e de estafilococos coagulase positiva, quantificação de coliformes totais e termotolerantes, pesquisa de Escherichia coli e Salmonella sp. Os resultados microbiológicos obtidos foram comparados com a legislação brasileira que estabelece a lista de padrões microbiológicos para alimentos. Dos peixes avaliados, 9,52% foram considerados inaceitáveis para consumo humano para o parâmetro Salmonella. A enumeração de estafilococos coagulase positivavariou de <10 a 3,9 x 104 UFC/g, sendo 9,52% dos peixes considerados com padrão intermediário e 4,76% inaceitáveis para consumo. A quanti-ficação de E. coli variou de 3,6 a >1.100 NMP/g, sendo 4,76% considerados com padrão intermediário e 4,76% inaceitáveis para consumo. A quantificação de coliformes totais e termotolerantes e a enumeração de micro-organismos aeróbios estritos e facultativos viáveis e bolores e leveduras revelou elevadas populações microbianas o que sugere más condições higiênicas do local de captura, matéria-prima contaminada e risco da presença de enteropatógenos, demonstrando desequilíbrio no ambiente estudado com comprometimento da biodiversidade aquática.(AU)

The Effect of Moringa Oleifera Leaf and Canola Seed Powder on Fattening, Laying Performance, Blood Plasma Constituents, and Microbiota in Japanese Quails

The aim of the study was to investigate the effect of feeding Japanese quail chicks with diets containing different levels of Moringa oleifera leaf and canola seed powder on growth performance, carcass yield, blood plasma constituents, and egg production. The trial lasted for a total of 7 weeks, with 5 weeks of fattening and two weeks of laying. The first group was the control group, while the second group was supplemented with Moringa oleifera leaves (4g/kg diet), the third group was supplemented with canola seed powder (4g/kg diet), the fourth group was supplemented with a mix of Moringa oleifera leaves and canola seed powder (8g/kg diet). The results showed that canola seed powder from 0-3 weeks of age increased body weight in comparison to the control group, but there were no significant differences (p<0.05) among groups in terms of the final body weight and feed conversion ratio. Average daily feed intake was significantly different (p<0.05). However, body weight, carcass weight, liver weight, gizzard weight, and abdominal fat weight increased significantly compared to the control group. Feeding Moringa oleifera leaves and canola seed powder significantly increased the total plasma protein, as compared to the control group. There were significant decreases in cholesterol, triglycerides and HDL levels among groups, with no significant differences in glucose, ALT and LDL among all treatments. The addition of Moringa oleifera leaf and canola seed powder showed significant effects on calcium and magnesium. Both Moringa oleifera leaf and Moringa-canola mixture significantly reduced the presence of some pathogenic bacteria in the digestive system, which was seen as an important contribution to the digestive and immune systems.(AU)

Infectious diseases of neotropical primates

Neotropical primates are represented by more than 200 species and subspecies distributed in five families. Considering that some of these species are considered endangered, disease investigation in these populations is critical for conservation strategies. Therefore, an increasing number of studies and publications on this topic became available in the past few years. This review deals with infectious diseases of neotropical primates, with focus on free-ranging animals, including those caused by bacterial, viral, protozoal, metazoan, or mycotic infectious organisms, with particular emphasis on gross and microscopic lesions associated with these diseases. In addition, a few relevant unpublished cases of infection by Staphylococcus spp., Streptococcus spp., E. coli and Pseudomonas spp. were included in this review.(AU)

Fatal and disseminated infection by Entamoeba invadens in a red-footed tortoise (Chelonoidis carbonaria)

Amebiasis is an important parasitosis that can affect reptiles, specially caused by protozoas of the genus Entamoeba, which include pathogenic or commensal species. Entamoeba invadens is the most common amoeba to cause serious disease and death in reptiles. This paper aims to report a case of a sudden death due to a disseminated infection by Entamoeba invadens in a red-footed tortoise (Chelonoidis carbonaria). The animal was brought to the Center for Management and Conservation of Wild Animals of the Fauna Division and found dead after being kept in an enclousure with other captive tortoises for 11 months. Macroscopic findings evidenced necrotizing typhlitis and proctitis and round yellow areas in the right lobe of liver parenchyma. In the histological examination, necrotizing and heterophilic enteritis and necrotizing hepatitis with macrovesicular degeneration of hepatocytes, associated with mixed inflammatory infiltrate were present. Both organs revealed numerous amoebic trophozoites, morphologically suggestive of the genus Entamoeba, and bacterial colonies. The agent was confirmed by PCR and Sanger DNA sequencing, which leads this study to be the first confirmed case report of E. invadens infection in Brazil in a red-footed tortoise.(AU)

Genetic variability of Mycoplasma synoviae detected in commercial layers in southeastern Brazil / Variabilidade genética de Mycoplasma synoviae detectado em poedeiras comerciais do sudeste do Brasil

Este estudo teve como objetivo caracterizar genótipos de M. synoviae circulantes em poedeiras comerciais, na Região Sudeste do Brasil. Para estabelecer a relação evolutiva entre as cepas de MS, foi analisada a sequência genética do gene vlhA de 11 cepas de MS identificadas no Rio de Janeiro e em São Paulo, de 10 outras cepas de MS identificadas no Brasil e cujas sequências foram depositadas no banco de dados GenBank, e da cepa vacinal MSH. O método da máxima verossimilhança e o modelo de Kimura com dois parâmetros foram utilizados para comparar as cepas. As sequências obtidas foram depositadas no Genbank, sob os números de acesso OP279775 a OP279785. Foi possível verificar a presença de diferentes cepas circulantes no Brasil, com alta similaridade entre as cepas do Rio de Janeiro pela análise do gene vlhA. As duas cepas paulistas detectadas no presente estudo possuem o baixo percentual (68%) de similaridade, demonstrando a variabilidade das cepas dessa localidade.

Quality of refrigerated raw milk according to the bulk transport conditions

Technical regulations that determine milk quality standards in Brazil establish the minimum criteria that the processing industry's raw material must present. These rules can challenge the logistics sector (transport) to maintain adequate milk conservation levels. We aimed to evaluate the influence of bulk transport and storage conditions on refrigerated raw milk quality. The experiment was carried out in a dairy industry in Santa Catarina State. Information and samples of refrigerated raw milk were collected from dairy farms, from isothermal compartments of milk trucks, and from the industry's storage silo. These samples were submitted to microbiological analysis, somatic cell count(SCC), and physicochemical analysis. The results were evaluated by ANOVA and multivariate analysis (factor analysis). The transport of raw milk in bulk and the transfer of raw material to the industrial silo worsened the milk's microbiological quality (p < 0.0001) for standard plate count (SPC) and psychrotrophic microorganism count(PMC), without affecting, the physicochemical characteristics of the milk (p > 0.05). Poor hygienic conditions in milk storage rooms, in the refrigeration bulk tanks of the dairy farms, and during the transport, as well as the transport on longer routes, were related to higher SPC and psychrotrophic count while receiving milk by the dairy industry with higher temperatures was only associated with higher SPC.(AU)

Short communication: psychrotrophic microorganism count in raw milk samples preserved with azidiol

The study aimed to evaluate the use of the azidiol® preservative for psychrotrophic microorganism count (PMC) in cooled raw milk. Two studies were carried out, one under controlled conditions (experiment 1) and the other under field conditions (experiment 2), in which samples of raw milk were taken with and without the use of the azidiol® preservative and analyzed at predefined times (0, 6, 12 and 24 hours -experiment 1) and at varying times (experiment 2). Analysis of variance and regression analysis using SAS were applied for data statistical analysis. Milk samples without azidiol®showed higher PMC with increasing time between sampling and analysis, while in samples preserved with azidiol®, this count remained constant. Samples of cooled raw milk intended for PMC should be collected in flasks containing the azidiol®preservative.(AU)

Salmonella Serovars Isolated from Poultry Breeding Flocks under the Brazilian Official Control Programme Between 2016 and 2018

The control of Salmonella in the poultry production chain combined with biosecurity measures is an important tool to maintain and guarantee the sanitary status of Brazilian flocks. The aim of this work was to compare official laboratory data on molecular typification of Salmonella isolates from poultry breeding flocks in different Brazilian states between 2016 and 2018 and identify the production category with the most positive flocks, in light of current legislation. Surveillance data of positive samples from the official Brazilian Salmonella Control Programme sent to Federal Agricultural Defence Laboratory of São Paulo (LFDA-SP) after molecular characterization were analysed. These data were subject to an exploratory study, undergoing a descriptive statistical analysis followed by the use of frequency and non-parametric hypothesis tests. Overall, 49 serovars were detected in poultry broiler-breeder and layer-breeder flocks. Salmonella ser. Heidelberg, Salmonella ser. Anatum, Salmonella ser. Newport, Salmonella ser. Schwarzengrund and Salmonella ser. Mbandaka were the five most common isolated serovars. The data shows that there is an opportunity to improve biosecurity measures in parent breeder flocks. A total of 16 serovars were identified in turkey-breeders. Salmonella ser. Anatum, Salmonella ser. Newport, Salmonella ser. Brandenburg, Salmonella ser. Litchfield, and Salmonella ser. Livingstone were the most common ones. The four official controlled serovars represented a small part of the isolated strains. These data demonstrate the importance of an official program in Brazil for Salmonella surveillance in breeder flocks combined with biosecurity measures.(AU)

Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum

Starch processing industries use amylases, accounting for approximately 30% of the world's enzyme market. Previously, an amylase-producing strain of Epicoccum nigrumwas isolated from maize grains. Although E. nigrumamylase production is already reported in the literature, no published data on production optimization or characterization of the produced enzyme exists. The objectives of this work were to improve the amylase production by theE. nigrumPG 16 strain and to purify and characterize the produced enzyme. The E. nigrumPG 16 amylase production best conditions in submerged culture were: inoculum of 4% (v v-1) of a five-days-old stationary culture homogenate, agitation at 100 rpm, 25°C, natural light, 72 hours of incubation, starch as thecarbon source, and an initial medium pH of 7.0. A molecular exclusion chromatography profile has shown the production of only one amylase, which was partially purified with ammonium precipitation and dialysis. The enzyme optima pH and temperature are 6.0 and 50°C, respectively. The partially purified enzyme lostits activity when incubated for 30 min in temperatures above 40°C, presenting a T50of 46.25°C. The KMand Vmaxof the partially purified enzyme are 1.72 mg mL-1of starch and 0.15 mgmin-1of degraded starch, respectively. The ion Ca2+slightly activated the studied enzyme. The ions Cu2+, Zn2+, and Fe3+and the detergents SDS and Tween 80 acted as inhibitors of the studied enzyme. The partially purified enzyme released glucose from p-nitrophenyl α-D-glucopyranoside (p-NPG). Glucose was the enzyme's main product from starch hydrolysis, as evidenced by thin-layer chromatography. The E. nigrumPG 16 studied enzyme is a glucoamylase and represents an alternative for enzymatic starch hydrolysis.(AU)

Occurrence of Apicomplexa protozoa in wild birds in the Northeast region of Brazil / Ocorrência de protozoários Apicomplexa em aves silvestres na região Nordeste do Brasil

Protozoa of the Apicomplexa phylum are worldwide distributed with capacity to infect endothermic animals. The study of these protozoa in wild birds in Brazil is scarce. This study aimed to evaluate the occurrence of apicomplexan protozoa in wild birds in the Northeast of Brazil. From October to December 2019, brain tissue samples were collected from 71 captive birds from the Wild Animal Screening Center of the Pernambuco State (CETRAS-Tangara) and 25 free-living birds from the Caatinga biome in Rio Grande do Norte, totaling 96 animals (41 species). Brain fragments were subjected to molecular diagnosis by nested PCR for the 18s rDNA gene of Apicomplexa parasites, followed by DNA sequencing. This gene was detected in 25% (24/96) of the samples, and it was possible to perform DNA sequencing of 14 samples, confirming three genera: Isospora, Sarcocystis and Toxoplasma from eight bird species (Amazona aestiva, Coereba flaveola, Egretta thula, Paroaria dominicana, Sporophila nigricollis, Cariama cristata, Columbina talpacoti, Crypturellus parvirostris). The occurrence these coccidia in wild birds provides important epidemiological information for the adoption of preventive measures for its conservation. Future studies are needed to better understand the consequence of Apicomplexa infection in birds in Caatinga and Atlantic Forest biomes.(AU)

Protozoários do filo Apicomplexa são distribuídos mundialmente e com capacidade de infectar animais endotérmicos. O estudo destes protozoários, em aves silvestres do Brasil, é escasso. Objetivou-se avaliar a ocorrência de protozoários Apicomplexa em aves silvestres na região Nordeste do Brasil. De outubro a dezembro de 2019, foram coletadas amostras de encéfalo de 71 aves de cativeiro do Centro de Triagem e Reabilitação de Animais Silvestres de Pernambuco (CETRAS-Tangara). E 25 aves de vida livre do bioma Caatinga no Rio Grande do Norte, totalizando 96 animais (41 espécies). Os fragmentos de encéfalo foram submetidos ao diagnóstico molecular por nested PCR, para o gene 18s rDNA de protozoários Apicomplexa, seguido por sequenciamento do DNA. Este gene foi detectado em 25% (24/96) das amostras analisadas; foi possível realizar o sequenciamento de 14 amostras, confirmando-se três gêneros: Isospora, Sarcocystis e Toxoplasma em oito espécies de aves (Amazona aestiva, Coereba flaveola, Egretta thula, Paroaria dominicana, Sporophila nigricollis, Cariama cristata, Columbina talpacoti, Crypturellus parvirostris). A ocorrência destes coccídios nas aves silvestres fornece informações epidemiológicas importantes para a adoção de medidas preventivas tendo em vista sua conservação. Estudos futuros são necessários para melhor compreensão da consequência da infecção por Apicomplexa, em aves silvestres dos biomas Caatinga e Floresta Atlântica.(AU)

Monitoring periodontal lesions and their effects during pregnancy: microbiological aspects of the oral cavity and amniotic fluid in pregnant ewes / Monitoramento de lesões periodontais e seus efeitos na gestação: aspectos microbiológicos da cavidade bucal e líquido amniótico de ovelhas gestantes

Periodontitis affects the teeth supporting tissues, leading to tooth loss and damage to animal health. Evidence in humans suggests that oral microorganisms spread systemically, increasing the risk of pregnancy disorders such as miscarriage, prematurity, and low birth weight. This study aimed to verify whether periodontopathogenic microorganisms reach the transplacental unit, culminating in problems in pregnant ewes. After analyzing the oral cavity, 10 clinically healthy pregnant ewes (OGCH group) and 10 pregnant ewes with periodontitis (OGP group) were selected. The subgingival biofilm was collected for the polymerase chain reaction (PCR) test and amniotic fluid for both the PCR and interleukin (IL) analysis. Peripheral blood was collected for complete blood count, and analyses of IL-6, IL1-ß, and tumor necrosis factor-α were performed. Placental fragments were collected to assess the inflammatory changes using optical microscopy. After giving birth, both the ewes and their lambs were weighed. On clinical examination, a positive correlation between bleeding and suppuration (correlation index - CI=0.54), suppuration and marginal gingivitis (CI=0.34), and marginal gingivitis and edema (CI=0.54) was observed. The weights of the ewes (p=0.013) and their respective lambs (p=0.04) in the OGP group were lower than those of their OGCH group counterparts. The hematological analysis revealed that the OGP group ewes showed a slight increase in the mean corpuscular volume (p=0.2447), segmented cells (p=0.3375), and eosinophils (p=0.3823) when compared with the OGCH group ewes, without a statistical difference. Regarding the microorganisms detected in the oral cavity, there was a significant difference between the occurrence of periodontal pockets and the presence of Fusobacterium necrophorum (p=0.0328), Porphyromonas asaccharolytica (p=0.0392), and the Mollicutes class (p=0.0352). Staphylococcus genus (p=0.9107) and Archaea domain (p=0.7245) were detected in the amniotic samples of both groups, without a significant difference, whereas P. asaccharolytica (p=0.2685) was only detected in one sample in the OGCH group. The expression of cytokine IL-6 in the OGP group differed significantly between the prepartum and postpartum periods (p=0.0039); moreover, it differed significantly in the postpartum period between the OGCH and OGP groups (p=0.0198). Histological examination showed a higher percentage of placental changes in the OGP group (70%) than in the OGCH group, such as the presence of macrophages, neutrophils, plasma cells, and multifocal areas of calcification. These results do not corroborate the hypothesis of dissemination of oral microorganisms to the placental unit, suggesting that it constitutes placental isolation in sheep.

A periodontite afeta os tecidos de suporte dos dentes levando à perda dentária e danos à saúde do animal. Evidências em humanos sugerem que os microrganismos orais se espalham sistemicamente, aumentando o risco de distúrbios da gravidez, como aborto espontâneo, prematuridade e baixo peso ao nascer. Este estudo teve como objetivo verificar se microrganismos periodontopatogênicos atingem a unidade transplacentária, culminando em problemas em ovelhas gestantes. Após análise da cavidade oral, foram selecionadas 10 ovelhas gestantes clinicamente saudáveis (grupo OGCH) e 10 ovelhas gestantes com periodontite (grupo OGP). O biofilme subgengival foi coletado para o teste de reação em cadeia da polimerase (PCR) e o líquido amniótico para teste de PCR e análise de interleucina (IL). Sangue periférico foi coletado para hemograma completo e análises de IL-6, IL1-ß e fator de necrose tumoral-α foram realizadas. Fragmentos de placenta foram coletados para avaliação das alterações inflamatórias por meio de microscopia óptica. Após o parto, as ovelhas e seus cordeiros foram pesados. Ao exame clínico, observou-se correlação positiva entre sangramento e supuração (índice de correlação - IC=0,54), supuração e gengivite marginal (IC=0,34) e gengivite marginal e edema (IC=0,54). Os pesos das ovelhas (p=0,013) e de seus respectivos cordeiros (p=0,04) do grupo OGP foram inferiores aos do grupo OGCH. A análise hematológica revelou que as ovelhas do grupo OGP apresentaram discreto aumento no volume corpuscular médio (p=0,2447), células segmentadas (p=0,3375) e eosinófilos (p=0,3823) quando comparadas com as ovelhas do grupo OGCH, sem diferença estatística diferença. Em relação aos microrganismos detectados na cavidade oral, houve diferença significativa entre a ocorrência de bolsas periodontais e a presença de Fusobacterium necrophorum (p=0,0328), Porphyromonas asaccharolytica (p=0,0392) e da classe Mollicutes (p=0,0352). O gênero Staphylococcus (p=0,9107) e o domínio Archaea (p=0,7245) foram detectados nas amostras amnióticas de ambos os grupos, sem diferença significativa, enquanto P. asaccharolytica (p=0,2685) foi detectado apenas em uma amostra do grupo OGCH. A expressão da citocina IL-6 no grupo OGP diferiu significativamente entre os períodos pré e pós-parto (p=0,0039); além disso, diferiu significativamente no período pós-parto entre os grupos OGCH e OGP (p=0,0198). O exame histológico mostrou maior porcentagem de alterações placentárias no grupo OGP (70%) do que no grupo OGCH, como a presença de macrófagos, neutrófilos, plasmócitos e áreas multifocais de calcificação. Esses resultados não corroboram a hipótese de disseminação de microrganismos orais para a unidade placentária, sugerindo que se trata de um isolamento placentário em ovinos.

High Genetic Similarity Among Salmonella Heidelberg Isolated from Poultry Farms, Wild Animals, Beef, Poultry and Pork Meat, and Humans in Brazil

Salmonellosis is an important gastrointestinal infection in humans and cause of foodborne outbreaks in the world. In this context, molecular characterization is essential to understand how the strains circulate. The aim of this study was to evaluate the genotypic distribution of S. Heidelberg according to the source of isolation. The genetic relatedness of the S. Heidelberg isolates was determined by pulsed-field gel electrophoresis (PFGE). The most prevalent pulsotypes of cluster A were BRJF6X01.006 (27/95 = 28,42%) related between 1995 and 2011 in broilers, poultry meat and poultry farms, meat product and human, and BRJF6X01.001 (21/95 = 22,10%) related between 2011 and 2017 in wild animals, broilers, poultry meat, poultry farms, meat product, animal feed, and pork meat. The pulsotype BRJF6X01.001 shows a high distribution in the environmental and productive chain. The degree of similarity between pulsotypes BRJF6X01.006 and BRJF6X01.001 is 88%. To ensure the safety of human and animal health, holistic approaches, including surveillance of Salmonella throughout the environment and in the production chain, together with control measures, are critical. As transmission of Salmonella from food producing animals to wildlife and to the environment is considered potential public health problem, information on the survival and persistence of Salmonella in the environment and in potential reservoirs is of considerable importance.(AU)

Isolation and screening of chromium resistant bacteria from industrial waste for bioremediation purposes / Isolamento e triagem de bactérias resistentes ao cromo de resíduos industriais para fins de biorremediação

Abstract Chromium (VI) a highly toxic metal, a major constituent of industrial waste. It is continuously release in soil and water, causes environmental and health related issues, which is increasing public concern in developing countries like Pakistan. The basic aim of this study was isolation and screening of chromium resistant bacteria from industrial waste collected from Korangi and Lyari, Karachi (24˚52ʹ46.0ʺN 66˚59ʹ25.7ʺE and 24˚48ʹ37.5ʺN 67˚06ʹ52.6ʺE). Among total of 53 isolated strains, seven bacterial strains were selected through selective enrichment and identified on the basis of morphological and biochemical characteristics. These strains were designated as S11, S13, S17, S18, S30, S35 and S48, resistance was determined against varying concentrations of chromium (100-1500 mg/l). Two bacterial strains S35 and S48 showed maximum resistance to chromium (1600 mg/l). Bacterial strains S35 and S48 were identified through 16S rRNA sequence and showed 99% similarity to Bacillus paranthracis and Bacillus paramycoides. Furthermore, growth condition including temperature and pH were optimized for both bacterial strains, showed maximum growth at temperature 30ºC and at optimum pH 7.5 and 6.5 respectively. It is concluded that indigenous bacterial strains isolated from metal contaminated industrial effluent use their innate ability to transform toxic heavy metals to less or nontoxic form and can offer an effective tool for monitoring heavy metal contamination in the environment.

Resumo O cromo (VI), metal altamente tóxico, é um dos principais constituintes dos resíduos industriais. É liberado no solo e na água, causa problemas ambientais e de saúde de crescente preocupação pública em países em desenvolvimento como o Paquistão. O objetivo básico deste estudo foi o isolamento e a triagem de bactérias resistentes ao cromo de resíduos industriais coletados em Korangi e Lyari, Karachi (24˚52'46,0"N 66˚59'25,7"E e 24˚48'37,5"N 67˚06'52,6"E). Do total de 53 cepas isoladas, sete cepas bacterianas foram selecionadas por enriquecimento seletivo e identificadas com base em características morfológicas e bioquímicas. Essas cepas foram designadas como S11, S13, S17, S18, S30, S35 e S48, apresentaram alta resistência aos metais contra concentrações variáveis (100-1500 mg / l) de cromo. Já as cepas S35 e S48 foram identificadas por meio da sequência 16S rRNA e apresentaram 99% de similaridade com Bacillus paranthracis e Bacillus paramycoides. Além disso, as condições de crescimento incluindo temperatura e pH foram otimizadas e ambas as cepas bacterianas apresentaram crescimento máximo na temperatura de 30 ºC, enquanto seu pH ótimo foi observado em 7,5 e 6,5, respectivamente. Conclui-se que o potencial de resistência dessas bactérias resistentes ao cromo pode ser efetivamente utilizado na remoção de cromo de efluentes industriais contaminados. Técnicas de base biológica usando bactérias ajudarão a fornecer métodos mais baratos e ecológicos de remoção, recuperação e desintoxicação de cromo.

Antagonistic activity of lactic acid bacteria isolated from Minas artisanal cheeses against Brucella abortus / Atividade antagonista de bactérias lácticas isoladas de queijos Minas artesanais contra Brucella abortus

This study aimed to evaluate methods for studying the in vitro antimicrobial activity of lactic acid bacteria (LAB) against Brucella abortus and to evaluate the antagonistic effect of LAB on the viability of this pathogen. A total of 18 LAB strains (Lactobacillus plantarum, n = 11; Pediococcus acidilactici, n = 1; Lactobacillus rhamnosus, n = 4; and Lactobacillus brevis,n = 2), isolated from Minas artisanal cheeses produced in three regions (Canastra, Campos das Vertentes, and Araxá) of Minas Gerais State, Brazil, were tested for their antimicrobial activity against B. abortus using three methods: spot-on-lawn, agar well diffusion assay, and antagonistic activity of the culture supernatants. None of the tested LAB strains could inhibit B. abortus in the spot-on-lawn and agar-well diffusion assays. The supernatants produced by LAB had an acidic pH, with intensity depending on bacterial growth and strain, and could inhibit the growth of B. abortus. In contrast, pH-neutralized (pH 7.0) LAB supernatants did not suppress the growth of B. abortus. The results showed that the best technique to study the in vitro antagonism of LAB against B. abortus was the antagonistic activity of culture supernatants. The growth of B. abortus may have been inhibited by acid production.(AU)

Este estudo teve como objetivo avaliar métodos de estudo in vitro da atividade antimicrobiana de bactérias lácticas contra Brucella abortus e avaliar o efeito antagônico das mesmas sobre a viabilidade deste patógeno. Um total de 18 amostras de bactérias lácteas (Lactobacillus plantarum, n = 11; Pediococcus acidilactici, n = 1; Lactobacillus rhamnosus, n = 4; e Lactobacillus brevis, n = 2), isoladas de exemplares de Queijo Minas Artesanal produzidos em três regiões (Canastra, Campos das Vertentes e Araxá) do estado de Minas Gerais, Brasil, foram testados quanto à sua atividade antimicrobiana contra B. abortus usando três métodos: spot-on-lawn, ensaio de difusão em poço e atividade antagonista de sobrenadante de cultura. Nenhuma das cepas testadas foi capaz de inibir B. abortus nos ensaios spot-on-lawm e de difusão em poço. Os sobrenadantes produzidos pelas bactérias lácteas apresentaram pH ácido, com intensidade dependente do crescimento bacteriano e da amostra, podendo inibir o crescimento de B. abortus. Em contraste, os sobrenadantes com pH neutralizado (pH 7,0) não inibiram o crescimento de B. abortus. Os resultados mostraram que a melhor técnica para estudar o antagonismo in vitro de bactérias lácteas contra B. abortus foi a atividade antagonista de sobrenadante de cultura. O crescimento de B. abortus pode ter sido inibido pela produção de ácido.(AU)

Resistance profile of bacterial isolates from different llama microbiotas (Lama glama Linnaeus 1758) / Perfil de resistência de isolados bacterianos de diferentes microbiotas de lhamas (Lama glama Linnaeus 1758)

Este trabalho objetivou o isolamento e a identificação das bactérias de diferentes microbiotas de lhamas. Para tanto, testes de disco difusão e diagnósticos moleculares para pesquisa de genes de resistência foram realizados. Foram isolados cinco Staphylococcus spp. coagulase positiva e 19 Staphylococcus spp. coagulase negativa, 19 Escherichia coli¸ duas Pantoea agglomerans, uma Koserella trabulsii, uma Enterobacter aerogenes e uma Klebsiella Pneumoniae. Entre os isolados de Staphylococcus spp., 79,17% foram resistentes ao sulfazotrim, 45,83% resistentes à penicilina e 20,83% à ampicilina. Foi confirmada a presença do gene mecA em apenas um isolado oxacilina resistente. No teste de disco difusão, 58,3% das enterobactérias foram resistentes à amoxicilina + ácido clavulânico e à cefotaxima, 50% à ceftazidima e ceftriaxona, e 33,3% à amoxicilina. Ainda entre os isolados Gram-negativos, não houve a expressão fenotípica de isolados ESBL. O presente trabalho expôs a presença de microrganismos resistentes a antibióticos em lhamas que não tiveram contato prévio com essas drogas, além da presença do gene mecA em um dos animais. O conhecimento da microbiota bacteriana de diferentes espécies animais tem se tornado cada vez mais importante. Tal relevância se deve à possibilidade de esses microrganismos serem compartilhados entre os animais, os humanos e até mesmo o meio ambiente.

Cardiac hypertrophy associated with Corynebacterium spp. in Didelphis albiventris cubs

The purpose of this manuscript is to describe the clinical-pathological aspects of cardiac hypertrophy related to the presence of Corynebacterium spp. in three Didelphis albiventris cubs. In necropsy, macroscopically, in the heart, cardiomegaly, concentric hypertrophy of the ventricles and the ventricular septum were observed, with consequent reduction of the chamber. Microscopically, the primary lesions found in the heart were cardiomyocyte hypertrophy and necrosis, myocytolysis, and the presence of myriad basophilic bacteria. Liver fragments and endocardial swabs were sent for bacterial culture, in which pleomorphic Gram-positive rods grew, forming small and hemolytic colonies. Chemical tests demonstrated characteristics compatible with Corynebacterium spp. Thus, this report represents the first description of cardiac hypertrophy associated with Corynebacterium spp. in white-eared opossums cubs, representing an essential contribution to studying diseases in wild animals.(AU)