Resumo

Tomatoes (Solanum lycopersicum L.) are nutrient-rich fruits with a high glutamic acid content, making them a suitable source for producing γ-aminobutyric acid (GABA) through the action of lactic acid bacteria (LAB). LAB plays a pivotal role in lactic fermentation, enhancing the taste and nutritional value of food products, often contributing to their health benefits. This study isolated GABA-producing LAB from tomatoes and assessed their potential for producing GABA-enriched fermented tomato juice. The results indicated the isolation of 15 LAB strains from the samples of five different tomatoes. All of these strains demonstrated the capability to produce GABA, with levels ranging from 1.732 to 3.113 mg/mL, as determined using thin-layer chromatography (TLC). Through sequencing, the promising strain TO42, identified as Weissella cibaria, was selected for tomato juice fermentation. Furthermore, GABA-enriched tomato juice was successfully fermented at 15 °Brix for 72 h, resulting in the highest recorded GABA and lactic acid content of 9.185 ± 0.398 mg/mL and 13.05 ± 1.56 g/L, respectively.

O tomate (Solanum lycopersicum L.) é uma fruta rica em nutrientes e com alto teor de ácido glutâmico, tornando-o uma fonte adequada para a produção de ácido γ-aminobutírico (GABA) através da ação de bactérias ácido lácticas (BAL). As BAL desempenham um papel fundamental na fermentação láctica, melhorando o sabor e o valor nutricional dos produtos alimentares, contribuindo frequentemente para os seus benefícios para a saúde. Este estudo teve como objetivo isolar BAL produtoras de GABA de tomates e avaliar seu potencial para a produção de suco de tomate fermentado enriquecido com GABA. Os resultados indicam o isolamento de 15 cepas de BAL a partir de amostras de cinco tomates diferentes. Todas essas cepas demonstraram capacidade de produzir GABA, com níveis variando de 1,732 a 3,113 mg/mL, conforme determinado por cromatografia em camada delgada (CCD). Através de sequenciamento, a cepa promissora TO42, identificada como Weissella cibaria, foi selecionada para fermentação de suco de tomate. Além disso, o suco de tomate enriquecido com GABA foi fermentado com sucesso a 15 °Brix por 72 h, resultando no maior teor registrado de GABA e ácido láctico de 9.185 ± 0.398 mg/mL e 13.05 ± 1.56 g/L, respectivamente.

Resumo

Abstract Pseudomonas fluorescens is one of the main causes of septicemic diseases among freshwater fish, causing severe economic losses and decreasing farm efficiency. Thus, this research was aimed to investigate the occurrence of P. fluorescens in Nile Tilapia (O. niloticus) fish in Egypt, gene sequencing of 16SrDNA gene, and antimicrobial susceptibility. P. fluorescens strains were detected in 32% (128/400) of apparently healthy (9%; 36/400) and diseased (23%; 92/400) Nile tilapia fish. The highest prevalence was observed in gills of fish, 31.3% followed by intestine 26.9%, liver 24.2%, and kidneys 17.6%. The PCR results for the 16SrDNA gene of P. fluorescens showed 16SrDNA gene in 30% of examined isolates. Moreover, Homogeny and a strong relationship between strains of P. fluorescens was confirmed using 16SrDNA sequences. Beside the responsibility of 16SrDNA gene on the virulence of P. fluorescens. The results of antimicrobial susceptibility tests revealed that all strains were resistant to piperacillin (100%), followed by ceftazidime (29.7%), and cefepime (25.8%). The strains of P. fluorescence were highly sensitive to cefotaxime (74.2%), followed by ceftriaxone and levofloxacin (70.3% each). Interestingly, 29.7% of strains of P. fluorescens were multiple antimicrobial-resistant (MAR).

Resumo Pseudomonas fluorescens é uma das principais causas de doenças septicêmicas em peixes de água doce, causando graves perdas econômicas e diminuindo a eficiência da fazenda. Assim, esta pesquisa teve como objetivo investigar a ocorrência de P. fluorescens em peixes de tilápia-do-nilo (O. niloticus) no Egito, sequenciamento do gene 16S rDNA e suscetibilidade antimicrobiana. Cepas de P. fluorescens foram detectadas em 32% (128/400) de peixes tilápia-do-nilo aparentemente saudáveis (9%; 36/400) e doentes (23%; 92/400). A maior prevalência foi observada nas brânquias dos peixes, 31,3%, seguida pelo intestino 26,9%, fígado 24,2% e rins 17,6%. Os resultados da PCR para o gene 16SrDNA de P. fluorescens mostraram o gene 16SrDNA em 30% dos isolados examinados. Além disso, a homogeneidade e uma forte relação entre cepas de P. fluorescens foi confirmada usando sequências de 16SrDNA. Além da responsabilidade do gene 16SrDNA na virulência de P. fluorescens. Os resultados dos testes de suscetibilidade antimicrobiana revelaram que todas as cepas foram resistentes à piperacilina (100%), seguida pela ceftazidima (29,7%) e cefepima (25,8%). As cepas de P. fluorescens foram altamente sensíveis à cefotaxima (74,2%), seguida pela ceftriaxona e levofloxacina (70,3% cada). Curiosamente, 29,7% das cepas de P. fluorescens eram multirresistentes a antimicrobianos (MAR).

Resumo

The current study describes the presence of Bacillus cereus (B. cereus) in contaminated foods of animal source and ready for human consumption with highlighting on their virulence contributing factors by detection of its virulence genes in addition to identification of their sequencing. Three hundred sixty food samples categorized as (228) meat products and (132) milk products were examined for B. cereus isolation and all of these isolates were confirmed by biochemical tests. Eighteen strains obtained from different food samples were examined for the attendance of a number of virulence genes (nheA, cytK, entFM, bceT and hblC genes) using uniplex PCR method. Furthermore, the B. cereus strains were valued for the sequencing of described genes. Generally 24.44% (88/360) food samples classified as 11.11% (40/360) meat products and 13.33% (48/360) milk products carried B. cereus according to cultural and biochemical properties, with geometric mean (1.5×107±0.15 CFU/g or mL) . The highest counts (above 105 CFU/g or mL) were originated from milk products (with geometric mean 2.2×107±0.22 CFU/g or mL) more than meat products (with geometric mean 1×107±0.19 CFU/g or mL). The results revealed that all of our isolates had one or more virulence (enterotoxin) genes. In our research, the most predominant genes were nheA (100%), followed by cytK (61.11%), entFM (33.33%), bceT (11.11%) then hblC (5.56%). Molecular method detected that overall, 5 strains (27.78%) harbored only 1 gene (nheA), 7 strains (38.88%) harbored 2 genes which classified as 5 strains (27.78%) (nheA and cytK), 2 strains (11.11%) have (nheA and entFM). Moreover, 5 strains (27.78%) have 3 genes classified as 3 strains (16.67%) harbored (nheA, cytK and entFM), 1 strain (5.56%) had (nheA, cytK and hblC), and 1 strain (5.56%) had (nheA, cytK and bceT). Only 1 strain (5.56%) carried 4 tested virulence genes (nheA, cytK, entFM and bceT) genes. The most prevalent gene in meat and dairy foods was nheA (100%). The nucleotide sequences of (bceT, cytK, entFM, hblC and nheA genes) of B. cereus strains were deposited in GenBank under accession no. (MW911824, MW911825, MW911826, MW911827 and MW911828), respectively. Our study was established to indicate the presence of virulent B. cereus in meat and milk products ready for human consumption as a result of deficient hygienic actions. So, a plain for good hygienic measures should be modified to avoid causing serious health problems to human due to ingestion of such products.

O presente estudo descreve a presença de Bacillus cereus em alimentos contaminados de origem animal e prontos para consumo humano, com destaque para seus fatores de contribuição de virulência por meio da detecção de seus genes de virulência, além da identificação de seu sequenciamento. Trezentas e sessenta amostras de alimentos categorizados como produtos cárneos (228) e produtos lácteos (132) foram examinadas para isolamento de B. cereus, e todos esses isolados foram confirmados por testes bioquímicos. Dezoito cepas obtidas de diferentes amostras de alimentos foram examinadas para a presença de uma série de genes de virulência (genes nheA, cytK, entFM, bceT e hblC) usando o método de PCR uniplex. Além disso, as cepas de B. cereus foram avaliadas para o sequenciamento dos genes descritos. De forma geral, 24,44% (88/360) das amostras de alimentos classificados como produtos cárneos (11,11%; 40/360) e produtos lácteos (13,33%; 48/360) transportavam B. cereus, de acordo com as propriedades culturais e bioquímicas, com média geométrica de 1,5 × 10 7 ± 0,15 CFU/g ou mL. Os resultados revelaram que todos os nossos isolados tinham um ou mais genes de virulência (enterotoxina). Em nossa pesquisa, os genes mais predominantes foram nheA (100%), seguidos de cytK (61,11%), entFM (33,33%), bceT (11,11%) e hblC (5,56%). O método molecular detectou que, no geral, 5 cepas (27,78%) apresentavam apenas 1 gene (nheA) e 7 cepas (38,88%) continham 2 genes que foram classificados como 5 cepas (27,78%) (nheA e cytK), 2 cepas (11,11%) possuíam (nheA e entFM). Além disso, 5 cepas (27,78%) continham 3 genes classificados como 3 cepas (16,67%) hospedados (nheA, cytK e entFM), 1 cepa (5,56%) tinha (nheA, cytK e hblC) e 1 cepa (5,56%) teve (nheA, cytK e bceT). Apenas 1 cepa (5,56%) carregava 4 genes de virulência testados (nheA, cytK, entFM e bceT). As sequências de nucleotídeos (genes bceT, cytK, entFM, hblC e nheA) de cepas de B. cereus foram depositadas no GenBank sob o número de acesso (MW911824, MW911825, MW911826, MW911827 e MW911828), respectivamente. Nosso estudo foi estabelecido para indicar a virulência de B. cereus em carnes e produtos lácteos prontos para consumo humano como resultado de ações higiênicas deficientes. Portanto, deve ser estabelecido um plano com boas medidas de higiene para evitar sérios problemas de saúde humana por causa da ingestão de tais produtos.

Resumo

Seventy-five pectinolytic strains collected from vegetables grown in the counties of Santarém, Belterra, and Mojuí dos Campos, located in the western region of the Pará State, Brazil, were studied according to their pathological and genetic variability. The strains were grouped in 5 clusters according to pathogenicity in potato, pepper, carrot, and onion, and 38 strains were selected for genetic analysis using rep-PCR. These strains were divided into 35 genetic groups according to rep-PCR at 70% similarity. These results indicated high pathological and genetic variability of the strains causing soft rot in vegetables in the western region of the Pará State, which will be used in etiological research and for the development and assessment of management techniques for the soft rot in vegetables in this region.

Setenta e cinco isolados de bactérias pectinolíticas coletados de hortaliças cultivadas nos municípios de Santarém, Belterra e Mojuí dos Campos, região oeste do Estado do Pará, Brasil, foram estudadas de acordo com sua variabilidade patológica e genética. Os isolados foram agrupados em cinco grupos de acordo com a patogenicidade em batata, pimenta, cenoura e cebola, e 38 isolados foram selecionados para análise genética por rep-PCR. Esses isolados foram divididos em 35 grupos genéticos de acordo com rep-PCR a 70% de similaridade. Esses resultados indicaram alta variabilidade patológica e genética dos isolados causadores da podridão mole em hortaliças da região oeste do Pará, os quais serão utilizados em pesquisas etiológicas e para o desenvolvimento e avaliação de técnicas de manejo da podridão mole em hortaliças nesta região.

Resumo

Oxidative stress is the result of an imbalance between the production of oxidant precursors and the capacity of antioxidant defense. Oxygen free radicals play an important role in causing diseases. In this study, the protective effect of ethanolic avocado on apoptosis caused by oxidative damage in the tissue of albino rats was investigated. 24 male albino rats of the Faculty of Veterinary Medicine in Mosul, Iraq, which were kept in standard conditions for at least 10 days before and through the experimental work, were examined. Four groups of rats include the control group (healthy group), the group of male rats with ethanolic avocado consumption; The third group of male rats that were treated with 0.5% of hydrogen peroxide H2O2; and the fourth group of male rats that were treated with both 0.5% H2O2 and avocado ethanolic extract (50 mg kg-1 BW) for four weeks. After fixing the tissues of the liver, kidney, lung, spleen and testis in 10% buffered formalin, they were stained with hematoxylin. TUNEL assay was performed using the TUNEL cell death assay kit to detect apoptotic cells. In this investigation, the histology results in four groups of rats showed that in the rats that were treated with avocado, there were minor tissue changes in their liver, kidney, and intestine, and the tissues of these organs were healthy. In TUNEL staining, it was also shown that there are no apoptotic cells in the liver, kidney and testis cells in avocado-treated rats. The results showed that ethanolic Avocado is useful against oxidative stress damage and it may be used to protect tissues against oxidative stress.(AU)

Resumo

Abstract The impact of antibiotics on growth, cocoon production was assessed in addition to isolation and characterization of bacteria associated with silkworm gut of infected larvae. Larval rearing was maintained at recommended conditions of temperature and humidity. Silkworm larvae showing abnormal symptoms were collected from the control group and dissected for gut collection. Bacteria were isolated from the gut content by spreading on agar plates and incubated at 37 °C for 48 hrs. Bacterial identification and phylogenetic analysis were carried out by 16S rRNA gene sequencing. The isolated bacteria were subjected to antimicrobial susceptibility test (disc diffusion methods) by using Penicillin (10 µg/mL), Tetracycline (30 µg/mL), Amoxicillin (25 µg/mL), Ampicillin (10 µg/mL), and Erythromycin (15 µg/mL). All isolated strains showed positive results for the catalase test. We isolated and identified bacterial strains (n = 06) from the gut of healthy and diseased silkworm larvae. Based on the 16S rRNA gene sequence, isolated bacteria showed close relation with Serratia, Bacillus, and Pseudomonas spp. Notably, 83.3% of strains were resistant to Penicillin, Tetracycline, Amoxicillin, Ampicillin, and Erythromycin but 16.6% showed antibiotic susceptibility to the above-mentioned commonly used antibiotics. Silkworm larvae fed on penicillin-treated leaves showed significant improvement in larval weight, larval length, and cocoon production. Significantly higher larval weight (6.88g), larval length (5.84cm), and cocoon weight (1.33g) were recorded for larvae fed on leaves treated with penicillin as compared to other antibiotics. Isolated bacterial strains showed close relation with Serratia spp., Bacillus spp. and Pseudomonas spp.

Resumo O impacto dos antibióticos no crescimento e na produção do casulo foi avaliado, além do isolamento e caracterização das bactérias associadas ao intestino de larvas infectadas do bicho-da-seda. A criação das larvas foi mantida nas condições recomendadas de temperatura e umidade. As larvas do bicho-da-seda com sintomas anormais foram coletadas do grupo controle e dissecadas para coleta do intestino. As bactérias foram isoladas do conteúdo intestinal por espalhamento em placas de ágar e incubadas a 37° C durante 48 horas. A identificação bacteriana e a análise filogenética foram realizadas pelo sequenciamento do gene 16S rRNA. As bactérias isoladas foram submetidas a teste de sensibilidade antimicrobiana (métodos de difusão em disco) com penicilina (10 µg / mL), tetraciclina (30 µg / mL), amoxicilina (25 µg / mL), ampicilina (10 µg / mL) e eritromicina (15 µg / mL). Todas as cepas isoladas apresentaram resultados positivos para o teste da catalase. Isolamos e identificamos cepas bacterianas (n = 06) do intestino de larvas de bicho-da-seda saudáveis e doentes. Com base na sequência do gene 16S rRNA, as bactérias isoladas mostraram estreita relação com Serratia, Bacillus e Pseudomonas spp. Notavelmente, 83,3% das cepas eram resistentes a penicilina, tetraciclina, amoxicilina, ampicilina e eritromicina, mas 16,6% mostraram suscetibilidade aos antibióticos comumente usados mencionados acima. As larvas do bicho-da-seda alimentadas com folhas tratadas com penicilina apresentaram melhora significativa no peso larval, comprimento larval e produção de casulo. Peso larval significativamente maior (6,88g), comprimento larval (5,84cm) e peso do casulo (1,33g) foram registrados para larvas alimentadas com folhas tratadas com penicilina, em comparação com outros antibióticos. Cepas bacterianas isoladas mostraram estreita relação com Serratia spp., Bacillus spp. e Pseudomonas spp.

Resumo

The research evaluated the isolation, identification, and prospection of species-specific beneficial bacteria in fish farming of ornamental fish Betta splendens. For this, the microbiological material was obtained from the intestinal tract of healthy specimens, with bacterial growth in selective culture medium Man Rogosa Sharped (MRS). Sixteen strains were isolated based on the response of in vitro tests of catalase, Gram, nilin blue, hemolytic activity and antibiogram to pathogens Aeromonas hydrophila, Pseudomonas aeroginosa, Enterococcus durans and Escherichia coli. Of the isolated strains C1BS and C5BS, they showed the best responses, which were later identified by the Maldi-TOF method as Lactobacillus plantarum and Enterococcus faecium. Due to the performance of lactic acid strains in in vitro tests and the bibliographic record of their performance as probiotics, the species have great potential for species-specific use in the ornamental production of Betta splendens.

A pesquisa avaliou o isolamento, a identificação e a prospecção de bactérias benéficas espécie-específico na piscicultura do peixe ornamental Betta splendens. Para isso, o material microbiológico foi obtido do trato intestinal de espécimes sadios, com crescimento bacteriano em meio de cultura seletivo Man Rogosa Sharped (MRS). Dezesseis cepas foram isoladas com base na resposta dos testes in vitro de catalase, Gram, azul de anilina, atividade hemolítica e antibiograma aos patógenos Aeromonas hydrophila, Pseudomonas aeroginosa, Enterococcus durans e Escherichia coli. Das cepas isoladas, C1BS e C5BS foram as que apresentaram melhores respostas, as quais posteriormente foram identificadas, pelo método Maldi-TOF, como Lactobacillus plantarum e Enterococcus faecium. Devido ao registro bibliográfico e ao desempenho das cepas ácido lácticas nos testes in vitro, essas apresentam um grande potencial para uso probiótico espécie-específico na produção ornamental de Betta splendens.

Resumo

Abstract In the current context of emerging drug-resistant fungal pathogens such as Candida albicans and Candida parapsilosis, discovery of new antifungal agents is an urgent matter. This research aimed to evaluate the antifungal potential of 2-chloro-N-phenylacetamide against fluconazole-resistant clinical strains of C. albicans and C. parapsilosis. The antifungal activity of 2-chloro-N-phenylacetamide was evaluated in vitro by the determination of the minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), inhibition of biofilm formation and its rupture, sorbitol and ergosterol assays, and association between this molecule and common antifungal drugs, amphotericin B and fluconazole. The test product inhibited all strains of C. albicans and C. parapsilosis, with a MIC ranging from 128 to 256 µg.mL-1, and a MFC of 512-1,024 µg.mL-1. It also inhibited up to 92% of biofilm formation and rupture of up to 87% of preformed biofilm. 2-chloro-N-phenylacetamide did not promote antifungal activity through binding to cellular membrane ergosterol nor it damages the fungal cell wall. Antagonism was observed when combining this substance with amphotericin B and fluconazole. The substance exhibited significant antifungal activity by inhibiting both planktonic cells and biofilm of fluconazole-resistant strains. Its combination with other antifungals should be avoided and its mechanism of action remains to be established.

Resumo No atual contexto de patógenos fúngicos resistentes emergentes tais como Candida albicans e Candida parapsilosis, a descoberta de novos agentes antifúngicos é uma questão urgente. Esta pesquisa teve como objetivo avaliar o potencial antifúngico da 2-cloro-N-fenilacetamida contra cepas clínicas de C. albicans e C. parapsilosis resistentes a fluconazol. A atividade antifúngica da substância foi avaliada in vitro através da determinação da concentração inibitória mínima (CIM), concentração fungicida mínima (CFM), ruptura e inibição da formação de biofilme, ensaios de sorbitol e ergosterol, e associação entre esta molécula e antifúngicos comuns, anfotericina B e fluconazol. O produto teste inibiu todas as cepas de C. albicans e C. parapsilosis, com uma CIM variando de 128 a 256 µg.mL-1, e uma CFM de 512-1,024 µg.mL-1. Também inibiu até 92% da formação de biofilme e causou a ruptura de até 87% de biofilme pré-formado. A 2-cloro-N-fenilacetamida não promoveu atividade antifúngica pela ligação ao ergosterol da membrana celular fúngica, tampouco danificou a parede celular. Antagonismo foi observado ao combinar esta substância com anfotericina B e fluconazol. A substância exibiu atividade antifúngica significativa ao inibir tanto as células planctônicas quanto o biofilme das cepas resistentes ao fluconazol. Sua combinação com outros antifúngicos deve ser evitada e seu mecanismo de ação deve ser estabelecido.

Resumo

Salmonella spp. is one of the major bacterial causes of foodborne gastroenteritis in humans. The aim of this study was to investigate antimicrobial susceptibility to cephalosporins and quinolones, and to identify the genetic mechanisms related to this resistance in strains of Salmonella spp. Seventy chicken carcass samples were collected from slaughterhouses in the state of Rio de Janeiro, Brazil. The phenotypic profile was detected by the disk-diffusion method and the search for genes encoding betalactamases, and resistance to quinolones was evaluated by PCR. The search for mutations in gyrA and parC was carried out by sequencing these genes. Eleven strains of Salmonella spp. of different serotypes were isolated. All the strains were resistant to at least one of the antimicrobials tested, and 63.64% (7/11) showed resistance to three or more antimicrobials. In the phenotypic test for ESBL production, 36.36% (4/11) of the strains were considered positive. PCR detected the resistance genes bla CMY-2, qnrB, bla CTX-M, and bla TEM. Among the isolates, 45.45% (5/11) simultaneously presented the bla CTX-M, bla TEM, qnrB genes and a mutation (Thr-57→Ser) in parC. Point mutations in the parC gene were detected in all the analyzed samples. Genes such as bla SHV, qnrA, qnrC, qnrD, qnrS, aac(6')-Ib, qepA, and oqxAB were not detected. The study identified Salmonella spp. resistant to cephalosporins and quinolones, with resistance genes and mutations in parC, highlighting concerns about the adoption of biosecurity measures, responsible use of antimicrobials, and surveillance of resistant strains in the poultry chain.(AU)

Resumo

Pseudomonas fluorescens is one of the main causes of septicemic diseases among freshwater fish, causing severe economic losses and decreasing farm efficiency. Thus, this research was aimed to investigate the occurrence of P. fluorescens in Nile Tilapia (O. niloticus) fish in Egypt, gene sequencing of 16SrDNA gene, and antimicrobial susceptibility. P. fluorescens strains were detected in 32% (128/400) of apparently healthy (9%; 36/400) and diseased (23%; 92/400) Nile tilapia fish. The highest prevalence was observed in gills of fish, 31.3% followed by intestine 26.9%, liver 24.2%, and kidneys 17.6%. The PCR results for the 16SrDNA gene of P. fluorescens showed 16SrDNA gene in 30% of examined isolates. Moreover, Homogeny and a strong relationship between strains of P. fluorescens was confirmed using 16SrDNA sequences. Beside the responsibility of 16SrDNA gene on the virulence of P. fluorescens. The results of antimicrobial susceptibility tests revealed that all strains were resistant to piperacillin (100%), followed by ceftazidime (29.7%), and cefepime (25.8%). The strains of P. fluorescence were highly sensitive to cefotaxime (74.2%), followed by ceftriaxone and levofloxacin (70.3% each). Interestingly, 29.7% of strains of P. fluorescens were multiple antimicrobial-resistant (MAR).

Pseudomonas fluorescens é uma das principais causas de doenças septicêmicas em peixes de água doce, causando graves perdas econômicas e diminuindo a eficiência da fazenda. Assim, esta pesquisa teve como objetivo investigar a ocorrência de P. fluorescens em peixes de tilápia-do-nilo (O. niloticus) no Egito, sequenciamento do gene 16S rDNA e suscetibilidade antimicrobiana. Cepas de P. fluorescens foram detectadas em 32% (128/400) de peixes tilápia-do-nilo aparentemente saudáveis ​​(9%; 36/400) e doentes (23%; 92/400). A maior prevalência foi observada nas brânquias dos peixes, 31,3%, seguida pelo intestino 26,9%, fígado 24,2% e rins 17,6%. Os resultados da PCR para o gene 16SrDNA de P. fluorescens mostraram o gene 16SrDNA em 30% dos isolados examinados. Além disso, a homogeneidade e uma forte relação entre cepas de P. fluorescens foi confirmada usando sequências de 16SrDNA. Além da responsabilidade do gene 16SrDNA na virulência de P. fluorescens. Os resultados dos testes de suscetibilidade antimicrobiana revelaram que todas as cepas foram resistentes à piperacilina (100%), seguida pela ceftazidima (29,7%) e cefepima (25,8%). As cepas de P. fluorescens foram altamente sensíveis à cefotaxima (74,2%), seguida pela ceftriaxona e levofloxacina (70,3% cada). Curiosamente, 29,7% das cepas de P. fluorescens eram multirresistentes a antimicrobianos (MAR).

Resumo

This study assessed the effect of strain and enriched cage stocking density on performance, egg size distribution, egg quality, and welfare status in laying hens. Lohmann brown (LB) and Lohmann LSL Classic (LW) strains at 20 weeks of age were allotted to different cage stocking densities, with 1016, 762, or 610 cm2 of cage floor area per hen. Live body weight at the age at sexual maturity and at 52 weeks of age, feather condition, and some egg quality parameters differed between hen strains and among stocking densities (p<0.01; p<0.05). However, age at sexual maturity, livability, egg production, heterophil-to-lymphocyte ratio, and duration of tonic immobility were similar between hen strains and among stocking densities (p>0.05). Furthermore, egg size distribution was similar between hen strains (p>0.05), but dissimilar among stocking densities (p<0.01). Rectal and comb temperatures differed between hen strains and among stocking densities, respectively (p<0.01; p<0.05). The age of hens influenced egg quality variables (except for egg weight and shape index), feather condition, and body region temperatures (p<0.01). These results suggest that an enriched cage floor area of up to 610 cm2 per hen does not compromise production performance and welfare status, except for the body weight of laying hens. Additionally, the two strains might be at similar levels regarding overall performance and welfare status, excluding body weight and feather condition.(AU)

Resumo

Feathers make up 7% of the total weight of adult chickens and keratin protein makes up 85% of the feathers. Today, the keratinase enzymes of some Bacillus strains are used to degrade and process raw keratin waste for animal and poultry feed. According to various studies, the probiotic properties of some spore-shaped Bacillus have also been proven. The study aimed to isolation of the keratinolytic Bacillus bacteria that they have probiotic properties for using in the livestock and poultry feed industry. We were able to isolate 8 strains of Bacillus licheniformis with kreatin degrading properties from the soil of Baharan chicken slaughterhouse (Qom city, Iran) applying heat shock, alcohol- and keratin-rich culture medium, and after microscopic and biochemical analysis, 16S rDNA gene was isolated. The measurement results of keratinase activity showed that the three strains of Bacillus licheniformis pvkr6, pvkr 15, and pvkr41 had the highest activity with 124.08, 101.1, and 100.18 U/ml. The results of probiotic properties evaluation also revealed that among all the isolates, only Bacillus licheniformis pvkr15 and Bacillus licheniformis PTCC 1595 (positive control) were γ-hemolytic strains. The percentage of surface hydrophobicity of the strains was obtained from 3.27 to 30.57. It was also shown that, on average, all the strains had acceptable susceptibility to the tested antibiotics except penicillin G. Bacillus licheniformis pvkr15 with highest keratinase activity (101.1U/ml) was considered an optional probiotics due to its abilities such as (biofilm formation, being safe cause of γ-hemolytic activity, high susceptibility to antibiotics such as streptomycin, gentamicin, cefixime, amoxicillin, tetracycline, vancomycin, erythromycin and having a moderate hydrophilic (hydrophobicity: 19.09%), high survivability in pH 2, 2.5 and 3, strong resistance to bile salts and moderate antagonistic activity against pathogenic bacterium like Proteus mirabilis and the ability to grow under anaerobic conditions). By using this strain, after hydrolysis of keratin protein in the feather structure, to replace part of the protein of livestock and poultry feed, not only is no need to separate bacteria from the feed, but also the strain play role of an useful and effective additive in animal growth.

As penas representam 7% do peso total das galinhas adultas e a proteína de queratina compõe 85% das penas. Hoje, as enzimas queratinase de algumas cepas de Bacillus são usadas para degradar e processar resíduos de queratina brutos para alimentação de animais e aves. De acordo com vários estudos, as propriedades probióticas de alguns Bacillus em forma de esporos também foram comprovadas. O estudo teve como objetivo o isolamento das bactérias queratinolíticas Bacillus que possuem propriedades probióticas para uso na indústria de ração animal e avícola. Conseguimos isolar 8 cepas de Bacillus licheniformis com propriedades degradantes de creatina do solo do abatedouro de frangos de Baharan (cidade de Qom, Irã) aplicando choque térmico, meio de cultura rico em álcool e queratina e, após análise microscópica e bioquímica, o gene 16S rDNA foi isolado. Os resultados da medição da atividade da queratinase mostraram que as três cepas de Bacillus licheniformis pvkr6, pvkr15 e pvkr41 tiveram a maior atividade com 124,08, 101,1 e 100,18 U/ml. Os resultados da avaliação das propriedades probióticas também revelaram que dentre todos os isolados apenas Bacillus licheniformis pvkr15 e Bacillus licheniformis PTCC 1595 (controle positivo) eram cepas γ-hemolíticas. A porcentagem de hidrofobicidade superficial das cepas foi obtida de 3,27 a 30,57. Também foi demonstrado que, em média, todas as cepas apresentaram suscetibilidade aceitável aos antibióticos testados, exceto penicilina G. Bacillus licheniformis pvkr15 com maior atividade de queratinase (101,1U/ml) foi considerado um probiótico opcional devido às suas habilidades como formação de biofilme, sendo causa segura de atividade γ-hemolítica, alta suscetibilidade a antibióticos como estreptomicina, gentamicina, cefixima, amoxicilina, tetraciclina, vancomicina, eritromicina e ter uma hidrofílica moderada (hidrofobicidade: 19,09%), alta capacidade de sobrevivência em pH 2, 2,5 e 3, forte resistência aos sais biliares e atividade antagonista moderada contra bactérias patogênicas como Proteus mirabilis e a capacidade de crescer em condições anaeróbicas. Ao utilizar esta cepa, após a hidrólise da proteína queratina na estrutura da pena, para substituir parte da proteína da ração de gado e aves, não só não há necessidade de separar as bactérias da ração, mas também a cepa desempenha um papel útil e eficaz aditivo no crescimento animal.

Resumo

This work reviews the effect of environmental enrichments (perches, platforms, stocking density, outdoor access, bale, and dust bathing substrates) on the performance of fast and slow-growing commercial broiler strains. The performance of both slow and fast-growing commercial broiler strains under conventional production systems are generally poor, especially regarding the welfare status. One of the strategies to improve the performance of commercial broiler strains is by adding enrichment objects to production systems. The addition of enrichments to production systems should improve animal welfare, have no negative effect on production performance, and be both economically practicable and feasible to employ. Perches and platforms are the most common enrichments used to increase the activity of broiler chickens to improve leg conditions. The use of perches and platforms could lead to the reduction in the incidence of footpad dermatitis, hockburns and breast blisters, with subsequent effects on meat quality. Moreover, the provision of outdoor access could improve the biology responses of broiler chickens to various environmental stimuli, with a profound effect on performance and meat quality traits. Furthermore, another enrichment strategies that could increase the exploratory behavior and the general welfare of broiler chickens is the use of dustbathing and bale subtrates. Moreover, adjusting the stocking density provides broiler chickens with the necessary space for movement, reduces crowding, trampling and the associated agonistic behavior. However, the effect of some of these enrichments (perches, platform, bale) objects may vary depending on height, age, sex, and strain of the chickens.(AU)

Resumo

Abstract Due to extensive application of antibiotics as growth promoters in animal feed, antimicrobial resistance has been increased. To overcome this challenge, rumen microbiologists search for new probiotics to improve the rate of livestock production. The present study was aimed to isolate and evaluate breed-specific lactic acid bacteria (LAB) as potential animal probiotics. The current study was conducted during 10 months from July 2020 to April 2021, in which a total of n=12 strains were isolated from different samples including milk, rumen, and feces of Nilli Ravi Buffaloes. These isolates were evaluated for their antimicrobial potential against common animal pathogens (Bacillus spp., E. coli, Staphylococcus aureus, Salmonella spp., Listeria spp.). All the isolates were identified using 16S rRNA gene sequencing and the phylogenetic analyses inferred that these strains showed close relations to the species of various genera; Enterococcus lactis, Pediococcus pentosaceus, Bacillus subtilis Weissella cibaria, Weissella soli, Bacillus tequilensis, Weissella bombi, Bacillus licheniformis, Lactococcus lactis, Bacillus megaterium, Lactobacillus ruminis, and Lactococcus lactis. NMCC-Ru2 has exhibited the enormous potential of antimicrobial activity, 28 mm, for Salmonella typhimurium;23 mm for Listeria monocytogenes 21 mm for E.coil. Highest resistance was seen in NMCC-Ru2 agasint test antbiotic, like 25.5 mm for Tetracycline. Overall results revesl that the probiotic profile of isolates was achieved using standard criteria, particularly with animal probiotic properties

Resumo Devido à extensa aplicação de antibióticos como promotores de crescimento na alimentação animal, a resistência aos antimicrobianos aumentou. Para superar esse desafio, os microbiologistas do rúmen buscam novos probióticos para melhorar a produtividade do gado. O presente estudo teve como objetivo isolar e avaliar bactérias lácticas específicas de raças (BAL) como potenciais probióticos animais. 12 cepas foram isoladas de diferentes amostras, incluindo leite, rúmen e fezes de búfalos Nilli Ravi. Esses isolados foram avaliados quanto ao seu potencial antimicrobiano contra patógenos animais comuns (Bacillus spp., E. coli, Staphylococcus aureus, Salmonella spp., Listeria spp.). Todos os isolados foram identificados por meio do sequenciamento do gene 16S rRNA e as análises filogenéticas inferiram que essas cepas apresentaram estreita relação com as espécies de vários gêneros; Enterococcus lactis, Pediococcus pentosaceus, Bacillus subtilis, Weissella cibaria, Weissella soli, Bacillus tequilensis, Weissella bombi, Bacillus licheniformis, Lactococcus lactis, Bacillus megaterium, Lactobacillus ruminis e Lactococcus lactis. O perfil probiótico dos isolados foi obtido usando critérios padrão, particularmente com propriedades probióticas animais.

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Antibiotic resistance in bacteria is the primary challenge for health worldwide. The widespread of food poisoning due to use of stored food items increasing day by day. The present study was designed to research the protective effect of Eucalyptus camaldulensis methanolic extract again selected strains of bacteria (Escherichia coli, Bacillus cereus, Bacillus subtilis and Staphylococcus aureus). The treatment was used for several concentrations and then characterized on the basis of temperature and pH variations. Results were observed by MIC (minimum inhibitory concentration) against these strains. It was noted that maximum inhibition of E. camaldulensis against E.coli (5.6mm) was observed at 250 mg/mL while it was 3.6mm against B. cereus at 200 mg/mL, Staph. aureus showed maximum (3.1mm) zone at 250 mg/mL. at variable temperature of E. camaldulensis extract, it was observed that MIC of B. cereus was 6.4mm at 80 °C. For other strains the results revealed that the maximum zone against B. subtilis was 5.7mm at 121 °C and for Staph. aureus it was 6.6mm at 80 °C. By observing the results by changing pH it was observed that MIC produced against B. cereus was 6.2mm at 7pH, against B. subtilis zone of inhibition was 7.2mm at 5pH, for E.coli it was 5.4mm at 3pH. Means of all the variable results of different (concentration, pH and temperature) were compared by using one way ANOVA. The current study suggested that the methanolic extract of E. camaldulensis was found effective in control of antibiotic resistant strains and this study strengthens the fact of using herbal solutions for control of antibiotic resistant bacterial infections.

A resistência bacteriana a antibióticos é o principal desafio para a saúde em todo o mundo. A disseminação da intoxicação alimentar devido ao uso de alimentos armazenados aumenta a cada dia. O presente estudo foi elaborado para pesquisar o efeito protetor do extrato metanólico de Eucalyptus camaldulensis em cepas selecionadas de bactérias (Escherichia coli, Bacillus cereus, Bacillus subtilis e Staphylococcus aureus). O tratamento foi aplicado em várias concentrações e então caracterizado com base nas variações de temperatura e pH. Os resultados foram observados por MIC (concentração inibitória mínima) contra essas cepas. Foi notado que a inibição máxima de E. camaldulensis contra E. coli (5,6 mm) ocorreu na concentração de 250 mg/mL, enquanto foi de 3,6 mm contra B. cereus a 200 mg/mL, e Staph. aureus apresentou zona máxima (3,1 mm) a 250 mg/mL. Em temperatura variável do extrato de E. camaldulensis, observou-se que a MIC de B. cereus foi de 6,4 mm a 80 °C. Para outras cepas, os resultados revelaram que a zona máxima contra B. subtilis foi de 5,7 mm a 121 °C, e para Staph. aureus foi de 6,6 mm a 80 °C. Observando os resultados pela alteração do pH, observou-se que a MIC produzida contra B. cereus foi de 6,2 mm a 7 pH, contra B. subtilis a zona de inibição foi de 7,2 mm a 5 pH, para E. coli foi de 5,4 mm a 3 pH. As médias de todos os resultados variáveis ​​ (concentração, pH e temperatura) foram comparadas usando ANOVA unidirecional. O presente estudo sugeriu que o extrato metanólico de E. camaldulensis foi considerado eficaz no controle de cepas resistentes a antibióticos, e este estudo reforça o uso de soluções fitoterápicas para o controle de infecções bacterianas resistentes a antibióticos.

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The current research was designed to reach extracellular protease production potential in different strains of Sordaria fimicola which were previously obtained from Dr. Lamb (Imperial College, London) from North Facing Slope and South Facing Slope of Evolution Canyon. After initial and secondary screening, two hyper-producers strains S2 and N6 were selected for submerged fermentation and cultural conditions including temperature, pH, incubation period, inoculum size, substrate concentration, and different carbon and nitrogen sources were optimized for enzyme production. S2 strain showed maximum protease production of 3.291 U/mL after 14 days of incubation at 30 °C with 7 pH, 1% substrate concentration and 1 mL inoculum, While N6 strain showed maximum protease production of 1.929 U/mL under fermentation optimized conditions. Another aim of the present research was to underpin the biodiversity of genetics and post-translational modifications (PTMs) of protease DPAP (peptidyl-aminopeptidase) in Sordaria fimicola. Five polymorphic sites were observed in amino acid sequence of S. fimicola strains with reference to Neurospora crassa. PTMs prediction from bioinformatics tools predicted 38 phosphorylation sites on serine residues for protease peptidyl-aminopeptidase in S1 strain of S. fimicola while 45 phosphorylation sites on serine in N7 strain and 47 serine phosphorylation modifications were predicted in N. crassa. Current research gave an insight that change in genetic makeup effected PTMs which ultimately affected the production of protease enzyme in different strains of same organism (S. fimicola). The production and molecular data of the research revealed that environmental stress has strong effects on the specific genes through mutations which may cause genetic diversity. S. fimicola is non- pathogenic fungus and has a short life cycle. This fungus can be chosen to produce protease enzyme on a commercial scale.

A pesquisa atual foi projetada para alcançar o potencial de produção de protease extracelular em diferentes cepas de Sordaria fimicola que foram previamente obtidas do Dr. Lamb (Imperial College, Londres) de North Facing Slope e South Facing Slope de Evolution Canyon. Após a triagem inicial e secundária, duas cepas hiperprodutoras S2 e N6 foram selecionadas para fermentação submersa e condições culturais, incluindo temperatura, pH, período de incubação, tamanho do inóculo, concentração de substrato, e diferentes fontes de carbono e nitrogênio foram otimizadas para produção de enzima. A cepa S2 apresentou produção máxima de protease de 3,291 U/mL após 14 dias de incubação a 30 °C com pH 7, concentração de substrato de 1% e inóculo de 1 mL, enquanto a cepa N6 apresentou produção máxima de protease de 1,929 U/mL em condições otimizadas de fermentação. Outro objetivo da presente pesquisa foi sustentar a biodiversidade da genética e modificações pós-tradicionais (PTMs) da protease DPAP (peptidil-aminopeptidase) em Sordaria fimicola. Cinco sítios polimórficos foram observados na sequência de aminoácidos de cepas de S. fimicola com referência a Neurospora crassa. A previsão de PTMs a partir de ferramentas de bioinformática previu 38 locais de fosforilação em resíduos de serina para protease peptidil-aminopeptidase na cepa S1 de S. fimicola, enquanto 45 locais de fosforilação em serina na cepa N7 e 47 modificações de fosforilação de serina foram previstas em N. crassa. A pesquisa atual deu uma ideia de que a mudança na composição genética afetou os PTMs que, em última análise, afetaram a produção da enzima protease em diferentes cepas do mesmo organismo (S. fimicola). A produção e os dados moleculares da pesquisa revelaram que o estresse ambiental tem fortes efeitos sobre genes específicos por meio de mutações que podem causar diversidade genética. S. fimicola é um fungo não patogênico e tem um ciclo de vida curto. Esse fungo pode ser escolhido para produzir enzima protease em escala comercial.

Resumo

Over the past few decades, there has been increasing interest in using probiotics as an alternative to antibiotics. Researchers have conducted studies to investigate bacterial strains for their probiotic potential. Like other animals, fish also have several bacterial strains in their gut that possess probiotic properties, although this is limited to Bacillus species. Therefore, this study aimed to isolate and characterize probiotic Bacillus species from the gut of Masheer fish (Tor Puititora). Four pure bacterial isolates were selected as potential probiotic strains based on selection criteria, including survival rate in acid and bile salt. The isolates exhibited significant antimicrobial activity against pathogenic bacteria, including Escherichia coli (ATCC8739), Pseudomonas aeruginosa (ATCC9027), and Staphylococcus aureus (ATCC6538). MF2 and MF3 demonstrated clear zones, including antimicrobial activity against all three indicator pathogens. MF1 and MF4 exhibited antimicrobial activity against E. coli (ATCC8739) and P. aeruginosa (ATCC9027). Furthermore, the 16S rRNA gene sequences of all isolates exhibited a close association with Bacillus tequilensis (KCTC13622), with nucleotide similarity of 98.63%, 98.25%, 98.80%, and 98.35%, respectively. Our results demonstrate that these bacterial isolates show promise as an alternative to antibiotics in the fisheries food system. In this study, all isolates identified in the fish gut were subspecies of B. tequilensis.

Nas últimas décadas, tem havido um interesse crescente no uso de probióticos como uma alternativa aos antibióticos. Pesquisadores conduziram estudos para investigar cepas bacterianas quanto ao seu potencial probiótico. Como outros animais, os peixes também têm várias cepas bacterianas em seus intestinos que possuem propriedades probióticas, embora isso seja limitado a espécies de Bacillus. Portanto, o objetivo deste estudo foi isolar e caracterizar espécies probióticas de Bacillus do intestino de peixe Masheer (Tor Puititora). Quatro isolados bacterianos puros foram selecionados como potenciais cepas probióticas com base em critérios de seleção, incluindo taxa de sobrevivência em ácido e sal biliar. Os isolados exibiram atividade antimicrobiana significativa contra bactérias patogênicas, incluindo Escherichia coli (ATCC8739), Pseudomonas aeruginosa (ATCC9027) e Staphylococcus aureus (ATCC6538). MF2 e MF3 demonstraram zonas claras, incluindo atividade antimicrobiana contra todos os três patógenos indicadores. MF1 e MF4 exibiram atividade antimicrobiana contra E. coli (ATCC8739) e P. aeruginosa (ATCC9027). Além disso, as sequências do gene 16S rRNA de todos os isolados exibiram uma associação próxima com Bacillus tequilensis (KCTC13622), com similaridade de nucleotídeos de 98,63%, 98,25%, 98,80% e 98,35%, respectivamente. Nossos resultados demonstram que esses isolados bacterianos mostram-se promissores como uma alternativa aos antibióticos no sistema alimentar da pesca. Neste estudo, todos os isolados identificados no intestino dos peixes eram subespécies de B. tequilensis.

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Poultry and poultry products are considered the predominant sources of Salmonella enterica contamination and are important reservoirs of bacteria with antimicrobial resistance. The objective of this study was to identify Salmonella with multidrug resistance (MDR) phenotype, with the ability to form biofilms and elucidate the presence of genes that encode antimicrobial resistance in isolates from the broiler production chain in the state of Maranhão, Brazil. A total of 121 strains of S. enterica of different serovars were evaluated for antimicrobial susceptibility, and of these, 26 strains were used to detect the ability to form biofilms and identify resistance genes using PCR. Antimicrobial resistance was observed in 95 (78.5%) Salmonella isolates, and 57 (47.1%) showed MDR phenotype. The isolates showed greater resistance to the sulfonamide principles (58.7%), trimethoprim (48.8%), tetracycline (45.4%), nalidixic acid (44.6%), amoxicillin and ampicillin (26.4%), and cefazolin (22.3%). Salmonella Schwarzengrund (n=21/61.7%), Albany (n=15/62.5%), and Enteritidis (n=4/44.5%) showed the highest indices of MDR phenotype. The ability to form biofilms at 37°C was found in 13 of the 26 strains evaluated, which were considered poor producers. The resistance genes blaCTX-M, blaCTX-M2, blaSHV, sul1, sul2, tetA, tetB, tetC, tetE, dfrA12, and dfrA1 were observed in the serovars Schwarzengrund, Albany, Enteritidis, Heidelberg, and Typhimurium. The results showed a high occurrence of S. enterica, with multiple resistance to conventional antimicrobials and the ability to form biofilms in the poultry production chain.

As aves e os produtos de origem aviária são fontes de contaminação predominantes de Salmonella enterica e importantes reservatórios de bactérias com resistência antimicrobiana. Objetivou-se identificar Salmonella com fenótipos de multirresistência a drogas (MDR), com a capacidade de formação de biofilmes e a presença de genes que codificam resistência antimicrobiana em isolados da cadeia de frangos de corte, do estado Maranhão, Brasil. Avaliaram-se 121 cepas de S. enterica de sorovares diferentes quanto ao teste de suscetibilidade aos antimicrobianos e destas, 26 cepas para detecção da capacidade de formar biofilmes e genes de resistência pela técnica de PCR. Foram encontradas resistência antimicrobiana em 95 (78,5%) dos isolados de Salmonella e 57 (47,1%) apresentaram fenótipos MDR. Os isolados apresentaram maior resistência aos princípios sulfonamida (58,7%), trimetoprim (48,8%), tetraciclina (45,4%), ácido nalidíxico (44,6%), amoxicilina e ampicilina (26,4%) e cefazolin (22,3%). Os sorovares Salmonella Schwarzengrund (n=21/61.7%), Albany (n=15/62.5%) e Enteritidis (n=4/44.5%) apresentaram os maiores índices de fenótipos MDR. A capacidade de formar biofilmes foi encontrada em 13 cepas avaliadas, consideradas fracamente produtoras. Nos sorovares S. Schwarzengrund, Albany, Enteritidis, Heidelberg e Typhimurium foram detectados os genes de resistência blaCTX-M, blaCTX-M2, blaSHV, sul1, sul2, tetA, tetB, tetC, tetE, dfrA12 e dfrA1. Os resultados evidenciaram a elevada ocorrência de fenótipos de S. enterica com resistência múltipla a antimicrobianos convencionais, com capacidade de formar biofilmes, na cadeia produtiva de aves destinadas ao consumo humano.

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Proteobacteria comprising species of Pseudomonas syringae group cause diseases of many plants around the world. The phytopathogen has a complex taxonomic structure, which is constantly being revised due to the emergence of new molecular and biochemical diagnostic methods. Here for the first time, we describe the genetic and phenotypic diversity of 57 strains of Pseudomonas syringae isolated from affected soybeans, cereals, sunflowers, and other plants in the Russian Federation from 1950 to 2019. Genetic diversity was assessed by Multi Locus Sequence Analysis (MLSA) using fragments of the genes of glyceraldehyde-3-phosphate dehydrogenase (gapdh), the DNA-directed RNA polymerase subunit D (rpoD), gyrase (topoisomerase) B subunit (gyrB), and citrate synthase I (gltA). The synthesis of syringomycin and coronatine by bacteria was assessed by the reaction of susceptible yeast culture, seedlings of barley, tomato, and sunflower, and by presence of toxin genes confirmed by PCR test. The pathogenicity of the strains was confirmed on seedlings of dicotyledonous and monocotyledonous plants of peas, soybean, sunflowers, barley and wheat, as the most affected crops. The sensitivity of bacteria to 10 antibiotics of the main mechanisms of activity and two bactericidal commercial products was tested by standard disc method. The obtained results showed a high genetic homogeneity of the Russian population of P. syringae, which infects various agricultural crops, and an increase in the proportion of antibiotic-resistant strains over the years.

Proteobactérias compreendendo espécies do grupo Pseudomonas syringae causam doenças de muitas plantas ao redor do mundo. O fitopatógeno possui uma estrutura taxonômica complexa, que está em constante revisão devido ao surgimento de novos métodos de diagnóstico molecular e bioquímico. Aqui, pela primeira vez, descrevemos a diversidade genética e fenotípica de 57 cepas de Pseudomonas syringae isoladas de soja, cereais, girassol e outras plantas afetadas na Federação Russa de 1950 a 2019. A diversidade genética foi avaliada por análise de sequência multilocus (MLSA) usando fragmentos dos genes da gliceraldeído-3-fosfato desidrogenase (gapdh), a subunidade D da RNA polimerase dirigida por DNA (rpoD), a subunidade B da girase (topoisomerase) (gyrB) e a citrato sintase I (gltA). A síntese de siringomicina e coronatina por bactérias foi avaliada pela reação de cultura de leveduras suscetíveis, plântulas de cevada, tomate e girassol, e pela presença de genes de toxina confirmados pelo teste de PCR. A patogenicidade das cepas foi confirmada em mudas de plantas dicotiledôneas e monocotiledôneas de ervilha, soja, girassol, cevada e trigo, como as culturas mais afetadas. A sensibilidade das bactérias a 10 antibióticos dos principais mecanismos de atividade e dois produtos bactericidas comerciais foi testada pelo método de disco padrão. Os resultados obtidos mostraram uma alta homogeneidade genética da população russa de P. syringae, que infecta várias culturas agrícolas, e um aumento na proporção de cepas resistentes a antibióticos ao longo dos anos.

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The current study characterizes the genetic distribution of virulence and antimicrobial resistance of Streptococcus lutetiensis and Streptococcus equinus isolated from cows with clinical mastitis using whole genome sequencing (WGS). Although they are not the protagonist species within the genus Streptococcus, recent studies have isolated these species associated with bovine mastitis. In addition, these species are reported and isolated from humans and other animals. A total of four strains of S. lutetiensis and one of S. equinus were isolated from five cows with identified cases of clinical mastitis at a dairy farm near Ithaca, New York. Nineteen genes associated with antimicrobial resistance and 20 genes associated with virulence were identified in the analyzed strains. All strains presented genes associated with resistance: alr, ddl, gdpD, kasA, murA, lsa(E), msr(D), mef(A), gidB, and LiaF. Resistance genes associated with several different classes of antibiotics have also been reported. Sixteen virulence-associated genes were identified in all strains. Based on our findings, we conclude that the studied species have the potential to cause mastitis in cattle, and further studies are important to elucidate their role.

O presente estudo caracteriza a distribuição genética de virulência e resistência antimicrobiana de Streptococcus lutetiensis e Streptococcus equinus isolados de vacas com mastite clínica usando sequenciamento completo do genoma. Apesar de não serem as espécies protagonistas dentro do gênero Streptococcus, estudos recentes têm isolado essas espécies associadas à mastite bovina. Além disso, essas espécies são relatadas e isoladas de humanos e outros animais. Um total de quatro cepas de S. lutetiensis e uma de S. equinus foram isoladas de cinco vacas com casos identificados de mastite clínica em uma fazenda leiteira perto de Ithaca, Nova York. Dezenove genes associados à resistência antimicrobiana e 20 genes associados à virulência foram identificados nas cepas analisadas. Todas as linhagens apresentaram genes associados à resistência: alr, ddl, gdpD, kasA, murA, lsa(E), msr(D), mef(A), gidB e LiaF. Genes de resistência associados a várias classes diferentes de antibióticos também foram relatados. Dezesseis genes associados à virulência foram identificados em todas as cepas. Com base em nossos achados, concluímos que as espécies estudadas têm potencial para causar mastite em bovinos e mais estudos são importantes para elucidar seu papel.