Distinct carbapenems susceptibility profiles in isogenic isolates of Klebsiella pneumoniae presenting Ompk36 disruption and expression of down-regulated blaKPC-2 / Perfis distintos de suscetibilidade a carbapenêmicos em isolados isogênicos de Klebsiella pneumoniae apresentando interrupção de Ompk36 e expressão de blaKPC-2 negativamente regulada

The isolation of multidrug-resistant Klebsiella pneumoniae in hospitals is a major public health threat, increasing patient hospitalization costs, morbidity and mortality. Therefore, this work investigated the resistance mechanisms that produced different carbapenems susceptibility profiles in two isogenic strains of K. pneumoniae isolated from the same patient in a public hospital in Recife, Pernambuco. The genes that encode the main porins in K. pneumoniae, ompK35 and ompK36, and several beta-lactamase genes were analyzed. The expression of these genes was evaluated by quantitative real time PCR (polymerase chain reaction) with reverse transcriptase (RT-qPCR). SDS-PAGE (sodium dodecyl sulphate­polyacrylamide gel electrophoresis) was performed to analyze the outer membrane proteins. The analysis of the ompK36 genetic environment disclosed an IS903 insertion sequence disrupting this gene in the ertapenem resistant isolate (KPN133). The blaKPC-2 gene showed down-regulated expression in both isolates. Our findings show that changes in porins, especially OmpK36, are more determinant to carbapenems susceptibility profile of bacterial isolates than variations in blaKPC gene expression.

O isolamento de Klebsiella pneumoniae multirresistente em hospitais é uma grande ameaça à saúde pública, aumentando os custos de internação, morbidade e mortalidade dos pacientes. Portanto, este trabalho investigou os mecanismos de resistência que produziram diferentes perfis de suscetibilidade aos carbapenêmicos em duas cepas isogênicas de K. pneumoniae isoladas do mesmo paciente em um hospital público em Recife, Pernambuco. Foram analisados ​​os genes que codificam as principais porinas em K. pneumoniae, ompK35 e ompK36, e diversos genes de beta-lactamases. A expressão desses genes foi avaliada por PCR (reação em cadeia da polimerase quantitativa em tempo real) com transcriptase reversa (RT-qPCR). SDS-PAGE (dodecil sulfato de sódio-poliacrilamida gel eletroforese) foi realizada para analisar as proteínas da membrana externa. A análise do ambiente genético ompK36 revelou uma sequência de inserção IS903 interrompendo este gene no isolado resistente ao ertapenem (KPN133). O gene blaKPC-2 apresentou expressão negativamente regulada em ambos os isolados. Nossos achados mostram que alterações nas porinas, especialmente OmpK36, são mais determinantes no perfil de suscetibilidade aos carbapenêmicos de isolados bacterianos do que variações na expressão do gene blaKPC.

Chronic superficial infection in a dog caused by multidrug-resistant Pseudomonas aeruginosa

Background: Pseudomonas aeruginosa is a gram-negative aerobic bacterium and non-glucose fermenting, that usually causes opportunistic infections in animals, including humans. It is rarely involved in primary disease. The antibioticresistant bacterial strains are mainly developed due to the inappropriate use of antibiotics, however treating P. aeruginosa infections can be difficult owing to their natural resistance to antibiotics. Furthermorer resistant microorganisms such as P. aeruginosa grow by developing biofilms. Inaccurate diagnoses and absence of adequate microbiological tests can cause difficulties in resolving cases. This report describes a case of chronic superficial infection in a bitch caused by multidrugresistant Pseudomonas aeruginosa (MDR-PA). Case: A 6-year-old bitch Shih Tzu, initially presented with an exudative erythematous lesion in the snout region, which progressed to deep lesions, and spread to the back and limbs; furthermore, the animal always experienced a fever before new wounds emerged. Lesion samples, collected using a swab and processed at the Veterinary Microbiology Laboratory of the Federal University of Jatai (UFJ), revealed the presence of Pseudomonas aeruginosa. The isolate was multidrug-resistant and a carrier of TEM and ppyR genes. In the diffusion disk antibiogram, the isolate was found resistant to 14 different antibiotics belonging to 6 classes. Antimicrobial resistance was also tested using the minimum inhibitory concentration (MIC) test against imipenem, ceftazidime, ciprofloxacin, ticarcillin + clavulanic acid and aztreonam present in the MIC test strip. Treatment with amikacin and muporicin proved to be effective; however, owing to lesions extending to the face and palpebral involvement, the animal lost its eyeballs. Discussion: Pseudomonas aeruginosa is frequently associated with nosocomial infections mainly affecting immunosuppressed patients. Among the antibiotics tested, the group with the highest number of ineffective antibiotics was beta-lactams, where sensitivity was only observed for ticarcillin and ceftazidime. Recent studies have demonstrated that ceftazidime can reduce biofilm volume, inhibit motility, and repress the expression of genes associated with bacterial adhesion in P. aeruginosa. Therefore, the production of biofilm in P. aeruginosa is an important virulence factor as it facilitates a stable environment for the microorganism, which protects the bacteria from contact with antimicrobials. In addition, prolonged exposure to a wide variety of antimicrobials creates an environment of selective pressure between microorganisms, facilitating the emergence of multidrug-resistant strains. Furthermore, it is now well recognized that low doses of antibiotics, administered during continuous and fluctuating treatments, can stimulate biofilm establishment and are partly responsible for biofilm-specific antimicrobial tolerance. The resistance profile of P. aeruginosa isolated from dogs varies considerably, and the presence of isolates with a possible biofilm production capacity represents a challenge for the interpretation of the antimicrobial susceptibility profile. Culture and antibiogram is fundamentally important, both clinically and in environmental monitoring, in addition to the use of antibiogram data for decision making in clinical treatment.

Resistant enterococci isolated from raw sheep's milk and cheeses from South region of Brazil / Enterococos resistentes isolados de leite cru de ovelha e queijos da região Sul do Brasil

Enterococci have been used as sentinel organisms for monitoring antimicrobial resistance in food, humans, and other animals. In this sense, the present study evaluated the antimicrobial susceptibility profile and the presence of genes associated with resistance to erythromycin (msrC and ermB) and tetracycline [tet(M) and/or tet(L)] in enterococci isolated from raw sheep's milk and cheeses (colonial, feta-, and pecorino-type) from South region of Brazil. A total of 156 enterococci were isolated from milk (n=80) and cheese (n=76) samples, identified by MALDI-TOF. Enterococcus faecalis (50.6%; n=79) was the most frequent species isolated from both samples. According to in vitro susceptibility tests, enterococci strains were not susceptible to the most commonly antimicrobial agents used in human and veterinary medicine. The frequency of MDR strains in enterococci isolated from milk (53.7%) was higher than those from cheese (24.2%). The tet(M) gene was the most commonly detected among tetracycline not-susceptible strains. The present study provided the first evidence of antimicrobial not-susceptible enterococci in raw sheep's milk and cheeses in South Brazil. Drug-resistant strains, particularly those that are MDR, constitute a One Health issue.

Os enterococos têm sido usados como organismos sentinela para monitorar o padrão de suscetibilidade a antimicrobianos em alimentos, humanos e outros animais. Neste sentido, o presente estudo objetivou avaliar o perfil de susceptibilidade a antimicrobianos e os genes associados com a resistência a eritromicina (msrC and ermB) e à tetraciclina [tet(M) and/or tet(L)] em enterococos isolados de leite cru de ovelha e queijos (colonial, tipo-feta e tipo-pecorino) do Sul do Brasil. Um total de 156 enterococos foram isolados de leite (n=80) e queijo (n=76), identificados por MALDI-TOF. Enterococcus faecalis (50,6%; n=79) foi a espécie mais frequentemente isolada de ambas as amostras. De acordo com o teste de suscetibilidade in vitro, as cepas de enterococos não foram susceptíveis aos agentes antimicrobianos mais comumente utilizados na clínica humana e veterinária. A frequência de cepas de enterococos MDR isoladas do leite (53,7%) foi superior à do queijo (24,2%). O gene tet(M) foi o mais comumente detectado entre as cepas não susceptíveis à tetraciclina. O presente estudo fornece as primeiras evidências de enterococos não susceptíveis aos antimicrobianos em leite cru de ovelha e queijos no Sul do Brasil. Cepas resistentes a drogas, particularmente as que são MDR, representam uma preocupação de Saúde Única.

Metabolic profile, antimicrobial and toxicity evaluation of Azadirachta indica roots / Perfil metabólico, atividade antimicrobiana e de toxicidade das raízes de Azadirachta indica

The constituents of the hydroethanolic extract ofAzadiractaindicaroot were investigated using ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-QTOOF-MSE). Acute toxicity was evaluated in an experimental animal model. We investigated the antibacterial activities ofA. indicaroots againstSalmonella typhimuriumandStaphylococcus aureusand the antifungal activities against strains ofTrichophyton rubrum, Candida albicansandCandida tropicalis. We identified nine secondary metabolites in the hydroethanolic extract by UPLC-QTOOF-MSE. The extract was highly effective in inhibiting the growth of T. rubrum strains, so it can be effective in combating the dermatophyte tested,but it had no inhibition potential on any bacterial strains orCandidaspecies evaluated. It was possible to infer that the extract had no acute toxicity in relation to the animal model Danio rerio. Therefore, since neem has a high bioactive potential and adapts well to the climate of semiarid regions, growing this species could become a source of income for farmers by its use to produce naturals fungicide and drug, as alternatives to conventional products, which can cause microbiological resistance and/or are toxic to the environment, besides being expensive.

Os constituintes do extrato hidroetanólico da raiz deA. indicaforam investigados por cromatografia líquida de ultra-alta performance acoplada à espectrometria de massas do tipo quadrupolo-tempo de voo (UPLC-QTOOF-MSE). A toxicidade aguda foi avaliada em modelo animal. Investigamos as atividades antibacterianas contra Salmonella typhimuriumeStaphylococcus aureuse as atividades antifúngicas contra cepas deTrichophyton rubrum, Candida albicanseCandida tropicalis. Identificamos nove metabólitos secundários no extrato etanólico por UPLC-QTOOF-MSE. O extrato foi altamente eficaz na inibição do crescimento de cepas de T. rubrum, podendo ser eficaz no combate ao dermatofito avaliado, mas não apresentou potencial de inibição em nenhuma cepa bacteriana ou espécies deCandidaavaliadas. Também foi possível inferir que o extrato não apresentou toxicidade aguda em relação ao modelo animalDanio rerio. Portanto, como o Neem tem alto potencial bioativo e se adapta bem ao clima das regiões semiáridas, o cultivo dessa espécie pode se tornar uma fonte de renda para os agricultores a partir da utilização da planta para produção de fungicida e/ou fármaco naturais como alternativa aos produtos convencionais, que podem causar resistência microbiológica e/ou são tóxicas ao meio ambiente, além de serem caros.

Eugenia uniflora: a promising natural alternative against multidrug-resistant bacteria / Eugenia uniflora: uma alternativa natural promissora contra bactérias multirresistentes

This work aimed to evaluate the chemical composition, antioxidant and antimicrobial activities from crude extract and fractions from leaves of Eugenia uniflora Linn. The crude extract was obtained by turbo extraction and their fractions by partitioning. Chromatographic analysis were performed, and the antioxidant capacity was verified by two methods (DPPH• and ABTS•+). The Minimal Inhibitory/Bactericidal Concentration were conducted against twenty-two bacteria, selecting five strains susceptible to extract/fractions and resistant to the antibiotics tested. Ampicillin, azithromycin, ciprofloxacin, and gentamicin were associated with Ethyl Acetate Fraction (EAF) against multidrug-resistant strains in modulatory and checkerboard tests. The chromatographic data showed gallic acid, ellagic acid, and myricitrin in crude extract, with enrichment in the EAF. The electron transfer activity demonstrated in the antioxidant tests is related to the presence of flavonoids. The Gram-positive strains were more susceptible to EAF, and their action spectra were improved by association, comprising Gram-negative bacilli. Synergisms were observed to ciprofloxacin and gentamicin against Pseudomonas aeruginosa colistin-resistant. The results demonstrate that the extract and enriched fraction obtained from the leaves of E. uniflora act as a promising natural alternative against multidrug-resistant bacteria.

Este trabalho teve como objetivo avaliar a composição química, as atividades antioxidantes e antimicrobianas do extrato bruto e frações de folhas de Eugenia uniflora Linn. O extrato bruto foi obtido por turbólise e suas frações por partição. Foram realizadas análises cromatográficas e a capacidade antioxidante foi verificada por dois métodos (DPPH• e ABTS•+). A Concentração Inibitória/Bactericida Mínima foi realizada contra vinte e duas bactérias, selecionando cinco cepas suscetíveis a extração/frações e resistentes aos antibióticos testados. Ampicilina, azitromicina, ciprofloxacina e gentamicina foram associados à Fração Acetato de Etila (FAE) contra cepas multirresistentes em testes modulatórios e de checkboard. Os dados cromatográficos mostraram ácido gálico, ácido elágico e miricitrina em extrato bruto, com enriquecimento na FAE. A atividade de transferência de elétrons demonstrada nos testes antioxidantes está relacionada com a presença de flavonoides. As cepas de Gram-positivas foram mais suscetíveis à FAE, e seus espectros de ação foram melhorados por associação, compreendendo bacilos Gram-negativos. Foram observados sinergismos de ciprofloxacina e gentamicina contra Pseudomonas aeruginosa resistente à colistina. Os resultados demonstram que o extrato e a fração enriquecida obtida das folhas de E. uniflora atuam como uma alternativa natural promissora contra bactérias multirresistentes.

Essential oils isolated from popular medicinal plants and spices as alternative antimicrobial and antibiofilm compounds against the pig pathogen Actinobacilluspleuropneumoniae / Óleos essenciais isolados de plantas medicinais e temperos populares como compostos antimicrobianos e antibiofilme alternativos contra o patógeno de suínosActinobacillus pleuropneumoniae

Actinobacilluspleuropneumoniae is the causative agent of swine pleuropneumonia, a contagious respiratory disease associated with high morbidity and economic losses. While antibiotic therapy helps to control the spreading of the pathogen on the farm, resistance to several classes of antibiotics were reported, and treatment can be impaired by the bacterial ability to form biofilms. This increases the need for alternative therapy approaches, including the use of natural compounds with antimicrobial and/or antibiofilm activities. In this research we analyzed, by the broth microdilution method, the inhibitory and bactericidal activities of the essential oils obtained from eighteen Brazilian popular medicinal plants or spices against clinical isolates of Actinobacilluspleuropneumoniae. After that, sub-inhibitory concentrations of active oils were tested for their antibiofilm effects, analyzed by the crystal violet method. Among the eighteen oils tested, eight (extracted from cinnamon, coriander, peppermint, spearmint, thyme, marjoram, eucalyptus, and laurel) presented bacteriostatic and bactericidal activities against all isolates, and subinhibitory concentrations of five of them disrupted up to 80% preformed biofilms, and significantly inhibited biofilm formation. The chemical composition of such oils was assessed by gas chromatography/mass spectrometry (GC/MS) and is presented, indicating that their bactericidal and antibiofilm properties were mostly associated with the presence of monoterpenes and phenylpropanoids. To our knowledge, this is the first report of essential oils with potential to control environmental contamination and animal infection with A. pleuropneumoniae, representing an alternative to increasing levels of antibiotic resistance.

Actinobacillus pleuropneumoniae é o agente causador da pleuropneumonia suína, uma doença respiratória contagiosa associada à alta morbidade e perdas econômicas. Embora a antibioticoterapia ajude a controlar a disseminação do patógeno na fazenda, a resistência a várias classes de antibióticos tem sido relatada e o tratamento pode ser prejudicado pela capacidade bacteriana de formar biofilmes. Isso aumenta a necessidade de abordagens terapêuticas alternativas, que incluem o uso de compostos naturais com atividades antimicrobianas e/ou antibiofilme. Neste trabalho, analisamos, pelo método de microdiluição em caldo nutriente, os efeitos inibitórios e bactericidas dos óleos essenciais obtidos de dezoito plantas medicinais brasileiras populares e/ou temperos contra isolados clínicos de Actinobacillus pleuropneumoniae. Então, concentrações subinibitórias dos óleos ativos foram testados quanto a suas atividades antibiofilme, analisados pelo método do cristal violeta. Dentre os dezoito óleos testados, oito (extraídos da canela, coentro, hortelã-pimenta, hortelã, tomilho, manjerona, eucalipto e louro) apresentaram atividade bacteriostática e bactericida contra todos os isolados, e concentrações subinibitórias de cinco deles romperam biofilmes pré-formados em até 80%, além de inibirem fortemente a formação de biofilmes. A composição química desses óleos foi avaliada por cromatografia gasosa/espectrometria de massa (CG/MS) e é apresentada, indicando que suas propriedades bactericidas e antibiofilme estavam principalmente associadas à presença de monoterpenos e fenilpropanóides. Este é o primeiro relato de óleos essenciais com potencial para controlar a contaminação ambiental e infecção animal por A. pleuropneumoniae, representando uma alternativa contra o crescente aumento de resistência bacteriana aos antibióticos.

Metabolic profile, antimicrobial and toxicity evaluation of Azadirachta indica roots / Perfil metabólico, atividade antimicrobiana e de toxicidade das raízes de Azadirachta indica

ABSTRACT: The constituents of the hydroethanolic extract ofAzadiractaindicaroot were investigated using ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-QTOOF-MSE). Acute toxicity was evaluated in an experimental animal model. We investigated the antibacterial activities ofA. indicaroots againstSalmonella typhimuriumandStaphylococcus aureusand the antifungal activities against strains ofTrichophyton rubrum, Candida albicansandCandida tropicalis. We identified nine secondary metabolites in the hydroethanolic extract by UPLC-QTOOF-MSE. The extract was highly effective in inhibiting the growth of T. rubrum strains, so it can be effective in combating the dermatophyte tested,but it had no inhibition potential on any bacterial strains orCandidaspecies evaluated. It was possible to infer that the extract had no acute toxicity in relation to the animal model Danio rerio. Therefore, since neem has a high bioactive potential and adapts well to the climate of semiarid regions, growing this species could become a source of income for farmers by its use to produce naturals fungicide and drug, as alternatives to conventional products, which can cause microbiological resistance and/or are toxic to the environment, besides being expensive.

RESUMO: Os constituintes do extrato hidroetanólico da raiz deA. indicaforam investigados por cromatografia líquida de ultra-alta performance acoplada à espectrometria de massas do tipo quadrupolo-tempo de voo (UPLC-QTOOF-MSE). A toxicidade aguda foi avaliada em modelo animal. Investigamos as atividades antibacterianas contra Salmonella typhimuriumeStaphylococcus aureuse as atividades antifúngicas contra cepas deTrichophyton rubrum, Candida albicanseCandida tropicalis. Identificamos nove metabólitos secundários no extrato etanólico por UPLC-QTOOF-MSE. O extrato foi altamente eficaz na inibição do crescimento de cepas de T. rubrum, podendo ser eficaz no combate ao dermatofito avaliado, mas não apresentou potencial de inibição em nenhuma cepa bacteriana ou espécies deCandidaavaliadas. Também foi possível inferir que o extrato não apresentou toxicidade aguda em relação ao modelo animalDanio rerio. Portanto, como o Neem tem alto potencial bioativo e se adapta bem ao clima das regiões semiáridas, o cultivo dessa espécie pode se tornar uma fonte de renda para os agricultores a partir da utilização da planta para produção de fungicida e/ou fármaco naturais como alternativa aos produtos convencionais, que podem causar resistência microbiológica e/ou são tóxicas ao meio ambiente, além de serem caros.

Temporal association between antibiotic use and resistance in Gram-negative bacteria / Associação temporal entre o uso de antibióticos e resistência em bactérias gram-negativas

Abstract The β-lactam/lactamase inhibitors (BLBLIs) combination drugs are considered an effective alternative to carbapenems. However, there is a growing concern that the increased use of BLBLIs may lead to increased resistance. This study determined the temporal association between the consumption of BLBLI and the antimicrobial resistance in Gram-negative bacteria. In this retrospective study, electronic data on the Gram-negative bacterial isolates, including A. baumannii, P. aeruginosa, E. coli, and K. pneumoniae from in-patients and susceptibility testing results were retrieved from the medical records of the clinical laboratory. A linear regression and cross-correlation analysis were performed on the acquired data. Increasing trends (p<0.05) in the consumption of BIBLI and carbapenem with a median use of 27.68 and 34.46 DDD/1000 PD per quarter were observed, respectively. A decreased trend (p=0.023) in the consumption of fluoroquinolones with a median use of 29.13 DDD/1000 PD per quarter was observed. The resistance rate of K. pneumoniae was synchronized with the BIBLI and carbapenem consumptions with a correlation coefficient of 0.893 (p=0.012) and 0.951 (p=0.016), respectively. The cross-correlation analysis against the consumption of BIBLI and meropenem resistant K. pneumoniae was peaked at 0-quarter lag (r=951, p=0.016). There was an increasing trend in the consumption of BLBLI and carbapenems. The increasing trend in the rates of resistance to piperacillin/tazobactam, in line with the increasing consumption of BLBLI, suggests that BLBLI has to be used with caution and cannot be directly considered as a long-term alternative to carbapenems.

Resumo Os medicamentos combinados de β-lactâmicos / inibidores da lactamase (BLBLIs) são considerados uma alternativa eficaz aos carbapenêmicos. No entanto, existe uma preocupação crescente de que o aumento do uso de BLBLIs pode levar ao aumento da resistência. Este estudo determinou a associação temporal entre o consumo de BLBLI e a resistência antimicrobiana em bactérias gram-negativas. Neste estudo retrospectivo, os dados eletrônicos sobre as bactérias gram-negativas isoladas, incluindo A. baumannii, P. aeruginosa, E. coli e K. pneumoniae de pacientes internados e os resultados dos testes de suscetibilidade foram recuperados dos registros médicos do laboratório clínico. Uma regressão linear e análise de correlação cruzada foram realizadas nos dados adquiridos. Foram observadas tendências crescentes (p < 0,05) no consumo de BIBLI e carbapenem com uma mediana de uso de 27,68 e 34,46 DDD/1000 PD por trimestre, respectivamente. Foi observada uma tendência de diminuição (p = 0,023) no consumo de fluoroquinolonas com uma mediana de uso de 29,13 DDD/1000 PD por trimestre. A taxa de resistência de K. pneumoniae foi sincronizada com os consumos de BIBLI e carbapenem com coeficiente de correlação de 0,893 (p = 0,012) e 0,951 (p = 0,016), respectivamente. A análise de correlação cruzada contra o consumo de BIBLI e K. pneumoniae resistente ao meropenem atingiu o pico no intervalo de 0 quarto (r = 951, p = 0,016). Houve uma tendência de aumento no consumo de BLBLI e carbapenêmicos. A tendência crescente nas taxas de resistência a piperacilina/tazobactam, em linha com o consumo crescente de BLBLI, sugere que BLBLI deve ser usado com cautela e não pode ser considerado diretamente como alternativa de longo prazo aos carbapenêmicos.

A comparative study to evaluate the effects of antibiotics, plant extracts and fluoride-based toothpaste on the oral pathogens isolated from patients with gum diseases in Pakistan / Um estudo comparativo para avaliar os efeitos de antibióticos, extratos vegetais e creme dental à base de flúor sobre os patógenos orais isolados de pacientes com doenças gengivais no Paquistão

Abstract Oral diseases caused by various microorganisms are common around the world. Scientific research has now been focusing on novel medicines to overcome bacterial resistance and antibiotics side effects; therefore, the current study was designed to assess the efficacy of certain antibiotics, toothpaste, and medicinal plant extracts (Ajuga bracteosa and Curcuma longa) versus the bacterial pathogens isolated from the human oral cavity. A total of 130 samples were collected from Khyber Teaching Hospital Peshawar, Pakistan, among those 27 species isolated, and eight bacterial species were identified from the samples. Among all the bacterial species, Staphylococcus aureus (29.62%) and Proteus mirabilis (22.2%) were found to be more prevalent oral pathogens. In comparison, the least pervasive microbes were Proteus vulgaris, Shigella sonnei, Escherichia coli and Aeromonas hydrophila. The study also suggested that dental problems were more prevalent in males (41-50 years of age) than females. Among the eight antibiotics used in the study, the most promising results were shown by Foxicillin against A. hydrophila. The survey of TP1 revealed that it showed more potent antagonist activity against Proteus vulgaris as compared TP2 and TP3 that might be due to the high content of fluoride. The Curcuma longa showed more significant activity than Ajuga bracteosa (Stem, leaves and root) extracts. The data obtained through this study revealed that antibiotics were more effective for oral bacterial pathogens than toothpaste and plant extracts which showed moderate and low activity, respectively. Therefore, it is suggested that the active compounds in individual medicinal plants like Curcuma longa and Ajuga bracteosa could replace the antibiotics when used in daily routine as tooth cleansers or mouth rinses.

Resumo As doenças bucais causadas por vários microrganismos são comuns em todo o mundo. A pesquisa científica agora tem se concentrado em novos medicamentos para superar a resistência bacteriana e os efeitos colaterais dos antibióticos; portanto, o presente estudo foi desenhado para avaliar a eficácia de certos antibióticos, pasta de dente e extratos de plantas medicinais (Ajuga bracteosa e Curcuma longa) contra os patógenos bacterianos isolados da cavidade oral humana. No total, 130 amostras foram coletadas do Khyber Teaching Hospital Peshawar, Paquistão, entre essas, 27 espécies foram isoladas e oito espécies bacterianas foram identificadas a partir das amostras. Entre todas as espécies bacterianas, Staphylococcus aureus (29.62%) e Proteus mirabilis (22.2%) foram os patógenos orais mais prevalentes. Em comparação, os micróbios menos difundidos foram Proteus vulgaris, Shigella sonnei, Escherichia coli e Aeromonas hydrophila. O estudo também sugeriu que os problemas dentários eram mais prevalentes em homens (41-50 anos de idade) do que em mulheres. Entre os oito antibióticos usados ​​no estudo, os resultados mais promissores foram mostrados pelo Foxicillin contra A. hydrophila. A pesquisa de TP1 revelou que ele mostrou atividade antagonista mais potente contra Proteus vulgaris em comparação a TP2 e TP3, o que pode ser devido ao alto teor de flúor. A Curcuma longa apresentou atividade mais significativa em relação aos extratos de Ajuga bracteosa (caule, folhas e raiz). Os dados obtidos neste estudo revelaram que os antibióticos foram mais eficazes para os patógenos bacterianos orais do que os dentifrícios e os extratos vegetais que apresentaram atividade moderada e baixa, respectivamente. Portanto, sugere-se que os compostos ativos em plantas medicinais individuais como Curcuma longa e Ajuga bracteosa possam substituir os antibióticos quando usados ​​na rotina diária como limpadores de dentes ou enxaguatórios bucais.

Portrayal of Punica granatum L peel extract through High Performance Liquid Chromatography and antimicrobial activity evaluation / Retrato do extrato de casca de Punica granatum L por meio de cromatografia líquida de alto desempenho e avaliação da atividade antimicrobiana

ncreasing trend in antimicrobial resistance and failure of chemically synthesized antibiotics lead to discover alternative methods for the treatment of bacterial infections. Various medicinal plants are in use traditionally and their active compounds can be further applied for treatment of bacterial diseases. This study was designed to determine the antibacterial activity of Punica granatum (P. granatum L.) (pomegranate) peel extract against Enterobacteriaceae [Escherichia coli (E. coli), Salmonella Typhimurium (S. Typhimurium) and Shigella Dysenteriae (S. Dysenteriae)] and gram-positive bacterium [Staphylococcus aureus (Staph aureus)]. Methanolic extract of P. granatum L. peel was prepared by Soxhlet apparatus method. Total flavonoid and phenolic contents from the extract were determined by High Performance Liquid Chromatography (HPLC). The antibacterial activity of P. granatum L. peel extract was evaluated through agar well diffusion method. HPLC showed the range of phenolics (gallic acid, caffeic acid, benzoic acid, cinnamic acid) and flavonoid compounds. The chemical structures of flavonoid and phenolics found in the methanolic extract of P. granatum L. peel have been reported for the first time. The methanolic peel extract (50 ul) of yellow P. granatum L. showed 26, 10, 10 and 9mm zones of inhibition (ZOI) against S. aureus, S. Typhimurium, S. Dysenteriae and E. coli, respectively. The methanolic extract of red P. granatum L. (100 ul) showed 27, 8, 12 and 15 mm ZOI against Staph. aureus, S. Typhimurium, S. Dysenteriae and E. coli, respectively. Highest ZOI was observed against Staph. aureus. Many of the bacteria studied in the present work may cause serious gastrointestinal infections, which can lead to hemorrhagic diarrhea in children. These [...].

A tendência crescente na resistência antimicrobiana e na falha dos antibióticos sintetizados quimicamente leva à descoberta de métodos alternativos para o tratamento de infecções bacterianas. Várias plantas medicinais estão em uso tradicionalmente e seus compostos ativos podem ser posteriormente aplicados para o tratamento de doenças bacterianas. Este estudo foi desenhado para determinar a atividade antibacteriana do extrato de casca de Punica granatum (P. granatum L.) (romã) contra Enterobacteriaceae [Escherichia coli (E. coli), Salmonella Typhimurium (S. Typhimurium) e Shigella Dysenteriae (S. Dysenteriae) ] e bactéria gram-positiva [Staphylococcus aureus (Staph aureus)]. O extrato metanólico da casca de P. granatum L. foi preparado pelo método do aparelho de Soxhlet. O conteúdo total de flavonoides e fenólicos do extrato foi determinado por cromatografia líquida de alta eficiência (HPLC). A atividade antibacteriana do extrato da casca de P. granatum L. foi avaliada através do método de difusão em ágar. HPLC mostrou a gama de compostos fenólicos (ácido gálico, ácido cafeico, ácido benzoico, ácido cinâmico) e flavonoides. As estruturas químicas de flavonoides e fenólicos encontradas no extrato metanólico da casca de P. granatum L. foram relatadas pela primeira vez. O extrato metanólico da casca (50 ul) de P. granatum L. amarelo apresentou zonas de inibição (ZOI) de 26, 10, 10 e 9mm contra S. aureus, S. Typhimurium, S. Dysenteriae e E. coli, respectivamente. O extrato metanólico de P. granatum L. vermelho (100 ul) apresentou 27, 8, 12 e 15 mm IOI contra Staph. aureus, S. Typhimurium, S. Dysenteriae e E. coli, respectivamente. O ZOI mais alto foi observado contra Staph. aureus. Muitas das bactérias estudadas no presente trabalho podem causar infecções gastrointestinais graves, que podem levar à diarreia [...].

Portrayal of Punica granatum L peel extract through High Performance Liquid Chromatography and antimicrobial activity evaluation / Retrato do extrato de casca de Punica granatum L por meio de cromatografia líquida de alto desempenho e avaliação da atividade antimicrobiana

ncreasing trend in antimicrobial resistance and failure of chemically synthesized antibiotics lead to discover alternative methods for the treatment of bacterial infections. Various medicinal plants are in use traditionally and their active compounds can be further applied for treatment of bacterial diseases. This study was designed to determine the antibacterial activity of Punica granatum (P. granatum L.) (pomegranate) peel extract against Enterobacteriaceae [Escherichia coli (E. coli), Salmonella Typhimurium (S. Typhimurium) and Shigella Dysenteriae (S. Dysenteriae)] and gram-positive bacterium [Staphylococcus aureus (Staph aureus)]. Methanolic extract of P. granatum L. peel was prepared by Soxhlet apparatus method. Total flavonoid and phenolic contents from the extract were determined by High Performance Liquid Chromatography (HPLC). The antibacterial activity of P. granatum L. peel extract was evaluated through agar well diffusion method. HPLC showed the range of phenolics (gallic acid, caffeic acid, benzoic acid, cinnamic acid) and flavonoid compounds. The chemical structures of flavonoid and phenolics found in the methanolic extract of P. granatum L. peel have been reported for the first time. The methanolic peel extract (50 ul) of yellow P. granatum L. showed 26, 10, 10 and 9mm zones of inhibition (ZOI) against S. aureus, S. Typhimurium, S. Dysenteriae and E. coli, respectively. The methanolic extract of red P. granatum L. (100 ul) showed 27, 8, 12 and 15 mm ZOI against Staph. aureus, S. Typhimurium, S. Dysenteriae and E. coli, respectively. Highest ZOI was observed against Staph. aureus. Many of the bacteria studied in the present work may cause serious gastrointestinal infections, which can lead to hemorrhagic diarrhea in children. These [...].(AU)

A tendência crescente na resistência antimicrobiana e na falha dos antibióticos sintetizados quimicamente leva à descoberta de métodos alternativos para o tratamento de infecções bacterianas. Várias plantas medicinais estão em uso tradicionalmente e seus compostos ativos podem ser posteriormente aplicados para o tratamento de doenças bacterianas. Este estudo foi desenhado para determinar a atividade antibacteriana do extrato de casca de Punica granatum (P. granatum L.) (romã) contra Enterobacteriaceae [Escherichia coli (E. coli), Salmonella Typhimurium (S. Typhimurium) e Shigella Dysenteriae (S. Dysenteriae) ] e bactéria gram-positiva [Staphylococcus aureus (Staph aureus)]. O extrato metanólico da casca de P. granatum L. foi preparado pelo método do aparelho de Soxhlet. O conteúdo total de flavonoides e fenólicos do extrato foi determinado por cromatografia líquida de alta eficiência (HPLC). A atividade antibacteriana do extrato da casca de P. granatum L. foi avaliada através do método de difusão em ágar. HPLC mostrou a gama de compostos fenólicos (ácido gálico, ácido cafeico, ácido benzoico, ácido cinâmico) e flavonoides. As estruturas químicas de flavonoides e fenólicos encontradas no extrato metanólico da casca de P. granatum L. foram relatadas pela primeira vez. O extrato metanólico da casca (50 ul) de P. granatum L. amarelo apresentou zonas de inibição (ZOI) de 26, 10, 10 e 9mm contra S. aureus, S. Typhimurium, S. Dysenteriae e E. coli, respectivamente. O extrato metanólico de P. granatum L. vermelho (100 ul) apresentou 27, 8, 12 e 15 mm IOI contra Staph. aureus, S. Typhimurium, S. Dysenteriae e E. coli, respectivamente. O ZOI mais alto foi observado contra Staph. aureus. Muitas das bactérias estudadas no presente trabalho podem causar infecções gastrointestinais graves, que podem levar à diarreia [...].(AU)

Detection of microorganisms with antibacterial activities from different industrial wastes and GC-MS analysis of crude microbial extract / Detecção de microrganismos com atividades antibacterianas de diferentes resíduos industriais e análise por GC-MS do extrato microbiano bruto

Abstract Many soil microorganisms' i.e., bacteria and fungi produce secondary metabolites called antibiotics. These are used for the treatment of some of the bacterial, fungal and protozoal diseases of humans. There is a need for isolation of a broad spectrum of antibiotics from microorganisms due to the emergence of antibiotic resistance. In the present study two antibiotic producing bacteria Klebsiella pneumoniae and Bacillus cereus were isolated from pharmaceutical and poultry feed industry of Hattar, Haripur Pakistan. Total 10 waste samples were collected from different industries (Marble, Ghee, Soap, Mineral, Steel, Poultry Feed, Pharmaceutical, Qarshi, Cosmetic and Glass). Thirty-three bacterial strains were isolated from industrial wastes of these ten different industries. Fourteen out of thirty-three bacterial strains exhibited antimicrobial activities against at least one of the test microbes considered in this study including Escherchia coli, Staphylococcus aureus and Salmonella typhi. The bacteria were isolated by standard serial dilution spread plate technique. Morphological characterization of the isolates was done by Gram staining. Nine bacterial isolates out of fourteen were initially identified as B. cereus and five as K. pneumoniae through biochemical characterization. The antibacterial activities were tested by well diffusion method. Maximum number of antibiotic producing bacteria were isolated from pharmaceutical and poultry feed industry based on the results of primary screening, the most potential isolates S9, S19, S20, S22 and S23 were selected for secondary screening. The maximum activity against E. coli and S. aureus was recorded by bacterial isolate S19 i.e zones of inhibition of 6.5mm and 9mm while S20 showed 7.5mm and 6mm zones respectively. Molecular identification was carried out on the basis of 16S rRNA sequence analysis. Finally, the isolates were identified as B. cereus accession number LC538271and K. pneumoniae accession number MT078679. Analysis of bacterial extract S20 through GC-MS indicated the presence of 8 compounds of diverse nature and structure. Present study suggests that wastes of pharmaceutical and poultry feed industry may have antibiotic producing bacteria. These bacteria could be utilized for the production of antibiotics. B. cereus and K. pneumoniae isolated from wastes of poultry feed and pharmaceutical industries have the potential to produce antibiotics and could be used to control the microbial growth.

Resumo Muitos microrganismos do solo, ou seja, bactérias e fungos produzem metabólitos secundários chamados antibióticos. Eles são usados ​​para tratamento de algumas doenças bacterianas, fúngicas e protozoárias em humanos. Há necessidade de isolamento de um amplo espectro de antibióticos de microrganismos devido ao surgimento de resistência aos antibióticos. No presente estudo, duas bactérias produtoras de antibióticos, Klebsiella pneumoniae e Bacillus cereus, foram isoladas da indústria farmacêutica e de ração avícola de Hattar, Haripur, Paquistão. Um total de 10 amostras de resíduos foi coletado de diferentes indústrias (mármore, ghee, sabão, mineral, aço, ração para aves, farmacêutica, Qarshi, cosmética e vidro). Trinta e três cepas bacterianas foram isoladas de resíduos industriais dessas dez diferentes indústrias. Quatorze das 33 cepas bacterianas exibiram atividades antimicrobianas contra pelo menos um dos micróbios de teste considerados neste estudo, incluindo Escherchia coli, Staphylococcus aureus e Salmonella typhi. As bactérias foram isoladas pela técnica de placa de diluição em série padrão. A caracterização morfológica dos isolados foi feita por coloração de gram. Nove isolados bacterianos de 14 foram inicialmente identificados como B. cereus e cinco como K. pneumoniae por meio de caracterização bioquímica. As atividades antibacterianas foram testadas pelo método de difusão em poço. O número máximo de bactérias produtoras de antibióticos foi isolado da indústria farmacêutica e de ração avícola com base nos resultados da triagem primária, os isolados mais potenciais S9, S19, S20, S22 e S23 foram selecionados para a triagem secundária. A atividade máxima contra E. coli e S. aureus foi registrada pelo isolado bacteriano S19, ou seja, zonas de inibição de 6,5 mm e 9 mm, enquanto S20 mostrou zonas de 7,5 mm e 6 mm, respectivamente. A identificação molecular foi realizada com base na análise da sequência 16S rRNA. Finalmente, os isolados foram identificados como B. cereus número de acesso LC538271 e K. pneumoniae número de acesso MT078679. A análise do extrato bacteriano S20 por meio de GC-MS indicou a presença de oito compostos de natureza e estrutura diversas. O presente estudo sugere que resíduos da indústria farmacêutica e de ração para aves podem conter bactérias produtoras de antibióticos. Essas bactérias podem ser utilizadas para a produção de antibióticos B. cereus e K. pneumoniae isolados de resíduos de rações de aves e indústrias farmacêuticas têm potencial para produzir antibióticos e podem ser usados ​​para controlar o crescimento microbiano.

Profile of external otitis isolates in dogs and cats diagnosed in laboratory of microbiology in 2020 / Profile of otitis externa isolates in dogs and cats diagnosed in laboratory of microbiology in 2020

Otitis externa are one of the most frequent diseases in the clinical routine of dogs and cats and they can cause by several associated factors. Due to incorrect use of antimicrobial, the treatment and the control of otitis have become challenging. The objective of this study is analyze results of otological examinations of the Laboratory of Microbiology in 2020, and to demonstrate the profile of patients and isolated bacteria. Staphylococcus was the main genus isolated and 71,11% of samples showed multi drug resistance to antimicrobial tested. This results show the need to use of complementary exams to control of otitis externa. Keywords: canines, felines, otological diseases, antimicrobial susceptibilities.

Otitis externa is one of the most frequent diseases in the clinicalroutine of dogs and cats, and they can be caused by several associated factors.Due to incorrect use of antimicrobial products, the treatment and control of otitishave become challenging. This study aims to analyze the results of otologicalexams at the Laboratory of Microbiology HV-ULBRA in 2020 and demonstratethe profie of patients and isolated bacteria. Staphylococcus was the main genusisolated, and 71,11% of samples showed multi-drug resistance to antimicrobialtesting. These results indicate the need to use complementary examinations tocontrol otitis externa.  Keywords: canines, felines, otological diseases, antimicrobial susceptibility test

Isolation and antimicrobial resistance of coagulase-negative staphylococci recovered from healthy tortoises in Minas Gerais, Brazil / Isolamento e resistência aos antimicrobianos de estafilococos coagulase negativos recuperados de jabutis saudáveis em Minas Gerais, Brasil

In the last few decades, there has been a global increase in the adoption of reptiles as companion animals, mainly turtles and tortoises. Considering the popularity of reptiles as pets in Brazil, and a notable lack of data about potentially pathogenic staphylococci in these animals, this study isolated and evaluate the antimicrobial susceptibility of staphylococcal species from healthy tortoises (Chelonoidis carbonaria) in Brazil. During a 12-month period (February 2019 to February 2020), cloacal swabs from 66 healthy tortoises were collected at the Wild Animals Screening Center in Belo Horizonte, Minas Gerais, Brazil. The swabs were plated onto mannitol salt agar for staphylococci isolation, and species identification was performed using MALDI-TOF MS. Antimicrobial susceptibility was investigated using the disk diffusion method, and the presence of the mecA gene was investigated by PCR to detect methicillin resistance. Of the tested animals, 72.7% were positive for staphylococcal isolation. All isolates were coagulase-negative staphylococci (CoNS), and Staphylococcus sciuri (81.3%), and S. xylosus (12.5%) were the most frequently isolated species. The majority of the isolates (56%) were resistant to at least one antimicrobial agent. A high frequency of resistance was observed for penicillin (35.5%) and tetracycline (29.1 %). All strains were susceptible to cefoxitin, chloramphenicol, ciprofloxacin, erythromycin, and gentamicin. All isolates were negative for the mecA gene. The present work suggests that healthy tortoises are mainly colonized by CoNS, especially S. sciuri. Half of the isolates were resistant to at least one antimicrobial, raising questions regarding the possible role of these animals as reservoirs of antimicrobial resistance genes.

Nas últimas décadas, houve um aumento global na adoção de répteis como animais de companhia, principalmente tartarugas e jabutis. Considerando a popularidade dos répteis como animais de estimação no Brasil e a notável falta de dados sobre estafilococos potencialmente patogênicos nesses animais, o objetivo deste estudo foi isolar e avaliar a susceptibilidade antimicrobiana de espécies estafilocócicas de jabutis (Chelonoidis carbonaria) saudáveis no Brasil. Durante um período de 12 meses (fevereiro de 2019 a fevereiro de 2020), suabes cloacais de 66 jabutis saudáveis foram coletados no Centro de Triagem de Animais Silvestres em Belo Horizonte, Minas Gerais, Brasil. Os suabes foram plaqueados em ágar manitol salgado para isolamento de estafilococos e a identificação das espécies foi realizada usando MALDI-TOF MS. A susceptibilidade aos antimicrobianos foi investigada pelo método de difusão em disco, e a presença do gene mecA foi investigada por PCR para detectar resistência à meticilina. Dos animais testados, 72,7% foram positivos para o isolamento estafilocócico. Todos os isolados eram estafilococos coagulase-negativos (CoNS), sendo Staphylococcus sciuri (81,3%) e S. xylosus (12,5%) as espécies mais frequentemente isoladas. A maioria dos isolados (56%) foi resistente a pelo menos um antimicrobiano. Alta frequência de resistência foi observada para penicilina (35,5%) e tetraciclina (29,1%). Todas as estirpes foram sensíveis à cefoxitina, cloranfenicol, ciprofloxacina, eritromicina e gentamicina. Todos os isolados foram negativos para o gene mecA. O presente trabalho sugere que jabutis saudáveis são colonizados principalmente por CoNS, especialmente S. sciuri. Metade dos isolados foram resistentes a pelo menos um antimicrobiano, levantando questões sobre o possível papel desses animais como reservatórios de genes de resistência aos antimicrobianos.

Canine cystitis - biofilm formation by bacterial isolates

Background: Biofilms have been reported as important virulent markers associated with drug resistance in urinary tractinfections (UTIs) in humans and dogs. However, in veterinary medicine, researches involving biofilm formation, treatments and preventions have been limited; yet, it is still possible to find few studies demonstrating biofilm-forming bacteriaassociated with different comorbidities such as otitis, wound infections, UTIs, and endometritis. These studies generallyselect dogs with chronic and recurrent infections, which could be an important factor in antibiotic resistance. We aimed toevaluate biofilms in sporadic cystitis regarding prevalence and drug resistance.Materials, Methods & Results: Urine samples were collected by cystocentesis from 36 client-owned dogs under clinicaland laboratory suspicion of non-recurrent urinary bladder infection (cystitis). Urine was aseptically plated onto bloodagar, MacConkey, and CLED, followed by incubation for 24 to 48 h. Definitive identification of a potential pathogen wasmade by subculture collected from an isolated colony to obtain a pure culture. The gram staining method and specificbiochemical tests (phenol red fermentation, lysine, phenylalanine, citrate, sulfide-indole-motility, and urease) were usedto distinguish and classify the bacteria. After identification, the bacteria were tested for antimicrobial susceptibility by astandard disk diffusion method, using the following antimicrobials: amoxicillin with clavulanic acid, ampicillin, ceftriaxone, ciprofloxacin, clindamycin, cefazolin, cephalothin, erythromycin, gentamicin, norfloxacin, and sulfamethoxazoletrimethoprim. The biofilm-forming ability was determined based on a culture...

Avaliação do uso de antimicrobianos e suas interações medicamentosas em cães e gatos hospitalizados / Evaluation of antimicrobial use and their drug interactions in hospitalized dogs and cats

Os antimicrobianos são amplamente utilizados na medicina veterinária, portanto são necessários critérios na hora da prescrição desses fármacos a fim de evitar a resistência bacteriana que se tornou uma grande preocupação tanto para a saúde humana quanto para a medicina veterinária, devido ao seu uso indiscriminado. Uma técnica muito utilizada na rotina, são as associações entre antimicrobianos, que combinam o mecanismo de ação de cada fármaco para se obter os efeitos desejados, acarretando nas interações medicamentosas. Porém, se utilizado de forma incorreta, essas interações podem ser prejudiciais para a saúde dos pacientes. O objetivo desse estudo foi analisar prontuários de pacientes caninos e felinos hospitalizados afim de avaliar os critérios de seleção para o uso de antimicrobianos e suas associações. Foram analisados 112 prontuários, sendo eles 24 felinos e 88 caninos atendidos e hospitalizados no Hospital Veterinário "Prof. Ricardo Alexandre Rippler" da Universidade Vila Velha-UVV. O estudo demonstrou que o metronidazol foi o antimicrobiano mais utilizado em cães e a amoxicilina com clavulanato de potássio foi o mais utilizado nos felinos, além disso, a amoxicilina com clavulanato de potássio e metronidazol foi a associação mais observada em cães e gatos. A maior parte dos pacientes fizeram uso dos antibióticos de forma empírica, o que pode contribuir para a resistência microbiana. Além disso, foram encontradas algumas interações entre antimicrobianos de efeitos antagônicos, porém a maioria das associações foram realizadas de forma correta provocando efeito sinérgico no organismo dos pacientes. Conclui-se que o conhecimento sobre a utilização dos antimicrobianos na rotina da internação de cães e gatos é importante e com base nesses conhecimentos podemos aplicar medidas para atenuar a resistência aos antimicrobianos.

Antimicrobials are widely used in veterinary medicine, so criteria are needed when prescribing these drugs in order to avoid bacterial resistance, which has become a major concern for both human health and veterinary medicine, due to its indiscriminate use. A technique widely used in routine is the association between antimicrobials, which combine the mechanism of action of each drug to obtain the desired effects, resulting in drug interactions. However, if used incorrectly, these interactions can be harmful to the health of patients. The aim of this study was to analyze the medical records of hospitalized canine and feline patients in order to assess the selection criteria for the use of antimicrobials and their associations. 112 medical records were analyzed, of which 24 were cats and 88 canines treated and hospitalized at the "Prof. Ricardo Alexandre Rippler" from Universidade Vila Velha-UVV. The study showed that metronidazole was the most used antimicrobial in dogs and amoxicillin with potassium clavulanate was the most used in felines. Furthermore, amoxicillin with potassium clavolanate and metronidazole was the most observed association in dogs and cats. Most patients used antibiotics empirically, which may contribute to microbial resistance. In addition, some interactions were found between antimicrobials with antagonistic effects, but most associations were correctly performed, causing a synergistic effect in the patients' bodies. It is concluded that knowledge about the use of antimicrobials in the routine hospitalization of dogs and cats is important and based on this knowledge we can apply measures to mitigate antimicrobial resistance.

Antimicrobial resistance profile of bacteria isolated from canine and feline samples at the Preventive Veterinary Laboratory of the Federal University of Rio Grande do Sul (UFRGS) - Brazil

Background: Antimicrobial resistance (AMR) is a major global health threat. In small animals such as dogs and cats, antimicrobials are most commonly prescribed for skin and genitourinary diseases; therefore, the AMR of bacteria involved in these infections should be monitored. In addition, the results of antimicrobial susceptibility testing (AST) may be interpreted as a local epidemiological history of AMR. The Preventive Veterinary Medicine Laboratory (PVML) received clinical samples from dogs and cats for bacterial isolation and AST. Thus, this study aimed to assess the AMR of bacteria isolated from the samples of dogs and cats received at the Preventive Veterinary Medicine Laboratory (PVML). Materials, Methods & Results: Data from bacteriological examinations performed at the PVML of the Universidade Federal do Rio Grande do Sul (UFRGS) during 5 years were analyzed. Skin and ear canal samples were inoculated in 5% sheep blood agar, and urine samples were streaked on CHROMagar™ orientation. After incubation at 36±1°C for up to 72 h, identification and AST were performed according to routine protocols. Of 1,534 samples submitted to the PVML, 1,086 (70.8%) were collected from dogs and 29.2% from feline patients. Otological swabs (n = 533, 49.1%) were the most frequent samples from dogs, while cat urine samples (n = 384, 84.8%) predominated by far. Considering the canine samples, no bacterial growth (NBG) was observed in 443 (40.8%) samples, while only one colony type was noted in 516 (47.5%) samples. Gram-positive bacteria (n = 298) were more frequent than gram-negative bacteria (n = 77) in the skin. In urine samples, gram-negative bacteria (n = 94) were isolated more frequently than gram-positive bacteria (n = 47). In feline samples, a high number of NBG (n = 308, 68%) was observed. Gram-positive (n = 22) was predominant in comparison to gram-negative bacteria (n = 9) in cultures from the ear and skin swabs. Enterococcus spp. and Escherichia coli were the most frequently identified bacteria in urine samples. Among the Staphylococcus sp. strains of any origin, AMR frequency varied from 4.22% (amikacin) to 50.70% (sulfa/trimethoprim). Enterococcus spp. showed AMR frequencies from 12.5% (amoxicillin/clavulanic acid) to 62.06% (enrofloxacin). Among the gram-negative genera, E. coli presented AMR frequencies from 10.20% (gentamicin) to 60.0% (neomycin). The frequency of AMR was stable over time, and a profile of much higher resistance to fluoroquinolones in comparison to beta-lactams was observed. Discussion: The recurrence of skin and urinary infections implies the need for frequent treatment with antibiotics, which exerts selection pressure for resistance and multidrug resistance. In this study, the frequency of multidrug resistance was low, and the resistance to the tested antimicrobials showed high variation. However, a trend of high resistance to the fluoroquinolone group was observed in contrast to the low resistance to beta-lactams. This trend was consistent among the isolated bacteria, regardless of the type of sample or origin. The overprescription of fluoroquinolones in small animal practices has been widely documented in several countries. However, this class of antimicrobials, is highly prioritized for the treatment of infections in humans. Therefore, the selection of resistant strains has gained special emphasis, especially when considering the possibility of the transmission of resistant bacteria between pets and humans. In summary, the results of bacteriological tests conducted at the PVML-Universidade Federal do Rio Grande do Sul confirmed that ubiquitous bacteria predominate in clinical samples of dogs and cats. The high frequency of resistance to the fluoroquinolone group, while a predominance of susceptible strains in the first-choice drugs such as amoxicillin/clavulanic acid, may indicate excessive and empirical use of the second-choice drugs in clinical practice.

Isolation of Escherichia coli and Staphylococcus aureusin raw milk from refrigeration tanks: identification and antimicrobial resistance profiles / Isolamento de Escherichia coli e Staphylococcus aureus em leite cru de tanques de refrigeração: identificação e perfis de resistência antimicrobiana

Escherichia coli and Staphylococcus aureus in milk cooling tank reflects a hygienic deficit in animal management, production environment, and milk obtainment. With implications for public health as agents of infection and food poisoning, and the presence of antimicrobial-resistant strains. Therefore, were investigated in cooling tanks with high counts of somatic cells and total bacteria in milk. Microorganisms, in which a profile of resistance to antimicrobials was investigated, and whether there was a similarity in this profile between the strains of the eight dairy properties.Therefore, eighty-eight samples were obtained, and inoculated on Compact Dry® plates. Of this total, 27.27% (24/88) samples tested positive for E. coli and 56.81% (50/88) for S. aureus. Among 24 E. coli strains subjected to disk-diffusion antibiograms, 70.83% were resistant to rifampicin, 50% to ampicillin and 41.67% to cefoxitin and erythromycin, while of the 51 S. aureus strains, 94.32% expressed resistance to azetroanam, 86.27% to ampicillin and nalidixic acid, 76.47% to rifampicin and 47.06 % to erythromycin and cefoxitin. A criterion of resistance to over three antibiotics was observed for 8.33% (2/24) of the isolated E. coli strains and 17.65% (9/51) of the S. aureus strains, characterizing them as multidrug resistant (MDR) strains. Resistance phenotypes displayed high similarity between properties F5 and F6 for S. aureus, and properties F6 and F8 for E. coli when applying the Jaccard index. The presence of these antibiotic-resistant pathogenic microorganisms indicate flaws in milk production handling and sanitary conditions, representing risk to milk consumers.(AU)

Escherichia coli e Staphylococcus aureus no tanque de resfriamento de leite refletem um déficit higiênico no manejo animal, ambiente de produção e obtenção de leite. Com implicações para a saúde pública como agentes de infecção e intoxica-ção alimentar e presença de cepas resistentes a antimicrobianos. Portanto, foram investigados em tanques de resfriamento com altas contagens de células somáticas e bactérias totais no leite. Microrganismos, nos quais foi investigado o perfil de resistência aos antimicrobianos e se havia semelhança nesse perfil entre as cepas das oito propriedades leiteira. Para tanto, oitenta e oito amostras foram coletadas inoculadas em placas de Compact Dry®, destas 27,27% (24/88) amostras revelaram contaminação com E. coli, e 56,81% (50/88) com S. aureus. Entre 24 cepas de E. coli submetidas a antibiograma por disco-difusão 70,83% foram resistentes a rifampicina, 50% ampicilina, 41,67% cefoxitina e eritromicina, enquanto das 51 cepas de S. aureus 94,32% expressaram resistência a azetroanam, 86,27% a ampicilina e nalidíxico ácido, 76,47 % a rifampicina, e 47,06% a cefoxitina e eritromicina. O critério de resistência a mais de três antibióticos caracterizou 8,33% (2/24) das cepas de E. coli, e 17,65% (9/51) de S. aureus como multidroga resistente (MDR). O fenótipo resistência mostrou elevada similaridade entre as proprie-dades (F5 e F6) para S. aureus, e das propriedades (F6 e F8) para E. coli aplicando-se o índice de Jaccard. Estes microrganis-mos patogênicos resistentes a antibióticos, indicam falhas no manuseio e obtenção higiênica do leite, e que estes representam risco aos consumidores do leite.(AU)

Short chain organic acids in poultry: Literature review

The importance of Brazil for the International Trade of poultry products induces legislators and interested companies to pay special attention to complying with international standard quality requirements. One of these requirements is the suppression of the indiscriminate use of antibiotics in poultry, as in the strategy of their use as a growth promoter. To meet this demand, the study of short-chain organic acids becomes interesting and their review essential. As good examples for the issue described, short chain organic acids such as citric acid, fumaric acid, sorbic acid and malic acid are extremely important due to their antimicrobial qualities, accessibility, and international acceptance. In addition to the individual effect, the blend of acids, that is, the association of acids for the role of growth promoter or performance enhancer, combines acids that act synergistically, enhancing their qualities. This study aims to address the physicochemical characteristics, effects, and qualities of short-chain organic acids, as well as their associated effects, encouraging further work that fills the scientific gaps in empirical knowledge about these additives.(AU)

Antimicrobial resistance and mecA gene in Staphylococcus spp. isolates from horses used in family farming / Resistência antimicrobiana e gene mecA em Staphylococcus spp. isolados de cavalos usados na agricultura familiar

Horses can contribute to the spread of bacterial diseases, which can be caused mainly by, Staphylococcus spp., which are part of the animals' commensal microbiota, but it is also considered a pathogenic microorganism capable of causing serious infections. vancomycin, when it is resistant to methicillin. Antimicrobial resistance is considered a major health problem by the World Health Organization and the emergence of the mecA gene, responsible for resistance to the class of beta-lactam antibiotics. Thus, the aim of this work was to investigate the antimicrobial resistance profile and the presence of the mecA gene in Staphylococcus spp. isolated from the nasal, oral and auricular microbiota of horses used as animal traction on small family farms. Nasal, oral and auricular swabs were collected from 38 horses, with 29 (76.3%) isolated in nasal swab, 15 (39.4%) in auricular swab and 9 (23.6%) in oral swab, totaling 53 Staphylococcus spp. and 50 (94.33%) samples were resistant to the 11 antimicrobials tested, none of which were positive for molecular tests to identify the mecA gene. The results demonstrate the presence of Staphylococcus spp. associated with high (94.33%) bacterial resistance, indicating that horses can be considered reservoirs of multi-resistant microorganisms.

Os equinos podem contribuir para a disseminação de doenças bacterianas, podendo ser causadas principalmente pelo, Staphylococcus spp., que fazem parte da microbiota comensal dos animais, mas também é considerado microrganismo patogênico capaz de causar infecções graves, em seu tratamento o medicamento mais utilizado é a vancomicina, quando há resistência a meticilina. A resistência antimicrobiana é considerada um dos principais problemas de saúde pela Organização Mundial de Saúde e o surgimento do gene mecA, responsável pela resistência à classe dos antibióticos beta-lactâmicos. Deste modo, o objetivo deste trabalho foi investigar o perfil de resistência antimicrobiana e a presença do gene mecA em Staphylococcus spp. isoladas da microbiota nasal, oral e auricular de cavalos usados como tração animal em pequenas propriedades familiares. Foram coletados swabs nasal, oral e auricular de 38 cavalos, sendo identificados 29 (76,3%) isolados em swab nasal, 15 (39,4%) em swab auricular e 9 (23,6%) em swab oral, totalizando 53 Staphylococcus spp. e 50 (94,33%) amostras foram resistentes aos 11 antimicrobianos testados, nenhuma amostra foi positiva aos testes moleculares para identificação do gene mecA. Os resultados demonstram a presença de Staphylococcus spp. associada à alta (94,33%) resistência bacteriana, indicando que os cavalos podem ser considerados reservatórios de microrganismos multirresistentes.