RESUMO
Samples from a dairy cattle waste-fed anaerobic digester were collected across seasons to assess sanitary safety for biofertilizer use. Isolated enterobacteria (suggestive of Escherichia coli) were tested for susceptibility to biocides, antimicrobials, and biofilm-forming capability. Results revealed a decrease in total bacteria, coliforms, and enterobacteria in biofertilizer compared to the effluent. Among 488 isolates, 98.12 % exhibited high biofilm formation. Biofertilizer isolates exhibited a similar biofilm formation capability as effluent isolates in summer, but greater propensity in winter. Resistance to biocides and antimicrobials varied, with tetracycline resistance reaching 19 %. Of the isolates, 25 were multidrug-resistant (MDR), with 64 % resistant to three drugs. Positive correlations were observed between MDR and increased biofilm formation capacity in both samples, while there was negative correlation between MDR and increased biocide resistance. A higher number of MDR bacteria were found in biofertilizer compared to the effluent, revealing the persistence of E. coli resistance, posing challenges to food safety and public health.
Assuntos
Anti-Infecciosos , Desinfetantes , Saúde Única , Animais , Bovinos , Escherichia coli , Enterobacteriaceae , Virulência , Anaerobiose , Águas Residuárias , Antibacterianos , Desinfetantes/farmacologia , Testes de Sensibilidade MicrobianaRESUMO
Cheeses are dairy products that can potentially contain a diverse range of harmful bacteria that could be consumed by humans, including the enteric pathogen Escherichia coli. This study aimed to characterize the presence of total coliforms, assess the antimicrobial susceptibility profiles of the main commercial antimicrobial classes and biocides, and evaluate the ability of 50 E. coli isolates obtained from fresh cheese sold in the southern region of Espírito Santo, Brazil, to produce biofilms. The counts of total coliforms + E. coli obtained averages of (A) 7.22 × 106 CFU/g, (B) 9.35 × 107 CFU/g, and (C) 1.16 × 106 CFU/g for different brands. All isolates were capable of forming biofilms, with 8%, 76%, and 16% of these isolates presenting high, moderate, and low adherence in biofilm formation, respectively. Most strains showed inhibition halos for the biocides chlorhexidine digluconate 2% (16 mm ± 4.34), iodopovidone 10% (7.14 mm ± 0.36), and sodium hypochlorite 2% (7.12 mm ± 0.33). Out of the 50 strains, 21 (42%) were resistant to at least one of the antimicrobials. Regarding the multiple resistance index, 3 (6%) strains were resistant to 3 or more antimicrobial classes. Furthermore, 2 (4%) were extended-spectrum beta-lactamases producers. Resistance to ampicillin and amoxicillin was observed in 20% and 40% of the strains, respectively. In contrast, gentamicin was the most effective antimicrobial, with a sensitivity rate of 100%. The findings indicate that E. coli present in fresh cheese may possess unique physiological characteristics that could be associated with their persistence, virulence, and multidrug resistance. These results raise significant public health concerns since contaminated food can pose risks to consumers' health, emphasizing the importance of reinforcing hygienic-sanitary controls at all stages of production.
Assuntos
Queijo , Desinfetantes , Humanos , Animais , Escherichia coli , Antibacterianos/farmacologia , Queijo/microbiologia , Fatores de Virulência , Farmacorresistência Bacteriana , Leite/microbiologia , Desinfetantes/farmacologiaRESUMO
Polymeric films containing pomegranate peel extract (PPE) can act as a drug-delivery platform for topical treatment of candidiasis. The composition, mechanical resistance, and in vitro antifungal activity of a polymeric film containing PPE at 1.25 mg.mL-1 were investigated. Films were prepared using a solvent casting technique. The incorporation of PPE in the polymeric matrix gave rise to homogeneous, smooth, shiny, and yellowish-brown films. FTIR spectra of the film containing PPE showed differences without compromising the stability of the extract and the matrix. SEM analysis showed the existence of interruptions in the continuity of the films with extract, which promoted a reduction in the mechanical parameters without significantly changing the tensile strength and elongation at break. Films showed adequate mechanical properties and antifungal activity against Candida albicans, C. glabrata, C. krusei and C. tropicalis.
Assuntos
Candidíase , Punica granatum , Antifúngicos/farmacologia , Candida albicans , Polímeros , Extratos Vegetais/farmacologia , Candidíase/tratamento farmacológicoRESUMO
Introduction: Staphylococcus aureus and Escherichia coli are pathogens that cause chronic infections due to antibiotic resistance mechanisms and their ability to adhere to surfaces and to form biofilms. The search for new agents from natural resources to counter microbial biofilms is an urgent priority in healthcare. Objetive: chemical composition, antibacterial and anti-biofilm activity of Plectranthus amboinicus essential oil (PAEO) and carvacrol were investigated against E. coli and S. aureus. Methodology: PAEO was chemically analyzed using gas chromatography coupled to a mass spectrometer. Antimicrobial activity was assessed by the disc diffusion method and broth microdilution method to determine minimum inhibitory concentrations (MICs). Antibiofilm activity was investigated using 96-well plates with a crystal violet assay. Results: carvacrol (85.25%) was the major component of PAEO. The disc diffusion test confirmed the ability of PAEO and carvacrol in inhibiting bacteria in their planktonic form. The MICs of PAEO against S. aureus and E. coli were 0.31 and 1.25%, respectively, with bactericidal effect. Carvacrol demonstrated a significant antibacterial property (MIC = 0.31%), exhibiting bacteriostatic effects against S. aureus and bactericidal effects against E. coli. Carvacrol considerably inhibited E. coli biofilm formations (58.9%). Moreover, carvacrol inactivated the mature biofilms. Conclusion: the data obtained are promising, and facilitates the development of new therapeutic alternatives. These results indicate the potential of carvacrol in treating diseases caused by E. coli and S. aureus
Introdução: Escherichia coli e Staphylococcus aureus são patógenos que podem causar infecções persistentes devido aos mecanismos de resistência e à sua capacidade de aderir e formar biofilme. A busca por novos agentes a partir de recursos naturais para combater o biofilme é uma prioridade na área da saúde. Objetivo: a composição química, a atividade antibacteriana e anti-biofilme do óleo essencial de Plectranthus amboinicus (PAEO) e do carvacrol foram investigadas frente a S. aureus e E. coli. Metodologia: a análise química do PAEO foi realizada por cromatografia gasosa acoplada a espectrômetro de massa. A atividade antimicrobiana foi avaliada por difusão em disco e microdiluição em caldo para determinar as concentrações inibitórias mínimas (CIMs). As atividades do anti-biofilme foram investigadas usando placas de 96 poços pelo ensaio de cristal violeta. Resultados: o carvacrol (85,25%) foi o principal componente do PAEO. A capacidade do PAEO e do carvacrol para inibir bactérias planctônica foi confirmada por difusão em disco. A CIM do PAEO para S. aureus e E. coli foi de 0,31 e 1,25%, respectivamente, com efeito bactericida. O carvacrol demonstrou uma propriedade antibacteriana significativa (CIM=0,31%), exibindo efeito bacteriostático frente a S. aureus e efeito bactericida para E. coli. O carvacrol inibiu significativamente a formação de biofilme de E. coli (58,9%) e promoveu a desestabilização do biofilme maduro. Conclusão: os dados obtidos são promissores, possibilitando o desenvolvimento de novas alternativas terapêuticas e revelam a potencialidade do carvacrol como fonte para o tratamento de doenças causadas por E. coli e S. aureus.
Assuntos
Staphylococcus aureus/efeitos dos fármacos , Óleos Voláteis/farmacologia , Biofilmes/efeitos dos fármacos , Plectranthus/química , Escherichia coli/efeitos dos fármacos , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana/métodosRESUMO
OBJECTIVE: An experimental infection based on a tissue cage model was reproduced to evaluate the interference subinhibitory concentration (SIC) of metronidazole in Bacteroides fragilis OMV production patterns and immunological and histological characteristics of the host facing the experimental challenge. METHODS: A tissue cage model was reproduced for B. fragilis experimental challenge in three Wistar rats groups: negative control group (NC) without bacterial inoculation; positive control group (PC) infected with parental strain; and experimental group (EG) infected with the parental strain and treated with metronidazole SIC. Tissue cage sections and histological preparations were evaluated under optical and transmission electron microscope. Observations included OMV identification and count and cellular envelope evaluation. Transcriptional analyses were performed to evaluate cytokines expression levels. RESULTS: Total counts of leukocytes and neutrophils were higher for EG, and slight increase in PC group. It was observed an exacerbated inflammatory infiltrate after 8 days on infection. The expression of TNF-α was increased during the experiments, along with IL-1α and IL-6. MCP-1 levels were suppressed in almost every evaluated time-point. The IL-10 was exacerbated in EG group. A massive production and release of OMV and cell wall thickening were observed especially the EG group. CONCLUSIONS: Despite literature data suggest positive association between OMV and antimicrobial stress for Gram negatives, no correlations are made for B. fragilis and drug-response during experimental model of infection. Results corroborate observations in which OMV may be involved in bacterial pathogenicity once the phenomenon was observed along histological evidence of exacerbated inflammation and cytokines modulation.
Assuntos
Infecções Bacterianas , Bacteroides fragilis , Animais , Antibacterianos/farmacologia , Infecções Bacterianas/tratamento farmacológico , Metronidazol/farmacologia , Ratos , Ratos WistarRESUMO
Chronic wounds constitute a serious public health problem, and developing pharmaceutical dosage forms to ensure patient comfort and safety, as well as optimizing treatment effectiveness, are of great interest in the pharmaceutical, medical and biomaterial fields. In this work, the preparation of films based on blends of poly(vinyl alcohol), starch and poly(acrylic acid), polymers widely used as pharmaceutical excipients, and pomegranate peel extract (PPE), a bioactive compound with antimicrobial and healing activities relevant to the use as a bioactive wound dressing, was proposed. Initially, the minimum inhibitory concentration (MIC) of the PPE was investigated by an in vitro method. Then, the best concentration of the PPE to be used to prepare the films was researched using an antimicrobial susceptibility test with the disc diffusion method. The microbiological assay was performed in films prepared by the solvent casting method in the presence of two concentrations of PPE: 1.25% w/v and 2.5% w/v. Films containing the lower PPE concentration showed antimicrobial activity against Staphylococcus aureus and Staphylococcus epidermidis, with a difference that was not considered statistically significant when compared to the higher concentration of the extract. Therefore, the films prepared with the lower proportion of PPE (1.25% w/v) were considered for the other studies. The miscibility and stability of the extract in the films were investigated by thermal analysis. Parameters that determine the barrier properties of the films were also investigated by complementary techniques. Finally, in vitro biological tests were performed for safety evaluation and activity research. Analysis of the results showed that the incorporation of the higher proportion of starch in the blend (15% v/v) (PVA:S:PAA:PPE4) yielded smooth, transparent, and domain-free films without phase separation. Additionally, the PVA:S:PAA:PPE4 film presented barrier properties suitable for use as a cover. These films, when subjected to the in vitro hemolytic activity assay, were nonhemolytic and biocompatible. No toxicity from the extract was observed at the concentrations studied. The results of the wound healing in vitro test showed that films containing 1.25% PPE are efficient in reducing the scratch open area, provoking almost total closure of the scratches within 48 h without cytotoxicity.
Assuntos
Antibacterianos/química , Bandagens , Membranas Artificiais , Álcool de Polivinil/química , Punica granatum/química , Amido/química , Animais , Linhagem Celular , Camundongos , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus epidermidis/crescimento & desenvolvimentoRESUMO
Although lifestyle and physiology in obese individuals are accepted to lead to changes in the intestinal microbiota, uncertainty remains about microbiota dysbiosis, and xenobiotics intake, as a source of selective pressure, independent of antimicrobial chemotherapy. The aim of this study was to compare the occurrence of antimicrobial resistance genetic markers (ARG) in faecal specimens of eutrophic, overweight and obese individuals, and their correlation with xenobiotic intake and gut bacteria density. Methods: This was a cross-sectional case-controlled study including 72 adult participants with no record of intestinal or systemic diseases, or recent use of antimicrobials, grouped as eutrophic, overweight, or obese. Anthropometric profile, eating habits and oral xenobiotics intake were recorded. Faecal metagenomic DNA was used to screen for ARG by PCR, and to measure bacterial groups by fluorescence in situ hybridization (FISH). Student's t and Wilcoxon tests were used to compare means and differences in ARG detection (95% confidence intervals). Correlation analyses (odds ratio) and relationships between bacteria density and ARG were determined. Results: Increase in abdominal circumference, waist circumference, hip, waist-hip ratio, BMI, carbohydrate, fibres, and total calorie intakes were different from eutrophic to obese participants. Habitual use of antihypertensive and anti-inflammatory drugs, antacids, and artificial sweeteners were associated mainly with obesity and overweight. Nutritional supplements were associated to the eutrophic group. ARG screening showed differences being more frequent among obese, and positive for 27 genetic markers related to ß-lactams, tetracyclines, the macrolide lincosamide and streptogramin group, quinolones, sulfonamides, aminoglycosides, and efflux pump. Positive correlation between ARG and BMI, caloric intake, and intake of xenobiotics, was observed for obese individuals. Relationships among ARG detection and bacteria densities were also different. Conclusions: This study reinforces the hypothesis that obese individuals may harbour an altered gut microbiota, if compared to eutrophic. The overweight individuals display a transitional gut microbiota which seems to be between eutrophic and obese. Furthermore, the increased xenobiotic intake associated to obesity may play an important role in the antimicrobial resistance phenomenon.
Assuntos
Resistência Microbiana a Medicamentos/genética , Trato Gastrointestinal/microbiologia , Sobrepeso/microbiologia , Adulto , Antiácidos/uso terapêutico , Anti-Inflamatórios não Esteroides/uso terapêutico , Anti-Hipertensivos/uso terapêutico , Suplementos Nutricionais , Fezes/microbiologia , Feminino , Microbioma Gastrointestinal/genética , Humanos , Hibridização in Situ Fluorescente , Masculino , Pessoa de Meia-Idade , Edulcorantes/administração & dosagem , Xenobióticos/administração & dosagem , Adulto JovemRESUMO
Cooperation between researchers in the areas of medical, pharmaceutical and materials science has facilitated the development of pharmaceutical dosage forms that elicit therapeutic effects and protective action with a single product. In addition to optimizing pharmacologic action, such dosage forms provide greater patient comfort and increase success and treatment compliance. In the present work, we prepared semipermeable bioactive electrospun fibers for use as wound dressings containing silver sulfadiazine complexed with ß-cyclodextrin in a poly(Æ-caprolactone) nanofiber matrix aiming to reduce the direct contact between silver and skin and to modulate the drug release. Wound dressings were prepared by electrospinning, and were subjected to ATR-FT-IR and TG/DTG assays to evaluate drug stability. The hydrophilicity of the fibrous nanostructure in water and PBS buffer was studied by goniometry. Electrospun fibers permeability and swelling capacity were assessed, and a dissolution test was performed. In vitro biological tests were realized to investigate the biological compatibility and antimicrobial activity. We obtained flexible matrices that were each approximately 1.0 g in weight. The electrospun fibers were shown to be semipermeable, with water vapor transmission and swelling indexes compatible with the proposed objective. The hydrophilicity was moderate. Matrices containing pure drug modulated drug release adequately during 24 h but presented a high hemolytic index. Complexation promoted a decrease in the hemolytic index and in the drug release but did not negatively impact antimicrobial activity. The drug was released predominantly by diffusion. These results indicate that electrospun PCL matrices containing ß-cyclodextrin/silver sulfadiazine inclusion complexes are a promising pharmaceutical dosage form for wound healing.
Assuntos
Portadores de Fármacos/síntese química , Nanofibras/química , Poliésteres/química , Sulfadiazina de Prata/administração & dosagem , Cicatrização , beta-Ciclodextrinas/administração & dosagem , Bandagens , Células Sanguíneas/efeitos dos fármacos , Células Sanguíneas/fisiologia , Fenômenos Químicos , Formas de Dosagem , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Galvanoplastia , Hemólise/efeitos dos fármacos , Humanos , Teste de Materiais , Testes de Sensibilidade Microbiana , Sulfadiazina de Prata/química , Termogravimetria , Cicatrização/efeitos dos fármacos , beta-Ciclodextrinas/químicaRESUMO
Although most Brazilian dairy products meet high technological standards, there are quality issues regarding milk production, which may reduce the final product quality. Several microbial species may contaminate milk during manufacture and handling. If antimicrobial usage remains uncontrolled in dairy cattle, the horizontal transfer of antimicrobial resistance genes in foodstuffs may be of particular concern for both food producers and dairy industry. This study focused on the evaluation of putative Gram positive cocci in Minas cheese and of antimicrobial and biocide resistance genes among the isolated bacteria. Representative samples of 7 different industrially trademarked Minas cheeses (n = 35) were processed for selective culture and isolation of Gram positive cocci. All isolated bacteria were identified by DNA sequencing of the 16S rRNA gene. Antimicrobial resistance genes were screened by PCR. Overall, 208 strains were isolated and identified as follows: Enterococcus faecalis (47.6%), Macrococcus caseolyticus (18.3%), Enterococcus faecium (11.5%), Enterococcus caseliflavus (7.7%), Staphylococcus haemolyticus (7.2%), Staphylococcus aureus (4.3%), Staphylococcus epidermidis (2.9%), and Enterococcus hirae (0.5%). The genetic markers mecA (78.0%) and smr (71.4%) were the most prevalent, but others were also detected, such as blaZ (65.2%), msrA (60.9%), msrB (46.6%), linA (54.7%), and aacA-aphD (47.6%). The occurrence of opportunist pathogenic bacteria harboring antimicrobial resistance markers in the cheese samples are of special concern, since these bacteria are not considered harmful contaminating agents according to the Brazilian sanitary regulations. However, they are potentially pathogenic bacteria and the cheese may be considered a reservoir for antimicrobial resistance genes available for horizontal transfer through the food chain, manufacturing personnel and consumers.
Assuntos
Queijo/microbiologia , Farmacorresistência Bacteriana/genética , Cocos Gram-Positivos/genética , Cocos Gram-Positivos/isolamento & purificação , Animais , Antibacterianos , Brasil , Bovinos , DNA Bacteriano/análise , Enterococcus/genética , Enterococcus/isolamento & purificação , Microbiologia de Alimentos , Marcadores Genéticos , Cocos Gram-Positivos/efeitos dos fármacos , Leite/microbiologia , RNA Ribossômico 16S/genética , Staphylococcus/genética , Staphylococcus/isolamento & purificaçãoRESUMO
Nonfermenting Gram-negative bacteria such as Pseudomonas aeruginosa and Acinetobacter baumannii are widespread in the environment and are increasingly associated with nosocomial infections, often associated with multidrug-resistance phenotypes. This study aimed to evaluate epidemiological, physiological, and molecular characteristics of carbapenem resistance in P. aeruginosa and A. baumannii. In total, 63 nonreplicated strains (44 A. baumannii and 19 P. aeruginosa) were isolated from hospitalized patients. Antimicrobial resistance patterns, biocide tolerance, oxidative stress, hemolytic activity, and biofilm formation were assessed. Genetic markers related to ß-lactamase synthesis, efflux systems, and porin loss were screened by PCR. Epidemiological data of patients were analyzed. Advanced age, intensive care unit admission, invasive medical devices, treatment with fluoroquinolones or ß-lactams/ß-lactamase inhibitor combinations, and prolonged hospital stay were predisposing factors for infection. Colistin showed to be active in vitro against these bacteria. Carbapenem-resistant P. aeruginosa strains did not show hemolytic activity and were less tolerant to oxidative stress and biocides. However, increased ability of biofilm formation was observed, comparing to the carbapenem-susceptible isolates. Genetic markers related to oxacillinases synthesis (OXA-23 and OXA-143), oprD absence, and efflux pump (adeB) were detected in carbapenem-resistant A. baumannii. Screening for OXA-51-like gene was performed as confirmatory test for A. baumannii identification. In P. aeruginosa genes encoding efflux pumps (MexAB-OprM, MexCD-OprJ, MexEF-OprN, and MexXY-OprM) and SPM-1 were found; besides, oprD absence was also observed. Our results suggest that these organisms are well adapted to different environments and confirm the difficulty of therapeutic management of patients with infections associated with multidrug-resistant microorganisms, with direct impact on mortality and epidemiological control of these strains in health centers.
Assuntos
Acinetobacter baumannii/genética , Farmacorresistência Bacteriana Múltipla/genética , Genes MDR , Porinas/genética , Pseudomonas aeruginosa/genética , beta-Lactamases/genética , Infecções por Acinetobacter/tratamento farmacológico , Infecções por Acinetobacter/epidemiologia , Infecções por Acinetobacter/microbiologia , Acinetobacter baumannii/classificação , Acinetobacter baumannii/efeitos dos fármacos , Acinetobacter baumannii/isolamento & purificação , Adolescente , Adulto , Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Brasil/epidemiologia , Carbapenêmicos/farmacologia , Criança , Pré-Escolar , Estudos Transversais , Feminino , Fluoroquinolonas/farmacologia , Expressão Gênica , Hospitais , Humanos , Lactente , Recém-Nascido , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Epidemiologia Molecular , Estresse Oxidativo , Porinas/metabolismo , Infecções por Pseudomonas/tratamento farmacológico , Infecções por Pseudomonas/epidemiologia , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/classificação , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/isolamento & purificação , beta-Lactamases/metabolismoRESUMO
INTRODUCTION: Bacterial resistance is a growing concern in the nosocomial environment in which Klebsiella pneumoniae and Enterobacter aerogenes play an important role due to their opportunism and carbapenemase-production. This work aimed to evaluate physiological and molecular characteristics of carbapenem-resistant K. pneumoniae and E. aerogenes isolated in a Brazilian tertiary hospital. METHODOLOGY: In total, 42 carbapenem-resistant bacteria isolated from clinical specimens were included (21 K. pneumoniae and 21 E. aerogenes). Drug-sensitive K. pneumoniae (n = 27) were also included. Antimicrobial susceptibility and biocide tolerance patterns, hemolytic activity, tolerance to oxidative stress, and aggregative ability were assessed. Genetic markers related to carbapenem resistance, or ESBL-production were screened by PCR. RESULTS: Compared to drug-sensitive strains, carbapenem-resistant K. pneumoniae were more tolerant to biocides and to oxidative stress, and they displayed an increase in biofilm formation. The genetic markers blaKPC (95.2%) and blaTEM (90.5%) were the most frequent. Among the carbapenem-resistant E. aerogenes strains, blaKPC, and blaTEM were detected in all bacteria. Drug-sensitive E. aerogenes were not isolated in the same period. blaSHV, blaVIM, and blaCTX markers were also observed among carbapenem-resistant bacteria. CONCLUSIONS: Results suggest that carbapenemase-producing enterobacteria might show peculiar characteristics regarding their physiology associated with their environmental persistency, virulence, and multidrug resistance. The observed phenomenon may have implications not only for antimicrobial chemotherapy, but also for the prognosis of infectious diseases and infection control.
Assuntos
Antibacterianos/farmacologia , Carbapenêmicos/farmacologia , Enterobacter aerogenes/efeitos dos fármacos , Enterobacter aerogenes/genética , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/genética , Resistência beta-Lactâmica , Adulto , Idoso , Idoso de 80 Anos ou mais , Biofilmes/crescimento & desenvolvimento , Brasil , Estudos Transversais , Desinfetantes/farmacologia , Enterobacter aerogenes/fisiologia , Infecções por Enterobacteriaceae/microbiologia , Feminino , Humanos , Klebsiella pneumoniae/fisiologia , Masculino , Pessoa de Meia-Idade , Estresse Oxidativo , Reação em Cadeia da Polimerase , Estresse Fisiológico , Centros de Atenção Terciária , Adulto JovemRESUMO
Anaerobic digestion is an alternative method for the treatment of animal manure and wastewater. The anaerobic bioconversion of biomass requires a multi-step biological process, including microorganisms with distinct roles. The diversity and composition of microbial structure in pilot-scale anaerobic digestion operating at ambient temperature in Brazil were studied. Influence of the seasonal and temporal patterns on bacterial and archaeal communities were assessed by studying the variations in density, dynamic and diversity and structure. The average daily biogas produced in the summer and winter months was 18.7 and 16 L day(-1), respectively, and there was no difference in the average methane yield. Quantitative PCR analysis revealed that no differences in abundances and dynamics were found for bacterial communities and the total number of Archaea in different seasons. Analysis of bacterial clone libraries revealed a predominance of Firmicutes (54.5 %/summer and 46.7 %/winter) and Bacteroidetes (31.4 %/summer and 44.4 %/winter). Within the Archaea, the phylum Euryarchaeota was predominant in both digesters. Phylogenetic distribution showed changes in percentage between the phyla identified, but no alterations were recorded in the quality and amount of produced methane or community dynamics. The results may suggest that redundancy of microbial groups may have occurred, pointing to a more complex microbial community in the ecosystem related to this ambient temperature system.
Assuntos
Bactérias/metabolismo , Biodiversidade , Esterco/microbiologia , Metano/metabolismo , Anaerobiose , Animais , Bactérias/classificação , Bactérias/isolamento & purificação , Brasil , Bovinos , Esterco/análise , Metano/análise , FilogeniaRESUMO
Anaerobic digestion figures as a sustainable alternative to avoid discharge of cattle manure in the environment, which results in biogas and biofertilizer. Persistence of potentially pathogenic and drug-resistant bacteria during anaerobic digestion of cattle manure was evaluated. Selective cultures were performed for enterobacteria (ENT), non-fermenting Gram-negative rods (NFR) and Gram-positive cocci (GPC). Antimicrobial susceptibility patterns were determined and a decay of all bacterial groups was observed after 60days. Multidrug-resistant bacteria were detected both the influent and effluent. GPC, the most prevalent group was highly resistant against penicillin and levofloxacin, whereas resistance to ampicillin, ampicillin-sulbactam and chloramphenicol was frequently observed in the ENT and NFR groups. The data point out the need of discussions to better address management of biodigesters and the implementation of sanitary and microbiological safe treatments of animal manures to avoid consequences to human, animal and environmental health.
Assuntos
Bactérias/crescimento & desenvolvimento , Resistência Microbiana a Medicamentos , Esterco/microbiologia , Eliminação de Resíduos/métodos , Anaerobiose/efeitos dos fármacos , Animais , Antibacterianos/farmacologia , Bactérias/isolamento & purificação , Biodegradação Ambiental/efeitos dos fármacos , Biocombustíveis/microbiologia , Reatores Biológicos/microbiologia , Bovinos , Contagem de Colônia Microbiana , Humanos , Testes de Sensibilidade Microbiana , Projetos Piloto , Especificidade da EspécieRESUMO
Coagulase-negative staphylococci (CoNS), which are generally neglected as foodborne bacteria, are emerging as significant opportunistic pathogens that may be highly resistant to available antimicrobial drugs. In this study, antimicrobial susceptibility patterns, mecA gene occurrence, and virulence-associated characteristics were evaluated in CoNS isolated from soft cheese in Brazil. A total of 227 bacterial isolates were recovered from 35 cheese samples belonging to 5 batches with 7 different trademarks. The CoNS counts ranged from 10(6) to 10(7) CFU/g. High antimicrobial resistance percentages were observed for oxacillin (76.2%), penicillin (78.5%), erythromycin (67.8%), gentamicin (47.2%), clindamycin (35.7%), rifampicin (26.8%), azithromycin (14.7%), tetracycline (14.7%), levofloxacin (14.2%), and sulfamethoxazole-trimethoprim (11.9%). A low antimicrobial resistance percentage was observed for chloramphenicol (2.3%), and all of the tested bacteria were susceptible to vancomycin and linezolid. In total, a multiple antibiotic resistance (MAR) index of >0.2 was observed for 80.6% of the isolated CoNS. However, the MAR index ranged from 50% to 92.6% when only bacterial cheese isolates belonging to the same trademark were considered. Regarding to the prevalence of CoNS carrying mecA gene, 81.5% of the isolated strains were mecA(+) , and 76.2% of these were phenotypically resistant to oxacillin. Three isolates carried the enterotoxin A gene (sea), 29.5% produced biofilm in a laboratory test, and α- or ß-hemolysis were observed for 3% and 5.2%, respectively. This study highlights the extent of the antimicrobial resistance phenomenon in neglected foodborne microorganisms and the potential public health risks that are related to the consumption of CoNS-contaminated soft cheese.
Assuntos
Proteínas de Bactérias/genética , Queijo/microbiologia , Resistência Microbiana a Medicamentos , Staphylococcus/isolamento & purificação , Staphylococcus/patogenicidade , Antibacterianos/farmacologia , Proteínas de Bactérias/metabolismo , Brasil , Coagulase/metabolismo , Contagem de Colônia Microbiana , DNA Bacteriano/genética , Contaminação de Alimentos , Microbiologia de Alimentos , Análise de Sequência de DNA , Staphylococcus/efeitos dos fármacos , VirulênciaRESUMO
The use of antimicrobials and toxic metals should be considered carefully in aquaculture and surrounding environments. We aimed to evaluate medically relevant bacteria in an aquaculture system and their susceptibility to antimicrobials and toxic metals. Selective cultures for enterobacteria (ENT), non-fermenting Gram-negative rods (NFR) and Gram-positive cocci (GPC) were obtained from water samples collected in two different year seasons. The isolated bacteria were biochemically identified and antimicrobial and toxic metal susceptibility patterns were determined. Overall, 407 representative strains were recovered. In general, bacteria isolated from fish ponds showed higher multiple antibiotic resistance indices when compared to those isolated from a water-fed canal. Resistance to penicillin and azithromycin was observed more frequently in the GPC group, whereas resistance to ampicillin and ampicillin/sulbactam or gentamicin was observed more frequently in the ENT and NFR groups, respectively. All the isolated bacteria were tolerant to nickel, zinc, chromium and copper at high levels (≥1,024 µg mL(-1)), whereas tolerance to cadmium and mercury varied among the isolated bacteria (2-1,024 µg mL(-1)). Multidrug-resistant bacteria were more frequent and diverse in fish ponds than in the water-fed canal. A positive correlation was observed between antimicrobial resistance and metal tolerance. The data point out the need for water treatment associated with the aquaculture system.