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This study aimed to identify contamination sources in raw milk and cheese on small farms in Brazil by isolating Escherichia coli at various stages of milk production and cheese manufacturing. The study targeted EAEC, EIEC, ETEC, EPEC, STEC, and ExPEC pathotypes, characterizing isolates for the presence of virulence genes, phylogroups, antimicrobial susceptibility, and phylogenetic relationships using PFGE and MLST. The presence of antimicrobial resistance genes and serogroups was also determined. Three categories of E. coli were identified: pathogenic, commensal, and ceftriaxone-resistant (ESBL) strains. Pathogenic EPEC, STEC, and ExPEC isolates were detected in milk and cheese samples. Most isolates belonged to phylogroups A and B1 and were resistant to antimicrobials such as nalidixic acid, ampicillin, kanamycin, streptomycin, sulfisoxazole, and tetracycline. Genetic analysis revealed that E. coli with identical virulence genes were present at different stages within the same farm. The most frequently identified serogroup was O18, and MLST identified ST131 associated with pathogenic isolates. The study concluded that E. coli was present at multiple points in milk collection and cheese production, with significant phylogroups and high antimicrobial resistance. These findings highlight the public health risk posed by contamination in raw milk and fresh cheese, emphasizing the need to adopt hygienic practices to control these microorganisms.
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BACKGROUND: Pink coloration of breast milk is uncommon and it´s associated with colonization by Serratia marcescens, which is most frequently isolated in intensive care settings. Misinterpretation of the pink coloration may lead to premature cessation of breastfeeding. The objective is to present four cases of pink discoloration. METHODS: Two retrospective and two prospective cases of pink discoloration in breast milk are described, which were reported to the lead author. RESULTS: Four healthy mother-infant pairs with documented pink discoloration are presented. S. marcescens was isolated from breast milk samples. All four infants were asymptomatic and underwent enterobacteria cultures. The mothers received outpatient antibiotic treatment, and two infants received treatment as well. Subsequent cultures yielded negative results, and the pink discoloration ceased. All mothers successfully resumed breastfeeding. CONCLUSIONS: There are very few reported cases of pink breast milk in the global literature. Colonization by S. marcescens is not an indication for discontinuation of breastfeeding.
INTRODUCCIÓN: La coloración rosa de la leche materna es poco frecuente y está asociada a colonización por Serratia marcescens. Se aísla con mayor frecuencia en entornos de cuidados intensivos. La desinformación por la coloración rosa puede conducir a una terminación prematura de la lactancia. El objetivo es presentar cuatro casos de coloración rosa de la leche materna. MÉTODOS: Se describen dos casos retrospectivos y dos prospectivos de presentación de leche materna de color rosa. Los casos fueron reportados a la autora principal. RESULTADOS: Se presentan cuatro binomios sanos con reporte de coloración rosa. Se aisló S. marcescens en una muestra de leche materna. Los cuatro lactantes eran asintomáticos y tuvieron cultivos para la enterobacteria. Las madres fueron tratadas con antibiótico ambulatorio. Dos lactantes recibieron tratamiento. Todos los cultivos posteriores fueron negativos y la coloración rosa cesó. Todos reanudaron la lactancia materna de forma exitosa. CONCLUSIONES: Existen muy pocos casos de leche de color rosa reportados en la literatura mundial. La colonización por S. marcescens no es una indicación de suspensión de la lactancia.
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Antibacterianos , Lactancia Materna , Leche Humana , Infecciones por Serratia , Serratia marcescens , Adulto , Femenino , Humanos , Lactante , Recién Nacido , Antibacterianos/administración & dosificación , Leche Humana/microbiología , Estudios Prospectivos , Estudios Retrospectivos , Infecciones por Serratia/microbiología , Infecciones por Serratia/diagnóstico , Serratia marcescens/aislamiento & purificaciónRESUMEN
INTRODUCTION: The multifactorial etiology of gastroenteritis emphasizes the need for different laboratory methods to identify or exclude infectious agents and evaluate the severity of diarrheal disease. OBJECTIVE: To diagnose the infectious etiology in diarrheic children and to evaluate some fecal markers associated with intestinal integrity. MATERIALS AND METHODS: The study group comprised 45 children with diarrheal disease, tested for enteropathogens and malabsorption markers, and 76 children whose feces were used for fat evaluation by the traditional and acid steatocrit tests. RESULTS: We observed acute diarrhea in 80% of the children and persistent diarrhea in 20%. Of the diarrheic individuals analyzed, 40% were positive for enteropathogens, with rotavirus (13.3%) and Giardia duodenalis (11.1%) the most frequently diagnosed. Among the infected patients, occult blood was more evident in those carrying pathogenic bacteria (40%) and enteroviruses (40%), while steatorrhea was observed in infections by the protozoa G. duodenalis (35.7%). Children with diarrhea excreted significantly more lipids in feces than non-diarrheic children, as determined by the traditional (p<0.0003) and acid steatocrit (p<0.0001) methods. Moreover, the acid steatocrit method detected 16.7% more fecal fat than the traditional method. CONCLUSIONS: Childhood diarrhea can lead to increasingly severe nutrient deficiencies. Steatorrhea is the hallmark of malabsorption, and a stool test, such as the acid steatocrit, can be routinely used as a laboratory tool for the semi-quantitative evaluation of fat malabsorption in diarrheic children.
Introducción. La etiología multifactorial de la gastroenteritis enfatiza la necesidad de usar diferentes métodos de laboratorio para identificar o excluir agentes infecciosos y evaluar la gravedad de la enfermedad diarreica. Objetivo. Diagnosticar la etiología infecciosa de la diarrea en niños y evaluar algunos marcadores fecales asociados con la integridad intestinal. Materiales y métodos. Se estudiaron 45 niños con enfermedad diarreica, en los cuales se evaluaron la presencia de enteropatógenos y los marcadores de malabsorción. Se analizaron las muestras fecales de 76 niños, mediante las pruebas de esteatocrito tradicional y esteatocrito ácido, para la cuantificación de la grasa. Resultados. Se observó diarrea aguda en el 80 % de los niños y diarrea persistente en el 20 %. De los individuos con diarrea, el 40 % fue positivo para enteropatógenos; los más diagnosticados fueron rotavirus (13,3 %) y Giardia duodenalis (11,1 %). Entre los pacientes infectados, la sangre oculta fue más evidente en aquellos portadores de bacterias patógenas (40 %) o enterovirus (40%), mientras que la esteatorrea se observó en infecciones por el protozoo G. duodenalis (35,7 %). Los niños con diarrea excretaron significativamente más lípidos en las heces que aquellos sin diarrea, según lo determinado por los métodos de esteatocrito tradicional (p<0,0003) y esteatocrito ácido (p<0,0001). Conclusiones. La diarrea infantil puede provocar deficiencias graves de nutrientes. La esteatorrea es distintiva de la malabsorción intestinal y puede detectarse mediante la estimación del esteatocrito ácido. Esta prueba podría utilizarse de forma rutinaria como una herramienta de laboratorio para la evaluación semicuantitativa de la malabsorción de grasas en niños con diarrea.
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Diarrea , Heces , Síndromes de Malabsorción , Humanos , Diarrea/parasitología , Diarrea/etiología , Diarrea/microbiología , Heces/parasitología , Preescolar , Lactante , Síndromes de Malabsorción/complicaciones , Síndromes de Malabsorción/etiología , Masculino , Femenino , Niño , Giardiasis/complicaciones , Esteatorrea/etiología , Gastroenteritis/complicaciones , Gastroenteritis/parasitología , Gastroenteritis/microbiología , Sangre OcultaRESUMEN
Antimicrobials serve as crucial treatments in both veterinary and human medicine, aiding in the control and prevention of infectious diseases. However, their misuse or overuse has led to the emergence of antimicrobial resistance, posing a significant threat to public health. This review focuses on extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in animals and their associated food products, which contribute to the proliferation of antimicrobial-resistant strains. Recent research has highlighted the presence of ESBL-producing E. coli in animals and animal-derived foods, with some studies indicating genetic similarities between these isolates and those found in human infections. This underscores the urgent need to address antimicrobial resistance as a pressing public health issue. More comprehensive studies are required to understand the evolving landscape of ESBLs and to develop strategic public health policies grounded in the One Health approach, aiming to control and mitigate their prevalence effectively.
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Escherichia coli O157:H7 (E. coli O157:H7) and Campylobacter jejuni (C. jejuni) are pathogenic microorganisms that can cause severe clinical symptoms in humans and are associated with bovine meat consumption. Specific monitoring for E. coli O157: H7 or C. jejuni in meat is not mandatory under Chilean regulations. In this study, we analyzed 544 samples for the detection of both microorganisms, obtained from 272 bovine carcasses (280 kg average) at two slaughterhouses in the Bio-Bío District, Chile. Sampling was carried out at post-shower of carcasses and after channel passage through the cold chamber. Eleven samples were found to be positive for E. coli O157:H7 (4.0%) using microbiological and biochemical detection techniques and were subjected to a multiplex PCR to detect fliC and rfbE genes. Six samples (2.2%) were also found to be positive for the pathogenicity genes stx1, stx2, and eaeA. Twenty-two carcasses (8.0%) were found to be positive for C. jejuni using microbiological and biochemical detection techniques, but no sample with amplified mapA gene was found.
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Mataderos , Campylobacter jejuni , Escherichia coli O157 , Proteínas de Escherichia coli , Microbiología de Alimentos , Animales , Bovinos , Campylobacter jejuni/aislamiento & purificación , Campylobacter jejuni/genética , Escherichia coli O157/aislamiento & purificación , Escherichia coli O157/genética , Chile , Proteínas de Escherichia coli/genética , Flagelina/genética , Carne/microbiología , Contaminación de Alimentos/análisis , Adhesinas Bacterianas/genética , Toxina Shiga I/genética , Toxina Shiga II/genética , Reacción en Cadena de la Polimerasa Multiplex , Proteínas Bacterianas/genética , Transaminasas , Carbohidrato EpimerasasRESUMEN
During the COVID-19 pandemic, the occurrence of carbapenem-resistant Klebsiella pneumoniae increased in human clinical settings worldwide. Impacted by this increase, international high-risk clones harboring carbapenemase-encoding genes have been circulating in different sources, including the environment. The blaKPC gene is the most commonly disseminated carbapenemase-encoding gene worldwide, whose transmission is carried out by different mobile genetic elements. In this study, blaKPC-2-positive Klebsiella pneumoniae complex strains were isolated from different anthropogenically affected aquatic ecosystems and characterized using phenotypic, molecular, and genomic methods. K. pneumoniae complex strains exhibited multidrug-resistant and extensively drug-resistant profiles, spotlighting the resistance to carbapenems, ceftazidime-avibactam, colistin, and tigecycline, which are recognized as last-line antimicrobial treatment options. Molecular analysis showed the presence of several antimicrobial resistance, virulence, and metal tolerance genes. In-depth analysis showed that the blaKPC-2 gene was associated with three different Tn4401 isoforms (i.e., Tn4401a, Tn4401b, and Tn4401i) and NTEKPC elements. Different plasmid replicons were detected and a conjugative IncN-pST15 plasmid harboring the blaKPC-2 gene associated with Tn4401i was highlighted. K. pneumoniae complex strains belonging to international high-risk (e.g., ST11 and ST340) and unusual clones (e.g., ST323, ST526, and ST4216) previously linked to clinical settings. In this context, some clones were reported for the first time in the environmental sector. Therefore, these findings evidence the occurrence of carbapenemase-producing K. pneumoniae complex strains in aquatic ecosystems and contribute to the monitoring of carbapenem resistance worldwide.
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Antibacterianos , Variación Genética , Klebsiella pneumoniae , Pruebas de Sensibilidad Microbiana , Plásmidos , beta-Lactamasas , Humanos , Antibacterianos/farmacología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , beta-Lactamasas/genética , beta-Lactamasas/metabolismo , Carbapenémicos/farmacología , Farmacorresistencia Bacteriana Múltiple/genética , Ecosistema , Infecciones por Klebsiella/microbiología , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/aislamiento & purificación , Klebsiella pneumoniae/efectos de los fármacos , Klebsiella pneumoniae/enzimología , Plásmidos/genética , Microbiología del AguaRESUMEN
Resumen Introducción: La coloración rosa de la leche materna es poco frecuente y está asociada a colonización por Serratia marcescens. Se aísla con mayor frecuencia en entornos de cuidados intensivos. La desinformación por la coloración rosa puede conducir a una terminación prematura de la lactancia. El objetivo es presentar cuatro casos de coloración rosa de la leche materna. Métodos: Se describen dos casos retrospectivos y dos prospectivos de presentación de leche materna de color rosa. Los casos fueron reportados a la autora principal. Resultados: Se presentan cuatro binomios sanos con reporte de coloración rosa. Se aisló S. marcescens en una muestra de leche materna. Los cuatro lactantes eran asintomáticos y tuvieron cultivos para la enterobacteria. Las madres fueron tratadas con antibiótico ambulatorio. Dos lactantes recibieron tratamiento. Todos los cultivos posteriores fueron negativos y la coloración rosa cesó. Todos reanudaron la lactancia materna de forma exitosa. Conclusiones: Existen muy pocos casos de leche de color rosa reportados en la literatura mundial. La colonización por S. marcescens no es una indicación de suspensión de la lactancia.
Abstract Background: Pink coloration of breast milk is uncommon and it´s associated with colonization by Serratia marcescens, which is most frequently isolated in intensive care settings. Misinterpretation of the pink coloration may lead to premature cessation of breastfeeding. The objective is to present four cases of pink discoloration. Methods: Two retrospective and two prospective cases of pink discoloration in breast milk are described, which were reported to the lead author. Results: Four healthy mother-infant pairs with documented pink discoloration are presented. S. marcescens was isolated from breast milk samples. All four infants were asymptomatic and underwent enterobacteria cultures. The mothers received outpatient antibiotic treatment, and two infants received treatment as well. Subsequent cultures yielded negative results, and the pink discoloration ceased. All mothers successfully resumed breastfeeding. Conclusions: There are very few reported cases of pink breast milk in the global literature. Colonization by S. marcescens is not an indication for discontinuation of breastfeeding.
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The consumption of seafood is crucial for food security, but poor hygiene along the food production chain can result in low microbiological quality, posing significant risks for public health and seafood quality. Thus, this study aimed to assess the microbiological quality and antimicrobial sensitivity of E. coli from 69 samples of illegally marketed shrimp and mussels in the Vitória Region, Brazil. These foods exhibited poor microbiological quality due to high counts of mesophilic, psychrotrophic, and enterobacteria microorganisms. While this issue is widespread in this area, shrimp samples displayed higher microbial counts compared to mussels, and fresh mussels had elevated counts of enterobacteria compared to frozen ones. Among the 10 E. coli isolates, none carried the genes blaCTX-M-1, blaCTX-M-2, blaCTX-M-3, blaCTX-M-15, mcr-1, mcr-2, mcr-3, mcr-4, and tet, associated with antibiotic resistance. Phenotypical resistance to tetracycline and fosfomycin was not observed in any isolate, while only 20% demonstrated resistance to ciprofloxacin. Regarding ampicillin and amoxicillin with clavulanic acid, 60% of isolates were resistant, 10% showed intermediate susceptibility, and 30% were sensitive. One isolate was considered simultaneously resistant to ß-lactams and quinolones, and none were conserved as ESBL producers. These findings highlight the inherent risks to local public health that arise from consuming improperly prepared seafood in this area.
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AIM: The objective of this study was to investigate the antimicrobial resistance genes (ARGs) in plasmids of Enterobacteriaceae from soil, sewage, and feces of food-producing animals and humans. METHODS AND RESULTS: The plasmid sequences were obtained from the NCBI database. For the identification of ARG, comprehensive antibiotic resistance database (CARD), and ResFinder were used. Gene conservation and evolution were investigated using DnaSP v.6. The transfer potential of the plasmids was evaluated using oriTfinder and a MOB-based phylogenetic tree was reconstructed using Fastree. We identified a total of 1064 ARGs in all plasmids analyzed, conferring resistance to 15 groups of antibiotics, mostly aminoglycosides, beta-lactams, and sulfonamides. The greatest number of ARGs per plasmid was found in enterobacteria from chicken feces. Plasmids from Escherichia coli carrying multiple ARGs were found in all ecosystems. Some of the most abundant genes were shared among all ecosystems, including aph(6)-Id, aph(3'')-Ib, tet(A), and sul2. A high level of sequence conservation was found among these genes, and tet(A) and sul2 are under positive selective pressure. Approximately 62% of the plasmids carrying at least one ARG were potentially transferable. Phylogenetic analysis indicated a potential co-evolution of Enterobacteriaceae plasmids in nature. CONCLUSION: The high abundance of Enterobacteriaceae plasmids from diverse ecosystems carrying ARGs reveals their widespread distribution and importance.
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Antibacterianos , Enterobacteriaceae , Animales , Humanos , Enterobacteriaceae/genética , Antibacterianos/farmacología , Filogenia , Ecosistema , Farmacorresistencia Bacteriana/genética , Plásmidos/genética , Escherichia coli/genéticaRESUMEN
Antibiotic resistance in Citrobacter freundii is a public health concern. This study evaluated the closed genome of a C. freundii isolated from the stool of a hospitalized patient initially related to a Salmonella outbreak. Confirmation of the isolate was determined by whole-genome sequencing. Nanopore sequencing was performed using a MinION with a Flongle flow cell. Assembly using SPAdes and Unicycler yielded a closed genome annotated by National Center for Biotechnology Information Prokaryotic Genome Annotation Pipeline. Genomic analyses employed MLST 2.0, ResFinder4.1, PlasmidFinder2.1, and VFanalyzer. Phylogenetic comparison utilized the Center for Food Safety and Applied Nutrition (CFSAN)-single nucleotide polymorphism pipeline and Genetic Algorithm for Rapid Likelihood Inference. Antimicrobial susceptibility was tested by broth microdilution following Clinical and Laboratory Standards Institute criteria. Multi-locus sequence type in silico analysis assigned the C. freundii as sequence type 64 and the blaCMY-41 gene was detected in resistome investigation. The susceptibility to antibiotics, determined using Sensititre® plates, revealed resistance to aztreonam, colistin, cefoxitin, amoxicillin/clavulanic acid, sulfisoxazole, ampicillin, and streptomycin. The genetic relatedness of the C. freundii CFSAN077772 with publicly available C. freundii genomes revealed a close relationship to a C. freundii SRR1186659, isolated in 2009 from human stool in Tanzania. In addition, C. freundii CFSAN077772 is nested in the same cluster with C. freundii clinical strains isolated in Denmark, Mexico, Myanmar, and Canada, suggesting a successful intercontinental spread.
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Citrobacter freundii , Infecciones por Enterobacteriaceae , Humanos , Citrobacter freundii/genética , beta-Lactamasas/genética , Tipificación de Secuencias Multilocus , Filogenia , Infecciones por Enterobacteriaceae/epidemiología , Antibacterianos/farmacología , Genómica , Pruebas de Sensibilidad MicrobianaRESUMEN
Abstract Introduction. The multifactorial etiology of gastroenteritis emphasizes the need for different laboratory methods to identify or exclude infectious agents and evaluate the severity of diarrheal disease. Objective. To diagnose the infectious etiology in diarrheic children and to evaluate some fecal markers associated with intestinal integrity. Materials and methods. The study group comprised 45 children with diarrheal disease, tested for enteropathogens and malabsorption markers, and 76 children whose feces were used for fat evaluation by the traditional and acid steatocrit tests. Results. We observed acute diarrhea in 80% of the children and persistent diarrhea in 20%. Of the diarrheic individuals analyzed, 40% were positive for enteropathogens, with rotavirus (13.3%) and Giardia duodenalis (11.1%) the most frequently diagnosed. Among the infected patients, occult blood was more evident in those carrying pathogenic bacteria (40%) and enteroviruses (40%), while steatorrhea was observed in infections by the protozoa G. duodenalis (35.7%). Children with diarrhea excreted significantly more lipids in feces than non-diarrheic children, as determined by the traditional (p<0.0003) and acid steatocrit (p<0.0001) methods. Moreover, the acid steatocrit method detected 16.7% more fecal fat than the traditional method. Conclusions. Childhood diarrhea can lead to increasingly severe nutrient deficiencies. Steatorrhea is the hallmark of malabsorption, and a stool test, such as the acid steatocrit, can be routinely used as a laboratory tool for the semi-quantitative evaluation of fat malabsorption in diarrheic children.
Resumen Introducción. La etiología multifactorial de la gastroenteritis enfatiza la necesidad de usar diferentes métodos de laboratorio para identificar o excluir agentes infecciosos y evaluar la gravedad de la enfermedad diarreica. Objetivo. Diagnosticar la etiología infecciosa de la diarrea en niños y evaluar algunos marcadores fecales asociados con la integridad intestinal. Materiales y métodos. Se estudiaron 45 niños con enfermedad diarreica, en los cuales se evaluaron la presencia de enteropatógenos y los marcadores de malabsorción. Se analizaron las muestras fecales de 76 niños, mediante las pruebas de esteatocrito tradicional y esteatocrito ácido, para la cuantificación de la grasa. Resultados. Se observó diarrea aguda en el 80 % de los niños y diarrea persistente en el 20 %. De los individuos con diarrea, el 40 % fue positivo para enteropatógenos; los más diagnosticados fueron rotavirus (13,3 %) y Giardia duodenalis (11,1 %). Entre los pacientes infectados, la sangre oculta fue más evidente en aquellos portadores de bacterias patógenas (40 %) o enterovirus (40%), mientras que la esteatorrea se observó en infecciones por el protozoo G. duodenalis (35,7 %). Los niños con diarrea excretaron significativamente más lípidos en las heces que aquellos sin diarrea, según lo determinado por los métodos de esteatocrito tradicional (p<0,0003) y esteatocrito ácido (p<0,0001). Conclusiones. La diarrea infantil puede provocar deficiencias graves de nutrientes. La esteatorrea es distintiva de la malabsorción intestinal y puede detectarse mediante la estimación del esteatocrito ácido. Esta prueba podría utilizarse de forma rutinaria como una herramienta de laboratorio para la evaluación semicuantitativa de la malabsorción de grasas en niños con diarrea.
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AIMS: To evaluate the composition and functions of the gut microbiota in patients with decompensated alcohol-associated cirrhosis, with and without hepatic encephalopathy (HE). METHODS AND RESULTS: Faecal samples from 31 inpatients (20 with HE, 11 without HE), and from 18 age-balanced healthy controls (HC), were included. Microbial composition was determined by 16S rRNA amplicon sequencing and analysed using QIIME2. Metabolic pathways were inferred by PICRUSt2, and short-chain fatty acids (SCFAs) quantification was performed by gas chromatography. The gut microbiota of patients with HE was characterized by a diminished α-diversity, compared to no-HE (P < 0.01) and HC (P < 0.001) groups; ß-diversity also differed between HE vs no-HE patients (P < 0.05), and between HE vs HC (P < 0.001). In patients with HE, Escherichia/Shigella, Burkholderiales and Lactobacillales taxa predominated. In contrast, patients without HE were characterized by Veillonella and Bacteroides. Reduced levels of faecal SCFAs in both groups correlated with a depletion of beneficial taxa, such as Ruminococcus or Faecalibacterium. PICRUSt2 analysis showed both an enhanced catabolism of arginine through ammonia-producing pathways and chorismate biosynthesis in HE patients, a key precursor of aromatic amino acids. CONCLUSIONS: The gut microbiota of HE patients exhibits a proinflammatory dysbiotic profile, plus metabolic pathways that produce potentially neurotoxic byproducts.
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Microbioma Gastrointestinal , Encefalopatía Hepática , Microbiota , Humanos , Encefalopatía Hepática/microbiología , Arginina , ARN Ribosómico 16S/genética , Heces/microbiología , Ácidos Grasos Volátiles/análisisRESUMEN
Little is known about the influence of the urban environments on bat species 'ecology. The urbanization process potentially lead to critical ecological changes in bat communities' intra and interspecific pathogenic transmissions dynamics. To date, the monitoring of pathogens in bats in Brazil has only been done with bats found dead or alive in households, from rabies surveillance systems. The present work aimed to investigate how urbanization influenced bat richness, relative abundance and pathogen occurrence. Most captured bats were Phyllostomidae, especially Sturnira lilium, Artibeus lituratus, A. fimbriatus, Glossophaga soricina, and Platyrrhinus lineatus, among others. From preserved-rural towards urban areas the lesser the bat richness, the higher the relative abundance of the captured bats. Noise level, luminosity and relative humidity correlated with bat abundance. The proportion of genders, sexually active bats and their size (weight, right forearm length, and body condition index) were stable throughout the investigation. Still, the proportion of pregnant females was higher in Spring and the number of juveniles in Summer, evidencing the seasonality of reproduction. Several Enterobacteria were isolated, evidencing a significant role of bats in the circulation of pathogens of medical and veterinary interest. These results are crucial in the pursuit of a harmonious coexistence between humans, bats and domestic animals in areas with different levels of anthropization.
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Quirópteros , Rabia , Humanos , Femenino , Animales , Masculino , Brasil/epidemiología , Rabia/epidemiología , Rabia/prevención & control , Rabia/veterinaria , Animales Domésticos , ReproducciónRESUMEN
The aim of this work was to evaluate the anti-Escherichia coli effect of cell-free supernatant (CFS) of Lactobacillus spp. against planktonic and biofilm forms of foodborne isolates. Escherichiacoli strains (P12, P25, P35 and P36), previously isolated from fresh filets of fish, were subjected to antimicrobial susceptibility determination by the disc-diffusion agar method. Subsequently, the antagonistic effect between probiotic and pathogenic strains was determined by spot overlay assay. Finally, the CFS activity against pre-established (12 h) biofilms was demonstrated through biomass quantification by crystal violet staining and scanning electron microscopy (SEM). All isolates presented some pattern of resistance, primarily to ampicillin and tetracycline. Probiotic strains presented high antagonistic effects against all E. coli strains, presenting inhibition zones (R) ranging from 15.60 to 20.67 mm. Additionally, the residual biomass of pre-established (12 h) biofilm was drastically reduced about 50% after CFS treatment (P < 0.01). What can be noted by SEM images, which show less surface-attached cells of CFS-treated biofilms of E. coli (P12). Thus, cell-free preparations produced from Lactobacillus spp. may represent a tool in the battle against planktonic cells and biofilm forms of antibiotic-resistant E. coli.
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Escherichia coli , Animales , Lactobacillus , Biopelículas , Antibacterianos/farmacologíaRESUMEN
Bovine mastitis is a complex disease that brings great losses to the dairy producer. The microbial diversity of the soils, as well as the presence of resistance genes in the environment directly influence the maintenance of mastitis in the farm. The objective of this work was to analyze the bacterial diversity in pasture soils of a dairy family farm, detecting enterobacteria that may be involved in the etiology of bovine mastitis, and to detect genes that encode broad-spectrum betalactamases in these soils. Twelve soil samples, representative of different areas of the farm located in the municipality of Barra do Piraí, Rio de Janeiro, were collected at different times of the year. Total DNA was extracted from the samples, gene amplified by Nested-PCR and then the amplification products were separated by DGGE (Denaturing Gradient Gel Electrophoresis). With the DGGE it was possible to construct dendograms that effectively represented the bacterial diversity of these soils. Eight of the soil samples were used to amplify the genes encoding the betalactamase enzymes TEM (blaTEM gene), SHV (blaSHV gene) and CTX (blaCTXM gene). In three of the eight soil samples, the blaSHV gene was found to be present. The blaTEM and blaCTX-M genes were not detected in any of the samples. The detection of genes encoding broad-spectrum betalactamases in dairy cattle pasture soils is of concern, because the transfer of gene material between pathogenic and non-pathogenic bacteria in this environment is a reality
A mastite bovina é uma doença complexa que traz grandes prejuízos ao produtor de leite. A diversidade micro-biana dos solos, bem como a presença de genes de resistência no ambiente, influencia diretamente a manutenção da mastite na fazenda. O objetivo deste trabalho foi analisar a diversidade bacteriana em solos de pastagem de uma propriedade familiar leiteira, detectando enterobactérias que possam estar envolvidas na etiologia da mastite bovina, e detectar genes que codificam betalactamases de amplo espectro nesses solos. Doze amostras de solo, representativas de diferentes áreas da fazenda localizada no município de Barra do Piraí, Rio de Janeiro, foram coletadas em diferentes épocas do ano. O DNA total foi extraído das amostras, o gene foi amplificado por Nested-PCR e, em seguida, os produtos de amplificação foram separados por DGGE (Denaturing Gradient Gel Electrophoresis). Com a DGGE, foi possível construir dendogramas que representavam efetivamente a diversidade bacteriana desses solos. Oito das amostras de solo foram usadas para amplificar os genes que codificam as enzimas betalactamase TEM (gene blaTEM), SHV (gene blaSHV) e CTX (gene blaCTXM). Em três das oito amostras de solo, foi constatada a presença do gene blaSHV. Os genes blaTEM e blaCTX-M não foram detectados em nenhuma das amostras. A detecção de genes que codificam betalactamases de amplo espectro em solos de pastagens de gado leiteiro é preocupante, pois a transferência de material genético entre bactérias patogênicas e não patogênicas nesse ambiente é uma realidade
Asunto(s)
Microbiología del Suelo , beta-Lactamasas/análisis , Pastizales , Enterobacteriaceae/aislamiento & purificación , Electroforesis en Gel de Gradiente Desnaturalizante/veterinariaRESUMEN
Information regarding resistance and virulence traits of meningitis-causing enterobacteria in hospital environment remains scarce. The aim of this study was to characterize virulence and acquired resistance genes of carbapenem-resistant and/or 3rd to 4th generation cephalosporin-resistant Klebsiella pneumoniae isolated from the cerebrospinal fluid of inpatients. Antimicrobial susceptibility testing was performed by disk diffusion. The string test was performed to identify hypermucoviscous phenotype. Galleria mellonella infection model was used to evaluate the virulence profile of the isolates. Screening for virulence determinants and acquired resistance genes were performed by PCR. The blaCTX-M and/or blaKPC and/or rmtG were detected in all the isolates. Genetic virulence determinants, including mrkD, entB, iroD, fecIRA, uge, wabG, fimH, ureA, ybtS, and clb were detected in the majority of multidrug-resistant K. pneumoniae isolates. One isolate presented hypermucoviscous phenotype, and several isolates showed enhanced virulence in G. mellonella infection model. The combination of the virulence genes found here seems to support not only the known virulence genetic context among nosocomial infections-causing K. pneumoniae but also the role that clb and ybtS may play in K. pneumoniae virulence.
Asunto(s)
Infecciones por Klebsiella , Klebsiella pneumoniae , Antibacterianos/farmacología , Carbapenémicos , Cefalosporinas , Humanos , Infecciones por Klebsiella/microbiología , Pruebas de Sensibilidad Microbiana , Urea , Virulencia/genética , Factores de Virulencia/genética , beta-Lactamasas/genéticaRESUMEN
The reference material (RM) is a technical requirement for the quality assurance of analytical results and proficiency tests or interlaboratory comparisons. Microbiological RMs are most available in the dehydrated form, mainly by freeze-drying, and maintaining bacterial survival after preparation is a challenge. Thus, obtaining the most resistant cells is essential. Considering that bacteria present cross-response to dehydration after being submitted to an array of stress conditions, this study aimed to evaluate the influence of growth conditions on enterobacteria for the production of mixed microbiological RMs by freeze-drying in skim milk powder. Salmonella enterica serovar Enteritidis, Cronobacter sakazakii, Escherichia coli, and Citrobacter freundii were grown in a minimal medium with 0.5 M NaCl and 0 to 5.0 mM of manganese sulfate (MnSO4) until stationary phase. Salmonella Enteritidis presented an increased resistance to dehydration in the presence of Mn, while C. sakazakii was the most resistant to freeze-drying and further storage for 90 days. Mixed microbiological RMs were produced by freeze-drying and containing Salmonella Enteritidis and coliforms in skim milk powder with 100 mM of trehalose and the Salmonella survival rate was 91.2 to 93.6%. The mixed RM was stable after 30 days at -20 °C, and Salmonella and coliforms were detected by different methods being, the Rambach Agar the best for the bacterial differentiation. The results showed that the culture conditions applied in this study resulted in bacterial cells being more resistant to dehydration, freeze-drying, and stabilization for the production of mixed microbiological RMs more stable and homogeneous.
Asunto(s)
Deshidratación , Salmonella , Humanos , Viabilidad Microbiana , Polvos , Liofilización/métodos , Bacterias , Microbiología de AlimentosRESUMEN
Surgical environment can play as a source of multidrug-resistance organism, what can pose as a big threat to the patients and health care professionals. This study aimed to evaluate the prevalence and antimicrobial resistance profile of Gram-positive cocci (GPC) and Gram-negative bacilli (GNB) isolated from the surgical environment. All samples were collected during the intraoperative period of clean/clean-contaminated (G1) and contaminated (G2) surgery. A total of 150 samples were collected from the superficial surgical site in the beginning (nâ¯=â¯30) and the end (nâ¯=â¯30) of the procedure, surgeon's hands before (nâ¯=â¯30) and after (nâ¯=â¯30) antisepsis, and the surgical environment (nâ¯=â¯30). MALDI-TOF MS and antimicrobial susceptibility testing by disk diffusion method were performed for species identification, and determination of the resistance profile. Sixty-eight isolates of GPC and 15 of GNB were obtained. Staphylococcus spp. were the most frequent species isolated from surgical site (55.26% [21/38]), surgeon's hands (46.15% [6/13]), and environment (56.67% [17/30]). GPC were mostly resistance to penicillin (85.71% [54/63]), and erythromycin (77.78% [49/63]), and GNB were mostly resistance to cefazolin (58.33% [7/12]), and azithromycin (58.33% [7/12]). High incidence of multidrug resistance was observed in coagulase-negative staphylococci (86.21% [25/29]), coagulase-positive staphylococci (86.67% [13/15]), Enterococcus spp. (68.42% [13/19]) and Gram-negative bacilli (60% [9/15]). The high rate of resistance of commensal bacteria found in our study is worrying. Coagulase-negative staphylococci are community pathogens related to nosocomial infections in human and veterinary hospitals, their presence in healthy patients and in veterinary professionals represent an important source of infection in the One Health context. Continuous surveillance and application of antimicrobial stewardship programs are essential in the fight against this threat.
Asunto(s)
Bacterias Gramnegativas , Cocos Grampositivos , Mano , Hospitales Veterinarios , Quirófanos , Cirujanos , Animales , Antibacterianos/farmacología , Bacterias , Brasil , Coagulasa , Perros , Farmacorresistencia Bacteriana Múltiple , Bacterias Gramnegativas/aislamiento & purificación , Cocos Grampositivos/aislamiento & purificación , Mano/microbiología , Hospitales de Enseñanza , Humanos , Pruebas de Sensibilidad Microbiana/veterinaria , StaphylococcusRESUMEN
Emergency department areas were repurposed as intensive care units (ICUs) for patients with acute respiratory distress syndrome during the initial months of the coronavirus disease 2019 (COVID-19) pandemic. We describe an outbreak of New Delhi metallo-ß-lactamase 1 (NDM-1)-producing Escherichia coli infections in critically ill COVID-19 patients admitted to one of the repurposed units. Seven patients developed infections (6 ventilator-associated pneumonia [VAP] and 1 urinary tract infection [UTI]) due to carbapenem-resistant E. coli, and only two survived. Five of the affected patients and four additional patients had rectal carriage of carbapenem-resistant E. coli. The E. coli strain from the affected patients corresponded to a single sequence type. Rectal screening identified isolates of two other sequence types bearing blaNDM-1. Isolates of all three sequence types harbored an IncFII plasmid. The plasmid was confirmed to carry blaNDM-1 through conjugation. An outbreak of clonal NDM-1-producing E. coli isolates and subsequent dissemination of NDM-1 through mobile elements to other E. coli strains occurred after hospital conversion during the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. This emphasizes the need for infection control practices in surge scenarios. IMPORTANCE The SARS-CoV-2 pandemic has resulted in a surge of critically ill patients. Hospitals have had to adapt to the demand by repurposing areas as intensive care units. This has resulted in high workload and disruption of usual hospital workflows. Surge capacity guidelines and pandemic response plans do not contemplate how to limit collateral damage from issues like hospital-acquired infections. It is vital to ensure quality of care in surge scenarios.
Asunto(s)
Infección Hospitalaria/microbiología , Infecciones por Escherichia coli/microbiología , Escherichia coli/enzimología , Escherichia coli/aislamiento & purificación , beta-Lactamasas/metabolismo , Adulto , Anciano , COVID-19/epidemiología , COVID-19/virología , Conjugación Genética , Infección Hospitalaria/epidemiología , Brotes de Enfermedades , Escherichia coli/clasificación , Escherichia coli/genética , Infecciones por Escherichia coli/epidemiología , Infecciones por Escherichia coli/mortalidad , Femenino , Humanos , Unidades de Cuidados Intensivos/estadística & datos numéricos , Masculino , México/epidemiología , Persona de Mediana Edad , Plásmidos/genética , SARS-CoV-2/fisiología , Centros de Atención Terciaria/estadística & datos numéricos , beta-Lactamasas/genéticaRESUMEN
Escherichia coli is a pathogen associated with infections in piglets in the post-weaning phase, its pathogenicity is related to the animal's susceptibility to bacterial enterotoxins. The objective of the present study was to determine the EOs activity against E. colistrain, in the form planktonic and sessile. Although the Disc-Diffusion tests to determine the Minimum Inhibitory Concentration, do not fully corroborate with the other analyzes of this study, it was noticed bacteria inhibition. The EOs were prepared at 0.4%, 0.8% and 1.0% for tests. The tested EOs were effective against E. coliplanktonic cells (p<0.05). As for the sessile cells, the most significant result was inhibition and 100% sessile cells at the concentration of 1.0% of Cymbopogon citratusEO. Although there was resistance in some treatments, the tested EOs demonstrated inhibition capacity, constituting promising alternatives for the control of E. coli, especially of planktonic cells.
Escherichia coli es un patógeno asociado con infecciones en lechones en la fase posterior al destete, su patogenicidad está relacionada con la susceptibilidad del animal a las enterotoxinas bacterianas. El objetivo del presente estudio fue determinar la actividad de contra E. coli, en la forma planctónico y sésil. Aunque las pruebas de difusión de disco para determinar la concentración inhibitoria mínima, no corroboran completamente con los otros análisis de este estudio, se observó inhibición de la bacteria. Las soluciones basadas en AE se prepararon al 0.4%, 0.8% y 1.0% para pruebas. Los AEs probados fueron efectivos contra las células planctónicas (p<0.05). En cuanto a las células sésiles, el resultado más significativo fue la inhibición y el 100% de las células sésiles a la concentración de 1,0% de Cymbopogon citratus. Aunque hubo resistencia en algunos tratamientos, los AEs probados demostraron capacidad de inhibición, constituyendo alternativas prometedoras para el control de E. coli, especialmente de células planctónicas.