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The growing number of sequenced genomes enables the study of secondary metabolite biosynthetic gene clusters (BGC) in phyla beyond well-studied soil bacteria. We mined 2627 enterobacterial genomes to detect 8604 BGCs, including nonribosomal peptide synthetases, siderophores, polyketide-nonribosomal peptide hybrids, and 60 other BGC types, with an average of around 3.3 BGCs per genome. These BGCs represented 212 distinct BGC families, of which only 20 have associated products in the MIBiG standard database with functions such as siderophores, antibiotics, and genotoxins. Pangenome analysis identified genes associated with a specific BGC encoding for colon cancer-related colibactin. In one example, we associated genes involved in the type VI secretion system with the presence of a colibactin BGC inEscherichia. This richness of BGCs in enterobacteria opens up the possibility to discover novel secondary metabolites, their physiological roles and provides a guide to identify and understand PKS associated gene sets.

Background and Aim: There is a worldwide controversy about the choice of microbial flora for use as process hygiene indicators. This study aimed to evaluate the pertinence of using either coliforms or Enterobacteriaceae (EB) as process hygiene indicators in the pasteurized milk production line. Two flora families and total flora were used as bacterial indicators in some stages of pasteurized milk production line to identify the origin of post-pasteurization contamination and compare the results obtained for each flora. In addition, the bacteriological profile of isolated coliforms and EB was developed. Materials and Methods: One thousand and two hundred samples of pasteurized cow milk and surfaces (pipes and tank) at various processing stages were taken from two dairies in the northern region of Algeria. The total microbial flora (TF), total coliforms (TC), thermotolerant coliforms, and EB were enumerated, following the recommendations of ISO 4833:2006, ISO 4832:2006, and ISO 21528-2:2017 methods, respectively. The bacteriological profile was determined using the API 20E and 10S tests (bioMérieux, France). Furthermore, the cleaning efficiency and disinfection protocol of surfaces were evaluated using contact agar slides 1 (Liofilchem™, Italy). Results: Enumeration of the different indicators shows that the highest contamination rate is recorded by the total flora in the two units, 3.28 and 3.78 log CFU/mL, respectively. EB (–0.60 log CFU/mL) at post-pasteurization stage in Unit 1 and coliforms (0.44 log CFU/mL) at the pasteurized packaged milk stage in Unit 2 are the least significant germ families. The lowest compliance rates of bacterial contamination were reported for total flora (82-85%) at the three sampled sites in Unit 2. In comparison, the highest was reported in Unit 1 (99.8%) and 2 (98%) by the EB indicator. Assessing the surface cleaning and disinfection protocol compliance shows that the tank records the highest non-compliance rates for EB and TF (4% and 3%) in Unit 2. EB are represented in both units by various species. Acinetobacter baumannii in Unit 1 and Enterobacter cloacae in Unit 2 are the common species of the three indicator families. Acinetobacter and Enterobacter in Unit 1, Escherichia, Citrobacter, Enterobacter, Klebsiella, and Hafnia in Unit 2 are the most time persistent bacterial genera along the production line. Stenotrophomonas, Serratia, Salmonella, Enterobacter, and Escherichia are common genera in both units. Conclusion: The results obtained show no difference in the use of EB or TC as hygiene indicators. However, if the objective is to identify the species of bacterial populations, using EBs are the most appropriate.

The grey-breasted parakeet (Pyrrhura griseipectus) is an endangered psittacine species that have been affected by illegal trade and deforestation. Currently, this endemic species is only found in three areas in Ceará state, in Brazil. This study aimed to investigate the frequency and diversity of Enterobacteriaceae in wild adult grey-breasted parakeets and determine their susceptibility to antimicrobial agents. Cloacal swab samples were collected from 27 individuals and environmental swabs (drag swabs) from five nests used by these birds. Twenty-seven strains from nine species of Enterobacteriaceae were recovered from cloacal swabs, and the most prevalent bacteria strains were Hafnia alvei (22%) and Pantoea agglomerans (22%). From environmental nest samples, seven strains from three bacterial species were isolated, being the P. agglomerans the most frequent species (100%). Twenty-two of the 27 isolates (81.4%) exhibited antibiotic resistance, varying from one to eight of the 12 antimicrobials commonly used. Resistance to amoxicillin was the most prevalent (70.4%), followed by azithromycin (22.2%) and ceftriaxone (18.5%). None of the strains were resistant to gentamicin, tobramycin, ciprofloxacin or tetracycline. The H. alvei was the main species presenting multidrug resistance, including resistance against meropenem, which is an important finding. These results could provide interesting information on the health of these endangered wild grey-breasted parakeets. They could also indicate that the obtained isolates are part of a group of bacteria that are typical components of the enteric microbiota of birds, which present elevated rates of resistance to amoxicillin. RESUMO: O periquito-de-cara-suja (Pyrrhura griseipectus) é uma espécie de psitacídeo considerado pela IUCN como ameaçado de extinção, resultado do comércio ilegal e do desmatamento. Atualmente, essa espécie endêmica é encontrada apenas em três áreas no estado do Ceará, Brasil. O objetivo deste estudo foi investigar a frequência e a diversidade de Enterobacteriaceae em periquitos de peito cinza adultos selvagens e determinar sua suscetibilidade a agentes antimicrobianos. Amostras de suabes cloacais foram coletadas de 27 indivíduos e de suabes ambientais (suabes de arrasto) de cinco ninhos utilizados por essas aves. Vinte e sete cepas de nove espécies de Enterobacteriaceae foram isoladas a partir de suabes cloacais, sendo as cepas bacterianas mais prevalentes Hafnia alvei (22%) e Pantoea agglomerans (22%). Das amostras ambientais de ninhos foram isoladas sete linhagens de três espécies bacterianas, sendo P. agglomerans a espécie mais frequente (100%). Vinte e dois dos 27 isolados (81,4%) exibiram resistência a antibióticos, variando de um a oito dos 12 antimicrobianos comumente usados. A resistência a amoxicilina foi a mais prevalente (70,4%), seguida por azitromicina (22,2%) e ceftriaxona (18,5%). Nenhuma das cepas era resistente à gentamicina, tobramicina, ciprofloxacina ou tetraciclina. H. alvei foi a principal espécie que apresentou resistência a múltiplas drogas e que também esteve associada a um outro achado relevante desta pesquisa, que foi a detecção de um caso de resistência ao meropenem. Esses dados fornecem informações relevantes sobre a saúde desses periquitos selvagens ameaçados e permite concluir que os isolados obtidos fazem parte de um grupo de bactérias que normalmente compõe a microbiota entérica das aves, sendo a amoxicilina envolvida em elevadas taxas de resistência.

Background Antimicrobial combinations have been proven as a promising approach in the confrontation with multi-drug resistant bacterial pathogens. In the present study, we identify and characterize a synergistic interaction of broad-spectrum nitroreductase-activated prodrugs 5-nitrofurans, with a secondary bile salt, sodium deoxycholate (DOC) in growth inhibition and killing of enterobacteria. Results Using checkerboard assay, we show that combination of nitrofuran furazolidone (FZ) and DOC generates a profound synergistic effect on growth inhibition in several enterobacterial species including Escherichia coli, Salmonella enterica, Citrobacter gillenii and Klebsiella pneumoniae. The Fractional Inhibitory Concentration Index (FICI) for DOC-FZ synergy ranges from 0.125 to 0.35 that remains unchanged in an ampicillin-resistant E. coli strain containing a β-lactamase-producing plasmid. Findings from the time-kill assay further highlight the synergy with respect to bacterial killing in E. coli and Salmonella. We further characterize the mechanism of synergy in E. coli K12, showing that disruption of the tolC or acrA genes that encode components of multidrug efflux pumps causes, respectively, a complete or partial loss, of the DOC-FZ synergy. This finding indicates the key role of TolC-associated efflux pumps in the DOC-FZ synergy. Overexpression of nitric oxide-detoxifying enzyme Hmp results in a three-fold increase in FICI for DOC-FZ interaction, suggesting a role of nitric oxide in the synergy. We further demonstrate that DOC-FZ synergy is largely independent of NfsA and NfsB, the two major activation enzymes of the nitrofuran prodrugs. Conclusions This study is to our knowledge the first report of nitrofuran-deoxycholate synergy against Gram-negative bacteria, offering potential applications in antimicrobial therapeutics. The mechanism of DOC-FZ synergy involves FZ-mediated inhibition of TolC-associated efflux pumps that normally remove DOC from bacterial cells. One possible route contributing to that effect is via FZ-mediated nitric oxide production.

Aim. The purpose was to assess the species composition, virulence factors and the nature of the interaction of Enterobacteria with intestinal microsymbionts in HIV-positive children.Material and methods. The composition of intestinal microbiocenosis of 89 HIV-infected children (main group) and 74 children without HIV status (comparison group) was studied. 273 strains of Enterobacteria were isolated and identified to the species. The frequency and level of expression of adhesion, invasion, toxin formation and antagonism factors were studied.Results. In HIV-positive children colonization rate of intestinal mucosa E. coli lac — was 5 times higher, E. coli hly+ 1.5 times higher than in the comparison group. The quantitative level of opportunistic enterobacteria increases with the aggravation of microecological disorders, 98% of enterobacteria forms a many-to-many association. Of these, 86.8% are neutral towards other symbionts. The frequency and level of production of adhesion, invasion and toxin formation factors did not differ in the compared groups.Conclusion. In HIV-infected children is formed enterobacteriales type microbiota. Biological properties of enterobacteria did not depend on the presence of HIV status in children.

The aim of the present study was to characterize extended-spectrum β-lactamase (ESBLs) genes in multidrug resistant enterobacterial pathogens as well as commensal isolates from the Sudan during the period 2003 to 2007. ESBL production was determined phenotypically by the combined disc method, and was characterized genotypically by the detection of blagenes by PCR and nucleotide sequencing. Transferability was examined by conjugation with nalidixic-acid resistant E. coli K12. The results showed that a total of 106 of the 113 (94%) isolates including E. coli, Klebsiella pneumoniae, proteus spp., Enterobacter cloacae, Providencia spp. and Morganella morganii, were positive for blagenes including the prototype blaTEM. Eleven isolates (28%) of the 113 were ESBL producers encoding blaSHV genes (SHV5, SHV5a, SHV12, SHV26, SHV28 and SHV38), 90 isolates (80%) were CTX-M positive. All, but only one (CTX-M9) were CTX-M15. Only 3(2.7%) of the isolates were Amp-C producers (CMY-4 and DHA-1). Plasmid transfer of the multiple resistance patterns was achieved among all the isolates. These findings demonstrated that ESBLs were highly produced by multi-resistant enterobacterial isolates from the Sudan; among both clinical pathogens as well as stool commensals. This is the first report of ESBLs genes characterization from the Sudan.

Introduction. There is an urgent need for effective therapies against bacterial infections, especially those caused by antibiotic-resistant Gram-negative pathogens. Hypothesis. Synergistic combinations of existing antimicrobials show promise due to their enhanced efficacies and reduced dosages which can mitigate adverse effects, and therefore can be used as potential antibacterial therapy. Aim. In this study, we sought to characterize the in vitro interaction of 5-nitrofurans, vancomycin and sodium deoxycholate (NVD) against pathogenic bacteria. Methodology. The synergy of the NVD combination was investigated in terms of growth inhibition and bacterial killing using checkerboard and time-kill assays, respectively. Results. Using a three-dimensional checkerboard assay, we showed that 5-nitrofurans, sodium deoxycholate and vancomycin interact synergistically in the growth inhibition of 15 out of 20 Gram-negative strains tested, including clinically significant pathogens such as carbapenemase-producing Escherichia coli , Klebsiella pneumoniae and Acinetobacter baumannii , and interact indifferently against the Gram-positive strains tested. The time-kill assay further confirmed that the triple combination was bactericidal in a synergistic manner. Conclusion. This study demonstrates the synergistic effect of 5-nitrofurans, sodium deoxycholate and vancomycin against Gram-negative pathogens and highlights the potential of the combination as a treatment for Gram-negative and Gram-positive infections.

Microbiological studies of the sanitary and health status of psittacine birds that will be reintroduced is important in evaluating whether these animals act as carriers of pathogenic agents to other animals and humans. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a faster and more accurate method to identify bacteria than conventional microbiology methods. The aim of this study was to evaluate the health status of psittacines housed in captivity, by assessment of Gram-negative bacteria from fecal microbiota through MALDI- TOF MS identification. The results indicate high frequency of Gram-negative bacteria in feces (96.5%), especially from the Enterobacteriaceae family (88.7%). The most prevalent bacteria were Escherichia coli (39.0%), Proteus vulgaris (12.2%), Klebsiella spp. (12.1%) and Raoultella ornithinolytica (8.7%). Proteus hauseri, Citrobacter spp., Morganella morgannii, Providencia rettgeri, Enterobacter spp. and Escherichia hermannii were isolated with lower frequency. . All these agents are potentially pathogenic for parrots and can cause systemic infections in other animals and humans. These findings reinforce that MALDI- TOF MS proved to be a rapid and accurate method of identification of the microorganism and evaluation of the health status of psittacines, providing relevant data to assist decision-making regarding the sanitary protocols in wildlife centers, and possible future reintroduction of wild birds. Resumo Estudos microbiológicos da sanidade de psitacídeos que serão reintroduzidos são importantes para avaliar se esses animais atuam como portadores de agentes patogênicos para outros animais e humanos. A espectrometria de massa por ionização/dessorção de matriz assistida por laser/tempo de vôo (MALDI-TOF MS) é um método mais rápido e preciso para identificar bactérias na comparação com métodos convencionais de microbiologia. O objetivo deste estudo foi avaliar o estado de saúde de psitacídeos cativos, identificando bactérias Gram-negativas da microbiota fecal por MALDI -TOF MS. Os resultados indicaram alta frequência de bactérias Gram-negativas nas fezes (96,5%), principalmente da família Enterobacteriaceae (88,7%). As mais prevalentes foram Escherichia coli (39,0%), Proteus vulgaris (12,2%), Klebsiella spp. (12,1%) e Raoultella ornithinolytica (8,7%). Proteus hauseri, Citrobacter spp., Morganella morgannii, Providencia rettgeri, Enterobacter spp. e Escherichia hermannii foram isolados com menor frequência. Todos esses agentes são potencialmente patogênicos para os papagaios e podem causar infecções sistêmicas em outros animais e seres humanos. Esses achados reforçam que o MALDI- TOF MS é um método rápido e preciso de identificação do microrganismo e avaliação do estado de saúde dos psitacídeos, fornecendo dados relevantes para auxiliar na tomada de decisões sobre os protocolos sanitários em centros de triagem de animais selvagens e sobre a possibilidade de reintrodução futura.

INTRODUCTION: Inadequate wastewater treatment and fecal contamination have a strong environmental impact on antimicrobial resistance (AMR). This study evaluated the profile of AMR enterobacteria and fecal contamination from four surface waters: Jiquiriça-Brejões River and Cabrito, Tororó, and Abaeté Lagoons. METHODS: We analyzed AMR β-lactamase genes using the polymerase chain reaction method and fecal contamination using Coliscan®. RESULTS: We found high levels of fecal contamination, β-lactamase producers, and AMR genes (blaOXA-48, blaSPM, and blaVIM) in all waterbodies. CONCLUSIONS: Poor sanitation evidenced by fecal contamination and human activities around these surface waters contributed to the distribution and increase in AMR enterobacteria.

88 samples of traditional and industrial table olives were collected from various outlets in the city of Tetouan. The physical-chemical controls concerned: pH , ORP (oxydo-reducing potential) and TDE (total dry extract). The microbiological side are: total aerobic mesophilic flora ( TAMB), total coliforms (TC), Staphylococcus aureus (SA) and lactic bacteria (LB) with an identification of the most predominant enterobacteria species and their sensitivity. Also to investigate the antimicrobial activity of Olive (Olea europaea L.) .The obtained results for traditional olives far exceed food hygiene standards, especially when 29.20% of traditional samples are too loaded in TAMB and TC and 16.5% are contaminated with SA. This shows that good hygiene practices are not respected. As for industrial olives, the averages in TAMB and SA are below the thresholds set by the standards, and only 19.8% are contaminated with TC which indicates the compliance of the majority of industrial type. Black table olives are the most contaminated. The identification results of coliforms extracted from traditional olives show the predominance of Citrobacter freundii and klebsiella pneumoniae .The sensitivity test shows that 80% of Enterobacteriaceae tested were sensitive to all antibiotics.The result showed that the ethanolic extrats was active against all bacteria.

Infections of genotoxic Escherichia coli spread concomitantly with urbanized progression. These bacteria may prompt cell senescence and affect DNA stability, inducing cancer via the production of colibactin, a genotoxin shown capable of affecting host DNA in eukaryotic cells. In this study, we show that the action of colibactin may also be directed against other bacteria of the gut microbiota in which genotoxic E. coli bacteria have been introduced. Indeed, the presence of genotoxic E. coli induced a change in both the structure and function of the gut microbiota. Our data indicate that genotoxic E. coli may use colibactin to compete for gut niche utilization., The genotoxin colibactin produced by resident bacteria of the gut microbiota may have tumorigenic effect by inducing DNA double-strand breaks in host cells. Yet, the effect of colibactin on gut microbiota composition and functions remains unknown. To address this point, we designed an experiment in which pregnant mice were colonized with the following: (i) a commensal Escherichia coli strain, (ii) a commensal E. coli strain plus a genotoxic E. coli strain, (iii) a commensal E. coli strain plus a nongenotoxic E. coli mutant strain unable to produce mature colibactin. Then, we analyzed the gut microbiota in pups at day 15 and day 35 after birth. At day 15, mice that were colonized at birth with the genotoxic strain showed lower levels of Proteobacteria and taxa belonging to the Proteobacteria, a modest effect on overall microbial diversity, and no effect on gut microbiome. At day 35, mice that received the genotoxic strain showed lower Firmicutes and taxa belonging to the Firmicutes, together with a strong effect on overall microbial diversity and higher microbial functions related to DNA repair. Moreover, the genotoxic strain strongly affected gut microbial diversity evolution of pups receiving the genotoxic strain between day 15 and day 35. Our data show that colibactin, beyond targeting the host, may also exert its genotoxic effect on the gut microbiota. IMPORTANCE Infections of genotoxic Escherichia coli spread concomitantly with urbanized progression. These bacteria may prompt cell senescence and affect DNA stability, inducing cancer via the production of colibactin, a genotoxin shown capable of affecting host DNA in eukaryotic cells. In this study, we show that the action of colibactin may also be directed against other bacteria of the gut microbiota in which genotoxic E. coli bacteria have been introduced. Indeed, the presence of genotoxic E. coli induced a change in both the structure and function of the gut microbiota. Our data indicate that genotoxic E. coli may use colibactin to compete for gut niche utilization.

Made available in DSpace on 2021-06-25T10:45:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-03-01 The aim of this study was to evaluate the prevalence and antimicrobial resistance profile of bacterial species isolated from infected sites of canines. All samples were collected from canine patients who received clinical or surgical care at the veterinary teaching hospital between March 2016 and November 2017. The samples were analyzed in a private pathology laboratory. A descriptive analysis of 295 antimicrobial susceptibility test reports was performed. Staphylococcus spp. (104/295 [35.25%]), Escherichia coli (100/295 [33.90%]), Proteus spp. (44/295 [14.92%]), Pseudomonas spp. (25/295 [8.47%]), and Klebsiella spp. (20/295 [6.78%]) were more frequently isolated, and a high incidence of multidrug resistance was observed (69,83% [206/295]). Methicillin-resistant Staphylococcus spp. accounted for 33% (33/100) of the Staphylococcus strains. Enterobacteriaceae cefotaxime resistance constituted 22.82 ± 4.49% and Enterobacteriaceae imipenem resistance constituted 5% (1/20) for Klebsiella spp., 5% (5/100) for E coli, and 6.82% (3/44) for Proteus spp. Pseudomonas spp. strains accounted for 8% (2/25) of imipenem resistance and 45.45% (10/22) of polymyxin B resistance. Our findings revealed a high rate of multidrug-resistant bacteria involvement in the infectious process of dogs. From the perspective of the One Health scenario, our results showed alarming data, given the high risk of resistant-strain dissemination between animals, owners, and healthcare professionals. There is an urgent need for strategies to control and prevent the evolution of new multidrug-resistant bacteria in veterinary hospitals. It is also crucial to understand and emphasize the role of veterinary professionals in this public health battle. Departament of Veterinary Clinic and Surgery School of Agrarian and Veterinary Sciences São Paulo State University (UNESP/FCAV) Department of Veterinary Pathology School of Agrarian and Veterinary Sciences São Paulo State University (UNESP/FCAV) Departament of Veterinary Clinic and Surgery School of Agrarian and Veterinary Sciences São Paulo State University (UNESP/FCAV) Department of Veterinary Pathology School of Agrarian and Veterinary Sciences São Paulo State University (UNESP/FCAV)

The purpose of the study was to analyze the prevalence of multidrug-resistant (MDR) pathogens (Escherichia coli and Klebsiella pneumonia) isolated from cancer patients. material and methods. 862 strains of E. coli and 1 461 strains of K. pneumoniae isolated from wound, blood, urine and other pathological materials from cancer patients treated at N.N. Blokhin National Medical Research Center of Oncology (Moscow) between 2014 and 2016 were analyzed. results. Over the last 3 years, there has been a significant increase in the number of E.coli resistant to piperacillin-tazobactam (from 8.1 to 25 %), ceftriaxone (from 15.7 to 100 %), ertapenem (from 8.1 to 97.7 %), and amikacin (c 4.5 to 14.6 %). The number of E. coli producing carbapenemases has increased from 5.8 to 20.8 % (p≤0.0001). Among K. pneumoniae, a significant increase in the number of strains resistant to piperacillin-tazobactam (from 59.7 to 89.7 %), ceftazidime (from 62.8 to 82.8 %), ceftriaxone (from 62.8 to 100 %), cefepime (from 63.1 to 83.8 %), ertapenem (from 62.1 to 100 % ), imipenem (from 30.0 to 81.0 %), and meropenem (from 53.9 to 86.8% ) is observed. conclusion. Evaluation of the frequency of isolation of MDR microorganisms over time is an essential element of epidemiological surveillance and is necessary for the correct treatment of patients with infectious complications.

Different molecules have been tested as analog receptors due to their capacity to bind bacteria and prevent cell adhesion. By using in vitro assays, the present study characterized the aqueous and alkaline extracts from microalgae Pavlova lutheri and Pavlova gyrans and evaluated the capacity of these extracts to adhere to enterobacteria (Salmonella Typhimurium). The aqueous and alkaline extracts of both species were fractionated via freeze-thawing, giving rise to soluble and insoluble (precipitate) fractions in cold water. The obtained fractions were studied using thermogravimetric, methylation analyses, and using 1D and 2D NMR techniques. The cold-water-soluble fractions obtained from the aqueous extracts were mainly composed of highly branched (1&rarr, 3),(1&rarr, 6)-&beta, glucans, whereas the cold-water-precipitate fractions were constituted by (1&rarr, 3)-&beta, glucans. The alkaline extract fractions showed similar compositions with a high protein content, and the presence of glycosides (sulfoquinovosylglycerol (SQG), digalactosylglycerol (DGG)), and free fatty acids. The linear (1&rarr, glucans and the alkaline extract fractions showed an adhesion capacity toward Salmonella. The chemical composition of the active fractions suggested that the presence of three-linked &beta, glucose units, as well as microalgal proteins and glycosides, could be important in the adhesion process. Therefore, these microalgal species possess a high potential to serve as a source of anti-adhesive compounds.

Multidrug-resistant bacteria have become common all over the world, necessitating the development of new therapeutic strategies. Synergistic interactions between conventional antibiotics and natural bioactive may have therapeutic benefits in a clinical setting. There are plenty of medicinal plants that have proven efficacy against broad spectrum of micro-organisms. The aim of the work was to assess the antibacterial activity of Cladanthus arabicus, a Moroccan medicinal plant, and Bubonium imbricatum, a Moroccan endemic plant. The evaluation of the synergistic effect of extracted essential oils (EOs) together with some conventional antibiotics was also investigated. Checkerboard test was used to evaluate the interaction of EOs in combination with amoxicillin and neomycin. The results showed that EOs contain a potent activity against the tested Enterobacteriaceae isolates, with inhibition zones values in the range of 8·05 ± 0·1 and 13·1 ± 0·11 mm and MIC values between 200 μg ml-1 to 800 μg ml-1 for C. arabicus and from 400 μg ml-1 to 1600 μg ml-1 for B. imbricatum, respectively. Moreover, the current study allowed concluding that both EOs showed not only satisfactory antibacterial properties but also active effects combined with conventional antibiotics demonstrated by the Fractional Inhibitory Concentration Index (FICI). These findings are very interesting since there are no previous studies on synergistic interactions of these two plants with antibiotics. Significance and impact of the study The development of antibiotic resistance is multifactorial, including the specific nature of the relationship of bacteria to antibiotics. This situation has forced scientists to search for new antimicrobial substances from various sources as novel antimicrobial chemotherapeutic agents. Recently, medicinal plants and their derivatives (essential oils, extracts) have become very important in therapeutics because they encounter minimal challenges of the emergence of resistance. In this direction, the antimicrobial activity of the endemic Bubonium imbricatum plant and medicinal Cladanthus arabicus plant essential oils against multidrug-resistant Enterobacteriaceae strains was demonstrated.

Background Citrobacter strains are opportunistic pathogens often responsible for serious enteric as well as extra-intestinal diseases, and therefore the O-antigenic scheme, still in use in diagnostic identification, should be set for proper serotyping. The structures of more than 30 different Citrobacter O-antigens (O-polysaccharide chains of the lipopolysaccharides) of 43 Citrobacter O-serogroups have been elucidated so far. However, relationships between strains in several heterogeneous serogroups still need to be clarified by immunochemical studies. These include complex serogroups O3 and O8, represented by 20 and 7 strains, respectively, which are the subject of the present work. Earlier, the O-polysaccharide structures have been determined for Citrobacter O3 strain Be35/57 (PCM 1508) and Citrobacter O8 strain Be64/57 (PCM 1536). Results Serological studies (immunoblotting) carried out on Citrobacter lipopolysaccharides from different strains ascribed to serogroups O3 and O8 showed that each of these serogroups should be divided into non-cross-reacting subgroups. Based on the results of chemical analyses and 1H and 13C NMR spectroscopy the structure of Citrobacter O-antigens from strains PCM 1504 (O6) and PCM 1573 (O2) have been established. Chemical data combined with serological analyses showed that several Citrobacter strains should be reclassified into other serogroups. Conclusions Immunochemical studies carried out on Citrobacter LPS, described in this paper, showed the expediency of reclassification of: 1) strains PCM 1504 and PCM 1573 from serogroups O6 and O2 to serogroups O3 and O8, respectively, 2) strains PCM 1503 and PCM 1505 from serogroups O3 and O8 to new serogroups O3a and O8a, respectively.

Studies evaluating the occurrence of enteropathogenic bacteria in urban rats (Rattus spp.) are scarce worldwide, specifically in the urban environments of tropical countries. This study aims to estimate the prevalence of diarrhoeagenic Escherichia coli (DEC) and Salmonella spp. with zoonotic potential in urban slum environments. We trapped rats between April and June 2018 in Salvador, Brazil. We collected rectal swabs from Rattus spp., and cultured for E. coli and Salmonella spp., and screened E. coli isolates by polymerase chain reaction to identify pathotypes. E. coli were found in 70% of Rattus norvegicus and were found in four Rattus rattus. DEC were isolated in 31.3% of the 67 brown rats (R. norvegicus). The pathotypes detected more frequently were shiga toxin E. coli in 11.9%, followed by atypical enteropathogenic E. coli in 10.4% and enteroinvasive E. coli in 4.5%. From the five black rats (R. rattus), two presented DEC. Salmonella enterica was found in only one (1.4%) of 67 R. norvegicus. Our findings indicate that both R. norvegicus and R. rattus are host of DEC and, at lower prevalence, S. enterica, highlighting the importance of rodents as potential sources of pathogenic agents for humans.

Introduction:the emergence of extended-spectrum beta-lactamase-producing Enterobacteriaceae (E-ESBLs) is currently a major public health problem in the world and, in particular, in developing countries. In Cameroon, data on E-ESBLs are rare, especially in Garoua and in the northern region of the country. The objective of this study is to document the epidemiology of E-ESBL infections in small children and to explore their associations with possible risk factors. Methods:this was a cross-sectional, descriptive study conducted from June 14 to September 30, 2018, including small children with suspected urinary tract infections (UTI) attending the outpatient pediatric departments of two health facilities in the city of Garoua. Urine samples were analyzed at the Bacteriology Laboratory of the Pasteur Center of Cameroon, Annex Garoua. Bacterial culture was carried out on Bio-Rad UriSelect® chromogenic agar and the identification was confirmed by bioMérieux API 20E. The antibiotic susceptibility was determined using the bioMérieux ATB UR gallery and the ESBL phenotype was detected by the double disk synergy method according to the CA-SFM 2013 recommendations. The data was analyzed with the R Statistical Software version 2.15.2. Results:a total of 57 urine samples were collected from children aged from one month to two years, 37 boys and 20 girls. Bacteria were detected by culture in 20 samples:Escherichia coliwas the most frequently (75 %) isolated species followed byKlebsiella pneumoniae(25%). More than half of the infected samples (55%) contained E-ESBL. The presence of an ESBL was significantly associated with previous antibiotic intake up to 3 months prior current UTI (p=0.01664). The E-ESBL strains showed co-resistance to different antibiotics. Conclusion:this study reveals the important dissemination of E-ESBLs among small children in the community and a high rate of co-resistance to the different antibiotic families commonly used.

Salmonella enterica is one of the most common pathogens associated with produce outbreaks worldwide; nonetheless, the mechanisms uncovering their interaction with plants are elusive. Previous reports demonstrate that S. enterica ser. Typhimurium (STm), similar to the phytopathogen Pseudomonas syringae pv. tomato (Pst) DC3000, triggers a transient stomatal closure suggesting its ability to overcome this plant defense and colonize the leaf apoplast. In order to discover new molecular players that function in the stomatal reopening by STm and Pst DC3000, we performed an Arabidopsis mutant screening using thermal imaging. Further stomatal bioassay confirmed that the mutant plants exo70h4-3, sce1-3, bbe8, stp1, and lsu2 have smaller stomatal aperture widths than the wild type Col-0 in response to STm 14028s. The mutants bbe8, stp1 and lsu2 have impaired stomatal movement in response to Pst DC3000. These findings indicate that EXO70H4 and SCE1 are involved in bacterial-specific responses, while BBE8, STP1, and LSU2 may be required for stomatal response to a broad range of bacteria. The identification of new molecular components of the guard cell movement induced by bacteria will enable a better understanding of the initial stages of plant colonization and facilitate targeted prevention of leaf contamination with harmful pathogens.

International audience; Acquired carbapenemases currently pose one of the most worrying public health threats related to antimicrobial resistance. A NDM-1-producing Salmonella Corvallis was reported in 2013 in a wild raptor. Further research was needed to understand the role of wild birds in the transmission of bacteria resistant to carbapenems. Our aim was to investigate the presence of carbapenem-resistant Escherichia coli in gulls from southern France. In 2012, we collected 158 cloacal swabs samples from two gull species: yellow-legged gulls (Larus michahellis) that live in close contact with humans and slender-billed gulls (Chroicocephalus genei) that feed at sea. We molecularly compared the carbapenem-resistant bacteria we isolated through culture on selective media with the carbapenem-susceptible strains sampled from both gull species and from stool samples of humans hospitalized in the study area. The genes coding for carbapenemases were tested by multiplex PCR. We isolated 22 carbapenem-resistant E. coli strains from yellow-legged gulls while none were isolated from slender-billed gulls. All carbapenem-resistant isolates were positive for blaVIM-1 gene. VIM-1-producing E. coli were closely related to carbapenem-susceptible strains isolated from the two gull species but also to human strains. Our results are alarming enough to make it urgently necessary to determine the contamination source of the bacteria we identified. More generally, our work highlights the need to develop more bridges between studies focusing on wildlife and humans in order to improve our knowledge of resistant bacteria transmission routes.

Microorganisms, including the bacterial populations living inside plant tissues (endophytes), and vegetation are components of constructed wetland (CW) systems playing pivotal roles in the water treatment process and in the ecosystem establishment. The present study focuses on the assessment of the culturable endophytic bacterial communities of Canna flaccida plants, the dominant plant species in a polycultured CW treating wastewater from a tourism facility. Bacterial endophytes were grouped by random amplified polymorphic DNA and identified by 16S ribosomal RNA gene sequencing. From the bacterial isolates, 103 were considered for phylogenetic analysis, falling in 25 genera within the gamma-, beta- alpha-Proteobacteria, Flavobacteria, Sphingobacteria, Actinobacteria and Bacilli classes. Forty-nine percent of the isolates belonged to the Enterobacteriaceae family, suggesting that the plants in CW systems may act as a sink of potential human pathogenic microorganisms; nevertheless their abundance on the plant tissues was reduced from the inlet (62%) to the outlet zone (38%). High diversity in terms of genera was found in C. flaccida tissues, although the number of genera common to both sampling zones was low, which suggests that the processes occurring within the CW, including the water characteristics gradient from the inlet to the outlet (e.g. total suspended solids, organics and fecal loads), had the ability to shape the diversity of the endophytic communities. (C) 2016 Elsevier All rights reserved. C.S.C. Calheiros and S.I.A. Pereira thank the grants SFRH/BPD/63204/2009 and SFRH/BPD/65134/2009, respectively, from Fundação para a Ciência e Tecnologia (FCT, Portugal), Fundo Social Europeu and Fundos Nacionais do MEC through the program QREN−POPH−Tipologia 4.1-Formação Avançada. This work was supported by National Funds from FCT−Fundação para a Ciência e a Tecnologia through project UID/Multi/50016/2013. The authors are thankful for the collaboration of Paço de Calheiros and Saint-Gobain Weber Portugal, S.A. The authors wish to thank Dr. Pedro Carvalho for reviewing the manuscript.

We propose the main characteristics of a new bacterium species named Nissabacter archeti strain 2134 (CSURP3445 = LT631518), isolated from pustule scalp of a 29-year-old man at hospital Archet 2, Nice, south of France.

The bacterial communities that are established during natural meat fermentation depend on the processing conditions and the type of meat substrate used. Six pork samples of variable quality (reflected in pH values) and six less conventional meats (beef, horse, hare, wild deer, wild duck, and wild boar) were naturally fermented under controlled conditions in model systems. The development of lactic acid bacteria (LAB), coagulase-negative staphylococci (CNS), and enterobacteria was followed using culture-dependent techniques and (GTG)5-PCR fingerprinting of genomic DNA from the isolates obtained. Taken together, Latilactobacillus sakei was the most abundant LAB species, although Latilactobacillus curvatus was more manifest in high-pH pork. Within staphylococci, common species were encountered (i.e., Staphylococcus equorum, Staphylococcus saprophyticus, and Staphylococcus xylosus), although some atypical ones (i.e., Staphylococcus succinus) were also recovered. Within enterobacteria, Serratia spp. prevailed in more acidic pork batches and in beef, whereas Hafnia spp. prevailed in game meat fermentations. Enterobacterial counts were particularly high in fermentations with low acidity, namely for some pork batches, hare, wild duck, and wild boar. These findings should be considered when naturally fermented meat products are manufactured, as the use of game meat or meat with high pH can give rise to safety concerns.

In order to investigate enterobacteria presence involved in the secondary infections in Porcine Reproductive and Respiratory Syndrome (PRRS) pigs with different viral co-infections, we identified enterobacteria for guiding clinical treatment. Twenty-one diseased pigs were diagnosed with the PRRS virus (PRRSV) and other 7 virus primers by PCR/RT-PCR in the lung and spleen samples. Enterobacteria were isolated using MacConkey agar from 5 visceral samples of PRRS pigs, and identified by 16S rDNA sequencing. PRRSV was positive in 100% of the lung samples and 81.0% of the spleen samples. Seven diseased pigs were diagnosed with only PRRSV infection (33.3%), 7 pigs with PRRSV and 1 or 2 other viruses (33.3%) and 7 pigs with PRRSV and more than 2 types of other viruses (33.3%). PRRSV was more inclined to co-infect pigs with porcine group A rotavirus (PARV) with the co-infection rate of 52.4% (11/21). Approximately 13 types of bacteria were successfully isolated from lung, spleen, liver, kidney and lymph node samples of different PRRS pigs. Enterobacteria were isolated in 100% of lung, liver and lymph samples from pigs infected with PRRSV alone. However, the isolation rates were significantly decreased in the more than 3 viruses co-infection group. Escherichia coli was the most prevalent bacterium, followed by Morganella, Proteus, Shigella, Salmonella, Klebsiella and Aeromonas. Most of the isolated enterobacteria were opportunistic pathogens. Therefore, timely combination with antimicrobial agents is necessary for effective treatment of PRRS-infected pigs., Highlights • PRRSV was more inclined to co-infect with PARV. • Pigs co-infected with more virus, less enterobacteria were isolated from their viscera. • Enterobacteria isolated from viscera of PRRS pigs were opportunistic pathogens. • coli, Morganella and Proteus were the most prevalent enterobacteria. • Secondary infection of enterobacteria should be considered in PRRS treatment.

Domestic environment, in particular, kitchen setting is a well-established source of microbial contamination. Kitchen sponges represent an important vehicle of microbial transmission and maintenance of spoilage bacteria and pathogenic strains responsible for food borne diseases. The aim of this study was to evaluate the microbial communities of 100 ‘in-use’ kitchen sponges, improving the knowledge on their role in cross-contamination in domestic environment and transmission of ESBLproducing strains. Sponges were processed for: aerobic mesophilic bacteria (AMB), Enterobacteriaceae (EB), yeasts and molds (YM), coagulase-positive staphylococci (CPS), micrococci (MCC), anaerobic sulfite reducing bacteria (ASR), and for the detection of Listeria monocytogenes, Salmonella spp. and Yersinia enterocolitica. A total of 309 enterobacteria strains were identified and then processed for ESBL (Extended Spectrum Beta Lactamase) phenotypical expression. A high contamination level of kitchen sponges was observed (mean value AMB 8.25±1.1; EB 5.89±1.2; YM 5.57±1.1; MCC 4.82±0.1 log CFU/g). Identified enterobacteria strains revealed several opportunistic and pathogenic agents such as Enterobacter cloacae (28%), Citrobacter freundii (23.3%), Cronobacter sakazakii (14.6%) and other strains in lower percentage. Listeria monocytogenes was found in only one sponge (1%). A total of 69 (22.3%) enterobacteria resulted ESBL+, with the following prevalence: P. rettgeri (50%), L. adenocarboxilata (30%), K. pneumoniae (25%), K. oxytoca (25%), C. sakazakii (20%), E. cloacae (20.7%), C. freundii (20.1%). Results confirm the potential role of kitchen sponges as vehicle for food-borne pathogens such as, C. sakazakii for the first time, infectious agents and spoilage microorganisms. The observed high contamination level and the presence of several ESBLs opportunistic pathogens, stresses the necessity to improve a proper education of the consumers on the effective treatment to reduce their microbial loads.

Acidic beers, such as Belgian lambic beers and American and other coolship ales, are becoming increasingly popular worldwide thanks to their refreshing acidity and fruity notes. The traditional fermentation used to produce them does not apply pure yeast cultures but relies on spontaneous, environmental inoculation. The fermentation and maturation process is carried out in wooden barrels and can take up to three years. It is characterized by different microbial species belonging to the enterobacteria, acetic acid bacteria, lactic acid bacteria, and yeasts. This review provides an introduction to the technology and four fermentation strategies of beer production, followed by the microbiology of acidic beer production, focusing on the main microorganisms present during the long process used for the production of Belgian lambic beers. © 2018 Society of Chemical Industry.

Background: Despite a strong association between Salmonella isolation and slaughter hygiene, as measured by the Enterobacteriaceae levels on pre-chill carcass surfaces, a high variation in this association was observed between sampling days within the same slaughterhouse. It was hypothesised that in a scenario of high exposure on the farm, batches with a high prevalence of carrier pigs shedding a high number of Salmonella may enhance the risk of contamination on some slaughter days. Thus, the aim of this study was to assess the profile of Salmonella carried in the intestinal contents of slaughter pigs. Materials, Methods & Results: Ten pig batches slaughtered in a slaughterhouse were investigated for the presence of Salmonella. From each pig, the following samples were taken: i. blood collected at bleeding; ii. sponges rubbed on the carcass surface after bleeding and before chilling; iii. fragment of the ileocecal region of the intestine. Serum samples were subjected to a ELISA-Typhimurium test. Sponges were investigated for the presence of Salmonella and total aerobic mesophilic (TAM) and terobacteriaceae (EC) bacterial counts. Salmonella was enumerated in the intestinal contents. Selected Salmonella strains were subjected to an antimicrobial resistance disk diffusion test, macro-restriction with Xba-I (PFGE) and whole genome sequencing (WGS). From the 50 sampled pigs, 96% were positive in the ELISA-Typhimurium test and 64% were Salmonella-positive in the intestinal contents. The amount of Salmonella in the intestinal content samples was highly variable, and the mean log of fitted distributions of Salmonella in the batch ranged from -2.97 to 2.25 cfu.g-1. The slaughter process achieved a logarithmic reduction, ranging from 0.64 to 2.35 log cfu.cm-2 for TAM and from 0.55 to 2.57 log cfu.cm-2 for EC. Salmonella was isolated from 16% of the carcasses after bleeding; this frequency decreased to 8% at the pre-chill step. All positive pre-chill carcasses originated from pigs carrying Salmonella in the intestinal content and from batches with a high number of carrier pigs. Salmonella Typhimurium and its monophasic variant were the most frequent in the intestinal contents and carcasses. Resistance was detected against ampicillin (42.5%), tetracycline (42.5%), sulfonamide (40%), gentamicin (25%) and ciprofloxacin (12.5%). Regarding colistin, 85% of the tested strains were classified as non-susceptible. The monophasic variant S. Typhimurium strains subjected to PFGE and WGS presented different profiles; several antimicrobial resistance genes were identified and all belonged to ST-19. Discussion: In this study, almost all sampled pigs entering the slaughter line had been exposed to Salmonella on the farm and a high number were carrying Salmonella in their guts. While the three batches with Salmonella-positive carcasses at the pre-chill step presented TAM media that was not significantly different from the other batches, there was a higher number of positive pigs carrying Salmonella in their intestinal contents. Moreover, the batch with the highest number of positive carcasses also presented the highest Salmonella mean count in their intestinal contents. The profile of Salmonella carried in the intestinal content of slaughter pigs proved to be highly variable in terms of the frequency, number of bacteria, serovars, antimicrobial resistance, and genotypes. Results indicate that the day-to-day variability in the prevalence and number of Salmonella in the intestinal contents of slaughter batches is likely to influence the frequency of contaminated pre-chill carcasses. Salmonella Typhimurium isolated from the intestinal contents of slaughter pigs may belong to genotypes involved in human disease and may carry several antimicrobial resistance genes. These aspects should be taken into account when planning Salmonella control in swine. Made available in DSpace on 2019-10-31T18:14:00Z (GMT). No. of bitstreams: 1 final9102.pdf: 262646 bytes, checksum: 0c65477537e69c9e49d13aafe1556515 (MD5) Previous issue date: 2019

The Enterobacterales order contains 7 families with more than 40 bacterial genera which are isolated from various types of environment. The genera Shimwellia, Obesumbacterium, Rahnella, Citrobacter, Klebsiella, Raoultella, Serratia and Enterobacter are often found in the beer brewing process. The ability of these bacteria to grow and multiply in the finished beer is minimal. The bacteria occur primarily as contamination of yeast. They are harmful because these bacteria produce sensory undesirable substances (e.g. dimethylsulfide, diacetyl, acetoin) at the beginning of the main fermentation and get into the finished beer. Some species are involved in the formation of N-nitrosamines harmful for health. Here we present an overview of the basic morphological and physiological properties of these bacteria and describe their importance from the point of view of harmfulness in the beer brewing process.

RESUMEN La resistencia a las polimixinas mediada por plásmidos (gen mcr-1) representa una amenaza para la salud pública, puesto que colistina es utilizada en la práctica médica como una de las últimas alternativas para el tratamiento de gérmenes multiresistentes. Este estudio describe la circulaciónde cepas de Enterobacterias que portan este gen de resistencia, aisladas de pacientes hospitalizados, así como también de la comunidad. Los hallazgos de la Red de Vigilancia de la Resistencia a los Antimicrobianos-Paraguay fueron de casi el 5 % (4,7) en cepas remitidas con criterio de sospecha, siendo las especies involucradas Escherichiacoli, Klebsiella pneumoniae y Salmonella Schwarzengrund. Además, por métodos moleculares se confirmaron en todas ellas la portación de otros genes de resistencia (KPC, CTX-M, Qnr B, Qnr S, aac (6`)-Ib-cr) asociados al mcr-1. ABSTRACT Resistance to polymyxins mediated by plasmids (mcr-1 gene) represents a threat to public health, since colistin is used in medical practice, as one of the last alternatives, for the treatment of multi-resistant germs. This study describes the circulation of strains of Enterobacteria that carry this resistance gene, isolated from hospitalized patients, as well as from the community. The findings of the Red de Vigilancia de la Resistencia a los Antimicrobianos-Paraguay were almost 5% (4.7) in strains submitted with suspicion criteria; the species involved being Escherichia coli, Klebsiella pneumoniae and Salmonella Schwarzengrund. In addition, molecular methods confirmed in all of them the carrying of other resistance genes (KPC, CTX-M, Qnr B, Qnr S, aac (6`)-Ib-cr) associated with mcr-1.

Made available in DSpace on 2018-12-11T17:24:20Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-04-01. Added 1 bitstream(s) on 2019-10-09T18:32:06Z : No. of bitstreams: 1 S1516-635X2018000200189.pdf: 150511 bytes, checksum: b0bdba2ae71fc325d8ad9a2341d5bca4 (MD5) Raoultella ornithinolytica is a gram-negative aerobic bacterium belonging to Enterobacteriaceae family, an emerging pathogen that causes several pathogenic conditions in man, with little veterinary importance; however, its identification is underestimated by conventional laboratory techniques. The present study reports the identification of R. ornithinolytica in Tinamus solitaries, during a routine sanitary evaluation of aerobic enterobacteria in cloacal microbiota of birds belonging to the Güira Oga Center, Puerto Iguazu, Argentina. The sample was preliminary classified as Klebsiella spp.; however, after the use of the MALDI-TOF MS technique it was identified as R. ornithinolytica. The sample was submitted to an antimicrobial susceptibility test, where it showed a similar pattern profile as reported in the literature, with resistance to ampicillin and other β-lactam antibiotics. It is possible that Raoultella spp are more common in birds as it is reported. Therefore, review studies on bacteria collections of avian origin, as well as cases with confirmation of Klebsiella, should be deeply evaluated in laboratorial routine, mainly due to the pathogenic potential of R. ornithinolytica for Poultry, as well as for public health. Faculdades Integradas de Ourinhos - FIO, R. Luzia de Masseno Pontes 430, Bairro Jardim Itamaraty Ourinhos Centro Güira Oga Universidade Estadual Paulista - UNESP Instituto Federal de Ciências e Tecnologia do Amazonas - IFAM Universidade de São Paulo - USP Universidade Estadual Paulista - UNESP

A new, less restrictive definition increases detection of Klebsiella pneumoniae carbapenemase producers., Preventing transmission of carbapenemase-producing, carbapenem-resistant Enterobacteriaceae (CP-CRE) is a public health priority. A phenotype-based definition that reliably identifies CP-CRE while minimizing misclassification of non–CP-CRE could help prevention efforts. To assess possible definitions, we evaluated enterobacterial isolates that had been tested and deemed nonsusceptible to >1 carbapenem at US Emerging Infections Program sites. We determined the number of non-CP isolates that met (false positives) and CP isolates that did not meet (false negatives) the Centers for Disease Control and Prevention CRE definition in use during our study: 30% (94/312) of CRE had carbapenemase genes, and 21% (14/67) of Klebsiella pneumoniae carbapenemase–producing Klebsiella isolates had been misclassified as non-CP. A new definition requiring resistance to 1 carbapenem rarely missed CP strains, but 55% of results were false positive; adding the modified Hodge test to the definition decreased false positives to 12%. This definition should be considered for use in carbapenemase-producing CRE surveillance and prevention.

When introducing a novelty food, its safety needs to be monitored. One of the safety aspects of human health is microbial contamination. In this work, microbiological parameters of long-term stored edible insect material - mealworm (Tenebrio molitor), lesser mealworm (Alphitobius diaperinus), field cricket (Gryllus assimilis), and migratory locust (Locusta migratoria) were evaluated. The monitored indicators (colony forming units, enterobacteria, lactic acid bacteria, yeasts and moulds) were evaluated using common microbiological methods. All samples of stored insect were determined as safe for human consumption, except for the lesser mealworm sample from 2016, in which case the limit was exceeded. Sample of adult field cricket seems to be suitable for long-term storage, as it contained the lowest amount of microorganism. Sample of dried Gryllus assimilis from 2014 had the lowest microbial contamination. Further results suggest that, for long-term storage, the most suitable way of preparation is killing with boiling water, drying at 103 °C for 12 hours and subsequent hermetic packaging. Při zavádění nové potraviny je potřeba sledovat její bezpečnost. Jedním z bezpečnostních aspektů lidského zdraví je mikrobiální kontaminace. V této práci byly hodnoceny mikrobiologické parametry dlouhodobě skladovaného jedlého materiálu hmyzu - Tenebrio molitor, menšího jedle (Alphitobius diaperinus), polní kriket (Gryllus assimilis) a migrující kobylka (Locusta migratoria). Sledované ukazatele (jednotky tvořící kolonie, enterobakterie, bakterie mléčného kvašení, kvasinky a plísně) byly hodnoceny za použití běžných mikrobiologických metod. Všechny vzorky uloženého hmyzu byly stanoveny jako bezpečné pro lidskou spotřebu, s výjimkou menšího vzorku červů z roku 2016, kdy byl limit překročen. Vzorek kriketu pro dospělé pole se zdá být vhodný pro dlouhodobé uchovávání, protože obsahoval nejnižší množství mikroorganismů. Vzorek sušeného Gryllus assimilis od roku 2014 měl nejnižší mikrobiální kontaminaci. Další výsledky naznačují, že pro dlouhodobé skladování je nejvhodnější způsob přípravy zabíjení vroucí vodou, sušení při 103 ° C po dobu 12 hodin a následné hermetické balení. (automatický překlad)

Background:Edwardsiella tarda infections are frequent causes of severe outbreaks in the fish farming industry besides representing possible zoonotic risks. However, naturally occurring outbreaks that affect various species besides fishes are seldom described. Aim: To report an outbreak of acute mortality caused by E. tarda affecting multiple species that inhabited a natural pond in the state of São Paulo, Brazil. Materials and methods: Three adult tilapias, three Mallard ducks and one Snow egret were necropsied and subjected to further microbiological tests. Gross and microscopic lesions were documented. The antibiotic susceptibility and phylogenetic similarities among fish and avian strains were also determined. The E. tarda species was confirmed through MALDI-TOF, partial sodB sequencing and phylogenetic analysis. Results: Macroscopical findings between the three species included intestinal dilatation, mucosal hyperaemia and mucous to liquid contents. Common histopathology findings included acute enteritis, increased number of intraepithelial lymphocytes with bacteria adhered to the intestinal epithelium and lymphoid depletion in the spleen. E. tarda was isolated from several organs from all affected species. The phylogeny employing amplified fragment length polymorphism (AFLP) of eleven strains revealed high similarity (>90%) among the isolates regardless of the affected species or sampled organs. Ten isolates of E. tarda showed susceptibility to all tested antibiotics. Conclusions:E. tarda was identified as the cause of death of the species examined. Further studies would be necessary to determine the virulence of these strains and the possible risks regarding public health.

The class of ß-lactam antibiotics has proven highly efficient in targeting bacterial penicillin-binding proteins (PBP) leading to the blocking of the bacterial cell wall synthesis. However, the benefit of these drugs is limited because of bacterial resistance mechanisms; the most widespread resistance involves ß-lactamase enzymes (ßLACT) that inactivate ß-lactam-based molecules. We focused on PBPs and ßLACTs from enterobacteria, and performed a detailed in silico study of PBPs whose inactivation is lethal for the bacteria and of ßLACTs that have a PBP-type catalytic mechanism. The comparison of the sequences and structures of PBPs and ßLACTs shows an almost perfect conservation of the catalytic site, and a high spatial resemblance of the whole functional cavity despite a very low overall sequence identity. Some notable differences in the functional cavity were observed in the vicinity of the catalytic site: four tyrosines are well conserved in the PBPs, whereas the residues occurring at equivalent positions in the ßLACT families present other physicochemical properties. These tyrosines are thus good candidates to be targeted in designing new antibiotic molecules with increased affinity and specificity for PBPs, with the goal of overcoming drug resistance. Our analysis also identified residues that have similar characteristics in most ßLACT families and different properties in PBPs; these are interesting targets for new ligands that specifically inhibit ßLACT proteins. The in silico approach presented here can be extended to other protein systems in view of guiding and improving rational drug design., SCOPUS: ar.j, info:eu-repo/semantics/published

Background Citrobacter strains are opportunistic pathogens often responsible for serious enteric as well as extra-intestinal diseases, and therefore the O-antigenic scheme, still in use in diagnostic identification, should be set for proper serotyping. The structures of more than 30 different Citrobacter O-antigens (O-polysaccharide chains of the lipopolysaccharides) of 43 Citrobacter O-serogroups have been elucidated so far. However, relationships between strains in several heterogeneous serogroups still need to be clarified by immunochemical studies. These include complex serogroups O3 and O8, represented by 20 and 7 strains, respectively, which are the subject of the present work. Earlier, the O-polysaccharide structures have been determined for Citrobacter O3 strain Be35/57 (PCM 1508) and Citrobacter O8 strain Be64/57 (PCM 1536). Results Serological studies (immunoblotting) carried out on Citrobacter lipopolysaccharides from different strains ascribed to serogroups O3 and O8 showed that each of these serogroups should be divided into non-cross-reacting subgroups. Based on the results of chemical analyses and 1H and 13C NMR spectroscopy the structure of Citrobacter O-antigens from strains PCM 1504 (O6) and PCM 1573 (O2) have been established. Chemical data combined with serological analyses showed that several Citrobacter strains should be reclassified into other serogroups. Conclusions Immunochemical studies carried out on Citrobacter LPS, described in this paper, showed the expediency of reclassification of: 1) strains PCM 1504 and PCM 1573 from serogroups O6 and O2 to serogroups O3 and O8, respectively, 2) strains PCM 1503 and PCM 1505 from serogroups O3 and O8 to new serogroups O3a and O8a, respectively.

There is growing evidence indicating that drug-resistant bacteria notably Extended spectrum β-lactamases producing Enterobacteria are greatly implicated in immunocompromised groups namely HIV-infected patients. Little is known about the burden of ESPL-E in places where HIV infection is rampant. Therefore, the study was aimed to assess the prevalence, antimicrobial resistance pattern of ESPL-E among HIV/AIDS patients. A cross-sectional study was conducted among HIV/AIDS patients seeking Antiretroviral Treatment (ART) service at the University of Gondar Hospital from February–May, 2017. The pretested and -structured questionnaire was used to collect data on sociodemographic and clinicalrelated factors. Clean catch midstream urine samples were collected and cultured in line with standard procedures. The drug susceptibility testing was performed by Kirby Bauer disc diffusion method. Extended-spectrum β-lactamase (ESBL) detection was performed using a doubledisc synergy test and combined disc methods. Data entry and analysis were performed using SPSS version 20. Among a total of 387 HIV/AIDS patients, 42 (10.9%) Enterobacteria uropathogens were identified. Among these isolates, nine (21.4%) were ESBL producers. The highest prevalence of ESBL production was Escherichia coli (44.4%) followed by Klebsiella pneumoniae (22.2%) and Enterobacter spp. (22.2%). Higher drug resistance rates were observed among ESBL-producing isolates compared to ESBL-nonproducing isolates. Amox-clavulanic (100%), ampicillin (95%), cotrimoxazole (74%), cefotaxime (88.9%), and ceftazidime (88.9%) had high resistance rates to Extended spectrum β-lactamases producing Enterobacteria. The overall prevalence of multidrug resistance of all isolates was 92.9%, and all the ESBL isolates were multidrug resistant. Therefore, antimicrobial stewardship programs needed to be promoted for the rational use of drugs especially in the management of HIV/AIDS patients.

Background Breast milk can turn pink with Serratia marcescens colonization, this bacterium has been associated with several diseases and even death. It is seen most commonly in the intensive care settings. Discoloration of the breast milk can lead to premature termination of nursing. We describe two cases of pink-colored breast milk in which S. marsescens was isolated from both the expressed breast milk. Antimicrobial treatment was administered to the mothers. Return to breastfeeding was successful in both the cases. Conclusions Pink breast milk is caused by S. marsescens colonization. In such cases,early recognition and treatment before the development of infection is recommended to return to breastfeeding.

Background: Autotransporter adhesins reach the bacterial cell surface by a complex mechanism. Results: In the case of the autotransporter SadA from Salmonella, a lipoprotein assists in surface display. Conclusion: The similarity to eukaryotic MATH domains suggests that the lipoprotein assists in trimerization of SadA. Significance: Understanding the similarities between autotransport systems might lead to new ways of inhibiting bacterial adhesion., Trimeric autotransporter adhesins (TAAs) are important virulence factors of many Gram-negative bacterial pathogens. TAAs form fibrous, adhesive structures on the bacterial cell surface. Their N-terminal extracellular domains are exported through a C-terminal membrane pore; the insertion of the pore domain into the bacterial outer membrane follows the rules of β-barrel transmembrane protein biogenesis and is dependent on the essential Bam complex. We have recently described the full fiber structure of SadA, a TAA of unknown function in Salmonella and other enterobacteria. In this work, we describe the structure and function of SadB, a small inner membrane lipoprotein. The sadB gene is located in an operon with sadA; orthologous operons are only found in enterobacteria, whereas other TAAs are not typically associated with lipoproteins. Strikingly, SadB is also a trimer, and its co-expression with SadA has a direct influence on SadA structural integrity. This is the first report of a specific export factor of a TAA, suggesting that at least in some cases TAA autotransport is assisted by additional periplasmic proteins.

Introduction. Beta-lactam antibiotics are the most commonly used antibacterial drugs. However, many bacteria have developed resistance to these antibiotics, and the most common form of resistance is the production of beta-lactamase enzymes. Many members of the Enterobacteriaceae family produce different types of these enzymes. Objective. The aim of this study was to perform phenotypic detection of production and identification of beta-lactamase type in Enterobacteriaceae isolated from different clinical specimens from patients hospitalized in the Clinical Center of Serbia. Methods. The strains of Enterobacteriaceae were collected between November 2011 and January 2012 in the laboratory of the Clinical Center of Serbia. The isolates were identified according to the standard microbiology procedures and confirmed by the Vitek2 automated system. Antimicrobial susceptibility testing was performed by the disk diffusion method, and the phenotypic detection of production and identification of betalactamases was performed according to previously described methodologies. Results. In this study, a total of 172 Enterobacteriaceae strains were isolated. Further testing was performed on 54/145 (37.2%) strains showing decreased susceptibility to beta-lactam antibiotics: 13/85 (15.3%) Escherichia coli, 31/46 (67.4%) Klebsiella pneumoniae and 10/14 (71.4%) Proteus mirabilis. Among them, 40/145 (27.6%) strains produced extended spectrum betalactamases (ESBLs), 9/145 (6.2%) - AmpC, 1/145 (0.7%) - K1 beta-lactamase and 4/145 (2.8%) - carbapenemases. Carbapenemases were predominantly detected in K. pneumoniae (75%). Conclusion. Enterobacteriaceae produce different types of betalactamases, and the most common type in our study was ESBLs. Production of carbapenemases detected in Enterobacteriaceae is also an associated problem. [Projekat Ministarstva nauke Republike Srbije, br. ON 175039]

International audience; Reports of carbapenemase-producing Enterobacteriaceae in Africa remain rare and assess mostly blaOXA-48-producing isolates from Mediterranean countries and South Africa. We identified blaNDM-7-producing Enterobacteriaceae in Gabon in 2016. The isolates contained blaNDM-7 IncX3 plasmids that were unusual and similar to the one described in a colistin-resistant Klebsiella pneumoniae SZ04 isolate from China.

Strains of Escherichia coli bearing the pks genomic island synthesize the genotoxin colibactin. Exposure of eukaryotic cells to E. coli producing colibactin induces DNA damages, ultimately leading to cell cycle arrest, senescence and death. Here we describe a simple method to demonstrate the genotoxicity of bacteria producing colibactin following a short infection of cultured mammalian cells with pks(+) E. coli.

International audience; Complete sequences of [i]qnr[/i]D-carrying plasmids in [i]Proteus[/i] spp. collected from French animals

In the context of requested decrease of antimicrobial use in veterinary medicine, our objective was to assess the impact of two doses of marbofloxacin administered on young bulls (YBs) and veal calves (VCs) treated for bovine respiratory disease, on the total population of Enterobacteriaceae in gut flora and on the emergence of resistant Enterobacteriaceae. In two independent experiments, 48 YBs from 6 commercial farms and 33 VCs previously colostrum deprived and exposed to cefquinome were randomly assigned to one of the three groups LOW, HIGH, and Control. In LOW and HIGH groups, animals received a single injection of, respectively, 2 and 10 mg/kg marbofloxacin. Feces were sampled before treatment, and at several times after treatment. Total and resistant Enterobacteriaceae enumerating were performed by plating dilutions of fecal samples on MacConkey agar plates that were supplemented or not with quinolone. In YBs, marbofloxacin treatment was associated with a transient decrease in total Enterobacteriaceae count between day (D)1 and D3 after treatment. Total Enterobacteriaceae count returned to baseline between D5 and D7 in all groups. None of the 48 YBs harbored marbofloxacin-resistant Enterobacteriaceae before treatment. After treatment, 1 out of 20 YBs from the Control group and 1 out of 14 YBs from the HIGH group exhibited marbofloxacin-resistant Enterobacteriaceae. In VCs, the rate of fluoroquinolone-resistant Enterobacteriaceae significantly increased after low and high doses of marbofloxacin treatment. However, the effect was similar for the two doses, which was probably related to the high level of resistant Enterobacteriaceae exhibited before treatment. Our results suggest that a single treatment with 2 or 10 mg/kg marbofloxacin exerts a moderate selective pressure on commensal Enterobacteriaceae in YBs and in VCs. A fivefold decrease of marbofloxacin regimen did not affect the selection of resistances among commensal bacteria.

This work aimed to investigate whether different microbial assemblies in flour may influence the microbiological and biochemical characteristics of traditional sourdough. To reach this purpose, members of lactic acid bacteria, enterobacteria, and yeasts were isolated from durum wheat flour. Secondly, the isolated microorganisms (Pediococcus pentosaceus, Saccharomyces cerevisiae, Pantoea agglomerans, and Escherichia hermannii) were inoculated in doughs prepared with irradiated flour (gamma rays at 10 kGy), so that eight different microbial assemblies were obtained. Two non-inoculated controls were prepared, one of which (C-IF) using irradiated flour and the other (C) using non-irradiated flour. As shown by plate counts, irradiation of flour caused total inactivation of yeasts and a decrease of all the other microbial populations. However, acidification occurred also in the dough C-IF, due to metabolic activity of P. pentosaceus that had survived irradiation. After six fermentations, P. pentosaceus was the dominant lactic acid bacterium species in all the sourdoughs produced with irradiated flour (IF). Yet, IF-based sourdoughs broadly differed from each other in terms of strains of P. pentosaceus, probably due to the different microorganisms initially inoculated. Quantitative and qualitative differences of free amino acids concentration were found among the sourdoughs, possibly because of different microbial communities. In addition, as shown by culture-independent analysis (16S metagenetics), irradiation of flour lowered and modified microbial diversity of sourdough ecosystem.

Fluorescent labeling has been an invaluable tool for the study of living organisms and bacterial species are no exception to this. Here we present and characterize the pUltra plasmids which express constitutively a fluorescent protein gene (GFP, RFP, YFP or CFP) from a strong synthetic promoter and are suitable for the fluorescent labeling of a broad range of Enterobacteria. The amount of expressed fluorophore from these genetic constructs is such, that the contours of the cells can be delineated on the basis of the fluorescent signal only. In addition, labeling through the pUltra plasmids can be used successfully for fluorescence and confocal microscopy while unambiguous distinction of cells labeled with different colors can be carried out efficiently by microscopy or flow cytometry. We compare the labeling provided by the pUltra plasmids with that of another plasmid series encoding fluorescent proteins and we show that the pUltra constructs are vastly superior in signal intensity and discrimination power without having any detectable growth rate effects for the bacterial population. We also use the pUltra plasmids to produce mixtures of differentially labeled pathogenic Escherichia, Shigella and Salmonella species which we test during infection of mammalian cells. We find that even inside the host cell, different strains can be distinguished effortlessly based on their fluorescence. We, therefore, conclude that the pUltra plasmids are a powerful labeling tool especially useful for complex biological experiments such as the visualization of ecosystems of different bacterial species or of enteric pathogens in contact with their hosts.

The objective of this study was to perform an inventory of the extended-spectrum-β-lactamase (ESBL)-producing Enterobacteriaceae isolates responsible for infections in French hospitals and to assess the mechanisms associated with ESBL diffusion. A total of 200 nonredundant ESBL-producing Enterobacteriaceae strains isolated from clinical samples were collected during a multicenter study performed in 18 representative French hospitals. Antibiotic resistance genes were identified by PCR and sequencing experiments. The clonal relatedness between isolates was investigated by the use of the DiversiLab system. ESBL-encoding plasmids were compared by PCR-based replicon typing and plasmid multilocus sequence typing. CTX-M-15, CTX-M-1, CTX-M-14, and SHV-12 were the most prevalent ESBLs (8% to 46.5%). The three CTX-M-type EBSLs were significantly observed in Escherichia coli (37.1%, 24.2%, and 21.8%, respectively), and CTX-M-15 was the predominant ESBL in Klebsiella pneumoniae (81.1%). SHV-12 was associated with ESBL-encoding Enterobacter cloacae strains (37.9%). qnrB , aac(6 ′ )-Ib-cr , and aac(3)-II genes were the main plasmid-mediated resistance genes, with prevalences ranging between 19.5% and 45% according to the ESBL results. Molecular typing did not identify wide clonal diffusion. Plasmid analysis suggested the diffusion of low numbers of ESBL-encoding plasmids, especially in K. pneumoniae and E. cloacae . However, the ESBL-encoding genes were observed in different plasmid replicons according to the bacterial species. The prevalences of ESBL subtypes differ according to the Enterobacteriaceae species. Plasmid spread is a key determinant of this epidemiology, and the link observed between the ESBL-encoding plasmids and the bacterial host explains the differences observed in the Enterobacteriaceae species.

This study was conducted to determine the presence of enterobacteria in the eggs of ostriches reared on a farm with a history of reproductive failure. Ninety samples from twenty eggs were submitted to bacteriological tests. The results showed Enterobacteria growth in 100% of the eggs. The microorganisms isolated were Hafnia alvei in 50% (10/20), Serratia spp. in 20% (4/20), Escherichia coli in 15% (3/20), and Citrobacter freundii in 15% (3/20). All eggs presented poor eggshell quality, which favored enterobacteria contamination. Hafnia alvei was present only in the internal egg structures (albumen and yolk sac), suggesting the possibility of vertical infection.

Horizontal gene transfer (HGT), non-hereditary transfer of genetic material between organisms, accounts for a significant proportion of the genetic variability in bacteria. In Gram negative bacteria, the nucleoid-associated protein H-NS silences unwanted expression of recently acquired foreign DNA. This, in turn, facilitates integration of the incoming genes into the regulatory networks of the recipient cell. Bacteria belonging to the family Enterobacteriaceae express an additional protein, the Hha protein that, by binding to H-NS, potentiates silencing of HGT DNA. We provide here an overview of Hha-like proteins, including their structure and function, as well as their evolutionary relationship. We finally present available information suggesting that, by expressing Hha-like proteins, bacteria such as Escherichia coli facilitate HGT incorporation and hence, the impact of HGT in their genetic diversity.

Foram avaliadas as populações microbianas, o pH, a relação nitrogênio amoniacal/nitrogênio total (N-NH3), as concentrações de ácidos láctico, acético e butírico e a composição bromatológica de silagens de Brachiaria decumbens cv. Basiliski de cinco idades de rebrotação (30, 40, 50, 60 e 70 dias) utilizando-se silos laboratoriais de 2 kg de capacidade. Utilizou-se um esquema fatorial 5 × 6 (5 idades de rebrotação × 6 períodos de fermentação), em um delineamento inteiramente casualizado, com três repetições. Os períodos de fermentação foram 1, 3, 7, 14, 28 e 56 dias. Na forragem fresca, foram registradas populações de bactérias do ácido láctico variando entre 3,93 (30 dias de rebrotação) e 5,51 (70 dias de rebrotação) log unidades formadoras de colônia (ufc)/g de forragem. Nas silagens, foram registradas populações máximas desses microrganismos no sétimo dia de fermentação (8,69 log ufc/g de silagem). As populações de enterobactérias persistiram até o 28º dia, com os valores máximos observados logo no primeiro dia de fermentação (7,89 log ufc/g de silagem). Os teores de MS, FDN, FDA e NIDA aumentaram linearmente, enquanto os de PB diminuíram linearmente com a idade de rebrotação das plantas. Houve redução linear dos teores de MS, PB e FDN e aumento linear dos teores de NIDA com o avanço do período fermentativo. O teor de N-NH3 decresceu e aumentou linearmente com a idade de rebrotação e com o período de fermentação, respectivamente. O pH decresceu exponencialmente com o período de fermentação. Os teores de ácido láctico aumentaram e os de ácido butírico decresceram linearmente com a idade de rebrotação. A espécie de Lactobacillus predominante em plantas de capim-braquiária é o Lactobacillus plantarum. Microbial populations, pH, ammonium nitrogen/total nitrogen (N-NH3) ratio, concentrations of lactic acid, acetic acid and butyric acid and the chemical composition of Brachiaria decumbens cv. Basiliski silages at different regrowth ages (30, 40, 50, 60 and 70 days) were evaluated by using 2-kg capacity laboratory silos. It was used a 5 × 6 factorial scheme (5 regrowth ages × 6 fermentation periods) in a complete random design, with three replicates. The fermentation periods were 1, 3, 7, 14, 28 and 56 days. Lactic acid bacteria populations were recorded in fresh forage varying from 3.93 (30 days of regrowth) to 5.51 (70 days of regrowth) log colony-forming units (cfu)/g forage. Maximum populations of these microorganisms were recorded in the silages on the seventh day of fermentation (8.69 log cfu/g silage). Enterobacteria populations persisted until the 28th day, with maximum values found as early as the first day of fermentation (7.89 log cfu/g silage). Levels of DM, NDF, ADF and ADIN increased linearly whereas values of CP decreased linearly with age of regrowth of the plants. There was a linear reduction in the levels of DM, CP and NDF and linear increase in the levels of ADIN over the period of fermentation. N-NH3 level decreased and increased linearly with regrowth age and fermentation period, respectively. The pH decreased exponentially with fermentation period. Lactic acid increased and butyric acid decreased linearly with regrowth age. The predominant Lactobacillus plantarum species in signalgrass plants is Lactobacillus plantarum.

It was studied, in this work, the effects of supplementation of probiotics on the performance, on the pH of crop content, on the morphometry of the ileum and on the number of enterobacteria in the cecum of the broilers. Out of the 120 studied birds, 60 belonged to the test group and 60 to the control group. At 1, 7, 12, 18, 23 and 28 days of age, the chickens and rations were weighed and mortality and morbidity were determined. Then, the birds were sacrificed and the pH of the crop content was measured, ileum fragments were collected for morphometric analysis and the enterobacteria in the cecal content were quantified. The mean pH was lower in the group that was given probiotics at 1, 7 and 18 days of age. There were no differences on ration consumption, weight, mortality and morbidity rates. The counts of enterobacteria were lower in the supplemented animals at the ages of 7, 18 and 28 days. The measure of villus of the ileum was higher for the group treated with probiotics at all ages, except for 1 day of age in comparison to the control group. This work proves that supplementation with probiotics reduces the pH of the crop content at the ages of 1, 7 and 18 days and it can contribute to the reduction of pathogenic bacteria on the first days of life by reducing the quantity of enterobacteria in the cecum. In addition, probiotics increase the height of villi in the ileum of the birds. Neste trabalho foram estudados os efeitos da suplementação de probióticos sobre o desempenho, o pH do conteúdo do papo, a morfometria do íleo e o número de enterobactérias no ceco de frangos de corte. Das 120 aves estudadas, 60 eram pertencentes ao grupo teste e 60 ao grupo controle. Nas idades de 1, 7, 12, 18, 23 e 28 dias, os frangos e a ração foram pesados e a mortalidade e morbidade foram determinadas. As aves foram, então sacrificadas, o pH do papo foi medido, os fragmentos do íleo foram coletados para análise da morfometria e as enterobactérias do ceco foram quantificadas. A média do pH foi menor no grupo que recebeu probióticos nas idades de 1, 7 e 18 dias. Não houve diferença no consumo de ração, no peso das aves, e nas taxas de mortalidade e morbidade. A contagem de enterobactérias foi menor nos animais sob suplementação com probióticos nas idades de 7, 18 e 28 dias. A medida das vilosidades do íleo foi maior no grupo tratado com probióticos em todas as idades, à exceção de 1 dia em relação ao grupo controle. Este trabalho comprova que a suplementação com probióticos diminui o pH do conteúdo do papo nas idades de 1, 7 e 18 dias, e pode contribuir na redução de bactérias patogênicas nos primeiros dias de vida, diminuindo a quantidade de enterobactérias no ceco. Além disso, os probióticos aumentaram a altura dos vilos do íleo das aves.