Colonisation by multidrug-resistant organisms in health workers in primary care: narrow spectrum oral antimicrobials are a risk factor.

BACKGROUND: Limited information exists on carriage of multidrug-resistant organisms (MDRO) by health workers (HWs) in primary care settings. This study aims to determine the prevalence of MDRO carriage among HWs in primary care and to identify associated risk factors. METHODS: A cross-sectional study was conducted across all 12 primary care units in São Caetano do Sul-SP, Brazil, from October to December 2023. Self-collected samples (nasal, oropharyngeal, and inguinal) were obtained. Environment cultures (potable water, sewage and stream water) were evaluated. Stenotrophomonas maltophilia isolates (human and environmental) were typed. RESULTS: The study included 265/288 (92%) of HWs in primary care teams, mostly women with a median age of 47 years (IQR 38-57); 78% had no comorbidities. MDRO colonisation was found in 8.7% (23 HWs). The following bacteria were found: S. maltophilia (n = 9; 3.4%) in inguinal swabs; methicillin-resistant Staphylococcus aureus (n = 8; 3%) from all sites; extended-spectrum ß-lactamase-producing bacteria (n = 5; 2%) in inguinal swabs; and vancomycin-resistant enterococci in an inguinal swab (n = 1; 0.4%). Previous antibiotic use was significantly associated with MDRO colonisation (OR 2.91, 95% CI 1.19-7.09, p = 0.018), mainly narrow spectrum oral beta-lactams and macrolides. S. malthophilia was polyclonal and human and environmental isolates differed. CONCLUSION: Colonisation by MRSA, VRE, and ESBL-producing bacteria was low; however, 4% were surprisingly colonized by polyclonal S. maltophilia. This pathogen may also suggest using narrow-spectrum rather than the expected broad-spectrum antimicrobials. Antibiotic use was the only risk factor found, mainly with oral narrow-spectrum drugs.

Early Candida colonisation impact on patients and healthcare professionals in an intensive care unit.

OBJECTIVES: Candida spp. is an opportunistic pathogen that causes superficial and invasive infections with nosocomial outbreaks without strict hygiene protocols. Herein, we assessed oral colonisation by Candida spp. in 209 Intensive Care Unit (ICU) patients between July 2021 and April 2022, conducting clinical, epidemiological, and microbiological characterisation of those developing oral or invasive candidiasis. METHODS: Initial oral swabs were collected within 24 h of admission in the ICU, followed by collections on Days 2, 4, 6 and 8. Swabs from denture-wearing patients, abiotic surfaces, healthcare professionals' hands, and retroauricular regions were also obtained. Recovered yeasts and filamentous fungi were identified using MALDI-TOF MS and morphological characteristics, respectively. Genetic similarity of Candida spp. isolates was evaluated using Amplified fragment length polymorphism (AFLP), and the antifungal susceptibility profile was determined by broth microdilution. RESULTS: In the study, 64.11% of patients were orally colonised by Candida spp. Of these, 80.59% were colonised within the first 24 h. Oral colonisation also occurred on subsequent days: 50%/Day 2, 26.92%/Day 4, and 11.53%/Days 6 and 8. Of the patients, 8.61% had oral candidiasis, mainly pseudomembranous. Among orally colonised patients, 2.23% developed invasive candidiasis. Besides, 89.47% of healthcare professionals evaluated were colonised. MALDI-TOF MS identified different yeast species, and C. albicans (45.34%), C. tropicalis (15.7%), and C. parapsilosis sensu stricto (9.88%) were the most prevalent. AFLP analysis indicated a high genetic correlation (≥97%) between C. parapsilosis sensu stricto isolates from patients and professionals. Three resistant C. albicans isolates were also found. CONCLUSION: This study reported a diversity of yeast and filamentous fungi species in ICU patients and highlighted early Candida spp. colonisation risks for invasive candidiasis, as well as the potential horizontal transmission in the nosocomial setting, emphasising the need for effective infection control measures.

Metagenomics reveals an abundance of black yeast-like fungi in the skin microbiome.

BACKGROUND: The skin is the first line of defence against communities of resident viruses, bacteria and fungi. The composition of the microbiome might change with factors related to the environment and host. The microbiome is dominated by bacteria. Dermatophytes and yeasts are the predominant fungi that are also involved in opportunistic infections of skin, hair and nails. Among environmental fungi, Chaetothyriales (black yeasts and relatives) are enriched by hydrocarbon pollution in domesticated habitats and comprise numerous species that cause mild-to-severe disease. METHODS: We investigated the presence of black fungi in the skin microbiome by conducting an analysis in the publicly available metagenomic SRA database (NCBI). We focused on the causative agents of chromoblastomycosis and phaeohyphomycosis and used barcodes and padlock probe sequences as diagnostic tools. RESULTS: A total of 132,159,577 MB was analysed and yielded 18,360 reads that matched with 24 species of black fungi. Exophiala was the most prevalent genus, and Cyphellophora europaea was the most abundant species. CONCLUSION: This study reveals the abundant presence of Chaetothyriales on the skin without necessarily being associated with infection. Most of the detected causal agents are known from mild skin diseases, while also species were revealed that had been reported from CARD9-deficient patients.

Carriage prevalence, serotype distribution, antimicrobial resistance, pspA typing and pilus islets of Streptococcus pneumoniae isolated from adults living in a Brazilian urban slum.

INTRODUCTION: For Brazilian adults, pneumococcal vaccines have been usually taken only by those who are at higher risk for development of pneumococcal diseases. Since populations from lower socioeconomic status are at high risk of acquiring pneumococcal infections, we investigated the carriage prevalence, colonization risk factors, capsular and surface protein types, and antimicrobial resistance among pneumococcal isolates recovered from adults living in a Brazilian urban slum. METHODS: Between September-December 2016, we conducted a cross-sectional study among individuals aged ≥ 18 years who attended a public primary clinic in Niterói/RJ, Brazil. Pneumococci were isolated by culture on sheep blood agar plates with and without gentamicin. Antimicrobial susceptibility was determined for all isolates. We used PCR to determine capsular types, PspA families (Fam) and pilus islets (PI). RESULTS: Of 385 adults, 32 (8.3 %) were pneumococcal carriers. Three carriers had two different pneumococci, totaling 35 isolates. After multivariate analysis, smoking, previous hospitalization, alcohol consumption and co-habitation with children aged < 6 years increased the odds of pneumococcal carriage, but antibiotic use in the previous 2 weeks was found to be a protective factor. Fourteen different serogroups/serotypes were detected and the prevalent ones were 9 N/L, 10A, 15B/C and 35F/47F (n = 3; 8.6 % each). Non-typeable (NT) isolates made up 31.4 %. All isolates were susceptible to chloramphenicol, levofloxacin and vancomycin. We found eight (22.9 %) penicillin non-susceptible pneumococci (PNSP) with minimum inhibitory concentrations (MICs) of 0.38-1.5 µg/mL. The two (5.7 %) erythromycin-resistant isolates had MIC > 256 µg/mL, cMLSB phenotype and the erm(B) gene. Twelve (34.3 %) and 17 (48.6 %) isolates had PspA Fam1 and Fam2, respectively. Three (8.6 %) isolates had genes for pilitwo PI-1 and one PI-2. CONCLUSION: We detected a low frequency of pneumococcal carriage among the adult population, but a high diversity of serotypes. Frequencies of PNSP and NT isolates resistant to antimicrobial agents are concerning.

A dimensão viva da experiência de supervisão didática / The living dimension of the didactic supervision experience / La dimensión viva de la experiencia de supervisión didáctica / La dimension vivante de l'expérience de supervision didactique

As autoras abordam uma experiência de supervisão, para fins de formação analítica, em um instituto de psicanálise, desenvolvida pela analista e pela supervisora implicadas na tarefa. Partindo de uma retrospectiva histórica da supervisão no tripé da formação psicanalítica, propõem uma reflexão sobre seu lugar e sua importância no exercício do vir a ser analista. O estudo leva as autoras a acreditarem que, assim como a própria análise é uma tarefa interminável, as forças intrapsíquicas que estão permanentemente em jogo no encontro do analista com o paciente geram uma demanda que convoca o diálogo na supervisão, o qual parece conter o paradoxo do descobrimento sem colonização. Em conclusão, entendem que, ao mesmo tempo que a tarefa da supervisão promove novos significados, ela não deve se tornar um exercício de doutrinamento.

supervisão didática

formação psicanalítica

história da supervisão

descobrimento sem colonização

This paper regards a supervisory experience for analytical specialization purposes in an Institute of Psychoanalysis, developed by the analyst and supervisor involved in the task. The authors, based on an analysis of the historical dimension of supervision on the tripod of psychoanalytic training, propose a reflection on the supervision's role in the process of becoming an analyst. The study leads the authors to believe that, just as the analysis itself is an endless task, the intrapsychic forces that are permanently at stake in the encounter of the analyst with the patient generate a demand that calls for the dialogue in the supervision, which seems to contain the paradox of discovery without colonization. In conclusion, the authors consider that, while the task of supervision promotes new meanings, it should not become an exercise of indoctrination.

El trabajo trata de una experiencia de supervisión, con fines de formación analítica en un Instituto de Psicoanálisis, desarrollada por el analista y el supervisor involucrado en la tarea. Los autores, partiendo de una retrospectiva histórica de la supervisión sobre el trípode de la formación psicoanalítica, proponen una reflexión sobre su lugar e importancia en el ejercicio de convertirse en analista. El estudio lleva a los autores a creer que, así como el análisis en sí es una tarea sin fin, las fuerzas intrapsíquicas que están permanentemente en juego en el encuentro del analista con el paciente generan una demanda que llama al diálogo en la supervisión, que parece contener la paradoja de descubrimiento sin colonización. En conclusión, se entiende que, si bien la tarea de supervisión promueve nuevos significados, no debe convertirse en un ejercicio de adoctrinamiento.

L'article aborde une expérience de supervision, à des fins de la formation analytique dans un Institut de Psychanalyse, développée par une analyste et une superviseuse qui sont impliquées dans la tâche. Les autrices, à partir d'une rétrospective historique de la supervision sur le trépied de la formation psychanalytique, proposent une réflexion sur leur place et leur importance dans l'exercice de devenir analyste. L'étude conduit les autrices à croire que, de la même manière que l'analyse elle-même est une tâche sans fin, les forces intrapsychiques, qui sont en permanence en jeu dans la rencontre de l'analyste avec le patient, génèrent une demande qui appelle au dialogue dans la supervision, lequel semble contenir le paradoxe de la découverte sans colonisation. En conclusion, on comprend que, lors même que la supervision promeuve de nouvelles significations, elle ne doive pas devenir un exercice d'endoctrinement.

Ancestral genetic legacy of the extant population of Argentina as predicted by autosomal and X-chromosomal DIPs.

Aiming to determine their ancestry diagnostic potential, we selected two sets of nuclear deletion/insertion polymorphisms (DIPs), including 30 located on autosomal chromosomes and 33 on the X chromosome. We analysed over 200 unrelated Argentinean individuals living in urban areas of Argentina. As in most American countries, the extant Argentinean population is the result of tricontinental genetic admixture. The peopling process within the continent was characterised by mating bias involving Native American and enslaved African females and European males. Differential results were detected between autosomal DIPs and X-DIPs. The former showed that the European component was the largest (77.8%), followed by the Native American (17.9%) and African (4.2%) components, in good agreement with the previously published results. In contrast, X-DIPs showed that the European genetic contribution was also predominant but much smaller (52.9%) and considerably larger Native American and African contributions (39.6% and 7.5%, respectively). Genetic analysis revealed continental genetic contributions whose associated phenotypic traits have been mostly lost. The observed differences between the estimated continental genetic contribution proportions based on autosomal DIPs and X-DIPs reflect the effects of autosome and X-chromosome transmission behaviour and their different recombination patterns. This work shows the ability of the tested DIP panels to infer ancestry and confirm mating bias. To the best of our knowledge, this is the first study focusing on ancestry-informative autosomal DIP and X-DIP comparisons performed in a sample representing the entire Argentinean population.

Comparative Genomics Provides Insights into the Taxonomy of Azoarcus and Reveals Separate Origins of Nif Genes in the Proposed Azoarcus and Aromatoleum Genera.

Among other attributes, the Betaproteobacterial genus Azoarcus has biotechnological importance for plant growth-promotion and remediation of petroleum waste-polluted water and soils. It comprises at least two phylogenetically distinct groups. The "plant-associated" group includes strains that are isolated from the rhizosphere or root interior of the C4 plant Kallar Grass, but also strains from soil and/or water; all are considered to be obligate aerobes and all are diazotrophic. The other group (now partly incorporated into the new genus Aromatoleum) comprises a diverse range of species and strains that live in water or soil that is contaminated with petroleum and/or aromatic compounds; all are facultative or obligate anaerobes. Some are diazotrophs. A comparative genome analysis of 32 genomes from 30 Azoarcus-Aromatoleum strains was performed in order to delineate generic boundaries more precisely than the single gene, 16S rRNA, that has been commonly used in bacterial taxonomy. The origin of diazotrophy in Azoarcus-Aromatoleum was also investigated by comparing full-length sequences of nif genes, and by physiological measurements of nitrogenase activity using the acetylene reduction assay. Based on average nucleotide identity (ANI) and whole genome analyses, three major groups could be discerned: (i) Azoarcus comprising Az. communis, Az. indigens and Az. olearius, and two unnamed species complexes, (ii) Aromatoleum Group 1 comprising Ar. anaerobium, Ar. aromaticum, Ar. bremense, and Ar. buckelii, and (iii) Aromatoleum Group 2 comprising Ar. diolicum, Ar. evansii, Ar. petrolei, Ar. toluclasticum, Ar. tolulyticum, Ar. toluolicum, and Ar. toluvorans. Single strain lineages such as Azoarcus sp. KH32C, Az. pumilus, and Az. taiwanensis were also revealed. Full length sequences of nif-cluster genes revealed two groups of diazotrophs in Azoarcus-Aromatoleum with nif being derived from Dechloromonas in Azoarcus sensu stricto (and two Thauera strains) and from Azospira in Aromatoleum Group 2. Diazotrophy was confirmed in several strains, and for the first time in Az. communis LMG5514, Azoarcus sp. TTM-91 and Ar. toluolicum TT. In terms of ecology, with the exception of a few plant-associated strains in Azoarcus (s.s.), across the group, most strains/species are found in soil and water (often contaminated with petroleum or related aromatic compounds), sewage sludge, and seawater. The possession of nar, nap, nir, nor, and nos genes by most Azoarcus-Aromatoleum strains suggests that they have the potential to derive energy through anaerobic nitrate respiration, so this ability cannot be usefully used as a phenotypic marker to distinguish genera. However, the possession of bzd genes indicating the ability to degrade benzoate anaerobically plus the type of diazotrophy (aerobic vs. anaerobic) could, after confirmation of their functionality, be considered as distinguishing phenotypes in any new generic delineations. The taxonomy of the Azoarcus-Aromatoleum group should be revisited; retaining the generic name Azoarcus for its entirety, or creating additional genera are both possible outcomes.

Prevalence of oxacillin-susceptible methicillin-resistant Staphylococcus aureus nasal carriage and their clonal diversity among patients attending public health-care facilities.

Context: Nosocomial infections arise from many microorganisms, including Staphylococcus aureus. Aims: The aim of this study is to determine the molecular epidemiology of circulating methicillin-resistant S. aureus (MRSA) clones among patients attending community and health-care facilities in Nova Friburgo, RJ, Brazil. Methods: A total of 1002 nasal swab samples were collected from May 2010 to September 2015. S. aureus isolates were identified through phenotypic tests, submitted to antimicrobial susceptibility tests and genotypic analysis to detect mecA, panton-valentine leucocidin (PVL) genes, SCCmec, SPA and multilocus sequencing typing (MLST) typing. Results: We identified 294 (29.3%) isolates as S. aureus and 91 (9.1%) as MRSA. A total of 17 isolates did not present a correlation between phenotypic and genotypic resistance profiles. Among MRSA isolates, 17 (18.7%) carried PVL genes. A total of 20 different SPA types were determined, being grouped by MLST into eight different sequence types. ST5/t002 was the most prevalent genotype found among these isolates. Conclusions: There is a gradual colonisation shift happening in the infection pattern by S. aureus in Brazil. The Brazilian Epidemic Clone (ST239-SCCmec IIIa-PVL-) seems to be substituted by isolates from different clonal complexes, such as ST5, ST8 and ST30. The non-correlation between phenotypic/genotypic resistance profile observed in some isolates suggests the presence of other methicillin resistance mechanisms different from mecA presence or a difference in the nucleotide sequence, which prevents the primers to identify the specific region during polymerase chain reaction reactions. MRSA identification should be based on phenotypic and genotypic testing to ensure the various types of resistance mechanisms.

Bacterial Colonisation of Suction Cannulas in Orthopaedic Surgery. / Colonización bacteriana de las cánulas de succión en cirugía ortopédica.

INTRODUCTION: Osteoarticular infections represent a major complication in orthopaedic surgery. The aim is to identify the percentage of suction cannulas colonised and to determine the relationship between the time they are used in surgery and the colonisation of these cannulas. MATERIALS AND METHODS: Descriptive and prospective study that analysed 546 suction cannulas used in clean orthopaedic surgery in a trauma centre, between November 2017 and March 2018. The distal end of the cannula was cultured to determine the colonisation rate. RESULTS: 7.3% of the cultured cannulas were positive for pathogens, the most frequent being Staphylococcus epidermidis at 27.5%. In addition, an association was found between colonisation and the length of time the cannula was used. The possibility of colonisation of cannulas used for between 60minutes or more, is greater than those used for less than 60minutes; between 60 and 90minutes the possibility is twice as high OR= 2.2 (CI:95% 1.1 - 4.1) and in cannulas used for more than 90minutes it is 8 times higher OR= 8.49 (CI:95% 1.77 - 40.86). CONCLUSIONS: The colonisation rate of cannulas is lower than reported in the literature. The longer the cannula is used in surgery increases the risk of their colonisation. Follow-up studies are being considered to determine whether suction cannula colonisation is associated with increased postoperative infection.

¿Colonización de subjetividades en el sub-desarrollo? / Colonization of subjectivities in sub-development? / Colonização de subjetividades em subdesenvolvimento? / ¿Colonisation des subjectivités dans le sous-développement?

Nuestros países presentan singularidades contradictorias y complejas que devienen en un conjunto de enfoques y acciones hacia la modernidad en la invención del sujeto llamado Ciudadano. Esa construcción simbólica del sujeto, produce subjetividades que hacen a la institucionalización de ciertas prácticas sociales, políticas, culturales. El ciudadano se definió en lógicas heterónomas que lo constituyen como tal y, en ese sentido, construir gobernabilidad ha sido un constante problema de nuestras sociedades, transitando procesos no siempre dialécticos entre la búsqueda casi frenética por el progreso y el desarrollo, con un desprendimiento del pasado identificado con el atraso y la barbarie. Pretendemos en este trabajo reflexionar sobre esa idea de desarrollo, sus implicaciones para la Independencia dependiente y las producciones de subjetividad colonizadas, desde conceptos sustantivos de la Psicología de la Liberación de Martín-Baró como la naturalización y la ideologización.

Our countries have become singularities as a set of approaches and actions towards modernity contradictory and complex in the invention of the subject called “Citizen”. We think that this symbolic construction of the subject, produces specific subjectivities that make the institutionalization of social practices, political, cultural. The “citizen” was defined in heteronomous logics that constitute as such and, in that sense, build governance has been a problem in our societies dialectical processes are not always traveling between almost frantic search for “progress” and “development” with detachment of the past, identified with backwardness and “barbarism”. We aim in this work reflect on the idea of development, its implications for Independence Productions dependent and colonized subjectivity, from substantive concepts of the Psychology of Liberation of Martín-Baró as naturalization and ideologization.

Nossos países apresentam singularidades contraditórias e complexas que se tornam um conjunto de abordagens e ações em direção à modernidade na invenção do sujeito chamado Cidadão. Essa construção simbólica do sujeito produz subjetividades que fazem a institucionalização de certas práticas sociais, políticas e culturais.O "cidadão" foi definido em lógicas heteronomas que constituem, como tal, e, nesse sentido, a construção de uma governação tem sido um problema em nossas sociedades, em trânsito de processos não sempre dialéticos, viajando entre a busca quase frenética de "progresso" e "desenvolvimento" com desapego do passado, identificado com atraso e "barbárie". Nosso objetivo neste trabalho e refletir sobre a idéia de desenvolvimento, suas implicações para a independência dependente e as produções de subjetividades colonizadas, a partir de conceitos substantivos da Psicologia da Libertação de Martín-Baró como naturalização e ideologização.

Nos pays présentent des singularités complexes et contradictoires qui deviennent un ensemble d'approches et d'actions vers la modernité dans l'invention du sujet appelé Citizen. Cette construction symbolique du sujet produit des subjectivités que font-ils a l'institutionnalisation de certaines pratiques sociales, politiques et culturelles. Le citoyen s'est défini dans des logiques hétéronomes qui le constituent en tant que tel et, en ce sens, la construction de la gouvernance a été un problème constant de nos sociétés, en transit processus pas toujours dialectiques entre la recherche presque frénétique du progrès et le développement, avec un détachement du passé, identifié avec le retard et la barbarie. Dans cet article, nous avons l'intention de réfléchir à cette idée de développement, à ses implications pour les productions d'indépendance-dépendante et de subjectivité colonisée, à partir des concepts de fond de la psychologie de la libération de Martín-Baró comme la naturalisation et l' idéologisation.

Salt stress and hydroxyectoine enhance phosphate solubilisation and plant colonisation capacity of Kosakonia radicincitans.

Gram-negative bacterial endophytes have attracted research interest caused by their advantageous over epiphytic bacteria in plant nutrition and protection. However, research on these typically Gram-negative endophytes has deficiencies concerning the role of cultivation and pre-formulation strategies on further plant colonisation capabilities. Besides, the influence of cultivation conditions and osmotic stress within bacterial endophytes on their phosphate solubilising ability has not yet been addressed. By pre-conditioning cells with an osmoadaptation and a hydroxyectoine accumulation approach, this research aimed at enhancing the capability of the plant growth promoting bacterium Kosakonia radicincitans strain DSM 16656T to both solubilise phosphate and colonise plant seedlings. The results showed that halotolerant bacterial phenotypes increased the root-colonising capability by approximately 3-fold and presented growth-promoting effects in radish plants. Interestingly, findings also demonstrated that salt stress in the culture media along with the accumulation of hydroxyectoine led to an increase in the in vitro phosphate-solubilising ability by affecting the production of acid phosphatases, from 1.24 to 3.34 U mg-1 for non-salt stressed cells and hydroxyectoine-added cells respectively. Thus, this approach provides a useful knowledge upon which the salt stress and compatible solutes in bacteria endophytes can confer phenotypic adaptations to support the eco-physiological performance concerning phosphate-solubilising abilities and endosphere establishment.

Shredder Chironomid Diets Are Influenced by Decomposition Rates of Different Leaf Litter Species.

The diet of shredder chironomid larvae depends on the local and temporal conditions of the food resources. We analysed the gut content of shredder chironomid larvae that colonised the leaf litter of three riparian species: Hedychium coronarium, Pteridium arachnoideum and Magnolia ovata. We hypothesised that the differences in the decomposition rates of leaf litter species influence the consumption of plant tissue by shredder chironomid taxa over time. We incubated perforated bottles with each leaf species within four low-order streams during 1st, 3rd, 7th, 22nd, 36th, 55th and 85th day of exposure. We used an analysis of covariance (ANCOVA) to compare differences in the percentage of AFDM (ash-free dry mass) and AOM (amorphous organic matter) among leaf litter species. To verify differences in the larvae abundance, we used a general linear model, and to test if there were feeding preferences for AFDM and AOM, we used the adapted Paloheimo selectivity index. Magnolia ovata presented a higher quantity of AOM followed by H. coronarium and P. arachnoideum. Pteridium arachnoideum showed a higher AFDM followed by H. coronarium and M. ovata. The larvae abundance was different among plant species and varied significantly with AFDM and AOM quantities. The consumption of plant tissue by shredder chironomid differed temporarily and among riparian species, where facultative or strict shredders showed strong association with different leaf litter species. The amount of AFDM and AOM in plant tissues explained these differences. We highlighted that shredder chironomids displayed an important role as co-participants in the decomposition process.

Maternal antibiotic prophylaxis affects Bifidobacterium spp. counts in the human milk, during the first week after delivery.

Human milk is an important source of microorganisms for infant gut colonisation. Although the maternal antibiotic prophylaxis is an important strategy to prevent maternal/neonatal sepsis, it has to be investigated how it may affect the human milk microbiota, especially the genus Bifidobacterium, which has been associated to health benefits. Here, we investigated the impact of the maternal antibiotic prophylaxis on the human milk Bifidobacterium spp. and total bacteria counts, in the first week (short-term) and first month (medium-term) after delivery. Human milk samples were collected from 55 healthy lactating women recruited from the University Hospital of the University of São Paulo at days 7±3 and 30±4 after vaginal delivery. Twenty one volunteers had received maternal antibiotic prophylaxis (MAP group) and 34 had not received MAP (no-MAP group) during or after labour. Total DNA was isolated from milk samples, and the bacterial counts were estimated by quantitative PCR (qPCR). We found lower levels of Bifidobacterium in the MAP group in the first week after delivery (median = 2.1 vs 2.4 log of equivalent cells/ml of human milk, for MAP and no-MAP groups, respectively; P=0.01), although there were no statistical differences in total bacteria count. However, no differences were found in Bifidobacterium counts between the groups at day 30±4 (median = 2.5 vs 2.2 log of equivalent cells/ml of human milk, for MAP and no-MAP groups, respectively; P=0.50). Our results suggest that MAP has a significant impact on Bifidobacterium counts in human milk, reducing this population in the first week after delivery. However, throughout the first month after delivery, the Bifidobacterium counts tend to recover, reaching similar counts to those found in no-MAP group at day 30±4 after delivery.

High rates of methicillin-resistant Staphylococcus aureus colonisation in a Brazilian Intensive Care Unit.

OBJECTIVE: To evaluate the colonisation rate of Staphylococcus aureus in the oropharynx and the insertion site of central venous catheters in intensive care unit patients. DESIGN: Cross-sectional study. SETTING: Brazilian intensive care unit. MEASUREMENTS: Samples were collected from October to December 2015 from the oropharyngeal cavity and central venous catheter insertion site of 110 patients. Samples that presented growth of Staphylococcus aureus were isolated and their sensitivity profiles were tested for several antimicrobials. FINDINGS: The study participants (110) were mostly females older than 60 years (53.6%). The mean length of hospitalisation was 15.5 days (±31.3). A total of 188 biological samples were collected: 110 collected from the oropharynx and 78 from the central venous catheter insertion site. A 35% (n = 38/110) S. aureus colonisation rate of the patients was observed in at least one collection site. In the oropharynx alone, a 31% rate (n = 34/110) was found, and a 12.8% rate (n = 10/78) at central venous catheter insertion sites only. MRSA colonisation in the oropharynx or at the central venous catheter occurred in 29 (26.4%) patients and vancomycin resistant Staphylococcus aureus was present in 24 (22.4%) of the patients studied. Patients hospitalised for seven days or more were 4.8 times more likely to be colonised compared to patients hospitalised less than seven days (95% CI = 1.2-28.5). CONCLUSION: The oropharynx and the central venous catheter are important reservoirs of this bacterium that in critical conditions may become pathogenic. The data showed a high degree of resistance of the bacterial populations isolated to different drugs, which may hinder the control of these organisms.

Contribution of flagella and motility to gut colonisation and pathogenicity of Salmonella Enteritidis in the chicken

ABSTRACT Salmonella Enteritidis causes fowl paratyphoid in poultry and is frequently associated to outbreaks of food-borne diseases in humans. The role of flagella and flagella-mediated motility into host-pathogen interplay is not fully understood and requires further investigation. In this study, one-day-old chickens were challenged orally with a wild-type strain Salmonella Enteritidis, a non-motile but fully flagellated (SE motB) or non-flagellated (SE fliC) strain to evaluate their ability to colonise the intestine and spread systemically and also of eliciting gross and histopathological changes. SE motB and SE fliC were recovered in significantly lower numbers from caecal contents in comparison with Salmonella Enteritidis at early stages of infection (3 and 5 dpi). The SE motB strain, which synthesises paralysed flagella, showed poorer intestinal colonisation ability than the non-flagellated SE fliC. Histopathological analyses demonstrated that the flagellated strains induced more intense lymphoid reactivity in liver, ileum and caeca. Thus, in the present study the flagellar structure and motility seemed to play a role in the early stages of the intestinal colonisation by Salmonella Enteritidis in the chicken.(AU)

Contribution of flagella and motility to gut colonisation and pathogenicity of Salmonella Enteritidis in the chicken

ABSTRACT Salmonella Enteritidis causes fowl paratyphoid in poultry and is frequently associated to outbreaks of food-borne diseases in humans. The role of flagella and flagella-mediated motility into host-pathogen interplay is not fully understood and requires further investigation. In this study, one-day-old chickens were challenged orally with a wild-type strain Salmonella Enteritidis, a non-motile but fully flagellated (SE ΔmotB) or non-flagellated (SE ΔfliC) strain to evaluate their ability to colonise the intestine and spread systemically and also of eliciting gross and histopathological changes. SE ΔmotB and SE ΔfliC were recovered in significantly lower numbers from caecal contents in comparison with Salmonella Enteritidis at early stages of infection (3 and 5 dpi). The SE ΔmotB strain, which synthesises paralysed flagella, showed poorer intestinal colonisation ability than the non-flagellated SE ΔfliC. Histopathological analyses demonstrated that the flagellated strains induced more intense lymphoid reactivity in liver, ileum and caeca. Thus, in the present study the flagellar structure and motility seemed to play a role in the early stages of the intestinal colonisation by Salmonella Enteritidis in the chicken.

Milk fermented by Lactobacillus species from Brazilian artisanal cheese protect germ-free-mice against Salmonella Typhimurium infection.

Ingestion of milks fermented by Lactobacillus strains showing probiotic properties is an important tool to maintain gastrointestinal health. In this study, Lactobacillus rhamnosus D1 and Lactobacillus plantarum B7, isolated from Brazilian artisanal cheese, were used as starters for the functional fermented milks to assess their probiotic properties in a gnotobiotic animal model. Male germ-free Swiss mice received a single oral dose of milk fermented by each sample, and were challenged with Salmonella Typhimurium five days afterwards. Milk fermented by both Lactobacillus strains maintained counts above 108 cfu/ml during cold storage. Lactobacillus strains colonised the gut of the germ-free-mice, maintaining their antagonistic effect. This colonisation led to a protective effect against Salmonella challenge, as demonstrated by reduced pathogen translocation and histological lesions, when compared to control group, especially for Lactobacillus rhamnosus D1. Additionally, mRNA expression of inflammatory (interferon gamma, interleukin (IL)-6, tumour necrosis factor alpha) and anti-inflammatory (transforming growth factor ß1) cytokines was augmented in animals previously colonised and then challenged, when compared to other experimental groups. Lactobacillus plantarum B7 colonisation also promoted higher expression of IL-17, showing a proper maturation of colonised germ-free-mice immune system. IL-5 was stimulated by both strains' colonisation and not by S. Typhimurium challenge.

Contribution of flagella and motility to gut colonisation and pathogenicity of Salmonella Enteritidis in the chicken.

Salmonella Enteritidis causes fowl paratyphoid in poultry and is frequently associated to outbreaks of food-borne diseases in humans. The role of flagella and flagella-mediated motility into host-pathogen interplay is not fully understood and requires further investigation. In this study, one-day-old chickens were challenged orally with a wild-type strain Salmonella Enteritidis, a non-motile but fully flagellated (SE ΔmotB) or non-flagellated (SE ΔfliC) strain to evaluate their ability to colonise the intestine and spread systemically and also of eliciting gross and histopathological changes. SE ΔmotB and SE ΔfliC were recovered in significantly lower numbers from caecal contents in comparison with Salmonella Enteritidis at early stages of infection (3 and 5dpi). The SE ΔmotB strain, which synthesises paralysed flagella, showed poorer intestinal colonisation ability than the non-flagellated SE ΔfliC. Histopathological analyses demonstrated that the flagellated strains induced more intense lymphoid reactivity in liver, ileum and caeca. Thus, in the present study the flagellar structure and motility seemed to play a role in the early stages of the intestinal colonisation by Salmonella Enteritidis in the chicken.

Pneumococcal nasopharyngeal carriage among children in Brazil prior to the introduction of the 10-valent conjugate vaccine: a culture- and PCR-based survey.

We performed two different approaches (broth enrichment step prior to culture (BEC) and PCR (BEPCR)) for detecting Streptococcus pneumoniae from nasopharyngeal specimens collected from 242 children aged <6 years attending one hospital (n = 140) and one childcare centre (n = 102) in a major urban area in Brazil. These specimens were collected immediately before the introduction of the 10-valent pneumococcal conjugate vaccine (PCV10) and the 13-valent vaccine (PCV13) for routine use in Brazil. Results were compared with previous findings obtained with direct culture (DC) on a selective medium. Colonisation prevalence was 58·3% (n = 141), being higher among children attending the childcare centre (62·7% vs. 55%). The culture-based methods (DC and BEC) enabled the detection of S. pneumoniae in 119 (49·2%) and 115 (47·5%) children, respectively. The PCR-based method (BEPCR) was more sensitive and 137 (56·6%) carriers were identified. Twenty-six serogroups/serotypes were identified, predominantly 6B, 19F, 14, 6A, 15C and 23F. Multiple colonisation was observed in 13 (5·4%) children. The estimated serotypes coverage of available PCVs was 40·4% for the 10-valent (included in the Brazilian immunisation programme) and 55·8% for the 13-valent (only available in private clinics). The use of robust approaches to obtain a more realistic insight about the asymptomatic carrier status is of paramount importance to estimate and assess the impact of vaccine implementation. The combination between culture-based and molecular methods constitutes a suitable strategy.

Evaluation of colonisation resistance in stool of human donors using ex vivo, in vitro and in vivo assays.

The indigenous microbiota is the population of microorganisms normally present on the surface and mucosa of an individual, where it performs essential health functions, including the colonisation resistance (CR) against pathogens. To identify the bacteria responsible and the mechanisms involved in the CR, the germ-free (GF) animal model has been used, because in vitro studies cannot always be extrapolated to what occurs in vivo. In this study, ex vivo antagonism assays against seven enteropathogenic bacteria using stools from 15 healthy human donors confirmed that the CR showed individual variation. Using in vitro antagonism assays, 14 strains isolated from dominant faecal microbiota of donors with elevated CR were selected for mono-association in GF mice to test the in vivo antagonism against Salmonella enterica ser. Typhimurium. Mice mono-associated with Enterococcus hirae strain 8.2, Bacteroides thetaiotaomicron strain 16.2 and Lactobacillus ruminis strain 18.1 had significant reductions in faecal counts of the pathogen during the challenge. After five days of infection, the group associated with E. hirae 8.2 showed a reduction in the translocation of S. Typhimurium to the spleen, while the group associated with L. ruminis 18.1 presented an increased translocation to the liver. The histological data confirmed these results and revealed that the mice associated with E. hirae 8.2 showed fewer lesions on ileum and liver, compared to the damage caused by S. Typhimurium alone, while in mice associated with L. ruminis 18.1 there was significantly worse lesions. Concluding, from the dominant faecal microbiota from healthy human with high CR, through ex vivo, in vitro and in vivo assays, a bacterium was characterised for its high CR potential, being a candidate for probiotic use.