Rhinovirus infection and co-infection in children with severe acute respiratory infection during the COVID-19 pandemic period.

Rhinovirus causes respiratory tract infections in children and is found in co-infections. The objective of this research was to study the clinical profile of rhinovirus infection and co-infection in children with severe acute respiratory infection (SARI) during the COVID-19 pandemic period. We included 606 children ranging in age from 0.1 to 144 months of age from March 2020 to December 2021, hospitalized in the Pediatric Intensive Care Unit (PICU). The samples were collected by secretion from the nasopharynx region. A total of 259 children were tested positive for viral infection, 153 (59.07%) of them had a single rhinovirus infection and, 56 (36.6%) were aged between 60.1 and 144 months. Nine types of co-infections were identified and were found coinfection with three or more viruses (22/104, 21.15%). Observing the seasonality, the number of cases was similar between 2020 (49.53%) and 2021 (51.47%). Patients with a single infection (86.88%) and coinfection (67.30%) were more likely to have coughed. Patients with co-infection required the use of O2 for longer than those with a single rhinovirus infection. Hemogram results obtained from individuals with a single infection had higher levels of urea when compared to patients with co-infection with and other respiratory viruses. Multiple correspondence analyses indicated different clinical symptoms and comorbidities in patients with co-infection compared to those with single infection. The results found that the rhinovirus was much prevalent virus during the pandemic period and was found in co-infection with other virus types, what is important to diagnostic for the correct treatment of patients.

Identification of 2,4-diacetylphloroglucinol production in the genus Chromobacterium.

The compound 2,4-diacetylphloroglucinol (DAPG) is a broad-spectrum antibiotic that is primarily produced by Pseudomonas spp. DAPG plays an important role in the biocontrol disease suppressing activity of Pseudomonas spp. In the current study, we report the discovery of the DAPG biosynthetic cluster in strains of Chromobacterium vaccinii isolated from Brazilian aquatic environments and the distribution of the biosynthetic cluster in the Chromobacterium genus. Phylogenetic analysis of the phlD protein suggests the biosynthetic cluster probably entered the genus of Chromobacterium after a horizontal gene transfer event with a member of the Pseudomonas fluorescens group. We were able to detect trace amounts of DAPG in wild type cultures and confirm the function of the cluster with heterologous expression in Escherichia coli. In addition, we identified and verified the presence of other secondary metabolites in these strains. We also confirmed the ability of C. vaccinii strains to produce bioactive pigment violacein and bioactive cyclic depsipeptide FR900359. Both compounds have been reported to have antimicrobial and insecticidal activities. These compounds suggest strains of C. vaccinii should be further explored for their potential as biocontrol agents.

Profile analysis of emerging respiratory virus in children.

Acute respiratory infections (ARIs) are caused by a variety of microorganisms. Of all ARIs, 80% are caused by viruses such as human respiratory syncytial virus, metapneumovirus, influenza, parainfluenza, rhinovirus, and, more recently, Sars-CoV-2, which has been responsible for the COVID-19 pandemic. The objective of our study was to evaluate clinical data from a viral panel performed in children hospitalized with SARS or COVID-19 in the infirmary or ICU of 5 pediatric hospitals in the city of Goiânia, Goiás, Brazil. Demographic, clinical, and laboratory data were collected for analysis, and data on the outcomes underwent statistical treatment. A total of 128 patients were selected for the study, 54% of whom were male and 46% female. The viral panel included rhinovirus, COVID-19, metapneumovirus, adenovirus, and parainfluenza. Descriptive analyses of age profile showed differences in the involvement of particular viruses. The percentage of patients who required hospitalization in the ICU, infirmary, as well as individuals who were discharged after therapy or who died, were described. Our work shows that epidemiological surveillance measures are indispensable, especially if used in the continued analysis of viral panels in all pediatric patients with SARS.

Viral Coinfection of Children Hospitalized with Severe Acute Respiratory Infections during COVID-19 Pandemic.

The main pathogens of severe respiratory infection in children are respiratory viruses, and the current molecular technology allows for a rapid and simultaneous detection of a wide spectrum of these viral pathogens, facilitating the diagnosis and evaluation of viral coinfection. METHODS: This study was conducted between March 2020 and December 2021. All children admitted to the ICU with a diagnosis of SARI and who were tested by polymerase chain reaction on nasopharyngeal swabs for SARS-CoV-2 and other common respiratory viral pathogens were included in the study. RESULTS: The result of the viral panel identified 446 children, with one infected with a single virus and 160 co-infected with two or more viruses. This study employed descriptive analyses, where a total of twenty-two coinfections among SARI-causing viruses were identified. Thus, the five most frequent coinfections that were selected for the study are: hRV/SARS-CoV-2 (17.91%), hRV/RSV (14.18%), RSV/SARS-CoV-2 (12.69%), hRV/BoV (10.45%), and hRV/AdV (8.21%). The most significant age group was 38.1%, representing patients aged between 24 and 59 months (61 individuals). Patients older than 59 months represented a total of 27.5%, comprising forty-four patients. The use of oxygen therapy was statistically significant in coinfections with Bocavirus, other CoVs, Metapneumovirus, and RSV. Coinfections with SARS-CoV-2 and the other different coinfections presented a similar time of use of oxygen therapy with a value of (p > 0.05). In the year 2020, hRV/BoV was more frequent in relation to other types of coinfections, representing a total of 35.1%. The year 2021 presented a divergent profile, with hRV/SARS-CoV-2 coinfection being the most frequent (30.8%), followed by hRV/RSV (28.2%). Additionally, 25.6% and 15.4% represented coinfections between RSV/SARS-CoV-2 and hRV/AdV, respectively. We saw that two of the patients coinfected with hRV/SARS-CoV-2 died, representing 9.52% of all deaths in the study. In addition, both hRV/hBoV and hRV/RSV had death records for each case, representing 8.33% and 6.67% of all deaths, respectively. CONCLUSION: Coinfections with respiratory viruses, such as RSV and hBoV, can increase the severity of the disease in children with SARI who are admitted to the ICU, and children infected with SARS-CoV-2 have their clinical condition worsened when they have comorbidities.

Detection of Oxacillin/Cefoxitin Resistance in Staphylococcus aureus Present in Recurrent Tonsillitis.

BACKGROUND: Recurrent tonsillitis is one of the most common diseases in childhood, caused many times by ß-lactam-resistant S. aureus. The objective of this study was to investigate an alternative method to identify resistance to oxacillin/cefoxitin in S. aureus from hospitalized children with recurrent tonsillitis. METHODS: The samples of S. aureus came from patients with recurrent tonsillitis and were used in 16S rRNA sequencing and an antibiogram test for identification and verifying resistance, after which HSI methodology were applied for separation of S. aureus resistances. RESULTS: The S. aureus isolated showed sensitivity to oxacillin/cefoxitin and the diagnostic images show a visual description of the resistance different groups formed, that may be related to sensitivity and resistance to oxacillin/cefoxitin, characterizing the MRSA S. aureus. CONCLUSIONS: Samples that showed phenotypic resistance to oxacillin/cefoxitin were clearly separated from samples that did not show this resistance. A PLS-DA model predicted the presence of resistance to oxacillin/cefoxitin in S. aureus samples and it was possible to observe the pixels classified as MRSA. The HSI was able to successfully discriminate samples in replicas that were sensitive and resistant, based on the calibration model it received.

Characterization of T Helper 1 and 2 Cytokine Profiles in Newborns of Mothers with COVID-19.

An infectious disease caused by SARS-CoV-2, COVID-19 greatly affects the pediatric population and is 3 times more prevalent in newborns than in the general population. In newborns, the overexpression of immunological molecules may also induce a so-called cytokine storm. In our study, we evaluated the expression of cytokines in newborns admitted to a neonatal ICU whose mothers had SARS-CoV-2 and symptoms of SARS. The blood of newborns of infected and healthy mothers was collected to identify their Th1 and Th2 cytokine profiles, and via flow cytometry, the cytokines TNF-α, IFN-γ, IL-2, IL-6, and IL-10 were identified. Overexpression was observed in the Th1 and Th2 cytokine profiles of newborns from infected mothers compared with the control group. Statistical analysis also revealed significant differences between the cellular and humoral responses of the infected group versus the control group. The cellular versus humoral responses of the newborns of infected mothers were also compared, which revealed the prevalence of the cellular immune response. These data demonstrate that some cytokines identified relate to more severe symptoms and even some comorbidities. IL-6, TNF-α, and IL-10 may especially be related to cytokine storms in neonates of mothers with COVID-19.

Severe Neurological Manifestation in a Child with Multisystem Inflammatory Syndrome.

BACKGROUND AND OBJECTIVES: During the COVID-19 pandemic, we followed with concern the evolution of several children diagnosed with Multisystem Inflammatory Syndrome in Children (MIS-C). The purpose of this study is to describe the evolution of MIS-C in a previously healthy 3-year-old girl. METHODS: We tracked the daily medical report of all children admitted with suspected MIS-C to the five largest regional hospitals. RESULTS: Our screening identified a child who had several neurological complications associated with MIS-C. We report hematological alterations, transient cardiac dysfunction, and cerebral involvements such as laminar cortical necrosis caused by ischemic stroke. We present the course of treatment and clinical outcome, and other complications such as a severe subglottic stenosis occurring after extubation. CONCLUSION: Subglottic stenosis is an expected complication after prolonged intubation, and the presence of dysphonia and/or stridor is an important predictive factor. MIS-C with severe neurological alteration may occur in a healthy child, and early diagnosis and treatment with a pulse of corticoid with immunoglobulin are essential for a favorable outcome.

Influenza and other respiratory viruses in children: prevalence and clinical features.

With the COVID-19 pandemic still ongoing, the annual season of influenza and other respiratory virus epidemics has arrived. Specimens from patients suspected of respiratory viruses infection were collected. Viral detection was performed following RNA extraction and real-time RT-PCR. During the study period, we received and tested a total of 606 specimens. Rhinovirus virus was the viral type most prevalent, detected in 186 (45.47%) specimens. The age range of patients positive for influenza A, influenza A (H1N1), and influenza B was 18 days to 13 years. With female prevalence for this viral type, cough and asthma were the main clinical manifestations presented by this viral type. Our results indicate that rhinoviruses, adenoviruses, metapneumoviruses, and influenza are among the most important agents of ARI in pediatrics. The epidemic period of respiratory infections observed in Goiânia can be useful for planning and implementing some prevention strategies.

Draft Genome Sequences of Chromobacterium Strains Isolated from Water Systems in Central Western Brazil.

We report the draft genome sequences of four Chromobacterium strains. This report includes the draft genome sequences of four environmental strains, isolated from surface waters in Brazil.

Elevated Mortality Risk from CRKp Associated with Comorbidities: Systematic Review and Meta-Analysis.

Carbapenem-resistant Klebsiella pneumoniae has become a public health problem with therapeutic limitations and high mortality associated with comorbidities. Methods: This is a systematic review and meta-analysis with a search in PubMed, SciELO, and Lilacs. Ten articles were selected, considering cohort, case-control, and cross-sectional studies. Tests for proportions and relative risk of mortality were performed, considering a 5% threshold for significance. Statistical analyses were performe dusing Rstudio® software, version 4.0.2 of Ross Ihaka and Robert Genleman in Auckland, New Zealand. Results: Klebsiella pneumoniae, associated with chronic kidney disease, was responsible for 26%/258 deaths, chronic lung disease 28%/169, diabetes 31%/185, liver disease 15%/262, and heart disease 51%/262 deaths. Carbapenem-resistant Klebsiella pneumoniae associated with chronic kidney disease was responsible for 49%/83 deaths, with diabetes 29%/73, and with liver disease 33%/73 deaths. The risk of death from carbapenem-resistant Klebsiella pneumoniae was twice as high as the number of deaths associated with carbapenem-sensitive Klebsiella pneumoniae, RR = 2.07 (p < 0.00001). Conclusions: The present study showed an increase in mortality from carbapenem-resistant Klebsiella pneumoniae when associated with comorbidities.

Escherichia coli, Species C Human Adenovirus, and Enterovirus in Water Samples Consumed in Rural Areas of Goiás, Brazil.

Rural environments lack basic sanitation services. Facilities for obtaining water and disposing sewage are often under the initiative of each resident, who may not be able to build and maintain them properly. Thus, water for human consumption is subject to fecal contamination and, consequently, the presence of waterborne pathogens, such as enteric viruses. This study evaluated fecal contamination of water samples from individual sources used for domestic water supply on small farms in the state of Goiás, Brazil. Samples were collected from 78 houses whose water sources were tubular wells, dug wells, springs, and surface waters. Escherichia coli (EC) bacteria, analyzed by the defined chromogenic substrate method, was used as a traditional indicator of fecal contamination. The enteric viruses Human mastadenovirus (HAdV) and Enterovirus (EV), analyzed by qPCR, were tested as complementary indicators of fecal contamination. At least one of these markers was found in 89.7% of the samples. Detection rates were 79.5% for EC, 52.6% for HAdV, and 5.1% for EV. The average concentration for EC was 8.82 × 101 most probable number (MPN) per 100 mL, while for HAdV and EV the concentrations were 7.51 × 105 and 1.89 × 106 genomic copies (GC) per liter, respectively. EC was the most frequent marker in ground and surface water samples. HAdV was detected significantly more frequently in groundwater than in surface water and was more efficient in indicating contamination in tubular wells. There was no association of frequencies or correlation of concentrations between EC and HAdV. HAdV indicated human fecal contamination and performed well as a complementary indicator. The results reveal that a large part of the analyzed population is vulnerable to waterborne diseases caused by enteric pathogens.

Spending Estimates for Gastric Cancer in Central Brazil

Abstract Stomach cancer is the second leading cause of death by cancer worldwide and is even more pronounced in South America. In Brazil, it is estimated that an increase in the number of cases due to this cancer occurred in the biennium 2018-2019. In this study, we investigated the expenditures of the State Health Department of Goiás on hospitalizations and treatment of gastric cancer for the Unified Health System (SUS) from 2008-2016. This is a cross-sectional, descriptive, and analytical study based on secondary data from the Unified Health System computing department (DATASUS) and the System of Management of the Table of Procedures, Medications, Orthosis, Prosthesis, and Special Materials of SUS through CONECTA-SUS related to International Classification of Diseases-10/C16 (ICD-10/C16) procedures for gastric neoplasms. A total of I$ 5,697,958.20 was spent on gastric tumor in the last nine years in Goiás, I$ 4,492,916.67 (0.3%) on hospitalizations, and I$ 1,997,120.91 on treatment. This study presents a current and relevant estimate of the costs of gastric cancer patients in Goiás. Moreover, we provide information on the extent of the cancer issue to public health. Our analysis offers components for service management and studies that reduce resource allocation in more rational ways

HELICOBACTER PYLORI cagA VIRULENCE GENE AND SEVERE ESOGASTRODUODENAL DISEASES: IS THERE AN ASSOCIATION?

BACKGROUND: Helicobacter pylori colonizes approximately half of the world's human population. Its presence in the gastric mucosa is associated with an increased risk of gastric adenocarcinoma, gastric lymphoma, and peptic ulcer disease. In Brazil, the high prevalence of H. pylori infection is a serious health problem. H. pylori virulence factors are associated with an increased risk of serious gastrointestinal disorders. The cagA gene encodes a cytotoxin-A-associated antigen (CagA) that is involved in bacterial pathogenicity. H. pylori strains carrying the cag pathogenicity island (cag-PAI) are significantly associated with severe clinical outcomes and histopathological changes. OBJECTIVE: The present study aims to investigate the prevalence of the cagA gene among H. pylori isolates from patients with different gastric pathologies. Further, the study hopes to verify its association with clinical outcomes. In addition, phylogenetic analysis was performed on cagA-positive H. pylori strains from patients with severe and non-severe diseases. METHODS: Gastric specimens were collected through a biopsy from 117 patients with different esogastroduodenal diseases. DNA was extracted from these gastric specimens and the polymerase chain reaction was performed to amplify the gene fragments corresponding to the 16S ribosomal RNA and cagA genes using specific primers. The polymerase chain reaction products of selected samples positive for cagA were sequenced. The sequences were aligned with reference sequences from the National Center for Biotechnology Information (NCBI) (Bethesda/USA), and a phylogenetic tree was constructed. RESULTS: H. pylori was detected in 65.9% (77/117) of Brazilian patients with different gastroduodenal disorders. Overall, 80.5% (62/77) of the strains were cagA-positive. The ages of patients with cagA-positive strains (15 males and 47 females) ranged from 18 to 74 years. The lesions were categorized as non-severe and severe according to the endoscopic and histopathological reports the most prevalent non-severe esogastroduodenal lesion was gastritis 54/77 (70.12%), followed by esophagitis 12/77 (15.58%) and duodenitis 12/77 (15.58%). In contrast, the most prevalent severe lesions were atrophy 7/77 (9.09%), followed by metaplasia 3/77 (3.86%) and gastric adenocarcinoma 2/77 (2.59%). Phylogenetic analyses performed with the partial sequences of the cagA gene obtained from local strains were grouped in the same clade. No differences in phylogenetic distribution was detected between severe and non-severe diseases. CONCLUSION: The cagA gene is highly prevalent among H. pylori isolates from gastric lesions in Brazilian patients. The presence of the cagA gene was not considered a marker of the severity of esogastroduodenal lesions in the present study. This is the first study to investigate the phylogenetic population structure of H. pylori strains in a Brazilian capital, which may improve our understanding of the clinical outcome of H. pylori infection.

HELICOBACTER PYLORI cagA VIRULENCE GENE AND SEVERE ESOGASTRODUODENAL DISEASES: IS THERE AN ASSOCIATION? / Gene de virulência cagA de Helicobacter pylori e doenças esogastroduodenais severas: existe uma associação?

ABSTRACT BACKGROUND: Helicobacter pylori colonizes approximately half of the world's human population. Its presence in the gastric mucosa is associated with an increased risk of gastric adenocarcinoma, gastric lymphoma, and peptic ulcer disease. In Brazil, the high prevalence of H. pylori infection is a serious health problem. H. pylori virulence factors are associated with an increased risk of serious gastrointestinal disorders. The cagA gene encodes a cytotoxin-A-associated antigen (CagA) that is involved in bacterial pathogenicity. H. pylori strains carrying the cag pathogenicity island (cag-PAI) are significantly associated with severe clinical outcomes and histopathological changes. OBJECTIVE: The present study aims to investigate the prevalence of the cagA gene among H. pylori isolates from patients with different gastric pathologies. Further, the study hopes to verify its association with clinical outcomes. In addition, phylogenetic analysis was performed on cagA-positive H. pylori strains from patients with severe and non-severe diseases. METHODS: Gastric specimens were collected through a biopsy from 117 patients with different esogastroduodenal diseases. DNA was extracted from these gastric specimens and the polymerase chain reaction was performed to amplify the gene fragments corresponding to the 16S ribosomal RNA and cagA genes using specific primers. The polymerase chain reaction products of selected samples positive for cagA were sequenced. The sequences were aligned with reference sequences from the National Center for Biotechnology Information (NCBI) (Bethesda/USA), and a phylogenetic tree was constructed. RESULTS: H. pylori was detected in 65.9% (77/117) of Brazilian patients with different gastroduodenal disorders. Overall, 80.5% (62/77) of the strains were cagA-positive. The ages of patients with cagA-positive strains (15 males and 47 females) ranged from 18 to 74 years. The lesions were categorized as non-severe and severe according to the endoscopic and histopathological reports the most prevalent non-severe esogastroduodenal lesion was gastritis 54/77 (70.12%), followed by esophagitis 12/77 (15.58%) and duodenitis 12/77 (15.58%). In contrast, the most prevalent severe lesions were atrophy 7/77 (9.09%), followed by metaplasia 3/77 (3.86%) and gastric adenocarcinoma 2/77 (2.59%). Phylogenetic analyses performed with the partial sequences of the cagA gene obtained from local strains were grouped in the same clade. No differences in phylogenetic distribution was detected between severe and non-severe diseases. CONCLUSION: The cagA gene is highly prevalent among H. pylori isolates from gastric lesions in Brazilian patients. The presence of the cagA gene was not considered a marker of the severity of esogastroduodenal lesions in the present study. This is the first study to investigate the phylogenetic population structure of H. pylori strains in a Brazilian capital, which may improve our understanding of the clinical outcome of H. pylori infection.

RESUMO CONTEXTO: Helicobacter pylori coloniza aproximadamente metade da população humana mundial. A presença do microrganismo na mucosa gástrica está associada a um risco aumentado de adenocarcinoma gástrico, linfoma gástrico e úlcera péptica. No Brasil, a alta prevalência de infecção por H. pylori é um grave problema de saúde. Os fatores de virulência de H. pylori estão associados a risco aumentado de distúrbios gastrointestinais severos. O gene cagA codifica um antígeno associado à citotoxina A (CagA) que está envolvido na patogenicidade bacteriana. As cepas de H. pylori portadoras da ilha de patogenicidade cag (cag-PAI) estão significativamente associadas a desfechos clínicos severos e alterações histopatológicas. OBJETIVO: O presente estudo tem como objetivo investigar a prevalência do gene cagA entre isolados de H. pylori de pacientes com diferentes desordens gástricas, bem como verificar sua associação com desfechos clínicos. Além disso, a análise filogenética foi realizada em cepas de H. pylori cagA-positivas de pacientes com doenças severas e não severas. MÉTODOS: Amostras gástricas foram coletadas por meio de biópsia gástrica de 117 pacientes com diferentes doenças esogastroduodenais. O DNA foi extraído das amostras e utilizado para amplificar os fragmentos gênicos correspondentes aos genes RNA ribossomal 16S e cagA, através da reação em cadeia da polimerase. Os produtos da reação em cadeia da polimerase de amostras selecionadas positivas para cagA foram sequenciados e as sequências foram alinhadas com sequências de referência do National Center for Biotechnology Information (NCBI) (Bethesda/EUA). As análises filogenéticas foram realizadas a partir do sequenciamento e construção da árvore filogenética. RESULTADOS: H. pylori foi detectado em 65,9% (77/117) dos pacientes brasileiros com diferentes distúrbios gastroduodenais. No total, 80,5% (62/77) das cepas foram cagA-positivas. As idades dos pacientes com cepas cagA-positivas (15 homens e 47 mulheres) variaram de 18 a 74 anos. As lesões foram categorizadas como não severas e severas de acordo com o laudo endoscópico e histopatológico. A lesão esogastroduodenal não severa mais prevalente foi gastrite 54/77 (70,12%), seguida de esofagite 12/77 (15,58%) e duodenite 12/77 (15,58%). Em contraste, as lesões severas mais prevalentes foram atrofia 7/77 (9,09%), seguida de metaplasia 3/77 (3,86%) e adenocarcinoma gástrico 2/77 (2,59%). As análises filogenéticas realizadas com as sequências parciais do gene cagA obtidas de cepas locais foram agrupadas no mesmo clado. Nenhuma diferença na distribuição filogenética foi detectada entre doenças severas e não severas. CONCLUSÃO: O gene cagA é altamente prevalente entre isolados de H. pylori de lesões gástricas em pacientes brasileiros. A presença do gene cagA não foi considerada um marcador de severidade das lesões esogastroduodenais no presente estudo. Este é o primeiro estudo a investigar a estrutura filogenética da população de cepas de H. pylori em uma capital brasileira. Esses resultados irão contribuir para o entendimento sobre o desfecho clínico da infecção por H. pylori.

Risk to human health from protozoan (oo)cysts in water treatment plants / Risco à saúde humana por (oo)cistos de protozoários em estações de tratamento de água

ABSTRACT The presence of waterborne pathogens, when associated with the water supply system, poses risks to public health. This study investigated the occurrence of (oo)cysts of Cryptosporidium spp. and Giardia spp. in two Water Treatment Plants, with full-cycle technology, and assessed the microbiological risk to consumers' health. The membrane filtration technique was employed to identify the protozoan load in 24 samples of raw and filtered water. Physicochemical and bacteriological parameters were also analyzed. The Quantitative Microbial Risk Assessment model was used to determine the risk of daily and annual infection associated with the ingestion of filtered water. A total of 66.67% of raw water samples were positive for Cryptosporidium; and 33.33%, for Giardia. The maximum concentrations of 200 (oo)cysts/L and 50 cysts/L were detected in the Cerrado stream (located in Sanclerlândia, state of Goiás, Brazil), due to the predominance of grazing areas and intense agricultural activity. Water Treatment Plants did not completely remove the protozoa and the retention efficiency was lower than that recommended by the United States Environmental Protection Agency, with average values of 1.27 log for cysts and 1.04 log for oocysts. The probability of annual infection by cysts (100%) was higher than that of oocysts (86.61 - 98.32%) as for consumption of filtered water, and in the dry season, there was a higher risk of infection, due to the low performance of the Water Treatment Plants and higher concentration of pathogens. According to the results, the continuous intake of filtered water above the warning level can cause infectious diseases in the supplied population.

RESUMO A presença de patógenos de veiculação hídrica quando associados ao sistema de abastecimento de água causam risco à saúde pública. Esse estudo investigou a ocorrência de (oo)cistos de Cryptosporidium spp. e Giardia spp. em duas Estações de Tratamento de Água com tecnologia do tipo ciclo completo e avaliou o risco microbiológico à saúde dos consumidores. Utilizou-se a técnica de filtração em membranas para identificar a carga dos protozoários nas 24 amostras de água bruta e filtrada. Os parâmetros físico-químicos e bacteriológicos também foram analisados. O modelo de Avaliação Quantitativa de Risco Microbiológico foi empregado para determinar o risco de infecção diária e anual associada à ingestão de água filtrada. Um total de 66.67% das amostras de água bruta foram positivas para Cryptosporidium; e 33.33% para Giardia. As concentrações máximas de 200 oocistos/L e 50 cistos/L foram detectadas no ribeirão Cerrado (localizado em Sanclerlândia, Estado de Goiás, Brasil), devido ao predomínio de área de pastagens e à intensa atividade agropecuária. As Estações de Tratamento de Água não removeram completamente os protozoários e a eficiência de retenção foi inferior ao recomendado pela Agência de Proteção Ambiental dos Estados Unidos, com valores médios de 1,27 log para cistos e 1,04 log para oocistos. A probabilidade de infecção anual por cistos (100%) foi superior aos oocistos (86,61% a 98,32%) para o consumo da água filtrada, sendo que a estação seca, o risco de infecção foi maior, devido ao baixo desempenho das Estações de Tratamento de Água e à maior concentração de patógenos. Esses resultados indicam que a ingestão contínua de água filtrada acima do nível de alerta pode provocar doenças infecciosas na população abastecida.

Helicobacter pylori virulence dupA gene: risk factor or protective factor?

Helicobacter pylori is the etiological agent of chronic gastritis, peptic ulcer, and gastric cancer. The duodenal ulcer-promoting gene dupA, which is located in the plasticity region of the H. pylori genome, is homologous to the virB gene which encodes a type IV secretion protein in Agrobacterium tumefaciens. Studies have shown associations between H. pylori dupA-positive strains and gastroduodenal diseases. However, whether dupA acts as a risk factor or protective factor in these diseases remains unclear. Therefore, in this study, we aimed to verify the presence of the dupA gene in infectious H. pylori strains in the Brazilian mid-west and to investigate its association with the clinical outcomes of patients with dyspepsia. Additionally, the phylogenetic origin of the strains was determined. Gastric biopsies from 117 patients with dyspepsia were analyzed using histological and molecular techniques. The hpx gene (16S rRNA) was used to screen for H. pylori infection, and positive samples were then subjected to dupA gene detection and sequencing. The estimated prevalence of H. pylori infection was 64.1%, with the dupA gene being detected in a high proportion of infectious strains (70.7%). Furthermore, a risk analysis revealed that for women, a dupA-positive H. pylori infection increased the chance of developing gastritis by twofold. The partial dupA sequences from isolated infectious strains in this work are similar to those of strains isolated in westerns countries. This study provides useful insights for understanding the role of the H. pylori dupA gene in disease development.

Physicochemical, microbiological quality, and risk assessment of water consumed by a quilombola community in midwestern Brazil.

The quality of the water consumed by a given community is related to its quality of life. In this sense, this study aimed to evaluate, from the perspective of health risk, the physical, chemical, and microbiological quality of drinking water, in a quilombola community, and the qualitative aspects intrinsic to its use and storage. For this, water samples, collected at the exits of the collective water supply system and from eight cisterns that store rainwater, used for human consumption, were analyzed. The samples were subjected to physical, chemical, and microbiological analysis, including adenovirus (HAdV) and enterovirus (EV). The probability of an individual acquiring infection through water consumption was determined by quantitative microbiological risk analysis using HAdV and Escherichia coli (EC) as reference pathogens. The results showed that the water in the deep tubular well had 270.8 mg/L of total hardness, leading to the rejection of its consumption by ingestion. Alternativity, the people in the community consume rainwater stored in cisterns. For this type of water, the presence of heterotrophic bacteria was found in 75%, total coliform was present in 100%, and Enterococci were detected in 25%. Furthermore, EC was present in 25%, EV in 50%, and HAdV in 100% of the samples. The probability of annual infection with HAdV and EC was, in the worst situation, 100% and 1.3%, respectively. Regarding the qualitative and quantitative aspects, there was a significant positive correlation between the absence of EC and the withdrawal of water from the cistern using a pump and the opposite when the withdrawal was carried out using a bucket or hose. Based on the results found, it is important to carry out actions aimed at improving water quality and, consequently, the quality of life of people living in the study community.

Decrease of miR-125a-5p in Gastritis and Gastric Cancer and Its Possible Association with H. pylori.

PURPOSE: The aim of this study was to evaluate the expression of miR-125a-5p in patients with dyspeptic symptoms and gastric cancer, correlating them with the development of this cancer and H. pylori. METHODS: Patients were divided in groups according to histopathological analysis (control, gastritis, and cancer groups). Polymerase chain reaction was performed to detect H. pylori and real-time quantitative PCR to determine miR-125a-5p expression. RESULTS: H. pylori was detected in 44% of the patients, with prevalence in the gastritis and cancer groups. A statistically significant decrease of miR-125a-5p expression was found in the control positive (p = 0.0183*), gastritis positive (p = 0.0380*), and cancer positive (p = 0.0288*) groups when compared with the control negative group. CONCLUSION: We suggest that decreased expression of the miRNA-125a-5p associated with the presence of the H. pylori is an important mechanism in gastric diseases and could be a possible marker for early diagnosis of gastric cancer.

Detection of resistance genes in pyometra isolated bacteria in bitches / Detecção de genes de resistência em bactérias isoladas de piometra em cadelas

Pyometra has several immunological and molecular changes that are responsible for uterine inflammation and the disease may or may not have infections. This study aimed to isolate and identify bacteria in the uterine content of bitches with pyometra, to analyze the susceptibility profile to antibiotics, detect ß-lactamase enzyme production by phenotypic tests, and resistance genes to ß-lactams. Eighteen samples of uterine content were collected by aspiration puncture. The samples were inoculated in bacteriological media and identified by biochemical tests. Subsequently, antibiogram tests, screening for detection of ß-lactamases, and Real-Time PCR for detection of resistance genes was performed. Escherichia coli, Klebsiella spp., Enterobacter aerogenes, Citrobacter spp., Staphylococcus spp., and Streptococcus spp. were identified in the analyzed samples of uterine content. In the antibiogram test, 90.5% of the isolates showed resistance to at least one antibiotic, and of these, 36.8% were considered MDR, with three Staphylococcus spp., three E. coli, and one Klebsiellaspp. Concerning bacterial resistance to the groups of antibiotics tested, 38.1% of the isolates were resistant to at least one type of ß-lactam, 33.3% to tetracycline, 19.0% to aminoglycosides, and 14.3% to fluoroquinolones, macrolides, and trimethoprim-sulfamethoxazole. In the phenotypic test to detect ß-lactamase production, E. coli samples were negative and Klebsiella spp. was positive for the production of AmpC, which presented the blaCMY, blaSPM, and blaSIM genes. Bacteria that are resistant to antibiotics represent a great challenge and laboratory support is therefore essential, without which therapeutic success decreases and death may be inevitable.(AU)

A piometra apresenta diversas alterações imunológicas e moleculares que são responsáveis pela inflamação uterina, e a doença pode ser infecciosa ou não. O objetivo deste estudo foi isolar e identificar bactérias no conteúdo uterino de cadelas com piometra, analisar o perfil de suscetibilidade aos antibióticos, detectar a produção de enzimas ß-lactamase por testes fenotípicos e genes de resistência aos ß-lactâmicos. Dezoito amostras de conteúdo uterino foram coletadas por punção aspirativa. As amostras foram inoculadas em meio bacteriológico e identificadas por testes bioquímicos. Posteriormente, foram realizados testes de antibiograma, triagem para detecção de ß-lactamases e PCR em tempo real para detecção de genes de resistência. Escherichia coli, Klebsiella spp., Enterobacter aerogenes, Citrobacter spp., Staphylococcus spp. e Streptococcus spp. foram identificados nas amostras de conteúdo uterino analisadas. No teste de antibiograma, 90,5% dos isolados apresentaram resistência a pelo menos um antibiótico, e destes, 36,8% foram considerados MR, sendo três Staphylococcus spp., três E. coli e uma Klebsiella spp. Sobre a resistência bacteriana aos grupos de antibióticos testados, 38,1% dos isolados foram resistentes a pelo menos um tipo de ß-lactâmico, 33,3% à tetraciclina, 19,0% aos aminoglicosídeos e 14,3% às fluorquinolonas, macrolídeos e trimetoprim-sulfametoxazol. No teste fenotípico para detecção da produção de ß-lactamase, as amostras de E. coli foram negativas, e Klebsiella spp. foi positiva para a produção de AmpC, que apresentou os genes blaCMY, blaSPM e blaSIM. As bactérias resistentes aos antibióticos representam um grande desafio e, portanto, o suporte laboratorial é essencial, sem o qual o sucesso terapêutico diminui e a morte pode ser inevitável.(AU)

Diagnostic performance between histopathological and molecular methods in the detection of Helicobacter pylori / Desempenho diagnóstico entre os métodos istopatológico e molecular na detecção de Helicobacter pylori

ABSTRACT Introduction: Helicobacter pylori (H. pylori) is a Gram negative bacterium considered to be the etiologic agent of various gastric diseases. The prevalence of bacterial infection varies according to age, geographic location, ethnicity and socioeconomic status. The chronic infection caused by microorganism can favor the development of severe pathologies such as gastric adenocarcinoma. In this sense, early diagnosis is essential for a better prognosis and therapeutic success. Several diagnostic methods performed using invasive and non-invasive techniques, with different sensitivity and specificity, have been used in the detection of H. pylori. Objective: To compare the performance of the molecular and histopathological technique used in the diagnosis of H. pylori infection. Methods: 76 gastric tissue samples were collected from dyspeptic patients who underwent molecular and histopathological diagnosis. Molecular detection was performed using the ribosomal gene (16S rRNA) using the polymerase chain reaction (PCR) technique. Results: The PCR-based molecular diagnostic method detected the bacterium in 63.1% of the samples, while the histopathological test identified the microorganism in only 38.1% of gastric biopsies. The data demonstrated that the PCR technique was about 1.6 times more sensitive than the histopathological technique. Conclusion: The PCR technique was the most efficient diagnostic method for detecting H. pylori and can be implemented in the laboratory routine as a complementary test for the early detection of H. pylori.

RESUMEN Introducción: Helicobacter pylori (H. pylori) es una bacteria Gram negativa considerada agente etiológico de diversas enfermedades gástricas. La prevalencia de la infección bacteriana varía según la edad, la ubicación geográfica, la etnia y el nivel socioeconómico. La infección crónica provocada por esos microorganismos puede favorecer el desarrollo de patologías graves como el adenocarcinoma gástrico. Por esa razón, el diagnóstico precoz es fundamental para un mejor pronóstico y éxito terapéutico. En la detección de H. pylori se han utilizado varios métodos de diagnóstico realizados mediante técnicas invasivas y no invasivas, con diferentes sensibilidad y especificidad. Objetivo: Comparar el desempeño de las técnicas moleculares e histopatológicas utilizadas en el diagnóstico de la infección por H. pylori. Métodos: Se recolectaron 76 muestras de tejido gástrico de pacientes dispépticos y se las sometieron a diagnóstico molecular e histopatológico. La detección molecular se realizó mediante el gen ribosómico (ARNr 16S) mediante la técnica de reacción en cadena de la polimerasa (PCR). Resultados: El método molecular de diagnóstico basado en PCR detectó la bacteria en el 63,1% de las muestras, mientras que la prueba histopatológica identificó el microorganismo en solo el 38,1% de las biopsias gástricas. Los datos demostraron que la técnica de PCR era aproximadamente 1,6 veces más sensible que la técnica histopatológica. Conclusión: La técnica de PCR fue el método diagnóstico más eficaz para la detección de H. pylori y puede implementarse en la rutina del laboratorio como prueba complementaria para la detección precoz de H. pylori.

RESUMO Introdução: Helicobacter pylori (H. pylori) é uma bactéria Gram negativa considerada o agente etiológico de várias doenças gástricas. A prevalência da infecção bacteriana varia de acordo com idade, localização geográfica, etnia e status socioeconômico. A infecção crônica ocasionada por esse microrganismo pode favorecer o desenvolvimento de patologias severas, como o adenocarcinoma gástrico. Nesse sentido, o diagnóstico precoce é essencial para um melhor prognóstico e o sucesso terapêutico. Vários métodos de diagnóstico realizados com técnicas invasivas e não invasivas, com diferentes sensibilidade e especificidade, têm sido utilizados na detecção de H. pylori. Objetivo: Comparar o desempenho das técnicas molecular e histopatológica utilizadas no diagnóstico da infecção por H. pylori. Métodos: Setenta e seis amostras de tecido gástrico foram coletadas de pacientes dispépticos e submetidas ao diagnóstico molecular e histopatológico. A detecção molecular foi realizada utilizando o gene ribossomal (rRNA 16S) por meio da técnica de reação em cadeia da polimerase (PCR). Resultados: O método molecular de diagnóstico com base na PCR detectou a bactéria em 63,1% das amostras, enquanto o teste histopatológico identificou o microrganismo em apenas 38,1% das biópsias gástricas. Os dados demonstram que a técnica de PCR apresentou cerca de 1,6 vezes mais sensibilidade que a técnica histopatológica. Conclusão: A técnica de PCR foi o método de diagnóstico mais eficiente para detecção de H. pylori e pode ser implementada na rotina laboratorial como teste complementar para detecção precoce de H. pylori.