Metataxonomic Studies to Evaluate the Beneficial Effect of Enzymatic Pretreatment on the Anaerobic Digestion of Waste Generated in Turkey Farming.

Turkey litter waste is lignocellulosic and keratinous, requiring prior enzymatic treatment to facilitate fiber hydrolysis and utilization by microorganisms in anaerobic digestion (AD) process. The understanding of the performance of microorganisms in AD can be facilitated through molecular biology and bioinformatics tools. This study aimed to determine the taxonomic profile and functional prediction of microbial communities in the AD of turkey litter waste subjected to enzymatic pretreatment and correlate it with operational parameters. The tests involved the use of turkey litter (T) at 25 g L-1 of volatile solids, a granular inoculum (S) (10% m/v), and the addition of cellulase (C), and pectinase (P) enzymes at four concentrations. The use of enzymes increased methane production by 19% (turkey litter, inoculum, and cellulase-TSC4) and 15% (turkey litter, inoculum, and enzymatic pectinase-TSP4) compared to the control (turkey litter and inoculum-TS), being more effective in TSC4 (667.52 mLCH4), where there was consumption of acetic, butyric, and propionic acids. The pectinase assay (TSP4) showed a methane production of 648 mLCH4 and there was the accumulation of metabolites. Cellulolytic microorganisms Bacteroides, Ruminofilibacter, Lachnospiraceae, Ruminococcaceae, and Methanosaeta were favored in TSC4. In TSP4, the predominant genus was Macellibacteroides and Methanosarcina, and genes involved in methylotrophic methanogenesis were also found (mtaB, mtmB, and mtbB). Enzymes involved in hydrogenotrophic methanogenesis were identified in both assays (TSC4 and TSP4). Molecular tools helped to understand the metabolic routes involved in AD with enzymatic treatment, allowing the elaboration of strategies to improve the sustainable degradation of turkey litter waste.

Metataxonomic characterization of the microbial present in the anaerobic digestion of turkey litter waste with the addition of two inocula: allochthonous and commercial.

Turkey litter waste is lignocellulosic waste that can be sustainably used as an energy source through anaerobic digestion (AD). The 16S ribosomal RNA technique helps to unravel microbial diversity and predominant metabolic pathways. The assays were performed in 600-mL-glass bottles with 400 mL volume, for 60 days at 37 °C. The study evaluated the physicochemical parameters, the composition of the microbiota, and the functional inference in AD of different concentrations of turkey litter (T) using two inocula: granular inoculum (S) and commercial inoculum (B). The highest accumulated methane production (633 mL CH4·L-1) was observed in the test containing 25.5 g VS·L-1 of turkey litter with the addition of the two inocula (T3BS). In tests without inoculum (T3) and with commercial inoculum (T3B), there was an accumulation of acids and consequent inhibition of methane production 239 mL CH4·L-1 and 389 mL CH4·L-1, respectively. Bacteroidota, Firmicutes, and Actinobacteria were the main phyla identified. The presence of archaea Methanobacterium, Methanocorpusculum, and Methanolinea highlighted the hydrogenotrophic metabolic pathway in T3BS. Functional prediction showed enzymes involved in three metabolic pathways in turkey litter biodigestion: acetotrophic, hydrogenotrophic, and methylotrophic methanogenesis. The predominant hydrogenotrophic pathway can be observed by analyzing the microbiota, archaea involved in this specific pathway, genes involved, and relative acid consumption for T3S and T3BS samples with higher methane production. Molecular tools help to understand the main groups of microorganisms and metabolic pathways involved in turkey litter AD, such as the use of different inocula, allowing the development of strategies for the sustainable disposal of turkey litter.

Post-COVID-19 Intensive Care Unit-Acquired Weakness Compromises Long-Term Functional Status.

OBJECTIVE: The objective of this study was to evaluate the impact of intensive care unit (ICU)-acquired weakness (ICUAW) on the functional independence of patients hospitalized for coronavirus disease 2019 (COVID-19) over 6 months after ICU discharge. METHODS: This was a prospective cohort study that included patients who were admitted to the ICU because of COVID-19 and who were monitored for 6 months after discharge from the ICU via telephone. Patients were evaluated at 3 times (30 days, 3 months, and 6 months after discharge from the ICU) for functional independence for personal care and mobility activities (Barthel Scale), independence for self-care (Katz Index), impact of COVID-19 on functional status (post-COVID-19 Functional Status Scale [PCFS]), and mobility level (ICU Mobility Scale). The existence of some degree of dependence was considered when the Barthel Scale score was <100 points, the Katz Index was ≥1, and the PCFS score was ≥1. A PCFS score of ≥3 indicated moderate or severe dependence. Patients with a Medical Research Council score of <48 at discharge from the ICU were diagnosed with ICUAW. RESULTS: Sixty-eight patients were included, with a mean age of 51 (SD = 13) years. The ICUAW rate at ICU discharge was 35%. In the evaluation with the PCFS, the values for the presence of any functional limitation at 30 days, 3 months, and 6 months after ICU discharge were 89.7%, 57.4%, and 38.2%, respectively. The rate of persistence of functional limitations after 6 months was higher in patients with ICUAW than in those without ICUAW (66.7% vs 22.8%; P = .000); the same was true for moderate or severe limitations (20.8% vs 4.5%; P = .035). Likewise, functional independence for personal care, mobility, and self-care activities was poorer in patients with ICUAW. CONCLUSIONS: In patients surviving an ICU stay due to COVID-19, decreased functional independence persists even 6 months after discharge, and patients with ICUAW have worse outcomes. IMPACT: Patients who survive ICU stays due to COVID-19 continue to have greater functional dependence even 6 months after ICU discharge.

Metataxonomic characterization of the microbial community involved in the production of biogas with microcrystalline cellulose in pilot and laboratory scale.

Biogas, produced in anaerobic digestion, is a sustainable alternative for generating energy from agro-industrial and municipal waste. Information from the microbiota active in the process expands the possibilities for technological innovation. In this study, taxonomic annotations, and functional prediction of the microbial community of the inoculum of two processes were carried out: an industrial unit (pilot-scale urban solid waste plant-IU) and a laboratory-scale reactor fed with swine and cattle waste (LS). The biochemical potential of biogas was obtained using tested inoculum with microcrystalline cellulose, obtaining 682 LN/kgVS (LSC-laboratory scale inoculum and microcrystalline cellulose), and 583 LN/kgVS (IUC-industrial unit inoculum and microcrystalline cellulose), which is equivalent to a recovery of 91.5% of total biogas to LSC. The phyla Synergistota and Firmicutes were more abundant in LS/LSC. In the IU/IUC (treatment of restaurant waste and customs seizures), there was a greater microbiological variety and a predominance of the Bacteroidota, Cloacimonadota, Firmicutes and Caldatribacteriota. The genus Methanosaeta predominated in the process, and it was possible to infer the genes (K01895, K00193 and K00625) related to acetoclastic pathway, as well as endoglucanases that are involved in the metabolism of cellulose (LSC). Terpenoids, polyketides, cofactors, and vitamin metabolism were higher in reactors that received different substrates (IU; IUC). The taxonomic and functional differences revealed the importance of determining the microbiota in the analysis of the potential of an inoculum, combined with the use of microcrystalline cellulose, which can provide optimization information in the production of clean energy.

Canine parvovirus type 2 vaccines in Brazil: Viral load in commercial vaccine vials and phylogenetic analysis of the vaccine viruses.

Canine parvovirus type 2 (CPV-2) is the etiological agent of a highly contagious and frequently fatal disease in dogs. Live attenuated vaccines (LAV) are recommended to prevent and control this disease. Commercial vaccines are typically produced with CPV-2 strains adapted to cell culture and usually non-pathogenic. The present study aimed to determine the viral load of CPV-2 vaccines commercially available in Brazil and to characterize the vaccine virus by DNA analysis of its capsid gene. The results demonstrated that all vaccine strains presented high homology of the VP2 gene and they were all closely related to the original CPV-2 strains. However, vaccine strains presented several differences in comparison with field strains currently circulating in Brazil. Seventy-one vials contained viral loads ranging from 7.4E3 to 4.9E10 DNA copies/ml. Nine vials did not contain any detectable CPV-2 DNA. In conclusion, there are genetic and antigenic differences among CPV-2 vaccines and field strains. Additionally, some vaccines have been commercialized with low titers of CPV-2. It is important to improve the quality of the vaccines to prevent or reduce the spread of CPV-2 in Brazil.

Seasonal characterization of the prokaryotic microbiota of full-scale anaerobic UASB reactors treating domestic sewage in southern Brazil.

Upflow Anaerobic Sludge Blanket (UASB) reactors are alternatives in the anaerobic treatment of sanitary sewage in different parts of the world; however, in temperate environments, they are subject to strong seasonal influence. Understanding the dynamics of the microbial community in these systems is essential to propose operational alternatives, improve projects and increase the quality of treated effluents. In this study, for one year, high-performance sequencing, associated with bioinformatics tools for taxonomic annotation and functional prediction was used to characterize the microbial community present in the sludge of biodigesters on full-scale, treating domestic sewage at ambient temperature. Among the most representative phyla stood out Desulfobacterota (20.21-28.64%), Proteobacteria (7.48-24.90%), Bacteroidota (10.05-18.37%), Caldisericota (9.49-17.20%), and Halobacterota (3.23-6.55%). By performing a Canonical Correspondence Analysis (CCA), Methanolinea was correlated to the efficiency in removing Chemical Oxygen Demand (COD), Bacteroidetes_VadinHA17 to the production of volatile fatty acids (VFAs), and CI75cm.2.12 at temperature. On the other hand, Desulfovibrio, Spirochaetaceae_uncultured, Methanosaeta, Lentimicrobiaceae_unclassified, and ADurb.Bin063-1 were relevant in shaping the microbial community in a co-occurrence network. Diversity analyses showed greater richness and evenness for the colder seasons, possibly, due to the lesser influence of dominant taxa. Among the principal metabolic functions associated with the community, the metabolism of proteins and amino acids stood out (7.74-8.00%), and the genes related to the synthesis of VFAs presented higher relative abundance for the autumn and winter. Despite the differences in diversity and taxonomic composition, no significant changes were observed in the efficiency of the biodigesters.

Microbiota profile in mesophilic biodigestion of sugarcane vinasse in batch reactors.

The vinasse is a residue of ethanol production with the potential for methane production, requiring an allochthonous inoculum. Several microorganisms act in the different phases of anaerobic digestion, and the identification of these microbial communities is essential to optimize the process. The characterization of the microbiota involved in the biodigestion of vinasse was observed in the initial stage (IS), at the peak of methane production (MS) and the end of the process (FS) of the best performance assay by high-throughput sequencing. The highest methane production was 0.78 mmolCH4.gVS.h-1 at 243.7 h in the substrate/inoculum ratio of 1.7, with consumption partial of acetic, propionic and isobutyric acids and an 82% reduction of chemical oxygen demand. High microbial diversity was found. The genera Clostridium, Acinetobacter, Candidatus Cloacamonas, Bacteroides, Syntrophomonas, Kosmotoga, the family Porphyromonadaceae and the class Bacteroidia were the most abundant in the maximum methane production. Methane production was driven by Methanobacterium and Methanosaeta, suggesting the metabolic pathways used were hydrogenotrophic and acetoclastic.

Variation of the Prokaryotic and Eukaryotic Communities After Distinct Methods of Thermal Pretreatment of the Inoculum in Hydrogen-Production Reactors from Sugarcane Vinasse.

The aim of this study is to evaluate the effect of different thermal pretreatments of the inoculum on the diversity of the microbial community producing hydrogen from sugarcane vinasse. High-throughput sequencing of the 16S and 18S rRNA genes was performed. The reactor samples were also selected for the isolation of strict anaerobes. Decreased microbial diversity was observed with increasing pretreatment temperatures, with Firmicutes predominating: 90% to 97%. The highest abundance of Staphylococcus (7.9%) was found in pretreatment at 120 °C / 20 min at pH 6. The fungal analysis revealed a high prevalence of Candida (47%), Agaricomycetes, Pezizomycotina and Aspergillus in assays with higher H2 production (90° C / 10 min at pH 6). Three species of Clostridium were isolated: C. bifermentans, C. saccharoperbutylacetonicum and C. saccharobutylicum. The isolates were tested separately and in co-cultures for the production of hydrogen. Hydrogen-producing capacity by co-culture of Clostridium species was increased by 18%. Knowing microorganisms and understanding the interaction between eukaryotes and prokaryotes is essential to obtain strategies for biotransformation of vinasse for the production of bioenergy.

Production of antimicrobial metabolites against pathogenic bacteria and yeasts by Fusarium oxysporum in submerged culture processes.

The antimicrobial activity of the metabolites produced by Fusarium oxysporum PR-33 in submerged culture was evaluated against Gram-positive and Gram-negative bacteria and yeasts. Metabolites were determined by HPLC-DAD-MS/MS. An extract was obtained following the removal of mycelium by centrifugation and lyophilisation of the supernatant. The compounds in this extract demonstrated broad-spectrum antimicrobial action, with rates of inhibition between 60 and 80%, depending on the species and extract tested. The major compounds of the extracts were identified as fusarinolic acid and its isomer [56.9% flask extract (FE)] and 59.2% bioreactor extract (BE), dehydrofusaric acid (35.7% FE and 31.6% BE), and fusaric acid (6.5% FE and 1.1% BE). Fusaric acid has been shown to be responsible for antimicrobial activity. The cytotoxicity of the extracts was evaluated in culture of HEK-293 and SH-SY5Y animal cells and toxicity of these extracts was verified even in the lowest tested concentrations. Therefore, our results indicate that the compounds identified exhibit potential as antimicrobial agents.

Microbial risk assessment in recreational freshwaters from southern Brazil.

In this study, total coliforms (TC), Escherichia coli, enterovirus (EV), rotavirus (RV), and human mastadenovirus species C and F (HAdV-C and HAdV-F) were evaluated in water samples from Belo Stream. For HAdV-C and F, the infectivity was assessed by integrated cell culture quantitative real-time polymerase chain reaction (ICC-qPCR). Samples were collected monthly (May/2015 to April/2016) at four sites. Viral analyses were performed for both ultracentrifuge-concentrated and unconcentrated samples. For site P4 (used for recreational purposes), QMRA was applied to estimate health risks associated with exposure to E. coli and HAdV-C and F. TC and E. coli were present throughout the collection period. EV and RV were not detected. HAdV-C were present in 8.51% (1.89E + 06 to 2.28E + 07 GC (Genomic Copies)/L) and 21.27% (2.36E + 05 to 1.29E + 07 GC/L) for unconcentrated and concentrated samples, respectively. For HAdV-F were 12.76% (2.77E + 07 to 3.31E + 08 GC/L) and 48.93% (1.10E + 05 to 4.50E + 08 GC/L) for unconcentrated and concentrated samples, respectively. For unconcentrated samples, infectivity for HAdV-C was detected in 37.20% (1st ICC-qPCR) and 25.58% (2nd ICC-qPCR). For HAdV-F, infectivity was detected in 6.97% (1st ICC-qPCR) and 6.97% (2nd ICC-qPCR). For concentrated samples, HAdV-C infectious was observed in 17.02% (1st ICC-qPCR) and in 8.51% (2nd ICC-qPCR). For HAdV-F, were present in 8.51% for both 1st and 2nd ICC-qPCR. Statistical analyzes showed significant difference between the collection sites when analyzed the molecular data of HAdV-F, data of TC and E. coli. Correlation tests showed direct correlation between HAdV-F with E. coli and TC. E. coli concentrations translated to the lowest estimates of infection risks (8.58E-05 to 2.17E-03). HAdV-F concentrations were associated with the highest infection risks at 9.99E-01 and for group C, 1.29E-01 to 9.99E-01. These results show that commonly used bacterial indicators for water quality may not infer health risks associated with viruses in recreational freshwaters.

Assessment of diversity of adenovirus DNA polymerase gene in recreational waters facilitated by ultracentrifugal concentration.

Adenoviruses (AdV) are related to respiratory and gastrointestinal diseases in animals and human beings. Their wide genetic diversity in water bodies and their resistance to environmental conditions allow the use of AdV as a reliable marker for detection of fecal contamination. In this work, the diversity of AdV along Belo Stream - in the city of Caxias do Sul, Rio Grande do Sul, Brazil - was evaluated. Samples were compared in both concentrated and unconcentrated forms. The identification of different AdV species was performed by amplifying a partial sequence of the DNA polymerase gene. AdV was detected in 24 out of 55 concentrated samples (43.6%) and the following species were identified: human adenovirus (HAdV) species C (4/55; 7.2%), D (6/55; 10.9%), E (2/55; 3.6%), and F (9/55; 16.3%). AdV related to other mammalian hosts, such as bovine adenovirus (1/55, 1.8%) and murine adenovirus (2/55, 3.6%), have also been identified; 23.6% (13/55) of the unconcentrated samples were positive, and identified as HAdV species C (6/55, 10.9%), D (1/55, 1.8%), and F (6/55, 10.9%). Results obtained evidenced the presence and the great diversity of AdV, mainly of human origin, circulating in Belo Stream. As expected, the concentration step performed helped to detect AdV in more samples.

Histological markers of degeneration and regeneration of the human intervertebral disk / Marcadores histológicos de degeneração e regeneração do disco intervertebral humano / Marcadores histológicos de la degeneración y regeneración del disco intervertebral humano

ABSTRACT Objective: To define histological scores for intervertebral disc degeneration that would enable the definition of morphological characteristics of disease, besides improving knowledge of the lumbar degenerative disc disease by means of immunohistochemical markers. Methods: Hematoxylin and Eosin, Alcian/PAS, Masson Trichrome and Safranin O/FCF staining was used on the intervertebral disc degeneration sections of patients with lumbar degenerative disc disease. The protein markers defined in immunohistochemistry were cell proliferation (Ki-67) and apoptosis (p53). Results: The study data enabled the determination of Safranin O/FCF stain as the most effective one for evaluating parameters such as area, diameter, and number of chondrocyte clusters. The importance of using stains in association, such as Safranin O/FCF, Masson Trichrome, Alcian/PAS and Hematoxylin and Eosin, was also determined, as they are complementary for the histopathological verification of intervertebral disc degeneration. By expressing proteins using the immunohistochemistry technique, it was possible to consider two stages of disc degeneration: cell proliferation with chondrocyte cluster formation, and induction of apoptosis. Conclusion: This study enabled the histological and immunohistochemical characterization to be determined for lumbar degenerative disc disease, and its degrees of evolution, by determining new disc degeneration scores.

RESUMO Objetivo: Definir escores histológicos de degeneração do disco intervertebral que permitam a identificação de características morfológicas da doença, além de melhorar o conhecimento sobre a discopatia degenerativa lombar por meio de marcadores imuno-histoquímicos. Métodos: As colorações histológicas de hematoxilina e eosina, azul de alcian/PAS, tricrômica de Masson e safranina O/FCF foram utilizadas em cortes de disco intervertebral degenerado de pacientes com discopatia degenerativa lombar. Os marcadores proteicos definidos na imuno-histoquímica permitiram a avaliação da proliferação celular (Ki-67) e da apoptose (p53). Resultados: Os dados do estudo permitiram a determinação da coloração de safranina O/FCF como a mais eficaz para avaliar os parâmetros tais como a área, o diâmetro e o número de agrupamentos de condrócitos. Também se determinou a importância do uso das colorações histológicas de forma associada, como safranina O/FCF, tricrômica de Masson, azul de alcian/PAS e hematoxilina e eosina, uma vez que elas são complementares para a verificação histopatológica da degeneração do disco intervertebral. Pela técnica da expressão de proteínas com técnica imuno-histoquímica, foi possível considerar dois estágios de degeneração do disco: proliferação de células com a formação de agrupamentos de condrócitos, seguida pela indução de apoptose. Conclusão: Este estudo permitiu definir a caracterização histológica e imuno-histoquímica em discopatia degenerativa lombar e seus graus de evolução, determinando novos escores de degeneração discal.

RESUMEN Objetivo: Definir valores histológicos de degeneración del disco intervertebral que permitan la definición de las características morfológicas de la enfermedad y mejorar el conocimiento de la enfermedad degenerativa del disco lumbar mediante marcadores inmunohistoquímicos. Métodos: Las coloraciones histológicas con hematoxilina y eosina, azul alcián/PAS, tricrómico de Masson y safranina O/FCF se utilizaron en secciones de los discos intervertebrales degenerados de pacientes con enfermedad degenerativa del disco lumbar. Los marcadores de proteínas definidos por inmunohistoquímica permitieron la evaluación de la proliferación celular (Ki-67), y la apoptosis (p53) . Resultados: Los datos de la determinación permitieron establecer la tinción con safranina O/FCF como la más eficaz para evaluar parámetros tales como el área, diámetro y número de agrupaciones de condrocitos. Se determinó también la importancia del uso asociado de diversas tinciones, como safranina O/FCF, tricrómico de Masson, azul de alcián/PAS y hematoxilina y eosina, ya que son complementarias para la verificación histopatológica de la degeneración del disco intervertebral. Por la técnica de la expresión de proteína con análisis inmunohistoquímica, fue posible establecer dos etapas de la degeneración del disco: la proliferación de células con la formación de agrupaciones de condrocitos y la inducción de la apoptosis. Conclusión: Este estudio permitió definir la caracterización histológica e inmunohistoquímica en la enfermedad degenerativa del disco lumbar y sus grados de evolución, mediante la determinación de nuevas puntuaciones de degeneración del disco.

Incidência de doença diarreica aguda em Caxias do Sul, Rio Grande do Sul, Brasil, em uma série histórica de 10 anos / Incidence of acute diarrheal disease in Caxias do Sul, Rio Grande do Sul, Brazil, in a 10-year historical series

Objetivos: Avaliar a incidência de doença diarreica aguda registrada em Caxias do Sul, em um período de 10 anos, investigando suas correlações com a vacinação contra rotavírus, com a precipitação pluviométrica e com o número de internações hospitalares. Métodos: Foi realizado um estudo descritivo dos registros de doença diarreica aguda, disponibilizados pelo setor de Vigilância Epidemiológica da Secretaria Municipal de Saúde de Caxias do Sul. Resultados: Entre 2004 e 2013 foram registrados 61.246 casos de doença diarreica aguda, sendo que os maiores números ocorreram em bairros de baixa renda. Durante o período de avaliação foram identificados cinco surtos epidêmicos de doença diarreica aguda, em meses de verão e inverno. Não foram encontradas relações entre doença diarreica aguda e precipitação pluviométrica. O número de internações hospitalares por doença diarreica aguda em crianças de zero a quatro anos decresceu a partir de 2006, coincidindo com o início do programa de vacinação contra rotavírus na rede pública, enquanto que os óbitos de todas as idades sofreram flutuações durante o período estudado. Conclusões: A avaliação da incidência de doença diarreica aguda, por longo período, mostra que houve flutuações ao longo do tempo no número de registros e de óbitos em Caxias do Sul. Este estudo revelou também uma redução no número de internações devidas a doença diarreica aguda no período que sucedeu à introdução do programa de vacinação contra rotavírus na rede pública de saúde.

Aims: To evaluate the incidence of acute diarrheal disease (ADD) recorded over a 10-year period in Caxias do Sul, southern Brazil, and its correlations with the vaccination against rotavirus, with rainfall, and with the number of hospitalizations. Methods: A descriptive study was conducted, analyzing the ADD records obtained from the Epidemiological Surveillance sector of the Local Health Department of Caxias do Sul. Results: A total of 61,246 cases of ADD were recorded between 2004 and 2013, most of which occurred in low-income neighborhoods. There were five outbreaks of ADD in the summer and winter months during the study period. No correlations were found between ADD and rainfall. The number of children aged between zero and four years hospitalized for ADD decreased from 2006 onwards, when vaccination against rotavirus was implemented in the public health network whereas the number of deaths across all age groups fluctuated during the study period. Conclusions: The assessment of the long-term incidence of ADD showed a fluctuation in the number of cases and deaths in Caxias do Sul. In addition, hospitalizations for ADD decreased after the implementation of rotavirus vaccination in the public health network.

Detection of rotavirus and norovirus in the elderly population of Caxias doSul, Rio Grande do Sul, Brazil, from 2010 to 2012 / Detection of rotavirus and norovirus in the elderly population of Caxias doSul, Rio Grande do Sul, Brazil, from 2010 to 2012

Aims: Rotavirus (RV) and norovirus (NoV) are the most common cause of diarrhea in children aged less than five years throughout the world. However, RV and NoV have frequently been overlooked as pathogens in elderly individuals. This study examined the frequency of RV and NoV among elderly patients with complaints of diarrhea in the city of Caxias do Sul, southern Brazil.Methods: Between 2010 and 2012, stool samples from patients aged 60 years or older with acute gastroenteritis treated at a private healthcare center were analyzed, seeking to identify RV and NoV. RV detection was performed by latex agglutination (LA) methods and polyacrylamide gel electrophoresis (PAGE). One third of the samples were further tested for NoV by reverse transcription polymerase chain reaction (RT-PCR).Results: A total of 145 stool samples from patients aged 60 to 105 years were analyzed. RV was detected in 6/145 (4.14%) and 5/125 (4.00%) of the samples by LA and PAGE, respectively. NoV was detected in 6/51 (11.76%). Mixed infection (RV and NoV) was detected in a single sample: 1/51 (1.96%).Conclusions: This study adds further evidence that viral agents are involved in the etiology of gastroenteritis in the elderly, contributing significantly to the understanding of RV and NoV infections in the mature population.

Objetivos: Rotavírus (RV) e norovírus (NoV) são a causa mais comum de diarreia em crianças menores de cinco anos em todo o mundo; entretanto, como patógenos em idosos, esses vírus têm sido pouco investigados. Este estudo examinou a frequência de RV e NoV entre idosos com queixas de diarreia na cidade de Caxias do Sul, no sul do Brasil. Métodos: Entre 2010 e 2012 foram analisadas amostras de fezes de pacientes de umCentro de Atenção à Saúde privado, com idade de 60 anos ou mais, com gastroenterite aguda,buscando-se identificar RV e NoV. A detecção de RV foi realizada pelos métodos Aglutinação em Látex (LA) e Eletroforese em Gel de Poliacrilamida (PAGE). Uma terceira parte das amostras foi ainda testada quanto a NoV por Transcrição Reversa e Reação em Cadeia da Polimerase (RTPCR).Resultados: Ao todo foram analisadas 145 amostras de fezes de pacientes com idade entre 60 e 105 anos. RV foi detectado em 6/145 (4,14%) e 5/125 (4,00%) das amostras por LA e PAGE, respectivamente. NoV foi detectado em 6/51 (11,76%). Infecção mista (RV e NoV) foi detectada em uma única amostra: 1/51 (1,96%). Conclusões: Este estudo acrescenta mais evidências de que agentes virais estão envolvidos na etiologia da gastroenterite em idosos, e contribui para a compreensão das infecções por RV e NoV na população idosa.

Human papillomavirus among women with atypical squamous cells of undetermined significance in southern Brazil.

OBJECTIVE: To determine the prevalence of atypical squamous cells of undetermined significance (ASCUS) and human papillomavirus (HPV) genotypes in a population in southern Brazil. METHODS: In a retrospective cross-sectional study, the prevalence of ASCUS was determined among women aged 20-60 years who were referred to a private medical center in Caxias do Sul by a gynecologist for assessment of a cervical condition between January 1, 2010, and September 30, 2011. Histologic and cytologic samples were tested for HPV, and polymerase chain reaction (PCR) was used to genotype any HPV DNA identified. RESULTS: Among the 250 included women, 25 (10.0%) had ASCUS. HPV DNA was found in 15 (60.0%) women with ASCUS and 115 (51.1%) of the 225 without ASCUS. Viral typing showed that 7 (46.7%) HPV-positive women with ASCUS had multiple infections with up to five different genotypes. Both low- and high-risk HPV genotypes were found in ASCUS samples; the most prevalent genotypes were HPV6/HPV11 (affecting 10 [66.7%] women), HPV51 (6 [40.0%]), and HPV16 (6 [40.0%]). CONCLUSION: ASCUS is not an indication of HPV infection. HPV screening and genotyping would benefit women with ASCUS, because treatment can be planned according to risk of carcinogenesis.

Human papillomavirus and infections of the lower genital tractin women with abnormal cervical cytological examination / Papilomavírus humano e infecções do trato genital inferior em mulheres comexame colpocitológico anormal

Aims: To study the associations of positive human papillomavirus (HPV) DNA in the female genital tract with Gardnerella vaginalis, Trichomonas vaginalis and Candida sp. vaginitis, and with possible socio-demographic risk factors for HPV infections.Methods: The study was conducted with 208 patients with abnormal cervical cytological examination assisted at the Gynecology Service of Central Clinic of Caxias do Sul University, Rio Grande do Sul, Brazil, analyzing the presence of HPV DNA by polymerase chain reaction and associating the results to diagnosis of vaginal infections obtained from the medical charts of the patients.Results: HPV DNA was detected in 60.1% of all cases analyzed; 93% of them presented simple infections and 6.9% showed multiple infections. Seventeen viral types were identified, being HPV16 the most frequent (38.8%). At least one of the vaginitis investigated was observed in 57,2% of the patients. Gardnerella vaginalis was found in 45.7%, Candida sp. in 20.2%, and Trichomonas vaginalis in 3.8% of the patients examined. No statistical association between the presence of HPV DNA and other infection of the lower genital tract or socio-demographic risk factors was observed.Conclusions: No association of HPV infection with other infections of the female genital tract or with socio-demographic features could be found. Multiple infections with up to five types of HPV were observed in a low percentage of women. It is difficult to clearly define a group at high risk for HPV infection. All women with abnormal cervical cytological examination are possible carriers of HPV and other infections of the lower genital tract.

Objetivos: Estudar as associações da positividade para o DNA de papilomavírus humano (HPV) no trato genital inferior feminino com as vaginites por Gardnerella vaginalis, Candida sp. e Trichomonas vaginalis e com a presença de possíveis fatores de risco para infecção por HPV.Métodos: O estudo foi realizado com 208 pacientes com exame colpocitológico anormal atendidas no Serviço de Ginecologia do Ambulatório Central da Universidade de Caxias do Sul analisando a presença do DNA de HPV através de reação em cadeia da polimerase e associando os resultados aos diagnósticos de infecção vaginal obtidos nos prontuários das pacientes.Resultados: Foi detectado DNA de HPV em 60,1% de todos os casos analisados , sendo que 93% apresentaram infecções simples e 6,9 % apresentaram infecções múltiplas. Dezessete tipos virais foram identificados, sendo HPV16 o mais frequente (38,8%). Pelo menos uma das vaginites investigadas foi observada em 57,2% dos pacientes. Gardnerella vaginalis foi encontrada em 45,7%, Candida sp. em 20,2%, e Trichomonas vaginalis em 3,8% das pacientes examinadas. Não foi observada nenhuma associação estatística entre a presença do DNA de HPV e outras infecções do trato genital inferior ou fatores sociodemográficos de risco.Conclusões: Não houve associação da infecção pelo HPV com outras infecções do trato genital feminino nem com características sociodemográficas. Múltiplas infecções com até cinco tipos de HPV foram observadas em uma pequena parcela das mulheres. É difícil definir claramente um grupo de risco para a infecção por HPV. Todas as mulheres com exame colpocitológico anormal são possíveis portadoras do HPV e de outras infecções do trato genital inferior.

Matched case-control study evaluating the frequency of the main agents associated with neonatal diarrhea in piglets / Estudo caso-controle pareado avaliando a frequência dos principais agentes causadores de diarréia neonatal em suínos no Rio Grande do Sul

A case-control study was carried out in litters of 1 to 7-day-old piglets to identify the main infectious agents involved with neonatal diarrhea in pigs. Fecal samples (n=276) from piglets were collected on pig farms in the State of Rio Grande do Sul, Brazil, from May to September 2007. Litters with diarrhea were considered cases (n=129) and normal litters (n=147) controls. The samples were examined by latex agglutination test, PAGE, conventional isolating techniques, ELISA, PCR, and microscopic methods in order to detect rotavirus, bacterial pathogens (Escherichia coli, Clostridium perfringens type A and C, and Clostridium difficile), and parasites (Coccidian and Cryptosporidium spp.). Outbreaks of diarrhea were not observed during sampling. At least one agent was detected in fecal samples on 25 out of 28 farms (89.3 percent) and in 16 farms (57.1 percent) more than one agent was found. The main agents diagnosed were Coccidia (42.86 percent) and rotavirus (39.29 percent). The main agents identified in litters with diarrhea were Clostridium difficile (10.6 percent), Clostridium perfringens type A (8.8 percent) and rotavirus (7.5 percent); in control litters, Clostridium difficile (16.6 percent) and Coccidian (8.5 percent). Beta hemolytic Escherichia coli and Clostridium perfringens type C were not detected. When compared with controls, no agent was significantly associated with diarrhea in case litters. These findings stress the need for caution in the interpretation of laboratorial diagnosis of mild diarrhea in neonatal pigs, as the sole detection of an agent does not necessarily indicate that it is the cause of the problem.

Um estudo de caso-controle em leitegadas de um a sete dias de idade foi realizado com o objetivo de identificar os principais agentes infecciosos envolvidos na diarreia neonatal de leitões. As amostras de fezes (n=276) foram coletadas em granjas de suínos no Estado do Rio Grande do Sul, Brasil, no período de maio a setembro de 2007. Leitegadas com diarreia foram consideradas casos (n=147) e leitegadas normais, controles (n=129). As amostras foram examinadas através do teste de aglutinação em látex, PAGE, cultivo, ELISA, PCR e métodos microscópicos para a excreção dos principais agentes de diarreia: virais (rotavirus), bacterianos (Escherichia coli, Clostridium perfringens tipos A e tipo C e Clostridium difficile) e parasitários (coccídeos e Cryptosporidium spp.). Durante o período do estudo não foram observados surtos e a diarréia, quando presente, apresentou-se leve. Pelo menos um agente foi identificado nas amostras fecais de 25 entre 28 granjas (89,3 por cento) analisadas e em 16 granjas (57,1 por cento) mais de um agente foi detectado. Os principais agentes encontrados nas granjas foram coccídeos (42,86 por cento) e rotavírus (39,29 por cento). Os principais agentes detectados nas leitegadas com diarreia foram Clostridium difficile (10,6 por cento), Clostridium perfringens tipo A (8,8 por cento) e rotavírus (7,5 por cento). Por outro lado, nas leitegadas controle os agentes mais prevalentes foram Clostridium difficile (16,6 por cento) e coccídeos (8,5 por cento). E. coli Beta hemolítica e Clostridium perfringens tipo C não foram detectados. O presente estudo de caso-controle demonstrou que nenhum agente infeccioso esteve associado significativamente com diarreia (p>0.05). Esses achados reforçam a necessidade de que haja cuidado na interpretação de resultados de exames laboratoriais em materiais coletados de leitões com diarreia neonatal leve, pois a detecção isolada de um agente infeccioso não indica necessariamente que o mesmo seja a causa do problema.

High prevalence of human papillomavirus type 58 in patients with cervical pre-malignant lesions in southern Brazil.

Cervical cancer is the second most common type of cancer in women worldwide. Several human papillomavirus (HPV) genotypes, sexual behavior, and socioeconomic profile represent major risk factors for the development of this carcinoma. Cervical invasive cancer is preceded by cellular abnormalities that can be identified by cytological or histological exams. In order to determine the prevalence and genotypes of HPV in women with abnormal cytology or histopathology, cervical cell samples from 256 patients were evaluated for the presence of HPV/DNA by polymerase chain reaction (PCR), followed by virus genotyping by restriction fragment length polymorphism (RFLP). A total of 113 samples (51.2%) were HPV/DNA positive. Viral genotyping showed that the most prevalent genotypes were HPV 16 (34.7%) and 58 (13.8%), followed by HPV 33 (9.72%), 11 (8.33%), 18 (5.55%), 53 (5.55%), and 6 (4.2%). Four samples (5.55%) exhibited multiple infections due to the great similarity of socioeconomic characteristics and sexual behavior of HPV positive women, it was not possible to establish a risk profile for female HPV infection.

Infection with human papillomaviruses of sexual partners of women having cervical intraepithelial neoplasia.

Epidemiological studies show that human papillomaviruses (HPV) are strongly related to cervical cancer and cervical intraepithelial neoplasias (CIN). Unlike the case for women, there are no consistent data on the natural history of HPV in the male population even though these viruses are prevalent in males. We carried out a prospective study to assess the prevalence of HPV in males as well as the factors that determine such infections in 99 male sexual partners of women with CIN. The genitalia of the males were physically examined and subjected to peniscopy for the collection of scrapings which were subjected to the polymerase chain reaction and restriction fragment length polymorphism to detect HPV. Of the 99 males sampled, 54 (54.5%) were positive for HPV DNA, 24% of whom presented normal peniscopy, 28% presented evident clinical lesions and 48% isolated lesions consistent with subclinical infection. In the HPV-negative group, 53% showed normal peniscopy, 4% presented evident clinical lesions and 42% isolated lesions consistent with subclinical infection. The study detected a statistically significant association (P < 0.02, Pearson chi-square test) between HPV infection and both the mean number of sexual partners which a male had during his life and the mean number of sexual partners in the year prior to testing. Viral types 6 and 11 were most frequently encountered. The study shows that infection with HPV was frequent in male sexual partners of women with CIN.