The Cerato-Platanin EPL2 from Trichoderma reesei Is Not Directly Involved in Cellulase Formation but in Cell Wall Remodeling.

Trichoderma reesei is a saprophytic fungus that produces large amounts of cellulases and is widely used for biotechnological applications. Cerato-platanins (CPs) are a family of proteins universally distributed among Dikarya fungi and have been implicated in various functions related to fungal physiology and interaction with the environment. In T. reesei, three CPs are encoded in the genome: Trire2_111449, Trire2_123955, and Trire2_82662. However, their function is not fully elucidated. In this study, we deleted the Trire2_123955 gene (named here as epl2) in the wild-type QM6aΔtmus53Δpyr4 (WT) strain and examined the behavior of the Δepl2 strain compared with WT grown for 72 h in 1% cellulose using RNA sequencing. Of the 9143 genes in the T. reesei genome, 760 were differentially expressed, including 260 only in WT, 214 only in Δepl2, and 286 in both. Genes involved in oxidative stress, oxidoreductase activity, antioxidant activity, and transport were upregulated in the Δepl2 mutant. Genes encoding cell wall synthesis were upregulated in the mutant strain during the late growth stage. The Δepl2 mutant accumulated chitin and glucan at higher levels than the parental strain and was more resistant to cell wall stressors. These results suggest a compensatory effect in cell wall remodeling due to the absence of EPL2 in T. reesei. This study is expected to contribute to a better understanding of the role of the EPL2 protein in T. reesei and improve its application in biotechnological fields.

An Analytics Environment Architecture for Industrial Cyber-Physical Systems Big Data Solutions.

The architecture design of industrial data analytics system addresses industrial process challenges and the design phase of the industrial Big Data management drivers that consider the novel paradigm in integrating Big Data technologies into industrial cyber-physical systems (iCPS). The goal of this paper is to support the design of analytics Big Data solutions for iCPS for the modeling of data elements, predictive analysis, inference of the key performance indicators, and real-time analytics, through the proposal of an architecture that will support the integration from IIoT environment, communications, and the cloud in the iCPS. An attribute driven design (ADD) approach has been adopted for architectural design gathering requirements from smart production planning, manufacturing process monitoring, and active preventive maintenance, repair, and overhaul (MRO) scenarios. Data management drivers presented consider new Big Data modeling analytics techniques that show data is an invaluable asset in iCPS. An architectural design reference for a Big Data analytics architecture is proposed. The before-mentioned architecture supports the Industrial Internet of Things (IIoT) environment, communications, and the cloud in the iCPS context. A fault diagnosis case study illustrates how the reference architecture is applied to meet the functional and quality requirements for Big Data analytics in iCPS.

Child maltreatment reports and Child Protection Service responses during COVID-19: Knowledge exchange among Australia, Brazil, Canada, Colombia, Germany, Israel, and South Africa.

BACKGROUND: COVID-19 has become a worldwide pandemic impacting child protection services (CPSs) in many countries. With quarantine and social distancing restrictions, school closures, and recreational venues suspended or providing reduced access, the social safety net for violence prevention has been disrupted significantly. Impacts include the concerns of underreporting and increased risk of child abuse and neglect, as well as challenges in operating CPSs and keeping their workforce safe. OBJECTIVE: The current discussion paper explored the impact of COVID-19 on child maltreatment reports and CPS responses by comparing countries using available population data. METHOD: Information was gathered from researchers in eight countries, including contextual information about the country's demographics and economic situation, key elements of the CPS, and the CPS response to COVID-19. Where available, information about other factors affecting children was also collected. These data informed a discussion about between-country similarities and differences. RESULTS: COVID-19 had significant impact on the operation of every CPS, whether in high- income or low-income countries. Most systems encountered some degree of service disruption or change. Risk factors for children appeared to increase while there were often substantial deficits in CPS responses, and in most countries there was at a temporary decrease in CM reports despite the increased risks to children. CONCLUSIONS: The initial data presented and discussed among the international teams pointed to the way COVID-19 has hampered CPS responses and the protection of children more generally in most jurisdictions, highlighting that children appear to have been at greater risk for maltreatment during COVID-19.

Comparison of the performance of four staging systems in determining the prognosis of breast cancer among women undergoing neoadjuvant chemotherapy.

PURPOSE: Different tumor-related factors have been proposed to assess the risk of disease progression and death in women undergoing neoadjuvant breast cancer chemotherapy. Recently, besides the classical pre-treatment clinical stage (CS) and post-treatment pathologic stage (PS), estrogen receptor status and histologic grade (CPS + EG score) and HER2 results (Neo-Bioscore) have also been added to this suite of staging systems, generating new scores. The present study aims to compare the performance of these four staging systems, namely CS, PS, CPS + EG and Neo-Bioscore, in the prognosis of breast cancer in women undergoing neoadjuvant chemotherapy. METHODS: This study comprises a retrospective cohort study of female breast cancer patients diagnosed at the Brazilian National Cancer Institute, Brazil from January 2013 to December 2015. A descriptive analysis of patient characteristics was conducted, and Kaplan-Meier curves, a Cox proportional hazard analysis and Receiver Operating Characteristic (ROC) curves were developed according to the assessed staging system scores. RESULTS: A total of 803 patients were eligible for this study. Most were under 65 years old (88.0%), presented advanced tumors (clinical stage ≥ IIB 77.1%), with positive estrogen receptor (71.2%) and negative HER2 (75.7%) results. During the follow-up, 172 patients (21.4%) evolved to death. A statistical difference (p < 0.001) was observed between 5 year disease-free survival and 5 year overall survival rates according to the PS, CPS + EG and Neo-Bioscore staging systems. CONCLUSION: The PS, CPS + EG and Neo-Bioscore staging systems were proven to be equivalent to predict the prognosis of patients undergoing neoadjuvant chemotherapy.

Leader-Following Consensus and Formation Control of VTOL-UAVs with Event-Triggered Communications †.

This article presents the design and implementation of an event-triggered control approach, applied to the leader-following consensus and formation of a group of autonomous micro-aircraft with capabilities of vertical take-off and landing (VTOL-UAVs). The control strategy is based on an inner-outer loop control approach. The inner control law stabilizes the attitude and position of one agent, whereas the outer control follows a virtual leader to achieve position consensus cooperatively through an event-triggered policy. The communication topology uses undirected and connected graphs. With such an event-triggered control, the closed-loop trajectories converge to a compact sphere, centered in the origin of the error space. Furthermore, the minimal inter-sampling time is proven to be below bounded avoiding the Zeno behavior. The formation problem addresses the group of agents to fly in a given shape configuration. The simulation and experimental results highlight the performance of the proposed control strategy.

Dataset of cathepsin L-like CP inhibition of Naegleria fowleri and Acanthamoeba castellanii by ppTvCP4r from Trichomonas vaginalis.

The recombinant TvCP4 prepro region (ppTvCP4r) acts as an exogenous inhibitor of cathepsin L-like CPs from Trichomonas vaginalis (Cárdenas-Guerra et al., 2015 [1]). Here, we present the dataset of the trichomonad ppTvCP4r inhibitory effect against the CP proteolytic activities from other microorganisms, such as Naegleria fowleri and Acanthamoeba castellanii free-living amoeba. The proteolytic activity inhibition of total crude extracts (TCEs) of N. fowleri and A. castellanii was determined and recorded using a fluorogenic substrate specific for cathepsin L CPs without or with a ppTvCP4r treatment at different concentrations and pH.

Characterization of Actinobacillus pleuropneumoniae field strains antigenically related to the 3-6-8-15 group from diseased pigs in Japan and Argentina / Caracterización de cepas de Actinobacillus pleuropneumoniae antigénicamente relacionados al grupo 3-6-8-15 obtenidos de cerdos infectados naturalmente en Japón y Argentina

The objectives of this study were to determine the serovar of a collection of Actinobacillus pleuropneumoniae strains within the 3-6-8-15 cross-reacting group and to analyze their phenotypic and genetic properties. Based on the serological tests, forty-seven field strains of Actinobacillus pleuropneumoniae isolated from lungs with pleuropneumonia lesions in Japan and Argentina were found to be serovars belonging to the 3-6-8-15 cross-reacting group. By using a capsule loci-based PCR, twenty-nine (96.7%) and one (3.3%) from Japan were identified as serovars 15 and 8, respectively, whereas seventeen (100%) from Argentina were identified as serovar 8. The findings suggested that serovars 8 and 15 were prevalent within the 3-6-8-15 cross-reacting group, in Argentina and Japan, respectively. Phenotypic analyses revealed that the protein patterns observed on SDS-PAGE and the lipopolysaccharide antigen detected by immunoblotting of the reference and field strains of serovars 8 and 15 were similar to each other. Genetic (16S rDNA, apxIIA, apxIIIA, cps, cpx genes, apx and omlA patterns) analyses revealed that the apxIIA and apxIIIA genes of the field strains of serovars 8 and 15 were similar to those of the reference strains of serovars 3, 4, 6, 8 and 15. The results obtained in the present study may be useful for the development of more effective vaccines against disease caused by A. pleuropneumoniae by including the homologous antigens to the most prevalent serovars in specific geographical areas.

Los objetivos del presente estudio fueron determinar el serovar de una colección de cepas de Actinobacillus pleuropneumoniae pertenecientes al grupo 3, 6, 8, 15 de reacciones cruzadas y analizar sus propiedades fenotípicas y genéticas. En base a técnicas serológicas se determinó que cuarenta y siete cepas de A. pleuropneumoniae aisladas a partir de pulmones con lesiones de pleuroneumonía en Japón y Argentina pertenecen al grupo 3, 6, 8, 15. Mediante el uso de PCR basado en locus capsulares, veintinueve (96.7%) y una (3.3%) de los aislados japoneses fueron identificados como serovar 15 y 8 respectivamente, mientras que diecisiete (100%) de los aislados argentinos resultaron pertenecer al serotipo 8. Este hallazgo sugirió que los serovares 8 y 15 fueron los prevalentes dentro del grupo 3, 6, 8, 15 en Japón y Argentina, respectivamente. El análisis fenotípico reveló que los perfiles proteicos determinados por SDS-PAGE, y de antígenos lipopolisacáridos estudiados por inmunoblot, de las cepas de referencia y de campo de los serovares 8 y 15 fueron similares entre sí. El análisis genético (Í6S rDNA, apxIIA, apxIIA, cps, genes cpx, apx y los perfiles omlA) reveló que los genes apxIIA y apxIIIA de las cepas de campo de los serovares 8 y 15 fueron similares a sus homólogos de las cepas de referencia de los serovares 3, 4, 6, 8 y 15. Los resultados obtenidos en el presente estudio pueden ser útiles para el desarrollo de vacunas más efectivas contra la enfermedad causada por A. pleuropneumoniae, al posibilitar incluir antígenos homólogos a los serovares prevalentes en las áreas geográficas de interés.

Characterization of Actinobacillus pleuropneumoniae field strains antigenically related to the 3-6-8-15 group from diseased pigs in Japan and Argentina.

The objectives of this study were to determine the serovar of a collection of Actinobacillus pleuropneumoniae strains within the 3-6-8-15 cross-reacting group and to analyze their phenotypic and genetic properties. Based on the serological tests, forty-seven field strains of Actinobacillus pleuropneumoniae isolated from lungs with pleuropneumonia lesions in Japan and Argentina were found to be serovars belonging to the 3-6-8-15 cross-reacting group. By using a capsule loci-based PCR, twenty-nine (96.7%) and one (3.3%) from Japan were identified as serovars 15 and 8, respectively, whereas seventeen (100%) from Argentina were identified as serovar 8. The findings suggested that serovars 8 and 15 were prevalent within the 3-6-8-15 cross-reacting group, in Argentina and Japan, respectively. Phenotypic analyses revealed that the protein patterns observed on SDS-PAGE and the lipopolysaccharide antigen detected by immunoblotting of the reference and field strains of serovars 8 and 15 were similar to each other. Genetic (16S rDNA, apxIIA, apxIIIA, cps, cpx genes, apx and omlA patterns) analyses revealed that the apxIIA and apxIIIA genes of the field strains of serovars 8 and 15 were similar to those of the reference strains of serovars 3, 4, 6, 8 and 15. The results obtained in the present study may be useful for the development of more effective vaccines against disease caused by A. pleuropneumoniae by including the homologous antigens to the most prevalent serovars in specific geographical areas.

Symptoms of Daytime Sleepiness and Sleep Apnea in Liver Cirrhosis Patients.

Background/propose. Sleep disturbance and excessive daytime sleepiness (EDS) have been reported in patients with hepatic cirrhosis with no hepatic encephalopathy (HE). The objective of this study was to evaluate daytime sleepiness and risk of obstructive sleep apnea (OSA) among liver cirrhosis patients. MATERIAL AND METHODS: A cross-sectional study was conducted at King Abdulaziz Medical City (KAMC)-Riyadh over a period of six months, using a structured questionnaire that investigated: 1) Sleep patterns and daytime sleepiness using the Epworth Sleeping Scale (ESS), and 2) The risk for sleep apnea using the Berlin Questionnaire (BQ). We enrolled patients with a confirmed diagnosis of liver cirrhosis who were being followed at the hepatology and pre-liver transplant clinics. RESULTS: We enrolled 200 patients with liver cirrhosis, 57.5% of whom were male. The mean age was 60 (± SD 12.2). The reported prevalence of EDS, OSA, and both EDS and OSA were 29.5%, 42.9%, and 13.6%, respectively. The prevalence of EDS was higher in patients with Hepatitis-C and patients with DM, who experienced short sleep duration. We did not find any association between the severity of liver disease and EDS or OSA as measured by Child-Pugh scores (CPS). CONCLUSIONS: The risk of OSA and EDS is high among liver cirrhosis patients. Those patients with cirrhosis secondary to Hepatitis C are at higher risk of EDS and OSA. Both EDS and OSA affect patients designated as CPS Class A more frequently than patients designated as CPS Class B.

Impact of CPS1 Gene rs7422339 Polymorphism in Argentine Patients With Hyperhomocysteinemia

Abstract Carbamoyl phosphate synthetase 1 (CPS1) is a key gene in the first step of urea cycle and has been correlated with nitric oxide level and vascular smooth muscle activity. A functional single-nucleotide polymorphism C/A at position 4217 in CPS1 (National Center for Biotechnology Information SNP database no. rs7422339, T1405N) was reported to be associated with high homocysteine (Hcy) plasma values. Although genetic variants of methylenetetrahydrofolate reductase (MTHFR) gene are known to influence Hcy concentration, other genetic determinants of Hcy remain largely unknown. The association between the CPS1 rs7422339 and the risk of hyperhomocysteinemia in Latin American populations is unknown. Here, we study this association in 100 patients having hyperhomocysteinemia without MTHFR c.677C>T polymorphism and 100 controls. CPS1 rs7422339 was studied using polymerase chain reaction and enzymatic restriction. Comparisons of the CPS1 rs7422339 genotype distributions revealed a significant difference between groups (P = 2.3 × 10−3). Patients carrying polymorphic allele showed almost 3 times higher risk (odds ratio [OR] = 2.47) of hyperhomocysteinemia than wild-type allele, suggesting that rs7422339 SNP is associated with high Hcy levels in the Argentine population.

Trichocystatin-2 (TC-2): an endogenous inhibitor of cysteine proteinases in Trichomonas vaginalis is associated with TvCP39.

The causal agent of trichomoniasis is a parasitic protist, Trichomonas vaginalis, which is rich in proteolytic activity, primarily carried out by cysteine proteases (CPs). Some CPs are known virulence factors. T. vaginalis also possesses three genes encoding endogenous cystatin-like CP inhibitors. The aim of this study was to identify and characterize one of these CP inhibitors. Using two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS), a cystatin-like peptidase inhibitor dubbed Trichocystatin-2 (TC-2) was identified in the T. vaginalis active degradome in association with TvCP39, a 39kDa CP involved in cytotoxicity. To characterize the TC-2 inhibitor, we cloned and expressed the tvicp-2 gene, purified the recombinant protein (TC-2r), and produced a specific polyclonal antibody (α-TC-2r). This antibody recognized a 10kDa protein band by western blotting. An indirect immunofluorescence assay (IFA) and cell fractionation assays using the α-TC-2r antibody showed that TC-2 was localized in the cytoplasm and lysosomes and that it colocalized with TvCP39. TC-2r showed inhibitory activity against papain, cathepsin-L, and TvCP39 in trichomonad extracts and live parasites but not legumain-like CPs. Live trichomonads treated with TC-2r showed reduced trichomonal cytotoxicity to HeLa cell monolayers in a TC-2r-concentration-dependent manner. In this study, we identified and characterized an endogenous cystatin-like inhibitor in T. vaginalis, TC-2, which is associated with TvCP39 and appears to regulate the cellular damage caused by T. vaginalis.

Calcium carbonate precipitation by heterotrophic bacteria isolated from biofilms formed on deteriorated ignimbrite stones: influence of calcium on EPS production and biofilm formation by these isolates.

Heterotrophic CaCO3-precipitating bacteria were isolated from biofilms on deteriorated ignimbrites, siliceous acidic rocks, from Morelia Cathedral (Mexico) and identified as Enterobacter cancerogenus (22e), Bacillus sp. (32a) and Bacillus subtilis (52g). In solid medium, 22e and 32a precipitated calcite and vaterite while 52g produced calcite. Urease activity was detected in these isolates and CaCO3 precipitation increased in the presence of urea in the liquid medium. In the presence of calcium, EPS production decreased in 22e and 32a and increased in 52g. Under laboratory conditions, ignimbrite colonization by these isolates only occurred in the presence of calcium and no CaCO3 was precipitated. Calcium may therefore be important for biofilm formation on stones. The importance of the type of stone, here a siliceous stone, on biological colonization is emphasized. This calcium effect has not been reported on calcareous materials. The importance of the effect of calcium on EPS production and biofilm formation is discussed in relation to other applications of CaCO3 precipitation by bacteria.

A prospective study of sudden unexpected infant death after reported maltreatment.

OBJECTIVE: To examine whether infants reported for maltreatment face a heightened risk of sudden infant death syndrome (SIDS) and other leading causes of sudden unexpected infant death (SUID). STUDY DESIGN: Linked birth and infant death records for all children born in California between 1999 and 2006 were matched to administrative child protection data. Infants were prospectively followed from birth through death or 1 year of age. A report of maltreatment was modeled as a time-varying covariate; risk factors at birth were included as baseline covariates. Multivariable competing risk survival models were used to estimate the adjusted relative hazard of postneonatal SIDS and other SUID. RESULTS: A previous maltreatment report emerged as a significant predictor of SIDS and other SUID. After adjusting for baseline risk factors, the rate of SIDS was more than 3 times as great among infants reported for possible maltreatment (hazard ratio: 3.22; 95% CI: 2.66, 3.89). CONCLUSION: Infants reported to child protective services have a heightened risk of SIDS and other SUID. Targeted services and improved communication between child protective services and the pediatric health care community may enhance infant well-being and reduce risk of death.

Predictors of screening and injury in contacts of physically abused children.

OBJECTIVE: To determine rates of screening in contacts of children evaluated for physical abuse, and the relationship of clinical characteristics to screening recommendation and completion and injury identification. STUDY DESIGN: This is a planned secondary analysis of a prospective study of 1918 contacts of 1196 children referred for subspecialty abuse consultation in 20 US centers. We used multivariable logistic models to determine the relationship of index child characteristics, contact child characteristics, and shared characteristics to screening and injury identification. RESULTS: We identified injuries or disclosures of abuse in 180 (9.4%) contacts. Recommended screening was omitted in >20% of subjects for each screening modality. At least 1 screening test was more likely to be completed in contacts of index children of non-White race or Hispanic ethnicity (OR 1.45, 95% CI 1.13-1.87), with abuse-specific injuries (OR 2.15, 95% CI 1.63-2.83), with a confession (OR 2.18, 95% CI 1.17-4.07), when the history changed (OR 1.65, 95% CI 1.05-2.61), when an occult injury was found by imaging in the index child (OR 1.84, 95% CI 1.39-2.43), and when families lacked private insurance (OR 1.63, 95% CI 1.15-2.31). CONCLUSION: Completion of screening recommended for contacts of potentially abused children is relatively poor, despite high risk of injury. Several clinical and demographic factors were associated with increased contact screening.

Microbiological quality of finger food and snacks / Qualidade microbiológica de lanches e salgados

Although finger food is convenient, it may be easily contaminated from the stage of preparation to the moment of consumption. This study aimed at evaluating the microbiological quality of finger food and sandwiches sold in Botucatu SP, Brazil, by following the standards established by the Brazilian Health Surveillance Agency, ANVISA. The analysis was conducted according to APHA. A hundred and twenty-two samples of meat, chicken, shrimp, cheese, and vegetable finger food and sandwiches were tested from August 2008 to March 2009. Seventeen (13.9 %) samples of meat, cheese, vegetables and chicken were in disagreement with the ANVISA standards some of them in more than one parameter. High counting of thermotolerant coliforms and coagulase-positive staphylococci were found in ten (8.2 %) and eight (6.5 %) samples, respectively. Salmonella spp. was detected in two samples (1.6 %). No Bacillus cereus and sulfite-reducing clostridia were isolated. Although only 10 samples (8.2 %) showed pathogenic bacteria contamination, these results are relevant, since they indicate that the population is generally exposed to risks of acquiring foodborne diseases. Thus, the sanitary authorities might implement actions for supervising the quality of the food sold in Botucatu, and to strengthen the food sellers to improve the hygienic conditions and be aware of the risks of food contamination.

Neste trabalho foi avaliada a qualidade microbiológica de salgados e sanduíches comercializados em Botucatu-SP, seguindo-se a legislação em vigor. As análises foram realizadas de acordo com APHA. Foram analisadas 122 amostras de salgados de carne, frango, camarão, queijo e vegetais, e sanduíches no período de agosto/2008 a março/2009. Dezessete (13,9 %) amostras de carne, queijo, vegetais e frango, e em algumas em mais de um parâmetro, estavam em desacordo com a legislação em vigor. Foram detectadas elevadas contagens de coliformes termotolerantes e estafilococos coagulase-positiva, respectivamente, em dez (8,2 %) e oito (6,5 %) amostras. Salmonella spp. foi isolada em duas amostras (1,6 %). As contagens de Bacillus cereus e de Clostrídio Sulfito Redutor não ultrapassaram os padrões da legislação. A maioria dos salgados mostrou resultados dentro dos padrões estabelecidos pela legislação. A presença de bactérias patogênicas como S. aureus e Salmonella spp. foi demonstrada em 10 amostras (8,2 %); e este resultado é relevante, pois indica que a população está exposta a riscos de doenças veiculadas por alimentos. Torna-se necessário colocar em prática a vigilância dos alimentos comercializados em Botucatu, incentivar a melhoria de condições de higiene pelos comerciantes, e ter ciência dos riscos e das implicações da contaminação microbiológica dos alimentos.

Qualidade microbiológica de lanches e salgados / Microbiological quality of finger food and snacks

Although finger food is convenient, it may be easily contaminated from the stage of preparation to themoment of consumption. This study aimed at evaluating the microbiological quality of finger food and sandwiches sold in Botucatu – SP, Brazil, by following the standards established by the Brazilian Health Surveillance Agency, ANVISA. The analysis was conducted according to APHA. A hundred and twenty two samples of meat, chicken, shrimp, cheese, and vegetable finger food and sandwiches were tested fromAugust 2008 to March 2009. Seventeen (13.9 %) samples of meat, cheese, vegetables and chicken were indisagreement with the ANVISA standards — some of them in more than one parameter. High counting of thermotolerant coliforms and coagulase-positive staphylococci were found in ten (8.2 %) and eight(6.5 %) samples, respectively. Salmonella spp. was detected in two samples (1.6 %). No Bacillus cereus andsulfite-reducing clostridia were isolated. Although only 10 samples (8.2 %) showed pathogenic bacteriacontamination, these results are relevant, since they indicate that the population is generally exposedto risks of acquiring foodborne diseases. Thus, the sanitary authorities might implement actions forsupervising the quality of the food sold in Botucatu, and to strengthen the food sellers to improve thehygienic conditions and be aware of the risks of food contamination.

Neste trabalho foi avaliada a qualidade microbiológica de salgados e sanduíches comercializados emBotucatu-SP, seguindo-se a legislação em vigor. As análises foram realizadas de acordo com APHA. Foramanalisadas 122 amostras de salgados de carne, frango, camarão, queijo e vegetais, e sanduíches no períodode agosto/2008 a março/2009. Dezessete (13,9 %) amostras de carne, queijo, vegetais e frango, e em algumasem mais de um parâmetro, estavam em desacordo com a legislação em vigor. Foram detectadas elevadascontagens de coliformes termotolerantes e estafilococos coagulase-positiva, respectivamente, em dez (8,2 %)e oito (6,5 %) amostras. Salmonella spp. foi isolada em duas amostras (1,6 %). As contagens de Bacillus cereuse de Clostrídio Sulfito Redutor não ultrapassaram os padrões da legislação. A maioria dos salgados mostrouresultados dentro dos padrões estabelecidos pela legislação. A presença de bactérias patogênicas comoS. aureus e Salmonella spp. foi demonstrada em 10 amostras (8,2 %); e este resultado é relevante, pois indicaque a população está exposta a riscos de doenças veiculadas por alimentos. Torna-se necessário colocarem prática a vigilância dos alimentos comercializados em Botucatu, incentivar a melhoria de condiçõesde higiene pelos comerciantes, e ter ciência dos riscos e das implicações da contaminação microbiológicados alimentos.

Uso de PCR e sequenciamento do rDNA 16S para identificação de bactérias do gênero Staphylococcus isoladas de mastite bovina / Identification of Staphylococcus strains isolated from bovine mastitis by PCR and 16S rDNA sequencing

O objetivo deste trabalho foi identificar espécies de Staphylococcus (n=100) isoladas de mastite em rebanhos bovinos do Estado de Minas Gerais. Para esta finalidade foram utilizadas reações de PCR empregando oligonucleotídeos iniciadores descritos anteriormente para amplificar genes específicos de S. aureus (femA), S. intermedius (rDNA 16S) e S. hyicus (rDNA 16S-23S) e o sequenciamento do rDNA 16S. De acordo com as reações de PCR, 83 isolados foram identificados como S. aureus, 13 isolados como S. intermedius, dois como S. hyicus e dois isolados não foram identificados. Foram submetidos ao sequenciamento do rDNA 16S seis isolados identificados como S. aureus e os 17 restantes. Os seis isolados identificados como S. aureus confirmaram essa identificação. Dos outros 17 isolados, 13 foram identificados como S. chromogenes e quatro como S. hyicus, com similaridade igual ou superior a 99%. Baseando-se nos resultados da reação de PCR do gene femA e do sequenciamento do rDNA 16S, foram identificados 83 S. aureus, 13 S. chromogenes e quatro S. hyicus. Neste estudo os oligonucleotídeos iniciadores empregados na reação de PCR para S. intermedius não foram específicos, pois amplificaram também S. chromogenes; e os empregados na reação de PCR para S. hyicus não foram sensíveis, pois falharam na identificação de dois isolados de S. hyicus. A identificação definitiva das duas últimas espécies somente foi possível pelo sequenciamento do rDNA 16S.(AU)

The objective of this study was to identify the species of 100 isolates of Staphylococcus from mastitis in dairy cows from herds located in the state of Minas Gerais, Brazil. PCR reactions were carried out using specific primers described previously for S. aureus (femA gene), S. intermedius (16S rDNA) and S. hyicus (16S-23S rDNA spacer region). In addition, products of amplification of variable regions of the 16S rDNA gene of the strains were sequenced. According to the results of the PCR, 83 strains were identified as S. aureus, 13 as S. intermedius, two as S. hyicus and two isolates were not identified. The sequencing of 16S rDNA was applied to 23 strains identified by PCR amplifications: six S. aureus and the strains identified as S. intermedius (n=13), S. hyicus (n=2) or not identified (n=2). The sequencing of 16S rDNA confirmed the six strains as S. aureus. The others 17 strains were identified as S. chromogenes (13 isolates) and S. hyicus (four isolates). Each sample was related to a specie according to the smallest E-value and highest similarity (≥ 99%). The identification of S. hyicus and S. chromogenes was accomplished only by 16S rDNA sequencing.(AU)

Uso de PCR e sequenciamento do rDNA 16S para identificação de bactérias do gênero staphylococcus isoladas de mastite bovina / Identification of staphylococcus strains isolated from bovine mastitis by PCR and 16S rDNA sequencing

O objetivo deste trabalho foi identificar espécies de Staphylococcus (n=100) isoladas de mastite em rebanhos bovinos do Estado de Minas Gerais. Para esta finalidade foram utilizadas reações de PCR empregando oligonucleotídeos iniciadores descritos anteriormente para amplificar genes específicos de S. aureus (femA), S. intermedius (rDNA 16S) e S. hyicus (rDNA 16S-23S) e o sequenciamento do rDNA 16S. De acordo com as reações de PCR, 83 isolados foram identificados como S. aureus, 13 isolados como S. intermedius, dois como S. hyicus e dois isolados não foram identificados. Foram submetidos ao sequenciamento do rDNA 16S seis isolados identificados como S. aureus e os 17 restantes. Os seis isolados identificados como S. aureus confirmaram essa identificação. Dos outros 17 isolados, 13 foram identificados como S. chromogenes e quatro como S. hyicus, com similaridade igual ou superior a 99%. Baseando-se nos resultados da reação de PCR do gene femA e do sequenciamento do rDNA 16S, foram identificados 83 S. aureus, 13 S. chromogenes e quatro S. hyicus. Neste estudo os oligonucleotídeos iniciadores empregados na reação de PCR para S. intermedius não foram específicos, pois amplificaram também S. chromogenes; e os empregados na reação de PCR para S. hyicus não foram sensíveis, pois falharam na identificação de dois isolados de S. hyicus. A identificação definitiva das duas últimas espécies somente foi possível pelo sequenciamento do rDNA 16S.

The objective of this study was to identify the species of 100 isolates of Staphylococcus from mastitis in dairy cows from herds located in the state of Minas Gerais, Brazil. PCR reactions were carried out using specific primers described previously for S. aureus (femA gene), S. intermedius (16S rDNA) and S. hyicus (16S-23S rDNA spacer region). In addition, products of amplification of variable regions of the 16S rDNA gene of the strains were sequenced. According to the results of the PCR, 83 strains were identified as S. aureus, 13 as S. intermedius, two as S. hyicus and two isolates were not identified. The sequencing of 16S rDNA was applied to 23 strains identified by PCR amplifications: six S. aureus and the strains identified as S. intermedius (n=13), S. hyicus (n=2) or not identified (n=2). The sequencing of 16S rDNA confirmed the six strains as S. aureus. The others 17 strains were identified as S. chromogenes (13 isolates) and S. hyicus (four isolates). Each sample was related to a specie according to the smallest E-value and highest similarity (≥ 99%). The identification of S. hyicus and S. chromogenes was accomplished only by 16S rDNA sequencing.

Uso masivo de la coproparasitoscopía con FAF / Masive use of coproparasitoscopy with FAF

El uso de solución preservadora y fijadora (FAF) de formas parásitas en la coproparasitoscopía de rutina para el diagnóstico de parasitosis intestinales, es considerado en este trabajo como la parte escencial para la realización de encuestas y trabajos de campo, cuando se desea tener una mayor confiabilidad, así como cuando se tienen que transportar lals muestras a un laboratorio de concentración. Su uso continuo ahorra material y equipo, y es adecuado para el procesamiento de los exámenes que más comunmente solicita el cliente