Environmental assessment and durability performance of cement mortar incorporating sugarcane vinasse in replacement of water.

Sugarcane vinasse wastewater (SVW) is one of the most voluminous waste generated in the ethanol industry and usually applied in fertigation. It is characterized by presenting high COD and BOD; thus, continued disposal of vinasse results in negative environmental impacts. In this paper, we investigated the potential of SVW in replacement of water in mortar, rethinking about reuse of effluent, reduction of pollutants in the environment, and water consumption in civil construction. Mortar composites with 0, 20, 40, 60, 80, and 100% of water replaced by SVW were studied in order to determine the optimum content. Mortars with 60 to 100% of SVW result in improved workability and reduction in water demand. The mortars with 20, 40, and 60% SVW resulted in satisfactory mechanical properties, i.e., similar to the control mortar. However, XRD analysis of cement pastes showed that the SVW causes a delay in CH formation, reaching mechanical strength after 28 days. Durability tests results showed that SVW contributes to the mortar becoming more impermeable; therefore, less susceptible to weathering. This study provides an important evaluation of the potential of SVW for application in civil construction, indicating relevant results for replacement of water by liquid wastes in cementitious composites and reduction the use of natural resources.

Escherichia coli Strains Responsible for Cystitis in Female Pediatric Patients with Normal and Abnormal Urinary Tracts Have Different Virulence Profiles.

The role of uropathogenic Escherichia coli (UPEC) in colonization and infection of female patients with anatomical and functional abnormalities of the urinary system is elusive. In this study, the phenotype, genotype and the phylogeny of UPEC strains isolated from the urine of pediatric female patients with cystitis of normal and abnormal urinary tract were determined. Multiplex PCR results demonstrated that 86% of the strains isolated from female patients with normal urinary tract (NUT), belonged to the phylo-groups B2 and D. Their prevalence decreased to 23% in strains isolated from patients with abnormal urinary tract (AUT). More of the isolates from AUT patients produced a biofilm on polystyrene and polyvinyl chloride (PVC), adhered to epithelial cells, and encoded pap and sfa genes than strains isolated from female patients with NUT. In contrast, a higher number of hemolysin-producing strains with serogroups associated with UPEC were isolated from patients with NUT. In summary, the results suggest that cystitis in female patients with NUT is associated with ExPEC, whereas cystitis in female patients with AUT is associated with pathogenic intestinal E. coli strains that have acquired the ability to colonize the bladder.

Efficacy of hydroxychloroquine in the prevention of thromboembolic events: A systematic review and meta-analysis.

OBJECTIVE: To estimate the risk ratio (RR) of thromboembolic events in chloroquine and hydroxychloroquine users compared to non-users. METHODS: We systematically reviewed randomized controlled trials (RCTs), using MEDLINE and EMBASE databases from inception to the present, reporting thromboembolic events in chloroquine and hydroxychloroquine users compared to non-users. Four authors independently screened all the records obtained through our search strategy and later revised the selected full-text articles for eligibility, according to our inclusion criteria. The same four authors independently extracted relevant data through a customized data collection form while two other authors assessed the quality of the included RCTs using the Cochrane risk-of-bias tool (Version 2.0). All the disagreements were resolved through discussions among the authors. We calculated the risk ratio (RR) and its respective standard error of developing thromboembolic events in hydroxychloroquine users and non-users for each individual study and pooled the results using a random effects model meta-analysis. We assessed Heterogeneity using the Tau2 and I2, and publication bias using funnel plotting and Egger's regression. The protocol for this systematic review is registered at the PROSPERO database (CRD42021247902). RESULTS: Thirteen RCTs met our eligibility criteria and were included in our analysis (2663 patients). We found that hydroxychloroquine-no study on chloroquine was found-reduced the risk of thromboembolic events by 49% (RR 0.51[IC 95% 0.31-0.84]) with a medium heterogeneity (I2 = 67% and T2 = 0.4948). We did find some asymmetry in the inspection of the funnel plot, which was ruled out through an Egger's regression (p-value = 0.1025). CONCLUSION: Our data reinforce the idea that hydroxychloroquine reduces the risk of thromboembolic events.

Effects of orchiectomy and testosterone replacement therapy on redox balance and salivary gland function in Wistar rats.

The objective of this study was to investigate the effects of orchiectomy (ORX) and testosterone replacement therapy (TRT) on redox balance and function of salivary glands. Forty-five young adult male Wistar rats (3 months old) were either castrated bilaterally or underwent fictitious surgery (SHAM) and were subsequently distributed into 3 groups: SHAM, ORX, and TRT (castrated rats that received an intramuscular injection of testosterone cypionate 10 mg/kg/weekly). All treatments started 4 weeks after castration (4 months old) and lasted 4 weeks (5 months old). At the end of treatment, pilocarpine-induced salivary secretion was collected to analyze salivary flow rate and biochemistry composition, and the parotid (PG) and submandibular (SMG) glands were sampled for redox balance markers and histomorphometric analyses. ORX increased salivary flow rate, calcium, phosphate, and chloride, and decreased total protein and amylase, while not changing the salivary buffer capacity, pH, sodium, and potassium compared to SHAM. TRT restored all salivary parameters to SHAM values. ORX increased oxidative lipid and protein damage, total antioxidant capacity, and uric acid in both salivary glands compared to SHAM. Superoxide dismutase, catalase, and glutathione peroxidase activities were greater only in the SMG of the ORX group in relation to SHAM. ORX decreased duct and acini area, while increasing connective tissue in the PG. On the other hand, ORX reduced duct area and increased acini area in the SMG compared to SHAM. TRT restored the redox balance and histomorphometric parameters to close to SHAM values in both salivary glands. Orchiectomy-induced salivary gland dysfunction was characterized by an increase in the salivary flow rate and changes in the secretion of total protein, amylase, and electrolytes, which are key factors, considered important for maintaining oral health status. To sum up, orchiectomy impaired the redox balance of the salivary glands. Our results also showed that TRT reversed the oxidative damage, morphological alterations, and salivary gland dysfunction induced by orchiectomy. Therefore, these results suggest an important action of testosterone on the redox balance and secretory ability of salivary glands.

EHEC O111:H8 strain and norovirus GII.4 Sydney [P16] causing an outbreak in a daycare center, Brazil, 2019.

BACKGROUND: This study describes the investigation of an outbreak of diarrhea, hemorrhagic colitis (HC), and hemolytic uremic syndrome (HUS) at a daycare center in southeastern Brazil, involving fourteen children, six staff members, six family members, and one nurse. All bacterial and viral pathogens detected were genetically characterized. RESULTS: Two isolates of a strain of enterohemorrhagic Escherichia coli (EHEC) serotype O111:H8 were recovered, one implicated in a case of HUS and the other in a case of uncomplicated diarrhea. These isolates had a clonal relationship of 94% and carried the stx2a and eae virulence genes and the OI-122 pathogenicity island. The EHEC strain was determined to be a single-locus variant of sequence type (ST) 327. EHEC isolates were resistant to ofloxacin, doxycycline, tetracycline, ampicillin, and trimethoprim-sulfamethoxazole and intermediately resistant to levofloxacin and ciprofloxacin. Rotavirus was not detected in any samples, and norovirus was detected in 46.7% (14/30) of the stool samples, three of which were from asymptomatic staff members. The noroviruses were classified as the recombinant GII.4 Sydney [P16] by gene sequencing. CONCLUSION: In this outbreak, it was possible to identify an uncommon stx2a + EHEC O111:H8 strain, and the most recent pandemic norovirus strain GII.4 Sydney [P16]. Our findings reinforce the need for surveillance and diagnosis of multiple enteric pathogens by public health authorities, especially during outbreaks.

Genotypic and phenotypic characterization of enteroaggregative Escherichia coli (EAEC) isolates from diarrheic children: An unresolved diagnostic paradigm exists.

OBJECTIVES: The enteroaggregative Escherichia coli (EAEC) has been one of the most intriguing emerging bacterial pathogens in children that occur both in developing countries and the industrial world. Although various phenotypic and genotypic based protocols have been suggested for diagnosis of EAEC, they are not conclusive or practical to be used in most clinical laboratories. MATERIALS AND METHODS: In this study, we analyzed and compared 36 typical EAEC strains (aggR-positive) by various genotypic and phenotypic methods. RESULTS: Briefly, pCVD432 was detected in all of isolates along with aggR, then it was followed by other virulence genes including app, astA, aggA, and pet genes in 32 (88.8%), 21 (58.3%), 9 (25%), and 2 (5.5%) isolates, respectively. Biofilm was formed by 34 (94.4%) isolates, while only 26 (72.2%) isolates showed an aggregative adherence pattern to HEp-2 cells. CONCLUSION: The genetic and phenotypic features of EAEC were highly inconsistent, which may have considerable diagnostic implications. The variations in the virulence genes, phenotypic characteristics, and genetic profiles among the EAEC isolates again emphasized the genetic heterogeneity of this emerging pathotype. Biofilm formation may be an important phenotypic virulence property of this pathotype, especially in strains with the aggR-pCVD432-aap-astA profile.

Development and Validation of Shiga Toxin-Producing Escherichia coli Immunodiagnostic Assay.

Shiga toxin (Stx)-producing Escherichia coli (STEC) and its subgroup enterohemorrhagic E. coli are important pathogens involved in diarrhea, which may be complicated by hemorrhagic colitis and hemolytic uremic syndrome, the leading cause of acute renal failure in children. Early diagnosis is essential for clinical management, as an antibiotic treatment in STEC infections is not recommended. Previously obtained antibodies against Stx1 and Stx2 toxins were employed to evaluate the sensitivity and specificity of the latex Agglutination test (LAT), lateral flow assay (LFA), and capture ELISA (cEIA) for STEC detection. The LAT (mAb Stx1 plus mAb stx2) showed 99% sensitivity and 97% specificity. Individually, Stx1 antibodies showed 95.5% and 94% sensitivity and a specificity of 97% and 99% in the cEIA and LFA assay, respectively. Stx2 antibodies showed a sensitivity of 92% in both assays and a specificity of 100% and 98% in the cEIA and LFA assay, respectively. These results allow us to conclude that we have robust tools for the diagnosis of STEC infections.

Occurrence and antimicrobial resistance of E. coli non-O157 isolated from beef in Mato Grosso, Brazil.

Shiga toxin-producing Escherichia coli (STEC) is an important public health concern pathogen, as it produces two toxins, Stx1 and Stx2, with cytotoxic capacity. In addition, STEC strains are frequently involved in food outbreaks worldwide, leading to public health challenges and economic losses. In this context, the occurrence and antimicrobial resistance profile of the STEC isolated from fresh beef produced in Mato Grosso, Brazil, were estimated. One hundred seven retail beef under vacuum-packaged produced by 13 different slaughterhouses were submitted to microbiological, molecular, and antimicrobial resistance analyses. STEC occurrence in beef was of 4.67%, and five strains presented the stx2 gene. The O111 serogroup, reported in several outbreak cases worldwide, was detected, and other serotypes (O8:H20, O22:H16, and O141:H49) were also isolated. All isolated strains displayed sensitivity to 12 antibiotics, except for two strains, which where streptomycin-resistant. The presence of STEC in retail beef samples indicates public health risks with significant economic impact throughout the retail beef chain.

Novel Hybrid of Typical Enteropathogenic Escherichia coli and Shiga-Toxin-Producing E. coli (tEPEC/STEC) Emerging From Pet Birds.

Exotic psittacine birds have been implicated as reservoir of diarrheagenic Escherichia coli (E. coli), including enteropathogenic E. coli (EPEC) and Shiga-toxin producing E. coli (STEC). Here, we present a genotypic and phenotypic characterization of typical EPEC/STEC hybrid strains isolated from exotic psittacine birds. The strains were positive for eae, bfpA, and stx2f genes, belong to serotype O137:H6 and ST2678. Two strains were subject to whole genome sequencing, confirming the presence of the virulence factors of both E. coli pathotypes. Phenotypical in vitro tests confirmed their ability to adhere to HeLa cells and cause cytotoxicity to Vero cells. The rabbit ileal loop assays showed the attaching and effacing lesion, in addition to inflammatory process and overproduction of intestinal mucus. This is the first report of hybrid typical EPEC/STEC (O137:H6/ST2678) strains isolated from companion psittacine birds and the results suggest zoonotic risks.

Shigatoxigenic and atypical enteropathogenic Escherichia coli in fish for human consumption

ABSTRACT Shigatoxigenic and enteropathogenic Escherichia coli with virulence and multidrug resistance profile were isolated from Nile tilapia. This study finding is of great importance to public health because they help understand this pathogen epidemiology in fish and demonstrate how these animals can transmit E. coli related diseases to humans.

Shigatoxigenic and atypical enteropathogenic Escherichia coli in fish for human consumption.

Shigatoxigenic and enteropathogenic Escherichia coli with virulence and multidrug resistance profile were isolated from Nile tilapia. This study finding is of great importance to public health because they help understand this pathogen epidemiology in fish and demonstrate how these animals can transmit E. coli related diseases to humans.

Large-scale evaluation of a rapid diagnostic test for diarrhea caused by enterotoxigenic Escherichia coli targeting the heat-labile toxin.

We standardized an immunochromatographic test (IC) for heat-labile toxin I (LT-I) detection using LT-I antibodies and a specific platform containing the apparatus for application, assembly and cutting. IC detected as little as 62.5ng/mL of purified LT-I toxin and presented 91% sensitivity, 99.5% specificity and 96.0% accuracy, thereby proving to be an excellent point-of-care test for the diagnosis of enterotoxigenic E. coli infection in low-income countries.

Assessment of Consumer Exposure to Salmonella spp., Campylobacter spp., and Shiga Toxin-Producing Escherichia coli in Meat Products at Retail in the City of Sao Paulo, Brazil.

Meat products may be vehicles of bacterial pathogens to humans, and Salmonella spp., Campylobacter spp., and Shiga toxin-producing Escherichia coli (STEC) are the most relevant. The aim of this study was to generate data on prevalence of these three pathogens in 552 samples of meat products (hot dogs, pork sausages, raw ground beef, and raw chicken legs) sold at retail in the city of Sao Paulo, Brazil. Salmonella spp. was detected in 5.8% (32/552) of samples, comprising pork sausages 62.5% (20/32) and chicken legs 37.5% (12/32). The counts of Salmonella spp. were low, ranging from < 0.3 to 9.3 × 10 most probable number per gram and the most frequent serovars were Salmonella Typhimurium (28.1%), Salmonella I 4,[5],12:i:- (15.6%), Salmonella Enteritidis (12.5%), Salmonella Derby, and Salmonella Brandenburg (9.4%). Campylobacter spp. was detected in 33 samples (6.0%), comprising chicken legs (82%) and ground beef (18%). All samples were negative for STEC. These results suggest that meat products when subjected to inadequate cooking and/or cross-contamination with other products ready for consumption can lead to occurrence of outbreaks, highlighting the risks associated with them.

Is the microagglutination test (MAT) good for predicting the infecting serogroup for leptospirosis in Brazil?

Leptospirosis is a zoonotic infection caused by pathogenic members of the genus Leptospira spp. Knowledge of the prevalent serovars and their maintenance hosts is essential to understand the disease. The aim of this study was to evaluate the ability of serology by the microscopic agglutination test (MAT) to predict the serogroups compared with results of identification of leptospires in São Paulo, Brazil. MAT correctly assigned the serogroup of the infecting isolate in 49/52 cases (94.23%). The serogroup Icterohaemorrhagiae was the predominant serogroup (88.46%). This study showed the usefulness of the MAT to correctly identify the infecting serogroup with a good overall agreement between the serologically-identified infecting serogroup and by identification of the isolate and can be used in epidemiological surveys in São Paulo. However, it should be complemented by the identification of Leptospira isolates.

Genetic diversity and virulence potential of shiga toxin-producing Escherichia coli O113:H21 strains isolated from clinical, environmental, and food sources.

Shiga toxin-producing Escherichia coli strains of serotype O113:H21 have caused severe human diseases, but they are unusual in that they do not produce adherence factors coded by the locus of enterocyte effacement. Here, a PCR microarray was used to characterize 65 O113:H21 strains isolated from the environment, food, and clinical infections from various countries. In comparison to the pathogenic strains that were implicated in hemolytic-uremic syndrome in Australia, there were no clear differences between the pathogens and the environmental strains with respect to the 41 genetic markers tested. Furthermore, all of the strains carried only Shiga toxin subtypes associated with human infections, suggesting that the environmental strains have the potential to cause disease. Most of the O113:H21 strains were closely related and belonged in the same clonal group (ST-223), but CRISPR analysis showed a great degree of genetic diversity among the O113:H21 strains.

Set of virulence genes and genetic relatedness of O113 : H21 Escherichia coli strains isolated from the animal reservoir and human infections in Brazil.

Escherichia coli strains of serotype O113 : H21 are commonly described as belonging to a Shiga toxin (Stx)-producing E. coli (STEC) pathotype worldwide. Albeit this STEC serotype is frequently identified among cattle and other domestic animals, to the best of our knowledge no human infections associated with STEC O113:H21 have been registered in Brazil to date. Here, we report the virulence profile and genetic relatedness of a collection of O113:H21 E. coli strains mainly isolated from the animal reservoir aimed at determining their potential as human pathogens. The strains from the animal reservoir (n=34) were all classified as STEC, whereas the few isolates recovered so far from human diarrhoea (n=3) lacked stx genes. Among the STEC, the stx2d-activatable gene was identified in 85% of the strains that also carried lpfAO113, iha, saa, ehxA, subAB, astA, cdt-V, espP, espI and epeA; the human strains harboured only lpfAO113, iha and astA. All the strains except one, isolated from cattle, were genetically classified as phylogenetic group B1. High mass plasmids were observed in 25 isolates, but only in the STEC group were these plasmids confirmed as the STEC O113 megaplasmid (pO113). Many closely related subgroups (more than 80% similarity) were identified by PFGE, with human isolates clustering in a subgroup separate from most of the animal isolates. In conclusion, potentially pathogenic O113:H21 STEC isolates carrying virulence markers in common with O113:H21 clones associated with haemolytic uraemic syndrome cases in other regions were demonstrated to occur in the natural reservoir in our settings, and therefore the risk represented by them to public health should be carefully monitored.

Subtilase cytotoxin-encoding subAB operon found exclusively among Shiga toxin-producing Escherichia coli strains.

The presence of subAB was investigated for 3,453 Escherichia coli strains of various pathogenic categories. The occurrence of other virulence genes in subAB-positive strains was investigated. The subAB operon was detected among some Shiga toxin-producing E. coli (STEC) serotypes devoid of eae and carrying ehxA. Most subAB-positive strains also harbored stx2, iha, saa, and lpfA(O113).

Distribution of virulence profiles related to new toxins and putative adhesins in Shiga toxin-producing Escherichia coli isolated from diverse sources in Brazil.

The distribution of virulence markers related to cytolethal distending toxin-V (CDT-V), subtilase cytotoxin (SubAB), the enterohemorrhagic Escherichia coli factor for adherence (Efa1), the adhesin similar to IrgA (Iha), the long polar fimbriae (LpfO113), the autoagglutinating adhesin (Saa), and the protein required for full expression of adherence of O157:H7 Sakai strain (ToxB) was investigated in 121 Shiga toxin-producing E. coli (STEC) strains isolated in Brazil. STEC strains were isolated from human infections (n=49), cattle (n=68) and ground meat samples (n=4). Overall, the lpfA(O113), iha, efa1, saa, and toxB sequences were observed in 89.2%, 87.6%, 47.1%, 43%, and 13.2% of the strains, respectively. The genes efa1 (96.6%) and toxB (27%) were only identified among eae-positive strains, while saa (83.8%), cdt-V (12.9%), and subAB (48.4%) just occurred in eae-negative STEC strains. STEC strains harboring cdt-V and subAB were for the first time described in the South American subcontinent. In addition, the simultaneous presence of cdt-V and subAB has not been previously reported, nor the presence of subAB in STEC O77, O79, O105, O174, and O178 serogroups. A diversity of virulence profiles was observed among the STEC strains studied. The most prevalent profile observed among eae-positive STEC strains mainly isolated from humans was eae efa1 iha lpfA(O113), whereas iha lpfA(O113) saa ehxA subAB prevailed among eae-negative STEC strains, mostly isolated from cattle and foods.