Contribution of wastewater-based epidemiology to SARS-CoV-2 screening in Brazil and the United States.

Wastewater-based epidemiology (WBE) is a valuable tool for investigating the existence, prevalence, and spread of pathogens, such as SARS-CoV-2, in a given population. WBE, proposed as part of the SARS-CoV-2 surveillance strategy for monitoring virus circulation, may complement clinical data and contribute to reducing the spread of the disease through early detection. In developing countries such as Brazil, where clinical data are scarce, information obtained from wastewater monitoring can be crucial for designing public health interventions. In the United States, the country with the largest number of confirmed SARS-CoV-2 cases worldwide, WBE programs have begun to be carried out to investigate correlations with coronavirus disease 2019 (COVID-19) clinical data and support health agencies in decision-making to prevent the spread of the disease. This systematic review aimed to assess the contribution of WBE to SARS-CoV-2 screening in Brazil and the United States and compare studies conducted in a developed and developing country. Studies in Brazil and the United States showed WBE to be an important epidemiological surveillance strategy in the context of the COVID-19 pandemic. WBE approaches are useful for early detection of COVID-19 outbreaks, estimation of clinical cases, and assessment of the effectiveness of vaccination program.

Diversity of bacteria carrying antibiotic resistance genes in hospital raw sewage in Southeastern Brazil.

In recent decades, antibiotic-resistant bacteria (ARB) emerged and spread among humans and animals worldwide. In this study, we evaluated the presence of ARB and antibiotic resistance genes (ARGs) in the raw sewage of two hospitals in Brazil. Sewage aliquots were inoculated in a selective medium with antibiotics. Bacterial identification was performed by MALDI-TOF and ARGs were assessed by polymerase chain reaction (PCR). A total of 208 strains from both hospitals were isolated (H1 = 117; H2 = 91). A wide variety of Enterobacterales and non-Enterobacterales species were isolated and most of them were Enterobacter spp. (13.0%), Proteus mirabilis (10.1%), and Klebsiella pneumoniae (9.6%). blaTEM and blaKPC were the most frequent ß-lactamase-encoding genes and the predominant macrolide resistance genes were mph(A) and mel. Many species had the three tetracycline resistance genes (tetD, tetM, tetA) and strB was the prevalent aminoglycoside resistance gene. Two Staphylococcus haemolyticus strains had the mecA gene. Quinolone, colistin, and vancomycin resistance genes were not found. This study showed that hospital raw sewage is a great ARB and ARG disseminator. Strict monitoring of hospital sewage treatment is needed to avoid the spread of these genes among bacteria in the environment.

Optimization of lipase production using fungal isolates from oily residues.

Lipases are triacylglycerol hydrolases that catalyze hydrolysis, esterification, interesterification, and transesterification reactions. These enzymes are targets of several industrial and biotech applications, such as catalysts, detergent production, food, biofuels, wastewater treatment, and others. Microbial enzymes are preferable for large scale production due to ease of production and extraction. Several studies have reported that lipases from filamentous fungi are predominantly extracellular and highly active. However, there are many factors that interfere with enzyme production (pH, temperature, medium composition, agitation, aeration, inducer type, and concentration, etc.), making control difficult and burdening the process. This work aimed to optimize the lipase production of four fungal isolates from oily residues (Penicillium sp., Aspergillus niger, Aspergillus sp., and Aspergillus sp.). The lipase-producing fungi isolates were morphologically characterized by optical and scanning electron microscopy. The optimal lipase production time curve was previously determined, and the response variable used was the amount of total protein in the medium after cultivation by submerged fermentation. A complete factorial design 32 was performed, evaluating the temperatures (28 °C, 32 °C, and 36 °C) and soybean oil inducer concentration (2%, 6%, and 10%). Each lipase-producing isolate reacted differently to the conditions tested, the Aspergillus sp. F18 reached maximum lipase production, compared to others, under conditions of 32 °C and 2% of oil with a yield of 11,007 (µg mL-1). Penicillium sp. F04 achieved better results at 36 °C and 6% oil, although for Aspergillus niger F16 was at 36 °C and 10% oil and Aspergillus sp. F21 at 32 °C and 2% oil. These results show that microorganisms isolated from oily residues derived from environmental sanitation can be a promising alternative for the large-scale production of lipases.

High prevalence of enteric viruses associated with acute gastroenteritis in pediatric patients in a low-income area in Vitória, Southeastern Brazil.

Acute gastroenteritis (AGE) is a significant cause of child mortality worldwide. In Brazil, despite the reduction in infant mortality achieved in recent years, many children still die because of undiagnosed AGE. The prevalence, viral load, and circulating genotypes of rotavirus A (RVA), human adenovirus (HAdV), and norovirus GII (NoV GII) were investigated in children with AGE during 12 months in Vitoria, Espírito Santo, Southeastern Brazil. Enteric viruses were detected in stool samples, quantified by quantitative polymerase chain reaction, sequenced, and compared phylogenetically. The overall prevalence was 93.3% (125/134). Cases of single infection (41.8%) and mixed infection (51.5%) were observed; in 21.6% of cases, all the three viruses were detected. RVA had the highest number of copies in all infections. Phylogenetic analysis revealed predominantly the presence of RVA genotype G3, followed by G2 and G9. HAdV clustered within subgroup C, but some samples harbored subgroups A, D, or F. All sequenced NoV-positive samples clustered within the prevalent genotype GII.4. The high prevalence of RVA, HAdV, and NoV in diarrheal feces clarifies the etiology of AGE in this population, and the presence of RVA in vaccinated children reinforces the importance of monitoring programs to identify the causes of gastroenteritis and contribute to the reliability of diagnosis.

Methylene blue removal using a nanomagnetic support: a response surface approach.

Textile wastewater is commonly released into water bodies without appropriated treatment, resulting in environmental damages. Processes involving separation and adsorption using nanomagnetic supports have been considered a promising alternative to address this concern. However, challenges concerning the low stability of nanoparticles and the reproducibility of experiments make their large-scale application difficult. In this study, we evaluated the efficiency of methylene blue (MB) removal by Fe3O4 nanoparticles coated with sodium dodecyl sulfate (MNP-SDS). The nanomaterial was characterized by transmission electron microscopy (TEM), X-ray diffraction (XRD), Raman spectroscopy, and Fourier-transform infrared spectroscopy (FTIR). The adsorption process was optimized in two stages using factorial design. In the first stage the most influential variables (reaction time, temperature, agitation, pH, and dye concentration) were selected based on the existing literature and applied in a fractional factorial (2(5-1)). In the second stage, the main variables identified were used in a complete factorial (32). The highest removal percentage was obtained using 15 g L-1 of MNP-SDS, which led to a qexp of 1.5 mg g-1. Isothermal analysis parameters and a negative Gibbs free energy indicate that the process was spontaneous, favorable, and that the data were best fitted to the Langmuir model.

Assessment of water and seafood microbiology quality in a mangrove region in Vitória, Brazil.

Mangroves are vital part of the local economy for some communities in the region of Vitória, Brazil. Oysters, mussels, and crabs, which are naturally abundant in the mangroves, are harvested and largely consumed in restaurants and by the population. In recent years, unusually high rates of annual gastroenteritis cases have been reported in the region suggesting an association between the consumption of contaminated shellfish and the development of gastrointestinal diseases. The objective of this study was to evaluate water samples and mussels collected in the mangrove region using bacterial indicator Escherichia coli and enteric viruses (adenovirus, rotavirus, and norovirus). Our results showed that the region of study is impacted by a continuous discharge of domestic sewage. Although E. coli was detected at low densities in water samples, mussels were shown to be 400 times more contaminated throughout the period of the study. Adenovirus and rotavirus genomes were detected by nested-polymerase chain reaction respectively in 76 and 88% of water samples and 100% of mussel samples. Norovirus was found in 4.8% of water samples and was not detected in the mussels. The screening of bivalves for the presence of health-significant enteric viruses can help in the prevention of outbreaks among shellfish consumers and contribute to improvement of the estuarine environment.

Detecting SARS-CoV-2 in sludge samples: A systematic review.

AIMS: This paper aims to review the main sludge concentration methods used for SARS-CoV-2 detection in sewage sludge samples, discussing the main methods and sample volume related to increased viral load. In addition, we aim to evaluate the countries associated with increased positivity rates for SARS-CoV-2 in sludge samples. METHODS: This systematic methodology was registered in PROSPERO and followed the PRISMA guidelines. The search was carried out in the SciELO, PubMed/MEDLINE, Lilacs, and Google Scholar databases in January-March 2022. Quantitative studies with conclusive results were included in this review. Concentration methods (polyethylene glycol (PEG), PEG + NaCl, gravity thickening, skimmed milk flocculation, ultrafiltration, filtration using charged filters, primary sedimentation, and anaerobic digestion), as well as detection methods (RTqPCR and reverse transcription droplet digital PCR assay) were evaluated in this review. The SPSS v23 software program was used for statistical analysis. RESULTS: PEG (with or without NaCl addition) and gravity thickening were the most used sludge concentration methods to detect SARS-CoV-2. The main method associated with increased viral load (>2,02 × 10^4 copies/mL) was PEG + NaCl (p < 0.05, Mann-Whitney test). The average positivity rate for SARS-CoV-2 in sludge samples was 61 %, and a correlation was found between the sludge volume and the viral load (ro 0.559, p = 0.03, Spearman correlation). CONCLUSION: The sludge volume may influence the SARS-CoV-2 load since the virus can adhere to solid particles in these samples. Other factors may be associated with SARS-CoV-2 load, including the methods used; especially PEG + NaCl may result in a high viral load detected in sludge, and may provide a suitable pH for SARS-CoV-2 recovery.

Impact of immobilization strategies on the activity and recyclability of lipases in nanomagnetic supports.

The use of enzymes immobilized on nanomagnetic supports has produced surprising results in catalysis, mainly due to the increase in surface area and the potential for recovery and reuse. However, the meticulous control of the process and difficulties in reproducibility have made industrial-scale applications unfeasible. Furthermore, the role of conjugation strategies in the catalytic activity and recycling of catalysts is unclear. Therefore, the objective of this study was to compare the conjugation of enzymes on nanomagnetic supports through physical adsorption (naked) or covalent bonding with mercaptopropyltrimethoxysilane (MPTS) and aminopropyltriethoxysilane (APTS) ligands. The free lipase obtained from Rhizomucor miehei was used as a model enzyme. Total protein and enzyme activity were determined using spectrophotometry (UV-Vis) and the p-nitrophenyl palmitate (p-NPP) hydrolysis method. The results indicated that a more significant enzyme surface loading does not always mean better immobilization success. The physical adsorption binding strategy had higher surface loading and low catalytic activity. On the other hand, covalent coupling with free NH2 had an excellent catalytic activity with very low surface loading. Finally, we show that recyclability can be improved with conjugation mediated by disulfide bonds. The findings presented here are essential for developing nanoconjugates with high enzymatic activity, which can guarantee the success of several industrial applications.

Nanomagnetic approach applied to microalgae biomass harvesting: advances, gaps, and perspectives.

Microalgae biomass is a versatile option for a myriad of purposes, as it does not require farmable land for cultivation and due of its high CO2 fixation efficiency during growth. However, biomass harvesting is considered a bottleneck in the process because of its high cost. Magnetic harvesting is a promising method on account of its low cost, high harvesting speed, and efficiency, which can be used to improve the results of other harvesting methods. Here, we present the state of the art of the magnetic harvesting method. Detailed approaches involving different nanomaterials are described, including types, route of synthesis, and functionalization, variables that interfere with harvesting, and recycling methods of nanoparticles and medium. In addition to discussing the overall perspectives of the method, we provide a guideline for future research.

Indigenous microalgae biomass cultivation in continuous reactor with anaerobic effluent: effect of dilution rate on productivity, nutrient removal and bioindicators.

Effluents from municipal wastewater treatment have been long recognized as suitable media for the cultivation of microalgae biomass. However, few studies report data concerning biomass productivity in continuous reactors using unsterilized wastewater effluents. This study focuses on indigenous microalgae strains that grow with native bacteria and are applicable for biomass production and tertiary wastewater treatment in continuous growth mode. Initially, five Chlorophyta strains were isolated and grown in batch mode to single out a potential inoculum for the experiments in continuous growth mode. The isolate Chlorella sp. L06 was selected and evaluated based on five dilution rates from 0.1 to 0.5 day-1 on continuous growth reactor using unsterilized secondary effluent as culture medium. Maximal volumetric biomass productivity of 283 mg L-1 day-1 was achieved at 0.3 day-1 without CO2 addition or air bubbling. Carbohydrates were the major fraction of the dried biomass, followed by proteins and then lipids. The highest removal rates of total nitrogen and phosphorus from the liquid phase were 13.0 and 1.4 mg L-1 day-1, respectively, and were obtained at 0.4 day-1. The maximal decay rate for E. coli (2.9 day-1) was achieved both at 0.3 and 0.4 day-1. Conclusively, Chlorella sp. L06 cultivation in unsterilized secondary effluent can be adjusted depending on the objective: for biomass production, a dilution rate of approximately 0.3 day-1 is recommended; and for tertiary treatment a rate of 0.4 day-1 is suggested.

Genetic diversity of indigenous common bean (Phaseolus vulgaris L.) rhizobia from the state of Minas Gerais, Brazil.

We characterized indigenous common bean rhizobia from five districts of the state of Minas Gerais, Brazil. The isolates were trapped by two common bean varieties, the Mineiro Precoce (Andean origin) and Ouro Negro (Mesoamerican origin). Analysis by BOX-PCR of selected isolates detected a high level of genetic diversity.

Surveillance of Enteric Viruses and Thermotolerant Coliforms in Surface Water and Bivalves from a Mangrove Estuary in Southeastern Brazil.

This study was conducted to evaluate the microbiological quality of a mangrove estuary in the Vitória Bay region, Espírito Santo, Brazil. We analyzed the presence and concentration of enteric viruses and thermotolerant coliforms in water, mussels (Mytella charruana and Mytella guyanensis), and oysters (Crassostrea rhizophorae), collected over a 13-month period. Human adenovirus, rotavirus A (RVA), and norovirus genogroup II were analyzed by quantitative PCR. The highest viral load was found in RVA-positive samples with a concentration of 3.0 × 104 genome copies (GC) L-1 in water samples and 1.3 × 105 GC g-1 in bivalves. RVA was the most prevalent virus in all matrices. Thermotolerant coliforms were quantified as colony-forming units (CFU) by the membrane filtration method. The concentration of these bacteria in water was in accordance with the Brazilian standard for recreational waters (< 250 CFU 100 mL-1) during most of the monitoring period (12 out of 13 months). However, thermotolerant coliform concentrations of 3.0, 3.1, and 2.6 log CFU 100 g-1 were detected in M. charruana, M. guyanensis, and C. rhizophorae, respectively. The presence of human-specific viruses in water and bivalves reflects the strong anthropogenic impact on the mangrove and serves as an early warning of waterborne and foodborne disease outbreaks resulting from the consumption of shellfish and the practice of water recreational activities in the region.

Harvesting Microalgal Biomass grown in Anaerobic Sewage Treatment Effluent by the Coagulation-Flocculation Method: Effect of pH

ABSTRACT Harvesting is a critical step in microalgal biomass production process for many reasons. Among the existing techniques available for harvesting and dewatering microalgal biomass, recovery from aqueous medium by coagulation-flocculation has been the most economically viable process, althoughit is highly dependent on pH. This study aims to assess alternative coagulants compared to the standard coagulant aluminum sulfate for microalgal biomass recovery from anaerobic effluent of domestic sewage treatment. The effluent quality was also analyzed after biomass recovery. Coagulants represented by modified tannin, cationic starch and aluminum sulfate recovered more than 90% of algae biomass, at concentrations greater than 80 mg/L, in the pH range 7-10. Cationic starch promoted higher microalgal biomass recovery with a wider pH range. Powdered seeds of Moringa oleifera and Hibiscus esculentus(okra) gum promoted biomass removal of 50%, only in the acidic range of pH. After sedimentation of the microalgal biomass, the effluents showed a removal of >80% for phosphorus and nitrogen values and >50% for BOD and COD when using aluminum sulfate, cationic starch and modified tannin as coagulants. Natural organic coagulants in a wide pH range can replace aluminum sulfate, a reference coagulant in microalgal biomass recovery, without decreasing microalgal biomass harvesting efficiency and the quality of the final effluent.

Isolamento e seleção de fungos produtores de lipases com base na atividade lipásica e no potencial hidrolítico sobre óleo comestível de soja e escuma de caixa de gordura / Isolation and selection of lipase-producing fungi based on lipase activity and hydrolytic potential on soybean oil and grease trap scum

RESUMO: O uso de biomassa fúngica como biocatalisadora lipásica representa uma atraente abordagem para o tratamento de águas residuais oleosas e produção de biodiesel, a partir de óleos e graxas residuais, devido à sua maior estabilidade, possibilidade de reuso e baixo custo. Neste trabalho foram obtidos cem isolados de fungos, a partir de escumas de caixa de gordura e esgoto, solo e tecidos necrosados de plantas e insetos, que foram avaliados quanto ao crescimento e à atividade lipásica, no meio de cultura básico, para atividade lipásica extracelular, e meio mineral mínimo + óleo de soja + rodamina, para atividade lipásica intracelular, com resposta positiva e diferenciada de 66 deles, inclusos como pertencentes aos gêneros Aspergillus , Beauveria , Botrytis , Cladosporium , Colletotrichum , Fusarium , Geotrichum , Penicillium , Rhizomucor e Verticillium . Na sequência, o potencial hidrolítico dos isolados Penicillium sp. F002 e Rhizomucor sp. F018 foi avaliado sobre óleo de soja comestível e escuma de caixa de gordura, em fermentação em estado sólido, através da quantificação das variáveis: produção de CO2, remoção do teor de óleos e graxas e crescimento da biomassa. Os resultados confirmaram a elevada atividade lipásica extracelular de Penicillium sp. F002 e a elevada atividade lipásica intracelular de Rhizomucor sp. F018. Portanto, o isolado Rhizomucor sp. F018 mostrou potencial para utilização em pesquisas futuras, na forma de células integrais lipásicas, para o tratamento de águas residuais oleosas e como biocatalisador na produção de biodiesel a partir de resíduos oleosos.

ABSTRACT: The use of fungal biomass as a lipase biocatalyst represents an attractive approach for the treatment of oil wastewater and production of biodiesel from oil and residual grease, due to its greater stability, possibility of reuse, and lower cost. In this work, a hundred filamentous fungi were isolated from grease trap and sewage scums, soil, and necrotized plants and insects tissues. The isolates were assessed for growth and lipase activity in the culture basic medium, for extracellular lipase activity, and mineral medium minimum + soybean oil + rhodamine, for intracellular lipase activity, with positive and differential response of 66 of them, including those belonging to the genera Aspergillus , Beauveria , Botrytis , Cladosporium , Colletotrichum , Fusarium , Geotrichum , Penicillium , Rhizomucor , and Verticillium . Following, previously selected Penicillium sp F002 and Rhizomucor sp. F018 isolates were evaluated in solid-state fermentation, for the hydrolytic potential on edible soybean oil and grease trap scum, quantified by: CO2 production, removal of the content of oils and greases, and biomass growth. Results confirmed the high extracellular lipase-activity of Penicillium sp. F002 and the high intracellular lipase activity of Rhizomucor sp. F018. Therefore, the isolated Rhizomucor sp. ECG18 showed potential for use in future research, in the form of whole-cell lipases, for oily wastewater treatment, and as a biocatalyst in the production of biodiesel from oil residues.

Genetic diversity of indigenous common bean (Phaseolus vulgaris L) rhizobia from the state of Minas Gerais, Brazil

We characterized indigenous common bean rhizobia from five districts of the state of Minas Gerais, Brazil. The isolates were trapped by two common bean varieties, the Mineiro Precoce (Andean origin) and Ouro Negro (Mesoamerican origin). Analysis by BOX-PCR of selected isolates detected a high level of genetic diversity.

Efeito de extratos de própolis verde sobre bactérias patogênicas isoladas do leite de vacas com mastite / Effect of green propolis extracts on patogenic bacteria isolated from milk of cows with mastitis

A sensibilidade, in vitro, de amostras de Staphylococcus aureus, Staphylococcus sp. coagulase negativos, Streptococcus agalactiae e bactérias do grupo dos coliformes, isoladas do leite de vacas com mastite, a diferentes extratos de própolis, na concentraçäo de 100 mg/ml, foi avaliada pela técnica do antibiograma em discos de papel de filtro com sobrecamada de meio de cultura. Os resultados mostraram que o extrato etanólico de própolis comercial, os extratos etanólico e, em menor proporçäo, o metanólico inibiram o crescimento das amostras de bactérias Gram-positivas, Staphylococcus aureus, Staphylococcus sp. coagulase negativos e Streptococcus agalactiae. Os extratos obtidos através da água, do acetato de etila e do clorofórmio näo inibiram nenhuma amostra bacteriana, assim como os veículos etanol e metanol puros utilizados como controle. A bactéria Gram-negativa testada, do tipo coliforme, näo apresentou sensibilidade a nenhum dos extratos. Verificaram-se diferenças significativas (p < 0,05) na sensibilidade aos extratos entre amostras bacterianas de uma mesma espécie, mas de origens diferentes. Nas amostras de Streptococcus agalactiae, os diâmetros dos halos de inibiçäo do crescimento bacteriano ao redor do disco foram maiores que aqueles observados para as amostras de Staphylococcus aureus e Staphylococcus sp. coagulase negativos. Todos estes resultados estimulam o prosseguimento de novas pesquisas sobre a utilizaçäo de extratos de própolis, em veículos adequados, com vistas ao tratamento da mastite bovina