Publications by authors named "Gislaine Fongaro"

32 Publications

The presence of SARS-CoV-2 RNA in human sewage in Santa Catarina, Brazil, November 2019.

Sci Total Environ 2021 Jul 8;778:146198. Epub 2021 Mar 8.

Microbiology Division, Faculty of Sciences, University of Burgos, Burgos, Spain. Electronic address:

Human sewage from Florianopolis (Santa Catarina, Brazil) was analyzed for severe acute respiratory syndrome coronavirus-2 (SARS-CoV2) from October 2019 until March 2020. Twenty five ml of sewage samples were clarified and viruses concentrated using a glycine buffer method coupled with polyethylene glycol precipitation, and viral RNA extracted using a commercial kit. SARS-CoV-2 RNA was detected by RT-qPCR using oligonucleotides targeting N1, S and two RdRp regions. The results of all positive samples were further confirmed by a different RT-qPCR system in an independent laboratory. S and RdRp amplicons were sequenced to confirm identity with SARS-CoV-2. Genome sequencing was performed using two strategies; a sequence-independent single-primer amplification (SISPA) approach, and by direct metagenomics using Illumina's NGS. SARS-CoV-2 RNA was detected on 27th November 2019 (5.49 ± 0.02 log SARS-CoV-2 genome copies (GC) L), detection being confirmed by an independent laboratory and genome sequencing analysis. The samples in the subsequent three events were positive by all RT-qPCR assays; these positive results were also confirmed by an independent laboratory. The average load was 5.83 ± 0.12 log SARS-CoV-2 GC L, ranging from 5.49 ± 0.02 log GC L (27th November 2019) to 6.68 ± 0.02 log GC L (4th March 2020). Our findings demonstrate that SARS-CoV-2 was likely circulating undetected in the community in Brazil since November 2019, earlier than the first reported case in the Americas (21st January 2020).
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http://dx.doi.org/10.1016/j.scitotenv.2021.146198DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7938741PMC
July 2021

Enteric viruses in lentic and lotic freshwater habitats from Brazil's Midwest and South regions in the Guarani Aquifer area.

Environ Sci Pollut Res Int 2021 Feb 20. Epub 2021 Feb 20.

Laboratory of Applied Virology, Department of Microbiology, Immunology, and Parasitology, Universidade Federal de Santa Catarina, João Pio Duarte Silva, 241, Córrego Grande, Florianópolis, SC, 88037-000, Brazil.

The present study reports the monitoring of viruses indicating fecal contamination in two distinct regions affected by poor management of wastewater located above the Guarani Aquifer, which is one of the biggest freshwater reservoirs in the world. In the city of Três Lagoas (located in the Midwest region, in the state of Mato Grosso do Sul), water samples were collected from Lagoa Maior, a lake used for recreation, and in Concórdia (located in the South region, in the state of Santa Catarina), from the Queimados River, which crosses the urban area. Four sampling sites were monitored from March to July 2018 in Lagoa Maior, and four sampling sites were monitored along the urban part of the Queimados River area over two periods (rainy and dry). Water samples were analyzed by concentration of Human adenovirus (HAdV), Norovirus (NoV), Rotavirus A (RAV), and Hepatitis A virus (HAV) for the Lagoa Maior samples and RVA, HAV, and Porcine circovirus 2 (PCV2) for the Queimados River samples. All sampling sites presented enteric viruses, demonstrating fecal input and potential contamination of groundwater. Results highlight the need for wastewater management to improve environmental health quality.
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http://dx.doi.org/10.1007/s11356-021-13029-yDOI Listing
February 2021

Swab pooling: A new method for large-scale RT-qPCR screening of SARS-CoV-2 avoiding sample dilution.

PLoS One 2021 4;16(2):e0246544. Epub 2021 Feb 4.

BiomeHub Biotechnologies, Florianópolis-SC, Brazil.

To minimize sample dilution effect on SARS-CoV-2 pool testing, we assessed analytical and diagnostic performance of a new methodology, namely swab pooling. In this method, swabs are pooled at the time of collection, as opposed to pooling of equal volumes from individually collected samples. Paired analysis of pooled and individual samples from 613 patients revealed 94 positive individuals. Having individual testing as reference, no false-positives or false-negatives were observed for swab pooling. In additional 18,922 patients screened with swab pooling (1,344 pools), mean Cq differences between individual and pool samples ranged from 0.1 (Cr.I. -0.98 to 1.17) to 2.09 (Cr.I. 1.24 to 2.94). Overall, 19,535 asymptomatic patients were screened using 4,400 RT-qPCR assays. This corresponds to an increase of 4.4 times in laboratory capacity and a reduction of 77% in required tests. Therefore, swab pooling represents a major alternative for reliable and large-scale screening of SARS-CoV-2 in low prevalence populations.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0246544PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7861376PMC
February 2021

Effect of dexamethasone as osteogenic supplementation in in vitro osteogenic differentiation of stem cells from human exfoliated deciduous teeth.

J Mater Sci Mater Med 2021 Jan 19;32(1). Epub 2021 Jan 19.

Center for Research on Dental Implants, Federal University of Santa Catarina, Delfino Conti Street, Florianópolis, 88040-900, Brazil.

In in vitro culture systems, dexamethasone (DEX) has been applied with ascorbic acid (ASC) and β-glycerophosphate (βGLY) as culture media supplementation to induce osteogenic differentiation of mesenchymal stem cells. However, there are some inconsistencies regarding the role of DEX as osteogenic media supplementation. Therefore, this study verified the influence of DEX culture media supplementation on the osteogenic differentiation, especially the capacity to mineralize the extracellular matrix of stem cells from human exfoliated deciduous teeth (SHED). Five groups were established: G1-SHED + Dulbecco's Modified Eagles' Medium (DMEM) + fetal bovine serum (FBS); G2-SHED + DMEM + FBS + DEX; G3-SHED + DMEM + FBS + ASC + βGLY; G4-SHED + DMEM + FBS + ASC + βGLY + DEX; G5-MC3T3-E1 + α Minimal Essential Medium (MEM) + FBS + ASC + βGLY. DNA content, alkaline phosphatase (ALP) activity, free calcium quantification in the extracellular medium, and extracellular matrix mineralization quantification through staining with von Kossa, alizarin red, and tetracycline were performed on days 7 and 21. Osteogenic media supplemented with ASC and β-GLY demonstrated similar effects on SHED in the presence or absence of DEX for DNA content (day 21) and capacity to mineralize the extracellular matrix according to alizarin red and tetracycline quantifications (day 21). In addition, the presence of DEX in the osteogenic medium promoted less ALP activity (day 7) and extracellular matrix mineralization according to the von Kossa assay (day 21), and more free calcium quantification at extracellular medium (day 21). In summary, the presence of DEX in the osteogenic media supplementation did not interfere with SHED commitment into mineral matrix depositor cells. We suggest that DEX may be omitted from culture media supplementation for SHED osteogenic differentiation in vitro studies.
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http://dx.doi.org/10.1007/s10856-020-06475-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7815568PMC
January 2021

Rural blackwater treatment by a full-scale Brazilian Biodigester Septic Tank: microbial indicators and pathogen removal efficiency.

Environ Sci Pollut Res Int 2021 May 13;28(18):23235-23242. Epub 2021 Jan 13.

Environmental Engineering Department, University of Oulu, 90570, Oulu, Finland.

The Brazilian Biodigester Septic Tank (BBST) is an on-site appropriate technology for blackwater treatment, which was developed to yield an effluent suitable for agricultural use. Although several studies have proven its efficacy for secondary blackwater treatment, there are few published studies about the microbiological quality of its effluent, and most of them focus on the quantification of total or thermotolerant coliforms. This study evaluates the performance of a BBST for the removal of human adenovirus (HAdV), Enterococcus spp., Salmonella sp., and Escherichia coli. The results further clarify the safety and risks associated with the reuse of the obtained effluent. The full-scale system consists of three 1.2 m interconnected reactors, with a blackwater input of 0.045 m/day, and hydraulic retention time of 80 days. Six sample campaigns were performed at different stages of the monthly operating cycle. The system presented an average removal efficiency of 5.09 log for E. coli, 3.22 log for Enterococcus spp., 1.2 log for Salmonella sp., and 3.0 log for HAdV. According to the World Health Organization standards, the obtained effluent is suitable for subsurface irrigation, and for use in crops that develop distant from the soil or highly mechanized crop systems.
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http://dx.doi.org/10.1007/s11356-020-12229-2DOI Listing
May 2021

Hepatitis E Virus in Manure and Its Removal by Biodigestion in Intensive Production Farms, Santa Catarina, Brazil, 2018-2019.

Microorganisms 2020 Dec 21;8(12). Epub 2020 Dec 21.

Laboratory of Applied Virology, Department of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina, Florianópolis, SC 88040-900, Brazil.

Hepatitis E virus (HEV) is an important enteric agent that can circulate in swine; it is excreted in manure, and of zoonotic interest. The present study investigated, by RT-qPCR, the circulation of HEV in swine manure from different types of pig farms (maternity, nursery, and grow-finish farms) in Santa Catarina State, the major pig production area of Brazil, and also evaluated the HEV removal efficiency of psychrophilic anaerobic biodigesters (PABs). While HEV was consistently detected in manure from grow-finish pig farms (>4 log HEV genome copies (GC) L), the virus was not detected in manure from maternity and nursery farms. These findings suggest a potential high biosafety status during primary-swine production, with a subsequent contamination in grow-finish production. The anaerobic biodigestion process reduced more than 2 log HEV GC in the processed swine manure. However, the virus concentration in final effluent remained high, with an average value of 3.85 log HEV GC L. Consequently, our results demonstrate that PABs can be a robust tool for effective inactivation of HEV, while reinforcing the need for sanitary surveillance and legislation of swine manure-derived biofertilizers, to avoid the spread of zoonotic enteric pathogens such as HEV.
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http://dx.doi.org/10.3390/microorganisms8122045DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7766788PMC
December 2020

Utilization of seawater and wastewater from shrimp production in the fermentation of papaya residues to ethanol.

Bioresour Technol 2021 Feb 7;321:124501. Epub 2020 Dec 7.

Laboratory of Microbiology and Bioprocesses, Federal University of Fronteira Sul, Erechim, Brazil. Electronic address:

Seawater (SW) and wastewater from shrimp production (WSP) were used as a solvent for the fermentation of papaya residues (Carica papaya) by Wickerhamomyces sp. UFFS-CE-3.1.2 and Saccharomyces cerevisiae CAT-1. For comparative purposes and evaluation of the effect of salinity, ultrapure water (UW) was used as control. Fermentative parameters were evaluated in Plackett-Burman planning to assess ethanol production's significant variables. Urea supplementation was the only variable not significant for the proposed process, suggesting that papaya residues contain all the nutrients needed for fermentation. The experiments conducted with the different water sources resulted in similar concentrations of ethanol. Maximum ethanol concentration was obtained after nine h of fermentation usingWickerhamomycessp. UFFS-CE-3.1.2 (27.31 ± 1.40 g L) and 12 h using S. cerevisiaeCAT-1 (24.53 ± 0.68 g L). This study demonstrated that SW and WSP could replace freshwater without affecting ethanol production. Papaya residues from the fruit and vegetable sectors can be considered a promising substrate source for ethanol production.
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http://dx.doi.org/10.1016/j.biortech.2020.124501DOI Listing
February 2021

One-step procedure for peroxidase concentration, dye separation, and color removal by aqueous two-phase system.

Environ Sci Pollut Res Int 2021 Feb 31;28(8):9097-9106. Epub 2020 Oct 31.

Laboratory of Microbiology and Bioprocess, Federal University of Fronteira Sul, Erechim, Brazil.

The aim of our study is to develop a one-step procedure to remove and degrade dyes from wastewater using a low-cost and efficient system based on aqueous two-phase system (ATPS), a well-known technique used to concentrate and recover enzymes. We investigated the catalytic proprieties of rice bran peroxidase (RBP) and found that this homemade enzyme can remain bound to its substrate for up to 5 days in controlled environments, without denaturing and while maintaining stable oxidation reduction potential (ORP) and pH. This biomolecule showed affinity for the ATPS technique prepared with polyethylene glycol and salt, which improved the relative activity up to 170%. The red dye separation in ATPS top phase was achieved in 3 min, in the RBP presence, with 100% of efficiency, and color removal of 87% was obtained in 24 h of enzymatic reaction. The process has promise to be scaled up to 10-fold and to reuse the reagents from the bottom phase of the ATPS.
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http://dx.doi.org/10.1007/s11356-020-11412-9DOI Listing
February 2021

Extremophile Microbial Communities and Enzymes for Bioenergetic Application Based on Multi-Omics Tools.

Curr Genomics 2020 May;21(4):240-252

1Department of Microbiology, Immunology, and Parasitology, Federal University of Santa Catarina, Florianópolis, SC, Brazil; 2Laboratory of Microbiology and Bioprocess, Federal University of Fronteira Sul, Erechim, RS, Brazil; 3Department of Chemical and Food Engineering, Federal University of Santa Catarina, Florianópolis, SC, Brazil; 4Department of Dentistry, Federal University of Santa Catarina, Florianópolis, SC, Brazil.

Genomic and proteomic advances in extremophile microorganism studies are increasingly demonstrating their ability to produce a variety of enzymes capable of converting biomass into bioenergy. Such microorganisms are found in environments with nutritional restrictions, anaerobic environments, high salinity, varying pH conditions and extreme natural environments such as hydrothermal vents, soda lakes, and Antarctic sediments. As extremophile microorganisms and their enzymes are found in widely disparate locations, they generate new possibilities and opportunities to explore biotechnological prospecting, including biofuels (biogas, hydrogen and ethanol) with an aim toward using multi-omics tools that shed light on biotechnological breakthroughs.
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http://dx.doi.org/10.2174/1389202921999200601144137DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7521039PMC
May 2020

A review on alternative bioprocesses for removal of emerging contaminants.

Bioprocess Biosyst Eng 2020 Dec 17;43(12):2117-2129. Epub 2020 Jul 17.

Laboratory of Microbiology and Bioprocess, Universidade Federal da Fronteira Sul, Erechim, Brazil.

Emerging contaminants (ECs) include endocrine-disrupting compounds, pharmaceuticals (lipid regulators, antibiotics, diuretics, non-steroid anti-inflammatory drugs, stimulant drugs, antiseptics, analgesic, beta blockers), detergents, disinfectants, and personal care products. The residues from these compounds have become a concerning because of their bioactive presence on environmental matrices, especially water bodies. The development of technologies, aiming the secure and efficient removal of these compounds from the environment or event to remove them before they achieve the environment, is necessary. In these context, the present review is about promising eco-friendly, low-cost and specially applied, including biological processes using microalgae, bacteria, enzymes produced by fungi, and adsorbent materials such as those recycled from other processes waste. The processes where revised considering the removal mechanism and the efficiency rate.
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http://dx.doi.org/10.1007/s00449-020-02410-9DOI Listing
December 2020

Nutritional, Energy and Sanitary Aspects of Swine Manure and Carcass Co-digestion.

Front Bioeng Biotechnol 2020 29;8:333. Epub 2020 Apr 29.

Laboratory of Applied Virology, Department of Microbiology, Parasitology and Immunology, Federal University of Santa Catarina (UFSC), Florianópolis, Brazil.

Renewable energy can assist the management of the effects of population growth and rapid economic development on the sustainability of animal husbandry. The primary aim of renewable energy is to minimize the use of fossil fuels via the creation of environmentally friendly energy products from depleted fossil fuels. Digesters that treat swine manure are extensively used in treatment systems; and inclusion of swine carcasses can increase the organic loading rate (OLR) thereby improving biogas yield and productivity on farms. However, the characteristics of the components including animal residues, proteins, lipids, remains of undigested feed items, antimicrobial drug residues, pathogenic microorganisms and nutrient contents, are complex and diverse. It is therefore necessary to manage the anaerobic process stability and digestate purification for subsequent use as fertilizer. Efficient methane recovery from residues rich in lipids is difficult because such residues are only slowly biodegradable. Pretreatment can promote solubilization of lipids and accelerate anaerobic digestion, and pretreatments can process the swine carcass before its introduction onto biodigesters. This review presents an overview of the anaerobic digestion of swine manure and carcasses. We analyze the characteristics of these residues, and we identify strategies to enhance biogas yield and process stability. We consider energy potential, co-digestion of swine manure and carcasses, physical, chemical, and biological pretreatment of biomass, sanitary aspects of swine manure and co-digestates and their recycling as fertilizers.
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http://dx.doi.org/10.3389/fbioe.2020.00333DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7200981PMC
April 2020

and sp. as Keratinase Producers Using Swine Hair From Agroindustrial Residues.

Front Bioeng Biotechnol 2020 11;8:71. Epub 2020 Feb 11.

Laboratory of Microbiology and Bioprocess, Federal University of Fronteira Sul, Erechim, Brazil.

Technological processes mediated by microorganisms and enzymes are promising alternatives for treatment of recalcitrant residues. Keratinases hydrolyze keratin, the primary component of some wastes generated in many industrial activities. The present study was designed to evaluate strategies for obtaining keratinases produced by fungi using submerged fermentation and two residues as substrates, chicken feathers and swine hair. Two fungi isolated from feather residues showed potential for keratinase production, and sp. These were subjected to submerged fermentation using chicken feathers and swine hair prepared in three conditions (microbial concentration reduction, sterilization and hydrogen peroxide). The residual mass was quantified and tested for keratinase production. The most potent enzymatic extract was used in the precipitation technique with salts and organic solvents. The best results of enzymatic activity were obtained using , on the 6thday of fermentation, obtaining 243.25 U mL using sterilized swine hair as the substrate. sp. showed the highest keratinolytic activity on the 9thday, 113.50 U mL using feathers as the substrate. The highest degradation percentage was 59.20% (w/w) in swine hair and the precipitation technique, with relative activities close to 50%. The results are promising for the application of residues and microorganisms in biotechnological processes of economic and environmental interest.
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http://dx.doi.org/10.3389/fbioe.2020.00071DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7026017PMC
February 2020

Occurrence of Hepatitis E Virus in Pigs and Pork Cuts and Organs at the Time of Slaughter, Spain, 2017.

Front Microbiol 2019 28;10:2990. Epub 2020 Jan 28.

Division of Microbiology, Department of Biotechnology and Food Science, Universidad de Burgos, Burgos, Spain.

Zoonotic hepatitis E, mainly caused by hepatitis E virus (HEV) genotype (gt) 3, is a foodborne disease that has emerged in Europe in recent decades. The main animal reservoir for genotype 3 is domestic pigs. Pig liver and liver derivates are considered the major risk products, and studies focused on the presence of HEV in pig muscles are scarce. The objective of the present study was to evaluate the presence of HEV in different organs and tissues of 45 apparently healthy pigs from nine Spanish slaughterhouses (50% national production) that could enter into the food supply chain. Anti-HEV antibodies were evaluated in serum by an ELISA test. Ten samples from each animal were analyzed for the presence of HEV RNA by reverse transcription real-time PCR (RT-qPCR). The overall seroprevalence obtained was 73.3% (33/45). From the 450 samples analyzed, a total of 26 RT-qPCR positive samples were identified in the liver (7/45), feces (6/45), kidney (5/45), heart (4/45), serum (3/45), and diaphragm (1/45). This is the first report on detection of HEV RNA in kidney and heart samples of naturally infected pigs. HEV RNA detection was negative for rib, bacon, lean ham, and loin samples. These findings indicate that pig meat could be considered as a low risk material for foodborne HEV infection.
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http://dx.doi.org/10.3389/fmicb.2019.02990DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6997137PMC
January 2020

Co-contamination of food products from family farms in an environmental disaster area in Southeast Brazil with pathogenic bacteria and enteric viruses.

Arch Virol 2020 Mar 23;165(3):715-718. Epub 2019 Dec 23.

Applied Virology Laboratory, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil.

In the present study, we evaluated the degree of contamination of fresh vegetables, cheeses and jellies from disaster area in Brazil with bacteria and enteric viruses. Food samples (n = 350) were tested for Escherichia coli, Salmonella spp., Listeria monocytogenes, Staphylococcus spp., and enteric viruses (rotavirus A (RVA), human adenovirus (HAdV), hepatitis A virus (HAV), and human norovirus (HNoV). E. coli was present in 56% of the samples, Salmonella spp. was present in 14% of the samples, L. monocytogenes and Staphylococcus spp. (coagulase-positive) were present in 36% of the samples. The enteric viruses RVA and HAdV were detected in cheeses and vegetables.
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http://dx.doi.org/10.1007/s00705-019-04501-9DOI Listing
March 2020

Advanced oxidation processes applied for color removal of textile effluent using a home-made peroxidase from rice bran.

Bioprocess Biosyst Eng 2020 Feb 1;43(2):261-272. Epub 2019 Oct 1.

Laboratory of Microbiology and Bioprocess, Federal University of Fronteira Sul, Erechim, Brazil.

Enzymes are becoming tools in industrial processes because of several advantages, including activity in mild environmental conditions, and high specificity. Peroxidase, for one, stably oxidizes several substrates. The present study aimed to develop advanced oxidation processes (AOP), using non-commercial rice bran peroxidase to remove color and toxicity of synthetic textile wastewater. Using a microwave and shaker system, we obtained 38.9% and 100% of effluent color removal after peroxidase treatment, respectively. In addition, the shaker system decants residual dye particles through filtration, providing the textile industry with an economical and environmentally viable alternative to effluent treatment. In toxicity tests results, both treatment systems damaged the used genetic material. This damage occurs because of industrial discharge of wastewater into water bodies; effluent dilution reduced this damage. The data suggest that peroxidase as a textile effluent treatment has potential uses in industrial processes, because rice bran peroxidase has demonstrated affinity with dyes.
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http://dx.doi.org/10.1007/s00449-019-02222-6DOI Listing
February 2020

Removal of chromium from wastewater by swine hair residues applied as a putative biofilter.

Environ Sci Pollut Res Int 2019 Nov 11;26(32):33014-33022. Epub 2019 Sep 11.

Laboratory of Microbiology and Bioprocess, University of Fronteira Sul, Erechim, Brazil.

Swine production chain generates residues with potential application in environmental processes. This study aimed at the use of swine hair as a potential biofilter for hexavalent chromium (Cr(VI)) removal from wastewater of tannery industry. The hair was pretreated using HO in alkaline medium, and statistical analysis was carried out to evaluate the hair degradation, as well the Cr(VI) removal by the potential pretreated biofilter. The results showed 99% of Cr(VI) removal in 105 min of treatment in large pH range (1-10). Treated and untreated effluents were submitted to cytotoxicity study using vegetable and animal cells, demonstrating a significant reduction on toxicity to both cells. Therefore, swine hair demonstrated to be a promising residue for heavy metal removal on the perspective of an environmentally friendly technique.
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http://dx.doi.org/10.1007/s11356-019-06313-5DOI Listing
November 2019

New perspectives for weeds control using autochthonous fungi with selective bioherbicide potential.

Heliyon 2019 May 11;5(5):e01676. Epub 2019 May 11.

Laboratory of Agroecology, Federal University of Fronteira Sul, Erechim campus, RS-135, Rural Area, Erechim, Brazil.

The prospection of bioherbicides has been an alternative to weed control, aiming at mitigating chemical risks to human, animal and environmental health due to extreme use of synthetic herbicides. In the present study, various fungi were isolated from plants with symptoms of fungal diseases for bioherbicide purposes against weeds (, and ). Fungi isolated were identified by molecular methods and enzymatic products obtained by fungi fermentation (cellulase, lipase, peroxidase, and amylase) were quantified. Bioherbicide selectivity study was performed on crops (soybean and corn), as well as on resistant weeds. Among the isolated fungi, , and presented bioherbicide potential. , in particular, presented the highest effect on (popular name - Mexican fire plant), causing up to 60% of foliar damage, without presenting phytotoxicity against corn crop. New perspectives for weeds control and their use in corn crops were prospected, considering the bioherbicide selectivity described in this study.
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http://dx.doi.org/10.1016/j.heliyon.2019.e01676DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6517331PMC
May 2019

Evaluation of deammonification reactor performance and microrganisms community during treatment of digestate from swine sludge CSTR biodigester.

J Environ Manage 2019 Sep 4;246:19-26. Epub 2019 Jun 4.

Western Paraná State University, 85819-110, Cascavel, PR, Brazil; Embrapa Suínos e Aves, 89715-899, Concórdia, SC, Brazil; Federal University of Fronteira Sul, 99700-000, Erechim, RS, Brazil. Electronic address:

Digestate from anaerobic processes still contains relatively high amount of total organic carbon (TOC) that can inhibit deammonification. In this sense, the present study investigated the interference of TOC in a lab-scale expanded granular sludge bed (EGSB) deammonification reactor treating digestate from a continuous stirred tank reactor (CSTR) swine sludge biodigester. Additionally, the microorganisms community was analyzed when the process was submitted to different operational conditions. The study was divided into three phases according to the C/N ratio (0, 0.5 and 1 for phase I, phase II and phase III, respectively). At phase I the average nitrogen removal efficiency (NRE) was 65 ± 1.6%. With the increase of TOC in phase II (156 ± 8.15 mg L) the average NRE was 61 ± 9.8% which is statically equivalent to phase I (p < 0.05). On the other hand, at phase III (TOC was increased to 255 ± 3.50 mg L) the NRE decreased to 50 ± 3.9% which was 22% lower than in phase II. Stoichiometric coefficients of N was close to theoretical values during all experimental phases, while stoichiometric coefficient of N-NO was lower than theoretical values specially during phase III. Ca. Jettenia was favored when the reactor was fed with digestate although its proportion decreased in phase III. Thus, at the conditions employed in the present study it is recommended to use a C/N ratio of 0.5 (TOC concentration around 156 mg L) to treat digestate by deammonification process, in order to not diminish anammox microorganisms abundance. Thereby, the microorganisms community can be modulated based on carbon and nitrogen loading rates of a deammonification reactor for swine manure treatment purpose.
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http://dx.doi.org/10.1016/j.jenvman.2019.05.113DOI Listing
September 2019

Household-based biodigesters promote reduction of enteric virus and bacteria in vulnerable and poverty rural area.

Environ Pollut 2019 Sep 21;252(Pt A):8-13. Epub 2019 May 21.

Laboratory of Applied Virology, Universidade Federal de Santa Catarina, Florianópolis, SC, Brazil. Electronic address:

The present study evaluated the river water quality improvement by implementation of household-based biodigesters in vulnerability and poverty rural area, in Minas Gerais State-Brazil. For that, 78 household-based biodigesters were installed for domestic wastewater treatment. Wastewater was collected before and after treatment and the physicochemical parameters and pathogens removal (human adenovirus (HAdV), hepatitis A (HAV) virus, Salmonella sp. and Escherichia coli) were evaluated; Additionally, river water was sampled before and after the household-based biodigesters implementation, to verify the contamination reduction and the positive impact of domestic wastewater treatment on waterborne pathogen reduction, considering HAdV, HAV, Salmonella sp. and E. coli quantification. The applicability in real-scale of decentralized treatment systems using household-based biodigesters promoted reduction of 90, 99, 99.99 and 99.999% from HAV, Salmonella sp., E. coli and HAdV from domestic wastewater, respectively; The river water quality improvement before the wastewater treatment application was highlight in the present study, considering that the reduction of waterborne pathogens in this water in 90, 99.99 and 99.999% of E. coli, HAV and HAdV, respectively (Salmonella sp. was not detected in river water). In general, this is an important study for encouraging the decentralized sanitation in vulnerable and poverty area, as well in rural sites, considering the positive impact of this implementation on public health.
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http://dx.doi.org/10.1016/j.envpol.2019.05.104DOI Listing
September 2019

The Relationship Between Human Adenovirus and Metals and Semimetals in the Waters of the Rio Doce, Brazil.

Arch Environ Contam Toxicol 2019 Jul 11;77(1):144-153. Epub 2019 Apr 11.

Department of Civil Engineering, Escola de Minas, Federal University of Ouro Preto, Campus Universitário Morro do Cruzeiro, s/n, Ouro Preto, MG, 35400-000, Brazil.

The 2015 rupture of the Fundão dam near the district of Bento Rodrigues in Mariana, Minas Gerais, Brazil, released around 50 million m of iron ore tailings. The first tributary of the Rio Doceto receive this waste was the Gualaxo do Norte River. Many groups in Brazil and from around the world have studied the environmental and social impacts of this disaster. However, relationships between the introduction of mining waste, the geological complexity of the area, and the presence of pathogenic organisms have not yet been investigated. The present study aimed to measure the concentrations of enteric pathogens along the Gualaxo do Norte River after the environmental disaster and to correlate their abundance with the presence of metals and semimetals coming from both mining tailings and geological sources. For this purpose, we collected water samples from 27 stations along the entire basin during a hydrological year. The concentrations of metals and semimetals measured in this study were generally within limits established by national and international legislation, except for those of iron and manganese. Positive correlations between the human adenovirus (HAdV) and arsenic, barium, iron, lead, manganese, and nickel were confirmed, allowing us to observe that there is an abundance of the potentially infectious virus present in the studied sites containing metal/semimetal concentrations. These studies indicate the importance of investigations that consider viral enteric pathogens complexed with metals and may favor the stability and prolongation of the infectivity of such pathogens in water destined for human and animal usage.
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http://dx.doi.org/10.1007/s00244-019-00625-wDOI Listing
July 2019

Mineral Waste Containing High Levels of Iron from an Environmental Disaster (Bento Rodrigues, Mariana, Brazil) is Associated with Higher Titers of Enteric Viruses.

Food Environ Virol 2019 06 12;11(2):178-183. Epub 2019 Feb 12.

Microbiology Section, Department of Biotechnology and Food Science, Universidad de Burgos, Plaza Misael Bañuelos s/n, 09001, Burgos, Spain.

Although the effects of heavy metals on the behavior, including infectivity, of bacteria have been studied, little information is available about their effects on enteric viruses. We report an investigation of effects on the biosynthesis of human adenoviruses (HAdV) and hepatitis A (HAV) of waters contaminated with mineral waste following an environmental disaster in Mariana City, Minas Gerais State, Brazil. The study area was affected on November 5, 2015, by 60 million m of mud (containing very high concentrations of iron salts) from a mining reservoir (Fundão), reaching the Gualaxo do Norte River (sites evaluated in this study), the "Rio Doce" River and finally the Atlantic Ocean. We found substantial counts of infectious HAdV and HAV (by qPCR) in all sampled sites from Gualaxo do Norte River, indicating poor basic sanitation in this area. The effects of iron on viral infection processes were evaluated using HAdV-2 and HAV-175, as DNA and RNA enteric virus models, respectively, propagated in the laboratory and exposed to this contaminated water. Experiments in field and laboratory scales found that the numbers of plaque forming units (PFU) of HAdV and HAV were significantly higher in contaminated water with high iron concentrations than in waters with low iron concentration (< 20 µg/L of iron). These findings indicate that iron can potentiate enteric virus infectivity, posing a potential risk to human and animal health, particularly during pollution disasters such as that described here in Mariana, Brazil.
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http://dx.doi.org/10.1007/s12560-019-09373-5DOI Listing
June 2019

Cellulolytic enzyme production from agricultural residues for biofuel purpose on circular economy approach.

Bioprocess Biosyst Eng 2019 May 19;42(5):677-685. Epub 2019 Jan 19.

Laboratory of Microbiology and Bioprocess, Federal University of Fronteira Sul, ERS 135-Km 72, 200, Erechim, 99700-000, Brazil.

This study evaluated the production of cellulolytic enzymes from different agricultural residues. The crude enzyme extract produced was characterized and applied for saccharification of some agricultural residues. Maximum cellulolytic activities were obtained using soybean hulls. All enzymatic activities were highly stable at 40 °C at a pH range of 4.5-5.5. For stability at low temperatures, the enzyme extract was stored at freezing temperature and cooling for about 290 days without major loss of activity. The K values found for total cellulase (FPase), endoglucanase (CMCase), and xylanase were 19.73 mg ml, 0.65 mg ml, and 22.64 mg ml, respectively, and V values were 0.82 mol min mg, 0.62 mol min mg, and 104.17 mol min mg to cellulose, carboxymethyl cellulose, and xylan, respectively. In the saccharification tests, the total amount of total reducing sugars (TRS) released from 1 g of soybean hulls catalyzed by the enzymes present in the crude enzyme extract was 0.16 g g dry substrate.
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http://dx.doi.org/10.1007/s00449-019-02072-2DOI Listing
May 2019

Sanitary effectiveness and biogas yield by anaerobic co-digestion of swine carcasses and manure.

Environ Technol 2020 Feb 21;41(6):682-690. Epub 2018 Aug 21.

Western Paraná State University - UNIOESTE/CCET/PGEAGRI, Cascavel, Brazil.

The present study evaluated anaerobic co-digestion of swine manure and swine carcasses for biogas yield and inactivation/behaviour of pathogens purpose. Biochemical Methane Production tests were performed with samples containing ratios of 3, 7.5 and 15 kg m. For pathogens inactivation experiments known amounts of model microrganisms (sensitive and resistant) were artificially inoculated in anaerobic reactors at 24°C and 37°C. The addition of carcass resulted in an increase until 119% of biogas yield compared to swine manure mono-digestion. , and PCV2 were reduced >3log (24°C or 37°C) during 30 days. At 37°C, MS2 and PhiX-174 were reduced 3log and 1.8log respectively. At 24°C, MS2 reduced 1.5 log and PhiX-174 did not present any decay over 30 days. Considering the most resistant biomarkers pathogens, as bacteriophage, we recommend the swine carcasses pre-treatment, such as high temperatures, for sanitary security.
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http://dx.doi.org/10.1080/09593330.2018.1508256DOI Listing
February 2020

Effect of pretreatments on corn stalk chemical properties for biogas production purposes.

Bioresour Technol 2018 Oct 21;266:116-124. Epub 2018 Jun 21.

Federal University of Fronteira Sul (UFFS), Laboratory of Microbiology and Bioprocesses, RS 135, Km 72, 99700-000 Erechim, RS, Brazil. Electronic address:

Different pretreatments were evaluated on corn stalk (Zea mays) applied as a lignocellulosic source in anaerobic co-digestion with swine manure, using sulfuric acid (HSO) and hydrogen peroxide (HO) for biogas production purposes. Using HSO we achieved a 75.1% removal of the hemicellulose fraction, in low acid concentrations (0.75% v.v). However, this technique inhibited the co-digestion process. Pretreatment with 12% of HO (pH 11.5) increased the cellulose fraction by 73.4% and reduced the lignin content by 71.6%. This pretreatment is recommended for biogas production, as it increased the final volume of biogas by 22% and reduced the digestion time by one third. So, a promising alternative was obtained in order to facilitate the anaerobic digestion of the carbohydrates present in this biomass.
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http://dx.doi.org/10.1016/j.biortech.2018.06.069DOI Listing
October 2018

Genotypic characterization and assessment of infectivity of human waterborne pathogens recovered from oysters and estuarine waters in Brazil.

Water Res 2018 06 9;137:273-280. Epub 2018 Mar 9.

Universidade Estadual de Campinas, Instituto de Biologia, Departamento de Biologia Animal, Laboratório de Protozoologia, Campinas, São Paulo CEP 13083-970, Brazil.

Waterborne, food-borne and sewage-borne pathogens are a major global concern, with the annual recurrence, most notably during the summer, of outbreaks of gastroenteritis of unconfirmed etiology associated with recreational activities in marine environments. The consumption of contaminated water-based foodstuffs is also related to outbreaks of human illness. The main goals of the present study were: i) to identify the genetic assemblages of Giardia duodenalis cysts in growing and depurated oysters destined for human consumption on the southern coast of São Paulo, Brazil; ii) to verify the main circulating G. duodenalis assemblages and their subtypes in different brackish waters used for the production of mollusks and for recreational purposes; iii) to track the contamination of growing and depurated oysters by the human adenovirus and identify the infectivity of adenoviral particles recovered from oysters before and after depuration; iv) to evaluate the occurrence and genotype of the free-living amoebae of the genus Acanthamoeba in brackish water and oysters from all the sites described above. Four sampling sites in the Cananeia estuary were selected to search for pathogenic and amphizoic protozoa (Giardia and Acanthamoeba respectively): site 1: oyster growth, site 2: catchment water (before UV depuration procedure), site 3: filter backwash (filtration stage of water treatment) and site 4: oyster depuration tank. Oysters at sites 1 and 4 were evaluated for the presence of adenovirus (HAdV). Analysis consisted of conventional microbiological as well as molecular methods. Giardia duodenalis were detected in all the water sites analyzed and the molecular analysis revealed that sub-assemblage AII was the most frequently distributed throughout the estuarine environment, although one sample was identified as belonging to the assemblage C. Acanthamoeba were also isolated from different locations of the estuarine area, and were detected at all the analyzed sites. The majority of isolates belonged to the T3 genotype, while the T4 genotype was identified once. The sequencing reaction of Giardia duodenalis revealed the contamination of three batches of depurated oysters by the sub-assemblage AII. With respect to viruses, seven batches of oysters (four growing and three depurated) were found to be harboring infectious HAdV particles when submitted to plaque assay. Overall, the results of the sequencing reactions combined with the plaque assay revealed that the isolates of Giardia duodenalis and the infectious HAdV particles identified in oyster tissues have the potential to infect humans and pose a threat if consumed raw or lightly cooked. This is the first report on the sub-assemblage AII identified in oysters which are submitted to a cleaning and disinfection procedure prior to human consumption in Brazil. Acanthamoeba specific genotypes were also identified for the first time in a recreational estuarine area in Brazil, contributing to knowledge of their molecular and environmental epidemiology, which is considered scarce even in marine and estuarine areas of the world.
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http://dx.doi.org/10.1016/j.watres.2018.03.024DOI Listing
June 2018

Different Behavior of Enteric Bacteria and Viruses in Clay and Sandy Soils after Biofertilization with Swine Digestate.

Front Microbiol 2017 31;8:74. Epub 2017 Jan 31.

Microbiology Section, Department of Biotechnology and Food Science, Faculty of Sciences, Universidad de Burgos Burgos, Spain.

Enteric pathogens from biofertilizer can accumulate in the soil, subsequently contaminating water and crops. We evaluated the survival, percolation and leaching of model enteric pathogens in clay and sandy soils after biofertilization with swine digestate: PhiX-174, mengovirus (vMC), Typhimurium and O157:H7 were used as biomarkers. The survival of vMC and PhiX-174 in clay soil was significantly lower than in sandy soil (T values of 10.520 ± 0.600 vs. 21.270 ± 1.100 and 12.040 ± 0.010 vs. 43.470 ± 1.300, respectively) and PhiX-174 showed faster percolation and leaching in sandy soil than clay soil (T values of 0.46 and 2.43, respectively). . Typhimurium was percolated and inactivated more slowly than O157:H7 (T values of 9.340 ± 0.200 vs. 6.620 ± 0.500 and 11.900 ± 0.900 vs. 10.750 ± 0.900 in clay and sandy soils, respectively), such that O157:H7 was transferred more quickly to the deeper layers of both soils evaluated (percolation). Our findings suggest that O157:H7 may serve as a useful microbial biomarker of depth contamination and leaching in clay and sandy soil and that bacteriophage could be used as an indicator of enteric pathogen persistence. Our study contributes to development of predictive models for enteric pathogen behavior in soils, and for potential water and food contamination associated with biofertilization, useful for risk management and mitigation in swine digestate recycling.
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http://dx.doi.org/10.3389/fmicb.2017.00074DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5281563PMC
January 2017

Propidium Monoazide Integrated with qPCR Enables the Detection and Enumeration of Infectious Enteric RNA and DNA Viruses in Clam and Fermented Sausages.

Front Microbiol 2016 15;7:2008. Epub 2016 Dec 15.

Division of Microbiology, Department of Biotechnology and Food Science, Universidad de Burgos Burgos, Spain.

The increase of foodborne viral outbreaks highlights the need for a rapid and sensitive method for the prediction of viral infectivity in food samples. This study assesses the use of propidium monoazide (PMA) coupled with real-time PCR methods (RT-qPCR or qPCR for RNA or DNA viruses, respectively) in the determination of viral infectivity in complex animal-related food matrices. Clam and Spanish fermented sausage ("chorizo") samples were spiked with infectious and heat-inactivated human adenovirus-2 (HAdV-2) and mengovirus (vMC). PMA-qPCR/RT-qPCR discriminated infective virus particles, with significant reductions (>2.7 log or 99.7%). Additionally, infectious HAdV-2 and vMC were quantified by plaque assay (in plaque forming units, PFU), and compared with those in virus genomes copies (GCs) quantified by PMA-qPCR/RT-qPCR. A consistent correlation ( > 0.92) was showed between PFU and GCs along serial 10-fold dilutions in both DNA and RNA virus and in both food matrices. This study shows the use of PMA coupled to qPCR/RT-qPCR as a promising alternative for prediction of viral infectivity in food samples in comparison to more expensive and time-consuming methods and for those viruses that are not able to grow under available cell culture techniques.
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http://dx.doi.org/10.3389/fmicb.2016.02008DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5156952PMC
December 2016

Propidium Monoazide Coupled with PCR Predicts Infectivity of Enteric Viruses in Swine Manure and Biofertilized Soil.

Food Environ Virol 2016 Mar 7;8(1):79-85. Epub 2016 Jan 7.

Microbiology Section, Department of Biotechnology and Food Science, Universidad de Burgos, Burgos, Spain.

The use of propidium monoazide (PMA) coupled with real-time PCR (RT-qPCR or qPCR for RNA or DNA viruses, respectively) was assessed to discriminate infectious enteric viruses in swine raw manure, swine effluent from anaerobic biodigester (AB) and biofertilized soils. Those samples were spiked either with infectious and heat-inactivated human adenovirus-2 (HAdV-2) or mengovirus (vMC0), and PMA-qPCR/RT-qPCR allowed discriminating inactivated viruses from the infective particles, with significant reductions (>99.9%). Then, the procedure was further assayed to evaluate the presence and stability of two non-cultivable viruses (porcine adenovirus and rotavirus A) in natural samples (swine raw manure, swine effluent from AB and biofertilized soils); it demonstrated viral inactivation during the storage period at 23 °C. As a result, the combination of PMA coupled to real-time PCR can be a promising alternative for prediction of viral infectivity in comparison to more labour-intensive and costly techniques such as animal or tissue-culture infectivity methods, and for those viruses that do not have currently available cell culture techniques.
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http://dx.doi.org/10.1007/s12560-015-9225-1DOI Listing
March 2016

Human and animal enteric virus in groundwater from deep wells, and recreational and network water.

Environ Sci Pollut Res Int 2015 Dec 25;22(24):20060-6. Epub 2015 Aug 25.

Laboratório de Virologia Aplicada, Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de Santa Catarina, Florianópolis, SC, 88040-900, Brazil.

This study was designed to assess the presence of human adenovirus (HAdV), rotavirus-A (RVA), hepatitis A virus (HAV), and porcine circovirus-2 (PCV2) in groundwater from deep wells, and recreational and network waters. The water samples were collected and concentrated and the virus genomes were assessed and quantified by quantitative PCR (qPCR). Infectious HAdV was evaluated in groundwater and network water samples by integrated cell culture using transcribed messenger RNA (mRNA) (ICC-RT-qPCR). In recreational water samples, HAdV was detected in 100 % (6/6), HAV in 66.6 % (4/6), and RVA in 66.6 % (4/6). In network water, HAdV was detected in 100 % (6/6) of the samples (these 83 % contained infectious HAdV), although HAV and RVA were not detected and PCV2 was not evaluated. In groundwater from deep wells, during rainy period, HAdV and RVA were detected in 80 % (4/5) of the samples, and HAV and PCV2 were not detected; however, during dry period, HAdV and RVA were detected in 60 % (3/5), HAV in only one sample, and PCV2 in 60 % (4/5). In groundwater, all samples contained infectious HAdV. PCV2 presence in groundwater is indicative of contamination caused by swine manure in Concórdia, Santa Catarina, Brazil. The disinfection of human and animal wastes is urgent, since they can contaminate surface and groundwater, being a potential threat for public and animal health.
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http://dx.doi.org/10.1007/s11356-015-5196-xDOI Listing
December 2015

Behaviour and recovery of human adenovirus from tropical sediment under simulated conditions.

Sci Total Environ 2015 Oct 3;530-531:314-322. Epub 2015 Jun 3.

Núcleo de Pesquisas em Agente Emergentes e Re-emegentes, Instituto de Patologia e Saúde Pública, Universidade Federal de Goiás, Brazil. Electronic address:

This study assessed the contributions of pH and organic matter (OM) on the recovery of infectious human adenovirus 5 (HAdV-5) and genome copies (GCs) in waters that were artificially contaminated with tropical soil. The use of a mathematical equation was proposed based on the flocculation index of clay to assess the recovery of total GCs in these controlled assays. The results suggest that solids in the water reduced the viral genome copy loads per millilitre (GC · mL(-1)) and viral infectivity. OM did not influence the GC · mL(-1) recovery rate (p > 0.05) but led to a 99% (2 log10) reduction in plaque-forming unit counts per millilitre (PFU/mL), which indicates that infectivity and gene integrity were non-related parameters. Our findings also suggest that acidic pH levels hinder viral inactivation and that clay is the main factor responsible for the interactions of HAdV-5 with soil. These findings may be useful for future eco-epidemiological investigations and studies of viral inactivation or even as parameters for future research into water quality analysis and water treatment.
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http://dx.doi.org/10.1016/j.scitotenv.2015.05.075DOI Listing
October 2015