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1.
Infect Genet Evol ; 123: 105637, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38986824

ABSTRACT

Viral gastroenteritis is commonly reported in dogs and involves a great diversity of enteric viruses. In this research, viral diversity was investigated in dogs with diarrhea in Northern Brazil using shotgun metagenomics. Furthermore, the presence of norovirus (NoV) was investigated in 282 stool/rectal swabs of young/adult dogs with or without diarrhea from two public kennels, based on one-step reverse transcription polymerase chain reaction (RT-PCR) for genogroup VI and VII (GVI and GVII) and real-time RT-PCR for GI, GII, and GIV. Thirty-one viral families were identified, including bacteriophages. Phylogenetic analyses showed twelve complete or nearly complete genomes belonging to the species of Protoparvovirus carnivoran1, Mamastrovirus 5, Aichivirus A2, Alphacoronavirus 1, and Chipapillomavirus 1. This is the first description of the intestinal virome of dogs in Northern Brazil and the first detection of canine norovirus GVII in the country. These results are important for helping to understand the viral groups that circulate in the canine population.


Subject(s)
Dog Diseases , Gastroenteritis , Phylogeny , Dogs , Animals , Gastroenteritis/virology , Gastroenteritis/veterinary , Gastroenteritis/epidemiology , Brazil/epidemiology , Dog Diseases/virology , Dog Diseases/epidemiology , Norovirus/genetics , Norovirus/classification , Norovirus/isolation & purification , Feces/virology , Genome, Viral , Genetic Variation , Metagenomics/methods , Viruses/classification , Viruses/genetics , Viruses/isolation & purification , Virome
2.
J Appl Microbiol ; 135(5)2024 May 01.
Article in English | MEDLINE | ID: mdl-38627246

ABSTRACT

AIMS: The present study aimed to use a conventional and metagenomic approach to investigate the microbiological diversity of water bodies in a network of drainage channels and rivers located in the central area of the city of Belém, northern Brazil, which is considered one of the largest cities in the Brazilian Amazon. METHODS AND RESULTS: In eight of the analyzed points, both bacterial and viral microbiological indicators of environmental contamination-physical-chemical and metals-were assessed. The bacterial resistance genes, drug resistance mechanisms, and viral viability in the environment were also assessed. A total of 473 families of bacteria and 83 families of viruses were identified. Based on the analysis of metals, the levels of three metals (Cd, Fe, and Mn) were found to be above the recommended acceptable level by local legislation. The levels of the following three physicochemical parameters were also higher than recommended: biochemical oxygen demand, dissolved oxygen, and turbidity. Sixty-three bacterial resistance genes that conferred resistance to 13 different classes of antimicrobials were identified. Further, five mechanisms of antimicrobial resistance were identified and viral viability in the environment was confirmed. CONCLUSIONS: Intense human actions combined with a lack of public policies and poor environmental education of the population cause environmental degradation, especially in water bodies. Thus, urgent interventions are warranted to restore the quality of this precious and scarce asset worldwide.


Subject(s)
Bacteria , Metagenomics , Water Microbiology , Brazil , Bacteria/genetics , Bacteria/isolation & purification , Bacteria/classification , Bacteria/drug effects , Environmental Health , Rivers/microbiology , Rivers/virology , Viruses/genetics , Viruses/isolation & purification , Environmental Monitoring , Drug Resistance, Bacterial/genetics , Humans , Cities , Metals/pharmacology
3.
Article in English | MEDLINE | ID: mdl-38656038

ABSTRACT

Noroviruses are highly infectious, genetically diverse viruses. Global outbreaks occur frequently, making molecular surveillance important for infection monitoring. This cross-sectional descriptive study aimed to monitor cases of norovirus gastroenteritis in the Brazilian Amazon. Fecal samples were tested by immunoenzymatic assay, RT-PCR and genetic sequencing for the ORF1/ORF2 and protease regions. Bayesian inference with a molecular clock was employed to construct the phylogeny. The norovirus prevalence was 25.8%, with a higher positivity rate among children aged 0-24 months. Genogroup GII accounted for 98.1% of the sequenced samples, while GI accounted for 1.9% of them. The GII.P16/GII.4 genotype was the most prevalent, with an evolution rate of 2.87x10-3 and TMRCA estimated in 2012. This study demonstrates that norovirus is a primary causative agent of gastroenteritis and provides data on viral genetic diversity that may facilitate infection surveillance and vaccine development.


Subject(s)
Caliciviridae Infections , Feces , Gastroenteritis , Genotype , Norovirus , Phylogeny , Norovirus/genetics , Norovirus/classification , Brazil/epidemiology , Humans , Caliciviridae Infections/epidemiology , Caliciviridae Infections/virology , Infant , Gastroenteritis/virology , Gastroenteritis/epidemiology , Child, Preschool , Cross-Sectional Studies , Feces/virology , Infant, Newborn , Child , Female , Male , Adolescent , Adult , RNA, Viral/genetics , Prevalence , Young Adult , Reverse Transcriptase Polymerase Chain Reaction , Middle Aged , Aged , Genetic Variation
4.
Article in English | LILACS-Express | LILACS | ID: biblio-1559120

ABSTRACT

ABSTRACT Noroviruses are highly infectious, genetically diverse viruses. Global outbreaks occur frequently, making molecular surveillance important for infection monitoring. This cross-sectional descriptive study aimed to monitor cases of norovirus gastroenteritis in the Brazilian Amazon. Fecal samples were tested by immunoenzymatic assay, RT-PCR and genetic sequencing for the ORF1/ORF2 and protease regions. Bayesian inference with a molecular clock was employed to construct the phylogeny. The norovirus prevalence was 25.8%, with a higher positivity rate among children aged 0-24 months. Genogroup GII accounted for 98.1% of the sequenced samples, while GI accounted for 1.9% of them. The GII.P16/GII.4 genotype was the most prevalent, with an evolution rate of 2.87x10−3 and TMRCA estimated in 2012. This study demonstrates that norovirus is a primary causative agent of gastroenteritis and provides data on viral genetic diversity that may facilitate infection surveillance and vaccine development.

5.
Viruses ; 15(4)2023 04 21.
Article in English | MEDLINE | ID: mdl-37113004

ABSTRACT

Human bocavirus (HBoV) is an emerging virus detected around the world that may be associated with cases of acute gastroenteritis (AGE). However, its contribution to AGE has not been elucidated. This study aimed to describe the frequency, clinical features, and HBoV species circulation in children up to 5 years with or without AGE symptoms in Acre, Northern Brazil. A total of 480 stool samples were collected between January and December 2012. Fecal samples were used for extraction, nested PCR amplification, and sequencing for genotyping. Statistical analysis was applied to verify the association between epidemiological and clinical characteristics. Overall, HBoV-positivity was 10% (48/480), with HBoV-positive rates of 8.4% (19/226) and 11.4% (29/254) recorded in diarrheic and non-diarrheic children, respectively. The most affected children were in the age group ranging between 7 and 24 months (50%). HBoV infection was more frequent in children who live in urban areas (85.4%), use water from public networks (56.2%), and live with adequate sewage facilities (50%). Co-detection with other enteric viruses was 16.7% (8/48) and the most prevalent coinfection was RVA+ HBoV (50%, 4/8). HBoV-1 was the most frequent species detected in diarrheic and non-diarrheic children, responsible for 43.8% (21/48) of cases, followed by HBoV-3 (29.2%, 14/48) and HBoV-2 (25%, 12/48). In this study, HBoV infection was not always associated with AGE, as most HBoV cases belonged to the non-diarrheal group. Future studies are warranted in order to determine the role of HBoV in causing acute diarrhea disease.


Subject(s)
Bocavirus , Gastroenteritis , Human bocavirus , Parvoviridae Infections , Respiratory Tract Infections , Humans , Child , Infant , Child, Preschool , Human bocavirus/genetics , Brazil/epidemiology , Parvoviridae Infections/epidemiology , Gastroenteritis/epidemiology , Diarrhea/epidemiology , Feces , Acute Disease
6.
Braz J Microbiol ; 54(2): 1295-1301, 2023 Jun.
Article in English | MEDLINE | ID: mdl-37076753

ABSTRACT

Viral gastroenteritis is a common clinical problem in dogs and group A rotavirus (RVA) is one of the agents involved in this etiology. It mainly affects dogs in the first 6 months of life, and these animals are considered an important reservoir and potential transmitters of the virus to other susceptible hosts, such as humans. Among the different types of RVA, G3 is the most detected in dogs, and this genotype is also involved in infections in other animals, including humans. Thus, the present study aims to investigate the presence of RVA in samples of dogs from a public kennel. A total of 64 fecal samples from dogs with diarrhea were analyzed, collected from April 2019 to March 2020, from the kennel of the Zoonosis Control Center, located in Belém, a city in the North of Brazil. The extracted genetic material was subjected to reverse transcription followed by real-time PCR (RT-qPCR); the positives were tested by RT-PCR with a specific primer for the RVA VP7 gene, after nucleotide sequencing and phylogenetic analysis. One sample was subjected to high-performance sequencing. A positivity of 7.8% (5/64) was observed for RVA, all characterized as G3, grouping in the G3-III lineage, with greater similarity to human samples. Different regions of the RVA genome fragments were found. These results emphasize the need for animal health surveillance to better understand the global strain dispersion of RVA and elucidate possible interspecies transmission events, monitoring the genetic diversity of this pathogen.


Subject(s)
Gastroenteritis , Rotavirus Infections , Rotavirus , Humans , Dogs , Animals , Rotavirus/genetics , Rotavirus Infections/epidemiology , Rotavirus Infections/veterinary , Gastroenteritis/epidemiology , Gastroenteritis/veterinary , Phylogeny , Brazil/epidemiology , Genome, Viral , Genotype , Feces
7.
Acad Med ; 98(2): 158-161, 2023 02 01.
Article in English | MEDLINE | ID: mdl-35263298

ABSTRACT

The transition from medical student to resident is a pivotal step in the medical education continuum. For applicants, successfully obtaining a residency position is the actualization of a dream after years of training and has life-changing professional and financial implications. These high stakes contribute to a residency application and Match process in the United States that is increasingly complex and dysfunctional, and that does not effectively serve applicants, residency programs, or the public good. In July 2020, the Coalition for Physician Accountability (Coalition) formed the Undergraduate Medical Education-Graduate Medical Education Review Committee (UGRC) to critically assess the overall transition to residency and offer recommendations to solve the growing challenges in the system. In this Invited Commentary, the authors reflect on their experience as the trainee representatives on the UGRC. They emphasize the importance of trainee advocacy in medical education change efforts; reflect on opportunities, concerns, and tensions with the final UGRC recommendations (released in August 2021); discuss factors that may constrain implementation; and call for the medical education community-and the Coalition member organizations in particular-to accelerate fully implementing the UGRC recommendations. By seizing the momentum created by the UGRC, the medical education community can create a reimagined transition to residency that reshapes its approach to training a more diverse, competent, and growth-oriented physician workforce.


Subject(s)
Education, Medical, Undergraduate , Education, Medical , Internship and Residency , Humans , United States , Education, Medical, Graduate , Educational Measurement
8.
PLoS One ; 17(5): e0268566, 2022.
Article in English | MEDLINE | ID: mdl-35594279

ABSTRACT

OBJECTIVES: To characterize the experiences of providers in completing the cause of death section on death certificates, with particular reference to deaths in people who have cancer. METHODS: Focus groups were conducted until thematic saturation was reached, resulting in four groups over three months. Participants were from a variety of specialties and levels and types of training. Focus groups were recorded and transcribed verbatim and analyzed using constant comparison analysis. RESULTS: Three types of challenges to case classification were identified. 1) Infrastructural and procedural challenges encountered when completing death certificates, including the rigid structure of the form, lack of training in its completion, and lack of real-time feedback. 2) Clinical uncertainty and the varied approaches providers take to determine the cause of death based on their perception of the purpose of the death certificate. 3) Choosing cause of death in decedents with a history of cancer. CONCLUSIONS: There are specific and substantial challenges in the death certification process that lead to errors in documenting the cause of death, but many of these challenges could be addressed with structural change to the forms or mechanism of training. Using these data to inform change could improve the death certification process and reliability of this data.


Subject(s)
Clinical Decision-Making , Death Certificates , Cause of Death , Focus Groups , Humans , Reproducibility of Results , Uncertainty
9.
Am J Surg ; 224(1 Pt B): 552-556, 2022 07.
Article in English | MEDLINE | ID: mdl-35164957

ABSTRACT

BACKGROUND: Effective teaching positively impacts student experience during the surgical clerkship. We sought to better understand how medical students characterize excellent surgical educators and how these characteristics may differ between residents and attendings. METHODS: 289 nominations by third-year medical students for a surgical resident and attending teaching award were examined for thematic content using conventional content analysis. RESULTS: Six major themes emerged: inclusion of students, prioritizing student education, facilitating procedural involvement, utilizing effective educational methods, providing mentorship, and role modeling. Residents were more frequently commended for the first three themes, while residents and attendings were recognized equally for the latter three. CONCLUSIONS: In identifying excellent surgical educators, students emphasized the educators' roles in fostering a positive learning environment where student education is prioritized. Residents were recognized more often than attendings for a broader set of qualities valued by students. Residents as teachers training should be structured to develop these qualities.


Subject(s)
Awards and Prizes , Clinical Clerkship , Internship and Residency , Students, Medical , Clinical Clerkship/methods , Humans , Learning , Mentors , Teaching
10.
Future Microbiol ; 16: 847-862, 2021 08.
Article in English | MEDLINE | ID: mdl-34318682

ABSTRACT

Aim: To perform a molecular analysis of rotavirus A (RVA) G3P[6] strains detected in 2012 and 2017 in the Amazon region of Brazil. Materials & methods: Eighteen RVA G3P[6] strains were collected from children aged under 10 years hospitalized with acute gastroenteritis, and partial sequencing of each segment genome was performed using Sanger sequencing. Results: Phylogenetic analysis showed that all G3P[6] strains had a DS-1-like genotype constellation. Two strains had the highest nucleotide identities with equine-like G3P[6]/G3P[8] genotypes. Several amino acid alterations in VP4 and VP7 neutralizing epitopes of equine-like RVA G3P[6] strains were observed in comparison with vaccine strains. Conclusion: These findings suggest that equine-like RVA G3P[6] strains have been circulating in the Amazon region of Brazil as a result of direct importation, and support natural RVA evolutionary mechanisms.


Subject(s)
Genome, Viral , Reassortant Viruses , Rotavirus , Animals , Brazil , Child , Horses , Humans , Phylogeny , Rotavirus/genetics , Rotavirus/isolation & purification
11.
J. Health Biol. Sci. (Online) ; 9(1): 1-6, 2021. tab, graf
Article in English | LILACS | ID: biblio-1352410

ABSTRACT

Objective: We aimed to evaluate the prevalence of intestinal parasitic in the soil of three beaches of Mosqueiro Island, located in the State of Pará, Brazil, as well as to compare the frequency of helminths and protozoa, pathogenic and non-pathogenic parasites according to the beaches analyzed. Methods: This is a cross-sectional analytical study conducted during August and September 2019, in which 155 soil samples were analyzed by Hoffman's method. Results: The results showed that 16.1% of samples were contaminated from 61.3% of collection points. Murubira beach and Farol beach presented the highest prevalence of parasites, however, there was no significant difference between beaches. Also, it was observed a predominance of protozoa (63%) and non-pathogenic parasites (55.6%) in analyzed samples, but there was no statistically significant difference according to the investigated location. Endolimax nana 25.9% (7/27) and hookworms 18.5% (5/27) were the most detected parasites on the beaches. Conclusion: Thus, this study showed parasitic contamination on the beaches from Mosqueiro Island, which may be associated with a lack of sanitation infrastructure and personal hygiene in these places. Therefore, these results reinforce the need to adopt educational and preventive measures to reduce parasitic agents.


Objetivo: Avaliar a prevalência de parasitos intestinais no solo de três praias da Ilha de Mosqueiro, localizada no estado do Pará, Brasil, além de comparar a frequência de helmintos e protozoários, parasitos patogênicos e não patogênicos de acordo com as praias analisadas. Metdodos: Este é um estudo transversal, analítico, realizado durante agosto e setembro de 2019, no qual 155 amostras do solo foram analisadas pelo método de Hoffman. Resultados: Os resultados revelaram que 16,1% das amostras estavam contaminadas em 61,3% dos pontos de coleta. As praias do Murubira e do Farol apresentaram a maior prevalência de parasitos, porém não houve diferença significativa entre os diferentes locais de coleta. Além disso, foi observada a predominância de protozoários (63%) e parasitas não patogênicos (55.6%) nas amostras analisadas, mas não houve diferença estatística significativa entre os locais investigados. Endolimax nana 25,9% (7/27) e Ancilostomídeos 18,5% (5/27) foram os parasitas mais detectados nas praias. Conclusao: Desse modo, este estudo mostrou contaminação parasitária nas praias da Ilha de Mosqueiro, o que pode estar associada à falta de infraestrutura de saneamento e higiene pessoal nesses locais. Além disso, esses resultados reforçam a necessidade de se adotar medidas educacionais e preventivas para a redução desses agentes parasitários.


Subject(s)
Parasitic Diseases , Intestinal Diseases, Parasitic , Parasites , Soil , Sandy Soils , Environmental Pollution
12.
Res Integr Peer Rev ; 5(1): 16, 2020 Dec 01.
Article in English | MEDLINE | ID: mdl-33292815

ABSTRACT

BACKGROUND: Preprint usage is growing rapidly in the life sciences; however, questions remain on the relative quality of preprints when compared to published articles. An objective dimension of quality that is readily measurable is completeness of reporting, as transparency can improve the reader's ability to independently interpret data and reproduce findings. METHODS: In this observational study, we initially compared independent samples of articles published in bioRxiv and in PubMed-indexed journals in 2016 using a quality of reporting questionnaire. After that, we performed paired comparisons between preprints from bioRxiv to their own peer-reviewed versions in journals. RESULTS: Peer-reviewed articles had, on average, higher quality of reporting than preprints, although the difference was small, with absolute differences of 5.0% [95% CI 1.4, 8.6] and 4.7% [95% CI 2.4, 7.0] of reported items in the independent samples and paired sample comparison, respectively. There were larger differences favoring peer-reviewed articles in subjective ratings of how clearly titles and abstracts presented the main findings and how easy it was to locate relevant reporting information. Changes in reporting from preprints to peer-reviewed versions did not correlate with the impact factor of the publication venue or with the time lag from bioRxiv to journal publication. CONCLUSIONS: Our results suggest that, on average, publication in a peer-reviewed journal is associated with improvement in quality of reporting. They also show that quality of reporting in preprints in the life sciences is within a similar range as that of peer-reviewed articles, albeit slightly lower on average, supporting the idea that preprints should be considered valid scientific contributions.

13.
Infect Genet Evol ; 85: 104555, 2020 11.
Article in English | MEDLINE | ID: mdl-32931954

ABSTRACT

Acute gastroenteritis (AG) is responsible for 525,000 deaths worldwide in children under-5-years and is caused by the Human Cosavirus (HCoSV; family Picornaviridae, Genus Cosavirus). Although its health importance, a significant percentage of diarrhea cases (≈ 40 %) still of unknown etiology. In Brazil, few studies have reported HCoSV-A sequences analyzing partial 5' UTR. This study characterized the first near-complete genome of a Cosavirus A (strain AM326) from a child hospitalized with AG in Amazonas state, Northern Brazil. High throughput sequencing (HTS) was performed using the HiSeq™ 2500 platform (Illumina) in one fecal specimen collected from the Surveillance of Rotavirus Network of the Evandro Chagas Institute collected in 2017. Sequence reads were assembled by the De Novo approach using three distinct algorithmic (IDBA-UD, Spades, and MegaHit). The final contig was recovered from the HCoSV-AM326 sample revealing 7,735 nt in length (SRA number SRR12535029; GenBank MT023104) and the genetic characterization, as well as phylogenetic analysis demonstrated a new variant strain from Brazil, highlighting the association of HCoSV-A as a possible causative agent of AG. This finding demonstrates the importance of the metagenomic approach to elucidate cases of diarrhea without a defined etiology, as well as providing a better understanding about the virus genetics, evolution and epidemiology.


Subject(s)
Gastroenteritis/diagnosis , Gastroenteritis/virology , Picornaviridae Infections/diagnosis , Picornaviridae Infections/virology , Picornaviridae/classification , Picornaviridae/genetics , Acute Disease , Brazil , Child , Genome, Viral , Genomics/methods , High-Throughput Nucleotide Sequencing , Hospitalization , Humans , Picornaviridae/isolation & purification , RNA, Viral
14.
J Med Virol ; 91(12): 2125-2133, 2019 12.
Article in English | MEDLINE | ID: mdl-31429939

ABSTRACT

Immunosuppressive therapy causes severe impairment of host defense and diarrhea is a frequent complication in renal transplant recipients. This study aimed to describe the occurrence of Rotavirus A (RVA) and Human Bocavirus (HBoV) in fecal samples of immunosuppressed patients submitted to renal transplantation during posttransplant follow-up. A longitudinal study was carried out involving a 25-patient cohort, selected for kidney transplantation. A total of 126 fecal samples were collected between May 2014 and May 2016. Molecular techniques were used to detect and characterize circulating RVA and HBoV genotypes and statistical analysis were applied to verify the association between epidemiological and clinical characteristics. The prevalence of RVA and HBoV was 24% (6/25) and 40% (10/25), respectively. Among RVA and HBoV positive cases, the majority was female; did not conduct water treatment nor had adequate sewage facilities. The most detected genotypes were RVA G3 (62.5%) and HBoV-3 (95%). Phylogenetic analysis of HBoV strains indicated that studied samples were similar to those found in Asian and American countries. The present study point out the circulation of these viral agents among immunosuppressed individuals and these findings will enable the construction of new knowledge and care perspectives on the cause of diarrhea in this population.


Subject(s)
Feces/virology , Human bocavirus/isolation & purification , Immunocompromised Host , Kidney Transplantation/adverse effects , Rotavirus/isolation & purification , Adult , Brazil/epidemiology , Coinfection/epidemiology , Coinfection/virology , Female , Genotype , Human bocavirus/genetics , Humans , Longitudinal Studies , Male , Parvoviridae Infections/epidemiology , Parvoviridae Infections/virology , Phylogeny , Prevalence , Rotavirus/genetics , Rotavirus Infections/epidemiology , Rotavirus Infections/virology , Sequence Analysis, DNA , Transplant Recipients/statistics & numerical data
15.
J Med Microbiol ; 68(8): 1233-1239, 2019 Aug.
Article in English | MEDLINE | ID: mdl-31215861

ABSTRACT

PURPOSE: Human bocavirus (HBoV) is a DNA virus that is mostly associated with respiratory infections. However, because it has been found in stool samples, it has been suggested that it may be a causative agent for human enteric conditions. This underpins the continuous search for HBoVs, especially after the introduction of the rotavirus vaccine due to acute gastroenteritis cases related to emergent viruses, as HBoVs are more likely to be found in this post-vaccine scenario. Therefore, the aim of this study is to demonstrate the prevalence of HBoV in children aged less than 10 years with acute gastroenteritis in Brazil from November 2011 to November 2012. METHODOLOGY: Stool samples from hospitalized children ≤10 years old who presented symptoms of acute gastroenteritis were analysed for the presence of rotavirus A (RVA) by an enzyme-linked immunosorbent assay (ELISA), and for HBoV DNA by nested PCR. RESULTS: HBoV positivity was detected in 24.0 % (54/225) of samples. Two peaks of HBoV detection were observed in November 2011 and from July to September 2012. Co-infections between HBoV and rotavirus A were identified in 50.0 % (27/54) of specimens. Phylogenetic analysis identified the presence of HBoV-1 (94.8 %), HBoV-2 (2.6 %) and HBoV-3 (2.6 %) species, with only minor variations among them. CONCLUSION: Our findings provide evidence for the circulation of most HBoV genotypes (except HBoV-4) in the North Region of Brazil at a considerable rate and further investigations are necessary to improve our knowledge in the context of HBoV infections and their role in gastrointestinal diseases.


Subject(s)
Gastroenteritis/epidemiology , Human bocavirus/genetics , Molecular Epidemiology , Parvoviridae Infections/epidemiology , Acute Disease , Brazil/epidemiology , Capsid Proteins/genetics , Child , Child, Preschool , Coinfection/epidemiology , Coinfection/virology , DNA, Viral/genetics , Feces/virology , Female , Gastroenteritis/virology , Genotype , Human bocavirus/classification , Human bocavirus/isolation & purification , Humans , Infant , Infant, Newborn , Male , Parvoviridae Infections/virology , Phylogeny , Polymerase Chain Reaction , Prevalence , Rotavirus/immunology , Rotavirus/isolation & purification , Rotavirus Infections/epidemiology , Rotavirus Infections/virology , Sequence Analysis, DNA
16.
BMC Pediatr ; 19(1): 193, 2019 06 12.
Article in English | MEDLINE | ID: mdl-31189470

ABSTRACT

BACKGROUND: Rotavirus antigenemia and RNAemia (the presence of rotavirus RNA in serum) have been commonly identified among paediatric patients with acute gastroenteritis. In this study we examined the association between rotavirus antigenemia and clinical features, and sought to determine the genotypes of rotaviruses detected in paired stool and serum samples. METHODS: Paired stool and serum samples were obtained from children hospitalised for acute gastroenteritis in Belém, Brazil, between June 2012 and June 2015. The 20-point Vesikari scoring system was used to assess the disease severity upon a retrospective medical record review. Stool and serum samples were primarily screened for the presence of rotavirus antigen using a commercial ELISA assay. The rotavirus isolates from stool and serum samples were genotyped by using the classical reverse-transcriptase polymerase chain reaction (RT-PCR) and/or through nucleotide sequencing of VP4 and VP7 genes. Viral load was estimated using real-time RT-PCR. RESULTS: In total rotavirus antigen was detected in 109 (24.2%) stool samples from 451 children, whereas antigenemia occurred in 38.5% (42/109) of these patients. We demonstrated that patients positive for rotavirus RNA in paired stool and serum samples were more likely to have a higher frequency of vomiting episodes in a 24-h period (p = 0.0035). Our findings also suggested that children not vaccinated against rotavirus are more likely to develop antigenemia, as compared to those given at least one vaccine dose (p = 0.0151). G12P [8] and G2P [4] genotypes were predominant throughout the study period, accounting for 52.3% (57/109) and 27.5% (30/109) of the typed isolates, respectively. Ten stool-serum pairs could be typed for VP4 and VP7 genes. Seven of these pairs showed concordant results with G2P [4] genotype being detected in stool and serum samples, whereas discrepancies between genotypes (G2P [4]/G2P[NT] and G12P [8]/G2P[NT]) were seen in three pairs. CONCLUSIONS: Rotavirus antigenemia and RNAemia occur in a significant number of children hospitalised for acute gastroenteritis in Belém, Brazil, and may contribute to a greater disease severity, particularly translated into a greater number of vomiting episodes. This study documented a high concordance of genotypes detected in a subgroup of paired stool and serum samples.


Subject(s)
Antigens, Viral/analysis , Gastroenteritis/immunology , RNA, Viral/analysis , Rotavirus Infections/immunology , Rotavirus/immunology , Acute Disease , Antigens, Viral/blood , Brazil , Child , Child, Preschool , Enzyme-Linked Immunosorbent Assay , Feces/chemistry , Feces/virology , Female , Gastroenteritis/complications , Gastroenteritis/virology , Genotype , Hospitalization , Humans , Male , Prospective Studies , RNA, Viral/blood , Reverse Transcriptase Polymerase Chain Reaction , Rotavirus/genetics , Severity of Illness Index , Vomiting/etiology
17.
Arch Virol ; 164(8): 2107-2117, 2019 Aug.
Article in English | MEDLINE | ID: mdl-31144039

ABSTRACT

Species A rotavirus still remains a major cause of acute gastroenteritis in infants and young children. Globally, six genotypes (G1P[8], G2P[4], G3P[8], G4P[8], G9P[8] and G12P[8]) account for >90% of circulating strains; however, genotype G12 in combination with P[6] or P[9] has been detected at increasing rates. We sought to broaden our knowledge about the rotavirus strains circulating during the early post-vaccine-introduction period. Stool samples were obtained from children hospitalised for acute gastroenteritis in Belém, Northern Brazil, from May 2008 to May 2011 and examined by reverse transcription polymerase chain reaction and nucleotide sequencing. A total of 122 out of the original 1076 rotavirus strains were judged to be non-typeable in the first analysis and were therefore re-examined. G2P[4] was the most prevalent genotype (58.0%), followed by G1P[8] (16.9%), and G12P[6] (7.5%). G12P[6] strains were identified at similar rates during the first (2.5%) and second (3.9%) years, and the rate jumped to 15.6% in the third year. Analysis of VP7 sequences of the G12P[6] strains showed that they belonged to lineage III. In addition, co-circulating G12P[6] strains displaying long and short RNA patterns were found to belong to the Wa-like and DS-1-like constellation, respectively. Additional unusual circulating strains G12P[9] and G3P[9] were also identified. This hospital-based study showed a high prevalence of G12P[6] strains in the third year of surveillance. Our results highlight the need for continuous longitudinal monitoring of circulating rotavirus strains after introduction of rotavirus vaccines in Brazil and elsewhere.


Subject(s)
Gastroenteritis/virology , Rotavirus/genetics , Antigens, Viral/immunology , Brazil , Child , Child, Hospitalized , Gastroenteritis/immunology , Genotype , Humans , Molecular Epidemiology/methods , Phylogeny , Prevalence , RNA, Viral/genetics , Rotavirus/immunology , Rotavirus Infections/immunology , Rotavirus Infections/virology , Rotavirus Vaccines/immunology , Sequence Analysis, DNA/methods
19.
PLoS One ; 13(12): e0209005, 2018.
Article in English | MEDLINE | ID: mdl-30562373

ABSTRACT

Acute gastroenteritis is one of the main causes of mortality in humans and young animals. Domestic and mainly wild animals such as bats, small rodents and birds are highly diversified animals in relation to their habitats and ecological niches and are widely distributed geographically in environments of forest fragmentation in some areas of the Amazon, being considered important sources for viruses that affect humans and other animals. Due to the anthropical activities, these animals changed their natural habitat and adapted to urbanized environments, thus representing risks to human and animal health. Although the knowledge of the global diversity of enteric viruses is scarce, there are reports demonstrating the detection of rotavirus in domestic animals and animals of productive systems, such as bovines and pigs. The present study investigated the prevalence of Rotavirus A in 648 fecal samples of different animal species from the northeastern mesoregion of the state of Pará, Brazil, which is characterized as an urbanized area with forest fragments. The fecal specimens were collected from October 2014 to April 2016 and subjected to a Qualitative Real-Time Polymerase Chain Reaction (RT-qPCR), using the NSP3 gene as a target. It was observed that 27.5% (178/648) of the samples presented positive results for RVA, with 178 samples distributed in birds (23.6%), canines (21.35%), chiropterans (17.98%), bovines (14.6%), horses (8.43%), small rodents (6.74%), pigs (3.93%) and felines (3.37%), demonstrating the circulation of RVA in domestic animals and suggesting that such proximity could cause transmissions between different species and the occurrence of rearrangements in the genome of RVA as already described in the literature, associated to the traces of environmental degradation in the studied areas.


Subject(s)
Animals, Domestic/virology , Animals, Wild/virology , Forests , Rotavirus Infections/veterinary , Rotavirus , Animals , Brazil , Cities , Feces/virology , Rotavirus Infections/epidemiology , Urbanization
20.
J Med Virol ; 89(6): 974-981, 2017 06.
Article in English | MEDLINE | ID: mdl-27862014

ABSTRACT

The species A rotaviruses (RVA) are important gastroenteric pathogens that infect humans and animals. RVA genotype G3P[9] has been described in human-animal reassortment events, and the complexity of its hosts motivates the genetic investigation of this strain. Therefore, the aim of this study is to analyse a G3P[9] sample that was detected in a child with acute gastroenteritis. The 1A3739 sample featured the constellation G3P[9]-I18-R3-C3-Mx-A19-N3-T3-E3-H6. The sequence for VP3 gene was not obtained. The phylogeny showed a closer relationship among genes VP7, VP1, NSP3, NSP4, and NSP5 with genes of animal origin, such as chiropter, alpaca, equine, and simian. In addition, the genes VP6 and NSP1 belong to the new genotypes I18 and A19, respectively. The emergence of strains such as these can interfere with the effectiveness of the RVA vaccine, and continuous monitoring is therefore important. Additional studies are needed to determine the evolutionary source and to identify a possible reservoir of RVA in nature.


Subject(s)
Gastroenteritis/virology , Genotype , Reassortant Viruses/genetics , Rotavirus Infections/virology , Rotavirus/genetics , Child, Preschool , Evolution, Molecular , Female , Humans , Phylogeny , Recombination, Genetic , Rotavirus/isolation & purification , Sequence Analysis, DNA
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