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SUMMARY OBJECTIVE: Genome sequencing has been proved to be an excellent tool to monitor the molecular epidemiology of the disease caused by severe acute respiratory syndrome coronavirus 2, i.e., coronavirus disease 2019. Some reports of infected, vaccinated individuals have aroused great interest because they are primarily being infected with circulating variants of concern. To investigate the cases of infected, vaccinated individuals in Salvador, Bahia, Brazil, we performed genomic monitoring to estimate the magnitude of the different variants of concern in these cases. METHODS: Nasopharyngeal swabs from infected (symptomatic and asymptomatic), vaccinated or unvaccinated individuals (n=29), and quantitative reverse transcription polymerase chain reaction cycle threshold value (Ct values) of ≤30 were subjected to viral sequencing using nanopore technology. RESULTS: Our analysis revealed that the Omicron variant was found in 99% of cases and the Delta variant was found in only one case. Infected, fully vaccinated patients have a favorable clinical prognosis; however, within the community, they become viral carriers with the aggravating factor of viral dissemination of variants of concern not neutralized by the currently available vaccines. CONCLUSION: It is important to acknowledge the limitations of these vaccines and to develop new vaccines to emergent variants of concern, as is the case of influenza vaccine; going through new doses of the same coronavirus vaccines is “more of the same.”

BACKGROUND: There is growing concern about individuals reported to suffer repeat COVID-19 disease episodes, these in a small number of cases characterised as de novo infections with distinct sequences, indicative of insufficient protective immunity even in the short term. METHODS: Observational case series and case-control studies reporting 33 cases of recurrent, symptomatic, qRT-PCR positive COVID-19. Recurrent disease was defined as symptomatic recurrence after symptom-free clinical recovery, with release from isolation >14 days from the beginning of symptoms confirmed by qRT-PCR. The case control study-design compared this group of patients with a control group of 62 patients randomly selected from the same COVID-19 database. RESULTS: Of 33 recurrent COVID-19 patients, 26 were female and 30 were HCW. Mean time to recurrence was 50.5 days which was associated with being a HCW (OR 36.4 (p

Zika virus (ZIKV) is considered to cause an acute self-limited infection in adults, and microcephaly in fetus. Presence of the virus for long periods has been detected in body fluids; however, persistent viremia in serum for more than 1 year has not yet been reported. We have investigated persistence of ZIKV in serum samples of 77 subjects who were infected by the virus between 18 months and 3 years before the start of this study. The subjects included children with microcephaly and their parents. Serum samples were subjected to routine RT-qPCR assay for ZIKV, Chikungunya virus, and Dengue virus. From the 77 subjects, five showed positive for the presence of ZIKV particles by RT-qPCR, including four members of the same family. Viral isolation in Vero cells and C6/36 cells confirmed the result and showed the viral particles were active. We have detected viremia in healthy carriers up to 3 years after symptom onset. Humans acting as potential viral reservoirs have major implication for the current understanding of ZIKV infection.

Genome sequencing proved to be an excellent tool to monitor the molecular epidemiology of the disease caused by SARS-CoV-2, i.e., coronavirus disease (COVID-19). Some reports of infected, vaccinated individuals have aroused great interest because they are primarily being infected with circulating variants of concern (VOCs). To investigate the cases of infected, vaccinated individuals in Salvador, Bahia, Brazil, we performed genomic monitoring to estimate the magnitude of the different VOCs in these cases. Nasopharyngeal swabs from infected (symptomatic and asymptomatic), fully vaccinated individuals (n=29) who were of varying age and had RT-qPCR Ct values of ≤30 were subjected to viral sequencing using Nanopore technology. Our analysis revealed that the Omicron variant was found in 99% of cases and that only one case was due to the Delta variant. Infected, fully vaccinated patients have a favorable clinical prognosis; however, within the community, they become viral carriers with the aggravating factor of viral dissemination of VOCs not neutralized by the vaccines.

The etiology of viral meningoencephalitis is frequently unidentified. Chikungunya virus (CHIKV) and Zika virus (ZIKV) are known to affect the central nervous system and should therefore be considered in the diagnosis of meningoencephalitis, as its outcome may be influenced by the etiologic agent, age, and immunological condition of the patient. In this study, we aimed to determine whether CHIKV and ZIKV were the etiological agents of viral encephalitis in patients with meningoencephalitis admitted to the main hospital of infectious diseases in the city of Salvador, Brazil. Of the 1,049 patients with neurological symptoms who were admitted to the hospital during the study period, 149 were enrolled and 20 (13.34%) tested positive for ZIKV (12%) or CHIKV (1.34%). No specific clinical manifestations were observed to be associated with ZIKV or CHIKV infections. Determination of the etiological agent of meningitis and encephalitis is important for patient management and appropriate treatment.

The outbreak of the new coronavirus (SARS-CoV-2) causing the coronavirus disease (COVID-19) has spread globally. As of June 18, 2020, a high maternal mortality rate due to SARS-CoV-2 infections was identified in Brazil, representing most of the world cases at that time. An observational, cross-sectional study was performed with pregnant women admitted in two maternity hospitals located in Salvador/Bahia and their newborns, from May 24th up to July 17th of 2020. Among 329 pregnant women enrolled at hospital admission, a high prevalence (n=28; 8.5%) of pregnant women with COVID-19 was observed, as well as a high proportion of asymptomatic cases (n=19; 67.9%). Two newborns had detectable SARS-CoV-2 but evolved without abnormalities. This data highlight the importance of identifying pregnant women with COVID-19 for proper isolation measures to prevent in-hospital transmission.

Arboviruses, including the Zika virus, have recently emerged as one of the most important threats to human health. The use of metagenomics-based approaches has already proven valuable to aid surveillance of arboviral infections, and the ability to reconstruct complete viral genomes from metatranscriptomics data is key to the development of new control strategies for these diseases. Herein, we used RNA-based metatranscriptomics associated with Ion Torrent deep sequencing to allow for the high-quality reconstitution of an outbreak-related Zika virus (ZIKV) genome (10,739 nt), with extended 5′-UTR and 3′-UTR regions, using a newly-implemented bioinformatics approach. Besides allowing for the assembly of one of the largest complete ZIKV genomes to date, our strategy also yielded high-quality complete genomes of two arthropod-infecting viruses co-infecting C6/36 cell lines, namely: Alphamesonivirus 1 strain Salvador (20,194 nt) and Aedes albopictus totivirus-like (4618 nt); the latter likely represents a new viral species. Altogether, our results demonstrate that our bioinformatics approach associated with Ion Torrent sequencing allows for the high-quality reconstruction of known and unknown viral genomes, overcoming the main limitation of RNA deep sequencing for virus identification.

Highlights • Detection of a SARS-CoV-2 genome through metatranscriptome next-generation sequencing. • Direct metatranscritptome sequencing from nasopharyngeal swab of a COVID-19 patient. • Depletion of human rRNA and in-house bioinformatics strategy for Ion torrent sequencing., Herein, we describe the detection of a SARS-CoV-2 genome through metatranscriptome next-generation sequencing directly from the nasopharyngeal swab of a suspected case of local transmission of Covid-19, in Brazil. Depletion of human ribosomal RNA and use of an optimized in-house developed bioinformatics strategy contributed to successful detection of the virus.

Background Chikungunya is an arbovirus, transmitted by Aedes mosquitoes, which emerged in the Americas in 2013 and spread rapidly to almost every country on this continent. In Brazil, where the first cases were detected in 2014, it currently has reached all regions of this country and more than 900,000 cases were reported. The clinical spectrum of chikungunya ranges from an acute self-limiting form to disabling chronic forms. The purpose of this study was to estimate the seroprevalence of chikungunya infection in a large Brazilian city and investigate the association between viral circulation and living condition. Methodology/principal findings We conducted a population-based ecological study in selected Sentinel Areas (SA) through household interviews and a serologic survey in 2016/2017. The sample was of 1,981 individuals randomly selected. The CHIKV seroprevalence was 22.1% (17.1 IgG, 2.3 IgM, and 1.4 IgG and IgM) and varied between SA from 2.0% to 70.5%. The seroprevalence was significantly lower in SA with high living conditions compared to SA with low living condition. There was a positive association between CHIKV seroprevalence and population density (r = 0.2389; p = 0.02033). Conclusions/significance The seroprevalence in this city was 2.6 times lower than the 57% observed in a study conducted in the epicentre of the CHIKV epidemic of this same urban centre. So, the herd immunity in this general population, after four years of circulation of this agent is relatively low. It indicates that CHIKV transmission may persist in that city, either in endemic form or in the form of a new epidemic, because the vector infestation is persistent. Besides, the significantly lower seroprevalences in SA of higher Living Condition suggest that beyond the surveillance of the disease, vector control and specific actions of basic sanitation, the reduction of the incidence of this infection also depends on the improvement of the general living conditions of the population., Author summary Chikungunya is a disease caused by viruses transmitted by Aedes mosquitoes. It stands out because it can produce chronic cases and disabling forms. This survey was conducted in a large city in Brazil, involving 1,981 people living in different living conditions/LC, distributed in 30 urban areas. Interviews and blood collection were performed to know the seroprevalence of chikungunya virus (CHIKV). The global seroprevalence was 22.1%, ranged from 2.0 to 70.5%. The areas where the highest and lowest seroprevalence were of low LC, but population density was different among them, which was related to seroprevalence. The mosquito index was not associated with the seroprevalence. Our results showed that lower seroprevalence than another survey conducted in the epicentre of the epidemic of this same city. So, a considerable proportion of the population of Feira de Santana is susceptible to the CHIKV. Possibly the virus will continue to circulate and manifest itself as endemic or epidemic, as long as LC remain favourable to the reproduction of the transmitting vector of CHIKV.

Congenital Zika virus syndrome (CZS) is associated with damage to neural progenitor cells by ZIKA virus infection. There are no accurate statistics on the percentage of pregnant mothers who have had babies affected by the syndrome. Few cases of discordant twins have been described in the literature and, therefore, we hypothesize that the genetic background of the progeny and/or mother may play a role in the fate of the syndrome. We performed a complete exome sequencing in a set of dizygotic individuals and their parents. After that, we selected discordant variants on the MTOR gene between the affected and unaffected twin and we observed a mutation (rs2295079), placed in a region restricted to proximal 5′-UTR, as a strong possible causal variant. In addition, in most brain tissues (including fetal brain) evaluated for expression quantitative trait loci (eQTL), this locus is strongly correlated with post-translational modifications of histones (promoter and enhancer marks) and hypersensitivity to DNAse I (open chromatin mark). Taken together, our data suggest that changes in the MTOR gene may be related to CZS. Additional functional studies should be carried out to prove how and why a MTOR mutation can predispose the fetus to the syndrome., Zika Virus; mTOR; Whole-exome sequencing; Twins; polymorphisms

University of California San Francisco. San Francisco, California, USA. University of Oxford. Oxford, UK. Federal University of Bahia. Salvador, BA, Brasil. University of California San Francisco. San Francisco, California, USA. University of Washington. Seattle, Washington, USA. Hospital Alian?a. Salvador, BA, Brasil. Federal University of Bahia. Salvador, BA, Brasil. Funda??o Gon?alo Moniz. Centro de Pesquisas Gon?alo Moniz. Salvador, BA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. University of Oxford. Oxford, UK. University of California San Francisco. San Francisco, California, USA. Sequencing of isolates from patients in Bahia, Brazil, where most Zika virus cases in Brazil have been reported, resulted in 11 whole and partial Zika virus genomes. Phylogenetic analyses revealed a well-supported Bahia-specific Zika virus lineage, which indicates sustained Zika virus circulation in Salvador, Bahia's capital city, since mid-2014 .

Metagenomic next-generation sequencing (mNGS) of samples from 15 patients with documented Zika virus (ZIKV) infection in Bahia, Brazil, from April 2015 to January 2016 identified coinfections with chikungunya virus (CHIKV) in 2 of 15 ZIKV-positive cases by PCR (13.3%). While generally nonspecific, the clinical presentation corresponding to these two CHIKV/ZIKV coinfections reflected infection by the virus present at a higher titer. Aside from CHIKV and ZIKV, coinfections of other viral pathogens were not detected. The mNGS approach is promising for differential diagnosis of acute febrile illness and identification of coinfections, although targeted arbovirus screening may be sufficient in the current ZIKV outbreak setting.

Chikungunya virus (CHIKV) has never been detected in human breast milk. This is a brief report of CHIKV infection in a breastfeeding woman of a 3-month-old baby. The mother's CHIKV-RT PCR was positive in serum, urine and milk. The baby's CHIKV serology and reverse transcription polimerase chain reaction (RT-PCR) were negative. The detection of CHIKV in milk raises clinical and epidemiologic questions.

Epidemiological surveillance for Human Bocavirus (HBoV) was conducted on 105 fecal specimens from children with acute gastroenteritis in Bahia, Brazil. Among of a total 105 stool samples, 44 samples were positive for HBoV as detected by nested-PCR. Of the 44 positive samples, co-infections with other enteric viruses (Norovirus, Adenovirus, and Rotavirus) were found in 12 pediatric patients. Mixed infections among HBoV with Norovirus were frequently observed in this population. The phylogenetic analysis identified the presence of HBoV-1, and HBoV 2A species. This study shows that HBoV is another viral pathogen in the etiology of acute gastroenteritis in children in Bahia, Brazil. J. Med. Virol. © 2015 Wiley Periodicals, Inc.

Zika and dengue viruses are closely related flaviviruses, with immunological interactions and identical urban, mosquito-borne transmission.1Therefore, the recent introduction of Zika virus into the Americas and large-scale exposure of a uniformly previously unexposed population could affect subsequent transmission of dengue virus. This hypothesis had been untested, largely because sufficient epidemiological data were not available from affected locations. We explored this hypothesis in Salvador, the fourth largest city in Brazil (population 2·9 million), where extensive transmission of dengue viruses 1–42,3occurred before the introduction and spread of Zika virus in 2015. Fil: Ribeiro, Guilherme Sousa. Universidade Federal da Bahia; Brasil. Fundación Oswaldo Cruz; Brasil Fil: Kikuti, Mariana. Fundación Oswaldo Cruz; Brasil. Universidade Federal da Bahia; Brasil Fil: Tauro, Laura Beatriz. Fundación Oswaldo Cruz; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú | Universidad Nacional de Misiones. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú; Argentina Fil: Nascimento, Leile Camila J.. Fundación Oswaldo Cruz; Brasil Fil: Cardoso, Cristiane W.. Secretaria Municipal de Saúde de Salvador; Brasil Fil: Campos, Gúbio S.. Universidade Federal da Bahia; Brasil Fil: Ko, Albert I.. Fundación Oswaldo Cruz; Brasil. University of Yale; Estados Unidos Fil: Weaver, Scott C.. University of Texas Medical Branch; Estados Unidos Fil: Reis, Mitermayer G.. Fundación Oswaldo Cruz; Brasil. Universidade Federal da Bahia; Brasil Fil: Kitron, Uriel D.. University of Emory; Estados Unidos. Fundación Oswaldo Cruz; Brasil Fil: Paploski, Igor A. D.. Universidade Federal da Bahia; Brasil Fil: Silva, Monaise M. O.. Fundación Oswaldo Cruz; Brasil Fil: Kasper, Amelia M.. Universidade Federal da Bahia; Brasil Fil: Tavares, Aline S.. Fundación Oswaldo Cruz; Brasil Fil: Cruz, Jaqueline S.. Fundación Oswaldo Cruz; Brasil Fil: Moreira, Patrícia S. S.. Universidade Federal da Bahia; Brasil Fil: Anjos, Rosângela O.. Fundación Oswaldo Cruz; Brasil Fil: Araújo, Josélio M. G.. Universidade Federal do Rio Grande do Norte; Brasil Fil: Khouri, Ricardo. Fundación Oswaldo Cruz; Brasil Fil: Sardi, Silvia I.. Universidade Federal da Bahia; Brasil

Chikungunya virus (CHIKV) is an RNA virus transmitted by Aedes mosquitoes. The clinical manifestations include fever, arthralgia, rash, and other atypical clinical findings including ocular lesions. We report the case of a 57-year-old man with meningoencephalitis and anterior uveitis due to CHIKV. The patient had developed bilateral anterior uveitis with iris atrophy and a cotton wool spot on the left eye, and his serum, urine, saliva, and cerebrospinal fluid were positive for CHIKV by RT-PCR. The spectrum of the ophthalmologic manifestations and its pathophysiology in cases of CHIKV infections needs to be better understood. Additional studies examining the ocular lesions caused by CHIKV could improve the therapeutic goals of reducing the morbidity and sequels.

We describe a series of 15 Haff disease cases from an outbreak in Salvador, Brazil, starting early December 2016. Eleven cases were grouped in four family clusters of two to four individuals, four were isolated cases. All but one patient consumed cooked fish; 11 within 24h before symptoms onset. Cases consumed ‘Olho de Boi’ (Seriola spp.) and ’Badejo’ (Mycteroperca spp.). A total of 67 cases were detected, the last case was reported on 5 April 2017.

Primary and continuous cell culture systems are a good in vitro alternative for the quantitative assessment of snake venom toxicity. There are still few studies evaluating the cellular response against Bothrops leucurus venom (BlV). This study aimed to evaluate the cytotoxic effect of crude BlV on culture of VERO E6 cells. Neutral Red incorporation (NR) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) tests were performed for viable cells, and Trypan Blue (TB) dye exclusion test was performed for non-viable cells. The differences between the groups were evaluated by ANOVA-One-Way and Dunnet through the program Graph Pad 5.0 with significance of 5%. The 50% cytotoxic concentration (CC50) of BlV was 7.86 μg/mL. There was a trend of dose-dependent reduction on cell viability in all assays evaluated. However, MTT and NR tests were more sensitive for the evaluation of BlV cytotoxicity on the VERO E6 cell line.

Absract We report the presence of Chikungunya (CHIKV) RNA in the blood, urine and semen during the acute phase of the disease in an adult with a dual infection with Dengue virus type 3. The patient, a 25 yr-old man from Salvador, Brazil, reported a 6-day duration of high fever, arthralgia, myalgia, headache and photophobia.Blood and semen specimens were positive for CHIKV in the first collected samples; semen and urine specimens were positive for CHIKV after 30 days of symptoms onset. DENV-3 RNA was positive in blood specimen when first collected 6 days after the initiation of symptoms.We describe for the first time the presence of CHIKV RNA in urine and semen for an extended period of time and we address the possible implications of these findings for diagnosis and transmission dynamics.

Equine infectious anemia is an important infectious disease that affects equids worldwide. Control of the disease is currently based on detection of anti-p26 EIAV by Agar Gel Immunodiffusion (AGID). In this work, 62 animals were examined by AGID and nested-PCR using primers for the gag gene. Fifty-three samples (85.5%) were positive by nested-PCR, whereas only 33 samples (53%) were positive for AGID. Fifteen amplicons obtained by nested-PCR were sequenced and the aligned results subjected to phylogenetic analysis. The analysis suggests that the Brazilian EIAV form a cluster with WSU5, EIAVUK and Wyoming strains from United States.

Metagenomic next-generation sequencing coupled with capture probe enrichment was used to recover 11 whole and partial Zika virus (ZIKV) genomes from patients in Bahia, Brazil from April 2015 to January 2016, where the majority of suspected Brazilian ZIKV cases have been reported. Phylogenetic reconstructions and molecular clock analyses using the newly generated data uncovered the existence of a Bahia-specific ZIKV lineage sharing a common ancestor in mid-2014, indicating sustained circulation of this strain in Bahia since that date.

CAPES (Zika FastTrack), FINEP, FAPERJ and FAPESB. Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil Federal University of Bahia. Laboratory of Virology, Salvador, BA, Brasil Federal University of Bahia. Laboratory of Virology, Salvador, BA, Brasil Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia. Department of Pathology. Salvador, BA, Brasil Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil Hospital Couto Maia. Salvador, BA, Brasil Federal University of Rio de Janeiro. Institute of Biophysics Carlos Chagas Filho. Rio de Janeiro, RJ, Brasil São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil Zika virus (ZIKV) infection has been associated with severe complications both in the developing and adult nervous system. To investigate the deleterious effects of ZIKV infection, we used human neural progenitor cells (NPC), derived from induced pluripotent stem cells (iPSC). We found that NPC are highly susceptible to ZIKV and the infection results in cell death. ZIKV infection led to a marked reduction in cell proliferation, ultrastructural alterations and induction of autophagy. Induction of apoptosis of Sox2(+) cells was demonstrated by activation of caspases 3/7, 8 and 9, and by ultrastructural and flow cytometry analyses. ZIKV-induced death of Sox2(+) cells was prevented by incubation with the pan-caspase inhibitor, Z-VAD-FMK. By confocal microscopy analysis we found an increased number of cells with supernumerary centrosomes. Live imaging showed a significant increase in mitosis abnormalities, including multipolar spindle, chromosome laggards, micronuclei and death of progeny after cell division. FISH analysis for chromosomes 12 and 17 showed increased frequency of aneuploidy, such as monosomy, trisomy and polyploidy. Our study reinforces the link between ZIKV and abnormalities in the developing human brain, including microcephaly.

This work was carried out to characterize and evaluate five bovine rotavirus, monoclonal antibodies (MAbs), as a diagnosis tool, by isotyping, dot-blot, western-blot, indirect immunofluorescence (IFI) and ELISA techniques. The immunochemistry characterization showed that all five MAbs (4F7, 4F3, 1G5, 1E12 and 3C12) were IgG2a isotype. The dot-blot immunoassay showed that 1G5, 4F7, 1E12 and 4F3 detected viral antigen in different concentrations and two MAbs (1E12 and 4F3) recognized viral proteins by western-blot. All MAbs detected viral antigen in bovine and human fecal samples by capture ELISA technique and viral antigen in infected MA-104 cell culture by IFI. In conclusion, two groups of Mabs were indetified: one with Mabs 4F7, 1E12 and 1G5 showed the best results to detect rotavirus antigen in fecal samples by capture ELISA or dot-blot techniques assay and other with 4F3 and 3C12 which may be used to detect rotavirus antigens in cell culture by IFI. The results showed the potential use of these MAbs as diagnosis tools in diarrheas by rotavirus in bovines.

To the Editor: Zika virus (ZIKV) has been recognized as an emerging mosquito-borne flavivirus since outbreaks were reported from Yap Island in 2007 (1), French Polynesia in 2013 (2), and Cook Island and New Caledonia in 2014 (3). It has joined dengue virus (DENV) and chikungunya virus (CHIKV) as global public health threats (4). ZIKV infection typically causes a self-limited dengue-like illness characterized by exanthema, low-grade fever, conjunctivitis, and arthralgia, and an increase in rates of Guillain-Barre syndrome have been observed during ZIKV outbreaks (5). In Brazil, clusters of cases of acute exanthematous illness have been reported from various regions since late 2014, and in April 2015, ZIKV was identified as the etiologic agent (6). In May 2015, the Brazilian Ministry of Health recognized circulation of ZIKV in Brazil. We report epidemiologic findings for an ongoing outbreak of acute exanthematous illness in the population of Salvador, the third largest city in Brazil. The Salvador Epidemiologic Surveillance Office (ESO) was first alerted to cases of an acute exanthematous illness early in 2015. Reporting of cases increased during March, and in April the ESO established 10 public emergency health centers in Salvador as sentinel units for systematic surveillance of patients with acute exanthematous illness of unknown cause. The units searched retrospectively for suspected cases by review of medical charts of patients treated since February 15, continued with prospective case detection, and submitted weekly reports of identified cases to the ESO. During February 15−June 25, a total of 14,835 cases of an indeterminate acute exanthematous illness were reported from the 12 sanitary districts in Salvador. The overall attack rate was 5.5 cases/1,000 persons (4.6 cases/1,000 men and 6.3 cases/1,000 women, 8.2 cases/1,000 children 40 years of age). The epidemic curve peaked in the first week of May, which was 1 week after molecular diagnosis of ZIKV in 8 patients residing ≈50 km from Salvador and during a period of intense media coverage of the outbreak (Figure) (6). Reporting of suspected dengue cases in Salvador did not vary substantially from that in other years and was >5 times lower: 2,630 cases, of which 165/366 (45.1%) were positive for dengue IgM, 20/590 (3.4%) positive for dengue virus nonstructural protein 1, and 1/11 (9.1%) positive for dengue virus by reverse transcription PCR (Figure). During the same period, 58 cases of suspected chikungunya were reported and 24 patients with suspected Guillain-Barre syndrome were hospitalized. Figure Reported cases of indeterminate acute exanthematous illness and suspected dengue fever in Salvador, Brazil, by date of medical care, February 15−June 25, 2015. Letters indicate specific events. A) February 15: systematic reporting of cases of ... The median age of case-patients was 26 years (interquartile range 11–39 years), but all age groups were affected, which is a pattern typical of spread of new microorganisms (or subtypes) in a susceptible population. Median duration of symptoms at time of medical attention was 1 day (interquartile range 0–3 days). All patients had exanthema and most (12,711/14,093 [90.2%]) had pruritus. Fever (4,841/13,786, 35.1%), arthralgia (278/1,048 [26.5%]), headache (3,446/13,503 [25.6%]), and myalgia (223/1,033 [21.6%]) were less common. Serum samples from some patients were examined for rubella IgM (2/200, 1.0% positive), rubella IgG (15/18, 83.3% positive), measles IgM (0/11, 0% positive), dengue nonstructural protein 1 (3/185, 1.6% positive), dengue IgM (17/80, 21.3% positive), parvovirus B19 IgM (0/1, 0% positive), and parvovirus B19 IgG (1/1, 100% positive). Reverse transcription PCR was performed on 58 serum samples stored at −20°C and confirmed ZIKV in 3 (5.2%) samples, CHIKV in 3 (5.2%) samples, DENV type 3 in 1 (1.7%) sample, and DENV type 4 in 1 (1.7%) sample. Identification of ZIKV, CHIKV and DENV as etiologic agents of acute exanthematous illness suggests that these 3 Aedes spp. mosquito−transmitted viruses were co-circulating in Salvador and highlights the challenge in clinically differentiating these infections during outbreaks. Although we were not able to determine the specific incidence of each virus, the low frequency of fever and arthralgia, which are indicators of dengue and chikungunya, point to ZIKV as the probable cause of several of the reported cases. Furthermore, laboratory-confirmed cases of infection with ZIKV were simultaneously identified in other cities within metropolitan Salvador (6,7) and in other states in Brazil (8). Low diagnosis of ZIKV infection is likely because viremia levels among infected patients appear to be low (9). The spread of ZIKV represents an additional challenge for public health systems, particularly because of the risk for concurrent transmission of DENV and CHIKV by the same vectors, Ae. aegypti and Ae. albopictus mosquitoes, which are abundant throughout tropical and subtropical regions. To date, the largest outbreak of chikungunya in Brazil occurred in 2014 in Feira de Santana, Bahia, ≈100 km from Salvador, where dengue is also prevalent (10). This report illustrates the potential for explosive simultaneous outbreaks of ZIKV, CHIKV, and DENV in the Western Hemisphere and the increasing public health effects of Aedes spp. mosquitoes as vectors. The apparent increase in reports of Guillain-Barre syndrome during the outbreak deserves further investigation to elucidate whether this syndrome is associated with ZIKV infection. Public health authorities in Brazil and neighboring countries should plan accordingly.

Epidemiological surveillance for Human Bocavirus (HBoV) was conducted on 105 fecal specimens from children with acute gastroenteritis in Bahia, Brazil. Among of a total 105 stool samples, 44 samples were positive for HBoV as detected by nested-PCR. Of the 44 positive samples, co-infections with other enteric viruses (Norovirus, Adenovirus, and Rotavirus) were found in 12 pediatric patients. Mixed infections among HBoV with Norovirus were frequently observed in this population. The phylogenetic analysis identified the presence of HBoV-1, and HBoV 2A species. This study shows that HBoV is another viral pathogen in the etiology of acute gastroenteritis in children in Bahia, Brazil.

National Council for Scientific and Technological Development, Brazil (CNPq); Coordination for the Improvement of Higher Education Personnel, Ministry of Education, Brazil (CAPES)

Brazil is an endemic area for Dengue virus (DENV) infections. Recently, the country was affected by an outbreak caused by Chikungunya virus, and in the last year, a third arbovirus (Zika virus – ZIKV), caused a large number of infections.1 ZIKV is transmitted by mosquitoes, and was first isolated from a rhesus macaque placed as sentinel during a study about yellow fever in the Zika Forest, Uganda, Africa in 1947.2 Since that time, sporadic viral isolations from humans and from a variety of mosquitoes Aedes sp have been reported in Africa and Asia. ZIKV, a positive-sense, single-stranded RNA virus, member of genus Flavivirus, family Flaviviridae, is another flavivirus of a public health importance. The virus has been circulated in Southeast Asia for at least the past 50 years, and a recent epidemic on Yap Island, Federated States of Micronesia, demonstrated that ZIKV is also capable of causing human disease and is expanding its geographic distribution.3 In Bahia, in the first months of 2015, patients at Santa Helena Hospital in Camacari, a city distant 50 km from Salvador, the third largest city in Brazil, were given a presumptive diagnosis of an acute viral illness by emergency department physicians. The illness was characterized by maculopapular rash, fever, myalgias/arthralgia, conjunctivitis, severe rash in arms and legs, and low grade fever. This atypic, dengue-like illness was investigated to confirm the presence of several arbovirus by conventional RT-PCR: West Nile, Mayaro virus, Sant Louis virus and DENV. Since all tests resulted negative, there was only one more possibility to investigate: ZIKV. On March 26 2015, Campos et al. identified ZIKV in sera of affected patients, demonstrating, for the first time, circulation of ZIKV in Brazil and Latin America.4 It was immediately informed to the national health authorities (Ministry of Health, Brazil) and the presence of ZIKV (autoctones cases) in Bahia was

SUMMARYDengue virus, commonly transmitted by mosquitoes, causes a human disease of significant social impact and presents a serious public health problem in Brazil. This report describes the unusual emergence of DENV-4 in northern Brazil after a nearly 30-year-long absence. DENV-4 genotype I is of Asian origin and was identified in the serum of patients receiving treatment at a hospital serving the Salvador area (Brazilian state of Bahia). The identification of dengue virus serotypes through molecular and phylogenetic analysis is essential for predicting disease severity or fatal illness, principally in endemic countries such as Brazil.

Background: Yellow fever virus (YFV) is an RNA virus maintained in an enzootic, sylvatic cycle involving nonhuman primates (NHPs) and sylvatic mosquito vectors primarily of the genus Haemagogus and Sabethes. Transmission occasionally spills over to humans entering forested regions. In the Americas, urban transmission of YFV to humans has not occurred since the mid-1900s because of vaccination and near-elimination of the anthropophilic Aedes aegypti, the urban vector (1). However, concerns about reemergence of urban YFV have recently increased because of the reappearance and rapid spread of A aegypti in the urban environment. Furthermore, immunization coverage for YFV is insufficient because vaccination is generally indicated only for higher-risk populations, such as those living in or travelling to areas with sylvatic transmission. Fil: Paploski, Igor Adolfo Dexheimer. Universidade Federal da Bahia; Brasil Fil: Souza, Raquel Lima. Fundación Oswaldo Cruz; Brasil Fil: Tauro, Laura Beatriz. Fundación Oswaldo Cruz; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú | Universidad Nacional de Misiones. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú; Argentina Fil: Cardoso, Cristiane Wanderley. Secretaria Municipal de Saúde de Salvador; Brasil Fil: Mugabe, Vánio André. Universidade Federal da Bahia; Brasil Fil: Alves, Anna Beatriz Pereira Simões. Fundación Oswaldo Cruz; Brasil Fil: De Jesus Gomes, Joice. Fundación Oswaldo Cruz; Brasil Fil: Kikuti, Mariana. Universidade Federal da Bahia; Brasil Fil: Campos, Gubio Soares. Universidade Federal da Bahia; Brasil Fil: Sardi, Sílvia. Universidade Federal da Bahia; Brasil Fil: Weaver, Scott C.. University of Texas Medical Branch; Estados Unidos Fil: Reis, Mitermayer Galvão. Universidade Federal da Bahia; Brasil Fil: Kitron, Uriel D.. University of Emory; Estados Unidos Fil: Ribeiro, Guilherme Sousa. Universidade Federal da Bahia; Brasil

Dengue fever (DF) is a mosquito-borne viral disease of great concern in tropical and subtropical regions of the world. One important cause of the increase in DF is rapid development and urbanization has led to proliferation of the Aedes aegypti mosquito, the vector responsible for transmission of the illness. Surveillance of dengue virus (DENV) infection in Brazil shows the predominance of DENV-1, DENV-2, and DENV-3 until 2010. This study reports the reappearance of DENV-4 in Brazil for the first time in 30 years. Serum samples were collected from individuals (n = 214) exhibiting fever and muscular pain in Bahia, Brazil, during 2011-2012. These samples were subjected to reverse transcription-polymerase chain reaction (RT-PCR)/nested PCR, which revealed that 82% of samples were positive for DENV-4; most were older age groups and exhibited a serological pattern consistent with a primary infection. The cocirculation of multiple DENV serotypes within the same city places the population at risk for a fatal form of the disease. Therefore, with the increasing incidence of severe DF cases, early diagnosis will be a priority for public health efforts in Brazil.

Texto completo. Acesso restrito. p. 352–356 Submitted by Santiago Fabio (fabio.ssantiago@hotmail.com) on 2013-07-04T17:06:44Z No. of bitstreams: 1 2222222222222.pdf: 424706 bytes, checksum: 944826904f088f49e3251507838d5269 (MD5) Approved for entry into archive by Flávia Ferreira(flaviaccf@yahoo.com.br) on 2013-07-31T20:09:26Z (GMT) No. of bitstreams: 1 2222222222222.pdf: 424706 bytes, checksum: 944826904f088f49e3251507838d5269 (MD5) Made available in DSpace on 2013-07-31T20:09:26Z (GMT). No. of bitstreams: 1 2222222222222.pdf: 424706 bytes, checksum: 944826904f088f49e3251507838d5269 (MD5) Previous issue date: 2013 The genome of the Caprine Arthritis-Encephalitis Virus (CAEV) encodes the polycistronic precursor protein p55gag. Proteolytic cleavage of p55gag generates the viral core proteins. Some studies suggest that the CAEV p55gag protein contains epitopes or antigenic determinants for these core proteins. This work reinforces this hypothesis and demonstrates that monoclonal antibodies (MAbs) that are directed against the capsid protein (p28) of CAEV are also reactive against the precursor p55gag protein and the intermediate cleavage products, p44, p36 and p22. The major activity of the MAbs was directed against p28. The MAbF12 binding site in p28 was found to be a linear epitope with a structure that is stable after SDS treatment and remains unaltered after -mercaptoethanol ( -ME) treatment. The MAbF12 binding site in the p55gag, p36 and p22 proteins was found to be a linear epitope with cross-linked sulphide bonds. In conclusion, these findings suggest that the p28 epitope is presented differently from the epitope in the polycistronic precursor protein p55gag. The highly immunogenic p28 contains a linear epitope that is detergent-stable and is not altered by -ME treatment, whereas the p55gag epitope contains a linear epitope susceptible to denaturing agents. Salvador

Many established protocols are available to produce antigens for ELISA tests but expensive equipments, the purification methods or recombinant technologies are not always available to veterinary diagnostic laboratories. The aim of this work was to develop an alternative ELISA test to detect antibodies against Lentivirus Small Ruminant Virus (Caprine arthritis-encephalitis virus-CAEV) in goats, simple to elaborate with a low cost of production. The antigen was obtain from a whole cellular lysates of goat synovial membrane cells (GSMC) infected with CAEV and treated with SDS 0,1%. The whole-viral antigen was capable of detecting antibodies against several viral proteins including the p28 kDa (viral capsid), the p33 kDa, the p44 kDa (capsid precursors) and p97 kDa (transmembrane glycoprotein). The comparison with others assays as indirect immunofluorescence showed a high correlation with the ELISA results and the polimerasse chain reaction (PCR) demonstrated that most of the goat seropositives by ELISA, had proviral DNA in the milk cells. In conclusion, the viral antigen obtained from GSMC was simple to elaborate, low cost of production. The ELISA test was capable of detecting antibodies against CAEV, to detect early infection.

The oral route of human papillomavirus (HPV) transmission is not fully understood. It has been suggested that genital infection can act as a reservoir for oral HPV infection. We investigated the presence of oral HPV DNA and anti-HPV IgA in the buccal cavity of patients with a histopathologic diagnosis of cervical HPV infection. One hundred women underwent oral clinical examinations to detect HPV-DNA by polymerase chain reaction and salivary anti-HPV IgA by indirect immunofluorescence. Information on the personal habits of all the women was collected in personal interviews. Our results showed that 99% of the patients had no clinical manifestations of oral HPV. However, HPV DNA was detected in 81% of oral mucosa samples, and anti-HPV IgA was detected in the saliva of 44% of the patients. Consumption of alcoholic beverages was significantly associated with detection of oral HPV DNA and salivary anti-HPV IgA. Other behavioral risk factors associated with oral HPV and anti-HPV IgA are also discussed. In conclusion, patients with genital HPV infection are at risk for subclinical oral HPV infection. Thus, a molecular assay might be necessary to diagnose such infections.

Submitted by ROBERTO PAULO CORREIA DE ARAÚJO (ppgorgsistem@ufba.br) on 2017-06-05T17:06:13Z No. of bitstreams: 1 7_v.9_3.pdf: 625337 bytes, checksum: b9e70e51bd062c3e44b154a819253814 (MD5) Made available in DSpace on 2017-06-05T17:06:13Z (GMT). No. of bitstreams: 1 7_v.9_3.pdf: 625337 bytes, checksum: b9e70e51bd062c3e44b154a819253814 (MD5) Previous issue date: 2010-09 A infecção pelo vírus da dengue possui um amplo espectro de manifestações clínicas: desde sintomas semelhantes aos do resfriado comum até uma apresentação grave, podendo evoluir para óbito. Alguns dos quadros mais graves podem associarse à síndrome do choque da dengue, frequentemente, fatal em crianças. Essa infecção pode também determinar comprometimento da função hepática, com um quadro difuso de hepatite. Apesar de o acometimento hepático ser, geralmente autolimitado, alguns casos podem evoluir com maior gravidade. Foram avaliadas informações clínicas e laboratoriais relacionadas à função hepática em 55 pacientes pediátricos atendidos em um serviço de pronto atendimento em Salvador, que tiveram diagnóstico de dengue confirmado por quadro clínico e laboratorial (ELISA, imunocromatografia). Foram observados: 61,80% (34/55) de alteração na ALT e 50,90% (28/55) na AST. A variação dos valores de ALT foi de 7-432U/ l e de AST foi de 25-537U/l. Náuseas e (ou) vômitos foram referidos por 40,0% (22/55) dos pacientes, assim como dor abdominal, e 3(5,4%) apresentaram hepatomegalia. A elevação de enzimas hepáticas ocorreu em todos os pacientes diagnosticados com síndrome do choque da dengue, enquanto que a alteração de função hepática foi documentada em 51,0% dos pacientes diagnosticados com febre clássica da dengue. Esses dados revelam a importância da avaliação hepática sistemática nos pacientes com dengue clássico e dengue hemorrágica e a necessidade do treinamento dos profissionais de saúde para a atenção a esse aspecto na assistência de pacientes com essa infecção. Salvador

The molecular epidemiology of a recent norovirus (NoV) outbreak in Brazil performed by comparative analysis with Genebank NoV sequences showed that the GII.4 strain was responsible for 72.5% of all NoV-positive cases (58/80). Other detected NoV strains included GII.3 (7/80; 8.8%) and GII.9 (8/80; 10%). This is the first outbreak reported in Bahia state, Brazil, during June-July of 2006, where NoV was identified as the principal etiologic agent in hospitalized young adults with acute gastroenteritis symptoms. These findings suggest that GII.4 is a predominant circulating genotype in NoV outbreaks in Brazil.

Rotavirus is a major cause of infectious diarrhea in infants and young children. The objective of this study was to characterize the genotypes of Human Rotavirus found in children hospitalized with acute diarrhea in the Pediatric Hospital Prof. Hosannah de Oliveira of the UFBA in Salvador, Bahia, Brazil, during the years of 1999, 2000 and 2002. Fecal samples were analyzed (n=358) by methods EIARA and SDS-PAGE for detection of Rotavirus. Positive samples of one or two of these methods (n=168) were submitted to RT-PCR and Multiplex-Nested PCR to determine genotypes G and P. A hundred sixty-eight (46.9%) samples were positive and 190 (53.1%) negative. Only 17 (4.7%) samples had divergent results. The distribution of genotypes G during the first year, showed that the genotype G9 was present in 96,8% of the analyzed samples, in the second year, it was responsible for 96% and in the third year, 88,1%. The characterization of genotypes P demonstrated that the genotype P1A[8] was the most outstanding in all years. In this study we discuss the benefit to control the genotypes of Rotavirus through the molecular characterization for the development of potential vaccines.

p.124-131, Artigo original Submitted by ROBERTO PAULO CORREIA DE ARAÚJO (ppgorgsistem@ufba.br) on 2016-09-14T14:41:51Z No. of bitstreams: 1 4_v.5_2.pdf: 62550 bytes, checksum: 5464aa4379b76ed886afcb388b4b9fe7 (MD5) Made available in DSpace on 2016-09-14T14:41:51Z (GMT). No. of bitstreams: 1 4_v.5_2.pdf: 62550 bytes, checksum: 5464aa4379b76ed886afcb388b4b9fe7 (MD5) Previous issue date: 2006-05 O vírus da Artrite encefalite caprina é o agente causal de uma doença progressiva e debilitante em caprinos, em que as células da linhagem monócito/macrófago são as principais hospedeiras do vírus in vivo. Essas células estão presentes no colostro, no leite ou no sangue. Estudos demonstram que a replicação viral é dependente do nível de maturação ou diferenciação da célula monocítica, influenciando na sensibilidade de detecção do vírus. Neste estudo, utilizamos o cocultivo de monócitos/macrófagos com células de membrana sinovial de cabras, para o isolamento do vírus circulante no campo; a técnica de double nested PCR (dn PCR) dos co-cultivos e sangue, viabilizou a confirmação do isolamento viral, e a detecção direta do vírus no sangue. Através dessa técnica, detectou-se a presença do DNA proviral em animais soronegativos por Imunodifusão em Gel de Agarose (IDGA); esses achados confirmam que o tempo entre o inicio da infecção e o aparecimento de anticorpos no sangue é variável, facilitando a permanência de animais falsos-negativos no rebanho. Nas amostras processadas, tivemos uma divergência de resultados entre amostras sorologicamente positivas, entretanto, negativas por dn PCR, quando se utilizou a amplificação direta do sangue. Quanto ao cultivo de monócitos/ macrófagos in vitro e posterior co-cultivo com células de MSC obteve-se êxito pelo isolamento do vírus em quatro animais, havendo sido um deles soronegativo. O presente estudo demonstra o primeiro isolamento do vírus nos rebanhos do estado da Bahia, além da implementação da técnica de dn PCR, em co-cultivo de células. Salvador

Texto completo: acesso restrito. p. 196–203 Submitted by Edileide Reis (leyde-landy@hotmail.com) on 2013-08-27T15:03:28Z No. of bitstreams: 1 1-s2.0-S1046592805003359-main.pdf: 1116869 bytes, checksum: 9868a3eaec7c83a1a863781bd5c20e9f (MD5) Approved for entry into archive by Rodrigo Meirelles (rodrigomei@ufba.br) on 2013-11-19T20:49:41Z (GMT) No. of bitstreams: 1 1-s2.0-S1046592805003359-main.pdf: 1116869 bytes, checksum: 9868a3eaec7c83a1a863781bd5c20e9f (MD5) Made available in DSpace on 2013-11-19T20:49:41Z (GMT). No. of bitstreams: 1 1-s2.0-S1046592805003359-main.pdf: 1116869 bytes, checksum: 9868a3eaec7c83a1a863781bd5c20e9f (MD5) Previous issue date: 2006 Rotavirus VP8* subunit is the minor trypsin cleavage product of the spike protein VP4, which is the major determinant of the viral infectivity and neutralization. To study the structure–function relationship of this fragment and to obtain type-specific reagents, substantial amounts of this protein are needed. Thus, full-length VP8* cDNA, including the entire trypsin cleavage-encoding region in gene 4, was synthesized and amplified by RT-PCR from total RNA purified from bovine rotavirus strain C486 propagated in MA104 cell culture. The extended VP8* cDNA (VP8ext) was cloned into the pGEM-T Easy plasmid and subcloned into the Escherichia coli expression plasmid pET28a(+). The correspondent 30 kDa protein was overexpressed in E. coli BL21(DE3)pLysS cells under the control of the T7 promoter. The identity and the antigenicity of VP8ext were confirmed on Western blots using anti-His and anti-rotavirus antibodies. Immobilized Ni-ion affinity chromatography was used to purify the expressed protein resulting in a yield of 4 mg of VP8ext per liter of induced E. coli culture. Our results indicate that VP8ext maintained its native antigenicity and specificity, providing a good source of antigen for the production of P type-specific immune reagents. Detailed structural analysis of pure recombinant VP8 subunit should allow a better understanding of its role in cell attachment and rotavirus tropism. Application of similar procedure to distinct rotavirus P serotypes should provide valuable P serotype-specific immune reagents for rotavirus diagnostics and epidemiologic surveys.

Two hundred eight of 648 (32%) diarrheal stool samples collected from hospitalized children under 5 years of age during a 3-year period (1999, 2000, and 2002) in the city of Salvador, in the state of Bahia, Brazil, were rotavirus positive. One hundred sixty-four of 208 (78.8%) rotavirus-positive samples had genotype G9 specificity, predominantly in association with P[8]. Other specificities detected were G1 (12.0%) and G4 (1.4%). Viruses with G2, G3, or P[4] specificity were not detected. Rotavirus genotype G9 predominated during each of the three seasons studied; it represented 89.2% of rotavirus strains detected in 1999, 85.3% in 2000, and 74.5% in 2002. G1 viruses (the globally most common G type) have a unique epidemiological characteristic of maintaining predominance during multiple consecutive rotavirus seasons. We have shown in this study for the first time that the G9 viruses also have a similar epidemiological characteristic, albeit for a shorter period of surveillance. The next generation of rotavirus vaccines will need to provide adequate protection against disease caused by G9 viruses.