High Prevalence and Onward Transmission of Non-Pandemic HIV-1 Subtype B Clades in Northern and Northeastern Brazilian Regions

<div><p>The Human immunodeficiency virus type-1 (HIV-1) epidemic in Brazil is mainly driven by the subtype B pandemic lineage (B_PANDEMIC), while Caribbean non-pandemic subtype B clades (B_CAR) seem to account for a very low fraction of HIV-infections in this country. The molecular characteristics of the HIV-1 subtype B strains disseminated in the Northern and Northeastern Brazilian regions, however, have not been explored so far. In this study, we estimate the prevalence of the HIV-1 B_PANDEMIC and B_CAR clades across different Brazilian regions and we reconstruct the spatiotemporal dynamics of dissemination of the major Brazilian B_CAR clades. A total of 2,682 HIV-1 subtype B <i>pol</i> sequences collected from 21 different Brazilian states from the five country regions between 1998 and 2013 were analyzed. Maximum Likelihood phylogenetic analyses revealed that the B_CAR strains reached 16 out 21 Brazilian states here analyzed. The B_CAR clades comprise a low fraction (<10%) of subtype B infections in most Brazilian states analyzed, with exception of Roraima (41%), Amazonas (14%) and Maranhão (14%). Bayesian phylogeographic analyses indicate that B_CAR strains originally from the Hispaniola and Trinidad and Tobago were introduced at multiple times into different states from all Brazilian regions and a few of those strains, probably introduced into Roraima, Maranhão and São Paulo between the late 1970s and the early 1980s, established secondary outbreaks in the Brazilian population. These results support that the HIV-1 subtype B epidemics in some Brazilian states from the Northern and Northeastern regions display a unique molecular pattern characterized by the high prevalence of B_CAR lineages, which probably reflects a strong epidemiological link with the HIV-1 epidemics in the Caribbean region.</p></div

List of mutations known to confer reduced susceptibility to antiretroviral agents among ART-experienced HIV-1-infected patients attended at the Public Health Central Laboratory in Boa Vista, Roraima.

<p>List of mutations known to confer reduced susceptibility to antiretroviral agents among ART-experienced HIV-1-infected patients attended at the Public Health Central Laboratory in Boa Vista, Roraima.</p

Spatiotemporal dynamics of dissemination of HIV-1 B_CAR clades circulating in the Caribbean and Brazil.

<p>A) Time-scaled Bayesian MCMC tree of <i>pol</i> PR/RT sequences of HIV-1 B_CAR lineages from Brazil (n = 97) and the Caribbean (<i>n</i> = 258), and subtype D reference sequences (<i>n</i> = 10) from the Democratic Republic of Congo. Branches are colored according to the most probable location state of their descendent nodes as indicated in the legend at right. Colored boxes indicate the positions of major B_CAR clades detected in Brazil, Jamaica and Trinidad and Tobago. Branch lengths are depicted in units of time (years). The tree was automatically rooted under the assumption of a relaxed molecular clock. B) Lines between locations represent branches in the Bayesian MCC tree along which location transitions occurred and were colored according to the location of origin (see the legend at left). Maps were created from templates obtained from d-maps.com (Caribbean: <a href="http://d-maps.com/carte.php?numcar=1389&lang=en" target="_blank">http://d-maps.com/carte.php?numcar=1389&lang=en</a>; South America: <a href="http://d-maps.com/carte.php?numcar=2313&lang=en" target="_blank">http://d-maps.com/carte.php?numcar=2313&lang=en</a>; and Brazil: <a href="http://d-maps.com/carte.php?numcar=4843&lang=en" target="_blank">http://d-maps.com/carte.php?numcar=4843&lang=en</a>). AC: Acre; AM: Amazonas; AP: Amapá; CD: Democratic Republic of Congo; ES: Espírito Santo; GO: Goiás; HISP: Hispaniola; JM: Jamaica; MA: Maranhão; MG: Minas Gerais; MS: Mato Grosso do Sul; PA: Pará; PI: Piauí; RJ: Rio de Janeiro; RR: Roraima; RS: Rio Grande do Sul; SP: Sao Paulo; TO: Tocantins; TT: Trinidad and Tobago.</p

Epidemiological and clinical characteristics of HIV-1-infected patients attended at the Public Health Central Laboratory in Boa Vista, Roraima.

<p>Epidemiological and clinical characteristics of HIV-1-infected patients attended at the Public Health Central Laboratory in Boa Vista, Roraima.</p

HIV-1 subtype B <i>pol</i> (PR/RT and RT) Brazilian sequences.

<p>HIV-1 subtype B <i>pol</i> (PR/RT and RT) Brazilian sequences.</p

Bayesian T_MRCA estimates for major B_CAR clades from Brazil and the Caribbean.

<p>Bayesian T_MRCA estimates for major B_CAR clades from Brazil and the Caribbean.</p

Estimated proportion of B_CAR and B_PANDEMIC clades among HIV-1 subtype B infected individuals from different Brazilian states.

<p>The total number of sequences analyzed in each locality is indicated. States were colored according to the Brazilian region of origin as indicated in the legend at bottom, with exception of those states with no HIV-1 sequences included in this study (white). Map was created from a template obtained from d-maps.com (<a href="http://d-maps.com/carte.php?numcar=4843&lang=en" target="_blank">http://d-maps.com/carte.php?numcar=4843&lang=en</a>).</p

First evidence of Zika virus venereal transmission in Aedes aegypti mosquitoes

<div><p> BACKGROUND Aedes aegypti is considered the main Zika virus (ZIKV) vector, and is thought to be responsible for the 2015-2016 outbreak in Brazil. Zika positive Ae. aegypti males collected in the field suggest that vertical and/or venereal transmission of ZIKV may occur. OBJECTIVES In this study, we aimed to demonstrate that venereal transmission of ZIKV by Ae. aegypti can occur under laboratory conditions. METHODS Ae. aegypti collected in the city of Manaus, confirmed as negative for Zika, Dengue and Chikungunya virus by reverse transcription real-time polymerase chain reaction (RT-qPCR) (AaM3V- strain), were reared under laboratory conditions and used for the experiments. The ZIKV used in this study was isolated from a patient presenting with symptoms; ZIKV was confirmed by RT-qPCR. Experiment 1: virgin male mosquitoes of AaM3V- strain were intrathoracically inoculated with a ZIKV suspension; four days after injection, they were transferred to a cage containing virgin females of AaM3V- strain and left to copulate for five days. Experiment 2: virgin female mosquitoes of AaM3V- strain were orally infected with a ZIKV suspension by blood feeding membrane assay; nine days after blood feeding, they were placed in cages with Ae. aegypti AaM3V- virgin males and left to copulate for four days. After copulation, all mosquitoes were individually evaluated for viral infection by RT-qPCR. FINDINGS The mean infection rate in Experiment 1 and Experiment 2 was 45% and 35%, respectively. In both experiments, cycle threshold values ranged from 13 to 35, indicating the presence of viral genomes. MAIN CONCLUSION Ae. aegypti males intrathoracically inoculated with a ZIKV suspension are infected and can transmit the virus to uninfected females by mating. Moreover, Ae. aegypti females orally infected with a ZIKV suspension can transmit the virus to uninfected males by copulation. This study shows that ZIKV infection of Ae. aegypti mosquitoes occurs not only during blood feeding, but also during copulation.</p></div

Table_1_Toll-Like Receptor-1 Single-Nucleotide Polymorphism 1805T/G Is Associated With Predisposition to Multibacillary Tuberculosis.docx

<p>Tuberculosis (TB), caused by mycobacterial species of the Mycobacterium tuberculosis complex, is a serious global health issue. Brazil is among the 22 countries with the highest number of TB cases, and the state of Amazonas has the highest incidence of TB cases in the country. Toll-like receptors (TLRs) are important pattern recognition receptors of the innate immunity and play a key role in orchestrating an effective immune response. We investigated whether the single-nucleotide polymorphisms (SNPs) 1805T/G TLR1, 2258G/A TLR2, 896A/G and 1196C/T of TLR4, 745T/C TLR6, and −1237A/G and −1486A/G of TLR9 are associated with the predisposition to TB and/or bacillary load. The SNPs genotyping was performed by nucleotide sequencing in 263 TB patients and 232 healthy controls residing in the state of Amazonas. Alleles and genotypes frequencies were similar between patients and healthy individuals for most of the investigated SNPs. Stratification of the TB patients according to their bacillary load showed that the genotype 1805TT TLR1 (rs5743618) was prevalent among paucibacillary patients [odds ratio (OR) = 0.38; 95% confidence interval (CI) = 0.19–0.76; p = 0.009] while the genotype 1805TG was common among multibacillary patients (OR = 3.72; CI = 1.65–8.4; p = 0.004). Comparison of demographic characteristics of patients to controls showed that TB is strongly associated with smoking (OR = 6.55; 95% CI = 3.2–13.6; p < 0.0001); alcohol use disorder (OR = 7.14; 95% CI = 3.7–13.9; p < 0.0001); and male gender (OR = 3.66; 95% CI = 2.52–5.3; p < 0.0001). Multivariate logistic regression demonstrated that alcoholism (OR = 2.93; 95% CI = 1.05–8.16; p = 0.03) and the 1805G allele (OR = 2.75; 95% CI = 1.33–5.7; p = 0.006) are predictive variables for multibacillary TB. Altogether, we suggest that the TLR1 1805G allele may be a relevant immunogenetic factor for the epidemiology of TB together with environmental, sociodemographic, and behavioral factors.</p

Patients Demographics, Clinical Characteristics and Distribution of Genital Ulcer-Causing Pathogens by M-PCR.

<p>HSV-1: herpes simplex virus type 1; HSV-2: herpes simplex virus type 2; TP: <i>T. pallidum</i>; HSV: herpes simplex virus type 1 and type 2; ND: pathogen not detected by M-PCR.</p