Marcos César Lima de Mendonça

Guillain-Barré syndrome associated with Zika virus infection

A 24-year-old housekeeper presented to hospital in Rio de Janeiro in June, 2014, with headache, fever, and a rash, 5 days after waking with a severe generalised headache, retro-orbital pain, weakness, and paraesthesia of the hands and feet. 2 days later she developed fever (axillary temperature 42°C), chills, and a pruritic rash on the face, abdomen, chest, and arms. By day 4, she was afebrile but had painful swelling of the hands (appendix) and feet, diffi culty walking, and disseminated rash. She had had dengue 5 years previously, had not travelled recently, and did not recall any tick or mosquito bites. On examination, she was alert and fully oriented. Axillary temperature was 36·7°C, pulse 90 beats per min, blood pressure 100/60 mm Hg, and respiratory rate 20 breaths per min. She had a diff use erythematous macular rash, bilateral non-purulent conjunctival hyperaemia, enanthema of the palate, one enlarged painless cervical lymph node, and swelling of the hands and feet, but no signs of meningism. She had reduced strength in the legs, absent deep tendon refl exes at the knees and ankles, and both plantars were absent; sensation to light touch was reduced in the legs, but she had no urinary retention or ataxia. Examination, including neurological examination of the arms, was otherwise normal. Lumbar puncture (day 6), nerve conduction studies and an electromyogram (day 10), and a non-enhanced MRI (day 13) were normal. From day 10 the rash and swelling began to resolve with supportive treatment. By day 13 she was fully mobile and could be discharged. At follow-up on day 41, her only remaining symptom was persistent headache. We investigated her serum and cerebrospinal fl uid (CSF) for dengue, chikungunya, and Zika viruses. Realtime PCR for dengue and chikungunya was negative, but PCR was positive for Zika virus in serum (day 5), CSF (day 6), saliva (day 10), and urine (day 11). The CSF and acute and convalescent serum were negative for dengue and chikungunya by IgM-capture ELISA. Zika ELISA was not available. To identify the Zika virus genotype we sequenced 327 base pair amplicons encompassing the envelope protein, and identifi ed the Asian lineage of Zika in the CSF (fi gure). Like dengue and chikungunya, Zika virus causes a febrile illness with rash. During the 2013 outbreak of Zika virus in French Polynesia an apparent increase in Guillain-Barre syndrome incidence was noted but with no baseline data for comparison. One case had antibodies against Zika and dengue viruses (which can also trigger Guillain-Barre syndrome), but no virus was detected. Our patient had no evidence of dengue or chikungunya infection, but Zika was found in the CSF by PCR, and unusually she also had high grade fever and clinical features consistent with paraparetic Guillain-Barre syndrome, a rare atypical presentation. CSF and neurophysiological investigations were normal, as is often found early in Guillain-Barre syndrome. She met Level III of diagnostic certainty for Guillain-Barre syndrome in the Brighton classifi cation (consistent clinical features, but no supporting CSF or neurophysiology evidence). Our case highlights the potential for neurotropism of Zika virus, and the need to consider this emerging virus as a mosquito-borne cause of fever, rash, and neurological disease.

First detection of autochthonous Zika virus transmission in a HIV-infected patient in Rio de Janeiro, Brazil

Since May 2015, Brazils Ministry of Health has reported autochthonous transmission of Zika virus (ZIKV) in some states of the country. Simultaneous circulation of Dengue, Chikungunya and ZIKV in the country hinder both the diagnosis and the therapeutic approach of patients seeking care with acute febrile illnesses especially in patients with comorbidities. The association between HIV infection and endemic diseases has been described especially in tropical regions with varying levels of complications, although there has been no report of ZIKV in HIV-infected patients. We report the first autochthonous case of laboratory confirmed ZIKV infection in a HIV-infected patient in Rio de Janeiro, Brazil. He evolved with only mild symptoms and recovered well without major laboratory abnormalities. Phylogenetic analysis of the ZIKV detected in the patient sera clustered within the Asian clade. To the best of our knowledge, this is the first time that Zika virus co-infection is reported in a HIV-infected patient.

Genetic diversity of porcine enteric caliciviruses in pigs raised in Rio de Janeiro State, Brazil

Porcine enteric caliciviruses (PEC) belong to the genera Norovirus and Sapovirus within the family Caliciviridae. They are enteric pathogens and are considered potential zoonotic agents. In this study, the circulation of PEC was evaluated by RT-PCR of stool samples and intestinal contents of pigs raised in Rio de Janeiro State, Brazil. Both porcine norovirus (PoNoV) and porcine sapovirus (PoSaV) were detected. The PoNoV strains were classified as genogroup II, genotypes 11, 18 and 19. The PoSaV strains were classified as genogroups III and VII, though some strains could not be classified into any established genogroup, potentially representing a new one. PEC were detected mainly in animals without clinical signs of gastroenteritis.

Phylogenetic analysis of human P[8]G9 rotavirus strains circulating in Brazil reveals the presence of a novel genetic variant

BACKGROUND Group A rotavirus (RV-A) genotype P[8]G9 has emerged as one of the leading causes of gastroenteritis in children worldwide and currently is recognized as one of the five most common genotypes detected in humans. High intragenotype diversity in G9 RV-A has been observed, and nowadays, based on the genetic variability of the VP7 gene, six different phylogenetic lineages and eleven sublineages were described. OBJECTIVES To study the degree of genetic variation and evolution of Brazilian P[8]G9 RV-A strains. STUDY DESIGN Phylogenetic analysis of 19 P[8]G9 RV-A strains isolated from 2004 to 2007 in five different Brazilian states was conducted using the NSP1, NSP3, NSP5, VP4 and VP7 genes. For the VP4 and VP7 genes, 3D protein structure predictions were generated to analyze the spatial distribution of amino acid substitutions observed in Brazilian strains. RESULTS Based on the phylogenetic analyses, all Brazilian strains clustered within lineage G9-III and P[8]-3 for VP7 and VP4, respectively, and were classified as genotype A1, T1 and H1 for the NSP1, NSP3 and NSP5 genes, respectively. Interestingly, all the strains isolated in Acre State (Northern Brazil) formed a closely related cluster clearly separated from the other Brazilian and prototype strains with regard to the five genes studied. Unique amino acid substitutions were observed in Acre strains in comparison with the prototype and Brazilian strains. CONCLUSION Inclusion of Acre strains in the phylogenetic analysis revealed the presence of a novel genetic variant and demonstrated a diversification of P[8]G9 rotaviruses in Brazil.

VP7 Gene of Human Rotavirus A Genotype G5: Phylogenetic Analysis Reveals the Existence of Three Different Lineages Worldwide

Group A rotavirus (RV‐A) genotype G5, which is common in pigs, was also detected in children with severe diarrhea in Brazil, Argentina, Paraguay, Cameroon, China, Thailand, and Vietnam. To evaluate the evolutionary relationship among RV‐A G5 strains, the VP7 and VP4 genes of 28 Brazilian RV‐A G5 human strains, sampled between 1986 and 2005, were sequenced and compared with other RV‐A G5 strains currently circulating worldwide in animals and humans. The phylogenetic analysis of RV‐A G5 VP7 gene strains demonstrates the existence of three main lineages: (a) Lineage I: Brazilian strains grouped with three porcine strains from Thailand; (b) Lineage II: porcine, bovine, and equine strains from different regions; (c) Lineage III: human strains isolated in Asia and Africa, and two porcine strains from Argentina. The VP8* (*non‐typable) subunit of VP4 gene sequencing showed that all P[8] strains fell into three major genetic lineages: P[8]‐1; P[8]‐2; and P[8]‐3. These results showed that the RV‐A G5 strains circulating in humans are the result of two independent zoonotic transmission events, most likely from pigs. J. Med. Virol. 83:357–366, 2011.

Phylogenetic analysis of VP1, VP2, and VP3 gene segments of genotype G5 group A rotavirus strains circulating in Brazil between 1986 and 2005

Genotype G5 group A rotavirus (RV-A), which is common in pigs and also detected in horses and cattle, circulated as endemic genotype in the 1980s and early 1990s in Brazil. After 1996, G5 RV-A has been replaced by G9 RV-A, becoming only sporadically detected. Recently, G5 has been reported in children with severe diarrhea in Argentina, Cameroon, Paraguay, Peoples Republic of China, and Vietnam, suggesting that, although uncommon in humans, it has a worldwide distribution. In a previous study, Brazilian G5 RV-A VP7 gene analysis demonstrated the existence of three main lineages: I, II, and III; all Brazilian strains and three porcine strains from Thailand grouped inside Lineage I. The VP8(*) subunit of VP4 gene showed that all P[8] strains fell into three major genetic lineages: P[8]-1; P[8]-2; and P[8]-3. Partial sequencing and phylogenetic analysis of VP1, VP2 and VP3 genes of P[8]G5 human RV-A strains were determined from 28 Brazilian strains collected from 1986 to 2005. The VP1-VP3 partial sequences analysis showed that the Brazilian strains have high amino acid identity with the human RV-A prototype IAL28 and other Wa-like genogroup strains. It was also shown that G5 RV-A Brazilian VP1-VP3 and VP7 sequences have a similar pattern of gouping: The study strains and the G5 prototype strain IAL-28 grouped together, while other prototypes, like OSU grouped separately. These results suggest that the core protein genes (VP1-VP3) of the G5 RV-A Brazilian strains might have originated from porcine and human strains. Phylogenetic analyses of VP7, VP4, VP1, VP2, and VP3 genes of P[8]G5 strains revealed a conserved genomic constellation (G5-P[8]-R1-C1-M1) with sequence similarity to Wa-like strains: IAL28, Wa, BE00048, CK00032, CK00033, DC4772 and DC1898, suggesting that despite the differences in genotypes (i.e., G5, G1 and G3) these viruses are genetically similar. The results presented here are fundamental to understand the epidemiology and evolution of genotype G5 RV-A and demonstrate the importance of continuous monitoring and molecular characterization of RV-A strains circulating in human and animal populations.

Dengue severity associated with age and a new lineage of dengue virus-type 2 during an outbreak in Rio De Janeiro, Brazil

Dengue virus‐type 2 (DENV‐2) caused three outbreaks, in the years 1990, 1998, and 2008, in Rio de Janeiro, Brazil. The 2008 outbreak was the most severe in reported cases, hospitalizations, and deaths. To investigate virological and epidemiological factors that may have contributed to the pathogenic profile of 2008 epidemic, 102 patients sera obtained during the epidemic and inter‐epidemic periods of three outbreaks were analysed by qRT‐PCR to estimate viremia levels and their correlation with the clinical, immunological, and demographic patient characteristics. DENV‐2 isolates from the outbreaks were sequenced. Two DENV‐2 lineages (I and II) of the American/Asian genotype were confirmed, each exclusive for 1990–2002 and 2007–2011, respectively. The mean viremia level in the 2008 samples was two orders of magnitude higher than that of the 1990–2002 samples. Severe dengue cases increased from 31% in 1990–2002 to 69% in 2007–2011; in patients aged ≤15 years, from 3% in 1990–2002 to 37% in 2007–2011. The DENV‐2 lineage II and younger age significantly contributed to the pathogenic profile of 2008 epidemic in Rio de Janeiro. J. Med. Virol. 88:1130–1136, 2016.

Phylogenetic analyses of chikungunya virus among travelers in Rio de Janeiro, Brazil, 2014-2015.

Chikungunya virus (CHIKV) is a mosquito-borne pathogen that emerged in Brazil by late 2014. In the country, two CHIKV foci characterized by the East/Central/South Africa and Asian genotypes, were established in North and Northeast regions. We characterized, by phylogenetic analyses of full and partial genomes, CHIKV from Rio de Janeiro state (2014-2015). These CHIKV strains belong to the Asian genotype, which is the determinant of the current Northern Brazilian focus, even though the genome sequence presents particular single nucleotide variations. This study provides the first genetic characterisation of CHIKV in Rio de Janeiro and highlights the potential impact of human mobility in the spread of an arthropod-borne virus.

Genotyping of human parvovirus B19 in clinical samples from Brazil and Paraguay using heteroduplex mobility assay, single-stranded conformation polymorphism and nucleotide sequencing

Heteroduplex mobility assay, single-stranded conformation polymorphism and nucleotide sequencing were utilised to genotype human parvovirus B19 samples from Brazil and Paraguay. Ninety-seven serum samples were collected from individuals presenting with abortion or erythema infectiosum, arthropathies, severe anaemia and transient aplastic crisis; two additional skin samples were collected by biopsy. After the procedure, all clinical samples were classified as genotype 1.

Parvovirus B19 1A complete genome from a fatal case in Brazil

Parvovirus B19 (B19V) infects individuals worldwide and is associated with an ample range of pathologies and clinical manifestations. B19V is classified into three distinct genotypes, all identified in Brazil. Here, we report a complete sequence of a B19V genotype 1A that was obtained by high-throughput metagenomic sequencing. This genome provides information that will contribute to the studies on B19V epidemiology and evolution.