Human G9P[8] rotavirus strains circulating in Cameroon, 1999-2000: Genetic relationships with other G9 strains and detection of a new G9 subtype.

Group A rotaviruses (RV-A) are the leading cause of viral gastroenteritis in children worldwide and genotype G9P[8] is one of the five most common genotypes detected in humans. In order to gain insight into the degree of genetic variability of G9P[8] strains circulating in Cameroon, stool samples were collected during the 1999-2000 rotavirus season in two different geographic regions in Cameroon (Southwest and Western Regions). By RT-PCR, 15 G9P[8] strains (15/89=16.8%) were identified whose genomic configurations was subsequently determined by complete or partial gene sequencing. In general, all Cameroonian G9 strains clustered into current globally-spread sublineages of the VP7 gene and displayed 86.6-100% nucleotide identity amongst themselves and 81.2-99.5% nucleotide identity with global G9 strains. The full genome classification of all Cameroonian strains was G9-P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1 but phylogenetic analysis of each gene revealed that the strains were spread across 4 or more distinct lineages. An unusual strain, RVA/Human-wt/CMR/6788/1999/G9P[8], which shared the genomic constellation of other Cameroonian G9P[8] strains, contained a novel G9 subtype which diverged significantly (18.8% nucleotide and 19% amino acid distance) from previously described G9 strains. Nucleotide and amino acid alignments revealed that the 3' end of this gene is highly divergent from other G9 VP7 genes suggesting that it arose through extensive accumulation of point mutations. The results of this study demonstrate that diverse G9 strains circulated in Cameroon during 1999-2000.

Performance of human immunostimulating agents in the improvement of fish cytogenetic preparations.

Inoculation with bacterial or fungal antigens that stimulate cell proliferation has been widely used to obtain metaphases for cytogenetic studies of fish. We evaluated the potential of new pharmaceutical compounds as mitogenic agents in fish, testing the efficacy of Aminovac (mixed antigens and epsilon-acetamidocaproic acid), Broncho-Vaxom (bacterial lysate) and Estimoral (bacterial lysate) to increase the mitotic index in fingerlings of the Neotropical fish Prochilodus brevis (Prochilodontidae) and Hoplias malabaricus (Erythrinidae), which were obtained from an aquaculture facility. The animals were treated with intramuscular or intraperitoneal injections of 1 mL/50 g body weight of each compound. After 24 h, cytogenetic analyses were performed. All immunostimulants tested significantly stimulated cell division, although Aminovac proved to be the most efficient, leading to a 5-fold increase in the number of metaphase cells compared to the control group and to a 2-fold increase compared to conventional yeast inoculation. This compound facilitates fish cytogenetics analyses as it stimulates the proliferation of defense cells and reduces loss of samples. It will be especially useful for the study of specimens that either have a high commercial value or are fragile, small and/or rare.

Detection of uncommon rotavirus A strains P[8]G8 and P[4]G8 in the city of Rio de Janeiro, 2002.

Group A rotaviruses (RV-A) are the major cause of gastroenteritis in infants and young children around the world. Each year RV-A causes approximately 11 million episodes of severe diarrhea, with an estimated of 611,000 deaths. Epidemiologic surveys have identified P[8]G1, P[4]G2, P[8]G3, P[8]G4, and P[8]G9 as the most common global genotypes associated with diarrhea in children up to 5-year old. Surveillance studies and documentation of RV-A G and P genotypes is necessary for a comprehensive evaluation of the evolution of new strains, and assessing the capability of vaccines to provide heterotypic protection. It is known that reassortments are the driving force for genetic diversity through sudden changes in RV-A genome. In this study, we identified two unusual P/G combinations, P[8]G8 and P[4]G8, occurring in Rio de Janeiro during 2002. Results obtained in this study suggest that P[8]G8 RV-A strain originated from a reassortment event that occurred between RV-A P[4]G8 and P[8]G9 strains circulating in Rio de Janeiro in the same year. G8 strains identified in this study, as well as G8 strains detected in Recife by Montenegro et al. [Montenegro et al. (2007) J Med Virol 79: 335-340], showed a close genetic relationship with strains from Africa, where this genotype have become prevalent.

Detection of caliciviruses associated with acute infantile gastroenteritis in Salvador, an urban center in Northeast Brazil

Acute gastroenteritis caused by viruses is one of the leading causes of infantile morbidity. The aim of the present study was to investigate the presence of human caliciviruses of the genera norovirus and sapovirus in children up to 3 years of age with acute gastroenteritis from low-income communities in the city of Salvador, Brazil. This study is an extension of previous work carried out to establish the profile of the most prevalent enteric pathogens present in these communities. In this report, 139 fecal samples, collected from July 2001 to January 2002 were analyzed by RT-PCR and 13 (9 percent) were positive for human caliciviruses. By sequencing, seven isolates were characterized as norovirus genogroup GII and one as sapovirus genotype GII/1. Sequencing of the previously detected group-A rotaviruses and human astroviruses was also performed and revealed the circulation of rotavirus group A genotypes G1P[8] and G9P[8], and human astrovirus genotypes 6, 7, and 8. No mixed infection was observed. Community-based studies provide geographically representative information on disease burden. However, there are only a few reports in developing countries concerning the genotypes of the most important gastroenteric viruses detected in such communities. The present findings demonstrate the wide diversity of genotypes of the most important viruses responsible for acute gastroenteritis circulating in low-income communities.

Detection of caliciviruses associated with acute infantile gastroenteritis in Salvador, an urban center in Northeast Brazil.

Acute gastroenteritis caused by viruses is one of the leading causes of infantile morbidity. The aim of the present study was to investigate the presence of human caliciviruses of the genera norovirus and sapovirus in children up to 3 years of age with acute gastroenteritis from low-income communities in the city of Salvador, Brazil. This study is an extension of previous work carried out to establish the profile of the most prevalent enteric pathogens present in these communities. In this report, 139 fecal samples, collected from July 2001 to January 2002 were analyzed by RT-PCR and 13 (9%) were positive for human caliciviruses. By sequencing, seven isolates were characterized as norovirus genogroup GII and one as sapovirus genotype GII/1. Sequencing of the previously detected group-A rotaviruses and human astroviruses was also performed and revealed the circulation of rotavirus group A genotypes G1P[8] and G9P[8], and human astrovirus genotypes 6, 7, and 8. No mixed infection was observed. Community-based studies provide geographically representative information on disease burden. However, there are only a few reports in developing countries concerning the genotypes of the most important gastroenteric viruses detected in such communities. The present findings demonstrate the wide diversity of genotypes of the most important viruses responsible for acute gastroenteritis circulating in low-income communities.