Prospective hospital-based surveillance to estimate rotavirus disease burden in the Gauteng and North West Province of South Africa during 2003-2005.

BACKGROUND: Rotavirus is considered to be the most common cause of serious acute dehydrating diarrhea worldwide. However, there is a scarcity of information on rotavirus disease burden in sub-Saharan Africa. METHODS: We conducted prospective, hospital-based surveillance for rotavirus diarrhea among children <5 years of age at the tertiary care Dr. George Mukhari Hospital (DGM) and at the Brits district Hospital (BH) in the Gauteng and North West Provinces in South Africa; we estimated that up to 80% of children <5 years of age in their catchment areas who are hospitalized for diarrhea are admitted to one of these hospitals. RESULTS: At DGM, 2553 children <5 years of age were admitted for diarrhea from January 2003 through December 2005, and 852 children <5 years of age were treated for diarrhea at BH during 2004-2005. We examined stool specimens from 450 children (53%) at BH and from 1870 children (73%) admitted to DGM. An estimated 22.8% (95% confidence interval [CI], 21.2%-24.5%) of the children hospitalized with diarrhea at DGM were rotavirus positive, and the corresponding figure at BH was 18.2% (95% CI, 14.9%-22.1%). Among children <5 years of age admitted to DGM for any reason, an estimated 5.5% (95% CI, 5.1%-6.0%) had rotavirus diarrhea. Our incidence estimates suggest that 1 in 43-62 children in the area is likely to be hospitalized with rotavirus diarrhea by 2 years of age. CONCLUSIONS: Prevention of serious rotavirus illness by vaccination will substantially reduce not only the disease burden among young children but also the case load in South African health care facilities.

Determination of the G and P types of previously nontypeable rotavirus strains from the African Rotavirus Network, 1996-2004: Identification of unusual G types.

A total of 215 nontypeable rotavirus samples collected from children <5 years of age by members of the African Rotavirus Network were characterized using reverse-transcription polymerase chain reaction analysis and sequencing. The most predominant strain identified was P[8]G1 (46.9%). Genotypes P[8]G10, P[8]G8, P[6]G8, and P[7]G5 were also detected at frequencies varying from 0.5% to 2.3%. This study suggests that reassortment of unusual G types into a background of globally common genotype P[8] strains may be a major mechanism of generating rotavirus diversity. Nucleotide substitutions at the P[8], P[6], and G1 primer binding sites accounted for the failure to type these strains initially. Hence, these findings highlight the need for regular evaluation of rotavirus genotyping methods.

Whole Genome Analysis of African G12P[6] and G12P[8] Rotaviruses Provides Evidence of Porcine-Human Reassortment at NSP2, NSP3, and NSP4.

Group A rotaviruses (RVA) represent the most common cause of pediatric gastroenteritis in children <5 years, worldwide. There has been an increase in global detection and reported cases of acute gastroenteritis caused by RVA genotype G12 strains, particularly in Africa. This study sought to characterize the genomic relationship between African G12 strains and determine the possible origin of these strains. Whole genome sequencing of 34 RVA G12P[6] and G12P[8] strains detected from the continent including southern (South Africa, Zambia, Zimbabwe), eastern (Ethiopia, Uganda), central (Cameroon), and western (Togo) African regions, were sequenced using the Ion Torrent PGM method. The majority of the strains possessed a Wa-like backbone with consensus genotype constellation of G12-P[6]/P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1, while a single strain from Ethiopia displayed a DS-1-like genetic constellation of G12-P[6]-I2-R2-C2-M2-A2-N2-T2-E2-H2. In addition, three Ethiopian and one South African strains exhibited a genotype 2 reassortment of the NSP3 gene, with genetic constellation of G12-P[8]-I1-R1-C1-M1-A1-N1-T2-E1-H1. Overall, 10 gene segments (VP1-VP4, VP6, and NSP1-NSP5) of African G12 strains were determined to be genetically related to cognate gene sequences from globally circulating human Wa-like G12, G9, and G1 strains with nucleotide (amino acid) identities in the range of 94.1-99.9% (96.5-100%), 88.5-98.5% (93-99.1%), and 89.8-99.0% (88.7-100%), respectively. Phylogenetic analysis showed that the Ethiopian G12P[6] possessing a DS-1-like backbone consistently clustered with G2P[4] strains from Senegal and G3P[6] from Ethiopia with the VP1, VP2, VP6, and NSP1-NSP4 genes. Notably, the NSP2, NSP3, and NSP4 of most of the study strains exhibited the closest relationship with porcine strains suggesting the occurrence of reassortment between human and porcine strains. Our results add to the understanding of potential roles that interspecies transmission play in generating human rotavirus diversity through reassortment events and provide insights into the evolutionary dynamics of G12 strains spreading across selected sub-Saharan Africa regions.

Outbreak of diarrhoea in piglets caused by novel rotavirus genotype G4P[49] in north-western district of Bangladesh, February 2014.

Group A rotavirus (RVA) associated diarrhoea in piglets represents one of the major causes of morbidity and mortality in pig farms worldwide. A diarrhoea outbreak occurred among nomadic piglets in north-western district of Bangladesh in February 2014. Outbreak investigation was performed to identify the cause, epidemiologic and clinical features of the outbreak. Rectal swabs and clinical information were collected from diarrhoeic piglets (n = 36). Rectal swabs were tested for RVA RNA by real-time reverse transcription polymerase chain reaction (rRT-PCR) using NSP3-specific primers. The G (VP7) and P (VP4) genes were typed by conventional RT-PCR and sanger sequencing and full genome sequences were determined using next-generation sequencing. We found the attack rate was 61% (50/82) among piglets in the nomadic pig herd, and the case fatality rate was 20% (10/50) among piglets with diarrhoea. All study piglets cases had watery diarrhoea, lack of appetite or reluctance to move. A novel RVA strain with a new P[49] genotype combined with G4 was identified among all piglets with diarrhoea. The genome constellation of the novel RVA strains was determined to be G4-P[49]-I1-R1-C1-M1-A8-N1-T7-E1-H1. Genetic analysis shows that the novel G4P[49] strain is similar to Indian and Chinese porcine or porcine-like G4 human strains and is genetically distant from Bangladeshi human G4 strains. Identification of this novel RVA strain warrants further exploration for disease severity and zoonotic potential.

Full genome characterization of human Rotavirus A strains isolated in Cameroon, 2010-2011: diverse combinations of the G and P genes and lack of reassortment of the backbone genes.

Over the past few years whole genome sequencing of rotaviruses has become a routine laboratory method in many strain surveillance studies. To study the molecular evolutionary pattern of representative Cameroonian Rotavirus A (RVA) strains, the semiconductor sequencing approach was used following random amplification of genomic RNA. In total, 31 RVA strains collected during 2010-2011 in three Cameroonian study sites located 120 to 1240 km from each other were sequenced and analyzed. Sequence analysis of the randomly selected representative strains showed that 18 RVAs were Wa-like, expressing G1P[6], G12P[6], or G12P[8] neutralization antigens on the genotype 1 genomic constellation (I1-R1-C1-M1-A1-N1-T1-E1-H1), whereas 13 other strains were DS-1-like, expressing G2P[4], G2P[6], G3P[6], and G6P[6] on the genotype 2 genomic constellation (I2-R2-C2-M2-A2-N2-T2-E2-H2). No inter-genogroup reassortment in the backbone genes was observed. Phylogenetic analysis of the Cameroonian G6P[6] strains indicated the separation of the strains identified in the Far North region (Maroua) and the Northwest region (Bamenda and Esu) into two branches that is consistent with multiple introductions of G6P[6] strains into this country. The present whole genome based molecular characterization study indicates that the emerging G6P[6] strain is fully heterotypic to Rotarix, the vaccine introduced during 2014 in childhood immunization program in Cameroon. Continuous strain monitoring is therefore needed in this area and elsewhere to see if G6s, besides genotype G1 to G4, G8, G9 and G12, may become a new, regionally important genotype in the post vaccine licensure era in Africa.

Shared G12 VP7 gene among human and bovine rotaviruses detected in Cameroonian villages.

Group A rotaviruses (RVA) are an important enteric pathogen in humans and livestock animals. Transmission of animal RVA strains to humans has been documented on several occasions. A reverse route of transmission of RVA under natural circumstances is anticipated, although evidence is scarce. However, experimental studies indicated that animals can be infected with human RVAs. By screening the stool samples collected from 157 cattle during 2011 in two Cameroonian villages, four samples (2.5%) were found positive for RVA. Upon sequence analysis of a 410 bp fragment of the VP7 gene, the RVA strains shared up to 100% nt identity to each other and to G12 RVAs identified in human patients living in the same geographic regions. This finding provides evidence for a human-to-animal transmission of an epidemic human rotavirus strain.

Detection of novel porcine bocaviruses in fecal samples of asymptomatic pigs in Cameroon.

Improvements and widespread use of nucleic acid amplification and sequencing methods have led to the recognition of new virus diversity in various domestic animals, including pigs. In this study we utilized either virus species specific or broadly reactive PCR assays to describe the occurrence and genetic diversity of selected DNA viruses belonging to families Adenoviridae, Circoviridae, Anelloviridae and Parvoviridae in Cameroonian pigs. Fecal specimens were collected during spring of 2011. No adenoviruses, circoviruses and anelloviruses were detected, however, high prevalence and remarkable genetic diversity within the identified parvoviruses and, particularly, within bocaviruses was observed. PPV4 was the most prevalent virus (20%), followed by PBoV3 (18%), PBoV4 (18%), PBoV5 plus 6V/7V (16%), and PBoV1 plus PBoV2 (6%). The frequency of mixed infections with various combinations of these virus species reached 20%. Genetic analysis of the identified viruses showed that the capsid gene of PBoV1 and PBoV2 strains shared up to 91% and 94%nt sequence similarities to reference PBoV1 and PBoV2 strains, respectively. The identified PBoV3 and PBoV4 strains shared ≤ 95% and ≤ 98%nt identities with reference PBoV3 and PBoV4 strains, respectively, along the NS gene, whereas the PBoV5 strains shared 86%nt identities with Hungarian and 87%nt identities with Chinese PBoV5 strains along the capsid gene. In addition, a single PBoV5-like strain shared ≤ 71%nt sequence identity with other PBoV5 strains. This is the first study to report evidence of the circulation of bocaviruses in Africa and contributes to our understanding of the impact of globalization on the dispersal of new and emerging viruses.

Epidemiology of rotavirus diarrhea in children under 5 years in Northern Cameroon.

BACKGROUND: Rotavirus still remains the major cause of diarrhea in children below 5 years. No data on rotavirus epidemiology is available in the Northern regions of Cameroon. We aimed to determine the prevalence of group A rotavirus (RVA) in children below 5 years with diarrhea in two regions of Northern Cameroon (North West and Far North Regions) so as to improve our knowledge on the burden of rotavirus disease for imminent introduction of a rotavirus vaccine. METHODS: Stool samples were collected during 2010 and 2011 from 390 children below 5 years presenting with diarrhea in four hospitals in Northern Cameroon and were screened for rotavirus group A by reverse transcription-polymerase chain reaction. RESULTS: This study revealed that 42.8% of the children below 5 years had group A rotavirus infection, 46.5% in the Far North region while the North West had a prevalence of 33.9%. Of the 252 hospitalized and the 138 outpatient children, 124(49.2%) and 43(31.2%) (P=0.00085), respectively, were positive for group A rotavirus. Children below 24 months were most affected (44.7%), while the age group 49-60 months had the lowest prevalence (25%). The RVA prevalence was 44.6% in the urban and 28.9% in the rural settings of our study. It was observed that the proportion of children with diarrhea who had rotavirus accompanied with fever and vomiting in the outpatient group and inpatient group were 13.0% and 28.6% respectively, P=0.03. CONCLUSION: This study showed high incidence of rotavirus infection especially among hospitalized children in Northern Cameroon, suggesting that rotavirus is a major cause of childhood morbidity and mortality in this area.