Virology
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Item A possible risk of environmental exposure to HEV in Ibadan, Oyo State, Nigeria(Taylor & Francis, 2020-08-13) Olayinka, A.; Ifeorah, I. M.; Omotosho, O.; Faleye, T. O. C.; Odukaye, O.; Bolaji, O.; Ibitoye, I.; Ope-Ewe, O.; Adewumi, M. O.; Adeniji, J. A."Hepatitis E virus (HEV) infection is both a major public health concern and emerging global health concern, with a documented incidence of 20 million, 3.4 million clinical cases, 70,000 deaths, and 3,000 stillbirths. The aetiologic agent, HEV is a primarily enterally transmitted hepatotropic virus. Fecal samples were collected from three selected pig farms across Ibadan, South-west Nigeria. Randomly picked samples were pooled per unit pen and fecal suspensions prepared were subjected to HEV Antigen (Ag) enzyme-linked immunosorbent assay. Molecular probing was done by Reverse Transcription and nested polymerase reaction (RT-nPCR) and deep sequencing. Sequencing was done paired-end for 300 cycles using the HiSeq system. Overall farm prevalence of 66.7% (2/3) and prevalence at individual level of 13.2% (9/68) were recorded. All nine samples positive for the ELISA screen were negative when subjected to RT-nPCR assays. Further, on deep sequencing, no HEV genomic fragment was found in the sample using de-novo assembly. Findings suggest possibly inapparent HEV in the pigs studied or a yet to be identified protein with HEV-Ag cross-reactivity ability on ELISA, thus constituting a possible risk of exposure to HEV infection in the population. Consequently, we recommend prompt intervention to unravel the mystery and break the chain of transmission.Item Extending the utility of the WHO recommended assay for direct detection of enteroviruses from clinical specimen for resolving poliovirus co‑infection(BMC Research Notes, 2018) Faleye, T. O. C; Adewumi, M. O.; Ozegbe, N. P.; Ogunsakin, O. E.; Ariyo, G.; Adeshina, F. W.; Ogunga, O. S.; Oluwadare, S. D.; Adeniji, J. A.Objectives: In a polio-free world there might be reduced funding for poliovirus surveillance. There is therefore the need to ensure that enterovirologist globally, especially those outside the global polio laboratory network, can participate in poliovirus surveillance without neglecting their enterovirus type of interest. To accomplish this, assays are needed that allow such active participation. Results: In this study we describes a sensitive and specific utility extension of the recently recommended WHO RT-snPCR assay that enables independent detection of the three poliovirus types especially in cases of co-infection. More importantly, it piggy-backs on the first round PCR product of the WHO recommended assay and consequently ensures that enterovirologists interested in nonpolio enteroviruses can continue their investigations, and contribute significantly and specifically to poliovirus surveillance, by using the excess of their first round PCR product.Item The fallacy of identification by neutralization in the light of non-cytopathic effect producing enterovirus strains(SCIENCEDOMAIN International, 2017) Adewumi, M. O.; Faleye, T. O. C.; Adeniji, J. A.We describe the characterization of an enterovirus isolate recovered from untreated raw sewage in Ibadan, southwest Nigeria in 2010. The isolate was neutralized by specific antisera and consequently identified as Echovirus 7 (E7). Subsequent molecular characterization showed the isolate to be a mixture of E7 and Coxsackievirus A24 (CV-A24) thereby suggesting the CV-A24 strain to be non-cytopathic effect producing. It is therefore crucial that identities of enterovirus strains determined by neutralization assay be verified by molecular characterization to ensure they do not have any non-cytopathic effect producing strain(s) lurking unnoticed.Item Isolation and Identification of Enteroviruses from Sewage and Sewage-Contaminated Water Samples from Ibadan, Nigeria, 2012-2013(SciTechnol, 2017) Adeniji, J. A.; Adewale, A. O.; Faleye, T. O. C.; Adewumi, M. O.In 2010, we described sewage contaminated water (SCW) bodies that consistently yielded enteroviruses (EVs) in enterovirus surveillance (ES) sites in Lagos, Nigeria. By 2012, we demonstrated the presence and circulation of Wild Poliovirus 3 (WPV3) in these ES sites. Here we describe ES sites that consistently yield EVs in Ibadan metropolis southwest Nigeria. Twenty-five ES samples were collected by grab method from nine sites between October, 2012 and March, 2013. Samples were concentrated and four (RD, HEp2C, MCF-7 and L20B) different cell lines used for virus isolation from the concentrates. Isolates were subjected to RNA extraction, cDNA synthesis, PanEnterovirus 5l-UTR and VP1 assays. Unidentifiable isolates were further subjected to species-specific RTPCR assays. Amplicons were sequenced, isolates identified and subjected to phylogenetic analysis. Twenty-five isolates were recovered from 8 (32%) of the samples collected. Twenty-three of the isolates were identified as EVs by the PanEntero5l-UTR assay. Thirteen (57%) of the 23 EVs were positive for the VP1 assay, and identified as Coxsackievirus B3 (CVB3) (1 isolate), CVB6 (1 isolate), E6 (2 isolates), E7 (5 isolates), E11 (1 isolate), E12 (1 isolate) and E13 (2 isolates). None and 2 (25%) of the remaining isolates were positive for the EV-B and EV-C assays, respectively. The 2 EV-C positive enteroviruses were isolated on MCF-7. This study describes three very productive ES sites, and documents the presence of CVB3, CVB6, E6, E7, E11, E12 and, E13 in Ibadan, Nigeria. Including other cell lines in EV isolation protocols can broaden the diversity of EV types recoverable.Item Development of Echovirus 29 Cytopathology in RD Cell Line Might not Happen within 10 Days Post Inoculation(SciTechnol, 2017) Adewumi, M. O.; Faleye, T. O. C.; Ayinde, O. T.; Ibok, U. I.; Adeniji, J. A.Classically in enterovirology, isolation in RD cell culture required inoculation and incubation for 14 days before any sample is declared negative for enteroviruses. To reduce the turn-around time for confirming suspected cases of poliomyelitis, in 2010, the required incubation period, post inoculation was reduced to 10days. In this study we set out to determine why an Echovirus 29 (E29) strain we isolated in 2016 showed no cytopathology on RDcell culture despite the fact that other members of the clade to which it belongs do so. We found that the problem was not the 2016 E29 strain but the new 10 days incubation algorithm. To be precise, the 2016 E29 strain did not show cytopathology in RD cell line within the recommended 10 days but replicated with evident cytopathology when allowed to stay in culture for 13 to 14 days. This shows that some samples declared negative for enteroviruses by the 10 days incubation algorithm might be false negatives that will eventually develop cytopathology if allowed to remain in incubation for 13 to 14 days. It is therefore encouraged that those particularly interested in Non-Polio Enteroviruses endeavour to maintain at least 14 days incubation in cell culture in a bid to accommodate the likes of E29 that might need 13-14 days to develop CPE especially when present at low titre.Item Profiles of Hepatitis B Virus Serological Markers among Asymptomatic Population in Anambra State, Southeastern Nigeria(SciTechnol, 2017) Bakarey, A. S.; Ifeorah, I. M.; Adewumi, M. O.; Faleye, T. O. C.; Akere, A.; Omoruyi, C. E.; Ogunwale, A. O.; Uttah, C. C.; Oketade, M. A.; Adeniji, J. A.Hepatitis B Virus (HBV) infection is apparent in endemic countries affecting millions of people. Further, the asymptomatic nature of the pathogen is a major public health concern. This study was designed to assess the burden of HBV by exploring the serologic markers of infection among consenting asymptomatic community dwellers in two cities in southeastern Nigeria. A total of 405 blood specimens were tested for HBsAg, anti-HBs, HBeAg, anti-HBe, total anti-HBc and anti-HBc-IgM using ELISA technique. Overall, 14(3.5%) of the participants had detectable HBsAg out of which 1 (7.1%) had HBeAg and 13, anti-HBe. Two of the HBsAg positives (14.3%) had detectable anti-HBc-IgM. A total of 144 (35.5%) had detectable anti-HBc, even as 65 (57.0%) of them had the marker as the only serologic evidence of HBV exposure. Thirty-seven (9.1%) participants had anti-HBs only although all of them were born before the start of the childhood HBV vaccination. Altogether, 224 (57.3%) had no detectable serological markers of HBV infection or immunity and were obviously at risk ofHBV infection. This study described various patterns of HBV serologic markers of infection in the study population and probable risk of viru spread. Our results support the need for urgent intervention and implementation of measures to control the spread of HBV infection in Nigeria.Item Patterns of serologic markers of hepatitis B virus infection and the risk of transmission among pregnant women in southwestern Nigeria(Taylor & Francis, 2017) Ifeorah, I. M.; Bakarey, A. S.; Adewumi, M. O.; Faleye, T. O. C.; Akere, A.; Omoruyi, C. E.; Ogunwale, A. O.; Uttah, C. C.; Oketade, M. A.; Adeniji, J. A.Hepatitis B virus (HBV) infection is a major health concern in developing countries that has a high morbidity and mortality rate. Vertical transmission of HBV from mother to child has been identified as a major factor leading to chronicity with attendant liver conditions, especially in poor socioeconomic settings. This study aims to evaluate the prevalence of serological HBV markers among pregnant women in Ibadan southwestern Nigeria and to determine the implications for perinatal HBV transmission. This study revealed the presence of varied HBV serological patterns of infection or immunity among pregnant women in Ibadan, Nigeria, and thus the risk of mother to child transmission.Item Non-polio enteroviruses in faeces of children diagnosed with acute flaccid paralysis in Nigeria(Springer Nature, 2017) Faleye, T. O. C.; Adewumi, M. O.; Japhet, M. O.; David, O. M.; Oluyege, A. O.; Adeniji, J. A.; Famurewa, O.The need to investigate the contribution of non-polio enteroviruses to acute flaccid paralysis (AFP) cannot be over emphasized as we move towards a poliovirus free world. Hence, we aim to identify non-polio enteroviruses recovered from the faeces of children diagnosed with AFP in Nigeria.Ninety-six isolates, (95 unidentified and one previously confirmed Sabin poliovirus 3) recovered on RDcell culture from the stool of children <15 years old diagnosed with AFP in 2014 were analyzed. All isolates were subjected to RNA extraction, cDNA synthesis and three different PCR reactions (one panenterovirus 5′-UTR and two different VP1 amplification assays). VP1 amplicons were then sequenced and isolates identified. Results: 92.71% (89/96) of the isolates were detected by at least one of the three assays as an enterovirus. Precisely, 79.17% (76/96), 6.25% (6/96), 7.30% (7/96) and 7.30% (7/96) of the isolates were positive for both, positive and negative, negative and positive, as well as negative for both the 5′-UTR and VP1 assays, respectively. In this study, sixty-nine (69) of the 83 VP1 amplicons sequenced were identified as 27 different enterovirus types. The most commonly detected were CV-B3 (10 isolates) and EV-B75 (5 isolates). Specifically, one, twenty-four and two of the enterovirus types identified in this study belong to EV-A, EV-B and EV-C respectively. This study reports the circulating strains of 27 non-polio enterovirus types in Nigerian children with AFP in 2014 and Nigerian strains of CV-B2, CV-B4, E17, EV-B80, EV-B73, EV-B97, EV-B93, EV-C99 and EV-A120 were reported for the first time. Furthermore, it shows that being positive for the 5′-UTR assay should not be the basis for subjecting isolates to the VP1 assays.Item High Prevalence of Anti-HCV Antibodies Among Pregnant Women in Southwestern Nigeria(IISTE, 2017) Japhet, M. O.; Donbraye, E.; Adesina, O. A.; Adewumi, M. O.Hepatitis C virus (HCV) is the most common cause of cirrhosis, hepatocellular carcinoma and liver transplantation. While universal screening for other blood-borne viruses (BBVs) such as HIV and HBV among pregnant women is recommended in Nigeria, no such recommendation exist for HCV in the country. Despite recently developed direct-acting antiviral agents (DAAs) to cure HCV at high rates and at very high cost, the absence of an HCV vaccine or approved therapy during pregnancy makes prevention of vertical transmission impossible at the moment. Using a commercially available enzyme linked immunoassay technique, prevalence of antibodies to hepatitis C virus (anti-HCV) was determined among pregnant women attending antenatal clinics in Southwestern Nigeria. Of the 273 serum samples obtained from the pregnant women 9.5% was positive for anti-HCV antibody. There were differences in anti-HCV prevalence by age and locality. Results of the study confirm endemicity of HCV among pregnant women in the country, consequently, we advocate free screening, among other essential measures for HCV intervention in Nigeria.Item Hepatitis B Virus Serological Markers in a Rural Community in Southeastern Nigeria(SCIENCEDOMAIN International, 2017) Bakarey, A. S.; Ifeorah, I. M.; Faleye, T. O. C.; Adewumi, M. O.; Akere, A.; Omoruyi, C. E.; Ogunwale, A. O.; Olaleye, V. F.; Awokunle, R. F.; Sekoni, D.E.Due to the current blood safety algorithm in Nigeria which excludes only Hepatitis B surface Antigenaemia (HBsAg) positive individuals from blood donation, this study was therefore designed to investigate HBV markers of infection in a rural population in southeastern Nigeria. It is a cross sectional community-based study. This study was carried out in Awuda village in Nnobi town of southeastern Nigeria in August 2013. A total of 92 consenting participants were enrolled for the study. The participants were screened for HBsAg, HBeAg, Anti-HBc IgM, Anti-HBc, Anti-HBe and Anti-HBs using ELISA technique and classified into different serological profiles indicative of infection stages. Respondents’ mean age was 26.3(SD +11.5) years and 58 of them were females while 34 were males. An overall prevalence for HBsAg was 1.1% (1/92). The same HBsAg positive individual also had detectable anti-HBe and anti- HBc IgM. Analysis of the results showed 3 (3.3%) of the study participants were positive for both Anti-HBe and Anti-HBc. Also, 12 (13.0%) participants were positive for only Anti-HBc and Anti-HBs antibody. Another 9 (9.8%) participants were positive for only Anti-HBs while 51 (55.0%) had no serological marker for previous exposure to either HBV or HBV vaccine. Altogether, 31(33.7%), 1(1.1%) and 21(22.8%) participants were positive for HBc, HbcIgM and HBs antibodies respectively. This study has demonstrated that at least 1 out of every 3 people in the studied community might have serological evidence of present or past HBV infection. The current dependence of blood safety algorithms which excludes only HBsAg positive individuals is not enough to guarantee safety of blood and/or blood products. More studies are needed to further investigate the theoretical basis of the algorithm.
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