Virology
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Item Circulation of hepatitis B virus genotype-E among outpatients in tertiary hospitals in the Niger-Delta region of Nigeria(African Journals Online (AJOL), 2022) Umego, C. F.; Mboto, C. I.; Asitok, A. D.; Osaji, L. C.; George, U. E.; Edet, U. O.; Mbim, E. N.; Faleye, T. O. C.; Adewumi, O. M.; Adeniji, J. A.Introduction: Hepatitis B virus (HBV) infection continues to be a significant public health challenge globally, with higher disease burden in developing countries. HBV genotypes are associated with different geographical regions and clinical outcomes. Limited information exists on epidemiology of HBV in the Niger-Delta region (South-South) of Nigeria. Consequently, this study was designed to characterise hepatitis B virus infection among outpatients in selected tertiary hospitals in the region. Methodology: Between June and August 2017, consenting nine hundred asymptomatic out-patients were enrolled and initially screened for HBV infection using one step Hepatitis B surface antigen (HBsAg) strip and subsequently re-tested using HBsAg and Hepatitis B core total antibody (anti-HBc) specific Enzyme-Linked Immunosorbent Assay (ELISA). Blood serum with detectable HBsAg were subsequently subjected to DNA extraction, S-gene amplification using a nested polymerase chain reaction (PCR) protocol, gel electrophoresis, sequencing and phylogenetic analysis. Results: Seroprevalence of HBsAg was 4.6% (95% CI 2.5-7.1) and anti-HBc was 10.1% (95% confidence interval (CI) 6.1-15.3). Of the 41 HBsAg positive samples subjected to DNA extraction and HBV S-gene specific PCR, only 6 (14.6%) yielded the expected ~408bp band. Phylogenetic analysis based on HBV pre-S/S sequences identified all six typable samples as genotype E, subtype ayw4 of the West African clade. Conclusion: Results of the study confirm the presence and circulation of HBV genotype-E in the Niger-Delta region of Nigeria, thus corroborating the inclusion of the country in the Genotype E crescent. The authors advocate value-added HBV intervention in the region and the country at large.Item Prevalence of hepatitis b virus (HBV) basal core promoter/precore region molecular variants among HIV/HBV co-infected and HBV mono-infected patients in Ile-Ife, Nigeria(Medwin Publishers, 2021) Osasona, O. G.; Boudarene, L.; Fasoro, O. A.; George, U. E.; Oguzie, J.; Ariyo, O. E.; Olumade, T. J.; Adeniyi, A.; Adewumi, O. M.; Adeniji, J. AIntroduction: Evolution of phenotypic diversity among viruses occurs as an escape mechanism against host immune pressure or drug selective pressure. Among HIV/HBV co-infected individuals, various HBV basal core promoter (BCP)/precore (PC) region molecular mutants had been reported with associated phenotypic defect in HBeAg production. The emergence of HBeAg negative variants of HBV in HIV co-infected individuals have profound implication on the diagnosis, management and prognosis of this subset of individuals. This includes delayed clearance of HBV, early development of adverse hepatic events such as liver cirrhosis and hepatocellular carcinoma. Currently, little is known about HBV BCP/PC region genomic heterogeneity in HIV/HBV co-infected patients in Nigeria. Therefore, this study was focused on investigating evidence of precore/core region genomic variability among HIV/HBV co-infected patients in Nigeria. Materials and Methods: A total of 40 patients (20 HIV/HBV co-infected and 20 HBV mono-infected samples) were enrolled into the study and subsequently tested for HBsAg, HBeAg and HBeAb using specific Enzyme-Linked Immunosorbent Assay (ELISA). The BCP/PC genome regions (nucleotides 1653-1959) were amplified using a nested PCR assay and then subjected to BCP/PC mutational analysis in genome sites affecting HBeAg expression especially at the BCP transcriptional and PC Translational stop codon sites. Results: Overall, 5 (83.3%) of the six exploitable sequences after analysis showed various BCP/PC mutations. Only 1(16.6%) sequence from an HIV/HBV co-infected patient had the BCP transcriptional (double mutation; A1762T/G1764A) mutant. Analysis of the PC translational stop codon showed 4 (66.6%) having the G1896A mutants while 33.3% (2) had G1899A mutants. Conclusion: This study has broadened the available evidence of BCP/PC region molecular mutants among HIV/HBV coinfected patients in Nigeria and assessed the difference of mutation prevalence in comparison with HBV mono-infected cohort.Item Isolation and Genomic Characterization of Echovirus 11 from faeces of a Non-Human Primate in Nigeria(Springer Nature, 2021-05-16) Faleye, T. O. C.; George, U. E.; Klapsa, D.; Majumdar, M.; Oragwa, A. O.; Adewumi, O. M.; Martin, J.; Adeniji, J. A.We recently investigated the presence of enteroviruses (EVs) in non-human primates (NHPs) in Northern Nigeria and documented the presence of EV-A76 of South-East Asian ancestry in an NHP. In this study, we go further to ask if we could also find EVs in NHPs indigenous to the forested South-south Nigeria. Fresh faecal samples were collected from the floor of 10 cages housing NHPs in Cross River Nigeria, resuspended in PBS and subjected to RNA extraction, cDNA synthesis, PanEnt 50-UTR and PanEnt VP1 PCR assays. None of the samples was positive for the PanEnt VP1 assay, but one sample was positive for PanEnt 50- UTR PCR. This sample was subsequently inoculated into RD cell line, produced CPE and the isolate analysed by PCR assays, next-generation whole genome sequencing and passage in four different cell lines showing replication in two of them. Analysis of the complete genome of the isolate identified it as an Echovirus 11 (E11) and revealed a recombinant genomic structure. Phylogenetic analysis showed that the E11 NHP strain was related to human clinical isolates suggesting a zoonotic behaviour. We describe the first isolation and complete genome characterization of an E11 obtained from an NHP in Nigeria having zoonotic potential.Item Fecal Antibiotic Resistome of Pigs from a Small-Scale Piggery in Ibadan, South-West Nigeria(ResearchersLinks (Nexus Academic Publishers, NAP), 2021-05-28) Olayinka, O. A.; Faleye, T. O. C.; Omotosho, O. O.; Odukaye, O. A.; Oluremi, B.; Ibitoye, I. H.; Ope-Ewe, O. O.; George, U. E.; Arowolo, O. A; Ifeorah, I. M.; Omoruyi, E. C.; Donbraye, E; Adeniji, O. P.; Adewumi, O. M.; Adeniji, J. A.This study was designed to sample the fecal Resistome of Pigs from a small-scale Piggery in Ibadan, South-West Nigeria. Three fecal pellets were randomly picked from the floor of unit pens containing a minimum of three pigs per unit in selected piggery in Ibadan, Nigeria. The samples were pooled and resuspended in phosphate buffered saline. The suspension was then subjected to nucleic acid extraction, Cdna synthesis and Illumina sequencing. Antibiotic Resistance Genes (ARGs) in the raw reads were determined and assembled using the Kmer Resistance tool v2.2. From the 2,974,257 reads generated, 21 ARGs with statistically significant reads were identified. Almost all targeted broad-spectrum antibiotics with over 50% targeting Tetracyclines. Five (ant(6)-Ia_3, tet(40)_1, tet(Q)_1, tet(W)_5 and tet(O/W)_4) of the ARGs were predicted to be plasmid-borne. Our findings show that the Swine industry in the region might be both a mixing pot and reservoir of ARGs. It is therefore crucial that effort is made to educate the stakeholders on the importance of good antibiotics stewardship.Item Faecal antibiotic resistome of Nigerian chimpanzees from a wildlife sanctuary in cross-river state, Nigeria(Nexus Academic Publishers (NAP), 2021) George, U. E.; Arowolo, O. A.; Olayinka, O. A.; Ifeorah, I. M.; Faleye, T. O. C.; Oluremi, B.; Oragwa, A.O.; Omoruyi, E. C.; Udoh, E. E.; Osasona, O. G.; Donbraye, E.; Adeniji, O. P.; Adewumi, O. M.; Adeniji, J. A."Out of prescription, indiscriminate use, misuse and abuse of antibiotic administration and/or usage in both humans and farm animals have led to a consistent interference and contamination of biomes and ecobiomes. These resultantly give platforms for positive selection of resistant pathogens and high levels of Antibiotic Resistance Genes (ARGs). We examined Nigerian Chimpanzees in Boki Afi Mountain Wildlife Sanctuary, Cross-River State, Nigeria, to detect ARGs. Faecal samples from 15 Chimpanzees in pristine enclosures of Non-Human Primates in the Wildlife Sanctuary were analyzed. All faecal samples were pooled, then resuspended in phosphate-buffered saline. Subsequently, nucleic acid was extracted from the suspension and Illumina sequencing performed. ARGs in the raw reads were determined and assembled using the KmerResistance tool v2.2. From the 2,763,954 reads generated, 14 ARGs with statistically significant reads were identified. Precisely, 90.5% (12/14) of the ARGs detected target drugs that inhibit translation, of which 66.6% (8/12) were tetracycline resistance (TC-r) genes, while remaining 9.5% (2/14) inhibit cell wall synthesis (cfxA3_1 and cfxA6_1). Eight (aph(3’)-III_1, cfxA3_1, cfxA6_1, erm(B)_10, tet(Q)_1, tet(Q)_2, tet(Q)_4, tet(W)_5) of the ARGs detected were predicted to be plasmid-borne. We report using a cultivation-independent approach the presence of ARGs in Nigerian Chimpanzees. Findings suggest Nigerian Chimpanzees may constitute a hitherto overlooked source of antibiotic resistance in the environment. These ARGs may have been exchanged with handlers and rural dwellers around the Sanctuary. Surveillance of sympatric human faecal and environmental microbiota and their resistomes at the Wildlife Sanctuary are merited to inform public health interventions and decrease ARGs dissemination."Item Near-complete genome sequence of an enterovirus species fisolate recovered from sewage in Nigeria(American Society for Microbiology (ASM), 2021) Faleye, T. O. C.; Ifeorah, M. I.; Olayinka, O. A.; Oluremi, B.; George, U. E.; Arowolo, O. A.; Omoruyi, E. C.; Donbraye, E.; Oyero, A. O.; Adewumi, O. M.; Adeniji, J.A.Here, we describe the near-complete genome of an enterovirus F (EV-F) isolate from Nigeria. The obtained sequence was 7,378 nucleotides (nt) long and encodes 2 open reading frames (ORFs), an upstream ORF (uORF; 56 amino acids [aa])and a polyprotein ORF (ppORF; 2,167 aa). Both ORFs overlap but are in different reading frames, with the uORF in a _1 reading frame relative to the ppORF.Item Draft genome sequence of a bovine enterovirus isolate recovered from sewage in Nigeria(American Society for Microbiology (ASM), 2018) Faleye, T. O. C; Adewumi, O. M.; Olayinka, O. A.; Donbraye, E; Oluremi, B.; George, U. E.; Arowolo, O. A.; Omoruyi, E. C.; Ifeorah, M. I.; Oyero, A . O.; Adeniji, J. A.We describe the draft genome of a bovine enterovirus (EV) isolate recovered from sewage in Nigeria. This isolate replicates on both RD and L20B cell lines but is negative for all EV screens in use by the Global Poliovirus Eradication Initiative (GPEI). It contains 7,368 nucleotides (nt) with 50.2% G C content and an open reading frame (ORF) with 6,525 nt (2,174 amino acids).Item Recovery of nonpolio enteroviruses from l20b cell line with non-reproducible cytopathic effect(BioMed Central, 2018) Adeniji, J. A.; Ibok, U. I.; Ayinde,O. T.; Oragwa, A. O.; George, U. E.; Faleye. T. O. C.; Adewumi, M. O.Background and Aim of the Study: Samples showing cytopathic effect (CPE) on initial inoculation into L20B cell line but with no observed or reproducible CPE on passage in L20B or RD are considered negative for both poliovirus and nonpolio enteroviruses (NPEVs). The phenomenon is termed ‘non-reproducible CPE’. Its occurrence is usually ascribed to the likely presence of reoviruses, adenoviruses and other non-enteroviruses. This study aimed to investigate the likelihood that NPEVs are also present in cases with non-reproducible CPE. Place and Duration of Study: This study was carried out in the Department of Virology, College of Medicine, University of Ibadan using twenty-six (26) cell culture suspensions collected from the WHO National Polio Laboratory, Department of Virology, College of Medicine, University of Ibadan. The suspensions emanated from 13 L20B cell culture tubes that showed cytopathology within 5 days of inoculation with faecal suspension from AFP cases. However, on passage into one each of RD and L20B cell lines, the CPE was not reproducible. The study lasted for three (3) months from samples collection to report writing. Methodology: All samples were subjected to RNA extraction, cDNA synthesis, the WHO recommended VP1 RT-seminested PCR assay, species resolution PCR assay, sequencing andphylogenetic analysis. Results: Six (6) samples were positive for the VP1 RT-seminested PCR assay. Only four of which were positive by the species resolution PCR assay. The four amplicons were sequenced, however, only three (3) were successfully identified as Coxsackievirus A20 (2 isolates) and Echovirus 29 (1isolate). Conclusion: The results of this study unambiguously showed the presence of NPEVs (particularly CVA20 and E29) in cell culture supernatants of samples with CPE on initial inoculation into L20B cell line but with no observed or reproducible CPE on passage in RD cell line. Therefore, like reoviruses, adenoviruses and other non-enteroviruses, NPEVs can also be recovered in cases with non-reproducible CPE.Item Preponderance of enterovirus C in RD-L20B-cell-culture-negative stool samples from children diagnosed with acute flaccid paralysis in Nigeria(Springer Nature, 2017) Adeniji, J. A.; Oragwa, A. O.; George, U. E.; Ibok, U. I.; Faleye, T. O. C.; Adewumi, M. ORecently, a reverse transcriptase semi-nested polymerase chain reaction (RT-snPCR) assay was recommended by the WHO for direct detection of enteroviruses in clinical specimens. In this study, we use this assay and a modification thereof to screen acute flaccid paralysis (AFP) samples that had previously tested negative for enteroviruses by the RD-L20B algorithm. Thirty paired stool suspensions collected in 2015 as part of the national AFP surveillance program in different states of Nigeria were analyzed in this study. The samples had previously tested negative for enteroviruses in the polio laboratory in accordance with the WHO-recommended RD-L20B-cellculture- based algorithm. Two samples that had previously been found to contain enteroviruses were included as positive controls. All samples were subjected to RNA extraction, the RT-snPCR assay and a modified version of the RT-snPCR. All amplicons were sequenced, and enteroviruses were identified using the enterovirus genotyping tool and phylogenetic analysis. Amplicons were recovered from the two controls and 50% (15/30) of the samples screened. Fourteen were successfully typed, of which, 7.1% (1/14), 21.4% (3/14), 64.3% (9/14) and 7.1% (1/14) were enterovirus (EV) -A, EV-B, EV-C and a mixture of EV-B and C (EV-C99 and E25), respectively. The two controls were identified as EV-C99 and coxsackievirus (CV) -A1, both of which belong to the species Enterovirus C. In one sample, poliovirus serotype 2 was detected and found to have the VP1 ILE143 variation and was therefore identified as a vaccine strain. The results of this study showed that significant proportion of enterovirus infections (including some with Sabin PV2) are being missed by the RD-L20B-cell-culture-based algorithm, thus highlighting the value of the RT-snPCR assay and its modifications. The circulation and preponderance of EV-C in Nigeria was also confirmed.Item Enterovirus A119 in A Child with Acute Flaccid Paralysis, Nigeria(MedCrave Group, 2016) Adeniji, J. A.; Oragwa, A. O.; George, U. E.; Ibok, U. I.; Faleye, T. O. C.; Adewumi, M. O.The oldest EV-A119 record was in 2008 in a chimpanzee in Cameroon and subsequently in more non-human primates and healthy children. Here we report for the first time the detection of EV-A119 in a child with Acute Flaccid Paralysis, thus suggesting possible association with a clinical condition in humans