Prevalence of vaccine-derived poliovirus in sewage waters in Maiduguri, Borno State, Nigeria.

After a long global battle with wild poliovirus, the virus has been defeated through researches and vaccination using the oral polio vaccine and inactivated polio vaccine as well as sensitization. The issue that is now of global concern is that of vaccine-derived poliovirus which emerged from the unstable oral polio vaccine. Ninety sewage water samples were collected from slums in Maiduguri using grab method, concentrated using two phase separation method and subjected to intratypic differentiation and vaccine-derived poliovirus screening. The result revealed the presence of Sabin 1in 17 samples (61.0%) and Sabin 3 in 22 samples (79.0%), all of which were positive after vaccine-derived poliovirus screening. The presence of strains of Sabin 1 and Sabin 3 in the sewage water samples collected is an indication of virus shedding in individuals which could be as a result of vaccination or contact with the faeces infected or vaccinated individuals.

Cryptic Zika virus infections unmasked from suspected malaria cases in Northeastern Nigeria.

INTRODUCTION: Although environmental and human behavioral factors in countries with Zika virus (ZIKV) outbreaks are also common in Nigeria, such an outbreak has not yet been reported probably due to misdiagnosis. The atypical symptoms of malaria and ZIKV infections at the initial phase could leverage their misdiagnosis. This study randomly recruited 496 malaria-suspected patients who visited selected health institutions in Adamawa, Bauchi, and Borno states for malaria tests. These patients' sera were analyzed for ZIKV antibodies using ELISA and plaque reduction neutralization tests (PRNT) at 90% endpoint. About 13.8% of Zika virus-neutralizing antibodies (nAb) did not cross-react with dengue, yellow fever, and West Nile viruses suggesting possible monotypic infections. However, 86% of the sera with ZIKV nAb also neutralized other related viruses at varied degrees: dengue viruses (60.7%), West Nile viruses (23.2%), yellow fever virus (7.1%) and 39.3% were co-infections with chikungunya viruses. Notably, the cross-reactions could also reflect co-infections as these viruses are also endemic in the country. The serum dilution that neutralized 90-100% ZIKV infectivity ranged from 1:8 to 1:128. Also, our findings suggest distinct protection against the ZIKV between different collection sites studied. As indicated by nAb, acute ZIKV infection was detected in 1.7% of IgM-positive patients while past infections occurred in 8.5% of IgM-negatives in the three states. In Borno State, 9.4% of IgG neutralized ZIKV denoting past infections while 13.5% were non-neutralizing IgM and IgG indicating other related virus infections. The age, gender, and occupation of the patients and ZIKV nAb were not significantly different. ZIKV nAb from samples collected within 1-7 days after the onset of symptoms was not significantly different from those of 7-10 days. A wider interval with the same techniques in this study may probably give better diagnostic outcomes. ZIKV nAb was significantly distinct among recipients and non-recipients of antibiotic/antimalaria treatments before seeking malaria tests. The inhibiting effect of these drugs on ZIKV infection progression may probably contribute to the absence of neurological disorders associated with the virus despite being endemic in the environment for several decades. Also, protection against ZIKV as marked by the nAb was different among the vaccinated and unvaccinated YF vaccine recipients. Thus, the YF vaccine may be a good alternative to the Zika vaccine in resource-constrained countries. CONCLUSION: The cryptic ZIKV infections underscore the need for differential diagnosis of malaria-suspected febrile patients for arboviruses, especially the Zika virus. The absence of systemic surveillance for the virus is worrisome because of its association with neurological disorders in newborns. Co-infections with other arboviruses may impact adversely on the management of these diseases individually.

Nearly Complete Genome Sequences of Type 2 Sabin-Like Polioviruses from Northern Nigerian Poliovirus Surveillance, 2016 to 2018.

We sequenced 109 type 2 Sabin-like poliovirus isolates that had been collected from acute flaccid paralysis patients or healthy children in Nigeria. Understanding the genetic makeup of these viruses may contribute to polio eradication efforts.

A year of genomic surveillance reveals how the SARS-CoV-2 pandemic unfolded in Africa.

The progression of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic in Africa has so far been heterogeneous, and the full impact is not yet well understood. In this study, we describe the genomic epidemiology using a dataset of 8746 genomes from 33 African countries and two overseas territories. We show that the epidemics in most countries were initiated by importations predominantly from Europe, which diminished after the early introduction of international travel restrictions. As the pandemic progressed, ongoing transmission in many countries and increasing mobility led to the emergence and spread within the continent of many variants of concern and interest, such as B.1.351, B.1.525, A.23.1, and C.1.1. Although distorted by low sampling numbers and blind spots, the findings highlight that Africa must not be left behind in the global pandemic response, otherwise it could become a source for new variants.

Diagnostic performance of a colorimetric RT -LAMP for the identification of SARS-CoV-2: A multicenter prospective clinical evaluation in sub-Saharan Africa.

BACKGROUND: Management and control of the COVID-19 pandemic caused by the severe acute respiratory syndrome coronavirus SARS-CoV-2 is critically dependent on quick and reliable identification of the virus in clinical specimens. Detection of viral RNA by a colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a simple, reliable and cost-effective assay, deployable in resource-limited settings (RLS). Our objective was to evaluate the intrinsic and extrinsic performances of RT-LAMP in RLS. METHODS: This is a multicenter prospective observational study of diagnostic accuracy, conducted from October 2020 to February 2021 in four African Countries: Cameroon, Ethiopia, Kenya and Nigeria; and in Italy. We enroled 1657 individuals who were either COVID-19 suspect cases, or asymptomatic and presented for screening. RNA extracted from pharyngeal swabs was tested in parallel by a colorimetric RT-LAMP and by a standard real time polymerase chain reaction (RT-PCR). FINDINGS: The sensitivity and specificity of index RT LAMP compared to standard RT-PCR on 1657 prospective specimens from infected individuals was determined. For a subset of 1292 specimens, which underwent exactly the same procedures in different countries, we obtained very high specificity (98%) and positive predictive value (PPV = 99%), while the sensitivity was 87%, with a negative predictive value NPV = 70%, Stratification of RT-PCR data showed superior sensitivity achieved with an RT-PCR cycle threshold (Ct) below 35 (97%), which decreased to 60% above 35. INTERPRETATION: In this field trial, RT-LAMP appears to be a reliable assay, comparable to RT-PCR, particularly with medium-high viral loads (Ct < 35). Hence, RT-LAMP can be deployed in RLS for timely management and prevention of COVID-19, without compromising the quality of output.

Assessment of immunity against Yellow Fever virus infections in northeastern Nigeria using three serological assays.

Poor systematic surveillance for Yellow Fever virus (YFV) is primarily due to lack of affordable diagnostic facilities in resource-constrained countries. This study aimed at providing evidence-based information on immunity against Yellow Fever with a view to assessing the possibility of the recent epidemics persisting in Nigeria. Six hundred patients with febrile illness seeking malaria test in selected hospitals were tested for YFV antibody using three serological assays: ELISA IgM, microneutralization test (MNT) and plaque reduction neutralization test (PRNT). The three assays commonly detected YFV antibody (Ab) in 1.7% patients, MNT: IgM in 8.3%, IgM: PRNT in 7.1%, and MNT: PRNT in 3.2%. Immunity against YF was significantly higher in Bauchi and Borno than Adamawa and children aged 0-9 years compared to 20-29 years. YFV neutralizing antibody (nAb) strongly correlated with the vaccination status of the patients. More unvaccinated patients had nAb compared with the vaccinated. Immunity against YF among treated patients with antibiotic and/or antimalaria before sample collection inversely correlated with the untreated. YVnAb among unvaccinated indicates natural infections. Acute YFV infections were mistaken for malaria and natural infections are ongoing. Individuals aged more than or equal to 20 years should be targeted during mass vaccination campaigns. With low population immunity, repetitive YF epidemics in Nigeria is not yet over. The current policy on Yellow Fever vaccination in Nigeria still leaves a large unimmunized population at the risk of epidemics. Sufficient mass vaccination in combination with National Programme on Immunization remains key to averting YF epidemics.

Distribution pattern and prevalence of West Nile virus infection in Nigeria from 1950 to 2020: a systematic review.

OBJECTIVES: West Nile virus (WNV) is a re-emerging mosquito-borne viral infection. This study investigated the pooled prevalence pattern and risk factors of WNV infection among humans and animals in Nigeria. METHODS: A systematic review was conducted of eligible studies published in PubMed, Scopus, Google Scholar, and Web of Science from January 1, 1950 to August 30, 2020. Peer-reviewed cross-sectional studies describing WNV infections in humans and animals were systematically reviewed. Heterogeneity was assessed using the Cochrane Q statistic. RESULTS: Eighteen out of 432 available search output were eligible and included for this study. Of which 13 and 5 were WNV studies on humans and animals, respectively. Although 61.5% of the human studies had a low risk of bias, they all had high heterogeneity. The South West geopolitical zone of Nigeria had the highest pooled prevalence of anti-WNV immunoglobulin M (IgM; 7.8% in humans). The pooled seroprevalence of anti-WNV IgM and immunoglobulin G (IgG) was 7.1% (95% confidence interval [CI], 5.9 to 8.3) and 76.5% (95% CI, 74.0 to 78.8), respectively. The WNV RNA prevalence was 1.9% (95% CI, 1.4 to 2.9), while 14.3% (95% CI, 12.9 to 15.8) had WNV-neutralizing antibodies. In animals, the pooled seroprevalence of anti-WNV IgM and IgG was 90.3% (95% CI, 84.3 to 94.6) and 3.5% (95% CI, 1.9 to 5.8), respectively, while 20.0% (95% CI, 12.9 to 21.4) had WNV-neutralizing antibodies. Age (odds ratio [OR], 3.73; 95% CI, 1.87 to 7.45; p<0.001) and level of education (no formal education: OR, 4.31; 95% CI, 1.08 to 17.2; p<0.05; primary: OR, 7.29; 95% CI, 1.80 to 29.6; p<0.01) were significant risk factors for WNV IgM seropositivity in humans. CONCLUSIONS: The findings of this study highlight the endemicity of WNV in animals and humans in Nigeria and underscore the need for the One Health prevention and control approach.

Leucocytes and Th-associated Cytokine Profile of HIV-Leishmaniasis Co-Infected Persons Attending Abuja Teaching Hospital, Nigeria.

OBJECTIVE: T-helper cells (Th)-1& -2 cytokines homeostasis control or predict clinical outcome of infected persons, especially those with HIV /AIDS. This case-control study evaluated the leucocytes differentials, TNF-alpha, interleukin (IL)-2 and -10 levels among HIV infected persons with serological evidence of leishmaniasis attending University of Abuja Teaching Hospital, Nigeria. MATERIALS AND METHODS: Blood samples from 28 HIV infected persons who had Leishmania donovani rK39 and Immunoglobulin-G (IgG) positive (group 1), 30 age- & -sex matched HIV infected persons without Leishmania antibodies (group 2) and 30 apparently healthy persons without HIV and Leishmania antibodies (group 3). Full blood counts, TNF alpha, IL-2 and -10 levels were analyzed using automated hematology analyzer and ELISA, respectively. Structured questionnaires were used to collate biodata and clinical presentations of participants. RESULTS: Ten (35.7%) participants in group 1 were on ART, 15 (50%) in group 2 were on ART, while group 3 were ART naïve. There were significantly higher values in basophil (4.4±2.5%) and eosinophil counts (12.9±3.8%) in HIV/leishmania coinfected persons (p<0.005). However, other white cells subpopulation was significantly lower in HIV/leishmania co-infected participants (p<0.05). There was significantly reduced CD4+ T cell counts ([119±26 versus 348±63 versus 605±116 cells/mm3]), TNF-alpha ([36.82±8.21 versus 64.67±12.54 versus 254.98±65.59 pg/mL]) and IL-2 levels ([142.14±20.91 versus 507.6±84.42 versus 486.62±167.87 pg/mL]) among HIV/Leishmania co-infected participants compared to group 2 and group 3 participants, respectively. However, higher IL-10 level (80.35±14.57 pg/mL) was found in HIV/Leishmania co-infected participants as opposed to the HIV monoinfected (62.2±10.43 pg/mL) and apparently healthy persons (23.97±4.88 pg/mL) (p<0.001). CONCLUSION: Eosinophil, basophil counts and serum IL-10 level were high in HIV/Leishmania coinfected persons, demonstrating parasite-induced hypersensitivity and immunosuppression.

Implications of SARS-CoV-2 genetic diversity and mutations on pathogenicity of the COVID-19 and biomedical interventions.

OBJECTIVE: Coronavirus disease 2019 (COVID-19) has caused an unprecedented global health emergency. The COVID-19 pandemic has claimed over 350,000 human lives within five months of its emergence, especially in the USA and the European continent. This study analysed the implications of the genetic diversity and mutations in SARS-CoV-2 on its virulence diversity and investigated how these factors could affect the successful development and application of antiviral chemotherapy and serodiagnostic test kits, and vaccination. METHODS: All the suitable and eligible full text articles published between 31st December 2019 and 31st May 2020 were filtered and extracted from "PubMed", "Scopus", "Web of Science", and "Hinari" and were critically reviewed. We used the Medical Subject Headings (MeSH) terms "COVID-19, "Mutation", "Genetic diversity", "SARS-CoV-2", "Virulence", "Pathogenicity", "Evolution" and "SARS-CoV-2 transmission" for this search. RESULTS: Our search showed that SARS-CoV-2 has persistently undergone significant mutations in various parts of its non-structural proteins (NSPs) especially NSP2 and NSP3, S protein, and RNA-dependent RNA polymerase (RdRp). In particular, the S protein was found to be the key determinant of evolution, transmission, and virulence of SARS-CoV-2, and could be a potential target for vaccine development. Additionally, RdRp could be a major target in the development of antivirals for the treatment of COVID-19. CONCLUSION: Given the critical importance of mutations in the pathogenicity of SARS-CoV-2 and in the development of sero-diagnostics, antivirals, and vaccines, this study recommends continuous molecular surveillance of SARS-CoV-2. This approach would potentially prompt identification of new mutants and their impact on ongoing biomedical interventions and COVID-19 control measures.

Prevalence of locally undetected acute infections of Flaviviruses in North-Eastern Nigeria.

The burden of Arboviral infections is largely underestimated in Africa, particularly in North-Eastern Nigeria. A total of 200 serum samples were collected from patients exhibiting febrile illness who visited the State Specialist Hospital in Maiduguri for medical attention between March and April 2018. Sera were tested for Flavivirus RNA by a pan-flaviviral primer set using hemi-nested RT PCR. Twenty-six samples were positive for flaviviral RNA and sequence analysis indicated a high number of West Nile virus infections and one case of Zika virus. In-house recombinant NS1-based IgM ELISA indicated 47 % of WNV and 22 % of ZIKV infections. These data were also compared to commercially available assays for West Nile and Zika viruses. Finally, NS1 IgG ELISA was conducted for Dengue, Zika, West Nile and Usutu viruses. For serum samples detected by at least one flavivirus, 945% tested positive by NS1 IgG antibodies, while only 5.5 % of the patients were negative for all. To conclude, there is a high prevalence rate of arbovirus infections in the region, including Zika and Usutu viruses that were not previously detected. Interestingly, the analysis was conducted using in-house tools to allow the implementation of a sustainable surveillance protocol locally.

Prevalence Pattern of Chikungunya Virus Infection in Nigeria: A Four Decade Systematic Review and Meta-analysis.

Chikungunya (CHIK) is a re-emerging and myo-arthritogenic arboviral infection that has affected significant global population. However, CHIK is a neglected disease in Nigeria. This study aimed to estimate the pooled prevalence pattern of CHIK virus infection in Nigeria. A systematic review of eligible articles was conducted from "PubMed", "Scopus", "Google Scholar" and "Web of Science", between January 1980 to February 2020. Peer-reviewed articles describing CHIKV infection in cross-sectional studies were systematically reviewed. Random-effect model was used to pool the prevalence of CHIKV infection and associated sociodemographic data reported from eligible studies. In total, there were 10 published articles on CHIKV infection. Of these, 7 were cross-sectional studies, which comprised of 1347 pooled participants. The pooled anti-CHIKV IgM and IgG seroprevalence were 26.7% (95% CI: 23.2 - 30.4) and 29.3% (95% CI: 26.2 -32.6), respectively.  Of the pooled studies, there were 3.8% (95% CI: 2.0-6.4) CHIKV RNA positive cases and 46.1% prevalence of CHIKV neutralizing antibodies. Of the 6 geopolitical zones in Nigeria, Northeast had the highest serological evidence of CHIKV infection. There was a significance association between the prevalence of anti-CHIKV and geopolitical zones of Nigeria (χ²= 70.04; p˂0.0001). Sex (p ˂0.0001; OR= 1.87 [1.47 - 2.38]) and level of education (p ˂0.0001; OR= 2.74 [1.89 - 3.95]) were significant risk factors for pooled anti-CHIKV IgM seropositivity. However, no significant association was found with other sociodemographic variables (p ˃0.05). Although there was paucity of data on CHIKV research in Nigeria, this meta-analysis revealed a high prevalence of CHIKV infection in the country.

Molecular characterization of group A rotavirus strains detected in children with diarrhea admitted to Nigerian hospitals in 2013.

Group A rotaviruses (RVA) are the leading cause of acute gastroenteritis (AGE) in children worldwide and cause up to 455,000 deaths annually, mostly in developing countries. During 2013, 66 RVAs from children with AGE admitted to four Nigerian hospitals were investigated. The G3P[6], G1P[8] and G2P[4] genotypes predominated. The VP7 and/or VP4 genes of 18 G3P[6]/[8]/[4], six G2P[4], three G12P[8]/[4], and two G1P[8] RVA strains were sequenced. The G3P[6] strains belonged to lineage G3-III and were different from G3 strains widespread in Asia. Phylogenetic analysis revealed substantial sequence conservation, suggesting continuing evolution and genomic reassortment but no zoonotic RVA transmission from animals.

Evidence of arbovirus co-infection in suspected febrile malaria and typhoid patients in Nigeria.

INTRODUCTION: Clinical symptoms of malaria and typhoid infections are virtually indistinguishable from those initially seen in many arbovirus infections. Here we describe arbovirus co-infection detected in 310 sera samples collected from febrile, clinically suspected malaria/typhoid patients in Borno State, Nigeria. METHODOLOGY: Tested initially for Plasmodium falciparum by microscopy and for Salmonella Typhi by Widal test, samples were subsequently tested for chikungunya (CHIKV), yellow fever (YFV), dengue (DENV) and West Nile viruses (WNV) by plaque reduction neutralization test. RESULTS: While 92% of patients tested positive for malaria, typhoid, an arbovirus infection, or a combination of one or more of these types of infections, less than 1% of the patients tested positive for malaria alone and only 3.9% tested positive for typhoid alone.  Approximately half of the patients tested positive for infection with a single arbovirus (48%) regardless of the presence or absence of malaria or typhoid. Of those who showed 90% to 95% virus neutralization, 67.7% had neutralizing antibodies against DENV, 50% against CHIKV, 25% against WNV and 8.7% against YFV. Eight per cent tested negative against all six pathogens, suggesting that other arboviruses not tested for in this study may also be circulating in Nigeria. CONCLUSIONS: The results suggest that misdiagnosis of arbovirus co-infections as malaria infections, combined with a lack of virus surveillance and underreporting of arbovirus infections, increases the potential for undetected and uncontrolled spread of important vector-borne arboviruses becoming serious underlying public health concerns in Nigeria.

High variety of known and new RNA and DNA viruses of diverse origins in untreated sewage.

Deep sequencing of untreated sewage provides an opportunity to monitor enteric infections in large populations and for high-throughput viral discovery. A metagenomics analysis of purified viral particles in untreated sewage from the United States (San Francisco, CA), Nigeria (Maiduguri), Thailand (Bangkok), and Nepal (Kathmandu) revealed sequences related to 29 eukaryotic viral families infecting vertebrates, invertebrates, and plants (BLASTx E score, <10(-4)), including known pathogens (>90% protein identities) in numerous viral families infecting humans (Adenoviridae, Astroviridae, Caliciviridae, Hepeviridae, Parvoviridae, Picornaviridae, Picobirnaviridae, and Reoviridae), plants (Alphaflexiviridae, Betaflexiviridae, Partitiviridae, Sobemovirus, Secoviridae, Tombusviridae, Tymoviridae, Virgaviridae), and insects (Dicistroviridae, Nodaviridae, and Parvoviridae). The full and partial genomes of a novel kobuvirus, salivirus, and sapovirus are described. A novel astrovirus (casa astrovirus) basal to those infecting mammals and birds, potentially representing a third astrovirus genus, was partially characterized. Potential new genera and families of viruses distantly related to members of the single-stranded RNA picorna-like virus superfamily were genetically characterized and named Picalivirus, Secalivirus, Hepelivirus, Nedicistrovirus, Cadicistrovirus, and Niflavirus. Phylogenetic analysis placed these highly divergent genomes near the root of the picorna-like virus superfamily, with possible vertebrate, plant, or arthropod hosts inferred from nucleotide composition analysis. Circular DNA genomes distantly related to the plant-infecting Geminiviridae family were named Baminivirus, Nimivirus, and Niminivirus. These results highlight the utility of analyzing sewage to monitor shedding of viral pathogens and the high viral diversity found in this common pollutant and provide genetic information to facilitate future studies of these newly characterized viruses.

Possible cross-species transmission of circoviruses and cycloviruses among farm animals.

Circoviruses consist of highly prevalent and genetically diverse porcine and avian pathogens. The genomes of cycloviruses, a proposed new genus in the family Circoviridae, were recently identified in human and chimpanzee faeces. Here, six cyclovirus and four circovirus genomes from the tissues of chickens, goats, cows, and a bat were amplified and sequenced using rolling-circle amplification and inverse PCR. A goat cyclovirus was nearly identical to a cyclovirus from a cow. USA beef contained circoviruses with >99% similarity to porcine circovirus 2b. Circoviruses in chicken were related to those of pigeons. The close genetic similarity of a subset of cycloviruses and circoviruses replicating in distinct animal species may reflect recent cross-species transmissions. Further studies will be required to determine the impact of these highly prevalent infections on the health of farm animals.

Human bocaviruses are highly diverse, dispersed, recombination prone, and prevalent in enteric infections.

A new species of parvovirus, tentatively named human bocavirus 4 (HBoV4), was genetically characterized. Among 641 feces samples obtained from children and adults, the most commonly detected bocavirus species were, in descending order, HBoV2, HBoV3, HBoV4, and HBoV1, with an HBoV2 prevalence of 21% and 26% in Nigerian and Tunisian children, respectively. HBoV3 or HBoV4 species were found in 12 of 192 patients with non-polio acute flaccid paralysis in Tunisia and Nigeria and 0 of 96 healthy Tunisian contacts (P = .01). Evidence of extensive recombination at the NP1 and VP1 gene boundary between and within bocavirus species was found. The high degree of genetic diversity seen among the human bocaviruses found in feces specimens, relative to the highly homogeneous HBoV1, suggest that this worldwide-distributed respiratory pathogen may have recently evolved from an enteric bocavirus after acquiring an expanded tropism favoring the respiratory tract. Elucidating the possible role of the newly identified enteric bocaviruses in human diseases, including acute flaccid paralysis and diarrhea, will require further epidemiological studies.

A novel picornavirus associated with gastroenteritis.

A novel picornavirus genome was sequenced, showing 42.6%, 35.2%, and 44.6% of deduced amino acid identities corresponding to the P1, P2, and P3 regions, respectively, of the Aichi virus. Divergent strains of this new virus, which we named salivirus, were detected in 18 stool samples from Nigeria, Tunisia, Nepal, and the United States. A statistical association was seen between virus shedding and unexplained cases of gastroenteritis in Nepal (P = 0.0056). Viruses with approximately 90% nucleotide similarity, named klassevirus, were also recently reported in three cases of unexplained diarrhea from the United States and Australia and in sewage from Spain, reflecting a global distribution and supporting a pathogenic role for this new group of picornaviruses.