Genetic characterization of parvoviruses identified in stray cats in Nigeria.

Parvoviruses are a major cause of haemorrhagic gastroenteritis, leukopenia and high mortality in cats and dogs. In this study, the presence and genetic characteristics of parvoviruses circulating among cats in Nigeria are reported. Faecal samples of stray cats from live animal markets in southwestern (Oyo and Osun States) and north-central (Kwara State) Nigeria were screened for the presence of parvoviral DNA using a qPCR. Positive samples were further characterized using a qPCR based on minor groove binder probes. Overall, 85/102 (83.3 %) stray cats tested positive for feline panleukopenia virus (FPV) DNA and one cat was co-infected with canine parvovirus-2 type a. Sequence analysis of the complete capsid region of 15 Nigerian FPV strains revealed that they were up to 99.9 % similar to the American reference strain FPV-b at the nucleotide level, and three of them presented amino acid mutations in key capsid residues. This is the first report of identification and molecular characterization of FPV strains in cats in Nigeria. The high prevalence of the virus emphasizes the need for constant surveillance of the circulation of parvoviruses in Nigeria and underscores the need to deploy an effective vaccination strategy.

Seromolecular surveillance of rabbit haemorrhagic disease virus in Nigeria.

Following the first 2020 rabbit haemorrhagic disease virus (RHDV) outbreak in Nigeria which caused massive mortalities in several rabbitries, there was a need to know the spread and strains circulating in the affected states. Over 100 rabbitries still existing post-RHDV outbreak in Ogun and Kwara States were investigated. A commercial enzyme-linked immunosorbent assay kit was used to screen for RHDV immunoglobulin G in 192 rabbit sera, while RHDV VP60 gene was amplified in RNA extracted from these sera and tissues (liver and/or spleen harvested from 37 carcasses necrotized) by reverse transcription-polymerase chain reaction (RT-PCR). Sequences obtained from the amplicons were subjected to phylogenetic analysis. The results revealed a seroprevalence of 82.3% (158/192). RHDV VP60 gene was detected in 15/17 (88.2%) and 2/20 (10.0%) carcasses from Ogun and Kwara States, respectively, while none of the sera was positive. Sequences of the two positive amplicons selected (one from each states) shared 98.95% nucleotide identity and belonged to RHDV 2/GI.2 strain. Also, nBLAST of these sequences revealed 98.43-99.55% homology with the prototype Nigerian RHDV strain RHDV/NGR/ILN/001 (MT996357.1). Furthermore, these strains clustered with this prototype and a German RHDV strain (LR899166.1). Pathologic lesions affecting the respiratory, cardiovascular, renal, lymphatic, and digestive systems were observed in necropsied carcasses. This study indicated that RHDV 2/GI.2 strain was the cause of 2020 RHD outbreak in Nigeria. Thus, while continuous public sensitization about RHD especially among rabbit farmers in Nigeria is important, efforts aimed at design and implementation of RHD vaccination policy, preferably using indigenous seed, should be expedited.

Serologic evidence of silent Rift Valley fever virus infection among occupationally exposed persons in northern Nigeria.

INTRODUCTION: Rift Valley fever (RVF) is a zoonotic disease caused by RVF virus (RVFV) and transmitted primarily by mosquitoes and contact with fluids and tissues of infected animals. First described in Kenya, it has spread to many African countries and beyond. In humans, it is sometimes misdiagnosed because the symptoms resemble those of influenza and/or malaria. Butchers, abattoir workers, and livestock keepers have the highest risk of infection. METHODOLOGY: In this study, serum samples collected between February and September 2019 from 196 individuals comprising of butchers (n = 121), abattoir/slaughterhouse workers (n = 55), and livestock keepers (n = 20) in Benue, Sokoto, and Borno States of northern Nigeria were screened using a commercial ELISA that detected anti-RVFV IgM and IgG alike (i.e., without discrimination). Data from administered questionnaires and the ELISA results were statistically analyzed. RESULTS: Thirty-nine (19.9%) of the 196 samples were positive for RVFV antibodies. The distribution by states showed that 17.4% (8/46), 21.7% (15/69), and 19.8% (16/81) of samples from Benue, Sokoto, and Borno States were seropositive, respectively. Additionally, 21.5% (26/121) butchers, 16.4% (9/55) abattoir workers, and 20% (4/20) livestock keepers were seropositive. CONCLUSIONS: These findings provide serological evidence for exposure of occupationally at-risk individuals in northern Nigeria to RVFV. The higher seropositivity obtained in Sokoto and Borno states could be due to contact of these individuals with infected animal blood/tissues, aborted fetuses, and unhindered transboundary movement of animals and animal products into these states which share international borders with Niger, Chad, and Cameroon where evidences of RVFV infections were recently reported.

Molecular detection of dugbe orthonairovirus in cattle and their infesting ticks (Amblyomma and Rhipicephalus (Boophilus)) in Nigeria.

Dugbe orthonairovirus (DUGV), a tick-borne zoonotic arbovirus, was first isolated in 1964 in Nigeria. For over four decades, no active surveillance was conducted to monitor the spread and genetic variation of DUGV. This study detected and genetically characterized DUGV circulating in cattle and their infesting ticks (Amblyomma and Rhipicephalus (Boophilus)) in Kwara State, North-Central Nigeria. Blood and or ticks were collected from 1051 cattle at 31 sampling sites (abattoirs and farms) across 10 local government areas of the State. DUGV detection was carried out by RT-qPCR, and positive samples sequenced and phylogenetically analysed. A total of 11824 ticks, mostly A. variegatum (36.0%) and R. (B.) microplus (63.9%), were obtained with mean tick burden of 12 ticks/cattle. Thirty-four (32 A. variegatum and two R. (B.) microplus) of 4644 examined ticks were DUGV-positive, whereas all of the cattle sera tested negative for DUGV genome. Whole genome sequence (S, M and L segments) and phylogenetic analyses indicate that the positive samples shared up to 99.88% nucleotide identity with and clustered around the Nigerian DUGV prototype strain IbAr 1792. Hence, DUGV with high similarity to the previously characterised strain has been detected in Nigeria. To our knowledge, this is the first report of DUGV in North-Central Nigeria and the most recent information after its last surveillance in 1974.

Identification of a Putative Novel Genotype of Avian Hepatitis E Virus from Apparently Healthy Chickens in Southwestern Nigeria.

Avian hepatitis E virus (aHEV) is the major etiological agent of hepatitis-splenomegaly syndrome (HSS), big liver and spleen disease (BLSD), and hepatic rupture hemorrhage syndrome (HRHS) in chickens. Infections with aHEV cause a significant decrease in egg production and increased mortality in chickens worldwide. However, studies on the prevalence of aHEV in Nigeria are scarce. In this study, serum (n = 88) and fecal samples (n = 110) obtained from apparently healthy layer chickens from three states in southwestern Nigeria were analyzed by nested reverse transcription-polymerase chain reaction (nRT-PCR) targeting the helicase and capsid gene for the presence of aHEV. Avian HEV was detected in 12.5% (n = 11/88) of serum samples and 9.1% (n = 10/110) of fecal samples tested. Phylogenetic analysis showed that five of the twelve identified aHEV sequences belonged to genotype 2. The remaining seven sequences were only distantly related to other known aHEV isolates. After amplification of the near-complete ORF2 fragment (1618 bp) and part of the ORF1 (582 bp) of isolate YF40_aHEV_NG phylogenetic analysis revealed a nucleotide sequence identity between 79.0 and 82.6% and 80.1 and 83.5%, respectively, to other known aHEV strains, indicating that the Nigerian isolate YF40_aHEV_NG belongs to a novel aHEV genotype. This is the first report of co-circulation of aHEV genotypes in chickens in Nigeria.

Serological evidence of avian HEV antibodies in apparently healthy chickens in southwest Nigeria.

Avian hepatitis E virus (aHEV) is associated with hepatitis-splenomegaly syndrome, big liver and spleen disease and hepatic rupture haemorrhage syndrome. However, the knowledge about aHEV in commercial layer chickens in Nigeria is scarce. In this study, 460 serum samples obtained from 36 apparently healthy commercial layer chicken flocks in three states (Ogun, Osun and Oyo States) of southwestern Nigeria were analysed by enzyme linked immunosorbent assay for the presence of anti-aHEV immunoglobulin Y (IgY) antibodies. In total, the overall seroprevalence of anti-aHEV antibodies was 14.6%. The serological analysis revealed that 75% of the flocks examined were positive for anti-aHEV IgY antibodies from chickens of various ages in all three states. The percentage of the seropositive chickens in the three states varied from flock to flock ranging from 60% to 88.8% and seropositive chickens were detected at any age (24-52 weeks of age) without significant differences between the age groups. This is the first report assessing the presence of aHEV antibodies in chickens from Nigeria. The detection of anti-aHEV antibodies in commercial layer chickens in this study emphasizes the importance of serosurveillance in disease monitoring due to the economic threat posed by aHEV as a result of decreased egg production and increased mortality in affected commercial layer chicken farms. However, further studies are essential to reveal the clinical implications and to assess the real burden of aHEV in Nigeria.

Real-time RT-PCR for COVID-19 diagnosis: challenges and prospects.

COVID-19 impacts global public health, economy, education, tourism/hospitality and sports; rapid and accurate testing of clinical samples dictate effective response. So far, the real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) is the assay of choice for COVID-19 diagnosis considering its rapidity and accuracy in informing on active coronavirus (CoV) infection. Presently, several RT-qPCR protocols with differing sensitivity/specificity are used for performing this assay; some of them are known to have generated debatable test results to constitute challenges worthy of consideration. This review provides a critical assessment of various published works on RT-qPCR assays used for COVID-19 diagnosis with their different indicators of positivity i.e., cycle threshold (Ct) cut-off values. Knowledge of diagnostic tests for COVID-19 is still evolving and, as a prospect, underscores the need for local validation of positive-negative Ct cut-off values when establishing RT-qPCR assays for SARS-CoV-2 detection.

ANTIBIOTIC RESISTANCE PROFILING AND MICROBIOTA OF THE UPPER RESPIRATORY TRACT OF APPARENTLY HEALTHY DOGS IN IBADAN, SOUTH WEST NIGERIA.

BACKGROUND: Rearing of dogs and other pets has become increasingly popular in modern society. Bacterial flora resides within the nasal and oral cavities of dogs and when chanced, can be pathogenic. Certain similarities between humans and dogs portends dangerous behavioral habits that could lead to zoonotic disease transmission. This study was aimed at isolation, identification and antibiotic profiling of bacteria from nasal swabs of apparently healthy dogs. The zoonotic risk was also considered. METHODOLOGY: A total of 173 nasal swabs were collected from 173 apparently healthy dogs. Structured questionnaires were administered to investigate human behavioral habits. RESULTS: Two hundred and twenty two (222) bacterial isolates were obtained from the culture with ten (10) potentially pathogenic bacteria in the order of Escherichia coli (18.5%), Proteus species (17.1%), Staphylococcus aureus (14.0%), Klebsiella species (9.0%), Acinetobacter species (9.0%), coagulase negative Staphylococcus species (7.7%), Pseudomonas species (6.8%), Actinobacter species (6.8%), Citrobacter species (5.9%) and Streptococcus species (5.4%). Overall, the Gram negative isolates showed resistance to ciprofloxacin (9.3%), sparfloxacin (16.0%), perfloxacin (17.3%), ofloxacin (21.6%), chloramphenicol (34..6%), gentamycin (36.4%), streptomycin (37.%), septrin (49.4%), amoxillin (59.3%), augmentin (62.3%) while the Gram positive bacteria showed resistance to ciprofloxacin (3.3%), perfloxacin (6.7%), erythromycin (13.3%), streptomycin (21.7%), rocephin (28.3%), septrin (28.3%), gentamycin (36.7%), zinnacef (68.3%), ampiclox (81.7%) and amoxillin (85.0%). Multi-drug resistance (MDR) to three or more antimicrobials was observed in some of the isolates. Seventy - seven resistance patterns were observed, 16 in Gram positive and 61 in Gram negative bacteria. CONCLUSION: This study revealed MDR to two or more antimicrobials in all the isolates. These can pose antibiotic resistance challenges in situation of primary or secondary canine respiratory infections. Also, this study revealed that 82% of the dog owners/lovers had less than 50cm face-to-face contact with these dogs while playing with them, thus increasing their chances of acquiring MDR bacteria from apparently healthy dogs.

FLOCK-BASED SURVEILLANCE FOR LOW PATHOGENIC AVIAN INFLUENZA VIRUS IN COMMERCIAL BREEDERS AND LAYERS, SOUTHWEST NIGERIA.

BACKGROUND: Flock surveillance systems for avian influenza (AI) virus play a critical role in countries where vaccination is not practiced so as to establish the epidemiological characteristics of AI needed for the development of prevention and control strategies in such countries. MATERIALS AND METHODS: As part of routine AI monitoring in southwest Nigeria, a competitive ELISA was used for detecting influenza A virus antibodies in the sera of 461 commercial breeder and layer birds obtained from different flocks in Oyo State, Nigeria while haemagglutination inhibiting antibodies against low pathogenic AI viruses (LPAIVs) were detected using H5N2, H7N7 and H9N2 subtype-specific antigens. Suspensions prepared from cloacal swabs were tested for AI virus RNA using reverse transcriptase-polymerase chain reaction. RESULTS: Results showed that influenza A virus antibody prevalence was 12.8% and 9.3% for breeders and layers, respectively while HI assay revealed 22.0%, 2.0% and 78.0% prevalence of LPAIV H5N2, H7N7 and H9N2 antibodies respectively. All cloacal swab suspensions were negative for AIV RNA. CONCLUSION: Since LPAI infections result in decreased or complete cessation of egg production in breeder and layer birds, increased infection severity due to co-infection with other poultry viruses have occasionally been transmitted to humans, the detection of LPAIV H5N2, H7N7 and H9N2 antibodies in these birds is of both economic and public health significance. These findings underscore the need for continuous flock monitoring as part of early warning measure to facilitate rapid detection and sustainable control of AI in Nigerian poultry.

Analysis of Culex and Aedes mosquitoes in southwestern Nigeria revealed no West Nile virus activity.

INTRODUCTION: Amplification and transmission of West Nile virus (WNV) by mosquitoes are driven by presence and number of viraemic/susceptible avian hosts. METHODS: In order to predict risk of WNV infection to humans, we collected mosquitoes from horse stables in Lagos and Ibadan, southwestern Nigeria. The mosquitoes were sorted and tested in pools with real-time RT-PCR to detect WNV (or flavivirus) RNA using WNV-specific primers and probes, as well as, pan-flavivirus-specific primers in two-step real-time RT-PCR. Minimum infection rate (MIR) was used to estimate mosquito infection rate. RESULTS: Only two genera of mosquitoes were caught (Culex, 98.9% and Aedes, 1.0%) totalling 4,112 females. None of the 424 mosquito pools tested was positive for WNV RNA; consequently the MIR was zero. Sequencing and BLAST analysis of amplicons detected in pan-flavivirus primer-mediated RT-PCR gave a consensus sequence of 28S rRNA of Culex quinquefasciatus suggesting integration of flaviviral RNA into mosquito genome. CONCLUSION: While the latter finding requires further investigation, we conclude there was little or no risk of human infection with WNV in the study areas during sampling. There was predominance of Culex mosquito, a competent WNV vector, around horse stables in the study areas. However, mosquito surveillance needs to continue for prompt detection of WNV activity in mosquitoes.

Serological Investigation of Akabane Virus Infection in Cattle and Sheep in Nigeria.

Akabane virus (AKAV) is recognized as an important pathogen that causes abortions and congenital malformations in ruminants. However, it has not received adequate attention in Nigeria. Therefore, in investigating this disease, serum samples from 184 (abattoir and farm) head of cattle and 184 intensively reared sheep from two states in southwest Nigeria were screened for antibodies against AKAV using enzyme-linked immunosorbent assay. An overall seropositivity of 70.1% (129/184) was obtained with antibodies being detectable in 73.8% of abattoir (trade) cattle and 40.0% in farm cattle, while 4.3% (8/184) seropositivity was observed in sheep. All the age groups of cattle tested had seropositive animals, 0-1 year (1/7, 14.3%), 2-3 years (17/34, 50.0%), 4-5 years (92/121, 76.0%), and >5 years (19/22, 86.4%), while in sheep only the age groups of 2-3 and 4-5 years showed seropositivity of 4.1% (4/97) and 8.2% (4/49), respectively. The detection of antibody-positive animals among unvaccinated cattle and sheep provides evidence of AKAV infection in Nigeria. These findings call for continuous monitoring of the disease among ruminants in order to ascertain the actual burden and increase awareness of the disease. This will facilitate early detection and aid the development of appropriate control measures against the disease in Nigeria.

Serological Survey for Avian Influenza in Turkeys in Three States of Southwest Nigeria.

Since the first outbreak of avian influenza (AI) in Nigeria in 2006, there has been continuous monitoring of the disease in chickens with little attention given to turkeys. As part of on-going surveillance for AI in southwest Nigeria, we used a competitive ELISA to detect anti-AI virus antibodies in 520 turkey sera obtained from poultry farms in Oyo, Osun, and Ondo states while haemagglutination inhibiting antibodies against low pathogenic AI viruses (LPAIVs) were detected using H3N8 and H5N2 subtype-specific antigens. The overall seroprevalence obtained by ELISA was 4.4% (23/520). Of the 23 ELISA-positive samples, 18 were positive for anti-AIV H3N8 antibodies only and four were positive for both anti-AIV H3N8 and H5N2 antibodies indicating a mixed infection, while five were negative for antibodies to either of the two AIV subtypes. Considering that turkeys have been implicated as a mixing vessel for generating influenza virus reassortants of human and avian origin, the detection of antibodies to LPAIV H3N8 and H5N2 in these turkeys is of public health concern. We advocate further studies to determine the potential role of turkeys in the zoonotic transmission of AIVs in Nigeria. Additionally, the practice of rearing turkeys with chickens should be discouraged.

High seroprevelance of West Nile virus antibodies observed in horses from southwestern Nigeria.

To investigate exposure of Nigerian horses to West Nile virus (WNV), we determined the seroprevalence rate of anti-WNV antibody in a cohort of 145 horses. Serum samples were collected from three locations in southwestern Nigeria between October, 2011, and July, 2012. The horses were asymptomatic and unvaccinated against WNV at the time of sampling. All sera were tested using a competition enzyme-linked immmunosorbent assay (ELISA) and by an immunoglobulin M (IgM)-specific ELISA. High rates of anti-WNV antibody prevalence were observed in all locations with a mean level of 90.3% (95% confidence interval 84.3-94.6%). None of the horses had detectable anti-WNV IgM. This is the first ELISA-based report of WNV seroprevalence in Nigerian horses and suggests that WNV is enzootic in the study areas, indicating a potential risk of infection in humans and animals.

Occurrence of Newcastle Disease and Infectious Bursal Disease Virus Antibodies in Double-Spurred Francolins in Nigeria.

The double-spurred francolin Francolinus bicalcaratus has been identified as a good candidate for future domestication due to the universal acceptability of its meat and its adaptability to anthropogenically altered environments. Therefore, in investigating the diseases to which they are susceptible, serum samples from 56 francolins in a major live-bird market (LBM) in Ibadan, southwestern Nigeria, were screened for antibodies against Newcastle disease (ND) and infectious bursal disease (IBD) viruses. Haemagglutination inhibition (HI) test and enzyme-linked immunosorbent assay (ELISA) revealed 25.0% and 35.7% prevalence of ND virus (NDV) antibodies, respectively, while 5.4% and 57.1% prevalence of IBD virus (IBDV) antibodies was detected by agar gel precipitation test (AGPT) and ELISA, respectively. This first report on the occurrence of NDV and IBDV antibodies in apparently healthy, unvaccinated double-spurred francolins from a LBM suggests that they were subclinically infected with either field or vaccine viruses and could thus serve as possible reservoirs of these viruses to domestic poultry. Furthermore, if they are to be domesticated for intensive rearing, a vaccination plan including ND and IBD should be developed and implemented.