Factors associated with foot-and-mouth disease seroprevalence in small ruminants and identification of hot-spot areas in northern Nigeria.

Many small ruminants infected with foot-and-mouth disease (FMD) remain asymptomatic, with the capacity to promote silent viral spread within domestic and wildlife species. However, little is known about the epidemiological role played by small ruminants in FMD. In particular, there are few studies that examine FMD seroprevalence, spatial patterns and risk factors for exposure in small ruminants. A cross-sectional study was conducted in northern Nigeria (Bauchi, Kaduna, and Plateau States) to determine the true seroprevalence of FMD in backyard small ruminants, identify factors associated with FMD seroconversion at animal and household levels, and identify spatial patterns for FMD virus exposure. Data on animal (n = 1800) and household (n = 300) characteristics were collected using a standardised questionnaire. Sera samples from 1800 small ruminants were tested for antibodies against non-structural proteins of FMD virus. True seroprevalence was estimated stochastically to account for variability and uncertainty in the test sensitivity and specificity previously reported. Risk factors for FMD seropositivity were identified at animal and household levels and spatial patterns were determined. The overall true seroprevalence for FMD virus, in the small ruminant population tested, was estimated to be 10.2 % (95 % Credible Interval (CrI) 0.0-19.0), while State-level estimates were 17.3 % (95 % CrI 0.0-25.8) for Kaduna, 6.9 % (95% CrI 0.0-15.8) for Bauchi, and 3.6 % (95 % CrI 0.0-12.6) for Plateau. State and species were the main risk factors identified at animal level, with interaction detected between them. Compared to goats in Plateau, the odds of testing positive were higher for goats in Bauchi (Odds Ratio (OR)= 1.83, 95 % CI 1.13-2.97, p = 0.01) and Kaduna (OR=2.97, 95 % CI 1.89-4.67, p < 0.001), as well as for sheep in Plateau (OR=3.78, 95 % CI 2.08-6.87, p < 0.001), Bauchi (OR=1.61, 95 % CI 0.91-2.84, p = 0.10), and Kaduna (OR=3.11, 95 % CI 1.61-6.01, p = 0.001). Households located in Kaduna were more likely to have a higher number of seropositive SR compared to those in Plateau (Prevalence Ratio (PR)= 1.75, 95 % CI 1.30-2.36, p < 0.001), and households keeping sheep flocks were more likely to be seropositive (from 1 to 10 sheep: PR=1.39, 95 % CI 1.05-1.82, p = 0.02; more than 10 sheep: PR=1.55, 95 % CI 1.12-2.15, p = 0.008) compared to those that did not keep sheep. A hot-spot was detected in Kaduna, and a cold-spot in Plateau. These results reveal that small ruminants had been recently exposed to FMD virus with spatial heterogeneity across the study area.

A survey of contagious ecthyma and molecular characterization of Orf virus in sheep and goats in Nigeria (2014-2016).

Background: Outbreaks of contagious ecthyma (CE) are frequently reported in sheep and goat flocks in Nigeria with severe clinical outcomes. CE is a debilitating and economically important disease primarily affecting sheep and goats caused by the Orf virus (ORFV). Despite field reports of CE in the country, there is no concise country-wide epidemiological data on the disease and limited genetic data of circulating Nigerian ORFV are available in the public domain. Aim: An epidemiological survey of CE and molecular characterization of ORFV circulating in Nigeria from 2014 to 2016. Method: Data were collected using designed questionnaires, administered to veterinarians and farmers in selected States of Nigeria. Samples were collected during passive surveillance for CE from 2014 to 2016 which were analyzed by polymerase chain reaction (PCR). The A32L and B2L genes of circulating ORFV were also characterized. Results: Analysis of the questionnaire showed that 69.54% (n = 82/118) of the farmers claimed to have experienced CE in their flocks with average morbidity and mortality rates of 25% and 15%, respectively. A total of 113 veterinarians participated in the study, with 69.9% (n = 79) familiar with CE and claimed CE causes morbidity rates of 25%-37% and mortality rates of 10%-15% in sheep and goats. Laboratory results revealed that ORFV was detected in 72% (18/25) of outbreak samples analyzed by real-time PCR. Phylogenetic analysis of A32L and B2L genes revealed that Nigerian ORFV sequences belong to clusters I and II and are similar to viruses from India, Ethiopia, and China. Conclusions: This study is the first nationwide epidemiological data on the status of CE in sheep and goats in Nigeria. It is also the first report of molecular characterization of two genes of ORFV circulating and causing outbreaks in small ruminants in the country. This study showed that CE is under-reported, widespread and of economic importance to sheep and goat farmers in Nigeria.

A Retrospective Study of the Seroprevalence of Dengue Virus and Chikungunya Virus Exposures in Nigeria, 2010-2018.

Arboviruses are important public health threats in many regions of the world. Nigeria has experienced outbreaks of arboviruses over the past decades, leading to concerns of widespread endemicity, which are frequently misdiagnosed. This study aimed to determine the seroprevalence of dengue virus (DENV) (a flavivirus) and chikungunya virus (CHIKV) (an alphavirus) infections in three major population centers of Nigeria. A convenience sample of 701 sera was collected from both healthy and febrile participants between August 2010 and March 2018. Sera were tested for prior exposure to CHIKV virus and DENV using indirect IgG ELISA. Results showed that 54.1% (379/701) of participants were seropositive for anti-DENV antibodies, 41.3% (290/701) were seropositive for anti-CHIKV antibodies, and 20.1% (141/701) had previous exposure to both. The seropositivity for prior CHIKV exposure and prior exposure to DENV and CHIKV was significantly associated with age (CHIKV: OR = 2.7 (95% CI: 1.7-4.3); DENV and CHIKV: OR = 2.2 (95% CI: 1.2-4.0) for adults compared to participants under 18 years old). Overall, the high seropositivity across all age groups suggests that arboviral infections are prevalent in Nigeria and indicates that surveillance and further epidemiological studies are required to determine the true burden of these infections and the spectrum of diseases associated with these exposures.

Molecular Characterization of Southern African Territories 2 (SAT2) Serotype of Foot-and-Mouth Disease Virus from Nigeria in 2017 to 2018.

This report describes the nucleotide sequences of eight Southern African Territories 2 (SAT2) serotype foot-and-mouth disease virus strains from 2017 to 2018 outbreaks in cattle in Nigeria. These viruses belong to topotype VII of SAT2 and were closely related to previous isolates from Nigeria and other West African countries.

Complex Circulation of Foot-and-Mouth Disease Virus in Cattle in Nigeria.

Nigeria is a large densely populated country in West Africa. Most of its livestock is raised in a pastoralist production system with typical long distance migration in search of water and feed. As the demand for animal products largely exceeds the domestic production, large numbers of livestock are imported from neighboring countries without sanitary restrictions. In Nigeria, foot-and-mouth disease virus (FMDV) serotypes O, A, and Southern African Territories (SAT)2 are endemic for a long time. Clinical outbreaks of FMD due to serotype SAT1 are described again since 2015, after an absence of more than 30 years. Historically, outbreaks of FMD due to serotypes O, A, SAT1, and SAT2 were each time associated with trade of cattle entering Nigeria from neighboring countries. In the present study, tissue samples from 27 outbreaks of FMD were collected in Nigerian cattle from 2012 until 2017 in six different States and in the Federal Capital Territory. FMDV was isolated and serotyped and further characterized by VP1 sequencing and phylogenetic analysis to gain more knowledge on FMDV circulation in Nigeria. Half of the outbreaks were characterized as FMDV topotype O/EA-3, while outbreaks with other serotypes and topotypes were-in descending order-less prevalent: A/Africa/G-IV, SAT1/X, SAT2/VII, and O/WA. The high dynamics and omnipresence of FMD in Nigeria were illustrated in Plateau State where FMDV serotypes O, SAT1, and SAT2 were isolated during the course of the study, while at some point in the study, outbreaks due to FMDV serotype A were observed in three remote States. The genetic and phylogenetic analysis suggests a mixed origin of FMD outbreaks. Some outbreaks seem to be caused by sustained local transmission of FMDV strains present in Nigeria since a number of years, while other outbreaks seem to be related to recent incursions with new FMDV strains. The role of African buffaloes in the etiology of FMD in Nigeria is unclear, and sampling of wildlife is needed. The results of the present study suggest that systematic sample collection is essential to understand the complex concomitance of FMDV strains in Nigeria and essential to support the implementation of a vaccination-based control plan.

Characterization of transboundary foot-and-mouth disease viruses in Nigeria and Cameroon during 2016.

Continuous surveillance for foot-and-mouth disease (FMD) in endemic settings such as West Africa is imperative to support improved local and regional control plans, with the long-term goal of regional eradication. This paper describes the genetic characterization of FMD viruses (FMDV) obtained from outbreaks in Nigeria (n = 45) and Cameroon (n = 15) during 2016 and from archival samples (n = 3) retrieved from a 2014 outbreak in Nigeria. These viruses were analysed in the context of previously published FMDV sequences from the region. Four FMDV serotypes: O, A, SAT1 and SAT2, were detected. Phylogenetic analyses of the VP1 coding sequences indicate the continuity of FMDV serotype O East Africa-3 (O/EA-3), serotype A AFRICA genotype G-IV (A/AFRICA/G-IV) and serotype South African Territories (SAT) 2 lineage VII (SAT2/VII). The FMDV SAT1 topotype X (SAT1/X), which emerged in Nigeria in 2015, continued to be associated with outbreaks in the region during 2016, and SAT1 is reported for the first time from Cameroon. Additionally, a re-emergence or re-introduction of the serotype O West Africa (O/WA) topotype in Nigeria is described herein. Our findings indicate a consistent, pan-serotypic relationship between FMDV strains detected in Cameroon and Nigeria. Additionally, FMDV strains from West Africa obtained in this study were genetically related to those occurring in East and North Africa. These phylogenetic relationships suggest that animal movements (pastoralism and/or trade) are important factors for virus spread across the African continent. These data provide critical baselines which are a necessary component of Stages 0 and 1 of the Progressive Control Pathway of FMD (PCP-FMD). Specifically, characterizing the existing virus strains (risk) provides the basis for the comprehensive risk-based control plan which is the requisite criteria for Nigeria's transition to Stage 2 of PCP-FMD, and for coordinated regional control of FMD.

Participatory survey of Rift Valley fever in nomadic pastoral communities of North-central Nigeria: The associated risk pathways and factors.

BACKGROUND: Rift Valley fever (RVF) is an emerging neglected mosquito-borne viral zoonotic disease of domestic animals and humans, with potential for global expansion. The objectives of this study were: to assess perceived relative burden and seasonality of RVF in nomadic cattle herds and validate the burden with sero-prevalence impact; and assess perceived risk factors associated with the disease and risk pathways for RVF virus in nomadic pastoral herds of North-central Nigeria using pastoralists' existing veterinary knowledge. METHODS: Participatory Epidemiology (PE) survey was conducted in Fulani nomadic pastoral communities domiciled in Niger State between January and December 2015. A cross-sectional sero-prevalence investigation was also carried out in nomadic pastoral cattle herds to validate outcomes of PE. A total of nine nomadic pastoral communities were purposively selected for qualitative impact assessment using Participatory Rural Appraisal tools, while 97 cattle randomly sampled from 15 purposively selected nomadic herds and had their sera analyzed using c-ELISA. Kendall's Coefficient of Concordance W statistics and OpenEpi 2.3.1 were used for statistical analyses. RESULTS: Mean proportional piles (relative burden) of RVF (Gabi-gabiF) was 8.3%, and nomads agreement on the burden was strong (W = 0.6855) and statistically significant (P<0.001). This was validated by 11.3% (11/97; 95% CI: 6.1-18.9) sero-positivity (quantitative impact). Mean matrix scores of prominent clinical signs associated with RVF were fever (3.1), anorexia (2.1), abortion (4.1), nasal discharge (3.3), neurological disorder (8.4), diarrhoea (3.2), and sudden death (4.4), with strong agreement (W = 0.6687) and statistically significant (p<0.001). Mean proportional piles of pastoralists' perceived risk factors identified to influenced RVF occurrence were: availability of mosquitoes (18 piles, 17.6%), high cattle density (16 piles, 15.9%) and high rainfall (12 piles, 12.2%). Agreement on the risk factors was strong (W = 0.8372) and statistically significant (p<0.01). Mean matrix scores for the Entry pathway of RVF virus into the nomadic pastoral herds were: presence of RVFV infected mosquitoes (tiny biting flies) (7.9), presence of infected cattle in herds (8.4), and contacts of herd with infected wild animals at grazing (10.1). Mean matrix scores for the Spread pathway of RVF virus in herds were bites of infected mosquitoes (5.1), contacts with infected aborted fetuses/fluids (7.8), and contaminated pasture with aborted fetuses/fluids (9.7). Agreement on risk pathways was strong (W = 0.6922) and statistically significant (p<0.03). Key informants scored RVF to occurred more in Damina or late rainy season (5.3), followed by Kaka or early dry season (3.3), with strong agreement (W = 0.8719) and statistically significant (P<0.01). This study highlighted the significant existing knowledge level about RVF contained in nomadic pastoralists. CONCLUSIONS: The use of PE approach is needful in active surveillance of livestock diseases in pastoral communities domiciled in highly remote areas. RVF surveillance system, control and prevention programmes that take the identified risk factors and pathways into consideration will be beneficial to the livestock industry in Nigeria, and indeed Africa. An 'OneHealth' approach is needed to improve efficiency of RVF research, surveillance, prevention and control systems, so as to assure food security and public health in developing countries.

Complete Genome Sequences of Four Foot-and-Mouth Disease Viruses of Serotype South African Territories 1 (SAT 1), Topotype X, Isolated from Cattle in Nigeria in 2015.

The complete genome sequences of four foot-and-mouth disease viruses of South African territories 1 (SAT 1) serotype are reported. These viruses originate from an outbreak in Nigeria in 2015 and belong to the novel SAT 1 topotype X from the west and central African virus pool.

DIAGNOSIS OF ORF IN WEST AFRICAN DWARF GOATS IN UYO, AKWA IBOM STATE, NIGERIA.

BACKGROUND: Sixty (60) male West African Dwarf goats were reported with clinical signs of enlarged lymph nodes, scabs on the mouth, nose and ears. Two of the goats died and post mortem examination reveals enlarged submandibular lymph nodes and vesicular lesions on the tongue. Clinical diagnosis of Orf has been reported in Nigeria but this report is the confirmatory diagnosis of Orf in a suspected outbreak in an experimental farm in Uyo, Akwa Ibom State, Nigeria using molecular techniques. MATERIALS AND METHODS: Scabs, spleen and lymph node samples from goats suspected to have died from Orf were collected, transported on ice to the laboratory and homogenized. The DNA was extracted using QIAmp DNA minikit (Qiagen) according to the manufacturer's instructions. Orf virus (ORFV) was amplified using published ORFVspecific primers by PCR. RESULTS: Morbidity and mortality were 100% and 3.3% respectively, while ORFV was detected by PCR. Diagnosis of Orf was confirmed based on clinical signs of enlarged lymph nodes, scabs on the mouth, nose and ears, necropsy findings of enlarged submandibular lymph nodes and vesicular lesions on the tongue and PCR results. CONCLUSION: This may be the first report of molecular diagnosis of Orf in Nigeria. The 100% morbidity and 3.3% mortality rate is higher than previously reported thus Orf is becoming of greater economic importance than previously thought. It is therefore recommended that routine laboratory diagnosis of Orf be carried nationwide to determine the prevalence of Orf in Nigeria.

Spatial pattern of foot-and-mouth disease virus serotypes in North Central Nigeria.

AIM: This study aimed to determine the foot-and-mouth disease virus (FMDV) serotypes circulating, the prevalence of FMDV serotypes, and the spatial distribution of FMDV among sedentary and pastoral cattle herds in the North-Central Nigeria. MATERIALS AND METHODS: A cross-sectional study was undertaken, during which a total of 155 sera that tested positive for foot-and-mouth disease (FMD) 3ABC non-structural protein antibodies were selected and screened for FMD structural protein serotypes, A, O, SAT 1, and SAT 2 using a solid-phase competitive enzyme-linked immunosorbent assay (ELISA). Epithelial tissue specimens were collected during outbreak investigations which were tested for FMD using an antigen capture ELISA for serotype A, O, SAT 1, and SAT 2. RESULTS: An overall serotype-specific prevalence of 79.35 (95% confidence interval [CI]: 72.4-85.18) was recorded for serotype O, 65.2% (95% CI: 57.41-72.3) for serotype A, 52.9% (95% CI: 45.03-60.67) for SAT 2, and 33.55% (95% CI: 26.45-41.26) for SAT 1. Evidence of exposure to multiple FMDV serotypes showed that 12.26% of the sera samples had antibodies against four serotypes circulating, 30.97% had antibodies against three serotypes circulating, 22.58% had antibodies against two serotypes, and 17% showed exposure to only one serotype. Clinical specimens (epithelial tissue) collected during outbreak investigations showed that serotype O has the highest proportion of 50% with serotype A - 25%; SAT 2 - 20.8%; and SAT 1 - 4.1%. CONCLUSION: The study detected diffuse and co-circulation of serotypes A, O, SAT 1, and SAT 2 within the study area, and hence the need for the appropriately matched multivalent vaccine is strongly advocated for FMD control in Nigeria.

Status of Laboratory Biosafety and Biosecurity in Veterinary Research Facilities in Nigeria.

BACKGROUND: This study determined current status of laboratory biosafety in Nigerian veterinary research facilities. METHODS: A questionnaire was developed to obtain information from researchers across Nigeria from July 2014 to July 2015. Information regarding demographics, knowledge of laboratory biosafety, availability and proper use of personal protective equipment (PPE), any priority pathogens researched, attitude on and use of standard laboratory practices, and biosafety awareness was obtained using a numeric scoring system. Data were analyzed with descriptive statistics, and univariate and multivariate logistic regression. RESULTS: A total of 74 participants from 19 facilities completed the questionnaire. General knowledge scores ranged from 3 to 28 (out of 28 possible points), with 94.6% of respondents receiving low scores (scores < mean + 1 standard deviation). Very few (17.6%) reported availability or use PPE. Many participants (63.5%) reported no access to biosafety level (BSL)-1-3 facilities. None reported availability of a BSL-4 facility. Knowledge scores pertaining to biosafety management practices ranged from 0 to 14 (out of 14 possible points) with 47.3% of respondents receiving good scores (scores > mean + 1 standard deviation). Only 16.2% of respondents (from four facilities) reported having biosafety officers. Rabies virus was the most researched pathogen (31.1% of respondents). The majority (71.6%) were unaware of laws guiding biosafety. Researchers [odds ratio (OR) = 18.0; 95% confidence interval (CI): 1.63, 198.5; p = 0.023], especially in BSL-2 (OR = 258.5; 95% CI: 12.71, 5256; p < 0.001) facility of research institute (OR = 25.0; 95% CI: 5.18, 120.6; p < 0.001), are more likely to have adequate access to and properly utilize biosafety devices and PPE. CONCLUSIONS: Current knowledge of laboratory biosafety is limited except among a few researchers.

Serological evidence of equine influenza virus in horse stables in Kaduna, Nigeria.

Equine influenza virus (EIV) is a major cause of acute respiratory diseases in horses in most parts of the world that results in severe economic losses. Information on the epidemiology of EIV in tropical Africa is scanty. An enzyme-linked immunosorbent assay (ELISA) was used to detect the presence of influenza A virus nucleoprotein (NP) in 284 horse sera in Kaduna State, Northern Nigeria. The ELISA-positive sera were further examined for hemagglutination inhibition (HI) antibodies to two strains each of H3N8 and H7N3 subtypes of influenza A virus. The results showed that antibodies against influenza A virus nucleoprotein were detected in 60.9% (173 of 284) of horses examined by NP-ELISA. Equine H3 and H7 subtypes were detected in 60% (21 of 35) and 20% (7 of 35) of horse sera respectively across the stables. Adequate quarantine of all imported horses, a national equine influenza surveillance plan and an appropriate EIV control program in Nigeria are recommended to safeguard the large horse population.

Geographic distribution of MERS coronavirus among dromedary camels, Africa.

We found serologic evidence for the circulation of Middle East respiratory syndrome coronavirus among dromedary camels in Nigeria, Tunisia, and Ethiopia. Circulation of the virus among dromedaries across broad areas of Africa may indicate that this disease is currently underdiagnosed in humans outside the Arabian Peninsula.