First report of co-infections of Marek's disease virus and chicken infectious anaemia virus in poultry flocks in Nigeria.

Marek's disease (MD) and chicken infectious anaemia (CIA) are viral immunosuppressive diseases of poultry caused by the MD virus (MDV) and CIA virus (CIAV) respectively. Despite vaccination against MD, the incidence of the disease in vaccinated poultry flocks in Nigeria persists. However, underlying factors like co-infection with CIAV have not been investigated in the country. This study was designed to investigate possible co-infections of MDV and CIAV in poultry flocks in Nigeria. In 2016, tumorous tissue samples were collected from suspected cases of MD at necropsy in Jos, Plateau State, Nigeria. The samples collected were fixed in formalin for histopathological examination, genomic DNA was extracted from a second part and analysed by polymerase chain reaction (PCR), targeting the meq and VP1 genes of the MDV and CIAV, respectively. The histology results revealed that the cutaneous and proventricular lymphomas were characterized by large numbers of mononuclear cellular infiltrates admixed with heterophils. The PCR results revealed that MDV was detected in 66.7% (16/24), CIAV in 45.8% (11/24), and co-infections of MDV and CIAV were detected in 45.8% (11/24) of the samples analysed. In addition, co-infections of MD and CIA were recorded in 100% (6/6) and 27.7% (5/18) of broilers and layer/pullet' samples respectively. Phylogenetic analysis of the meq gene sequences revealed that the Nigerian MDV clusters with very virulent MDV from Egypt and Italy. While, CIAV sequences were genotype II and genotype III and clustered with CIAVs from Cameroon and China. This is the first report of co-infections of MD and CIA in Nigeria.

Characterization of a Novel African Swine Fever Virus p72 Genotype II from Nigeria.

African swine fever (ASF) is a high-consequence transboundary hemorrhagic fever of swine. It continues to spread across the globe causing socio-economic issues and threatening food security and biodiversity. In 2020, Nigeria reported a major ASF outbreak, killing close to half a million pigs. Based on the partial sequences of the genes B646L (p72) and E183L (p54), the virus responsible for the outbreak was identified as an African swine fever virus (ASFV) p72 genotype II. Here, we report further characterization of ASFV RV502, one of the isolates obtained during the outbreak. The whole genome sequence of this virus revealed a deletion of 6535 bp between the nucleotide positions 11,760-18,295 of the genome, and an apparent reverse complement duplication of the 5' end of the genome at the 3' end. Phylogenetically, ASFV RV502 clustered together with ASFV MAL/19/Karonga and ASFV Tanzania/Rukwa/2017/1 suggesting that the virus responsible for the 2020 outbreak in Nigeria has a South-eastern African origin.

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.

Live pig markets are hotspots for spread of African swine fever virus in Nigeria.

Livestock trading through live animal markets are potential pathways for the introduction and spread of economically important pathogens like the African swine fever virus (ASFV) to new areas in several countries. Due to the high demand for live pigs in Nigeria both for restocking and slaughter, live pigs are sold at designated live pig markets (LPM) in the country. This involves movement of pigs over long distances. Despite, reports of ASF outbreaks following restocking of pigs bought from LPMs, there is paucity of information on the role of LPMs in the epidemiology of ASF. In this study, data and pig samples (whole blood, sera, tissue) were collected from 4 selected LPMs in Nigeria (Dawaki, Katsit, Numan and Pandam) between 2019 and 2020. Samples were analysed by polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Four genes of ASFV positive samples were characterized to identify the circulating genotypes. Results revealed trade activities involving transportation of pigs from these selected markets to 42 major cities and towns in thirteen (13) States of Nigeria. PCR results revealed an overall ASF prevalence of 10.77% (66/613). ASFV was confirmed by PCR in all the selected LPMs with a prevalence rate of 3.13%-23.81%. The phylogeny revealed genotype I and serogroup 4 based on the p72 protein that encodes the B646L gene and the EP402R gene encoding the CD2V. While sequence analysis of CVR of B602L gene revealed 8 tetrameric repeats variants, six of which have never been reported in Nigeria. Analysis of sera samples recorded a seroprevalence of 6.9% (16/217) within the study period. Findings from this study show that LPM are hotspots and channels for transmission and continuous spread of ASFV in Nigeria. Therefore, for ASF to be controlled in Nigeria, disease surveillance and regulation at LPMs are critical.

Financial impact of sheeppox and goatpox and estimated profitability of vaccination for subsistence farmers in selected northern states of Nigeria.

Sheeppox and goatpox (SGP) are important transboundary diseases, endemic in Nigeria, causing severe clinical manifestations, impacting production, and resulting in economic losses. Vaccination is an effective control measure against SGP in endemic countries but is not currently implemented in Nigeria. This study aimed to estimate SGP financial impact and assess economic viability of SGP vaccination at the herd and regional level under different scenarios in Northern Nigeria. Integrated stochastic production and economic herd models were developed for transhumance and sedentary herds. Models were run for two disease scenarios (severely and slightly affected) and with and without vaccination, with data parameterisation from literature estimates, field survey and authors' experience. Herd-level net financial impact of the disease and its vaccination was assessed using gross margin (GM) and partial budget analyses. These were then used to assess regional financial impact of disease and profitability of a 3-year vaccination programme using a cost-benefit analysis. The regional-analysis was performed under 0 %, 50 % and 100 % government subsidy scenarios; as a standalone programme or in combination with other existing vaccination programmes; and for risk-based and non-risk-based intervention. Median SGP losses per reproductive female were £27 (90 % CI: £31-£22), and £5 (90 % CI: £7-£3), in sedentary, and £30 (90 % CI: £41-21), and £7 (90 % CI: £10-£3), in transhumance herds, for severely and slightly affected scenarios respectively. Selling animals at a reduced price, selling fewer young animals, and reduced value of affected animals remaining in the herd were the greatest contributors to farmer's SGP costs. SGP-affected herds realised a GM reduction of up to 121 % in sedentary and 138 % in transhumance. Median estimated regional SGP cost exceeded £24 million. Herd-level median benefits of vaccination per reproductive female were £23.76 (90 % CI: £19.28-£28.61), and £4.01 (90 % CI: £2.36-£6.31), in sedentary, and £26.85 (90 % CI: £17.99-£37.02) and £7.45 (90 % CI: £3.47-£15.14) in transhumance herds, in severely and slightly affected scenarios, respectively. Median benefit: cost ratio (BCR) for severely affected herds at 50% subsidies was 6.62 (90% CI: 5.30-8.90) for sedentary, and 5.14 (90% CI: 3.31-13.81) for transhumance herds. The regional SGP vaccination standalone programme BCR: 7-27, regional SGP vaccination with existing vaccination programme BCR: 7-228 and vaccinating high-risk areas BCR: 19-439 were found to be economically viable for all subsidy levels explored. Vaccinating low-risk areas only realised benefits with 100 % of government subsidies. This study further increases understanding of SGP's impact within Northern Nigeria and demonstrates vaccination is an economically viable control strategy at the herd-level and also regionally, depending on the strategy and government subsidy levels considered.

Household and animal factors associated with sheeppox and goatpox sero-prevalence and identification of high-risk areas in selected States of northern Nigeria.

Sheeppox and goatpox (SGP) are transboundary, highly contagious diseases affecting sheep and goats with characteristic clinical signs. SGP affect populations of small ruminants in Africa, Asia and the Middle East and, as a result, threaten farmers' livelihoods. Despite their importance, studies looking at factors that increase the risk of sheeppox-virus (SPPV) and goatpox-virus (GTPV) exposure and infection are limited. A cross-sectional study was conducted in three states of Northern Nigeria (Bauchi, Kaduna and Plateau) to determine the sero-prevalence and spatial patterns of SGP, and identify risk factors for SPPV/GTPV exposure at animal and household level. Sera samples were collected from 1,800 small ruminants from 300 households. Data on putative risk factors were collected using a standardised questionnaire. Twenty-nine small ruminants were sero-positive to SGP - apparent weighted sero-prevalence 2.0 %; 95 % C.I. 1.1-.3.0 %. Sero-positive animals came from 19 (6.3 %) households. Analysis of the questionnaire showed that a fifth (20.3 %) of farmers claimed to have experienced SGP outbreaks previously in their flocks, with 33 (1.8 %) of the individual animals sampled in this study reported to have had clinical signs. At animal level, the odds of being sero-positive were higher in older animals (>24months; OR = 8.0, p = 0.008 vs ≤24 months) and small ruminants with a history of clinical SGP (OR = 16.9, p = 0.01). Bringing new small ruminants into the household and having a history of SGP in the flock were the main factors identified at household level. Households were less likely to be sero-positive if the time between bringing animals into the household and sampling was over a year (PR = 0.31, p = 0.05), while households with a history of SGP were more likely to be sero-positive regardless of the timeframe. Important spatial heterogeneity was found. The Bayes smooth rate ranged from 0.06 to 4.10 % across local government areas (LGA), with LGA in the north-east or north-west of the study area identified as hot-spots for SGP exposure. Results from this study shed new light on the understanding of SGP epidemiology and provide key inputs to design risk-based surveillance and intervention programmes in the area.

First-Time Presence of African Swine Fever Virus Genotype II in Nigeria.

A confirmed African swine fever (ASF) outbreak in Nigeria was further investigated by partial sequencing of the B464L and E183L genes of ASF virus (ASFV). Results revealed the first-time presence of ASFV genotype II in Nigeria and West Africa. This finding has serious implications for control measures and food security.

Laboratory diagnosis of a new outbreak of acute African swine fever in smallholder pig farms in Jos, Nigeria.

African swine fever (ASF) is a highly contagious fatal infectious disease of pigs and wild suids. The disease has a worldwide occurrence and significant impact on pig production. Two adult intensively raised large white boars from two farms in Jos with a history of sudden death were diagnosed of ASF between July and August 2019. Post-mortem examination of carcasses grossly showed splenomegaly, haemorrhagic lymphadenitis and hepatomegaly with severe congestion. The kidneys were enlarged and had generalized petechiae and blood clot in the pelvis. The heart was moderately enlarged. Microscopic examination of the spleen and lymph nodes revealed severe lymphocytic depletion, haemorrhage and severe haemosiderosis. The liver was severely congested with focal coagulative necrosis of the hepatocytes. The kidneys were severely congested and showed renal tubular necrosis with few tubular protein casts. Tissue samples were confirmed to be positive for African swine fever virus (ASFV) by polymerase chain reaction (PCR) assay, and phylogenetic analysis revealed that the isolate belonged to genotype I.

Avian leukosis virus subgroup - J as a contaminant in live commercially available poultry vaccines distributed in Nigeria.

Globally, vaccines are used to prevent and control the menace of infectious diseases in livestock with some reported to be inadvertently contaminated with extraneous agents (EAs). With the aim of screening and characterizing for some selected EAs, 44 live viral poultry vaccines were randomly selected based on availability. The vaccines comprised 14 manufacturers in 10 different countries including Nigeria were screened by Polymerase Chain Reaction. In 9% (4/44) of the vaccines, contamination with only avian leukosis virus (ALV) subgroup J (ALV-J) was recorded. Other exogenous ALV subgroups, chicken infectious anemia and infectious laryngotracheitis viruses were absent. The EAs was found in infectious bursal disease (n = 1), Fowlpox (n = 2) and Mareks disease (n = 1) vaccines. Phylogenetic analysis of the ALV-J env gene showed clustering with contemporary group I and II. The result underscores the importance of screening vaccines to avoid the introduction and spread of EAs that could pose a threat to poultry production.