Molecular characterization of Newcastle disease virus vaccines in Nigeria.

Background and Aim: Newcastle disease (ND) caused by ND virus (NDV) is a serious impediment to effective poultry production in developing countries such as Nigeria. Despite employing vaccination and other control measures to curtail this disease, its severe forms still persist. This study aimed to confirm the virus strains in the NDV vaccine brands commonly used in Nigeria. Materials and Methods: We employed reverse-transcription polymerase chain reaction (RT-PCR), sequencing, and sequence analysis to characterize NDV strains in four NDV vaccines commonly used in Nigeria. Fragments of 300 bp from NDV fusion genes from the vaccines were amplified. Polymerase chain reaction products were sequenced and analyzed using multiple sequence alignment and phylogenetic analyses to characterize the vaccine viruses as pathotypes. Results: All the vaccines gave positive results, confirming the presence of NDV. Multiple sequence alignment and phylogenetic analyses revealed that two of the vaccines had the lentogenic pathotype, while the other two had the mesogenic or velogenic pathotype. Conclusion: This study provides information to facilitate strategies for regular control of the quality of vaccines in Nigeria.

External quality assessment of Rift Valley fever diagnosis in countries at risk of the disease: African, Indian Ocean and Middle-East regions.

Rift Valley fever virus (RVFV), an arbovirus belonging to the Phlebovirus genus of the Phenuiviridae family, causes the zoonotic and mosquito-borne RVF. The virus, which primarily affects livestock (ruminants and camels) and humans, is at the origin of recent major outbreaks across the African continent (Mauritania, Libya, Sudan), and in the South-Western Indian Ocean (SWIO) islands (Mayotte). In order to be better prepared for upcoming outbreaks, to predict its introduction in RVFV unscathed countries, and to run efficient surveillance programmes, the priority is harmonising and improving the diagnostic capacity of endemic countries and/or countries considered to be at risk of RVF. A serological inter-laboratory proficiency test (PT) was implemented to assess the capacity of veterinary laboratories to detect antibodies against RVFV. A total of 18 laboratories in 13 countries in the Middle East, North Africa, South Africa, and the Indian Ocean participated in the initiative. Two commercial kits and two in-house serological assays for the detection of RVFV specific IgG antibodies were tested. Sixteen of the 18 participating laboratories (88.9%) used commercial kits, the analytical performance of test sensitivity and specificity based on the seroneutralisation test considered as the reference was 100%. The results obtained by the laboratories which used the in-house assay were correct in only one of the two criteria (either sensitivity or specificity). In conclusion, most of the laboratories performed well in detecting RVFV specific IgG antibodies and can therefore be considered to be prepared. Three laboratories in three countries need to improve their detection capacities. Our study demonstrates the importance of conducting regular proficiency tests to evaluate the level of preparedness of countries and of building a network of competent laboratories in terms of laboratory diagnosis to better face future emerging diseases in emergency conditions.

A novel MLSA allelic profile 'A15' of Mycoplasma mycoides subsp mycoides in Niger.

Mycoplasma mycoides subsp. mycoides (Mmm) is the aetiological agent of contagious bovine pleuropneumonia (CBPP). The aim of the present study was to identify the profiles of the Mmm strains isolated in Niger using the 'Multilocus Sequence Analysis' (MLSA) typing technique based on polymorphism analysis of housekeeping and non-coding genes. The investigation was conducted on samples (n=22) comprising of lung tissues, lymph node and pleural fluids. Following classical PCR, Mmm positive amplicons (n=6) were identified. These positive amplicons were then amplified using eight loci of the PG1 reference strain (LocPG1-0001, Loc-PG1-0103, Loc-PG1-0287, Loc-PG1-0431, Loc-PG1-0489, Loc-PG1-0523, Loc-PG1-0710 and Loc-PG1-0827). Sequencing followed by the determination of the profile of each strain by the combination of the allele numbers revealed three different MLSA profiles namely; A11, E01 and A15. The profiles A11 and E01 were previously identified. The novel profile identified in this study was named profile A15. The difference was detected while comparing sequences of non-coding loci. This novel profile was named 'A15' according to the similarities with African reference strain profile 'A00' at the seven loci level (loc-0103, loc-0287, loc-0431, loc-0489, loc-0523, loc-0710 and loc-0827). For CBPP control measures, identification and molecular characterization of Mmm strains is very important. Thus, the use of MLSA technique is relevant to identify profiles of Mmm circulating in Niger. Other countries where CBPP is still endemic are encouraged to use a MLSA scheme to address this issue and, most importantly, to rapidly trace back the origin of outbreaks, which will help reduce the transmission and spread of the disease. In addition, mapping the profiles of strains circulating in each of the countries of the sub-region is necessary for effective control of CBPP.

Co-circulation of genotypes XIV.2 and XVIII.2 of avian paramyxovirus-1 (Newcastle disease virus) in backyard poultry in Niger.

The causative agent of Newcastle disease (ND) of poultry is the avian paramyxovirus-1, also commonly known as ND virus (NDV). Like in many developing countries, ND is endemic in Niger and has significant economic impact on commercial and backyard poultry production. NDVs were characterized in Niger between 2006 and 2008 and shown to belong to genotypes XIV.1 and XVII. In order to determine the current situation regarding the virus in Niger, tracheas (n = 384) were collected for the detection of NDV from both healthy (n = 335) and sick (n = 49) backyard poultry in 2019. Of these samples, 24 from sick chickens were positive for NDV by conventional RT-PCR. Sequencing of the fusion protein gene and phylogenetic analysis revealed that the viruses belonged to either genotype XIV.2 or XVIII.2. No NDVs of genotype XIV.1 or XVII were identified in the current study highlighting the dynamic nature of NDV circulation in Niger and the region.

Transboundary spread of equine influenza viruses (H3N8) in West and Central Africa: Molecular characterization of identified viruses during outbreaks in Niger and Senegal, in 2019.

Since November 2018, several countries in West and Central Africa have reported mortalities in donkeys and horses. Specifically, more than 66,000 horses and donkeys have succumbed to disease in Burkina Faso, Chad, Cameroon, The Gambia, Ghana, Mali, Niger, Nigeria, and Senegal. Strangles caused by Streptococcus equi subsp equi, African Horse Sickness (AHS) virus, and Equine influenza virus (EIV) were all suspected as potential causative agents. This study reports the identification of EIV in field samples collected in Niger and Senegal. Phylogenetic analysis of the hemagglutinin and neuraminidase genes revealed that the identified viruses belonged to clade 1 of the Florida sublineage and were very similar to viruses identified in Nigeria in 2019. Interestingly, they were also more similar to EIVs from recent outbreaks in South America than to those in Europe and the USA. This is one of the first reports providing detailed description and characterization of EIVs in West and Central Africa region.

Molecular epidemiology of peste des petits ruminants in Niger: An update.

Like many West African countries, outbreaks of peste des petits ruminants (PPR), an economically important disease of goats and sheep, are regularly reported in Niger. The causative virus, peste des petits ruminants virus (PPRV), can be differentiated into four genetically distinct lineages. A publication in 2018 identified three PPRV lineages circulating in the country in 2001 (lineages I and II) and 2013 (lineage IV), respectively. In this present study, more recent samples were collected from goats and sheep in locations throughout Niger between 2011 and 2017. Twelve PPRV-positive samples were characterized by sequencing of a segment of the nucleocapsid protein (N) gene. Phylogenetic analysis of the sequences identified viruses from lineages II and IV only. The analysis also indicated a shared origin of the viruses from Niger with PPRVs from neighbouring countries suggesting transboundary movement.

Disentangling the role of Africa in the global spread of H5 highly pathogenic avian influenza.

The role of Africa in the dynamics of the global spread of a zoonotic and economically-important virus, such as the highly pathogenic avian influenza (HPAI) H5Nx of the Gs/GD lineage, remains unexplored. Here we characterise the spatiotemporal patterns of virus diffusion during three HPAI H5Nx intercontinental epidemic waves and demonstrate that Africa mainly acted as an ecological sink of the HPAI H5Nx viruses. A joint analysis of host dynamics and continuous spatial diffusion indicates that poultry trade as well as wild bird migrations have contributed to the virus spreading into Africa, with West Africa acting as a crucial hotspot for virus introduction and dissemination into the continent. We demonstrate varying paths of avian influenza incursions into Africa as well as virus spread within Africa over time, which reveal that virus expansion is a complex phenomenon, shaped by an intricate interplay between avian host ecology, virus characteristics and environmental variables.