Sequence Diversity of Tp1 and Tp2 Antigens and Population Genetic Analysis of Theileria parva in Unvaccinated Cattle in Zambia's Chongwe and Chisamba Districts.

East Coast Fever (ECF), caused by Theileria parva, is a major constraint to improved livestock keeping in east and central Africa, including Zambia. To understand the dynamics and determine the candidates for immunization in Zambia's Chongwe and Chisamba districts, a combination of Tp1 and Tp2 gene sequencing and microsatellite analysis using nine markers was conducted from which an abundance of Muguga, Kiambu, Serengeti and Katete epitopes in the field samples was obtained. Phylogenetic analysis showed six (Tp1) and three (Tp2) clusters with an absence of geographical origin clustering. The majority of haplotypes were related to Muguga, Kiambu, Serengeti and Katete, and only a few were related to Chitongo. Both antigens showed purifying selection with an absence of positive selection sites. Furthermore, low to moderate genetic differentiation was observed among and within the populations, and when vaccine stocks were compared with field samples, Chongwe samples showed more similarity to Katete and less to Chitongo, while Chisamba samples showed similarity to both Katete and Chitongo and not to Muguga, Kiambu or Serengeti. We conclude that the use of Katete stock for immunization trials in both Chongwe and Chisamba districts might produce desirable protection against ECF.

Molecular characterization and population genetics of Theileria parva in Burundi's unvaccinated cattle: Towards the introduction of East Coast fever vaccine.

Theileria parva (T. parva) is a protozoan parasite that causes East Coast fever (ECF). The disease is endemic in Burundi and is a major constraint to livestock development. In this study, the parasite prevalence in cattle in six regions namely; Northern, Southern, Eastern, Western, Central and North Eastern was estimated. Furthermore, the sequence diversity of p67, Tp1 and Tp2 genes was assessed coupled with the population genetic structure of T. parva using five satellite markers. The prevalence of ECF was 30% (332/1109) on microscopy, 60% (860/1431) on ELISA and 79% (158/200) on p104 gene PCR. Phylogenetic analysis of p67 gene revealed that only allele 1 was present in the field samples. Furthermore, phylogenetic analysis of Tp1 and Tp2 showed that the majority of samples clustered with Muguga, Kiambu and Serengeti and shared similar epitopes. On the other hand, genetic analysis revealed that field samples shared only two alleles with Muguga Cocktail. The populations from the different regions indicated low genetic differentiation (FST = 0.047) coupled with linkage disequilibrium and non-panmixia. A low to moderate genetic differentiation (FST = 0.065) was also observed between samples and Muguga cocktail. In conclusion, the data presented revealed the presence of a parasite population that shared similar epitopes with Muguga Cocktail and was moderately genetically differentiated from it. Thus, use of Muguga Cocktail vaccine in Burundi is likely to confer protection against T. parva in field challenge trials.

Sequence diversity of cytotoxic T cell antigens and satellite marker analysis of Theileria parva informs the immunization against East Coast fever in Rwanda.

BACKGROUND: East Coast fever (ECF) caused by Theileria parva is endemic in Rwanda. In this study, the antigenic and genetic diversity of T. parva coupled with immunization and field challenge were undertaken to provide evidence for the introduction of ECF immunization in Rwanda. METHODS: Blood collected from cattle in the field was screened for T. parva using ELISA and PCR targeting the p104 gene. Tp1 and Tp2 gene sequences were generated from field samples and from Gikongoro and Nyakizu isolates. Furthermore, multilocus genotype data was generated using 5 satellite markers and an immunization challenge trial under field conditions using Muguga cocktail vaccine undertaken. RESULTS: Out of 120 samples, 44 and 20 were positive on ELISA and PCR, respectively. Antigenic diversity of the Tp1 and Tp2 gene sequences revealed an abundance of Muguga, Kiambu and Serengeti epitopes in the samples. A further three clusters were observed on both Tp1 and Tp2 phylogenetic trees; two clusters comprising of field samples and vaccine isolates and the third cluster comprising exclusively of Rwanda samples. Both antigens exhibited purifying selection with no positive selection sites. In addition, satellite marker analysis revealed that field samples possessed both shared alleles with Muguga cocktail on all loci and also a higher proportion of unique alleles. The Muguga cocktail (Muguga, Kiambu and Serengeti) genotype compared to other vaccine isolates, was the most represented in the field samples. Further low genetic sub-structuring (FST = 0.037) coupled with linkage disequilibrium between Muguga cocktail and the field samples was observed. Using the above data to guide a field immunization challenge trial comprising 41 immunized and 40 control animals resulted in 85% seroconversion in the immunized animals and an efficacy of vaccination of 81.7%, implying high protection against ECF. CONCLUSIONS: Antigenic and genetic diversity analysis of T. parva facilitated the use of Muguga cocktail vaccine in field conditions. A protection level of 81.7% was achieved, demonstrating the importance of combining molecular tools with field trials to establish the suitability of implementation of immunization campaigns. Based on the information in this study, Muguga cocktail immunization in Rwanda has a potential to produce desirable results.

Application of discrete choice experiment to analyze the demand for village chicken paid vaccination service against Newcastle disease in DR Congo.

Newcastle disease (ND) is a highly contagious viral disease that can affect a large number of avian species and cause severe economic loss in many countries. This disease is a major constraint for rural chicken production in most developing countries. In this country, ND is known since the 1940s. It exists in enzootic form in almost all provinces of the country. No systematic long-term control measures have been taken against this devastating disease in village poultry. A discrete choice experiment was carried out to analyze the demand for paid vaccination services, by first identifying the preferences of 320 village chicken keepers from eight sites, four of which had benefitted from such a paid chicken vaccination program against ND and four of which did not. The preference was for a vaccination service carried out following imposed calendar. The public veterinarian was the most preferred professional to ensure the paid vaccination of village chickens. The results led to the design of a profile of paid vaccination service tailored to chicken keepers' expectations. The public veterinarian would supervise vaccination activities, which would be implemented by trained community-based health workers, through collective campaigns at fixed periods in the year. The acceptable price (equivalent to 0.10 US dollar per dose) would allow the service to be set in a sustainable way and might be increased if trust in the delivered service may be built further.

Participatory assessment of paid vaccination campaigns for village chickens against Newcastle disease in Kongo Central province.

In the Democratic Republic of Congo (DRC), where state-driven animal vaccination campaigns are organized only in response to epidemics, the organization of a permanent animal vaccination service is urgently needed. A non-governmental organization has set up an experimental paid vaccination service for village chickens against Newcastle Disease (ND) in the Kongo Central province. This mixed-method study presents a participatory assessment of this experiment, identifying socio-economic factors that influence the decision of chicken keepers to adopt vaccination. The study was conducted in four territories of the province. Qualitative and quantitative data were collected through 12 focus group discussions (FGDs) with professionals of animal health and chicken keepers and 160 semi-structured interviews with chicken keepers, sampled by snowball technique. This participatory process has resulted in the design of a grid for assessing animal vaccination service's performance. Here translating the narratives into a preliminary structured assessment, this grid is an output of the study, to be mobilized for future rapid assessments of the vaccination service in a quantitative prospect. The grid consisted of nine criteria, further composed by 16 items, translated into questions to be asked to chicken keepers and vaccinators. In our study area, fieldworkers enumerated four animal vaccination campaigns during a period of 21 years (except those subject to the present assessment). Around 13% of chicken keepers of our sample had participated in ND vaccination programs. Almost 96% of interviewed chicken keepers expressed their willingness to pay for ND vaccination, and 87% of chicken keepers that vaccinated their chickens perceived the vaccine as effective. Vaccinators estimated that 56% of the chicken keepers who were contacted had actually paid for the vaccination of their chickens. The assessment grid highlighted four points in favor of the sustainability of this service, i.e. the general interest of chickens keepers, vaccine efficacy, vaccine availability and ease of use of the vaccine. Two weak points were identified, viz. the poor access of chicken keepers to information and the weak motivation of vaccinators. The vaccine coverage was calculated within the sample at 13.1%. Paid vaccination campaign for village chicken in Kongo Central obtained a performance score of 62.8%, with the highest score in Kwilu-Ngongo (73.1%) and the lowest in Kasangulu (52.4%). Two factors of adoption of vaccination were identified as statistically significant, i.e. chicken housing and territory. Significant differences appeared between territories in access to information for chicken keepers and in vaccinators motivation. The priorities for the improvement of this service appear to be awareness raising among chicken keepers and increasing vaccinators' motivation.

Determinants of dog owner-charged rabies vaccination in Kinshasa, Democratic Republic of Congo.

Rabies is a preventable fatal disease that causes about 61,000 human deaths annually around the world, mostly in developing countries. In Africa, several studies have shown that vaccination of pets is effective in controlling the disease. An annual vaccination coverage of 70% is recommended by the World Health Organization as a control threshold. The effective control of rabies requires vaccination coverage of owned dogs. Identification of the factors determining dog owners' choice to vaccinate is necessary for evidence-based policy-making. However, for the Democratic Republic of Congo (DRC), the limited data on rabies vaccination coverage makes it difficult for its control and formulation of appropriate policies. A cross-sectional study was conducted in Kinshasa (Lemba commune) with dog-owning households and owned dogs as study populations. The association between dog vaccination and independent factors (household socio-demographics characteristics, dog characteristics, knowledge of rabies and location of veterinary offices/clinics) was performed with Epi-info 7. The Odds Ratio (OR) and p-value < 0.05 were used to determine levels of significance. A total of 166 households owning dogs and 218 owned dogs were investigated. 47% of the dogs had been vaccinated within one year preceding the survey which is higher than the critical coverage (25 to 40%) necessary to interrupt rabies transmission but below the 70% threshold recommended by WHO for control. The determinants of vaccination included socio-economic level of the household (OR = 2.9, p<0.05), formal education level of the dog owner (OR = 4, p<0.05), type of residence (OR = 4.6, p<0.05), knowledge of rabies disease (OR = 8.0, p<0.05), knowledge of location of veterinary offices/clinics (OR = 3.4, p<0.05), dog gender (OR = 1.6, p<0.05) and dog breed (OR = 2.1, p<0.05). This study shows that the vaccination coverage in this area can easily reach the WHO threshold if supplemented by mass vaccination campaigns.

Safety and immunogenicity of Onderstepoort Biological Products' Rift Valley fever Clone 13 vaccine in sheep and goats under field conditions in Senegal.

This blinded field safety study was conducted in Senegal to assess safety and immunogenicity of administration of the registered dose of Rift Valley fever virus (RVFV) Clone 13 vaccine (Onderstepoort Biological Products) to sheep and goats of West African breeds under natural conditions. A total of 267 small ruminants (220 sheep, 47 goats) were included; half received RVFV Clone 13 vaccine at the recommended dose and half received the diluent (as placebo) only. The study was performed on three commercial farms in the northern and eastern region of Senegal in accordance with veterinary good clinical practices. The animals were observed daily for 3 days after vaccination, and then weekly for 1 year. In both sheep and goats vaccinated against RVFV seroconversion rates above 70% were recorded. No seroconversion related to RVFV was observed in placebo-treated animals. No statistically significant differences were determined between placebo and vaccinated groups for mean rectal temperatures for the first 3 days after administration (p > 0.05). No abnormal clinical signs related to treatment were noted, and only one slight injection site reaction was observed in one vaccinated animal for 2 days after vaccination. Out of 176 births assessed over 1 year (93 from the vaccinated group, 83 from the placebo group), 9 were abnormal in the placebo group and 3 in the vaccinated group (p > 0.05). The frequency of adverse events was similar in the placebo and vaccinated groups. RVFV Clone 13 vaccine administered according to the manufacturer's instructions was safe and well tolerated in West African breeds of sheep and goats, including animals of approximately 6 months of age and pregnant females, under field conditions in Senegal. Antibody levels persisted up to 1 year after vaccination.