Epidemiology of Foot and Mouth Disease in Ethiopia: a Retrospective Analysis of District Level Outbreaks, 2007-2012.

This study aimed at determining the incidence, distribution, risk factors, and causal serotypes of foot and mouth disease (FMD) outbreaks in Ethiopia based on 5 years of retrospective outbreak data (September 2007 until August 2012). District level outbreak data were collected from 115 randomly selected districts using a questionnaire administered to district animal health officers. The national incidence of FMD outbreaks during the study period was 1.45 outbreaks per five district years. Outbreaks were geographically widespread affecting all major regional states in the country and were more frequent in the central, southern, and southeastern parts of the country. Neither long-term nor seasonal trends were observed in the incidence of outbreaks. A mixed effects logistic regression analysis revealed that the type of production system (market oriented system versus subsistence systems), presence of a major livestock market and/or route, and adjacency to a national parks or wildlife sanctuary were found to be associated with increased risk of outbreaks in the districts. FMD virus serotypes O, A, SAT 2, and SAT 1 were identified as the causal serotypes of the outbreaks during the study period. Whereas O was the dominant serotype, SAT 2 was the serotype that showed increase in relative frequency of occurrence. The estimated incidence of outbreaks is useful in assessing the economic impacts of the disease, and the identified risk factors provide important knowledge to target a progressive FMD control policy for Ethiopia.

Introduction of the AmpliChip CYP450 Test to a South African cohort: a platform comparative prospective cohort study.

BACKGROUND: Adverse drug reactions and lack of therapeutic efficacy associated with currently prescribed pharmacotherapeutics may be attributed, in part, to inter-individual variability in drug metabolism. Studies on the pharmacogenetics of Cytochrome P450 (CYP) enzymes offer insight into this variability. The objective of this study was to compare the AmpliChip CYP450 Test® (AmpliChip) to alternative genotyping platforms for phenotype prediction of CYP2C19 and CYP2D6 in a representative cohort of the South African population. METHODS: AmpliChip was used to screen for thirty-three CYP2D6 and three CYP2C19 alleles in two different cohorts. As a comparison cohort 2 was then genotyped using a CYP2D6 specific long range PCR with sequencing (CYP2D6 XL-PCR + Sequencing) platform and a PCR-RFLP platform for seven CYP2C19 alleles. RESULTS: Even though there was a low success rate for the AmpliChip, allele frequencies for both CYP2D6 and CYP2C19 were very similar between the two different cohorts. The CYP2D6 XL-PCR + Sequencing platform detected CYP2D6*5 more reliably and could correctly distinguish between CYP2D6*2 and *41 in the Black African individuals. Alleles not covered by the AmpliChip were identified and four novel CYP2D6 alleles were also detected. CYP2C19 PCR-RFLP identified CYP2C19*9,*15, *17 and *27 in the Black African individuals, with *2, *17 and *27 being relatively frequent in the cohort. Eliminating mismatches and identifying additional alleles will contribute to improving phenotype prediction for both enzymes. Phenotype prediction differed between platforms for both genes. CONCLUSION: Comprehensive genotyping of CYP2D6 and CYP2C19 with the platforms used in this study, would be more appropriate than AmpliChip for phenotypic prediction in the South African population. Pharmacogenetically important novel alleles may remain undiscovered when using assays that are designed according to Caucasian specific variation, unless alternate strategies are utilised.

Equine encephalosis virus: evidence for circulation beyond southern Africa.

Prior to the recent outbreak of equine encephalosis in Israel in 2009, equine encephalosis virus (EEV) had only been isolated from equids in South Africa. In this study we show the first evidence for the circulation of EEV beyond South Africa in Ethiopia, Ghana and The Gambia, indicating that EEV is likely to be freely circulating and endemic in East and West Africa. Sequence analysis revealed that the EEV isolate circulating in The Gambia was closely related to an EEV isolate that was isolated from a horse from Israel during the EEV outbreak in 2009, indicating that the two viruses have a common ancestry. Interestingly horses in Morocco tested negative for EEV antibodies indicating that the Sahara desert may be acting as a geographical barrier to the spread to the virus to North African countries. This evidence for EEV circulation in countries in East and West Africa sheds light on how the virus may have reached Israel to cause the recent outbreak in 2009.

Foot and mouth disease in the Borana pastoral system, southern Ethiopia and implications for livelihoods and international trade.

Participatory epidemiology (PE) was used on the Borana plateau of southern Ethiopia to understand pastoralist's perceptions of the clinical and epidemiological features of foot and mouth disease (FMD) in cattle. Matrix scoring showed good agreement between informant groups on the clinical signs of acute and chronic FMD, and findings were cross-checked by clinical examination of cattle and assessment of previous clinical FMD at herd level by detection of antibody to non structural proteins of FMD virus. The positive predictive value of pastoralist's diagnosis of FMD at herd level was 93.1%. The annual age-specific incidence and mortality of acute FMD in 50 herds was estimated using proportional piling. The estimated mean incidence of acute FMD varied from in 18.5% in cattle less than two years of age to 14.0% in cattle three to four years of age. The estimated mean mortality due to acute FMD varied from 2.8% in cattle less than two years of age to 0.3% in cattle three of age or older. Pearson correlation coefficients for acute FMD by age group were -0.12 (p>0.05) for incidence and -0.59 (p<0.001) for mortality. Estimates of the annual incidence of chronic FMD varied from 0.2% in cattle less than two years of age to 1.8% in cattle three to four years of age. The Pearson correlation coefficient for the incidence of chronic FMD by age group was 0.47 (p<0.001). Outbreaks of FMD peaked in Borana cattle during the two dry seasons and were attributed to increased cattle movement to dry season grazing areas. The mean seroprevalence of FMD was estimated at 21% (n=920) and 55.2% of herds (n=116) tested seropositive. Serotyping of 120 seropositive samples indicated serotypes O (99.2%), A (95.8%), SAT 2 (80%) and C (67.5%). The endemic nature of FMD in Borana pastoral herds is discussed in terms of the direct household-level impact of the disease, and the increasing export of cattle and chilled beef from Ethiopia.

Comparison of SAT-1 foot-and-mouth disease virus isolates obtained from East Africa between 1971 and 2000 with viruses from the rest of sub-Saharan Africa.

Foot-and-mouth disease serotype SAT-1 seems to be endemic in many sub-Saharan African countries. Phylogenetic analysis using the 1D gene of 51 SAT-1 isolates from East, West and southern Africa indicated the presence of at least 6 lineages and 11 genotypes with linkages between various geographical regions of the subcontinent. Differences were observed between countries in East Africa, the main focus of this study, with individual countries suffering outbreaks from isolates belonging to various genotypes, which is evidence of reintroduction of strains and long-term circulation of outbreak viruses. The amount of variation observed has significant implications for disease control on the subcontinent.

Study of the genetic heterogeneity of SAT-2 foot-and-mouth disease virus in sub-Saharan Africa with specific focus on East Africa.

The epidemiology of serotype SAT-2 foot-and-mouth disease was investigated in sub-Saharan Africa by phylogenetic analysis using the 1D gene encoding the major antigenic determinant. Fourteen genotypes were identified of which three are novel and belong to East Africa, bringing the total number of genotypes for that region to eight. The genotypes clustered into three lineages that demonstrated surprising links between East, southern and south-western Africa. One lineage was unique to West Africa. These results established numerous incursions across country borders in East Africa and long term conservation of sequences for periods up to 41 years. Ethiopia, Kenya and Uganda have all experienced outbreaks from more than one unrelated strain, demonstrating the potential for new introductions. The amount of variation observed within this serotype nearly equalled that which was found between serotypes; this has severe implications for disease control using vaccination.

Molecular epidemiology of serotype O foot-and-mouth disease virus isolated from cattle in Ethiopia between 1979-2001.

Partial 1D gene characterization was used to study phylogenetic relationships between 17 serotype O foot-and-mouth disease (FMD) viruses in Ethiopia as well as with other O-type isolates from Eritrea, Kenya, South and West Africa, the Middle East, Asia and South America. A homologous region of 495 bp corresponding to the C-terminus end of the 1D gene was used for phylogenetic analysis. This study described three lineages, viz. African/Middle East-Asia, Cathay and South American. Within lineage I, three topotypes were defined, viz. East and West Africa and the Middle East-Asia together with the South African isolate. The Ethiopian isolates clustered as part of topotype I, the East African topotype. Two clades (based on < 12 % nucleotide difference) A and B were identified within the East African isolates, with clade A being further classified into three significant branches, A1 (80% bootstrap support), A2 (89% bootstrap support) and A3 (94% bootstrap support). Clade B consisted of two Kenyan isolates. Within topotype I, the 17 Ethiopian isolates showed genetic heterogeneity between themselves with sequence differences ranging from 4.6-14 %. Lineage 2 and 3 could be equated to two significant topotypes, viz. Cathay and South America. Comparison of amino acid variability at the immunodominant sites between the vaccine strain (ETH/19/77) and other Ethiopian outbreak isolates revealed variations within these sites. These results encourage further work towards the reassessment of the type O vaccine strain currently being used in Ethiopia to provide protection against field variants of the virus.

Multiplication of the V4 strain of Newcastle disease virus in Madin Derby bovine kidney cells.

This study describes a reproducible cell culture system that permits the growth and secondary multiplication of the V4 strain of Newcastle disease virus. Allantoic fluid, magnesium chloride and diethylaminoethyl dextran were incorporated in Dulbecco's modified Eagle's medium to encourage secondary viral multiplication without adversely affecting healthy Madin Derby bovine kidney cell growth.

Development of solid phase antigen for indirect ELISA for the detection of specific antibody responses to infection with Newcastle disease virus.

A simple and inexpensive method of antigen preparation by ultrafiltration was investigated using the V4 strain of Newcastle disease virus. The antigen designated XM300 was used in an indirect enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to Newcastle disease virus in chicken serum. The assay was evaluated using both experimental and field sera, as well as reference control reactor and non-reactor sera. Antigen prepared by the ultrafiltration method was compared with antigen prepared by ultracentrifugation and the ultrafiltration antigen was found to react specifically with Newcastle disease virus antiserum in this ELISA system. This antigen preparation technique is also suitable for use in developing countries. The ELISA provides an excellent method for measuring antibodies in the early stages of infection in serum samples from experimentally infected chickens. More than 14.58 % of the total serum samples which failed to be recognized as reactors by the conventional haemagglutination inhibition test were detected in the ELISA.

Parasitological prevalence of bovine trypanosomosis in Kindo Koisha district, Wollaita zone, south Ethiopia.

A cross sectional survey to determine the distribution and prevalence of trypanosomosis was conducted in Kindo Koisha district, in the Wollaita zone in southern Ethiopia. A total of 1 008 adult cattle was examined at eight different localities. Dark field examination of the buffy coat, as well as stained thin blood film examination and packed cell volume (PCV) evaluation were the diagnostic techniques used. The overall prevalence of bovine trypanosomosis was 15 %. Among the positive animals, 108 (71.1%), 43 (28.4%) and 1 (0.6%) were due to Trypanosoma vivax, Trypanosoma congolense and mixed infection (T. vivax and T. congolense), respectively. The infection rate of T. vivax and T. congolense varied significantly (P < 0.01). The mean PCV of the positive and negative animals ranged between 18.3-32.1% and 26.8-33.4%, respectively. The mean PCV of negative animals (28 %) was significantly higher than the mean PCV of positive animals (22.3%) (P < 0.001). There was an inverse association of PCV with the prevalence of trypanosomosis (P > 0.05). The herd average PCV values of each site decreased with increasing proportion of the positive herds of that particular site. Of the diagnostic tests employed, the microhaematocrit buffy coat technique is relatively sensitive and it has an added advantage of indicating the general condition of the animal by haematocrit measurement. In view of the risk of trypanosomosis, a control intervention through the strategic application of appropriate trypanocidal drugs is recommended. A tsetse fly control scheme to reduce host-tsetse fly contact is equally as important as chemotherapy and chemoprophylaxis against trypanosomosis.

Carob pod (Ceratonia siliqua) meal in geese diets.

1. The apparent and true metabolisable energy values of carob pods meal for geese were measured to be 6.1 MJ/kg and 6.6 MJ/kg respectively. 2. Performance from 5 to 12 weeks was examined in geese fed on four diets containing 0, 100, 200 and 300 g/kg of carob pods meal. 3. The inclusion of carob pods meal up to 200 g/kg in geese diets did not affect the performance. 4. At 300 g/kg performance was highly depressed. 5. The digestibility of protein in the diets decreased linearly with an increase in the level of inclusion of carob pods meal. 6. The length of small intestine, large intestine and caeca and the weight of gizzard expressed per kg of body weight increased with an increase in the level of carob pods meal, which is rich in fibre, in the diets.