Seroprevalence and factors associated with bovine viral diarrhea virus (BVDV) infection in dairy cattle in three milksheds in Ethiopia.

This work was conducted to estimate the seroprevalence, to identify potential factors that influence seroprevalence of bovine viral diarrhea virus (BVDV), and to investigate the association between BVDV serostatus and occurrence of reproductive disorders in dairy cattle in three milksheds in Ethiopia. A total of 1379 serum samples were obtained from cattle randomly selected from 149 herds from three milksheds representing central, southern, and western Ethiopia. Sera samples were examined for bovine viral diarrhea virus (BVDV) antibodies using commercial competitive enzyme-linked immunosorbent assay (ELISA) kit. Logistic regression analysis was employed to investigate associations between risk factors and the risk of BVDV seroprevalence, and BVDV serostatus and reproductive disorders. Seroreaction to BVDV antigens was detected in 32.6% of the 1379 cattle and 69.8% of the 149 herds sampled. Factors associated with BVDV seroplevalence were age, breed, and herd size (P < 0.05). Adult cattle ≥ 18 months old had 2.1 (95% CI 1.5, 3.1) times the odds of BVDV seroreaction than younger cattle. Holstein-Friesian (HF) local crosses (OR = 2.1, 95% CI 1.3, 3.4) and HFs (OR = 1.3, 95% CI 0.9, 1.9) were more likely to be seropositive than Jersey and the odds of seropositivity in cattle in large herds with 11 or more animals were higher (OR = 1.8, 95% CI 1.3, 2.5) than the odds of BVDV seropositivity in smaller herds. Seroprevalence was not associated with geographical region (P > 0.05). Risk of reproductive disorders was not affected by BVDV serostatus, except for repeat breeding (P > 0.05). The present study demonstrated that BVDV has wide distribution in the country being detected in all the 15 conurbations and 69.8% of herds involved in the study.

Bovine herpesvirus-1 in three major milk sheds of Ethiopia: Serostatus and association with reproductive disorders in dairy cattle.

Bovine herpesvirus-1 (BHV-1) causes infectious bovine rhinotracheitis (IBR), and infectious pustular vulvovaginitis (IPV) in cows and infectious pustular balanopostitis (IPB) in bulls worldwide. Infection of seronegative cattle with BHV-1 leads to abortion, retention of fetal membranes, increased service per conception, metritis and oophoritis. As part of an ongoing study on infectious causes of reproductive disorders in Ethiopia, this investigation aims at assessing the role of BHV-1 in the disorders and the risk factors affecting its seroprevalence. A cross-sectional study was conducted on a total of 1379 randomly selected dairy cattle from 149 herds. These dairy cattle were sampled from milks sheds of central (n = 555), western (n = 195) and southern (n = 629) Ethiopia. Blocking enzyme-linked immunosorbent assay (B-ELISA) was applied to detect antibodies specific to BHV-1. Additionally, a semi-structured questionnaire was administered and farm records were assessed to capture potential risk factors associated with BHV-1 seropositivity. Univariable and multivariable random-effects logistic regression analyses were used to assess potential risk factors associated with BHV-1 serostatus. Model fitness and reliability were assessed using the Hosmer and Lemeshow method and the receiver operating curve (ROC) respectively. An overall herd level BHV-1 seroprevalence of 81.8% (95% confidence interval (CI): 74.7-87.7%) and individual animal level seroprevalence of 41.0% (95% CI: 38.4-43.7%) were found. In a random-effects multivariable logistic regression model, the seroprevalence of BHV-1 exposure was higher in dairy cattle from breeding (Odds ratio [OR] = 1.3; p = 0.036) than in commercial (OR = 0.9; p = 0.137) and small-holder farms. Geographically, the prevalence was higher in western (OR = 1.4; p < 0.001) and southern Ethiopia (OR = 1.2; p < 0.001) than in central regions. BHV-1 seropositive cows had higher (p < 0.05) odds of clinical reproductive disorders including abortion, retained fetal membranes, stillbirth, birth of weak calf and metritis compared to seronegative cows. Thus, it is suggested that BHV-1 should be considered as differential diagnosis among improved dairy cattle herds with reproductive disorders in Ethiopia.

Seroprevalence of Schmallenberg virus in dairy cattle in Ethiopia.

Schmallenberg virus (SBV) is a recently identified member of the genus Orthobunyavirus of the family Bunyaviridae. It is an arbovirus transmitted by different members of Culicoides spp of biting midges. The virus is more recognized for its effect on reproductive disorders in ruminants characterised by abortion, stillbirth and birth of congenitally defective newborns with hydranencephaly-arthrogryposis syndrome. The current study was undertaken with the objectives of exploring the presence of SBV exposure and identification of factors affecting its distribution among dairy cattle in Ethiopia. A cross-sectional study was conducted on 1379 dairy cattle sampled from 149 dairy herds in central, southern and western Ethiopia during September 2011 to May 2012. Serum samples were examined using competitive enzyme linked immunosorbent assay (cELISA). Data on hypothesised risk factors were collected from farm records where available and semi-structured questionnaire-based interview. The apparent seroprevalence of exposure to SBV was 56.6% (95% confidence interval (CI): 53.9-59.3). True prevalence adjusted for sensitivity and specificity of the cELISA kit used was 58.3% (95% CI 55.7-60.9). Among the sampled herds, 82.6% (95% CI: 75.5-88.3) had at least one seropositive animal. Seropositive cattle were found in all of the 15 conurbations studied. Adult dairy cows [odds ratio (OR)=1.6] were more commonly affected than young heifers. Dairy cattle kept in commercial (OR=1.6) and breeding farms (OR=3.5) and Midland agroecology (OR=2.5) showed statistically significant seroconversion than cattle kept under small-holder dairy farms and Highland agroecology respectively (p<0.05). Reproductive disorders including abortion, retention of the fetal membranes, and metritis were associated with serostatus of SBV. In conclusion, the seroprevalence of SBV is high and widely distributed in the studied parts of Ethiopia. This being the first study of its kind on SBV in Ethiopia, further longitudinal studies on isolation of the virus and its impact on reproductive disorders are recommended.

Serological evidence of Bovine herpesvirus-1, Bovine Viral Diarrhea virus and Schmallenberg virus infections in relation to reproductive disorders in dairy cattle in Ethiopia.

Reproductive disorders in dairy cattle have been noted to be common in urban and peri-urban dairy production system in Ethiopia. The available reports on the causes of these disorders, however, are not conclusive. A case-control study was designed to investigate the possible association of major reproductive disorders in dairy cattle with exposure status to bovine herpesvirus-1 (BHV-1), bovine viral diarrhea virus (BVDV) and Schmallenberg virus (SBV). Cows with history of abortion/stillbirth were considered as cases (n=204) while, those cows with no such history were taken as control (n=359). The serological screening tests used for all the three viruses were blocking enzyme linked immunosorbent assays (B-ELISAs). Of the total 563 samples tested 58.4%, 43.8% and 32.9% were positive for SBV, BHV-1 and BVDV, respectively. Significant difference between cases and controls were noted for SBV (p=0.026) and BHV-1 exposures (p<0.001). The difference noted for BVDV serostatus was not significant (p>0.05). The highest proportion (28.9%) of concurrent exposures was noted for BHV-1 and SBV, followed by SBV and BVDV (21.5%) and BHV-1 and BVDV (20.2%). Evidence of exposures to all the three viruses were detected in 14.4% of the animals. However, significant difference between cases (39.7%) and controls (22.9%) among cattle with multiple sero-positivity was noted only for BHV-1 and SBV (p<0.001). Proportion of uterine infection (p=0.002) and fetal membrane retention (p=0.005) increased in BHV-1 seropositive animals, while repeat breeding was common (p=0.034) among BVDV exposed ones. Seropositive animals to any of the three viruses were detected in all sampled areas and the proportion of cattle with BHV-1 and SBV exposure history had a higher risk to at least one type of the reproductive disorders mentioned compared to the corresponding sero-negative groups.

Bovine tuberculosis in Ethiopia: A systematic review and meta-analysis.

Bovine tuberculosis (bTB) is a known endemic disease of cattle in Ethiopia; however, there is lack of a comprehensive information on the status and distribution of the disease in the country. The objectives of this systematic review and meta-analysis were to provide a pooled prevalence estimate of bTB at a national level, assess the level of in-between variance among study reports and illustrate the spatial distribution pattern in the country. Articles published on bTB from January 2000 to December, 2016 in English language were included in the review. Pubmed, CAB direct, AJOL and Web of Science were the databases used in electronic search. A total of 127 articles were retrieved from online sources, of which 56 articles were selected for data extraction based on the specified inclusion criteria. From these selected published articles, 114 animal level data were extracted for quantitative analysis. A pooled prevalence estimate of bovine tuberculosis in Ethiopia was found to be 5.8% (95% CI: 4.5, 7.5). In a multivariable meta-regression analysis, breed and production system explained 40.9% of the explainable proportion of the in-between study variance computed. The prevalence of bovine tuberculosis in Holstein-Friesians, 21.6% (95% CI: 14.7-30.7), was higher than the prevalence in local zebus 4.1 (95% CI: 3.4-4.9). Cattle kept under intensive and semi-intensive production systems had higher prevalence, 16.6% (95% CI: 12.4-21.6), of bTB than those kept in extensive livestock production system, 4.6 (95% CI: 3.4-6.2). Bovine tuberculosis is widely distributed across major livestock producing regions of Ethiopia. However, no valid data could be retrieved from Benishanul-Gumuz, Harari and Dire Dawa. Data obtained on bTB from Somali and Gambella regional states are also few and further studies are suggested in these regions. In conclusion, this review showed that bTB in cattle in Ethiopia is widespread with high prevalence in intensive and semi-intensive management systsems that keep exotic breeds and their crosses in urban and peri-urban areas. Thus, it is suggested that the design and implementation of bTB control strategies in Ethiopia should prioritize these hotspots in order to reduce the impact of the disease on the growing dairy sector.

Infectious bursal disease: outbreak investigation, molecular characterization, and vaccine immunogenicity trial in Ethiopia.

The study was conducted with the objective of isolation and molecular characterization of infectious bursal disease virus (IBDV) circulating in Ethiopia and to assess the immunogenicity of different commercially available live attenuated IBD vaccines and finally to select the appropriate vaccine strain for the existing IBDV. Outbreak samples collected from different poultry farms with IBD infection between 2013 and 2015 were used for the virus isolation and molecular characterization. IBD vaccine immunogenicity test was conducted using four different commercially available live attenuated IBD vaccine strains: namely D78, B2K, LC75, and EXTREM. Day-old Bowman brown chickens purchased from commercial farm in Debre Zeit were used for the experiment. Serum samples were collected at days 14 and 21 and screened for the presence of maternal IBDv antibodies. The screening test result revealed that most of the chickens from vaccinated progeny were positive at the age of day 14 with mean antibody titer of .42, but declined at day 21 to 0.049 below cut-off point (S/P < 0.3). Chickens were divided into five different groups (four vaccinal and one control) and vaccinated at the age of day 21 and boosted after 14 days. Serum samples were collected and all of them were challenged at their 42 days of age with locally isolated very virulent infectious bursal disease virus (vvIBDV). From four of the vaccine strains used for immunogenicity study, the intermediate plus strains (LC75 and EXTREM) found to be superior and efficiently cross protect against the challenge with locally isolated vvIBDV. The development of clinical signs was studied and post-mortem examinations were conducted both on dead and sacrificed birds. From a total of 25 tissue samples processed for virus isolation on chicken fibroblast cell culture, 95% (18/20) of bursa and 80% (4/5) of the spleen samples showed visible cytopathic effect (CPE). The positive samples were tested by PCR and 19 of them had the expected band (643 bp). Further 11 representative samples were sequenced and confirmed that the circulating virus among poultry population in the country is vvIBDV. The study has recommended to produce vaccine using intermediate plus strains to prevent and control currently circulating vvIBDV.

Risk factors for MERS coronavirus infection in dromedary camels in Burkina Faso, Ethiopia, and Morocco, 2015.

Understanding Middle East respiratory syndrome coronavirus (MERS-CoV) transmission in dromedary camels is important, as they consitute a source of zoonotic infection to humans. To identify risk factors for MERS-CoV infection in camels bred in diverse conditions in Burkina Faso, Ethiopia and Morocco, blood samples and nasal swabs were sampled in February-March 2015. A relatively high MERS-CoV RNA rate was detected in Ethiopia (up to 15.7%; 95% confidence interval (CI): 8.2-28.0), followed by Burkina Faso (up to 12.2%; 95% CI: 7-20.4) and Morocco (up to 7.6%; 95% CI: 1.9-26.1). The RNA detection rate was higher in camels bred for milk or meat than in camels for transport (p = 0.01) as well as in younger camels (p = 0.06). High seropositivity rates (up to 100%; 95% CI: 100-100 and 99.4%; 95% CI: 95.4-99.9) were found in Morocco and Ethiopia, followed by Burkina Faso (up to 84.6%; 95% CI: 77.2-89.9). Seropositivity rates were higher in large/medium herds (≥51 camels) than small herds (p = 0.061), in camels raised for meat or milk than for transport (p = 0.01), and in nomadic or sedentary herds than in herds with a mix of these lifestyles (p < 0.005).

Investigation of Marek's disease virus from chickens in central Ethiopia.

Marek's disease (MD) is a lymphoproliferative and neuropathic disease of domestic chickens and less commonly, turkeys and quails, caused by a highly contagious, cell-associated, oncogenic herpesvirus. In Ethiopia, MD is believed to be introduced with importation of exotic and crossbred to improve the poultry production and has been reported to be a potential threat to the poultry sector both in backyard and commercial farming systems. This study was aimed at isolation and molecular analysis of MD virus isolates circulating in chicken population in the central part of Ethiopia where commercial farms are populated. From September 2013 to January 2014, clinical and post-mortem examination were conducted on diseased chickens suspected of MD virus infection. Representative spleen and feather follicle samples were collected following sterile procedure, and infectious virus isolation was performed using primary chicken fibroblast cell culture. Cell culture inoculated with suspension of pathological samples developed characteristic MD virus cytopathic effect of rounding of the cells and small plaques. Further analysis of the virus was conducted by conventional PCR amplifying the ICP4 gene fragment from eleven tissue samples using MD virus specific primers. PCR products were further sequenced and analyzed. Nucleotide sequence similarity search of the local isolates resulted a high degree of sequence similarity with Gallid Herpes virus type 2 strain (Marek's disease virus type 1, JN034558). To our knowledge, the present study is the first report conducted on virus isolation and molecular characterization of MD virus isolates circulated in Ethiopia. Eleven ICP4-like gene fragment (318 bp) sequences generated in the present study were uploaded in the public database (KU842366-76). Further research on virus isolation, genetic characterization, and infection dynamics is recommended targeting chickens of all age groups reared in different agro-ecological zones under different production system.

Genetic characterization of poxviruses in Camelus dromedarius in Ethiopia, 2011-2014.

Camelpox and camel contagious ecthyma are infectious viral diseases of camelids caused by camelpox virus (CMLV) and camel contagious ecthyma virus (CCEV), respectively. Even though, in Ethiopia, pox disease has been creating significant economic losses in camel production, little is known on the responsible pathogens and their genetic diversity. Thus, the present study aimed at isolation, identification and genetic characterization of the causative viruses. Accordingly, clinical case observations, infectious virus isolation, and molecular and phylogenetic analysis of poxviruses infecting camels in three regions and six districts in the country, Afar (Chifra), Oromia (Arero, Miyu and Yabello) and Somali (Gursum and Jijiga) between 2011 and 2014 were undertaken. The full hemagglutinin (HA) and partial A-type inclusion protein (ATIP) genes of CMLV and full major envelope protein (B2L) gene of CCEV of Ethiopian isolates were sequenced, analyzed and compared among each other and to foreign isolates. The viral isolation confirmed the presence of infectious poxviruses. The preliminary screening by PCR showed 27 CMLVs and 20 CCEVs. The sequence analyses showed that the HA and ATIP gene sequences are highly conserved within the local isolates of CMLVs, and formed a single cluster together with isolates from Somalia and Syria. Unlike CMLVs, the B2L gene analysis of Ethiopian CCEV showed few genetic variations. The phylogenetic analysis revealed three clusters of CCEV in Ethiopia with the isolates clustering according to their geographical origins. To our knowledge, this is the first report indicating the existence of CCEV in Ethiopia where camel contagious ecthyma was misdiagnosed as camelpox. Additionally, this study has also disclosed the existence of co-infections with CMLV and CCEV. A comprehensive characterization of poxviruses affecting camels in Ethiopia and the full genome sequencing of representative isolates are recommended to better understand the dynamics of pox diseases of camels and to assist in the implementation of more efficient control measures.

Molecular characterization of orf virus from sheep and goats in Ethiopia, 2008-2013.

BACKGROUND: Orf is a contagious disease of sheep, goats and wild ungulates caused by orf virus (ORFV) a member of the genus Parapoxvirus, Poxviridae family. Although orf is endemic in Ethiopia, little attention has been given so far as it is not a notifiable disease by the World Organization for Animal Health. In this work, we have investigated orf outbreaks representing five different geographical locations of Ethiopia, in Amba Giorgis, Gondar zuria, Adet, Debre zeit and Adami Tulu, between 2008 and 2013. RESULTS: The viral isolation and the sequence analysis of the A32L and the B2L genes of eighteen representative isolates confirmed that sampled animals were infected by ORFVs. The phylogenetic study and the comparative analysis of the deduced amino acid profile suggests that there were two main clusters of ORFV isolates which were responsible for the investigated outbreaks. Additionally the analysis of these two genes showed limited variability to ORFVs encountered elsewhere. This is the first report on the genetic characterization of the ORFV isolates from sheep and goats in Ethiopia. CONCLUSION: The molecular characterization of Ethiopian ORFV isolates highlighted the circulation of two main clusters causing orf disease in sheep and goats. The use of laboratory based methods and a constant monitoring of Ethiopian ORFV isolates is needed to better understand the dynamic of ORFV circulating in the country and facilitate the implementation of control measures.

A meta-analysis of contagious caprine pleuropneumonia (CCPP) in Ethiopia.

This systematic literature review was initiated due to lack of comprehensive information on the status and distribution of contagious caprine pleuropneumonia (CCPP) in Ethiopia. The objectives of the review were thus to provide a pooled prevalence estimate of CCPP in the country and asses the level of in between study variance among the available reports. Manual and electronic search was conducted between 8th of January and 25th of June 2015. A total of twelve published articles and one MSc thesis was retrieved from 19 initially identified studies. Twenty five animal level datasets were extracted at regional level considering some hypothesized predictors. The retrieved data were summarized in a meta-analytical approach. Accordingly, the pooled prevalence estimate of CCPP was 25.7% (95% CI:20.9,31.0). The inverse variance square (I(2)) that explains the variation in effect size attributed to reports true heterogeneity was 95.7%.The sub-group analysis was also computed for assumed predictors including, age, sex, type of study population, production systems and regional states. Among these predictors, study population type revealed statistically significant difference (P<0.05). Accordingly, the prevalence estimate for samples collected at abattoir was 39.2%, while that of samples collected at field level was 22.4%. In the final model, type of study population fitted the multivariable meta-regression model accounting for 22.87% of the explainable proportion of heterogeneity among the presumed predictors. Evidence on isolation and confirmation of Mycoplasma capricolum subspp. capripneumonie in the country was obtained from five regional states. In conclusion, it is recommended to further investigate facilities related with transportation and collection premises along with potential role of sheep in the epidemiology of CCPP. Finally, the review emphasizes the need for monitoring the ongoing CCPP control intervention and introduces amendments based on the findings. Besides more surveys are needed in some of the regions where no or few valid data was available.

Evaluation of the safety, immunogenicity and efficacy of three capripoxvirus vaccine strains against lumpy skin disease virus.

The safety, immunogenicity and efficacy of three commercially available vaccines against lumpy skin disease (LSD) in cattle have been evaluated using a combination of vaccine challenge experiments and the monitoring of immune responses in vaccinated animals in the field. The three vaccines evaluated in the study included two locally produced (Ethiopian) vaccines (lumpy skin disease virus (LSDV) Neethling and Kenyan sheep and goat pox (KSGP) O-180 strain vaccines) and a Gorgan goat pox (GTP) vaccine manufactured by Jordan Bio-Industries Centre (JOVAC). The latter vaccine was evaluated for the first time in cattle against LSDV. The Ethiopian Neethling and KSGPO-180 vaccines failed to provide protection in cattle against LSDV, whereas the Gorgan GTP vaccine protected all the vaccinated calves from clinical signs of LSD. There was no significant difference in protective efficacy detected between two dosage levels (P=0.2, P=0.25, and P=0.1 for KSGP, Neethling and Gorgan vaccines, respectively). Additionally, the Gorgan GTP vaccinated cattle showed stronger levels of cellular immune responses measured using Delayed-Type Hypersensitivity (DTH) reactions at the vaccination site indicating higher levels of immunogenicity produced by the GTPV vaccine in cattle, as opposed to the other two vaccines. This study indicated, for the first time, that the Gorgan GTP vaccine can effectively protect cattle against LSDV and that the Neethling and KSGP O-180 vaccine were not protective. The results emphasise the need for molecular characterization of the Neethling and KSGP O-180 vaccine seed viruses used for vaccine production in Ethiopia. In addition, the potency and efficacy testing process of the Ethiopian LSD Neethling and KSGP O-180 vaccines should be re-evaluated.

Capripox disease in Ethiopia: Genetic differences between field isolates and vaccine strain, and implications for vaccination failure.

Sheeppox virus (SPPV), goatpox virus (GTPV) and lumpy skin disease virus (LSDV) of the genus Capripoxvirus (CaPV) cause capripox disease in sheep, goats and cattle, respectively. These viruses are not strictly host-specific and their geographical distribution is complex. In Ethiopia, where sheep, goats and cattle are all affected, a live attenuated vaccine strain (KS1-O180) is used for immunization of both small ruminants and cattle. Although occurrences of the disease in vaccinated cattle are frequently reported, information on the circulating isolates and their relation to the vaccine strain in use are still missing. The present study addressed the parameters associated with vaccination failure in Ethiopia. Retrospective outbreak data were compiled and isolates collected from thirteen outbreaks in small ruminants and cattle at various geographical locations and years were analyzed and compared to the vaccine strain. Isolates of GTPV and LSDV genotypes were responsible for the capripox outbreaks in small ruminants and cattle, respectively, while SPPV was absent. Pathogenic isolates collected from vaccinated cattle were identical to those from the non-vaccinated ones. The vaccine strain, genetically distinct from the outbreak isolates, was not responsible for these outbreaks. This study shows capripox to be highly significant in Ethiopia due to low performance of the local vaccine and insufficient vaccination coverage. The development of new, more efficient vaccine strains, a GTPV strain for small ruminants and a LSDV for cattle, is needed to promote the acceptance by farmers, thus contribute to better control of CaPVs in Ethiopia.

Molecular evolution of peste des petits ruminants virus.

Despite safe and efficacious vaccines against peste des petits ruminants virus (PPRV), this virus has emerged as the cause of a highly contagious disease with serious economic consequences for small ruminant agriculture across Asia, the Middle East, and Africa. We used complete and partial genome sequences of all 4 lineages of the virus to investigate evolutionary and epidemiologic dynamics of PPRV. A Bayesian phylogenetic analysis of all PPRV lineages mapped the time to most recent common ancestor and initial divergence of PPRV to a lineage III isolate at the beginning of 20th century. A phylogeographic approach estimated the probability for root location of an ancestral PPRV and individual lineages as being Nigeria for PPRV, Senegal for lineage I, Nigeria/Ghana for lineage II, Sudan for lineage III, and India for lineage IV. Substitution rates are critical parameters for understanding virus evolution because restrictions in genetic variation can lead to lower adaptability and pathogenicity.

Meta-analysis of Brucella seroprevalence in dairy cattle of Ethiopia.

This meta-analysis estimates a single-group summary (effect size) for seroprevalence of Brucella spp. exposure in dairy cattle of Ethiopia. It also attempts to identify study-level variables that could explain the variation in apparent seroprevalence. The literature search was restricted to studies published in English language from January 2000 to December 2013. A template was designed to retrieve the most biologically plausible and consistent variables from the articles. A total of 29 published papers containing 40 animal-level studies were used in the analyses. The single-group summary of Brucella seroprevalence in cattle was estimated to reach 3.3 % with 95 % confidence interval (CI) (2.6-4.2 %). Of all the variables considered, region was the only specific factor identified to explain about 20 % of between-study variation. Accordingly, the region-based meta-analysis forest plot revealed the highest prevalence in central Ethiopia followed by southern part. The lowest prevalence estimate was observed in the western part of the country. The visual inspection of the funnel plot demonstrated the presence of possible publication bias which might dictate shortage of studies with higher prevalences or variance inflation due to infectiousness of Brucella. In conclusion, the quantitative review showed the seroprevalence to be low but widely distributed. More importantly, the review underscores the need for isolation and characterization of the circulating Brucella spp. to capture the type of Brucella spp. involved and its distribution in cattle in Ethiopia.

Sequence variability in the structural protein-encoding region of foot-and-mouth disease virus serotype A and O of Ethiopian isolates.

A total of 13 serotype O and 5 serotype A FMD Ethiopian isolates and some isolates from other countries (six for serotype A and four for serotype O) were sequenced on the structural protein (P1) coding region. The deduced amino acid sequences were aligned and investigated in an attempt to determine the amino acid variation. Differences were observed at 115 (15.6%) and 119 (16.1%) amino acid positions for serotype O and serotype A, respectively. The variation in the derived amino acid sequences is the highest in VP1, while VP4 was highly conserved in both serotypes A and O. In all isolates, hypervariable regions were located at regions corresponding to the highly immunogenic sites, the G-H loop (133-158) and the C-terminus (194-213) of the VP1 gene. The RGD cell attachment site within the G-H loop of the gene was conserved in all isolates. The study revealed the presence of significant amino acid variation at VP2 and VP3 in addition to known VP1 coding region. Hence, determination of amino acid sequence of the whole P1 region provides more information on antigenic variability of FMD virus and could be used in vaccine strain selection in parallel with serological vaccine matching assays.

Genetic basis of antigenic variation in foot-and-mouth disease serotype A viruses from the Middle East.

Foot-and-mouth disease viruses (FMDV) from serotype A exhibit high antigenic diversity. Within the Middle East, a strain called A-Iran-05 emerged in 2003, and subsequently replaced the A-Iran-96 and A-Iran-99 strains that were previously circulating in the region. Viruses from this strain did not serologically match with the established A/Iran/96 vaccine, although most early samples matched with the older A22/Iraq vaccine. However, many viruses from this strain collected after 2006 had poor serological match with the A22/Iraq vaccine necessitating the development of a new vaccine strain (A/TUR/2006). More recently, viruses from the region now exhibit lower cross-reactivity with the A/TUR/2006 antisera highlighting the inadequacy of the serotype A vaccines used in the region. In order to understand the genetic basis of these antigenic phenotypes, we have determined the full capsid sequence for 57 Middle Eastern viruses isolated between 1996 and 2011 and analysed these data in context of antigenic relationship (r1) values that were generated using antisera to A22/Iraq and A/TUR/2006. Comparisons of capsid sequences identified substitutions in neutralising antigenic sites (1, 2 and 4), which either individually or together underpin these observed antigenic phenotypes.

Genetic characterisation of infectious bursal disease virus isolates in Ethiopia.

The objective of the investigation was to characterise infectious bursal disease viruses (IBDV) circulating in commercial and breeding poultry farms in Ethiopia between 2009 and 2011. The nucleotide and deduced amino acid sequence for VP2 hypervariable region of ten IBDVs were determined by RT-PCR, sequenced and compared to well characterised IBDV isolates worldwide. IBDV genetic material was amplified directly from bursa or cell passaged material. Phylogenetically, Ethiopian IBDVs represented two genetic lineages: very virulent (vv) IBDVs or variants of the classical attenuated vaccine strain (D78). The nucleotide identity between Ethiopian vvIBDVs ranged between 0% and 2.6%. Ethiopian vvIBDVs are clustered phylogenetically with the African IBDV genetic lineage, independent of the Asian/European lineage. This report demonstrates the circulation of vvIBDV in commercial and breeding poultry farms in Ethiopia.

The first isolation and molecular characterization of camelpox virus in Ethiopia.

A cross-sectional study was conducted from November 2011 to April 2012 in Chifra district of Afar and in Jigjiga Zone of Somali Regional States of Ethiopia with the aims of assessing the epidemiology of camelpox and isolate and molecularly characterize the virus. The study included a questionnaire, active disease search and virus isolation and sequencing. A total of 24 (4.50%) and 12 (3.0%) camels in Afar and Jigjiga respectively were found clinically sick of camelpox during the study period. The questionnaire survey indicated that camelpox is the most common disease in the areas in which 125 (96%) of the respondents reported the frequent occurrence of camelpox in their herds especially during rainy season. The PCR result revealed 12 out of 17 tested samples were positive, of which seven of them collected from Jigjiga zone showed the characteristic PCR positive bands of 881 bp size fragments while five of the Afar samples gave two faint bands. Ethiopian isolates, specially isolated from Somali have very high identity with comparable sequences of CMLV M-96 from Kazakhstan and CMLV CMS from Iran. Out of the total of 780 bp analogous sequences, Ethiopian isolates differ only in two positions, while CMLV-Teheran differed at four nucleotide positions. The successfull isolation and molecular characterization of camelpox virus in Ethiopia, which could help for early diagnosis and control of the disease in the country.

Outbreak investigation and molecular characterization of African horse sickness virus circulating in selected areas of Ethiopia.

The study was conducted from June 2011 to May 2012 in central, northern and western parts of Ethiopia to investigate and identify circulating serotypes of African horse sickness virus (AHSV). The indigenous knowledge of equine owners about AHS in the study areas was assessed and also the retrospective data of AHS outbreaks for 2011 were analyzed. Whole blood samples were collected for virus isolation and serotyping from diseased horses and mules showing typical signs of the AHS. Virus isolation on Vero cell and detection of AHSV genomes using conventional RT-PCR were conducted. Further molecular characterization and serotyping were done on positive isolates. The questionnaire survey revealed that equine owners do recognize AHS clinically and have a local name that varies in different regions. From the 72 equine owners interviewed about their knowhow of AHS, 48 (66.7%) of respondents were not aware of AHS disease mode of transmission. The retrospective disease report data showed that a total of 208 outbreaks were reported and 3036 cases and 1167 deaths were recorded in 2011. AHS outbreaks were more frequently observed from September to December and the highest number of outbreaks was recorded in October. During the study period totally six outbreaks were investigated and a total of 62 horses and 10 mules were found sick and all the four forms of AHS were observed. Cardiac form accounted for 52.8%, followed by African horse sickness fever form 31.9%, pulmonary form 8.4% and mixed form 6.9%. AHSV-9 was the only serotype circulating in the outbreak areas.