Seroprevalence and molecular detection of foot and mouth disease virus in cattle in selected districts of Wolaita Zone, Southern Ethiopia.

Foot and mouth disease (FMD) is a highly contagious, endemic, and acute viral cattle ailment that causes major economic damage in Ethiopia. Although several serotypes of the FMD virus have been detected in Ethiopia, there is no documented information about the disease's current serostatus and serotypes circulating in the Wolaita zone. Thus, from March to December 2022, a cross-sectional study was conducted to evaluate FMDV seroprevalence, molecular detection, and serotype identification in three Wolaita Zone sites. A multistage sample procedure was used to choose three peasant associations from each study region, namely Wolaita Sodo, Offa district, and Boloso sore district. A systematic random sampling technique was employed to pick 384 cattle from the population for the seroprevalence research, and 10 epithelial tissue samples were purposefully taken from outbreak individuals for molecular detection of FMDV. The sera were examined using 3ABC FMD NSP Competition ELISA to find antibodies against FMDV non-structural proteins, whereas epithelial tissue samples were analyzed for molecular detection using real-time RT-PCR, and sandwich ELISA was used to determine the circulating serotypes. A multivariable logistic regression model was used to evaluate the associated risk variables. The total seroprevalence of FMD in cattle was 46.88% (95% CI 41.86-51.88), with Wolaita Sodo Town having the highest seroprevalence (63.28%). As a consequence, multivariable logistic regression analysis revealed that animal age, herd size, and interaction with wildlife were all substantially related to FMD seroprevalence (p < 0.05). During molecular detection, only SAT-2 serotypes were found in 10 tissue samples. Thus, investigating FMD outbreaks and identifying serotypes and risk factors for seropositivity are critical steps in developing effective control and prevention strategies based on the kind of circulating serotype. Moreover, further research for animal species other than cattle was encouraged.

Seroprevalence and associated risk factors of equine herpesvirus type-1/-4 in selected districts of Northwest Amhara, Ethiopia.

This study aimed to estimate the prevalence, determine the distribution, and identify the epidemiological risk factors of EHV-1/-4 infections in selected districts of Northwest Amhara Region. 460 serum samples were collected from equines using multistage cluster sampling technique, and a competitive Enzyme-linked immunosorbent assay (cELISA) was performed. Various risk factors for the occurrence of EHV-1/-4 were considered. Statistical analysis was performed using R version 4.3.1. 65.9% (303) equids were tested positive for antibodies against EHV-1/-4. Based on district, the highest prevalence was recorded in Wogera (86.1%), while the lowest was in Debark (47.4%). There was a significant difference (p <0.05; 95% CI: 1.1067993-3.682843) in the prevalence of EHV-1/-4 among species and donkeys are 2.019 times more likely to get an EHV infection than horses. The prevalence of EHV-1/-4 was highest in equids with the age of 3-8 years and lowest in < 3 years, and the difference was statistically significant (p <0.05; 95% CI: 1.9812042-6.771820). Statistically significant variation (p <0.05; 95% CI: 1.1173822-2.684013) was also observed between sex of equids in which females had 1.73 times higher chance to get EHV infection than males. Higher prevalence was found in lactating equids (81.6%), followed by pregnant equids (74.6%), and dry equids (66.4%). Generally, this study indicated a high and wide distribution of EHV-1/-4 infection in the study area, which needs due attention. Devising strategies to prevent and minimize the spread and occurrence of the infection is crucial.

Humoral Immune Response in Calves Vaccinated with Monovalent Vaccines or a Trivalent Combination Thereof and Matching of These Vaccines to the Selected Circulating Foot-and-Mouth Disease Viruses in Ethiopia.

Foot-and-mouth disease (FMD) is an endemic, highly contagious, and devastating disease of livestock production in Ethiopia. Control of this disease relies mainly on prophylactic vaccination by willing farmers without a countrywide vaccination program. The objectives of this study were to quantify the humoral immune response and evaluation of the serological relationship of the vaccine strain used with representative field strain isolates. This was performed by primo vaccination of 6-9-month-old Holstein Friesian calves (35 treatment and 4 control calves) on day one and booster vaccination on day 28. Calves were vaccinated using the locally available National Veterinary Institute (NVI), Bishoftu, Ethiopia, inactivated aluminum hydroxide adjuvant monovalent (either O, A, SAT-2 alone) or trivalent (combination of A, O, SAT-2) vaccine (A/ETH/6/2000 (G-VII, O/ETH/38/2005(EA-3) and SAT-2/ETH/64/2009(XIII)). A 2 mL or 4 mL dose was used to vaccinate all calves except the animals that served as a control. In the case of the trivalent vaccine, a 4 mL dose was used to vaccinate calves. The serum was collected at 7, 14, 21, 28, and 56 days post-vaccination (d.p.v.). The humoral immune response was quantified by the solid-phase competitive enzyme-linked immunosorbent assay (SPC ELISA) and the virus-neutralization test (VNT). The serological relationship of heterologous and homologous viruses was also evaluated by adjuvant vaccine matching tests. The r1-value was determined using serum collected 21 d.p.v. An increase in immune response was observed from 7 d.p.v. to 28 d.p.v. in calves who received a 4 mL dose containing a 107.24 antigen load of 100 tissue culture infective dose (100TCID50) virus titer in the formulation. Upon receiving a booster dose on day 28, the humoral immune response was checked on the 56th day post-initial vaccination. Amounts of 54%, 72%, 79%, and 72% of inhibition for A, O, SAT-2, and trivalent vaccine in the three serotypes SPCE, respectively, was measured. Here, it was found that the immune response of calves increased from day 7 to 56, as evidenced by SPCE analysis. Likewise, an increase in antibody titer measured by a one-dimensional virus neutralization assay was also in line with SPCE analysis. This indicates that the vaccine is capable of inducing a neutralizing antibody that confers a protective immune response in 70%, 62%, and 100% heterologous field strains of A, O, and SAT-2 isolates, respectively, and has an average antigenic relationship of >0.3 with a standard deviation of +0.05 (N = 3) to the vaccine strains A/ETH/6/2000, O/ETH/38/2005 and SAT-2/ETH/64/2009, respectively, when using the one-dimensional virus neutralization test. The contribution and importance of this study is a confirmation of the vaccine and the field strain serological relationship for serotype SAT-2 strain and further research/change of vaccination strategy/ improvement in the currently used vaccine to cover a wide range of prevailing genotypes/lineages and induction of sound immune response after vaccination for serotype A and O strain. This study suggests that the trivalent vaccine produced by the National Veterinary Institute containing viral isolates from serotype O, A, and SAT-2 has a good serological relationship with the majority of circulating field strains in Ethiopia.

Epidemiological study and dairy farmers' knowledge, attitudes, and practices on foot and mouth disease in central Ethiopia.

Foot and mouth disease (FMD) is consistently ranked as the most economically significant viral disease and one of the top five livestock diseases in Ethiopia. Although FMD is endemic in Ethiopia, the epidemiology and the farmers' knowledge, attitudes, and practices regarding FMD were poorly quantified. Thus, a cross-sectional study was conducted from November 2021 to April 2022 to estimate the seroprevalence, identify the FMD serotypes, and assess the farmers' knowledge, attitudes, and practices on FMD in Addis Ababa city and Sebeta special zone, central Ethiopia. A total of 384 serum samples were collected from cattle and tested using a 3ABC enzyme-linked immunosorbent assay (ELISA). In this study, an overall 56% seroprevalence was recorded. Two types of FMD serotypes were detected in which serotype O was the dominant serotype (75.5%) followed by serotype A (45.5%). A significantly higher seroprevalence (P = 0.00) was recorded in Addis Ababa (85%) compared to Sebeta (28.7%). Seropositivity in older and semi-intensively managed cattle was 2.9 (95% CI: 1.36-6.50; P = 0.006) and 2.1 (95% CI: 1.34-3.26; P = 0.001) times higher compared to young and intensively managed cattle, respectively. A survey on knowledge, attitude, and practice of 103 farmers revealed that 90.2% knew of FMD and the majority of them can recognize its clinical pictures. However, 12.7% of farmers who knew FMD didn't practice any prevention methods. Additionally, 70% of the farmers responded that their cattle roamed outside of their farms for communal grazing, watering, breeding purposes, and vaccination which might put them more at risk of FMD. The current study demonstrated that the majority of farmers have gaps in biosecurity practices and vaccination of cattle against FMD. Therefore, educating farmers on FMD prevention measures is necessary for successful disease control programs.

Combined Adjuvant Formulations Enhanced an Immune Response of Trivalent Foot and Mouth Disease Vaccine in Cattle.

Introduction: Foot-and-mouth disease is globally one of the most economically important viral diseases of cloven-hoofed animals that can be controlled by different strategies, where vaccination plays an important role. Selection of adjuvant added to vaccine preparation is crucial in ensuring the protective effect of the vaccine. Aluminum hydroxide gel mixed with saponin (AS) is widely used adjuvant, with its suboptimal immune response in FMD vaccine. The present study was undertaken to evaluate different ingredients of adjuvants for inactivated trivalent (A, O and SAT 2) FMD vaccine and to demonstrate the effect of booster dose in cattle. Methods: Cattle were grouped into five; four experimental and one control, with six animals in each group and immunized with trivalent vaccine with various formulations of adjuvants. Immune response was measured using Solid Phase Competitive Enzyme Linked Immune Sorbent Assay (SPCE). Results: The antibody level in cattle immunised with a vaccine formulation containing a mixture of aluminum hydroxide gel and saponin (AS) were significantly lower than AS boosted group for the three serotypes (p<0.05, t-test), which directs the need for booster dose. Whereas the antibody response in the AS + oil group was higher followed by oil alone. The AS preparation with a booster dose has shown better immune response compared to the group without. Conclusion: The findings of this study could suggest that oil based and AS with oil could replace the conventional aluminum hydroxide gel and saponin adjuvants in FMD vaccine preparations. Challenge test was not successful indicating the need for further research on the virus infectivity.

Evaluation of diagnostic performance of H-based blocking ELISA for specific detection of peste des petits ruminants in domestic sheep, goats, cattle and camels.

INTRODUCTION: Peste des petits ruminants virus (PPRV) causes a highly devastating disease of sheep and goats, peste des petits ruminants (PPR), which is targeted for global control and eradication by 2030. The serological diagnostic tool kits for accurate diagnosis of PPR have inherent strengths and weaknesses that require parallel validation and optimization across animal species. Thus, the objective of this study was to evaluate diagnostic performance of haemagglutinin based PPR blocking ELISA (HPPR- b-ELISA), that was developed by Africa Union Pan African Veterinary Vaccine Center for specific detection of anti- PPRV antibodies. METHODS: In preliminarily investigation, diagnostic performance of the HPPR-b-ELISA®, commercial PPR competition ELISA (c-ELISA) and virus neutralization test (VNT) were compared for the detection of anti-PPRV antibodies in goats, sheep, cattle and camels. RESULTS: The sensitivity and specificity of HPPR- b-ELISA® were 79.55 and 99.74%, respectively, compared to c-ELISA. The HPPR- b-ELISA® was in perfect agreement (κ = 0.86) with the c-ELISA in all sera collected from goats, sheep and cattle. There was almost perfect agreement between the species of goats (κ = 0.82) and sheep (κ = 0.98), while the agreement was substantial in cattle (κ = 0.78) and no agreement was observed in camels (κ = 0.00). Similarly, the sensitivity and specificity of the HPPR b-ELISA were 80 and 96.36%, respectively compared to VNT with almost perfect agreement in goats (κ = 0.83) and sheep (κ = 0.89), moderate in cattle (κ = 0.50) and none in camels (κ = 0.00). CONCLUSION: Our study revealed that HPPR- b-ELISA is a suitable and valid method that can alternatively be used for screening and monitoring of PPR in sheep, goats and cattle except for camels.

Seroprevalence and associated risk factors for foot and mouth disease virus seropositivity in cattle in selected districts of Gamo zone, Southern Ethiopia.

Background: Foot and mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed animals, which hampers livestock production and productivity in Ethiopia. This cross-sectional study was conducted from January to December 2021 to estimate the seroprevalence of FMD in cattle and to assess farmers' knowledge about the disease in selected districts of the Gamo zone. Three districts and two kebeles (smallest administrative division) from each district were purposively sampled using a simple random sampling technique to select individual animals from each kebeles. A total of 384 sera samples were collected, and concurrently, 100 farmers were interviewed. The samples were tested for antibodies against nonstructural proteins of the FMD virus using a 3ABC enzyme-linked immunosorbent assay (ELISA). Univariable and multivariable logistic regressions were used to analyze FMD-associated risk factors. Result: The questionnaire survey result revealed that among the interviewed farmers, 66% of farmers had knowledge about the disease, and 28% of farmers reported having a case of FMD in at least one cattle in their farm in the previous 6 months. The overall seroprevalence of FMD in cattle was 26.8%. The multivariable logistic regression revealed that age, breed, and agroecology had a significant association with seropositivity. Higher seroprevalence (64.57%) was observed in lowland, followed by midland (9.30%) and highland (5.88%). Study animals from lowland areas were 9.26 times more likely to be seropositive (OR = 9.26, CI = 2.22-38.62) for FMD than highland animals. Also, adult animals were 9.01 times (OR = 9.01, CI = 3.18-25.53) more likely to be seropositive for the disease than young animals. The multivariable logistic regression revealed that crossbreeds have an 84.7% (OR = 0.153, CI = 0.028-0.82) lower likelihood to be seropositive to FMD than local breeds. Conclusion: This study result confirms that FMD is highly prevalent in the study area, and farmers' knowledge regarding disease transmission and vaccine availability is minimal. Hence the regional concerned bodies should implement FMD vaccination campaigns and create awareness for smallholder farmers regarding the disease transmission, FMD vaccine schedule, and vaccination importance.

Molecular characterization of foot-and-mouth disease viruses circulating in Ethiopia between 2008 and 2019.

One of the constraints to controlling foot-and-mouth disease (FMD) in East Africa is the incomplete knowledge of the specific FMD virus (FMDV) strains circulating and the way in which these viruses move across countries in the region. This retrospective study focuses on Ethiopia, which has one of the largest FMD-susceptible livestock populations in Africa. Analyses of FMDV positive samples collected between 2008 and 2019 demonstrate that serotypes O (n = 175), A (n = 51) and SAT 2 (n = 33) were present in the country. Phylogenetic analysis of the VP1 sequences for these viruses showed that there were at least seven different FMD viral clades circulating during this period: O/EA-3, O/EA-4, A/AFRICA/G-I, A/AFRICA/G-IV, A/AFRICA/G-VII, SAT2/VII and SAT2/XIII. Although these results only represent a snapshot and might not reflect all FMDV lineages that were present, they highlight the importance of serotype O, as well as the complexity and co-existence of FMDV serotypes in Ethiopia and surrounding countries. These sequence data also support the idea that there are two FMDV ecosystems existing in East Africa. Data from retrospective studies, such as these presented here, will be beneficial for vaccine selection and vaccination campaigns to control FMDV within Ethiopia.