Isolation and Characterization of Coronavirus and Rotavirus Associated with Calves in Central Part of Oromia, Ethiopia.

BACKGROUND: Coronavirus and rotavirus are most commonly associated etiologies for calves' diarrhoea, resulting in loss of productivity and economy of farmers. However, various facets of diarrheal disease caused by coronavirus and rotavirus in calves in Ethiopia are inadequately understood. A cross-sectional study was conducted with the aim of isolation and molecular characterization of coronavirus and rotavirus from calves in the central part of Oromia (Bishoftu, Sebata, Holeta, and Addis Ababa), Ethiopia, from November 2018 to May 2019. The four study areas were purposively selected and faecal samples were collected by simple random sampling for diagnosis of coronavirus and rotavirus infection by using the antigen detection enzyme-linked immunosorbent assay (Ag-ELISA) kit. In addition, this study was carried out to have insight in prevalence and associated risk factors of coronavirus and rotavirus infection in calves. RESULT: During the study, 83 diarrheic and 162 nondiarrheic faecal samples collected from calves less than 4 weeks of age were screened for coronavirus and rotavirus. Of the 83 diarrheic samples, 1 sample (1.2%) was positive for coronavirus antigen and 6 samples (7.2%) were found to be positive for rotavirus antigen by Ag-ELISA. All the nondiarrheic samples were negative for both coronavirus and rotavirus Ag. The overall prevalence of coronavirus and rotavirus infection in calves was estimated at 0.4% (1/245) and 2.45% (6/245), respectively. All samples (7) of ELISA test positive of both coronavirus and rotavirus were propagated in Madin-Darby bovine kidney (MDBK) cells. After 3 subsequent passages, progressive cytopathic effect (CPE), i.e., rounding, detachment, and the destruction of monolayer cell of five samples (1 sample of coronavirus and 4 samples of rotavirus) (71.4%) were observed. At the molecular stage, reverse transcriptase polymerase chain reaction (RT-PCR) technique was used to determine the presence of coronavirus and rotavirus nucleic acid by using specific primers. The 5 samples that were coronavirus and rotavirus antigen positive by ELISA and develop CPE on cell culture were also positive on RT-PCR technique. The prevalence of infection peaked at 1st and 2nd weeks of age in male calves. CONCLUSION: Diarrheal disease caused by coronavirus and rotavirus has a great health problem in calves that interrupts production benefits with reduced weight gain and increased mortality and its potential for zoonotic spread. So, the present findings show coronavirus and rotavirus infection in calves in Ethiopia that needs to be addressed by practising early colostrum feeding in newborn calves, using vaccine, or improving livestock management.

Isolation, molecular characterization and sero-prevalence study of foot-and-mouth disease virus circulating in central Ethiopia.

BACKGROUND: Ethiopian livestock production and productivity is still very low due to widespread of diseases. Among the diseases, foot-and-mouth disease (FMD) is an extremely contagious and acute viral disease that causes significant economic problems in the country. A cross sectional study design was conducted from September 2015 to May 2016 to isolate and characterize FMD virus from outbreak cases; determine the sero-prevalence of antibodies against FMD virus (FMDV), and assess potential risk factors associated with sero-prevalence of the disease in selected areas of central Ethiopia. A multistage sampling technique was employed to select the study animals. Isolated viruses were characterized by antigen ELISA (IZLER, Brescia, Italy) and by genetic analysis of the sequence of the viral protein 1 (VP1). Sero-prevalence was determined using an ELISA for antibodies against non-structural proteins of FMDV based on the 3ABC proteins (ID Screen® FMD NSP Competition, ID-VET, Grabels, France). Risk factors for sero-prevalence of antibodies against FMD virus was investigated using logistic regression analysis. RESULT: From outbreak investigation, 28.8% (n = 378) cattle showed signs and lesions suggestive of FMD and 34 samples were subjected to virus isolation. Twenty eight of these cultures exhibited cytopathic effect (CPE) and were serotyped as O, A and SAT 2 FMD viruses. One A and two SAT 2 isolates named A-ETH-19-2015, SAT 2-ETH-18-2015 and SAT 2-ETH-20-2015 were further characterized by phylogenetic analysis. The overall sero-prevalence of antibodies against non-structural proteins of FMDV was 24.2% (n = 574). Cattle herds with crossbreed cattle, with older cattle (> 2 years), and kept together with small ruminants had higher sero-prevalences of antibodies against FMDV (p < 0.05). CONCLUSION: This study showed that FMD was present in the study areas. Among the associated risk factors, breed, age and herd composition were significantly associated with presence of antibodies against FMD virus. Three different serotypes (A, O and SAT 2) were responsible for the outbreaks of the disease. Genetic analysis indicated that the isolated viruses clustered differently from previous outbreaks. Thus, further molecular analyses coupled with protection potential of the existing vaccines against the isolates should be performed.

Aerobic bacteriological studies on the respiratory tracts of apparently healthy and pneumonic camels (Camelus dromedaries) in selected districts of Afar Region, Ethiopia.

A cross-sectional study was conducted to isolate and identify bacterial species from the respiratory tract of apparently healthy and pneumonic camels in Asayita and Dubti woredas in the Afar Region, Ethiopia. From a total of 74 lung tissue and 74 tracheal swab samples Staphylococcus aureus, 16.3%, Streptococcus equi subsp. equi, 13.0%, and Pasteurella multocida, 10.9%, were dominant isolates from pneumonic lungs; Escherichia coli, 12.7%, Proteus species, 10.9%, and Klebsiella pneumoniae, 9.1%, were the majority in the normal lungs. The majority of the isolates colonized both anatomical sites investigated. There was a statistically significant association between the health status of the camels as well as the anatomical site studied with the isolation rates of the major respiratory pathogens (p < 0.05). Furthermore, the isolates were susceptible to norfloxacin, streptomycin, and gentamicin but resistant to ampicillin and tetracycline on in vitro test. Further studies on the pathogenicity of the major isolates are recommended.

Toxoplasmosis in camels (Camelus dromedarius) of Borana zone, Oromia region, Ethiopia: seroprevalence and risk factors.

Toxoplasmosis is one of the most prevalent parasitic infections of medical and veterinary importance. A cross-sectional study was conducted from November 2013 to January 2014 to estimate the seroprevalence of Toxoplasma gondii infection in camels from four districts of Borana zone, Southern Oromia, Ethiopia. In addition, a questionnaire survey was administered to 124 pastoralists to identify possible risk factors and to assess the awareness level of pastoral communities about toxoplasmosis. A total of 396 serum samples were examined for anti-Toxoplasma IgG antibodies using the direct agglutination test (DAT). Fisher's exact test and logistic regression were used for data analysis. An overall seroprevalence of 8.33 % (95 % confidence interval [CI] 5.60 %, 11.07 %) at animal-level and 37.5 % (95 % CI: 20.1 %, 57.4 %) at herd-level was found. The seroprevalence was significantly high in Moyale district (23.07 %) followed by Yabello (7.20 %), Dirre (3.77 %), and Arero (0.0 %) districts (P < 0.001). Multivariable logistic regression analysis revealed that the likelihood of acquiring T. gondii infection was significantly higher in camels of Moyale district (adjusted OR = 5.89, 95 % CI 2.15, 16.12; P = 0.001) than Dirre district, in camels of >8 years old (adjusted odds ratio [OR] = 4.95, 95 % CI 1.68, 14.55; P = 0.004) than camels of ≤4 years old. There was no significant association between herd-level seroprevalence of T. gondii infection and abortion history, herd size, and presence of domestic cats and wild felids (P > 0.05). The majority of interviewees were uneducated (82.25 %), and all had no knowledge of toxoplasmosis. All camel herders drink raw camel milk but consume cooked meat (90.32 %). Of the interviewees, 93.06 % are aware about soil-eating habit of camels and provide salt supplement for their camels. Majority of the respondents practice improper disposal of aborted materials (throw along the way) (88.70 %), and 73 % of the study participants do not wash their hands after handling aborted fetus. The results of the present study confirm relatively lower prevalence of T. gondii infection in camels reared in Borana zone. Age and study district are significant predictors of T. gondii seropositivity. The vast majorities of interviewed pastoralists were uneducated and practice poor biosecurity measures to prevent diseases. Education of pastoralists about biosecurity measures to prevent toxoplasmosis and further studies are warranted to unravel the economic and public health consequences of T. gondii infection.

Serological survey of viral diseases relating to reproductive failure among Artiodactyla in Ethiopian Camelus dromedarius.

Serological surveys were performed on Ethiopian camels with a history of abortion to investigate the presence of antibodies against viruses that infect animals classified in the order Artiodactyla. In 2013, 120 serum samples were collected from camels in various parts of Ethiopia. Several viruses related to abortion in ruminants were prevalent. In particular, antibodies against bluetongue virus, were detected at a high rate (76.7% of samples). Additionally, antibodies against Akabane virus and Japanese encephalitis virus were also detected in samples from more than 40% of the camels; however, their antibody titers were relatively low.

First report of Toxoplasma gondii in camels (Camelus dromedarius) in Ethiopia: bioassay and seroepidemiological investigation.

BACKGROUND: Toxoplasmosis is a major public health concern in many countries of the world. A cross-sectional and follow up experimental study designs were used for seroepidemiological and bioassay studies, respectively from November 2012 to April 2013. The objectives were to estimate the seroprevalence of T. gondii infection, to assess risk factors and to isolate the parasite from camels in the Fentale district, Ethiopia. A direct agglutination test (DAT) and indirect enzyme linked immunosorbent assay (ELISA) kits were used to test camel sera. Hearts and tongues (each 25 g) from 31 seropositive camels were bioassayed in mice. Associations between seroprevalence and potential risk factors (collected using a questionnaire survey) were analyzed using logistic regression. RESULTS: An overall T. gondii prevalence of 49.62% (220/455) by DAT and 40.49% (179/451) by indirect ELISA test were detected. Herd level seroprevalence of 96.77% (30/31) (95% CI: 83.30- 99.92) by DAT was recorded and it was significantly higher in areas where wild felids are present (P = 0.038). Multivariable logistic regression showed that the likelihood of acquiring T. gondii infection was significantly higher in camels in the Ilala pastoral association [PA] (82.26%) (Adjusted Odds ratio [aOR] = 10.8; P < 0.001) than camels in the Galcha PA (31.43%), in camels of ≥ 8 years old (56.52%; aOR = 1.88; P = 0,033) than camels of ≤ 4 years old (34.26%) and in areas where domestic cats are present (aOR = 4.16; P = 0.006). All camel owners were uneducated, handle aborted fetus with bare hands, and drink raw camel milk. DAT and ELISA tests had moderate agreement (Kappa = 0.41). Viable T. gondii were isolated from 16.13% (5/31) of DAT positive camels. One DAT positive but ELISA negative camel sample gave a cyst positive result. CONCLUSIONS: T. gondii infection of camels in the study district is widespread. Age, presence of domestic cats and study PA are independent predictors of T. gondii seropositivity. Isolation of viable parasites from edible tissues of camels and the very poor knowledge of pastoralists about toxoplasmosis suggest the need for prevention of toxoplasmosis through bio-security measures, education and further investigation to unravel the impact of camel toxoplasmosis deserves consideration.

Geographic distribution of MERS coronavirus among dromedary camels, Africa.

We found serologic evidence for the circulation of Middle East respiratory syndrome coronavirus among dromedary camels in Nigeria, Tunisia, and Ethiopia. Circulation of the virus among dromedaries across broad areas of Africa may indicate that this disease is currently underdiagnosed in humans outside the Arabian Peninsula.

Use of immunoblotting to detect antibodies to Mycoplasma crocodyli infection in the sera of crocodiles (Crocodylus niloticus).

An immunoblotting protocol for the detection of antibodies to Mycoplasma crocodyli was developed using sonicated antigen of the reference strain 266/93. Immunoblotting detected nine reacting antigens, of which the 33 and 40kDa antigens were immunodominant. There was no difference in reactivity of the antigens against sera obtained from vaccinated and infected crocodiles. Both antigens are candidates for other serological and molecular studies. This is the first report to develop and apply an immunoblotting test for detection of antibody to M. crocodyli infection in crocodiles.

Development and application of an indirect ELISA test for the detection of antibodies to Mycoplasma crocodyli infection in crocodiles (Crocodylus niloticus).

Non-availability of a standardized rapid serodiagnostic test for quick and accurate diagnosis of Mycoplasma crocodyli (M. crocodyli) infection in crocodiles was the underlining reason for conducting the present study. An indirect enzyme-linked immunosorbent assay (iELISA) for the detection of antibodies (Ab) to M. crocodyli infection in crocodile sera was developed using sonicated antigen (Ag) and anti-crocodile conjugate. The iELISA test was optimised with different reagents and at different steps. A cut-off value of percent positive greater than or equal to 53.47% resulted in an estimated sensitivity and specificity of 85.67 and 100%, respectively. The developed iELISA could be used for detection of Abs to M. crocodyli infection in crocodiles and may enable to understand the transmission of the disease.