Epidemiological Survey of Peste des Petits Ruminants in Ethiopia: Cattle as Potential Sentinel for Surveillance.

Peste des petits ruminants (PPR) is a highly contagious viral disease of small ruminants; it emerged in countries previously free of the disease following the eradication of rinderpest. PPR is classified by international organizations as the next priority animal disease for global eradication campaign. Assessment of the local situations is the first step in the eradication efforts. The objective of this study was to investigate and compare the seroprevalence of PPR in cattle, sheep, and goats under two livestock production systems in Ethiopia: North Shewa zone of Amhara region represents a highland sedentary life style characterized by mixed livestock-crop production system; Zone Three of Afar region represents a lowland nomadic life style characterized by pastoral livestock production system. N-competitive ELISA PPR test was performed on sera from 2,993 animals ≥6 months old sampled at watering and grazing points. Multivariable logistic regression models comparing the seropositivity between the two production systems were built by classifying doubtful results as positive, negative, or excluding them from the data. The odds ratio (OR) comparing overall PPR seroprevalence in the sedentary North Shewa Zone compared to the nomadic Zone Three ranged from 19 to 27 (P < 0.001), depending on how doubtful results were classified, which contrasts with what has been reported in the literature. This is not likely to be related solely to vaccination, since seroprevalences in cattle and small ruminants were similarly high or low in the respective zones (0-4% for Zone Three and 20-40% for North Shewa Zone), and cattle were not likely to be vaccinated. The OR of seropositivity for goats compared to cattle ranged from 1.9 [95% confidence interval (CI): 1.3-2.7; P < 0.001] to 2.2 (95% CI: 1.5-3.1; P < 0.001) when doubtful results were excluded or classified as negative, respectively. When doubtful results were classified as positive, association between seropositivity and animal species was not significant (P > 0.05). Our results suggest to further investigate cattle as sentinel animals for PPR surveillance.

Histopathological lesions in reproductive organs, distal spinal cord and peripheral nerves of horses naturally infected with Trypanosoma equiperdum.

BACKGROUND: Dourine, a venereal transmitted trypanosomosis caused by Trypanosoma equiperdum, has different clinical signs related to the reproductive and nervous system. Pathologic tissue changes associated with the disease are poorly described. The present study describes the histopathological lesions in naturally T. equiperdum-infected horses in the chronical stage of dourine. RESULTS: Four chronically dourine diseased horses underwent a post-mortem examination. They were Woo test negative, but CATT/T. evansi positive, had a low packed cell volume (PCV) and exhibited obvious clinical signs of dourine. Post-mortem examination did not reveal gross lesions in the organs assumed to be responsible for the symptomatology. On histopathology, genital organs were affected, with mononuclear cell infiltration and erosions and degeneration of seminiferous tubules and perivascular lymphoplasmacytic cuffing in the uterus. In the nervous system, mononuclear cell infiltration was located in peripheral nerves, ganglia and in the spinal cord, leading to axonal degeneration. Real-time PCR using ITS primer revealed the presence of trypanosomes in these organs and conventional PCRs using maxicircle and RoTat1.2 primers further confirmed the involvement of T. equiperdum since the DNAs from the vagina, testicle, distal spinal cord, sciatic and obturator nerves found to be positive for maxicircle and negative for RoTat 1.2. CONCLUSIONS: The histopathological lesions in the spinal cord and peripheral nerves explain the incoordination of the hind legs in T. equiperdum-infected horses, whilst its presence in the genital tract exemplifies the venereal transmission.

Reduction of Trypanosoma equiperdum from equine semen by single layer centrifugation.

Trypanosoma equiperdum (T. equiperdum) causes dourine, a venereally transmitted infection in horses. Purification of semen by single layer centrifugation (SLC) has been proven to be successful in reducing venereally transmitted diseases when dealing with other pathogens. The objective of this study was to evaluate the purification of T. equiperdum spiked semen by SLC. Semen was spiked using cryopreserved T. equiperdum stabilates (Dodola strain isolate 943). In total, 6 concentrations, varying from 102 to >5 × 106 trypanosomes, were added to semen samples. Subsequently, SLC was performed following standard procedures. The presence of the parasite in the purified semen was checked by wet smear examination, ITS1 PCR and in vivo inoculation in mice. Before SLC, all spiked semen samples, except the negative controls, were positive on PCR analysis. After SLC, all the pellets were found to be negative for T. equiperdum on microscopic examinations. Examination of the pellet by PCR could also not detect any parasite-DNA in the SLC-pellet of semen spiked with the lower number of parasites (102 to104 trypanosomes). However, in the SLC pellets spiked with 104 - 5 × 104 trypanosomes, only 1 out of the 4 replicates was negative for parasite DNA. All groups spiked with >5 × 104 trypanosomes were found to be positive on PCR. All mice in the positive controls exhibited parasitaemia (5/5). Mice inoculated with SLC-purified semen that was spiked with lower than 5 × 104 trypanosomes, remained free of parasitaemia, similar to the negative controls. However inoculation with SLC-pellets from samples with a higher number of trypanosomes (>5 × 104 - 5 × 106 and > 5 × 106), induced parasitaemia in 2 out of 5 and 3 out of 5 mice, respectively. This study indicates that single layer centrifugation can be used to clear T. equiperdum infected semen but that the success is dependent on the number of parasites.

Tissue (re)distribution of Trypanosoma equiperdum in venereal infected and blood transfused horses.

Dourine, caused by Trypanosoma equiperdum, is a life-threatening venereal disease in equidae. So far, there is no clear evidence on how and when stallions become infectious, nor which tissues are affected by the parasite in diseased animals. Post-infection, after a transient, temporary phase of parasitaemia, the parasite disperses to different tissues in an unknown distribution pattern. This study describes the distribution of the parasite after infection by artificial insemination (AI) or blood transfusion. Mares (N = 4) were artificially inseminated with T. equiperdum spiked semen whereas stallions (N = 4) were infected by blood transfusion. The course of the disease was monitored by parasitological (Woo) and molecular (PCR) tests and clinical signs and haematological parameters were recorded. At 120 days post infection, horses had a full necropsy, histopathology and PCR. A similar pattern of parasitaemia, disease progression and tissue distribution were seen in all horses. Ejaculated semen in the preclinical stage and epididymal semen in the chronic stage of the disease was positive on PCR and caused infection in mice. Cymelarsan® treatment in the chronic stage did not result in a clinico-haematological or histopathological improvement. At necropsy, lesions were observed in the nervous and reproductive system. Histopathological lesions were most severe in the peripheral nerves and associated ganglia, the testicles and genital mucosae with multifocal infiltration of lymphocytes, plasma cells and histocytes. The parasites disseminated to several tissues including the nervous system, testicles and semen. The results indicate that transmission of T. equiperdum is possible through semen even from symptomless stallions post-treatment.

A systematic review and meta-analysis of trypanosome prevalence in tsetse flies.

BACKGROUND: The optimisation of trypanosomosis control programs warrants a good knowledge of the main vector of animal and human trypanosomes in sub-Saharan Africa, the tsetse fly. An important aspect of the tsetse fly population is its trypanosome infection prevalence, as it determines the intensity of the transmission of the parasite by the vector. We therefore conducted a systematic review of published studies documenting trypanosome infection prevalence from field surveys or from laboratory experiments under controlled conditions. Publications were screened in the Web of Science, PubMed and Google Scholar databases. Using the four-stage (identification, screening, eligibility and inclusion) process in the PRISMA statement the initial screened total of 605 studies were reduced to 72 studies. The microscopic examination of dissected flies (dissection method) remains the most used method to detect trypanosomes and thus constituted the main focus of this analysis. Meta-regression was performed to identify factors responsible for high trypanosome prevalence in the vectors and a random effects meta-analysis was used to report the sensitivity of molecular and serological tests using the dissection method as gold standard. RESULTS: The overall pooled prevalence was 10.3% (95% confidence interval [CI] = 8.1%, 12.4%) and 31.0% (95% CI = 20.0%, 42.0%) for the field survey and laboratory experiment data respectively. The country and the year of publication were found to be significantly factors associated with the prevalence of trypanosome infection in tsetse flies. The alternative diagnostic tools applied to dissection positive samples were characterised by low sensitivity, and no information on the specificity was available at all. CONCLUSION: Both temporal and spatial variation in trypanosome infection prevalence of field collected tsetse flies exists, but further investigation on real risk factors is needed how this variation can be explained. Improving the sensitivity and determining the specificity of these alternative diagnostic tools should be a priority and will allow to estimate the prevalence of trypanosome infection in tsetse flies in high-throughput.

Spatial distribution of Glossina sp. and Trypanosoma sp. in south-western Ethiopia.

BACKGROUND: Accurate information on the distribution of the tsetse fly is of paramount importance to better control animal trypanosomosis. Entomological and parasitological surveys were conducted in the tsetse belt of south-western Ethiopia to describe the prevalence of trypanosomosis (PoT), the abundance of tsetse flies (AT) and to evaluate the association with potential risk factors. METHODS: The study was conducted between 2009 and 2012. The parasitological survey data were analysed by a random effects logistic regression model, whereas the entomological survey data were analysed by a Poisson regression model. The percentage of animals with trypanosomosis was regressed on the tsetse fly count using a random effects logistic regression model. RESULTS: The following six risk factors were evaluated for PoT (i) altitude: significant and inverse correlation with trypanosomosis, (ii) annual variation of PoT: no significant difference between years, (iii) regional state: compared to Benishangul-Gumuz (18.0%), the three remaining regional states showed significantly lower PoT, (iv) river system: the PoT differed significantly between the river systems, (iv) sex: male animals (11.0%) were more affected than females (9.0%), and finally (vi) age at sampling: no difference between the considered classes. Observed trypanosome species were T. congolense (76.0%), T. vivax (18.1%), T. b. brucei (3.6%), and mixed T. congolense/vivax (2.4%). The first four risk factors listed above were also evaluated for AT, and all have a significant effect on AT. In the multivariable model only altitude was retained with AT decreasing with increasing altitude. Four different Glossina species were identified i.e. G. tachinoides (52.0%), G. pallidipes (26.0%), G.morsitans submorsitans (15.0%) and G. fuscipes fuscipes (7.0 %). Significant differences in catches/trap/day between districts were observed for each species. No association could be found between the tsetse fly counts and trypanosomosis prevalence. CONCLUSIONS: Trypanosomosis remains a constraint to livestock production in south-western Ethiopia. Four Glossina and three Trypanosoma species were observed. Altitude had a significant impact on AT and PoT. PoT is not associated with AT, which could be explained by the importance of mechanical transmission. This needs to be investigated further as it might jeopardize control strategies that target the tsetse fly population.

Widespread occurrence of Trypanosoma vivax in bovines of tsetse- as well as non-tsetse-infested regions of Ethiopia: a reason for concern?

A cross-sectional study was undertaken to assess the prevalence of bovine trypanosomosis in some tsetse-infested and tsetse-free areas of Ethiopia. From August 2010 till April 2011, a total of 1524 animals were parasitologically examined and compared by the haematocrit centrifugation technique (Woo test) and polymerase chain reaction (ITS-1 PCR). The ITS-1 PCR was more sensitive and more accurate in species identification than the Woo test. In ITS-1 PCR, an overall trypanosome prevalence of 31.0% was observed that is significantly (P<0.001) higher than in the Woo test (5.3%). Trypanosoma vivax was the predominant taxon (24.9%), followed by T. theileri (6.0%), T. congolense (2.9%) and Trypanozoon (1.6%). Mixed infections were quite common (14% of all infections). The overall prevalence of trypanosome infections in tsetse area (32.4%) was not different from non-tsetse area (30.5%) neither were the prevalences of T. vivax in both areas (respectively 22.6% and 25.7%). With these high prevalences, bovine trypanosomosis continues to hinder animal production and productivity in Ethiopia, both in tsetse-infested and non-infested parts of the country. Attempts to control African trypanosomosis should also pay attention to mechanically transmitted pathogenic trypanosomes and should adopt the most advanced molecular tests for species identification.

Assessment of impacts of tsetse and trypanosomosis control measures on cattle herd composition and performance in southern region, Ethiopia.

This study was conducted to assess the impact of tsetse and trypanosomosis control measures on cattle herd size and composition, herd dynamics, and milk yield in Wolaita and Gamogofa Zones, southern Ethiopia. The study showed that the average number of cattle herd size in tsetse challenged areas was significantly higher than those in tsetse-controlled areas. The number of non-pregnant dry cows, bulls and oxen were significantly higher in tsetse challenged areas than the other two study areas. The rate of cattle addition to and disposal from the herd were significantly higher in tsetse challenged areas. Cows in Southern Tsetse Eradication Project (STEP) and community tsetse controlled areas were able to give 26-27 %, 25-29 % and 17-21 % more daily milk yield at the beginning, middle and end of lactation, respectively, than those in tsetse-challenged areas. In addition, cows in STEP and community tsetse controlled areas had lactation length longer by 1.20 to 1.35 months; age at first calving was shorter by 5.30 to 5.10 months; and calving interval was shorter by 4.20 to 3.20 months than cows in tsetse-challenged area, respectively. Hence, tsetse and trypanosomosis control both by the community and project would play key role in the improvement of cattle productivity.

Sero-prevalence of bovine brucellosis and its risk factors in Jimma zone of Oromia Region, South-western Ethiopia.

A cross sectional sero-prevalence study was conducted on 1,595 cattle in Jimma zone, Ethiopia to investigate the status of bovine brucellosis and identify potential risk factors. Sera samples were analyzed using Rose Bengal Plate Test (RBPT) and Complement Fixation Test (CFT). The overall individual and herd level sero-prevalences were 3.1% (n = 1,595) and 15.0% (n = 227), respectively. The sero-prevalence of bovine brucellosis at individual animal level was significantly higher in non-pregnant (11.18%) than pregnant (2.77%) and lactating (22.35%) than non-lactating animals (2.46%). Moreover, significantly higher sero-prevalence was observed in herds of larger sizes. Individual animal sero-prevalence was also positively associated with the occurrence of abortion (26.98 and 1.54% in those with and without previous history of abortion, respectively). Generally, the sero-prevalence of bovine brucellosis found in Jimma area was not high and the sero-prevalence was closely associated with some of the risk factors considered at individual animal and herd level.

Milk yield and reproductive performance of dairy cattle under smallholder management system in North-eastern Amhara Region, Ethiopia.

This study was conducted in South Wollo Zone of Amhara Regional State, Ethiopia, to assess the milk yield and reproductive performance of indigenous and crossbred cattle under smallholder management conditions. Questionnaire survey was used to collect retrospective data on the performance of dairy cattle in 186 households. Thirty two postpartum cows (16 indigenous and 16 crossbred) were selected purposively and monitored for five months and milk samples were collected twice per week from each cow to determine postpartum ovarian activity using ELISA techniques. Daily milk yields at all stages of lactation were significantly (P < 0.01) higher in crossbred than indigenous cows. Crossbred cows had significantly (P < 0.01) longer lactation length and shorter age at first calving, while calving interval was significantly shorter in cows that gave previous calves during the wet season than the dry season. Only six cows (18.8%) showed heat signs on an average of 136 days postpartum of which one was a crossbred and five were indigenous. While, 11 cows (34.3%) resumed ovarian activity until 150(th) day postpartum among which three were crossbred and eight were indigenous.

Longitudinal observation on reproductive and lactation performances of smallholder crossbred dairy cattle in Fitche, Oromia region, central Ethiopia.

A two-year longitudinal study was conducted to assess the reproductive and lactation performances of smallholder crossbred dairy cattle in Fitche, central Ethiopia. Twenty-four smallholder dairy farms were monitored and data on reproductive, breeding, lactation and management aspects of crossbred dairy cows (n=69) were collected and analysed. Moreover, milk samples were collected and analysed for progesterone. The overall Least-squares means for calving interval (CI), calving to conception interval (CCI) and first observed oestrus after calving were 516, 253 and 141 days, respectively. Suckling significantly prolonged these intervals. The mean first onset of luteal activity (OLA) after calving was 52 days, while 67.4% n=43) had a delayed (>55 days) return to cyclicity after calving. Suckling and parity number significantly (p < 0.05) influenced the OLA. The mean lactation length was 54.4 weeks. The overall daily mean milk yield for the first 43 weeks of lactation after calving was 11.7 L/day. The estimated daily milk yield increased slowly and reached peak (13.8 L/day) around the 11th week post partum and declined gradually and steadily. Suckling status, season of calving and parity number significantly influenced the estimated daily milk yield. In conclusion, the present study indicated that the smallholder crossbred dairy cows in the study area had prolonged intervals to first oestrus after calving, to conception and to the next calving, and thus their reproductive performance is unsatisfactory. Delayed first onset of luteal activity (postpartum anoestrus) contributed to these extended intervals. Further detailed investigation is suggested to examine the effects of other relevant factors on the reproductive and lactation performances of smallholder crossbred dairy cows for appropriate intervention.

Clinical features and hormonal profiles of cloprostenol-induced early abortions in heifers monitored by ultrasonography.

BACKGROUND: The present study describes the clinical features and plasma profiles of bovine pregnancy-associated glycoprotein 1 (bPAG1), the main metabolite of prostaglandin F2alpha (PG metabolite) and progesterone (P4) in heifers in which early abortions were induced. METHODS: Early abortions were induced in four heifers with cloprostenol and monitored by ultrasonography. Blood samples were collected and the plasma were analyzed for bPAG 1, P4 and PG metabolite. RESULTS: The foetal heartbeat rates varied from 170-186 beats per minute for all foetuses up to the date of cloprostenol treatment. Foetal death was confirmed within two days after cloprostenol treatment. Prior to cloprostenol injection, blood plasma concentrations of bPAG1, PG metabolite and P4 varied from 8.4-40.0 ng/mL, 158-275 pmol/L and 20.7-46.9 nmol/L, respectively. After the foetus expelled, the plasma level of bPAG1 began to decrease but the decrease was small and gradual. The estimated half-life of bPAG1 was 1.8-6.6 days. The plasma level of the PG metabolite started to have short lasting peaks (above 300 pmol/L) within three hours after cloprostenol treatment. The plasma concentrations of P4 dropped sharply to less than 4 nmol/L after 24 hours of cloprostenol injection. CONCLUSION: The current findings indicated that after early closprostenol-induced foetal death, the plasma concentration of bPAG1 decreased gradually and showed a tendency of variation with the stages of pregnancy.