Infections with equine herpesviruses (EHVs) are widespread in equine populations worldwide. Whereas both EHV-1 and EHV-4 produce well-documented respiratory syndromes in equids, the contribution of EHV-2 and EHV-5 to disease of the respiratory tract is still enigmatic. This study describes the detection and genetic characterization of EHVs from equids with and without clinical respiratory disease. Virus-specific PCRs were used to detect EHV-1, -2, -4 and -5. From the total of 160 equids with respiratory disease, EHV-5 was detected at the highest prevalence (23.1%), followed by EHV-2 (20.0%), EHV-4 (8.1%) and EHV-1 (7.5%). Concurrent infections with EHV-2 and EHV-5 were recorded from nine (5.2%) diseased horses. Of the total of 111 clinically healthy equids, EHV-1 and EHV-4 were never detected whereas EHV-2 and EHV-5 were found in 8 (7.2%) and 18 (16.2%) horses, respectively. A significantly higher proportion of EHV-2-infected equids was observed in the respiratory disease group (32/160, 20.0%; P = 0.005) compared to those without disease (8/111; 7.2%). EHV-2-positive equids were three times more likely to display clinical signs of respiratory disease than EHV-2-negative equids (OR 3.22, 95% CI: 1.42-7.28). For EHV-5, the observed difference was not statistically significant (P = 0.166). The phylogenetic analysis of the gB gene revealed that the Ethiopian EHV-2 and EHV-5 strains had a remarkable genetic diversity, with a nucleotide sequence identity among each other that ranged from 94.0 to 99.4% and 95.1 to 100%, respectively. Moreover, the nucleotide sequence identity of EHV-2 and EHV-5 with isolates from other countries acquired from GenBank ranged from 92.9 to 99.1% and 95.1 to 99.5%, respectively. Our results suggest that besides EHV-1 and EHV-4, EHV-2 is likely to be an important contributor either to induce or predispose equids to respiratory disease. However, more work is needed to better understand the contribution of EHV-2 in the establishment of respiratory disease.

BackgroundMannheimia haemolytica has been recognized as the principal cause of pneumonic pasteurellosis in sheep and goats. It is one of the important diseases of small ruminants in Ethiopia. While annual vaccination using a monovalent vaccine (inactivated Pasteurella multocida biotype A) is common, respiratory diseases are still reported in various parts of Ethiopia. This suggests the need for further investigation into the species and strains responsible for the disease, which is vital information for development of a multivalent vaccine. The objective of the current study was to isolate M. heamolytica associated with pneumonic cases of sheep in selected areas of Central Ethiopia, determine its role and the strains/genotypes of the bacterium circulating in the study area.ResultsBacteriological analysis of nasal swab samples collected from a total of 76 pneumonic cases of sheep showed that M. haemolytica was isolated from 26 of them while B.trehalosi from two cases. Further molecular analyses of the isolates using M. haemolytica species-specific and M.haemolytica serotype-1 antigen specific PCR assays revealed, 26 of the isolates were identified as M. haemolytica of which 21 of them were M. haemolytica serotype-1. Both M. haemolytica and B.trehalosi isolates were not detected in a PCR assay targeting capsular biosynthesis gene (capA) of P.multocida despite the non-specific products observed in M. haemolytica isolates. Phylogenetic analysis of M. haemolytica isolates included in this study in comparison with the reference strains with respect to PHSSA and Rpt2 genes revealed that the Ethiopian M. haemolytica isolates constituted three distinct genotypes consistent with site of origin.ConclusionThe study indicated that M.haemolytica is commonly associated with cases of pneumonia in sheep in the study areas of central Ethiopia although the remaining other pathogens responsible for majority of the cases are yet to be determined. Molecular characterization revealed the existence of three genotypes of M. haemolytica circulating in the study areas consistent to the site of isolation. The findings suggest further extensive work to determine all pathogens associated with sheep pneumonia and the strain distribution of M. heamolytica to understand its molecular epidemiology at national level and design cost effective prevention and control methods.

A cross sectional study was conducted on dairy items in Addis Ababa from October 2010 to March 2011 to determine prevalence and antimicrobial resistance profile of Salmonella. A total of 384 dairy items, 96 of each item (cheese, milk, butter and yogurt) was sampled. The overall prevalence of Salmonella was found to be 1.6% (6 of 384). Prevalence of 3.1, 1.04, 2.1, and 0% was observed from cheese, butter, milk and yogurt, respectively. However, there was no statistically significant difference (P > 0.05) in the prevalence of Salmonella among the different sample types. Isolates were tested for the effects of eight antimicrobials by disk diffusion technique; all isolates were resistant to one or more of the tested antimicrobials. Of all isolates, 50% were multiple antimicrobial resistant. 83.3, 50, 16.7, and 16.7% of isolates were resistant to tetracycline, ampicillin, amoxicillin, and chloramphenicol, respectively. However, all the isolates were susceptible to gentamycin, ceftriaxone, ciprofloxacin, and sulfamethoxazole. From this pilot study, we concluded that dairy products are a potential source of Salmonella infection with antimicrobial resistance. Furthermore, hygienic management of dairy products and prudent use of antimicrobials are also suggested.

Pasteurella multocida can infect a multitude of wild and domesticated animals, with infections in cattle resulting in hemorrhagic septicemia (HS) or contributing to bovine respiratory disease (BRD) complex. Current cattle vaccines against P. multocida consist of inactivated bacteria, which only offer limited and serogroup specific protection. Here, we describe a newly identified surface lipoprotein, PmSLP, that is present in nearly all annotated P. multocida strains isolated from cattle. Bovine associated variants span three of the four identified phylogenetic clusters, with PmSLP-1 and PmSLP-2 being restricted to BRD associated isolates and PmSLP-3 being restricted to isolates associated with HS. Recombinantly expressed, soluble PmSLP-1 (BRD-PmSLP) and PmSLP-3 (HS-PmSLP) vaccines were both able to provide full protection in a mouse sepsis model against the matched P. multocida strain, however no cross-protection and minimal serum IgG cross-reactivity was identified. Full protection against both challenge strains was achieved with a bivalent vaccine containing both BRD-PmSLP and HS-PmSLP, with serum IgG from immunized mice being highly reactive to both variants. Year-long stability studies with lyophilized antigen stored under various temperatures show no appreciable difference in biophysical properties or loss of efficacy in the mouse challenge model. PmSLP-1 and PmSLP-3 vaccines were each evaluated for immunogenicity in two independent cattle trials involving animals of different age ranges and breeds. In all four trials, vaccination with PmSLP resulted in an increase in antigen specific serum IgG over baseline. In a blinded cattle challenge study with a recently isolated HS strain, the matched HS-PmSLP vaccine showed strong efficacy (75–87.5% survival compared to 0% in the control group). Together, these data suggest that cattle vaccines composed of PmSLP antigens can be a practical and effective solution for preventing HS and BRD related P. multocida infections.

This experimental study was done on a total of 40 male lambs with the objectives of developing experimental vaccines from Mannheimia haemolytica serotypes A2 and A7 that express iron regulated outer membrane protein and in vivo evaluation of their efficacy. Lambs were categorized in to four experimental groups and vaccinated with 1 ml of vaccine containing 5 × 10 8 CFU/ml. Group 1 was vaccinated with M. haemolytica A2, group 2 with A7, group 3 with serotype A2 and A7 combination, and group 4 received saline as control. They were challenged intratracheally by the respective homologous serotype after 35 days of vaccination. Post challenge clinical investigation showed that significant higher rate of morbidity was seen in control group which was demonstrated by raised rectal temperature (by 0.5-1°C) and respiratory signs. From the total of 26 lambs challenged with live M. haemolytica A2 and A7, 6 (23.1%) and 4 (15.3%) lambs were found dead and sick, respectively. Higher mortality and morbidity were observed in unvaccinated control group; however, lesser was recorded in combined vaccinated group. Lung lesions of variable severity were observed in 13 (50.0%) lambs following challenge. From vaccinated groups, 5 (27.8%) lambs were found to have a +1 lung lesion score. All of the lambs in unvaccinated control group had scores between +2 and +3. There was a statistically significant difference (p < 0.05) between control and vaccinated groups, while no statistically significant difference (p > 0.05) was seen among vaccinated groups concerning lung lesion scores. Furthermore, the respective serotypes of M. haemolytica were successfully re-isolated from pneumonic lungs at a mean titre range of 10 2.2 -10 8.1 CFU/g. In conclusion, lambs which received combined vaccine confer relatively good protective efficacy than M. haemolytica A2 or A7 vaccinated groups. Therefore, further study should be done on evaluation of antibody titer at different time points.