Contagious caprine pleuropneumonia (CCPP) is a fatal disease of goats occurring in many countries ofAfrica and Asia where the total goat population is more than 500 million. Vaccination is the most cost effective technique in the control of CCPP than any other control measures. In National Veterinary Institute (NVI) inactivated mycoplasma protein based vaccine obtained by centrifugation has been in use since many years. This study focuses on evaluating the safety and immunogenicity of inactivated whole culture CCPP vaccine currently developed in the NVI. Twenty six Mycoplasma capricolum subspecies capripneumoniae (Mccp) antibody free goats were used to evaluate the safety and immunogenicity of inactivated whole culture CCPP trial vaccine. The trial vaccine was prepared from culture of Mccp vaccinal seed grown in Mycoplasma specific hayflick media using spinner bottle. The protein content for one milliliter of whole culture was checked and found to be more than the minimum recommended dose (0.15 mg per dose). The culture was inactivated by 37% formalin at proportion of 0.5% of whole culture and adjuvanted by saponin at final concentration of 0.3%. The experimental animals were distributed into four groups: The group A consist of five goats for safety and all the other groups consists of seven animals each with group B for trial vaccine, group C for positive control and group D for negative control for immunogenicity trials. The goats were observed for two months for safety and immunogenicity evaluation during which serum samples were collected for immunogenicity and tested by using competitive Enzyme Linked immunosorbent Assay (cELISA) test. The results indicated that out of 7 goats vaccinated with trial vaccine, the mean sero-positivity was 60.71% while 7 goats vaccinated with the positive control showed mean sero-positivity of 58.86%. The analysis showed no significant difference between mean sero positivity of trial vaccine and positive control (P>0.05) as indicated by sero-conversion. The mean percent inhibition (PI) of trial inactivated whole culture CCPP vaccine vaccinated goats was 61.52% while the mean PI for positive control vaccine vaccinated group was 51.86%. In contrast the non-vaccinated controls showed mean PI of 40.65% which is significantly less than percent inhibition of the vaccinated groups (p=0.000). The body temperature and clinical observation of safety tested animals and other immunogenicity tested goats showed absence of any abnormality after vaccination both in vaccinated and controls. This study which was novel in its nature concluded that the trial inactivated whole culture ccpp vaccine is equally immunogenic as that vaccine already in use, the non-whole culture concentrated CCPP vaccine, and could be used for mass vaccination after conducting field immunogenicity trial.
Respiratory diseases caused by Mannheimia haemolytica (M. haemolytica) and Pasteurella multocida (P. multocida) have been known to result in a considerable loss due to mortality and reduced production. This study aimed at isolation and identification of M. haemolytica and P. multocida associated with pneumonic pasteurellosis in sheep and goats using bacteriological and molecular techniques. Identification of serotypes of M. haemolytica and P. multocida was done using indirect haemagglutination test. The in vitro antimicrobial sensitivity profiles of the M. haemolytica were tested using standard disk diffusion method. A total of 52 and 78 nasal swabs were collected from pneumonic cases for bacterial isolation and identification in Borana and Arsi zone, respectively. Four hundred sera samples were collected for identification of serotypes. The results showed that 17 of 52 (32.69%; 95% CI 20.33, 47.11) nasal swabs collected from pneumonic animals in Borana yielded positive results for Pasteurella/Mannheimia species, 13 (25.00%; 95% CI 14.03, 38.95) of which were M. haemolytica. None of the samples yielded P. multocida. Twenty-three of 78 (29.49%; 95% CI 19.69, 40.89) nasal swabs collected at Arsi from pneumonic animals yielded positive results for M. haemolytica (17) and P. multocida (6). Secondary biochemical characterization revealed that 14 of the 17 isolates conform to M. haemolytica whereas none of the 6 isolates suspected to be P. mutocida were confirmed. Eleven (84.62%) isolates from Borana and 4 (28.57%) from Arsi were confirmed to be M. haemolytica using PCR targeting the Rpt2 genes. Assay for M. haemolytica serotype A1 revealed all belong to A1. None of the isolates with cultural and morphological features of P. multocida gave positive results by molecular assay. Serological assay identified three serotypes of M. haemolytica namely A1, A2 and A7 almost in all of the samples whereas P. multocida serotype A was detected in 78.75% of the samples. The M. haemolytica isolates tested for susceptibility to antibiotics showed resistance against Bacitracin (83.33%) and Penicillin (50.00%) while they were found susceptible to Gentamycin (100%), Chloramphenicol (100%) and Sulfamethoxazole (100%) and Tetracycline (83.33%). In conclusion, the results of the present study revealed the association of M. haemolytica with pneumonic pasteurellosis in sheep and goats and can be of use in vaccine development in Ethiopia. Nevertheless, further investigations and continuous monitoring of antimicrobial resistance and appropriate selection and prudent use of antimicrobials in livestock sector are required.
Respiratory diseases caused by M. haemolytica and P. multocida have been known to result in a considerable loss due to mortality and reduced production. This study aimed at isolation and identification of M. haemolytica and P. multocida associated with pneumonic pasteurellosis in sheep and goats using bacteriological and molecular techniques. Identification of serotypes of M. haemolytica and P. multocida was done using indirect haemagglutination test (IHAT). The in vitro antimicrobial sensitivity profiles of the M. haemolytica were tested using standard disk diffusion method. A total of 52 and 78 nasal swabs were collected from pneumonic cases for bacterial isolation and identification in Borana and Arsi zone, respectively. Four hundred sera samples were collected for identification of serotypes. The results showed that 17 of 52 (32.69%; 95% CI: 20.33, 47.11) nasal swabs collected from pneumonic animals in Borana yielded positive results for Pasteurella / Mannheimia species, 13 (25.00%; 95% CI: 14.03, 38.95) of which were M. haemolytica. None of the samples yielded P. multocida. Twenty-three of 78 (29.49%; 95% CI: 19.69, 40.89) nasal swabs collected from pneumonic animals yielded positive results for M. haemolytica (17) and P. multocida (6). Secondary biochemical characterization revealed that 14 of the 17 isolates conform to M. haemolytica whereas none of the 6 isolates suspected to be P. mutocida were confirmed. Eleven (84.62%) isolates from Borana and 4 (28.57%) from Arsi were confirmed to be M. haemolytica using PCR targeting the Rpt2 genes. None of the isolates with cultural and morphological features of P. multocida gave positive results by molecular assay. Serological assay identified three serotypes of M. haemolytica namely A1, A2 and A7 almost in all of the samples whereas P. multocida serotype A was detected in 78.75% of the samples. The M. haemolytica isolates tested for susceptibility to antibiotics showed resistance against Bacitracin (83.33%) and Penicillin (50.00%) while were found susceptible to Gentamycin, Chloramphenicol and Sulfamethoxazole (100%) and Tetracycline (83.33%). In conclusion, the results of the present study revealed the association of M. haemolytica with pneumonic pasteurellosis in sheep and goats and can be of use in vaccine development in Ethiopia. Nevertheless, further investigations and continuous monitoring of antimicrobial resistance and appropriate selection and prudent use of antimicrobials in livestock sector are required.
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