Improving Quality Control of Contagious Caprine Pleuropneumonia Vaccine with Tandem Mass Spectrometry.

Vaccines to protect livestock against contagious caprine pleuropneumonia (CCPP) consist of inactivated, adjuvanted antigens. Quality control of these vaccines is challenging as total protein quantification provides no indication of protein identity or purity, and culture is not an option. Here, a tandem mass spectrometry approach is used to identify the mycoplasma antigen contained in reference samples and in commercial CCPP vaccines. By the same approach, the relative amounts of mycoplasma antigen and residual proteins originating from the production medium are determined. Mass spectrometry allows easy and rapid identification of the peptides present in the vaccine samples. Alongside the most probable mycoplasma species effectively present in the vaccines, a very high proportion of peptides from medium constituents are detected in the commercial vaccines tested.

Prevalence of contagious caprine pleuro-pneumonia in pastoral flocks of goats in the Rift Valley region of Kenya.

A cross-sectional survey was conducted between the months of March 2014 and March 2015 to determine the prevalence of contagious caprine pleuropneumonia in goat populations in pastoral flocks in three sub-counties of the Rift Valley region. A total of 432 serum samples were collected from goats from 54 flocks and tested for the presence of antibodies against mycoplasma capricolum subspecies capripneumoniae (mccp) using monoclonal antibody-based competitive enzyme-linked immuno-sorbent assay. Sero-prevalence recorded for Turkana West was 63.9%, Kajiado Central was 48.6%, while Pokot East was 29.2% which was statistically significant (χ2 = 34.997; P = 0.000) in the study sites. The results of this study confirmed that CCPP is widespread and endemic in the pastoral production systems studied in the Rift Valley region. The results confirmed that regions sharing international boundaries are at a higher risk of CCPP hence the need for a unified cross-border approach to disease control measures in the border areas.

Antibody response to Mycoplasma capricolum subsp. capripneumoniae bacterium in small holder dairy goats in Tanzania.

Contagious caprine pleuropneumonia (CCPP), an Office International des Epizooties listed disease, can cause significant levels of morbidity and mortality in goats in Africa, the Middle East, and Western Asia. A cross-sectional study was conducted to determine the seroprevalence of CCPP, in two contrasting administrative districts of northern Tanzania, namely, Babati and Arumeru. A total of 337 serum samples were collected from January to July 2010, from apparently clinically healthy unvaccinated dairy goats breeds of different ages and sexes. Samples were subjected to monoclonal antibody-based competitive enzyme-linked immunosorbent assay for the specific measurement of antibodies to Mycoplasma capricolum subsp. capripneumoniae bacterium. The overall animal flock and village-level seroprevalence of CCPP was found to be 3.3 % [95 % confidence interval (CI) = 1.6-5.8], 9.6 % (95 % CI = 4.7-16.9), and 31.5 % (95 % CI = 15.6-56.5), respectively. There was no evidence (p = 0.659) of differences in seroprevalence between samples from the two administrative locations. Despite the fact that there was no significant difference (p > 0.05) in seropositivity between sex and age and between breeds sampled, there was a significant difference (p < 0.05) in seropositivity between the different physiological status groups examined with the non-lactating (dry) category of goats showing a higher seroprevalence. The findings of this survey revealed evidence of dairy goat exposure to M. capricolum subsp. capripneumoniae, although at a low prevalence. It is therefore advisable to include CCPP serology in the seromonitoring program so as to give a better indication of flock immunity. This should lead to the establishment of appropriate CCPP control measures in smallholder dairy goat flocks, which are increasingly being recognized for their value as a vital source of livelihood for resource poor livestock keepers in Tanzania.

Cross-sectional study on Contagious Caprine Pleuro Pneumonia in selected districts of sedentary and pastoral production systems in Southern Ethiopia.

A study to estimate the seroprevalence of Contagious Caprine Pluropneumonia (CCPP) in southern Ethiopia was conducted from November 2005 to June 2006. Two districts from sedentary (Arbaminch and Boreda) and pastoral (Hammar and Bena-Tsemay) production systems were included in the study. Sera samples were collected from 913 goats (234 from sedentary and 679 from pastoral) to check for CCPP serostatus. The animals were sampled from 155 flocks (44 pastoral and 111 sedentary). Five clinically suspected CCPP cases were also sacrificed and attempt was made to isolate Mycoplasma capricolum capripneumoniae (MccP) from lung tissue, nasal swab and plural exudates. Sera samples were tested for the presence of CCPP antibodies using CFT. The overall seroprevalence recorded in the study was 18.61%. The corresponding seroprevalences for sedentary and pastoral production systems were 27.78% and 15.46% respectively. Regarding districts, the prevalence in Hammar was 15.63% while that of Bena-Tsemay 15.29%. In Arbaminch and Boreda the percent of seroreactors were 23.01 and 32.23% respectively. Out of 44 pastoral and 111 sedentary flocks, 50.45% of pastoral and 65.91% of sedentary flocks had at least one seroreactor goat per flock respectively. Both in the univariable and multivariable logistic regression analysis, seropositivity was found to have strong association with sedentary production system (P < 0.05, OR = 2.24) and adult age (P < 0.05, OR = 1.77). In microbiological study, two broth cultures from thoracic fluid and two broth cultures from lung tissue samples were found to be positive for Mycoplasma capricolum capripneumoniae (MccP). In conclusion, both the serological study and bacteriological isolation confirmed the disease CCPP being an important disease that demands serious attention in both production systems.

Interleukin-17 mediates lung injury by promoting neutrophil accumulation during the development of contagious caprine pleuropneumonia.

Contagious caprine pleuropneumonia (CCPP) is a highly contagious infectious disease of goats caused by Mycoplasma capricolum subspecies capripneumoniae (Mccp). CCPP outbreaks usually result in high morbidity and mortality of the affected goats, making this disease a major cause of economic losses to goat producers globally. However, the pathogenesis of CCPP remains unclear. Here, we show that IL-17-driven neutrophil accumulation is involved in the lung damage in CCPP goats. During CCPP development, intense inflammatory infiltrates could be observed in the injured lungs. Specifically, neutrophils were observed to be present within the alveoli. Increased IL-17 release drove the excessive influx of neutrophils into the lung, as IL-17 effectively stimulated the production of neutrophil chemoattractants from lung epithelial cells following Mccp infection. Our data highlight a critical role of IL-17-driven neutrophil accumulation in the pathogenesis of CCPP and suggest that IL-17 may potentially be a useful immunotherapeutic target for the treatment of CCPP.

Characterisation of Mycoplasma capricolum P60 surface lipoprotein and its evaluation in a recombinant ELISA.

This paper reports the identification and characterisation of a 60kDa surface protein antigen (P60) of Mycoplasma capricolum subspecies capricolum (Mcc), and describes its diagnostic application. Genomic localization and presence in P60 of conserved functional domains suggested a structural and functional relationship with the immunodominant antigen P48 of Mycoplasma agalactiae, a basic membrane protein. A rP60-ELISA was developed, and it resulted in a high specificity for Mcc infections after evaluation with 125 goat sera. The comparison with an existent ELISA based on whole Mcc cell lysates showed that the two assays have comparable sensitivities, but the rP60-ELISA has the significant advantage of a greater specificity. Results indicate that P60 is a potential marker of Mcc infection, especially useful in areas where the presence of M. capricolum subspecies capripenumoniae is also reported.

Experimental contagious caprine pleuropneumonia: a long term study on the course of infection and pathology in a flock of goats infected with Mycoplasma capricolum subsp. capripneumoniae.

Contagious caprine pleuropneumonia (CCPP) is a major threat to goat farming in parts of Africa and Asia. It classically causes acute high morbidity and mortality early in infection, but little is known of its long term epizootiology and course. In this study, 10 goats were inoculated with Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae) and then mixed with 15 goats for contact transmission. The disease course was monitored in each goat for 56-105 days, whereafter the goats were killed and necropsied. Varying features signifying infection occurred in altogether 17 goats (7 inoculated, 10 in-contact). Clinical signs were severe in 8 goats but no fatalities occurred. Only 6 goats had serum antibody titres against M. capripneumoniae in ELISA. Fourteen goats (5 inoculated, 9 in-contact) had chronic pleuropulmonary lesions compatible with CCPP at necropsy and 7 of those showed M. capripneumoniae antigen in the lung by immunohistochemistry. Neither cultivation nor PCR tests were positive for the agent in any goat. The results indicate that the clinical course of CCPP in a flock may be comparatively mild, M. capripneumoniae-associated lung lesions may be present at a late stage of infection, and chronic infection may occur without a significant serological response.

Identification of novel immunogenic proteins in Mycoplasma capricolum subsp. Capripneumoniae strain M1601.

The purpose of this study was to obtain immunogenic proteins and potential proteins of interest that were isolated from Mycoplasma capricolum subsp. capripneumoniae (Mccp) by MALDI-TOF mass spectrometry. One-dimensional SDS-PAGE and two-dimensional gel electrophoresis of whole cell preparation were conducted, and membrane proteome maps were prepared by immunoblotting. One-dimensional SDS-PAGE identified three immunogenic proteins with molecular masses in the range 29-97.2 kDa, two of which were in the membrane protein fraction. After two-dimensional gel electrophoresis, 20 highly immunogenic proteins were identified in the whole cell protein preparation while 9 immunogenic proteins were identified in the membrane protein fraction. This indicated that membrane proteins were the principle immunogenic proteins in Mccp. These proteins may have potential for the development of improved diagnostic tests and possible vaccines.

A study on contagious caprine pleuropneumonia (CCPP) in goats at an export oriented abattoir, Debrezeit, Ethiopia.

300 goat serum samples from an export-oriented abattoir were tested for contagious caprine pleuropneumonia antibodies by the complement fixation test. The disease prevalence was 31% with no significant differences (P > 0.05) between the regions "Borena", "Bale", "Afar" and "Jinka" or the age of the goats (P > 0.05). Gross pathology and histopathology of the lung primary lesions were indicative of pleuropneumonia caused by Mycoplasma capricolum subsp. capripneumoniae.