Waseem Shahzad

Global suppression of mitogen-activated ovine peripheral blood mononuclear cells by surface protein activity from Mycoplasma ovipneumoniae.

Mycoplasma ovipneumoniae is associated with chronic non-progressive pneumonia of sheep and goats. As with many other mycoplasmas involved in animal diseases, protective immune responses have not been achieved with vaccines, even though antibody responses can be obtained. This study focuses on characterizing the interaction of M. ovipneumoniae with ovine PBMC using carboxy-fluorescein-succinimidyl-ester (CFSE) loading and flow cytometry to measure lymphoid cell division. M. ovipneumoniae induced a strong in vitro polyclonal suppression of CD4(+), CD8(+), and B blood lymphocyte subsets. The suppressive activity could be destroyed by heating to 60 degrees C, and partially impaired by formalin and binary ethyleneimine treatment that abolished its viability. The activity resided on the surface-exposed membrane protein fraction of the mycoplasma, since mild trypsin treatment not affecting viability was shown to reduce suppressive activity. Trypsin-treated mycoplasma regained suppressive activity once the mycoplasma was allowed to re-synthesize its surface proteins. Implications for the design of vaccines against M. ovipneumoniae are discussed.

Prevalence and molecular diagnosis of Trypanosoma evansi in Nili-Ravi buffalo (Bubalus bubalis) in different districts of Punjab (Pakistan)

The prevalence of Trypanosoma evansi was investigated in 1,250 Nili-Ravi buffaloes of mixed age and sex by polymerase chain reaction (PCR) for the first time in Pakistan. DNA of the trypanosomes was isolated with TRIREAGENT®. The assay was employed using primers ESAG 6/7, specific for a 237-bp fragment from T. evansi genomic DNA. The samples were screened for the presence of T. evansi also by stained thin smear. Forty-four (3.5%) samples were positive by microscopy, while 97 (7.7%) samples were identified by PCR, indicating the high sensitivity of PCR for surveying the disease in epidemiological studies.