Detection of filarial parasites and Wolbachia endosymbiotic bacteria in mosquitoes by PCR

Wuchereria Bancroft; the principal etiologic agent of lymphatic filariasis is mosquito dependant in the biological transmission. Dirofilariasis is essentially a disease of canines which can also be trans-mitted to humans by culicine mosquitoes. Wolbachia are Rickettsia-like, obligatory intracellular bacteria that infect the reproductive and somatic tissues of some arthropods and nematodes. Our study aimed to identify the possible association between filarial parasites and Wolbachia by single and multiplex PCR. 1600 female mosquitoes collected from: four localities in Assiut Governorate, Egypt were microscopically identified and divided into 64 mosquito pools according to their genera and collection site. Single PCR was firstly conducted on mosquitoes followed by multiplex PCR for simultaneous detection of the three filarial parasites [Wucheraria bancrofti, Dirofilaria immitis, and Dirofilaria repens] and Wolbachia in mosquitoes. The results indicated that: Out of 64 mosquito pools, 8 pools were positive for Wuchererio bancrofti with estimated rate of infection [ERI 0.53%], 3 for each of Dirofilaria immitis and Dirofilaria repens [ERI 0.19%] and 6 pools were positive for Wolbachia [ERI 0.39%], five of them with filarial parasites [W. Bancrofti, D. immitis and D repens] [83.3%], with a significant P. value [< 0.05]. We concluded that there was a strong association between the presence of Wolbachia and filarial parasites that should be considered during the treatment of patients with filarial diseases by the use of specific anti-Wolbachia antibiotic in addition to the usual anti-filarial drugs

Immunodiagnosis of capillaria philippinensis by western blot using coproantigen and egg antigen

Capillaria philippinensis is a zoonotic nematode that causes severe diarrhoeal syndromes in humans. C. philippinensis coproantigen and egg antigen were fractionaled using SDS-PAGE. Several protein bands were detected in both antigens. Protein profile of coproantigen of C. philippinensis infected patient included wide range of bands ranging from 10 - 180 kD. Protein profile of egg antigen revealed few bands detected at 10, 34- 36.5, and 148 kD. Using Western blot, a variety of protein bands were recognized by the infected patient serum. When tested against coproantigen, it detected a wide range of antigenic proteins at different molecular weights [10, 14, 34-36.5, 48, 54, 62, 66, 86, 94, and 148 kD respectively]. None of these bands were detected in normal human feces. When tested against egg antigen, the infected patient serum detected only two faint antigenic bands at 62 and 66 kD. It is concluded that coproantigen is more convenient than egg antigen in diagnosis of capillariasis by WB