Finding Wolbachia in Filarial larvae and Culicidae Mosquitoes in Upper Egypt Governorate

Wolbachia is an obligatory intracellular endosymbiotic bacterium, present in over 20% of all insects altering insect reproductive capabilities and in a wide range of filarial worms which is essential for worm survival and reproduction. In Egypt, no available data were found about Wolbachia searching for it in either mosquitoes or filarial worms. Thus, we aimed to identify the possible concurrent presence of Wolbachia within different mosquitoes and filarial parasites, in Assiut Governorate, Egypt using multiplex PCR. Initially, 6 pools were detected positive for Wolbachia by single PCR. The simultaneous detection of Wolbachia and filarial parasites (Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens) by multiplex PCR was spotted in 5 out of 6 pools, with an overall estimated rate of infection (ERI) of 0.24%. Unexpectedly, the highest ERI (0.53%) was for Anopheles pharoensis with related Wolbachia and W. bancrofti, followed by Aedes (0.42%) and Culex (0.26%). We also observed that Wolbachia altered Culex spp. as a primary vector for W. bancrofti to be replaced by Anopheles sp. Wolbachia within filaria-infected mosquitoes in our locality gives a hope to use bacteria as a new control trend simultaneously targeting the vector and filarial parasites.

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

Comparative study between non lethal and lethal strains of Plasmodium yoelii with reference to its immunological aspect

Innate immunity has an important role in the protection against malaria. To clarify the effect on non lethal and lethal strain of Plasmodium yoelii, comparison between two groups of C57BL/6 mice infected with 10[4] parasitized RBCs was performed. Liver and spleen mononuclear cells were isolated and analyzed by flow cytometry. The parasite appeared in blood on day 3 in both strains, with non lethal infection parasitemia reached a peak of 60% on day 14 and mice completely recovered, while in lethal infection parasitemia was 80% on day 7 and mice succumbed to death. In non lethal strain, mice became anemic and the hematocrit percentage returned to its normal value during recovery, while in the lethal strain mice were severely anemic before death. The major expanding cells were found to be TCR Intermediate [TCR[int]] cells, mainly NK1.1 subset, these TCR[int] cells were distinguished from conventional T cells of thymic origin. CD4[-] and CD8[-] cells increased in both strains. During malarial infection, the population of conventional T cells did not increase and usually associated with thymic atrophy. The present results showed that TCR[int] cells were intimately associated with the protection against malarial infection in both non lethal and lethal strains but the mice died in lethal infection due to the massive destruction of red blood cells leading to fatal anemia

Determination of antigenic patterns of three taenia extracts and their use for differential diagnosis of taeniasis by immunoblot

Immunoserological differential diagnosis of Taenia spp. is the major contribution of the present work. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis [SDS-PAGE] was used to analyze the protein components of three Taenia extracts prepared from Cysticercus bovis, Taenia saginata and Taenia taeniaeformis. Antigenic components of the three taeniid antigens revealed by [SDS-PAGE] were: [I] Cysticercus bovis cystic fluid antigen: 180, 116, 90, 66, 45, 26, 17 and 10 kD. [II] Taenia saginata excretory secretory antigen [ES]: 198, 180, 146, 116, 55, 45, 35 and 26 kD. [III] Taenia taeniaeformis whole crude antigen 170, 140, 120, 116, 66, 55, 45, 35, 26, 17 and 10 kD. The characterized extracts were used as antigens in Western Blot [WB] technique. Sera from immunized rabbits with Cysticercus bovis fluid antigens were used in detecting a variety of immunogenic bands: [I] Cysticercus bovis: 160, 116, 86, 52, 45, 38 and 29 kD. [II] Taenia saginata: 86, 68 and 10 kD. [III] Taenia taeniaeformis: 170, 116, 86, 68, 52, 45 and 29 kD. The present results suggested that the diagnostic bands by Western Blotare 160 and 38 kD for Cysticercus bovis, 10 kD for Taenia saginata [ES] antigen and 170 kD for Taenia taeniaeformis

Oxyurid nematodes detected by cotonoscopy in patients with unexplalnea abdominal pain

Pinworms are one of the common helminthic infection that generally live in the gastrointestinal tract causing appendicitis and leading to unexplained abdominal pain. Species of the genus Syphacia [rodent pinworm] are cosmopolitan and they also infect humans. To diagnose the cause of unexplained abdominal pain in patients with mild eosinophilia by colonoscopy. to detect the relevance of Oxiyurid nematodes as a cause of this unexplained abdominal pain, and to identify and describe the extracted piliworms using light and scanning electron microscopy [SEM]. The study was performed on 200 inpatients of different age groups ranging from 3-60 years over a period of one year in the Tropical Medicine and Gastroenterology Department, Assiut University Hospital. Laboratory investigations were done for each case, including complete blood picture, liver function tests, stool examination for helminthes and protozoa. and perianal swab for patients suffering from perianal itch. Colonoscopy was performed for all cases not responding to antispasmodics. Detected worms were picked up by biopsy forceps and sent to the Parasitology Department, Faculty of Medicine. Assuit University and examined using light and SEM. Out of 200 patients, 25[12.5%] were diagnosed as pinworm infection of the genus Syphacia except in 5 children who had mixed infection with E. vermicularis. Laboratory findings were mild cosinophilia [6-8%] and neutrophulia with moderate shift to the left in one patient with recto-sigmoid nodule and negative stool examination. perianal swab of patients presenting with perianal itch was positive for E. vermicularis eggs. Light microscopic examination illustrated the presence of three different species of Oxyurida E. vermicularis. Syphacia muris and .Syphacia specific. SF.M studies showed that Svphacia spp. were classified into two groups according to morphological differences, and allowed for the reporting of additional morphological and taxouomical features. Syphhacia is considered as a cause of unexplained chronic abdominal pain and E. vermicularis is not the only human pinworm in Egypt. Further studies using SEM are needed to detect new characters that may help in differentiating Syphacia spp. from different hosts