Ahmed Elbehairy

Stray dogs are considered as sentinels in the epidemiology of Toxoplasma gondii because they are carnivores and eat variety of foods, including garbage. In the present study, tissues and sera of 51 stray dogs (Canis familiaris) from Giza, Egypt were examined for T. gondii infection. Sera were examined for antibodies to T. gondii by the modified agglutination test (MAT); 50 of 51 (98%) were seropositive with titers of 20 in four, 40 in four, 80 in one, 100 in eight, 200 in 17, 400 in 11, 800 or higher in five. Hearts of 43 seropositive dogs were bioassayed in mice. Viable T. gondii was isolated from 22 dogs; these isolates were designated TgDogEg1 to TgDogEg22. DNA isolated from cell culture derived tachyzoites of 22 isolates was genotyped using 10 PCR-restriction fragment length polymorphism markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico). The results revealed three genotypes and one mixed infection. The three genotypes are ToxoDB PCR-RFLP #2
(type III, four isolates), #3 (type II variant, 11 isolates), #20 (six isolates), 1 mixed infection. These results revealed the dominance of clonal type II, III and ToxoDB #20 lineages of T. gondii in stray dogs from Giza, Egypt

Leishmaniasis is an insect-transmitted parasitic disease with a worldwide distribution. Leishmania spp. infections cause a broad spectrum of clinical signs, ranging from skin lesions to fatal visceral disease. Dogs are a major reservoir host for visceral leishmaniasis in humans. While the disease is endemic in the Middle East and North Africa, little is known concerning canine Leishmania spp. infections in Egypt. Accordingly, blood samples were collected from 50 stray dogs in Giza, Egypt. Canine sera were tested for antibodies to visceralizing Leishmania spp. by commercial immunochromatographic strip assays based on recombinant antigen K39. Antibodies to Leishmania spp. were found in 5 of 50 (10%) of dogs tested from Egypt. Results from this study indicate that stray dogs are exposed to visceralizing Leishmania species in Egypt.

The present study aimed to establish a fluorescencebased polymerase chain reaction-linked single-strand
conformation polymorphism (F-PCR-SSCP) assay for the identification of Fasciola spp. Based on the sequences
of the second internal transcribed spacer (ITS-2) of the nuclear ribosomal DNA, we designed a set of genusspecific primers for the amplification of Fasciola ITS-2, with an estimated size of 140 bp. These primers were labelled by fluorescence dyes, and the PCR products were analyzed by capillary electrophoresis under nondenaturing conditions (F-PCR-SSCP). Capillary electrophoresis analysis of the fluorescence-labelled DNA fragments displayed three different peak profiles that allowed the accurate identification of Fasciola species: one single peak specific for either Fasciola hepatica or Fasciola gigantica and a doublet peak corresponding to the
“intermediate” Fasciola. Validation of our novel method was performed using Fasciola specimens from different
host animals from China, Spain, Nigeria, and Egypt. This F-PCR-SSCP assay provides a rapid, simple, and robust tool for the identification and differentiation between Fasciola spp.

Real time quantitative PCR (qPCR) is one of the key technologies of the post-genome era, with clear advantages compared to normal end-point PCR. In this paper, we report the first qPCR-based assay for the identification of Fasciola spp. Based on sequences of the second internal transcribed spacers (ITS-2) of the ribosomal rRNA gene, we used a set of genusspecific primers for Fasciola ITS-2 amplification, and we designed species-specific internal TaqMan probes to identify F. hepatica and F. gigantica, as well as the hybrid ‘intermediate’ Fasciola. These primers and probes were used for the highly specific, sensitive, and simple identification of Fasciola species collected from different animal host from China, Spain, Niger and Egypt. The novel qPCR-based technique for the identification of Fasciola spp. may provide a useful tool for the epidemiological investigation of Fasciola infection, including their intermediate snail hosts.

This study aimed to finding possible relationship between epimastigote stage infecting Cephalopina titillator and Trypanosoma evansi infecting camels in Egypt. Nine rats were assigned into three equal groups and used for experimental infection. Rats of group 1 were inoculated intraperitoneally with 0.5ml of camel blood infected with T. evansi. Group 2 was inoculated intraperitoneally with 0.5ml of the fluid content of 3rd stage larvae of C. titillator infected with epimastigote stage. The 3rd group was kept as non-infected control. Blood smears were obtained daily from each rat of the experimental and control rats. Rats in group 1 showed the parasites in their blood on the 2nd day post infection (DPI) until the end of experiment 1month post infection (P.I.). The rats of group 2 and 3 showed no trypomastigote in their blood for a period of 1 month post inoculation. Sera were obtained from 1st, 2nd and 3rd groups at weekly interval starting from 1st week for a period of 4 weeks. The sera were tested using test kit CATT/ T. evansi. The sera of rats in group1 showed positive results in CATT from the 1st to the 4th week post inoculation. The rats of group 2 were positive for CATT at 3rd week and 4th week PI. While rats in group3 were negative for CATT during the experiment. Genomic DNA from purified trypanosomes and the fluid of 3rd stage larvae of C. titillator was extracted and DNA subjected to PCR technique. Also, PCR products of the two samples were purified from gel and the insert was sequenced and analyzed. The DNA from epimastigote form of C. titillator and that of T. evansi were positive for T. evansi by PCR using specific primer at 227bp. The result of gene sequence showed that the region of gene 227bp was identical in T. evansi and epimastigote stage obtained from C. titillator.

The present study provides clinical and laboratory examination of 54 cattle for infection by Theileria and Babesia piroplasmids. Prevalence of the clinical signs revealed high percentage of the infected animals showed pale mucous membrane (42.59%) and fever (38.88%), while low percentage showed swelling of lymph nodes and hematuria (18.51%). PCR result discovered the majority of Babesia spp infection (26%), most of them infected by B. bigemina (17%). The percentages of positive animals for Theileria annulata were 13% of total 22% Theileria infection. The use of PCR resulted in significantly higher efficacy of detection of bovine piroplasmids compared to microscopical examination of blood smears and allowed the specific discrimination between pathogenic and non-pathogenic Theileria spp which cannot be accomplished by traditional diagnosis. ELISA revealed higher babesiosis and theileriosis infection percentage (39% and 28% respectively) than that of PCR. The hematological and biochemical changes are demonstrated in this study.