Hilali, M., and A. Mohamed, "The dog (Canis familiaris) as the final host of Sarcocystis cameli (Mason, 1910).", Tropenmedizin und Parasitologie, vol. 31, issue 2, pp. 213-4, 1980 Jun.
Fereiga, R. M., M. R. AbouLaila, S. G. A. Mohamed, H. Y. A. H. Mahmoud, A. O. Ali, A. F. Ali, M. Hilali, A. Zaid, A. E. A. Mohamed, and Y. Nishikawa, "Serological detection and epidemiology of Neospora caninum and Cryptosporidium parvum antibodies in cattle in southern Egypt", Acta Tropica, 2016. Abstract

Neospora caninum and Cryptosporidium parvum are intracellular protozoan parasites that are distributed worldwide and of major economical concern in cattle industry. N. caninum can cause abortion storms and high culling rates, whereas C. parvum has zoonotic implications and can cause diarrhea in calves. There are currently no data on the prevalence of neosporosis and cryptosporidiosis in humans or animals in southern Egypt. Prevalence of these two infections was determined in a sample of cattle from two different areas in southern Egypt, Sohag and Qena, using enzyme-linked immunosorbent assay. A total 301 cattle were sampled, of which 18.9% were positive for N. caninum, 35.9% were positive for C. parvum and 10.0% were positive for both. Geographical location and breeding system were considered as potential risk factors for C. parvum infection. A higher prevalence of infection was identified on small scale farms, compared with larger, intensive systems, with a prevalence of 50.2% compared with 37.8%, respectively. Animals in Sohag had a significantly higher prevalence compared with Qena, with a seroprevalence of 46.1% compared with 31.6%, respectively. In brief, marked seroprevalence recorded in this study indicates a high incidence of N. caninum and C. parvum infections in cattle, and this necessitates the application of more effective strategies for combating these types of infections on farms in Egypt.

CpP23, immunodominant surface glycoprotein of Cryptosporidium parvum; GST, glutathione-S transferase; NcSAG1t, Neospora caninum truncated surface antigen 1

ElBehairy, A. M., S. Choudhary, L. R. Ferreira, O. C. H. Kwok, M. Hilali, C. Su, and J. P. Dubey, "Genetic characterization of viable Toxoplasma gondii isolates from stray dogs from Giza, Egypt.", Veterinary parasitology, vol. 193, issue 1-3, pp. 25-9, 2013 Mar 31. Abstract

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.

Rasche, A., M. Saqib, A. M. Liljander, S. Bornstein, A. Zohaib, S. Renneker, K. Steinhagen, R. Wernery, M. Younan, I. Gluecks, et al., "Hepatitis E Virus Infection in Dromedaries, North and East Africa, United Arab Emirates, and Pakistan, 1983-2015.", Emerging infectious diseases, vol. 22, issue 7, pp. 1249-52, 2016 Jul. Abstract

A new hepatitis E virus (HEV-7) was recently found in dromedaries and 1 human from the United Arab Emirates. We screened 2,438 dromedary samples from Pakistan, the United Arab Emirates, and 4 African countries. HEV-7 is long established, diversified and geographically widespread. Dromedaries may constitute a neglected source of zoonotic HEV infections.

Gjerde, B., and M. Hilali, "Domestic cats (Felis catus) are definitive hosts for Sarcocystis sinensis from water buffaloes (Bubalus bubalis).", The Journal of veterinary medical science / the Japanese Society of Veterinary Science, 2016 Apr 14. Abstract

The definitive hosts of Sarcocystis sinensis in water buffaloes have hitherto been unknown, but the close similarity of this species to the cat-transmitted Sarcocystis bovifelis in cattle suggested they were felids. In a previous study, two domestic cats were fed macroscopic sarcocysts of Sarcocystis fusiformis contained within or dissected from the esophageal muscles of water buffaloes, while no microscopic sarcocysts of S. sinensis were noticed. Both cats started shedding small numbers of sporocysts 8-10 days post infection (dpi) and were euthanized 15 dpi. Using a PCR-based molecular assay targeting the mitochondrial cox1 gene of S. fusiformis, both cats were shown to act as definitive hosts for this species. In the present study, DNA samples derived from oocysts/sporocysts in the intestinal mucosa of both cats were further examined by PCR for the presence of S. sinensis using 2 newly designed primers selectively targeting the cox1 gene of this species. All 6 DNA samples examined from each cat tested positive for S. sinensis. A 1,038-bp-long portion of cox1 was amplified and sequenced as 2 overlapping fragments from 5 of these DNA samples. The 5 sequences shared 99.3-100% identity with 7 previous cox1 sequences of S. sinensis obtained from sarcocysts in water buffaloes. Additionally, amplification of the ITS1 region with primers targeting various Sarcocystis spp., yielded amplicons of 2 different lengths, corresponding to those obtained from sarcocyst isolates of S. sinensis and S. fusiformis, respectively. This is the first study to show that cats act as definitive hosts for S. sinensis.

Dubey, J. P., E. Van Wilpe, S. K. Verma, and M. Hilali, "Ultrastructure of Sarcocystis bertrami sarcocysts from a naturally infected donkey (Equus asinus) from Egypt.", Parasitology, vol. 143, issue 1, pp. 18-23, 2016 Jan. Abstract

There is considerable confusion concerning Sarcocystis species in equids. Little is known of Sarcocystis infections in donkeys (Equus asinus). Here we describe the structure of Sarcocystis bertrami-like from the donkey by light microscopy (LM) and transmission electron microscopy (TEM). Nineteen sarcocysts from the tongue of a donkey from Egypt were studied both by LM and TEM. By LM, all sarcocysts had variably shaped and sized projections on the sarcocyst walls, giving it a thin-walled to thick-walled appearance, depending on individual sarcocyst and plane of section. By TEM, sarcocysts walls had villar protrusions (vp) of type 11. The sarcocyst wall had conical to slender vp, up to 6 µm long and 1 µm wide; the vp were folded over the sarcocyst wall. The total thickness of the sarcocyst wall with ground substance layer (gs) was 1-3 µm. The vp had microtubules (mt) that originated deeper in the gs and continued up to the tip. The apical part of the vp had electron dense granules. The mt were configured into 3 types: a tuft of electron dense mt1 extending the entire length of the vp with a tuft of medium electron dense mt2 appearing in parallel, and fine mt3 present only in the villar tips. The gs was mainly smooth with few indistinct granules. All sarcocysts were mature and contained metrocytes and bradyzoites. Bradyzoites were approximately 11-15 × 2-3 µm in size with typical organelles.

Gjerde, B., M. Hilali, and S. A. Mawgood, "Molecular characterisation of three regions of the nuclear ribosomal DNA unit and the mitochondrial cox1 gene of Sarcocystis fusiformis from water buffaloes (Bubalus bubalis) in Egypt.", Parasitology research, vol. 114, issue 9, pp. 3401-13, 2015 Sep. Abstract

A total of 33 macroscopically visible (3-11 × 1-5 mm) sarcocysts of Sarcocystis fusiformis were excised from the oesophagus of 12 freshly slaughtered water buffalos in Giza, Egypt. Genomic DNA was extracted from the sarcocysts, and all isolates were characterised at the mitochondrial cytochrome c oxidase subunit I (cox1) gene through PCR amplification and direct sequencing, whereas a few selected isolates were characterised at the 18S and 28S ribosomal (r) RNA genes and the internal transcribed spacer 1 (ITS1) region of the nuclear rDNA unit following cloning. Among the 33 cox1 sequences (1,038-bp long), there was a total of 13 haplotypes, differing from each other by one to seven substitutions and sharing an identity of 99.3-99.9 %. In comparison, the sequence identity was 98.8-99.0 % among eight complete 18S rRNA gene sequences (1,873-1,879-bp long), 98.1-100 % among 28 complete ITS1 sequences (853-864-bp long) and 97.4-99.6 % among five partial 28S rRNA gene sequences (1,607-1,622 bp). At the three nuclear loci, the intraspecific (and intra-isolate) sequence variation was due to both substitutions and indels, which necessitated cloning of the PCR products before sequencing. Some additional clones of the 18S and 28S rRNA genes were highly divergent from the more typical clones, but the true nature of these aberrant clones could not be determined. Sequence comparisons and phylogenetic analyses based on either 18S rRNA gene or cox1 nucleotide sequences, placed S. fusiformis closest to Sarcocystis cafferi from the African buffalo, but only the analyses based on cox1 data separated the two taxa clearly from each other and showed that they were separate species (monophyletic clusters and 93 % sequence identity at cox1 versus interleaved sequences and 98.7-99.1 % sequence identity at the 18S rRNA gene). Two cats experimentally infected with sarcocysts of S. fusiformis started shedding small numbers of sporocysts 8-10 days post-infection (dpi) and were euthanized 15 dpi. Sporocysts isolated from the intestinal mucosa of both cats were identified molecularly as belonging to S. fusiformis through PCR amplification and sequencing of the partial cox1. The two sporocyst-derived cox1 sequences were identical with the most common sarcocyst-derived cox1 haplotype.

Dubey, J. P., M. Hilali, E. Van Wilpe, R. Calero-Bernal, S. K. Verma, and I. E. Abbas, "A review of sarcocystosis in camels and redescription of Sarcocystis cameli and Sarcocystis ippeni sarcocysts from the one-humped camel (Camelus dromedarius).", Parasitology, vol. 142, issue 12, pp. 1481-92, 2015 Oct. Abstract

There is considerable confusion concerning Sarcocystis species in camels. Five species: Sarcocystis cameli, Sarcocystis ippeni, Sarcocystis camelicanis, Sarcocystis camelocanis and Sarcocystis miescheri were named with inadequate descriptions and no type specimens. Here, we review literature on sarcocystosis in camels worldwide and redescribe structure of S. cameli and S. ippeni sarcocysts by light- and transmission electron microscopy (LM and TEM). Eight sarcocysts from the oesophagi of two camels (Camelus dromedarius) from Egypt were studied. By LM, all sarcocysts were thin-walled with barely visible projections on the cyst walls. By TEM, two structurally distinct sarcocysts were recognized by unique villar protrusions (vp) not found in sarcocysts from any other host. Sarcocysts of S. cameli had vp of type 9 j. The sarcocyst wall had upright slender vp, up to 3.0 µM long and 0.5 µM wide; the total thickness of the sarcocyst wall with ground substance (gs) layer was 3.5 µM. On each vp, there were rows of knob-like protrusions that appeared to be interconnected. The vp had microtubules that originated at midpoint of the gs and continued up to the tip; microtubules were smooth, without any granules or dense areas. Bradyzoites were approximately 14-15 × 3-4 µM in size with typical organelles. Sarcocystis ippeni sarcocysts had type 32 sarcocyst wall characterized by conical vp with an electron dense knob. The total thickness of the sarcocyst wall (from the base of gs to vp tip) was 2.3-3.0 µM. The vp were up to 1.2 µM wide at the base and 0.25 µM at the tip. Microtubules in vp originated at midpoint of gs and continued up to tip; microtubules were criss-crossed, smooth and without granules or dense areas. Bradyzoites were 12.0-13.5 × 2.0-3.0 µM in size. Sarcocystis camelicanis, S. camelocanis and S. miescheri are considered invalid.

Dubey, J. P., M. Hilali, E. Van Wilpe, S. K. Verma, R. Calero-Bernal, and A. Abdel-Wahab, "Redescription of Sarcocystis fusiformis sarcocysts from the water buffalo (Bubalus bubalis).", Parasitology, vol. 142, issue 2, pp. 385-94, 2015 Feb. Abstract

Four valid species of Sarcocystis have been reported from the water buffalo (Bubalus bubalis): Sarcocystis fusiformis, Sarcocystis buffalonis, Sarcocystis levinei and Sarcocystis dubeyi. Here, we redescribe structure of S. fusiformis sarcocysts by scanning and transmission electron microscopy (SEM, TEM). Twenty-one macroscopic sarcocysts from oesophagus of the water buffalo in Egypt were examined by light microscopy, SEM and TEM. The sarcocyst wall was up to 9 μm thick, depending on the section and the technique. In 5 μm paraffin-embedded sections, the sarcocyst wall was indistinct, 2-5 μm thick and appeared smooth. In 1 μm plastic-embedded sections stained with toluidine blue, the sarcocyst wall was 2.5-5.2 μm thick and had branched villar protrusions (vp)-like branches of a dead tree. By SEM, the sarcocyst wall had a mesh-like structure with irregularly shaped vp that were folded over the sarcocyst wall. On each vp there were uniform papillomatous structures that were 100 nm wide. By TEM, vp were up to 6 μm long and contained filamentous tubular structures, most of which were parallel to the long axis of the projections; granules were absent from these tubules. By TEM, bradyzoites within the same cyst varied from 11.2 to 16.8 μm in length. By TEM, bradyzoites had a very long (10 μm) convoluted mitochondrion, up to 12 dense granules, but only 2 rhoptries. This redescription should help to differentiate the sarcocysts of S. fusiformis from similar sarcocysts in domestic and wild ruminants.

Al-Kappany, Y. M., S. A. Abu-Elwafa, M. Hilali, B. M. Rosenthal, D. B. Dunams, and J. P. Dubey, "Experimental transmission of Sarcocystis muris (Apicomplexa: Sarcocystidae) sporocysts from a naturally infected cat (Felis catus) to immunocompetent and immunocompromised mice.", The Journal of parasitology, vol. 99, issue 6, pp. 997-1001, 2013 Dec. Abstract

Cats serve as definitive hosts for zoonotic Toxoplasma gondii , a protozoan that threatens human reproductive health, but they also excrete sporocysts of related protozoan that pose no known human health risk. Here we provide the first definitive evidence for natural infection with the enzootic parasite Sarcocystis muris, one such enzootic parasite. Sporulated Sarcocystis sp. sporocysts were found in rectal contents of an adult feral cat ( Felis catus ) in Giza, Egypt. After these sporocysts were orally inoculated into 2 Swiss Webster mice, sarcocysts were found to have developed in skeletal muscles 114 days later. As observed through transmission electron microscopy, the cyst wall corresponded to Type 1, and the parasitophorous vacuolar membrane had tiny outpocketing of blebs (<200 nm thick) that were not invaginated into the interior of the cyst; these structures were identical to the sarcocyst wall described for a Costa Rican isolate of S. muris that has served as an experimental model for nearly 4 decades. Two parasite-free cats fed sarcocyst-infected muscles developed patent infections; fully sporulated sporocysts (10-11 × 7.0 μm) were found in the lamina propria of small intestines of cats killed 6 and 7 days postinoculation (PI). Interferon gamma gene knockout (KO) mice were orally inoculated with sporocysts from experimentally infected cats, and their tissues were examined histologically; sarcocysts were found in 5 KO mice killed 87, 115, 196, 196, 196 days PI, but no stages were seen in 5 KO mice 10, 14, 14, 18, and 39 days PI. Bradyzoites were released from intramuscular sarcocysts of a KO mouse killed 115 days PI and orally inoculated into 5 KO mice. No stage of Sarcocystis was found in any organ (including intestinal lamina propria) of KO mice killed 4, 8, 81, 190, and 190 days PI, confirming that the definitive host is required to complete the life cycle even in the case of immunodeficient mice. This is the first confirmation of S. muris infection in a naturally infected cat anywhere.