Characterization of Sarcocystis Species Based on Traditional and Molecular Methods in Imported Frozen Buffalo Meat in Egypt

Abstract

Sarcocystis is an intracellular, cyst forming and worldwide distributed parasites. In the present study, fifty five different cuts of imported frozen buffalo meat were collected from different localities of Alexandria markets and were subjected to macroscopic and microscopic examination followed by molecular methods. Polymerase chain reaction used to amplify 18S rRNA gene from the collected samples followed by sequence analysis to detect infections with different Sarcocystis spp. The result revealed that the infection rate in examined sample was 70.9% by molecular methods; while it was 23% and 20% by macroscopic and microscopic examination respectively. Also the percentage of infection was varied among different examined cuts. Comparison of the obtained sequences with published data at the Genebank showed that frozen buffalo meat has multiple sarcocystis infections with S. fusiformis and S.cruzi. This study supported previous findings indicated that water buffalo could be infected with S.cruzi which produced microscopic cyst that has hygienic importance for both veterinary and medical aspects. SO, we recommended to used molecular method for detection sarcocystis infection especially in imported frozen meat to avoid human infection of such zoonotic parasite.

PREVALENCE AND GENETIC CHARACTERIZATION OF Sarcocystis fusiformis IN WATER BUFFALOES (Bubalus bubalis) IN TWO NORTHERN PROVINCES OF EGYPT

“…Cysts were separated from the surrounding muscles and genomic DNA was extracted independently from each of Sarcocystis cysts from the esophagus (five cysts from each province) using the Genomic DNA Extraction Kit (Thermo Fisher Scientific Inc., MA, USA) (24). The concentration of DNA was measured using a spectrophotometer (Thermo Fisher Scientific, USA).…”

“…The concentration of DNA was measured using a spectrophotometer (Thermo Fisher Scientific, USA). PCR reactions were applied using the primer set: forward (5´-GGCCCTTTTAGTGAGGGTGT-3´) and reverse (5´-TACGAATGCCCCCAACTGTC-3´) of the 18SrR-NA gene of S. fusiformis targeting 270bp fragment (24). PCR reactions were performed in a final reaction volume of 25 μL using 2× PCR Master Mix Solution (i-Taq™) (iNtRON Biotechnology, Seoul, Korea).…”

“…Negative control Double-distilled water was performed with all the reactions. The PCR reaction conditions were similar to previous research (24). Five μL of PCR products were tested by running on a 2.5% agarose gel (Sigma-Aldrich, USA), stained with ethidium bromide (Sigma-Aldrich, USA), using a 100-bp genetic marker (Sigma-Aldrich, USA), then visualized using a UV and photographed.…”

Molecular identification using 18S ribosomal RNA of Sarcocystis spp. in bovine minced meat in Van Province, Turkey

“…In particular, the 18S rRNA gene region is more widely used for the identification of Sarcocystis spp. than the other gene regions (4,16,18,23,24,27,33,42). However; it has been reported that S. hominis, S. bovifelis, S. bovini, and S. rommeli species cannot be distinguished by the 18S rRNA gene region, and the term 'S.…”

Survey on Sarcocystis infection in imported male cattle carcasses slaughtered at Duhok abattoir, Kurdistan region of Iraq

“…The high rate of infection and presence of different developmental cyst merozoite generation phase is regarded due to the contact and graze of the imported cattle for at least 2-3 months in contaminated pasture before slaughtering as a most common prevalent habit among livestock animals' owners in Kurdistan region however the pasture may contain sever amount of shaded oo-sporo/cyst (Fayer 1977;Khoshsima et al 2018). Mousa et al (2016) diagnosed 70% of the frozen meat came from Brazil and India infected with Sarcocystis by molecular techniques.…”