Browsing by Author "Halidi, Ahmet Galip"

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Investigation of Mitochondrial Cytb Gene Region of Both Echinococcus Granulosus Eggs From Dogs and Cystic Echinococcosis Isolates Obtained From Sheep and Cattle by Molecular Methods
(Iranian Scientific Society Medical Entomology, 2024) Yildiz, Rahmi; Aydemir, Selahattin; Halidi, Ahmet Galip; Unlu, Ahmet Hakan; Yilmaz, Hasan
Background:We aimed to determine the common Echinococcus granulosus genotypes in A & gbreve;r & imath;, T & uuml;rkiye and to obtain information on the transmission of this parasite. Methods:Cystic echinococcosissamples from 100 slaughtered cattle and 100 slaughtered sheep and faecal samples from 200 stray dogs were included in 2021. Collected cyst fluid samples and faces were examined microscopically. DNA was isolated from the germinal membrane of the cysts and from the parasite eggs in the stool samples. The mitochondrial cytbgene region of the parasite was amplified by PCR. Genotypes were determined using the Basic Local Alignment Search Tool (BLAST) after sequence analysis of PCR amplicons. Results:The highest percentage of cysts was found in the lungs of sheep and the liver of cattle. In addition, 75% of sheep cysts and 25.6% of cattle cysts were fertile. Taenia spp./Echinococcus spp. eggs werefound in 6% of the faeces of 200 dogs ex-amined microscopically. E. granulosuseggs were detected in 4 out of 50 stool sam-ples analysed by PCR. All samples analysed by sequence analysis were identified as E. granulosuss.s. G1 genotype. Sequence comparisonrevealed revealed one or more-pointmutations in different regions of the five samples. Conclusion:E. granulosuss.s. G1 genotype, known as sheep strain, is common in the A & gbreve;r & imath;, T & uuml;rkiye. The controlled slaughter of livestock, especially sheep, and the avoidance of feeding hydatid cyst organs to dogs, together with public education, were necessary to prevent the spread of the disease.
Investigation of the Effect of Pasteurization on the Viability of Cryptosporidium Parvum in Cow's Milk by Propidium Monoazide Qpcr
(Ankara Microbiology Soc, 2023) Aydemir, Selahattin; Durmaz, Hisamettin; Aydemir, Mehmet Emin; Kilic Altun, Serap; Demir, Abdulbaki; Halidi, Ahmet Galip; Arslan, Ali
Cow's milk, which is one of today's most important food sources, can be a reservoir for many patho-gens that create a risk to public health. One of these pathogens is Cryptosporidium parvum. The oocysts of C.parvum, an obligate intracellular parasite, cause infection when ingested orally. The oocysts scattered around with the feces of infected cows or calves can contaminate raw milk and this is frequently seen in dairy farms. The aim of this study was to investigate the viability of C.parvum by propidium monoazide (PMA)-quantitative polymerase chain reaction (qPCR) method after heat treatment applied to contami- nated raw cow's milk. For the study, 50 ml of unpasteurized cow's milk was contaminated with 5 X 105 C.parvum oocysts and portioned into 1.5 ml microcentrifuge tubes. Three groups, namely the control group, pasteurization group and boiling group were formed. No warming procedure was applied to the control group. In the pasteurization group, the milks in microcentrifuge tubes were poured into the wells of the dry block heater set to 71.7 degrees C and incubated for five seconds. At the end of the period, the milks were transferred to the wells of the cold metal tube, which was removed at-20 degrees C with the help of a micropipette, and incubated for five seconds. The milks in the boiling group were incubated for two minutes in a dry block heater set to 95 degrees C. After the heat treatment, the milks in microcentrifuge tubes were transferred to 10 ml centrifuge tubes, PBS was added to make the final volume 10 ml, and centrifuged at 4000 rpm for 20 minutes. After this process was repeated twice, 400 mu l of PBS was added to the precipitate remaining at the bottom, and the precipitate was homogenized. One sample of each group was applied with PMA, while PMA was not applied to the other sample. PMA-applied samples were incubated for five minutes at room temperature and in the dark, and then exposed to UV light for five minutes in the device with cooling feature. The oocysts were collected by centrifugation at 5000 g for five minutes. After DNA isolation from oocysts, SYBR Green real time PCR (Rt-PCR) was performed using primers amplifying the COWP gene region. As a result of SYBR Green Rt-PCR, the mean Ct values of the control without PMA, pasteurization and boiling groups were determined as 25 +/- 1.24, 23 +/- 0.98 and 26 +/- 1.03, respectively. While no peak was obtained in the boiling group after PMA application, the mean Ct values of the control and pasteurization groups were 28 +/- 1.38 and 31 +/- 1.46, respectively. As a result, it was concluded that live C.parvum cysts in milk could be detected by PMA-qPCR method and live oocysts could be found in pasteurized milk.