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.

Background: We aimed to determine the common Echinococcus granulosus genotypes in Agri, Türkiye and to obtain information on the transmission of this parasite. Methods: Cystic echinococcosis samples 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 cytb gene 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 were found in 6% of the faeces of 200 dogs examined microscopically. E. granulosus eggs were detected in 4 out of 50 stool samples analysed by PCR. All samples analysed by sequence analysis were identified as E. granulosus s.s. G1 genotype. Sequence comparison revealed revealed one or more-point mutations in different regions of the five samples. Conclusion: E. granulosus s.s. G1 genotype, known as sheep strain, is common in the Agri, Türkiye. 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.

The Significance of Opportunistic Parasitosis and Blastocystosis in Patients with Gastric Cancer: a Study with Control Group.

Objective: The aim of this study was to determine the prevalence of opportunistic parasites and Blastocystis spp. in patients with gastric cancer (CA) and to determine the significance of these parasite. Methods: The patient group and the control group were composed of 100 people each. The stool samples were examined under the microscope for intestinal parasites with the native-Lugol method. Then, samples were multiplied by formol-ethyl acetate method and stained with modified acid-fast method. Results: Intestinal parasite positivity was indicated in 14% of the gastric CA, and 2% of the healthy individuals (p=0.001). Blastocystis spp. (p=0.009) was identified in 11%, Cryptosporidium spp. was identified in 4%, G. intestinalis was identified in 2%, and C. cayetanensis was identified in 1% of the patient group. There were significant differences between the intestinal parasite positivity (p=0.012), abundant Blastocystis spp. positivity (p=0.041) and all Blastocystis spp. positivity (p=0.037) in patient and control groups. Most of the patients who were positive for parasites had diarrhea. Conclusion: Based findings, it was concluded that it would be beneficial to evaluate gastric CA patients, especially those with diarrhea, for intestinal parasites.

[Investigation of the Effect of Pasteurization on the Viability of Cryptosporidium parvum in Cow's Milk by Propidium Monoazide qPCR]. / Pastörizasyonun Inek Sütündeki Cryptosporidium parvum Canliligi Üzerine Etkisinin Propidium Monoazide qPCR ile Arastirilmasi.

Cow's milk, which is one of today's most important food sources, can be a reservoir for many pathogens 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 contaminated 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 °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 °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 °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 µl 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.

Investigation of Giardia spp., Cryptosporidium spp. and Cyclospora cayetanensis in Samples Collected from Different Spring Waters Igdir, Türkiye

OBJECTIVE: In this study, it was aimed to investigate the physical and chemical properties of different spring waters and parasitic factors with different methods. METHODS: This study was carried out on 69 water samples collected from different spring waters in and around Igdir Province in April and June 2021. The samples were analyzed by native-Lugol, modified acid-fast staining, and nested polymerase chain reaction (nPCR). In addition, altitude (meter) and pressure (mmHg) measurements were made at the point where the water samples were taken. RESULTS: One or more parasites were detected in 27.5% of the 69 water samples examined. Only C. cayetanensis was found in 13% of the samples, only Cryptosporidium spp. in 10.1%, only Giardia spp. in 1.4%, only C. cayetanensis and Giardia spp. in 1.4%, only C. cayetanensis, Cryptosporidium spp., and Giardia spp. in 1.4%. Only Giardia spp. cyst (4.3%) was detected by the direct examination method. While C. cayetanensis and Cryptosporidium spp. oocysts were detected in 8.7% and 7.2% of the samples by the modified acid-fast staining method, C. cayetanensis was detected in 15.9% and Cryptosporidium spp. was detected in 11.6% of the samples by nPCR. When the C. cayetanensis and Cryptosporidium spp. positivity rates were compared according to the characteristics of the water, there was no statistical difference between the altitude, salinity, pH, mmHg, and temperature (kelvin) values, but a significant correlation was found between the amount of dissolved oxygen and Cryptosporidium spp. positivity (p=0.047). CONCLUSION: Cryptosporidium spp., C. cayetanensis, and G. intestinalis are important waterborne pathogens that can cause epidemics. It is our belief that in order to reduce the risk of contamination of these parasitic factors with spring waters, public awareness should be raised, infrastructures should be improved, and new water treatment techniques, such as ultraviolet, ozonation and monitoring systems, should be used.

The Importance of Antioxidant Enzymes and Oxidative Stress in Human Fascioliasis

Objective: The aim of this study was to determine the levels of superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), and malondialdehyde (MDA) in the patients infected with Fasciola hepatica and establish whether these parameters differ among the patients with fascioliasis. Methods: The patient group consisted of 140 individuals with F. hepatica seropositive; the control group consisted of 140 healthy individuals who tested negative for this parasite and had no other diseases. The patient group consisted of individuals with no chronic diseases other than fascioliasis; in both the patient and the control groups, the subjects had no unhealthy habits such as smoking and alcohol consumption, etc. The blood samples taken to diagnose fascioliasis were evaluated by the ELISA method. The samples were studied according to the kit procedures for SOD, CAT, GPx and MDA markers. Results: In this study, 43.6% of 140 individuals in the patient group infected with F. hepatica had CAT (p=0.001), 35% had GPx (p=0.001), 12.9% had SOD (p=0.002), 90.7% had MDA (p=0.001). There was found a statistically significant difference between the patient and the control group in terms of the positivity of these four parameters. Conclusion: As a result, a statistically significant relationship was found between the increase in the SOD, GPx, CAT, and MDA levels and fascioliasis. The high rate of MDA revealed that oxidative stress occurred in patients with fascioliasis, resulting in an increased activity of SOD, GPx, and CAT. Amaç: Bu çalismanin amaci, Fasciola hepatica ile enfekte hastalarda süperoksit dismutaz (SOD), glutatyon peroksidaz (GPx), katalaz (CAT) ve malondialdehit (MDA) düzeylerini belirlemek ve fascioliasisli hastalarda bu parametrelerde farklilik meydana gelip gelmedigini ortaya koymaktir. Yöntemler: Hasta grubu, F. hepatica pozitif olan 140 hastadan; kontrol grubu ise bu parazit yönünden negatif bulunan ve baska herhangi bir hastaligi bulunmayan 140 saglikli kisiden olusturuldu. Hasta grubuna fascioliasis disinda herhangi bir kronik hastaligi olmayan ve hem hasta hem de kontrol grubuna sigara, alkol kullanimi olmayan kisiler dahil edildi. Hastalarda fascioliasis pozitifligini belirlemek için alinan kan örnekleri ELISA yöntemi ile çalisildi. Serum SOD, CAT, GPx ve MDA düzeyleri ELISA yöntemi ile degerlendirildi. Bulgular: Bu çalismada F. hepatica ile enfekte hasta grubundaki 140 kisinin %43,6'sinda CAT (p=0,001), %35'inde GPx (p=0,001), %12,9'unda SOD (p=0,002) ve %90,7 MDA pozitifligi saptandi. Bu dört parametrenin pozitifligi açisindan hasta ve kontrol grubu arasinda istatistiksel olarak anlamli fark bulundu (p=0,001). Sonuç: Sonuç olarak SOD, GPx, CAT ve MDA düzeyindeki artis ile fascioliasis arasinda istatistik olarak anlamli bir iliski saptanmistir. Çalismamizda MDA'nin yüksek oranda saptanmis olmasi fascioliasisli hastalarda oksidatif stres olustugunu ve SOD, GPx ve CAT aktivitelerinde artis oldugunu ortaya koymustur.

Genotyping of Echinococcus granulosus Human Isolates Obtained from the Puncture, Aspiration, Injection, and Re-Aspiration (PAIR) Process in Türkiye.

Background: Echinococcus granulosus is spread by the excretion of cystic organs into the environment. The dog is infected via eating the cystic organ. It then contaminates the environment with eggs of E. granulosus, which are infective to humans and animals. We aimed to determine the E. granulosus genotypes that cause infection in humans in the Van region, Türkiye. Methods: Sixty patients between 18 and 100 years of age, who underwent the puncture, aspiration, injection, re-aspiration (PAIR) procedure in the Department of Radiodiognastics of Van Yüzüncü Yil University, Van, Türkiye were included in the study. PAIR fluids were examined microscopically and DNA was isolated from the fluids. After DNA isolation, polymerase chain reaction (PCR) was performed using primers that amplify the E. granulosus NADH dehydrogenase subunit 1 (NAD1) gene region. After sequence analysis of the PCR amplicons, Basic Local Alignment Search Tool (BLAST) was performed. Results: In the microscopic examination, protoscolex or hook was detected in 42 (70%) of the samples. DNA was successfully extracted from all of the cyst fluids containing protoscolex and hook, and the NAD1 gene region was PCR-amplified. After using BLAST, all of the samples were determined to be an E. granulosus sensu stricto G1 genotype. Sequence comparison revealed that four (9.5%) isolate sequences showed single nucleotide polymorphism (SNP). Sequences of isolates with SNP submitted to the GenBank with accession numbers OR565864 to OR565867. Conclusion: E. granulosus s.s. G1 genotype, known as sheep strain, is common in human hydatid disease in the Van region of Türkiye.

Investigation of Pneumocystis jirovecii in Lung Cancer Patients with the Nested PCR Method

Objective: Pneumocystis jirovecii (P. jirovecii) is an opportunistic pathogen in humans. Early diagnosis and optimal treatment of patients with P. jirovecii pneumonia (PJP) remains a key priority. This study investigated P. jirovecii in patients with lung cancer using the nested-polymerase chain reaction (PCR) method and examined the relationship between P. jirovecii and clinical findings. Methods: The study included 60 patients with lung cancer and 30 patients without lung cancer. The bronchoalveolar lavage (BAL) fluid samples of these 90 individuals were taken for diagnostic purposes in the University of Health Sciences Turkey, Van Training and Research Hospital, Clinic of Chest Diseases. Patient information was recorded. After DNA isolation from the BAL fluid samples taken from patients, the nested-PCR protocol for amplification of mtLSUrRNA in P. jirovecii was performed. Results: P. jirovecii DNA was detected in 40 (66.67%) of the lung cancer patients included in the study and in six (20%) patients without lung cancer, that is, in 46 (51.11%) patients. The rate of nested-PCR positivity in the lung cancer group was significantly higher than that in the non-lung cancer group (p=0.0001). Additionally, a statistically significant correlation was found between anorexia and weight loss, fever and sputum P. jirovecii positivity in patients with lung cancer (p<0.005). Conclusion: These findings suggest that lung cancer patients should be evaluated for PJP.

Effects of degree of translation or rotation of acetabular fragment of periacetabular osteotomy procedure on pelvic X-ray parameters.

The present study aims to investigate the effect of amount of lateralization and/or anteversion of the point where the iliac cut meets with the posterior column cut of periacetabular osteotomy (PAO), on X-ray parameters such as Center of edge (CE) angle, retroversion index (RVI) and sharp angle. Fourteen patients with symptomatic hip dysplasia (CE° < 20°) were included. Pelvis Computerized tomography (CT) sections were used for 3D printing. PAO was then performed on these models. The point (A), 1 cm lateral to the pelvic brim, is marked where the iliac cut intersects the posterior column cut. In Group I (1.5-0), point A is lateralized parallel to the osteotomy line for 1.5 cm. In Group II (1.5-0.5), it is additionally anteverted for 0.5 cm. In Group III (3-0), point A is lateralized for 3 cm and then additionally anteverted for 1 cm (Group IV: 3-1). Radiographs were taken in each stage. The lateral CE angle, RVI and sharp angle were measured. All had an increase in the CE angle and RVI and a decrease in the sharp angle compared to the control group (P < 0.05). The amount of CE angle (ΔCE) or RVI increase (ΔRV) was as follows: 3-1(38°, 0.3) > 3-0(27°, 0.2) and 1.5-0.5(25°, 0.1) > 1.5-0(17°, 0.07) (P < 0.05) (with no difference between groups 1.5-0.5 and 3-0, P = 0.7). The amount of sharp angle decrease was as follows: 3-1(20°), 3-0(18°) < 1.5-0.5(11°) < 1.5-0(8°) (P < 0.05). The lateralization of the intersection point where the iliac wing cut meets with the posterior column cut along the cut surface led to an increase of lateral cover and focal retroversion. Additional anteversion leads to further increases in those parameters, while groups 1.5-0.5 and 3-0 did not differ between.

Local Steroid Treatment: An Effective Procedure for Idiopathic Granulomatous Mastitis, Including Complicated Cases.

PURPOSE: To evaluate the effectiveness of treatment with topical and intralesional steroids for idiopathic granulomatous mastitis (IGM) and to compare with surgical methods. METHODS: Data were retrospectively collected from records. Intralesional steroid injection and topical steroid administration, hereafter referred to as local steroid treatment (LST) were applied in Group 1. Surgery (local excision, wide excision, and mastectomy) was performed in Group 2. In Group 1, changes in lesion sizes were recorded and factors complicating treatment were identified. The Numeric Pain Rating Scale was used to determine subjective pain. LST and surgery were compared with regard to: pain before and after the treatment; complication rate; recurrence rate; and treatment cost. RESULTS: There were 38 and 48 patients in Group 1 and Group 2, respectively. In the LST group, 72 lesions were present and 70 of 72 (97%) responded completely to treatment. Pretreatment median maximum diameter was 23.50 (15.25-35.25) mm, which regressed to 16 (12-25) mm after the first session. While the pretreatment pain scores of Group 1 and Group 2 were similar (p = 0.756), there was a significant difference in the post-treatment pain scores (p < 0.001). No recurrence occurred in any patients in Group 1, while recurrence developed in 15 (31.2%) patients in Group 2 (p < 0.001). CONCLUSION: LST is a treatment for IGM that is cheap, with high efficiency, negligible recurrence, and has good esthetic outcome. Our results suggest that LST should be the first-line treatment option for all IGM patients, including complicated cases.

Genetic Diversity of Echinococcus granulosus Isolated from Humans: A Comparative Study in Two Cystic Echinococcosis Endemic Areas, Turkey and Iran.

Cystic echinococcosis (CE) is one of the most important zoonotic parasitic diseases caused by the larval stage of Echinococcus granulosus. Based on molecular studies and DNA sequencing, E. granulosus has been classified into 10 different genotypes (G1 to G10). Two neighboring countries, Turkey and Iran, are considered the two main foci of CE in the Middle East. The current study is aimed at examining the genotype diversity of E. granulosus isolated from human clinical samples in Turkey and Iran. Surgically removed human hydatid cysts were collected from East Azerbaijan and Fars provinces in Iran and Van province in Turkey. After extracting DNA, performing PCR, targeting the cox1 gene, the PCR products were purified from the gel and were sequenced from both directions. The sequences were aligned and compared, using BioEdit and also the BLAST program of GenBank. The maximum likelihood tree was constructed based on the Tamura-Nei model, using the MEGAX software. Phylogenetic analysis showed that the human isolated samples were classified into two major clades: G1 (from Iran and Turkey) and G3 (5 samples from northwestern Iran and one sample from Turkey). The mean and degree of genetic divergence (K2P) between the two major clades, G1 and G3, were 0.2% and 0.7 ± 0.4%, respectively. The findings of the current study revealed that the sheep strain (G1) and the less important strain G3 have major roles in the transmission cycle of CE in two neighboring countries, Iran and Turkey. Therefore, it is necessary to interpose the life cycle of this parasite and reduce the disease burden in livestock and humans by adopting common regional preventive and control policies.