Temperature and Time Dependent Variations in Anti-Toxoplasma gondii Antibody Titrations in Serum Samples

Objective: This study aimed to determine the temperature- and time-dependent variations in the anti-Toxoplasma gondii (T. gondii) antibody titres in serum samples collected from T. gondii-infected patients. Methods: During May 2017-February 2018, five serum samples (stored at different time periods and conditions) from pregnant or non-pregnant women aged ≥18 years who were infected with T. gondii and had applied to our Parasitology Department of Hafsa Sultan Hospital, Manisa Celal Bayar University, were investigated for the anti-T. gondii IgG antibody levels by enzyme linked fluorescent assay (ELFA). Results: The serum samples of five female volunteers who were infected with T. gondii that were stored at room temperature (20/25 °C), in a cargo package (+4/+8 °C), in a refrigerator (+4 °C), in a deep freezer (-16/-20 °C) and in an incubator (+37 °C) were tested at 0, 24, 48 and 72 hours after infection with the ELFA test. No statistically significant difference was observed in the anti-T. gondii IgG antibody titres (p>0.05). Conclusion: The results obtained from the patients infected with T. gondii at different times and conditions of up to 72 hours were not significantly affected clinically. Hence, more comprehensive data can be obtained by increasing the number of patients and storing the serum samples for more than 72 hours.

[Cystic Echinococcosis in Humans in Turkey]. / Türkiye'de Insanlarda Kistik Ekinokokkoz.

Although cystic echinococcosis (CE) is quite prevalent in Turkey, it is extremely neglected due to being usually asymptomatic for years and frequently not to be reported although it is obligatory. Most of the data on the prevalence of CE in humans in Turkey are based on hospital records, reported cases and the studies based on serological methods and they do not reflect the truth. The fact that detecting no cysts in most of the seropositive cases limits the value of researches based on serological tests. The most valuable epidemiologic data on CE are obtained by mass screening surveys with the use of portable ultrasonography (US) and it took the place of serological tests, especially in the last 20 years. Two of 190 cases older than 20 years were found to be positive for CE in a village of Konya city at the first study that US was performed in Turkey. At the first research performed on preliminary school children in Manisa Province; of the 630 students examined by US, serology and chest X-ray, 2 (0.3%) were diagnosed as CE by US. Only US was used at the second study, and hydatid cysts were observed in 3 (0.5%) of the 575 students in two villages; these data suggested that the use of US alone was more easy, fast and beneficial in the field studies. In the third research, 6093 students from 37 different schools of Manisa Province were selected as a representative sample, and 9 (0.2%) children (two previously operated) were found to be positive for CE by US. The only response to the invitation to use this model in different regions of Turkey was from Elazig Province and of the 2500 students selected, six cases (one previously operated) were detected, and the prevalence was found to be 0.2% in Elazig Province. During the same years, of the 102 cases sharing the same living space with 40 patients operated due to CE, 13 (12.7%) were radiologically diagnosed as CE in Van, while CE was diagnosed in 1 (0.5%) of the 209 cases in an area dealing with animal husbandry in Aydin. At the fourth research in Manisa, 4275 students from university were examined by US, while 2034 of these were also serologically tested by ELISA and Western Blot (WB). The efficacy of WB as a screening test in CE was investigated for the first time in the world; six new and three operated cases were detected, and the prevalence was 0.2%. During the research in the rural areas of Bulgaria, Romania and Turkey, of the 8618 cases living in six cities (Ankara, Aksaray, Balikesir, Bitlis, Edirne, Sanliurfa) of Turkey, 53 (0.6%) abdominal CE cases were detected by US and one of every 163 cases in Turkey was found to be infected with CE. This ratio shows that CE is one of the most important public health problems in Turkey. Control of CE is possible with "One Health" concept. An effective control program and changes in valid laws are needed in Turkey. In this review, the value of different diagnostic procedures have also been discussed.

Design, synthesis, and in vitro biological evaluation of novel thiazolopyrimidine derivatives as antileishmanial compounds.

A series of thiazolopyrimidine derivatives was designed and synthesized as a Leishmania major pteridine reductase 1 (LmPTR1) enzyme inhibitor. Their LmPTR1 inhibitor activities were evaluated using the enzyme produced by Escherichia coli in a recombinant way. The antileishmanial activity of the selected compounds was tested in vitro against Leishmania sp. Additionally, the compounds were evaluated for cytotoxic activity against the murine macrophage cell line RAW 264.7. According to the results, four compounds displayed not only a potent in vitro antileishmanial activity against promastigote forms but also low cytotoxicity. Among them, compound L16 exhibited an antileishmanial activity for both the promastigote and amastigote forms of L. tropica, with IC50 values of 7.5 and 2.69 µM, respectively. In addition, molecular docking studies and molecular dynamics simulations were also carried out in this study. In light of these findings, the compounds provide a new potential scaffold for antileishmanial drug discovery.

[In vitro susceptibility of Trichomonas vaginalis to metronidazole, ornidazole and proton pump inhibitors pantoprazole and esomeprazole]. / Trichomonas vaginalis'in metronidazol, ornidazol ve proton pompa inhibitörleri pantoprazol ve esomeprazole karsi in vitro duyarliligi.

The current treatment of trichomoniasis is based on the use of 5-nitroimidazole derivatives. Although metronidazole is reliable, inexpensive and highly effective against anaerobic microorganisms and protozoa, the development of metronidazole-resistant T.vaginalis strains pose to an increasing problem. Nitroimidazoles are compounds having azomycin (2-nitroimidazole) chemical structure and are obtained from Streptomyces strains. Benzimidazole, which is found in the structure of proton pump inhibitors, is also present in the other components that have antiprotozoal activity. In this study, the in vitro susceptibility of T.vaginalis against metronidazole, ornidazole, and the proton pump inhibitors which are tested recently as antiprotozoal agents; pantoprazole and esomeprazole was investigated. For this purpose a clinical T.vaginalis strain which was formerly isolated and stored after cryopreservation process in our laboratory was used. Minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) values of those agents against to this strain were determined in vitro by dilution method in 24-well cell culture plates. Trypticase yeast extract maltose medium, horse serum and antibiotic (penicillin + streptomycin) were distributed to each well of cell culture plates and after metronidazole, ornidazole, pantoprazole and esomeprazole solutions were added to two wells for each as 800, 400, 200, 100, 50 and 25 µg/ml, followed by the addition of 1 ml 5x10(3) T.vaginalis trophozoites into each well. Plates were incubated at 37°C, and viability and motility of the trophozoites were evaluated under light microscope at 24, 48 and 72 hours after incubation. MIC and MLC values of metronidazole/ornidazole in the 72(th) hour were found as 50 µg/ml and 100 µg/ml, respectively. MIC and MLC values for pantoprazole in the 72th hour were 200 µg/ml and 400 µg/ml, while the values for esomeprazole were 400 µg/ml ve 800 µg/ml, respectively. As a result, T.vaginalis strain used in the study was susceptible to metronidazole and ornidazole, besides, it was considered that pantoprazole and esomeprazole were also effective to the parasite and could be used as alternative drugs. However, further in vitro and clinical studies are clearly needed on the antiprotozoal effects of proton pump inhibitors. To our knowledge, this study was the first in literature, which esomeprazole's susceptibility on T.vaginalis was investigated in vitro.

Putative bronchopulmonary flagellated protozoa in immunosuppressed patients.

Flagellated protozoa that cause bronchopulmonary symptoms in humans are commonly neglected. These protozoal forms which were presumed to be "flagellated protozoa" have been previously identified in immunosuppressed patients in a number of studies, but have not been certainly classified so far. Since no human cases of bronchopulmonary flagellated protozoa were reported from Turkey, we aimed to investigate these putative protozoa in immunosuppressed patients who are particularly at risk of infectious diseases. Bronchoalveolar lavage fluid samples of 110 immunosuppressed adult patients who were admitted to the Department of Chest Diseases, Hafsa Sultan Hospital of Celal Bayar University, Manisa, Turkey, were examined in terms of parasites by light microscopy. Flagellated protozoal forms were detected in nine (8.2%) of 110 cases. Metronidazole (500 mg b.i.d. for 30 days) was given to all positive cases and a second bronchoscopy was performed at the end of the treatment, which revealed no parasites. In conclusion, immunosuppressed patients with bronchopulmonary symptoms should attentively be examined with regard to flagellated protozoa which can easily be misidentified as epithelial cells.

A mass screening survey of cystic echinococcosis by ultrasonography, Western blotting, and ELISA among university students in Manisa, Turkey.

Cystic echinococcosis (CE) is one of the most important zoonotic diseases in a wide geographic area, including Turkey. In the present project, a total of 4275 students from Celal Bayar University, Manisa, Turkey, were screened by ultrasonography (US) and specific antibodies for CE were examined by Western blotting (WB) and ELISA in finger prick blood samples of 2034 of 4275 volunteered students. We aimed to report the apparent prevalence of CE based on different diagnostic procedures and to compare WB and ELISA with US in diagnosis of CE in a mass screening setting. Six new cases were diagnosed as CE by US during the survey. In addition to these cases, three students were also detected to have been previously operated and pathologically confirmed for hepatic CE. US revealed parenchymal changes in these cases in concordance with their operation history; so, the prevalence of CE by US was calculated as 0.21% (9/4275) (95%CI, 0.11-0.39%) among university students in Manisa. Bands were detected at 8, 28, 32, 38, 42, 47, 70 and 90kDa by WB and the cases were considered to be positive for CE when at least three of the bands were seen together. Apparent prevalence of CE by ELISA and WB were found to be 2.11% (43/2034) (95%CI, 1.57-2.83%) and 0.25% (5/2034) (95%CI, 0.10-0.57%), respectively. Of the six US positive cases, WB was positive in only one case with two cysts in the liver. All of four cases with liver involvement were positive by ELISA. The high prevalence of CE among university students in Manisa indicated that CE is a major health problem in this area of Turkey. Our results supported that WB is rather difficult and not feasible as a mass screening test and may not be effective for confirmation especially in asymptomatic cases. As a result, we recommend US to be used initially in mass screening surveys for CE followed by confirmation by ELISA for suspected cases. Further examination primarily by chest X-ray followed by computed tomography and/or magnetic resonance imaging, if needed, should be recommended for US negative, ELISA and WB positive individuals who may have non-abdominal cysts.

[Evaluation of serological results of patients with suspected Toxoplasmosis admitted to the medical parasitology laboratory of Celal Bayar University Hospital between 2006-2010]. / Celal Bayar Üniversitesi Hastanesi Tibbi Parazitoloji Laboratuvarina 2006-2010 Yillarinda Toxoplasmosis Süphesi ile Basvuran Hastalarin Serolojik Sonuçlarinin Degerlendirilmesi.

OBJECTIVE: The aim of this study is to determine the serological results of patients with suspected toxoplasmosis who were admitted to the Medical Parasitology Laboratory of Celal Bayar University Hospital in Manisa between 2006 and 2010. METHODS: Anti-Toxoplasma gondii IgG and IgM antibodies were investigated by IFAT, IgG ELISA, and IgM Capture ELISA in 2815 patients. IgG avidity tests were also performed in cases with both IgG and IgM seropositivity. RESULTS: Only IgG, only IgM and both IgG and IgM seropositivity were detected in 657 (23.3%), 4 (0.1%) and 6 (0.2%) cases respectively among 2815 patients. Anti-T. gondii IgG seropositivity was mostly found in patients [21.2% (n=139)] who consume raw meat. IgG seroprevalence was found to be statistically higher in the 15-49 age group than the 0-14 age group (p < 0.05). Decrease in the prevalence of anti- T. gondii IgG seropositivity in 2010 was found to be statistically significant considering the five years period (p < 0.001). CONCLUSION: Raw meat consumption was detected as the most important risk factor in Toxoplasma infection. A decrease in anti-T. gondii IgG antibodies was detected in recent years considering the five years' serologic data, but toxoplasmosis remains important as a public health problem in Manisa.

[Incidence of intestinal parasites detected in the Department of Parasitology in Celal Bayar University Hospital between 2006 and 2010]. / Celal Bayar Üniversitesi Hastanesi Parazitoloji Poliklinigi'nde 2006-2010 Yillari Arasinda Saptanan Bagirsak Parazitlerinin Insidansi

OBJECTIVE: Results of 17.711 patien admitted to the Department of Parasitology in Celal Bayar University Hospital for parasitological stool examination between January 2006 and December 2010 were evaluated. METHODS: All stool samples were examined with wet mounts, formalin ethyl acetate concentration and trichrome staining methods. In addition, cellophane tape preparations from 5952 patients were evaluated. RESULTS: Intestinal parasites were detected in 2337 (13.12%) of 17.711 patients who were admitted during a five year period. The highest parasite infected individual ratio (16.69%) was found in 2008. The most frequently identified intestinal parasites were Blastocystis spp. 1353 (7.64%) and Giardia intestinalis 348 (1.96%) in stool samples, with Enterobius vermicularis 253 (4.25%) in cellophane tape preparations. Two or more parasites were detected in 158 (6.76%) of the positive cases. The mean age of persons identified as having a parasite was 21.9. Parasite incidence was 10.7% in females, and 13.6% in males (p < 0.001). The highest ratio was 23.8% in patients who lived in rural areas when all positive cases were evaluated in terms of settlement areas. CONCLUSION: While Blastocystis spp., E. vermicularis and G. intestinalis were the most frequently detected intestinal parasites in our province, intestinal parasites still remain important despite advances in infrastructure in recent years.