[Investigation of potentially pathogenic free-living amoebae and their in vivo pathogenicity in water supplies of Turkey]. / Türkiye'deki sularda serbest yasayan potansiyel patojen amipler ve patojenitelerinin in vivo olarak arastirilmasi.

Free-living amoebae (FLA) are found widely in soil and water in the nature. Among them in which potentially pathogenic for humans and animals are known as "potential pathogenic free-living amoebae (PPFLA)". PPFLA are characterized as the causes of clinical manifestations leading to death especially in immunosuppressed people. Four genus of PPFLA (Acanthamoeba, Naegleria, Balamuthia and Sappinia) are known to be pathogenic to humans. The aims of this study were to investigate the presence of PPFLA in the water supplies in Turkey and to determine their in vivo pathogenicity. A total of 664 water samples were collected from the ponds, rivers, streams and wells found in provinces located at different regions (central, western, eastern and southeastern regions) of Turkey. These samples were initially inoculated in the monoxenic culture media and evaluated by both microscopy and polymerase chain reaction (PCR) in terms of the presence of FLA. The samples identified as positive were then cultured in axenic media, the growth of amoebae that were confirmed microscopically, were than studied with PCR for molecular characterization. The isolates that were found positive by PCR from axenic cultures were inoculated intranasally to immunocompetent and immunodeficient (athymic) [BALB/c Rag2(-/-) gamma(c)(-/-)] BALB/c mice followed by the evaluation on the 21st day by histopathological and molecular methods to investigate their in vivo pathogenicity. In our study, 143 water samples were detected as positive in monoxenic cultures and 41 of them were detected as positive in axenic cultures. Twenty of 41 samples detected as positive in axenic culture could be continued in culture for three months. As a result of PCR using primers common to SYA, only nine have been identified from 20 samples as positive. According to the result of the PCR with specific primers, all (n= 9) were positive for Acanthamoeba sp., eight for Sappini sp. and five for Balamuthia mandrillaris, while none was observed Naegleria fowleri. Histopathologic examination revealed that both groups of mice that were infected with the nine isolates had normal brain tissue sections; but haemorrhages and mononuclear cell proliferation were determined in four immunocompetent and seven athymic animal lung sections. When the presence of parasites in tissue samples were evaluated by real-time PCR, Balamuthia was detected in at least one blood, lung, brain or nasal mucosa sample of the four immunocompetent mice, Sappinia sp. in four and Acanthamoeba sp. in seven immunocompetent mice infected with nine isolates. Additionally, seven Balamuthia sp., seven Sappinia sp. and eight Acanthamoeba sp. were detected in immunodeficient mice. In this study, B. mandrillaris and Sappinia sp. were the first isolated potentially pathogenic amoebae from water supplies located at different parts of Turkey. As a result awareness and precautions against suspicious water supplies used for drinking, daily use and swimming purposes should be treated more carefully.

[Investigation of the presence of Encephalitozoon intestinalis and Enterocytozoon bieneusi in bone marrow transplant patients by IFA-MAbs method]. / Kemik iligi transplant hastalarinda Encephalitozoon intestinalis ve Enterocytozoon bieneusi varliginin IFA-MAbs yöntemiyle arastirilmasi.

Microsporidian pathogens are obligatory intracellular eukaryotic parasites which can be found worldwide. They have been represented in 144 genera and more than 1200 species that may cause infections in both vertebrate and invertebrate hosts. Enterocytozoon bieneusi and Encephalitozoon intestinalis are the most common species among 14 species of microsporidia identified as human pathogens and they cause infections in the gastrointestinal tract. These species may also cause chronic diarrhea particularly in immunocompromised patients, as well as disseminated infections with severe clinical conditions which can be life-threatening. Since the spores of microsporidia are quite small-sized structures, they frequently may be overlooked in routine stool examinations. Therefore, molecular methods and transmission electron microscopy, if possible, are used as the gold standard methods in laboratory diagnosis. In laboratories in which those methods could not be applied, immunofluorescence assay using monoclonal antibodies (IFA-MAbs) may be advantageous compared to conventional methods. The aim of this study was to investigate the presence of E.intestinalis and E.bieneusi in bone marrow transplant (BMT) patients by using IFA-MAbs method. A total of 200 BMT patients (134 male, 66 female; mean age: 43.2±15.01 years), of them 147 with diarrhea and 80 healthy subjects (43 male, 37 female; mean age: 31.9±11.76 years) as control group were included in the study. All of the stool samples were examined by a commercial IFA-MAbs (Bordier Affinity Products, Switzerland) method as well as conventional (native-lugol and modified acid-fast staining) methods. Of the patients 25.5% (51/200) were positive for E.intestinalis, 4% (8/200) for E.bieneusi and 9.5% (19/200) for both of them, giving a total positivity rate of 39% (78/200). Those rates were 5% (4/80), 2.5% (2/80), 3.8% (3/80) and 11.3% (9/80), respectively for control group. The difference between the patient and control groups in terms of positivity was found statistically significant (39% vs 11.3%, p<0.05). Among 78 positive BMT patients, 67 (85.9%) were suffering from diarrhea. The correlation between the presence of diarrhea and the presence of microsporidia was statistically significant (p<0.05). It was concluded that, BMT patients particularly those with gastrointestinal complaints, have to be evaluated for microsporidian pathogens regularly to improve quality of life and to decrease the problems during the treatment period.

[Prevalence of Encephalitozoon intestinalis and Enterocytozoon bieneusi in cancer patients under chemotherapy]. / Kemoterapi alan kanserli hastalarda Encephalitozoon intestinalis ve Enterocytozoon bieneusi prevalansi

Microsporidia species are obligate intracellular parasites and constitute one of the most important opportunistic pathogens that can cause severe infections especially in immunocompromised patients. Enterocytozoon bieneusi and Encephalitozoon intestinalis are the most common species among 14 microsporidia species identified as human pathogens. The aim of this study was to investigate the prevalence of E.intestinalis and E.bieneusi in cancer patients under chemotherapy by immunofluorescent antibody and conventional staining methods. A total of 123 stool samples obtained from 93 patients (58 male, 35 female) with cancer who were followed in oncology and hematology clinics of our hospital and 30 healthy volunteers (13 male, 17 female) were included in the study. Fifty-one (55%) of the patients had complain of diarrhea. The presence of E.intestinalis and E.bieneusi were investigated by a commercial immunofluorescence antibody test using monoclonal antibodies (IFA-MAbs; Bordier Affinity Products, Switzerland) in all of the samples, and 50 of the samples were also investigated by modified trichrome, acid-fast trichrome and calcofluor staining methods. A total of 65 (69.9%) patients were found positive with IFA-MAbs method, including 43 (46.2%) E.intestinalis, 9 (9.7%) E.bieneusi and 13 (14%) mixed infections. In the control group, 5 (16.7%) subjects were positive with IFA-MAbs method, including 2 (6.7%) E.intestinalis, 1 (3.3%) E.bieneusi and 2 (6.7%) mixed infections. The difference between the positivity rate of the patient and control groups was statistically significant (p< 0.05). Of the patients with diarrhea, 68.6% (35/51) were infected with microsporidia, and the difference between cases with and without (48.6%) diarrhea was statistically significant (p< 0.05). When 50 samples in which all of the methods could be performed were evaluated, the frequency of microsporidia were detected as follows; 66% (n= 33) with IFA-MAbs, 34% (n= 17) with modified trichrome staining, 24% (n= 12) with acid-fast trichrome staining and 42% (n= 21) with calcofluor staining methods. Our data indicated that the use of IFA-MAbs method along with the conventional staining methods in diagnosis of microsporidia will increase the sensitivity. As a conclusion, the prevalence of E.intestinalis and E.bieneusi in cancer patients under chemotherapy was detected quite high (69.9%) in our study, it would be appropriate to screen these patients regularly in terms of microsporidian pathogens.

[Evaluation of imported Plasmodium falciparum malaria cases: the use of polymerase chain reaction in diagnosis]. / Yurt Disi Kaynakli Plasmodium falciparum Sitmali Olgularin Degerlendirilmesi: Tanida Polimeraz Zincir Reaksiyonunun Yeri.

Malaria affecting almost half of the world population continues to be an important health problem. Although domestic malaria cases have been decreasing in Turkey recently, cases caused by Plasmodium falciparum have increased due to the frequent travelling to Africa. The aims of this study were to evaluate demographic characteristics, clinical and laboratory findings in cases with falciparum malaria who attended to our clinic in 2012-2013 period, and the impact of polymerase chain reaction (PCR) for diagnosis. Nine patients evaluated were all male with a mean age of 34.3 (age range: 18-48) years, with the history of travel to Africa. Six cases did not take prophylaxis against malaria and other three cases used insufficient time. Mean duration of symptoms after return was 18.4 (range: 1-75) days, and the patients were admitted to the clinic within a mean of 5.2 (range: 1-15) days. Two patients had leucopenia, two patients had anemia, and eight patients had thrombocytopenia on admission. Alanine aminotransferase (ALT) levels in four cases and total bilirubin levels of six cases were over upper normal limits. Definitive diagnosis of cases was performed with the detection of ring and/or gametocytes forms of the parasite in Giemsa-stained peripheral blood smears. Furthermore, samples from seven patients were studied by nested PCR by using genus (Plasmodium rPLU 1 and 5) and species (rFAL 1 and 2, rVIV 1 and 2, rMAL 1 and 2, rOVA 1 and 2) specific primers. All of these seven samples yielded positive results with primers specific for P.falciparum ssrRNA. In the treatment, arthemeter/lumefantrin and doxycycline combination was used in seven patients, while intravenous artesunate and doxycycline combination was given to two patients, resulting with complete cure. Mean duration for the resolving of fever was 3.3 days, and mean duration for clearing the parasitemia from peripheral blood was 4.9 days. Initial ALT values and the duration of fever resolution (-796; p= 0.010), as well as the duration of parasitemia and initial thrombocyte counts (-797; p= 0.010) were negatively- correlated. It was concluded that, providing sufficient information on malaria and prophylaxis to people travelling to the endemic areas are crutial for protection. Moreover, in endemic areas for Crimean-Congo hemorrhagic fever, patients with fever and thrombocytopenia should be questioned in detail about the travel history, and peripheral blood smears should be examined in terms of malaria, since their clinical features are similar. Plasmodium PCR should be considered as one of the alternative diagnostic method in malaria, especially in cases with inconclusive microscopy.

Encephalitozoon intestinalis: A Rare Cause of Diarrhea in an Allogeneic Hematopoietic Stem Cell Transplantation (HSCT) Recipient Complicated by Albendazole-Related Hepatotoxicity.

UNLABELLED: A 50-year-old male patient previously diagnosed with acute myelomonocytic (M4) leukemia in July 2009 underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT). During the pre-transplant period complete blood count (CBC), liver and renal function tests, coagulation tests, and other parameters were normal. On the first day of transplantation teicoplanin (400 mg d-1 for the first 3 d, and then 400 mg d-1) and caspofungin (first dose was 1×70 mg d-1, followed by 1×50 mg d-1) were started intravenously due to white plaques and oropharyngeal candidiasis in the patient's mouth and perianal erythema. On the 14th d of transplantation watery diarrhea occurred, along with abdominal discomfort, nausea, and fatigue. Stool examination was negative for findings of bleeding. Investigation of Microsporidia confirmed a rare pathogen Encephalitozoon intestinalis in the patient's stool sample via species-specific immunofluorescence antibody (IFA) assay and albendazole treatment was started at a dose of 2×400 mg d-1. On the 5th d of albendazole treatment (d 18 of treatment) liver function test (LFT) results began to deteriorate. As LFT results continued to deteriorate, albendazole was withdrawn on the 7th d of treatment. Biopsy was performed on the 22nd d of transplantation and histopathological analysis confirmed the diagnosis of toxic hepatitis. LFT results began to decrease after withdrawal of albendazole treatment. On the 13th d of albendazole treatment all LFT values returned to normal. The presented allo-HSCT case had a rare pathogenic agent (E. intestinalis) that caused diarrhea, as well as hepatotoxicity due to albendazole treatment. This is the first reported case of E. intestinalis diagnosed via IFA in Turkey. CONFLICT OF INTEREST: None declared.

Stool sample storage conditions for the preservation of Giardia intestinalis DNA.

Stool is chemically complex and the extraction of DNA from stool samples is extremely difficult. Haemoglobin breakdown products, such as bilirubin, bile acids and mineral ions, that are present in the stool samples, can inhibit DNA amplification and cause molecular assays to produce false-negative results. Therefore, stool storage conditions are highly important for the diagnosis of intestinal parasites and other microorganisms through molecular approaches. In the current study, stool samples that were positive for Giardia intestinalis were collected from five different patients. Each sample was stored using one out of six different storage conditions [room temperature (RT), +4ºC, -20ºC, 70% alcohol, 10% formaldehyde or 2.5% potassium dichromate] for DNA extraction procedures at one, two, three and four weeks. A modified QIAamp Stool Mini Kit procedure was used to isolate the DNA from stored samples. After DNA isolation, polymerase chain reaction (PCR) amplification was performed using primers that target the ß-giardin gene. A G. intestinalis-specific 384 bp band was obtained from all of the cyst-containing stool samples that were stored at RT, +4ºC and -20ºC and in 70% alcohol and 2.5% potassium dichromate; however, this band was not produced by samples that had been stored in 10% formaldehyde. Moreover, for the stool samples containing trophozoites, the same G. intestinalis-specific band was only obtained from the samples that were stored in 2.5% potassium dichromate for up to one month. As a result, it appears evident that the most suitable storage condition for stool samples to permit the isolation of G. intestinalis DNA is in 2.5% potassium dichromate; under these conditions, stool samples may be stored for one month.

Oxidative stress in vivax malaria.

Malaria is still a leading cause of morbidity and mortality. The increase in lipid peroxidation reported in malaria infection and antioxidant status may be a useful marker of oxidative stress during malaria infection. The aim of this study was to investigate the role of antioxidant enzymes against toxic reactive oxygen species in patients infected with Plasmodium vivax and healthy controls. Malondialdehyde levels, superoxide dismutase, and glutathione peroxidase activities were determined in 91 P. vivax patients and compared with 52 controls. Malondialdehyde levels, superoxide dismutase, and glutathione peroxidase activities were 8.07±2.29 nM/ml, 2.69±0.33 U/ml, and 49.6±3.2 U/g Hb in the patient group and 2.72±0.50 nM/ml, 3.71±0.47 U/ml, and 62.3±4.3 U/g Hb in the control group, respectively. Malondialdehyde levels were found statistically significant in patients with vivax malaria higher than in healthy controls (P<0.001). On the other hand, superoxide dismutase and glutathione peroxidase activities were found to be significantly lower in vivax malaria patients than in controls (P<0.05). There was an increase in oxidative stress in vivax malaria. The results suggested that antioxidant defense mechanisms may play an important role in the pathogenesis of P. vivax.

Could Toxoplasma gondii have any role in Alzheimer disease?

BACKGROUND: Alzheimer disease (AD), a chronic progressive neurodegenerative disorder, has a mainly unknown multifactorial etiology. Neuroinflammatory mechanisms might contribute to the cascade of events leading to neuronal degeneration. Central nervous system infections have been previously suggested as possible etiological agents in the development of sporadic AD. Toxoplasmosis can be associated with various neuropsychiatric disorders. In this study, we aimed to investigate the possible association between toxoplasma infection and AD. METHODS: This study evaluated the serum anti-Toxoplasma gondii IgG levels. It included an age-matched and sex-matched study and control groups that consisted of 34 patients with AD and 37 healthy individuals, respectively. There were no difference between the socio economic states of the patients and control subjects. serecm anti-I-gondi IgG levels were measured by using ELISA. RESULTS: According to the statistical analysis, there were no significant differences among the patients and the control participants with respect to age (68.05±15.98, 62.91±5.89 y, P=0.072; respectively) and sex. The seropositivity rate for anti-T. gondii IgG antibodies among AD patients and control groups were 44.1% and 24.3%, respectively, and there was significant difference between the serum anti-T. gondii IgG levels (P=0.005). CONCLUSIONS: Our findings suggest that toxoplasma infection may be involved in the pathogenetic mechanisms of AD. If confirmed, a positive correlation between toxoplasmosis and AD may lead to new approaches for the management of AD.

Is Toxoplasma gondii a causal agent in migraine?

BACKGROUND: Many different tissues may be parasitized by Toxoplasma gondii, particularly, lung, heart, lymphoid organs, and the central nervous tissues. Tissue cysts of this parasite in the brain may spontaneously rupture, releasing parasites that cause antibody titers to rise. In immunocompetent subjects with acquired toxoplasmosis, the most frequent symptoms were lymphadenopathy and headache. In the neurogenic inflammation theory of the pathogenesis of migraine, the cause of initial release of ions and inflammatory agents has not been established. In this study, we aimed to investigate if T. gondii infection is a possible cause of neurogenic inflammation of migraines. METHODS: The anti-T. gondii antibody status of 104 patients with migraine were studied and compared with those of control groups, 50 healthy subjects and 50 subjects with headache due to rhinosinusitis, by using a micro-enzyme-linked immunosorbent assay technique. RESULTS: Forty-six (44.2%) patients with migraine, 13 (26.0%) healthy control subjects, and 12 (24%) control subjects with rhinosinusitis were positive for anti-T. gondii IgG antibody. The rate of positivity in the migraine patient group was statistically different from those of the control groups (P < 0.05). CONCLUSIONS: The results show the presence of chronic Toxoplasma infection in patients with migraine. Toxoplasma infection may contribute to neurogenic inflammation as the pathogenesis of migraine, as many studies in the literature have reported that Toxoplasma infection causes biochemical and immunologic changes.

The schizophrenia and Toxoplasma gondii connection: infectious, immune or both?

INTRODUCTION: Recent research has suggested a possible link between toxoplasmic agents and schizophrenia. We aimed to assess this by measuring Toxoplasma gondii-associated antibodies in schizophrenia patients and controls METHODS: We used a commercially available enzyme-linked immunosorbent assay (ELISA) kit to measure the level of immunoglobulin G (IgG) and IgM antibodies in serum samples from schizophrenia patients (n=40) and from a group of non-schizophrenic control subjects (n=37) RESULTS: Among schizophrenic patients, 16 (40%) showed IgG seropositivity and two (5%) showed IgM seropositivity. Among the control group, five (13.5%) were found have IgG seropositivity and one (2.7%) showed IgM seropositivity. In our study we found that IgG T gondii antibodies were significantly higher in schizophrenia patients compared with controls CONCLUSIONS: This study supports the theory that toxoplasmic agents may have a role in the aetiology of schizophrenia.

Anti-Toxoplasma gondii antibodies in patients with schizophrenia--preliminary findings in a Turkish sample.

Schizophrenia is a serious neuropsychiatric disease of uncertain etiology. We investigated the seropositivity rate for anti-Toxoplasma IgG and IgM antibodies by enzyme-linked immunosorbent assay (ELISA) in patients with schizophrenia to ascertain a possible relationship between Toxoplasma gondii and schizophrenia. We selected 100 patients with schizophrenia, 50 with depressive disorder, and 50 healthy volunteers to investigate the seropositivity rate of anti-Toxoplasma antibodies by ELISA. The seropositivity rate for anti-Toxoplasma IgG antibodies among schizophrenia patients (66%) was significantly higher than among patients with depressive disorder or healthy volunteers (P < .01). Thus, there might be a causal relationship between toxoplasmosis and the etiology of schizophrenia.

A Case Series of Imported Malaria Caused by Plasmodium falciparum in Kayseri and Review of Literature.

Malaria is a major worldwide public health problem. In the last years, no domestic cases of malaria have been detected and cases of imported malaria exist only in Turkey. In this study, clinical and laboratory findings of five Plasmodium falciparum (P. falciparum) malaria patients who were admitted to the emergency department between January 2013 and December 2015 were retrospectively presented. One of the patients was an African student, and the other patients had a history of travelling to Africa. Ring formation was observed when Giemsa staining was performed on the blood smears of all patients, and in three patients, P. falciparum was also detected using multiplex polymerase chain reaction (PCR) (Bio-Rad, United States of America). P. falciparum was not detected by PCR in the other two patients. Malaria should be primarily considered in febrile patients who have a history of travelling to endemic regions, and peripheral blood smears should definitely be examined.

IgG and IgG subclass antibodies in patients with active cutaneous leishmaniasis.

This research was planned to detect IgG and IgG subclasses in sera of patients with active cutaneous leishmaniasis (CL). Sera from 30 patients with active CL aged between 10 and 50 years and from 30 healthy controls aged between 8 and 50 years were included in the study. Levels of IgG and its subclasses were measured by a nephelometer. Levels of IgG, IgG1 and IgG3 in the CL patients were higher than in the controls. In addition, IgG and IgG1, and IgG and IgG3 levels showed a significant positive correlation. These results showed that IgG subclasses could possibly be used as a helpful diagnostic marker in CL.

Anti-Toxoplasma gondii antibodies in patients with chronic heart failure.

Chronic heart failure (CHF) involves interactions between the cardiovascular, neuroendocrine and immune systems. This study investigated the seropositivity rate for anti-Toxoplasma IgG and IgM antibodies by ELISA in patients with CHF. Ninety-seven patients with CHF and 50 healthy volunteers were selected for this investigation. The seropositivity rate for anti-Toxoplasma IgG antibodies among CHF patients (68%) was significantly higher than in healthy volunteers (36%). Thus, parasitological screening of this group of patients should be periodically performed to prevent the possible dissemination of toxoplasmosis.

Teniasis: effect on element status of children.

The aim of this study was to investigate the changes of level of the essential elements of copper, magnesium, and zinc status in cases of teniasis in children. Copper, magnesium, and zinc levels were measured in 40 children who were positive for intestinal parasite of Taenia saginata. Scores were obtained for the positives and their 30 age- and sex-matched T. saginata-negative healthy children. The mean concentration of copper, magnesium, and zinc in blood showed no statistically difference in T. saginata-positive children than in their controls both in females (p>0.05) and males (p>0.05). However, a clear numerically decrease was observed especially in magnesium and zinc levels compared to the controls both in females and males. The average magnesium concentration in T. saginata-positive female children and male children were 20+/-1.9 and 22+/-2.2 mg/L and it was 27+/-2.1 and 27+/-2.3 mg/L in controls, respectively. The mean values of the zinc in blood were 0.76+/-0.5 and 0.72+/-0.4 mg/L in T. saginata-positive female children and male children and 0.85+/-0.3 and 0.81+/-0.5 mg/L in female and male controls, respectively. No correlation could be demonstrated between age and mean values of copper, magnesium, and zinc in T. saginata-positive females and males and controls (p>0.05). No significant correlation could be found between blood copper, magnesium and zinc levels in T. saginata-positive female and male children and controls (p>0.05). Although there was no statistical correlation observed in copper, magnesium, and zinc levels between patients and controls, there seem to be, especially in magnesium and zinc levels, a decrease, whereas no change was seen in the zinc level in children infected with T. saginata compared to controls.

Dust-mites: effect on lipid peroxidation.

BACKGROUND: Dust-mites are present in our homes, feed on dead exfoliated skin and other organic material. It is also known that oxidative stress may lead to cellular damage that can be confirmed by markers of cellular disruption. Oxidative stress in various infective processes has been documented. We investigated whether house dust-mites cause oxidative stress in patients. METHODS: Products of lipid peroxidation in erythrocytes and lymphocytes were assessed by measuring malondialdehyde concentration. RESULTS: Our results showed that patients who had a positive skin test for dust-mite antigens and had dust-mites present in their houses (dust-mite positive) had increased erythrocyte malondialdehyde levels (62.39 [18.56] nmol/g-Hb) compared with those who were skin test positive, dust-mite negative (45.45 [10.82]) or skin test negative, dust-mite negative (42.20 [5.68]). They also had significantly higher levels of lymphocyte malondialdehyde (4.22 [0.55] nmol/g-protein) compared with those who were skin test positive, dust-mite negative (3.46 [0.29]) or skin test negative, dust-mite negative (1.25 [0.31]; p <0.05). However, there was no statistically significant difference between the malondialdehyde levels of dust-mite negative/skin test positive and dust-mite negative/skin test negative patients. CONCLUSION: Increased malondialdehyde activity in lymphocytes and erythrocytes in the dust-mite positive/skin test positive group shows the presence of the oxidative stress in patients with dust-mite infestation.

Prevalence of anti-toxoplasma Gondii antibodies in Turkish blood donors.

OBJECTIVE: Toxoplasmosis is caused by an obligate intracellular protozoan parasite, Toxoplasma gondii. Most of these infections are asymptomatic or benign, but may cause severe or fatal consequences in immunodeficient patients, transplant recipients, and in the fetus. Transmission may occur by eating uncooked meat, contaminated vegetables, blood transfusion, organ transplantation, and across the placenta from the mother to the fetus. IgG antibodies to T. gondii may persist in the serum at high titers for years. In the present study, our aim was to determine prevalence of anti- T. gondii IgG and IgM antibodies in Turkish blood donors. METHODS: A total samples from 385 healthy blood donors from Kayseri, Turkey were examined for anti- T. gondii antibodies by indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA). RESULTS: The seroprevalence of the anti- T. gondii IgG and IgM antibodies was 19.5%, 2.33% respectively by IFAT and 20.25%, 2.33% by ELISA. CONCLUSION: It is suggested that all blood donors should be screened for toxoplasmosis before transfusion

Cystic echinococcosis in Central Anatolia, Turkey.

OBJECTIVE: Human cystic echinococcosis (CE) caused by infection with a larval stage of Echinococcus granulosus is a serious public health problem in Turkey. Echinococcosis is a zoonotic disease; dogs and livestock are important hosts in transmission. The aim of this study is to evaluate the rate of CE in Kayseri Rural Area, Central Anatolia, Turkey. METHODS: At the present study, we planned to evaluate the rate of CE in Kayseri rural area in Central Anatolia between 2000 and 2002. We investigated 2,242 subjects using enzyme-linked immunosorbent assay (ELISA) and indirect fluorescence antibody (IFA), and we examined the seropositivity by using Western blotting (WB). RESULTS: The seropositivity rate was 2.7% by ELISA and IFA. We retested seropositive serum samples and 200 seronegative sera by WB. Seropositive serum samples were studied using abdominal ultrasound and chest x-ray to confirmed the presence of hydatid cyst and we found 10 (0.5%) different localized cysts. CONCLUSION: The results of our study indicate that Kayseri rural area has a high endemicity of human CE.

Cloning of the DNA Vaccine Candidate LACK Gene of Leishmania infantum.

OBJECTIVE: Leishmaniasis is caused by an obligate intracellular protozoa belonging to Leishmania genus and listed among major tropical diseases by WHO. Because of the high costs, toxicity, and adverse effects of routinely used compounds in the treatment, alternative treatment and vaccine studies are underway. An effective vaccine has not been developed to date. In this study, we aimed to clone one of the most promising DNA vaccine candidates: the homolog-activated C kinase (LACK) gene of Leishmania infantum. METHODS: L. infantum genomic DNA was isolated from promastigote culture. The LACK gene was placed into plasmid pJET1.2. Then, recombinant plasmids were transformed into competent cells. The presence of recombinant plasmids was determined by PCR screening. Cloning was confirmed by PCR, restriction enzyme assays, and finally, DNA sequence analysis, after making miniprep from positive colonies. RESULTS: After performing PCR with LACK-gene specific primers, 939-bp PCR products were observed. Recombinant plasmids, which were transformed into competent Escherichia coli cells, were verified by PCR screening. It was verified by PCR that the recombinant plasmid contained the LACK gene. DNA sequence analysis was performed to obtain the DNA sequence. CONCLUSION: One of the most promising DNA vaccine candidates against leishmaniasis, the LACK gene, was cloned in this study.

Microbiota: In Health and in Sickness, From Birth to Death.

Microorganisms colonize tissues and organs such as the skin and gastrointestinal, respiratory, and genitourinary systems. These microorganisms are generally called as "human microbiota". Human microbiota mostly consists of commensal microorganisms. The commensal microorganisms located on and in the human body are bacteria, fungi, viruses, archaea, and parasites. The microbiota genome is 100 times bigger in size than the human genome. Although the human genome is stationary, microbial genome has a compatible flexible variability during human life. As well as 2-year-old child and newborn, adult and adolescent, the elderly and pregnant woman have a different microbiota. Microbiota and the microbiota genome can be changed by personal and household diet, antibiotic use, mode of delivery, and hygiene within days or even hours, depending on such as these factors. The human immune system and microbiota grow up, develop, and mature as childhood friends by playing with each other from birth to death. Association between microbiota and human is not just related to childhood-it continues with health and disease, until death separates them. This review focused on the roles of microbiota in parasitology, autoimmune diseases, metabolic diseases, and cancer treatment in detail. In addition, inflammatory and immunoregulatory roles of microbiota on the intestinal immune system and how innate and adaptive immune systems regulate microbiota and its content were explained.