[Method Verification of Inhouse Real-time Polymerase Chain Reaction for Detection of Toxoplasma gondii]. / Toxoplasma gondii'nin Inhouse Gerçek Zamanli Polimeraz Zincir Reaksiyonu ile Saptanmasinda Yöntemin Dogrulanmasi.

Toxoplasmosis is one of the most common parasitic infection in humans. Serological and molecular methods are used for diagnosis. Molecular methods are becoming increasingly preferred, since they lead to shortening of diagnostic time. In our study, it was aimed to determine Toxoplasma gondii by a cost-effective, quantitative, fast and reliable method without using a commercial kit, and apply method verification. T.gondii strain which was continued by mouse inoculation in our laboratory was used for method verification study. For this purpose DNA extraction was performed using a commercial kit. The limit of detection and, high and low positivity rates were determined by serial dilutions of DNA sample. Accuracy and certainty studies were performed using with TG-F, TG-R primers and TaqMan TG probe for method verification of the test. In the study with serial dilutions of DNA sample, detection limit was determined as 10-3 dilutions (0.028 copies/reaction). Furthermore 10-1 dilution (2.8 copies/reaction) was considered as high positive, 10-2 dilution (0.28 copies/reaction) was considered as low positive and method verification studies were performed. The accuracy of test was determined as 0.62 for high positive samples and 0.14 for low positive samples. CV value of intra-assay certainty was 0.62 for high positive samples and 0.14 for low positive samples, whereas, CV value of inter-assay certainty was calculated as 1.03 for high positive samples and 2.34 for low positive samples. Correlation coefficient was determined as 0.99. The coefficient of variation of inhouse realtime PCR method used in our study was found to be below 15%, and it was decided to be suitable for routine laboratory studies.

[Cutaneous larva migrans in Turkey: an imported case report]. / Türkiye'de kütanöz larva migrans: bir importe olgu sunumu.

Cutaneous larva migrans (CLM) is a parasitic infection most commonly found in tropical and subtropical areas. However, with the ease and increase of foreign travel to many countries around the world, the infection is not limited to these areas. CLM is an erythematous, serpiginous infection with skin eruption caused by percutaneous penetration of the larvae to the skin. In this report, a case diagnosed as imported CLM after an Amazon trip and treated with albendazole was presented. A 36 year-old male patient admitted to infectious diseases clinic with intense itching, erythematous, raised, streaklike serpiginious eruptionand some redness at bilateral foot especially at the right foot for about one week. The patient was living in Turkey, and travelled to Brazil for an Amazon trip three months ago and the lesions began immediately after this occasion. CLM was diagnosed with the typical lesions in the patient and oral albendazole treatment 2 x 400 mg/day for 3 consecutive days was carried out with oral amoxicillin/clavulanat 3 x 1 g/day for the secondary bacterial infection. The patient responded very well to oral albendazole treatment with a result of a rapid improvementof pruritus in days and no side effect was observed during the treatment period.After discharge, during his controlit was seenthat the lesions were regressed with leaving hyperpigmentation. In cases with cutaneous larva migrans, diagnosis is often made by the presence of pruritic typical lesions and tunnels, travel story to endemic regions, the story of barefoot contact with sand and soil in these regions, and the sun tanning story on the beach. The lesions are often seen in the lower extremities, especially in the dorsal and plantar surface of the foot. Laboratory findings are not specific. Temporary peripheral eosinophilia can be seen and biopsy can be done to confirm the diagnosis but usually no parasite is seen in the histopathological examination. Contact dermatitis, bacterial and fungal skin infections and other parasitic diseases should be considered in differential diagnosis. For the treatment ivermectin 1 x 200 mg/kg single dose or albendazole 400 mg/day for three days is recommended. As a result, cutaneous larva migrans should be kept in mind especially in patients with a history of travel to endemic areas and a history of bare feet contact with sandy beaches and soil in this region and with itchy, red and serpiginous skin lesions.

Dobrava-Belgrade virus in Apodemus flavicollis and A. uralensis mice, Turkey.

In 2009, human Dobrava-Belgrade virus (DOBV) infections were reported on the Black Sea coast of Turkey. Serologic and molecular studies of potential rodent reservoirs demonstrated DOBV infections in Apodemus flavicollis and A. uralensis mice. Phylogenetic analysis of DOBV strains showed their similarity to A. flavicollis mice-borne DOBV in Greece, Slovenia, and Slovakia.

Investigation of the effects of Toxoplasma gondii on behavioral and molecular mechanism in bradyzoite stage.

Toxoplasma gondii is a single-celled parasite that causes a disease called toxoplasmosis. It can reach the central nervous system, but the mechanism of T. gondii disrupting the functioning of these brain regions occurs in bradyzoite stage of parasite, causing brain damage by forming tissue cysts in brain. In our study, the effects of T. gondii on locomotor activity, anxiety, learning and memory, and norepinephrine (NE), levodopa (L-DOPA), dopamine (DA) and 3,4-D-dihydroxyphenylacetic acid (DOPAC) catecholamines in amygdala, striatum, prefrontal cortex and hippocampus regions of the brain were investigated in bradyzoite stage. Twenty male Albino mice Mus musculus, 4-5 weeks old, weighing 20-25 g, were used. T. gondii inoculated to mice intraperitonealy with 48-50-hour passages of T. gondii RH Ankara strain. For intraperitoneal inoculation of mice 5x104 tachyzoites per mouse. No inoculation was made in control group (n: 20). Locomotor activity behavior in open field test (OFT), anxious behavior in elevated plus maze (EPM), and learning behavior in novel object recognition (NOR) tests were evaluated. NE, L-DOPA, DA and DOPAC were measured by HPLC in brain tissues of amygdala, striatum, prefrontal cortex and hippocampus. A decrease was observed in the locomotor activity, anxiety and learning values of the T. gondii group compared to the control group (p < 0.05). The heighten in NE and L-DOPA levels in amygdala tissue of T. gondii group compared to control group, an elevation in NE, L-DOPA, DA and DOPAC levels in striatum tissue, and an increase in levels of NE in prefrontal cortex tissue were detected in monoamine results. In hippocampus tissue, an increase was observed in DA levels, while a decrease was observed in NE, L-DOPA and DOPAC levels. In our study, it has been shown that T. gondii in bradyzoite stage reduces locomotor activity, causes learning and memory impairment, and has anxiogenic effects.

Investigation of resistance against to flumethrin using against Varroa destructor in Türkiye.

The honeybee ectoparasite Varroa destructor is a major threat to apiculture when evaluating bee diseases and pests. While attempting to control this mite, beekeepers often depend on a small selection of authorized synthetic acaricides, such as flumethrin, which is widely used in Türkiye and globally. However, resistance to flumethrin develops due to incorrect and excessive use. In this study conducted at Ordu Beekeeping Research Institute, trial group were established including an untreated control group and group where flumethrin-based pesticides were applied. Dead varroas collected from pollen traps and live varroas collected from bees were obtained from these trial groups for molecular analysis as positive-negative controls. Varroa samples were collected from provinces representing different regions with intensive beekeeping activities such as Adana, Ankara, Bingöl, Mugla, Ordu, Sanliurfa, Tekirdag. Molecular methods were employed to investigate the resistance gene region for pyrethroids (specifically flumethrin) against V. destructor. In our study, individual DNA extractions were performed on dead parasites from colonies subjected to pyrethroid application (resistance negative control) and live parasites (resistance positive control). The DNA samples obtained were used in PCR reactions targeting the region encoding the 925th amino acid of the voltage-gated sodium channel (VGSC) gene, which is responsible for resistance formation. The DNA samples were subjected to gel electrophoresis to observe the amplification products of the expected target region. To examine the nucleotide sequence changes that encode leucine at the 925th amino acid, which is associated with resistance, DNA sequence analysis was applied to the amplification products. Out of 332 V. destructor parasites obtained from different provinces, 279 were analysed using molecular methods. It was observed that 31% of the samples showed sensitivity to flumethrin while 69% exhibited resistance to it. Among the resistant samples: 27% had homozygous isoleucine mutation; 28% had homozygous valine mutation; 2.8% had heterozygous isoleucine mutation; 8.5% had heterozygous valine mutation; and 2.8% had heterozygous methionine mutation, all of which were associated with flumethrin resistance. As a result, the rate of flumethrin resistance in parasites varied between 51% and 94% among different provinces.

An Overview of One Health Concept Focusing on Toxoplasmosis.

The "One Health" concept is a universal approach to sustainably balancing and optimizing the health of humans, animals, and ecosystems. This approach is based on the health of humans, domestic and wild animals, and plants in a wider environment in which self-renewable ecosystems exist, with essential characteristics of integration, unifying and holistic perspective. Toxoplasmosis, one of the most common zoonotic infections in both terrestrial and oceanic ecosystems in the world, is an ideal model disease for the "One Health" approach. Toxoplasmosis is a zoonotic disease caused by the obligate intracellular pathogen protozoan Toxoplasma gondii. In the life cycle of T. gondii, the definitive host is domestic cats and felines, and the intermediate hosts are all mammals (including humans), birds and reptiles. The infected cats have primary importance and play a crucial role in the contamination of habitats in the ecosystems with T. gondii oocysts. Thus, ecosystems with domestic cats and stray cats are contaminated with cat feces infected with T. gondii oocytes. T. gondii positivity has been scientifically demonstrated in all warm-blooded animals in terrestrial and aquatic habitats. The disease causes deaths and abortions in farm animals, resulting in great economic losses. However, the disease causes great problems in humans, especially pregnant women. During pregnancy, it may have effects such as congenital infections, lesions in the eye and brain of the fetus, premature birth, intrauterine growth retardation, fever, pneumonia, thrombocytopenia, ocular lesions, encephalitis, and abortion. The mechanism of death and abortion of the fetus in a pregnant woman infected with T. gondii occurs as a result of complete disruption of the maternal immune mechanism. The struggle against toxoplasmosis requires the universal collaboration and coordination of the World Organization for Animal Health, the World Health Organization and the World Food Organization in the "One Health" concept and integrative approaches of all responsible disciplines. Establishing universal environmental safety with the prevention and control of toxoplasmosis requires the annihilation of the feces of the infected cats using suitable techniques firstly. Then routinely, the monitoring and treatment of T. gondii positivity in cats, avoiding contact with contaminated foods and materials, and development of modern treatment and vaccine options. Particularly, mandatory monitoring or screening of T. gondii positivity during the pregnancy period in humans should be done. It would be beneficial to replace the French model, especially in the monitoring of disease in humans. In this article, the ecology of toxoplasmosis was reviewed at the base of the "One Health" concept.

Borrelia miyamotoi in wild rodents from four different regions of Turkey.

Borrelia miyamotoi is a tick-borne zoonotic agent that causes hard tick-borne relapsing fever, an emerging disease in humans. Some small mammalian and bird species are reported to be reservoirs of B. miyamotoi. This study aims to examine Borrelia species present in rodents captured from rural areas of Turkey. Blood samples of rodents were initially screened with Borrelia 16S rRNA qPCR. The Borrelia flaB gene was subsequently amplified by conventional PCR, after which all positive samples were sequenced. Borrelia miyamotoi was observed in nine out of 536 blood samples (1.7%) collected from wild rodents. Phylogenetic analysis showed that all positive samples belonged to the European genotype clade of B. miyamotoi. PCR positivity was 5.3%, 3.7%, and 1.8% in Apodemus uralensis, Apodemus flavicollis, and Myodes glareolus, respectively. Borrelia burgdorferi sensu lato that causes Lyme borreliosis in humans could not be detected in the rodents. In this study, presence of B. miyamotoi DNA is reported for the first time in rodents in Turkey.

Oxidative stress and tryptophan degradation pattern of acute Toxoplasma gondii infection in mice.

Toxoplasma gondii is a very common obligate single-cell protozoan parasite which induces overproduction of interferon (IFN)-gamma and of other proinflammatory cytokines. Although immunomodulatory role of IFN-gamma favors tryptophan (Trp) degradation via indoleamine-2,3-dioxygenase (IDO) activity and is related with nitric oxide (NO) synthesis, the mechanism of antitoxoplasma activity is complex. In order to characterize the Trp degradation pattern during the acute T. gondii infection, serum Trp, kynurenine (Kyn), and urinary biopterin levels of mice were measured. The possible oxidative status was evaluated by the liver, spleen, brain, and serum malondialdehyde (MDA) and NO levels. Increased free radical toxicity may cause elevation in tissue MDA in T. gondii-infected mice, while unchanged serum MDA might indicate the increased oxidative stress due to T. gondii infection restricted to intracellular area. Elevated serum NO most probably might be due to the formation of reactive nitrogen radicals. The Kyn/Trp ratio was higher in T. gondii-infected mice compared to healthy animals (p < 0.05); however, it was not correlated with urinary biopterin. These results suggested that Trp degradation might be promoted by a pathway other than IDO during T. gondii infection and the reduction of Trp concentration favors the local immunosuppression and systemic tolerance.

Evaluation of Echinococcosis Pre-Diagnosis Patients Admitted to the National Parasitology Reference Laboratory of Turkey from 2014-2019.

Background: Echinococcosis is a common parasite with zoonotic character created by a small cestode, Echinococcus spp., and is an important public health problem in Turkey as well as all over the world. We aimed to investigate antibodies in serum samples of suspected Echinococcosis patients sent to the National Parasitology Reference Laboratories of the General Directorate of Public Health. Methods: Serum samples of 2390 patients sent to our laboratory between January 1, 2014 and May 01, 2019, evaluated by ELISA, Indirect Hemagglutination Test (IHA) and Western Blot (WB) methods are presented. Our laboratory is the national reference laboratory. All kinds of tests requested from suspected patients can be performed. Results: Overall, 1199 (50.2%) of 2390 serum samples were female and 1191 (49.8%) were male. It was observed that 178 (14.9%) of men and 210 (17.5%) of women were seropositive. There was no statistical difference between the sexes in terms of seropositivity. Of all samples, 1941 (81.2%) were negative, 388 (16.2%) were positive, and 61 (2.6%) were borderline. Results determined as borderline are considered suspicious and a recommendation is made to repeat the test after 15 days. A statistical difference was found in the distribution of seropositivity by years. While seropositivity was lowest in 2014, it was found to be highest in 2018 and 2019. Conclusion: Despite all the precautions taken, it is seen that echinococcosis still continues to exist in Turkey as a zoonotic disease. Hence, CE has been involved in Turkey Zoonotic Diseases Action Plan (2019-2023) and decided to carry out studies for the protection and prevention of the disease.

[A case of urogenital myiasis caused by Psychoda albipennis (Diptera: Nematocera)]. / Psychoda albipennis'in (Diptera: Nematocera) neden oldugu ürogenital miyazis olgusu.

Myiasis which is a parasitic disease of humans and vertebrates, is caused by dipterous fly larvae feeding on the host's necrotic or living tissue. Although infestation by fly larvae is much more prevalent in animals, it occurs relatively frequently in humans in rural, tropical and subtropical regions of Africa and America. Myiasis is usually associated with poor general health and hygiene, restricted mobility and ulcerating lesions. The pathophysiology of the human infection differs depending on the fly species and where the larvae are located. It could be external or internal, and the invasion by the maggot could be obligatory, facultative and sometimes acci-dental. Myiasis is a self-limiting infestation with minimal morbidity in the vast majority of cases. Urogenital myiasis, associated with urinary obstruction, poor hygiene of the local site and ulcerating lesions has been infrequently reported. In this report, a case of 29 year-old male patient who presented with genitourinary myiasis caused by Psychoda albipennis (Diptera: Nematocera), was presented. The patient was admitted to the hospital with the complaints of urinary incontinence of one week duration and presence of small, thin, motile, grayishwhite objects in his urine. Physical examination, blood and urine examination and stool microscopy revealed no pathology. No growth was detected in his urine culture. The examination of discharged larva in urine sample at Refik Saydam National Public Health Agency, Parasitology Laboratory led to the diagnosis of urogenital myiasis. No risk factor was identified in the patient who had proper hygienic conditions, was living in urban area and was of high socioeconomic status. This case was presented to withdraw attention to myiasis which is frequent in Turkey, however, is usually overlooked.

Gingival Myiasis on Oral Squamous Cell Carcinoma in Turkey: A Case Report

Myiasis is a disease caused by tissue invasion of diptera larvae and eggs. Oral myiasis is mostly related to old age, poor oral hygiene, suppurative lesions, anatomical disorders and cancer cases. Oral squamous cell carcinoma (OSCC) is an important risk factor for myiasis. This report presents the case of an 82-year-old woman who presented with gingival myiasis developing on the background of OSSC. The patient was diagnosed with OSSC in the hospital. Myiasis larvae were identified and sent to the National Parasitology Reference Laboratory for identification. Thus, development of myiasis on OSCC background was shown in Turkey for the first time. Myiasis larvae have been identified as the 3rd phase of the larvae Sarcophaga sp. development (Diptera: Sarcophagidae). As a result, myiasis cases are sporadic in Turkey, and it can be avoided by controlling fly population and by paying attention to hygiene. Controlling myiasis is an important public health problem and should be considered in a single health concept, as it causes health problems in both humans and animals. The findings of this case will draw attention to the importance of dealing with myiasis factors, which is a public health problem.

Seroprevalence of Encephalitozoon cuniculi, Francisella tularensis and Toxoplasma gondii in Zoonotic Diseases in European Hares (Lepus europaesus)

Objective: European hares (Lepus europaeus) are among the most important animals that are connected with humans in many countries and natural life. Hares are important for public health, since they carry many zoonotic diseases, such as Encephalitozoon cuniculi, Francisella tularensis and Toxoplasma gondii. The study aimed to determine the seroprevalence of Encephalitozoon cuniculi, Francisella tularensis and T. gondii and the potential zoonotic risk posed by hares that live in provinces of Turkey. Methods: Blood samples were collected from hares during the official hunting season. Serum samples were examined serologically by enzyme-linked immunosorbent assay for E. cuniculi, Sabin-Feldman dye test was used to examine T. gondii, while micro-agglutination test was used to examine F. tularensis. Results: Of the total of 42 hares examined, one (2.4%) was found positive for E. cuniculi, two (4.8%) were found positive for T. gondii and one (2.4%) was found positive for F. tularensis. Conclusion: Anti-T. gondii and anti-E. cuniculi antibodies were serologically detected in hares for the first time in Turkey. Furthermore, this is the first study reporting the seropositivity of F. tularensis infection in hares.

Evaluation of Seropositivity of Toxoplasmosis Suspected Patients Admitted to the National Parasitology Reference Laboratory Between 2009 and 2019

Objective: Toxoplasmosis caused by Toxoplasma gondii (T. gondii), which is an obligatory intracellular parasite, is a worldwide zoonotic parasitic disease. In this study, results of T. gondii test conducted between January 2009 and May 2019 were analysed. This study aimed to evaluate the results of T. gondii test of patients who were admitted to the General Directorate of Public Health, National Parasitology Reference Laboratory between 2009 and 2019. Methods: The results of anti-T. gondii IgG, IgM, IgG avidity and Sabin-Feldman dye tests (SFDT), which are used to detect the presence of T. gondii, were examined. ELISA was used for anti-T. gondii IgG, IgM and IgG avidity tests. SFDT, which is the reference test in the diagnosis of T. gondii, is still the gold standard. In addition to laboratory analyses, information on gender, age, city of origin, year distribution of all cases and type of sample sent was also collected. Results: Of the 2.778 patients evaluated, 25.4% were males and 74.6% were females. Moreover, 47.1% and 10.2% of the patients were positive for anti-T. gondii IgG and anti-T. gondii IgM antibodies, respectively. In SFDT, 1.228 (52%) patients were found to be positive, including 319 (59.4%) of 537 men and 909 (49.8%) of 1.824 women. In this 10-year study, the most common seropositivity titre of SFDT was at the level of 1/64. In our study, IgG levels were found to be positive in all cases in which IgG avidity was studied when all the cases in which all three of the anti-T. gondii IgG, IgM and IgG avidity tests were studied together in one patient were evaluated. In addition, of the 293 patients with positive anti-T. gondii IgG, 62.8% had high avidity, 24.2% had a limit value and 13% had a low avidity. In cases involving both mother and baby, anti-T. gondii IgG and IgM seropositivity rates were 80% and 5% for both, respectively. These high rates support the transfer of antibodies from the mother to the baby. Regarding the distribution of provinces from which the samples originated, the highest number of cases came from Ankara (80.7%). Blood is the most predominant sample, followed by cerebrospinal fluid. Conclusion: T. gondii maintains its importance in public health, owing to its high positivity rates. This study, in which 10-year data were collected, showed that despite an increase in awareness, high seropositivity still continues. Therefore, systematic collection and evaluation of laboratory analysis results for toxoplasmosis diagnosis will contribute in taking control measures.

Whole-genome sequencing of a Toxoplasma gondii strain from a Turkish isolate using next-generation sequencing technology.

PURPOSE: Toxoplasma gondii is an intracellular parasite that can affect all vertebrae and is the causative agent of toxoplasmosis. At present, the United States CDC (Centers for Disease Control and Prevention) recognizes this infection as a neglected disease. Toxoplasma gondii infection profiles exhibit differences because of the different regional and climatic responses to these parasites in Turkey, and these protozoan infections are notably common in this country. In this study, we attempted to obtain the whole-genome sequence of T. gondii using next-generation sequencing technology. METHODS: Toxoplasma gondii isolates were isolated from an infant with congenital toxoplasmosis by Ekmen et al. (1974) in Ankara, Turkey. Whole-genome sequencing (WGS) was performed using the Illumina HiSeq 2500 and HiSeq SBS Kit v2. A T. gondii library was created on this device in the initial stage. After the completion of the library phase, sequence analysis was begun with a next-generation sequencing device. The resulting fragments were combined using paired-end (PE) reading and converted into a single DNA fragment. Bioinformatic analysis was performed using the Geneious 2.1. RESULTS: In our study, WGS was successfully performed on T. gondii. The T. gondii whole-genome sequence has a coverage value of 50x, a size of 61,5763 Mb and a GC ratio of 52.6%. Data from this sequence were submitted to the National Center for Biotechnology Information (NCBI) GenBank (www.ncbi.nlm.nih.gov) database under the name Toxoplasma gondii TR01 (TG_TR01). The accession number of the genome obtained in this study is WOEV00000000.1. The biological sample access number is SAMN13338796. The genome of the T. gondii strain obtained in this study was compared with the reference genome, and 8312 CDSs (coding sequences), 183 tRNAs, 294 rRNAs and 8789 genes were identified. Among the 8312 CDSs, 4284 encoded hypothetical proteins (hypothetical protein CDSs/proteins of unknown function). The entire genome sequence of T gondii TR01 was compared with that of Toxoplasma gondii ME49. The results of this comparison demonstrate that the analyzed genome was 99,98% similar to the reference genome. The accession numbers of 14 chromosomes belonging to the genome sequences of T. gondii TR01 (TG_TR01) are CM019722.1, CM019723.1, CM019724.1, CM019725.1, CM019726.1, CM019727.1, CM019728.1, CM019729.1, CM019730.1, CM019731.1, CM019732.1, CM019733.1, CM019734.1, and CM019735.1. CONCLUSION: In this study, a whole-genome sequences of T. gondii was conducted for the first time in Turkey. The analyzed strain was named T. gondii TR01. The data obtained from this study may contribute to a better understanding of T. gondii. T. gondii is an important pathogen with an unusual population structure. Although T. gondii is highly zoonotic and has a complicated life cycle, some strains of this parasite have exhibited high genetic sequence similarity, and our study supports this knowlegde. The characterization of this strain may be very useful for the scientific community of our country and may help to establish a foundation for further research investigating the genome of T. gondii.

Is there a relationship between Toxoplasma gondii infection and idiopathic Parkinson's disease?

Idiopathic Parkinson's disease defines a group of Parkinson's disease (PD) of which the aetiology is unknown but an underlying brain disease is suspected. We selected patients of this subgroup of PD and investigated the seropositivity rate for anti-Toxoplasma IgG antibody by Sabin-Feldman dye test (SFDT). By measuring seropositivity in PD patients, we searched for a probable relationship between Toxoplasma gondii infection and idiopathic PD incidence. Fifty patients diagnosed with idiopathic PD and 50 healthy volunteers were included in the study. Blood samples were taken from all 100 participants and anti-T. gondii antibody titres were investigated using SFDT. Anti-T. gondii antibodies were detected at a titre of >or=1/16 in 25 of the 50 patients (50%) and in 20 of the control group (40%). No higher antibody titre was found in the control group. In conclusion, despite the emerging literature on a possible relationship between T. gondii infection and neurological disease, and the high anti-T. gondii seropositivity found in our PD patients, we did not detect any statistically significant association between T. gondii and idiopathic PD.

Detection and genetic diversity of Bartonella vinsonii subsp. berkhoffii strains isolated from dogs in Ankara, Turkey.

Bartonella vinsonii subsp. berkhoffii has been identified as an important pathogen in dogs and is an emerging pathogen in people with zoonotic potential. This study aimed to isolate Bartonella spp. in 250 blood samples collected from dogs in the province of Ankara, Turkey, between October 2006 and March 2007. The typing of the 23 isolates was carried out by PCR for citrate synthase (gltA) and the 16S-23S intergenic transcribed spacer (ITS). The prevalence of bacteremia was 9.2% in 250 samples. Among 170 shelter dogs, 21 (12.4%) were bacteremic for B. vinsonii subsp. berkhoffii, while the B. vinsonii subsp. berkhoffii bacteremia rate was 5% (2/40) for the stray dogs. B. vinsonii subsp. berkhoffii was not isolated from the pet dogs. The prevalence of bacteremia was found to be 25.5% (13/51) in shelter dogs aged less than one year old. All of the isolates were identified as B.vinsonii subsp. berkhoffii genotype III. In some isolates, MseI digest for gltA was found to be different from the American and European strains due to a single nucleotide change.

The role of latent toxoplasmosis in the aetiopathogenesis of schizophrenia--the risk factor or an indication of a contact with cat?

We assessed IgG antibody to Toxoplasma gondii in 300 inpatients with schizophrenia (SG), 150 outpatients with anxiety and depressive disorders (PCG), and 150 healthy blood donors (HCG). Seropositivity rates were 60.7% for SG, 36.7% for PCG, and 45.3% for HCG (p<0.001). The seropositivity rate for anti-Toxoplasma IgG antibodies in SG was significantly higher that in PCG (chi2 = 23.11, OR = 2.66, p = 0.001) and HCG (chi2 = 9.52, OR = 1.86, p = 0.002). Among SG, 85% of those who reported close cat contact had IgG antibodies to T. gondii. Close cat contacts were reported by 59% of SG, 6% of PCG, and 9% of HCG (p<0.001). There was a nonsignificant positive association between toxoplasmosis and schizophrenia for people with a contact with a cat (OR = 2.221, p = 0.127, CI95 = 0.796-6.192), and significant negative association between toxoplasmosis and schizophrenia for people without contact with a cat (OR = 0.532, p = 0.009, CI95 = 0.332-0.854). Close cat contact (OR = 2.679, p<0.001), 51-65-year age group (OR = 1.703, p<0.001) and education [illiterate+primary (OR = 6.146, p<0.001) and high school (OR = 1.974, p = 0.023)] were detected as independent risk factors in multivariate logistic regression. The effect of toxoplasmosis on risk of schizophrenia disappeared in the complex model analyzed with multivariate logistic regression. In conclusion, our data suggest that the toxoplasmosis has no direct effect on the risk of schizophrenia in Turkey but is just an indication of previous contacts with a cat.

[Fever of unknown origin and detection of Bartonella henselae IgG seropositivity: a case report]. / Bartonella henselae gingivostomatitli bir kedinin sahibinde nedeni bilinmeyen ates öyküsü ve B.henselae IgG seropozitifliginin saptanmasi

Bartonella henselae, is a gram-negative bacterium which causes cat scratch disease (CSD) in man. There are sporadic case reports of CSD in Turkey. Cats play an important reservoir role for B.henselae transmission to man. In this report, a cat owner with fever of unknown origin was presented. Bartonella spp. was isolated from the blood culture of cat which had chronic progressive gingivostomatitis. B.henselae was identified by amplification of a region of citrate synthase (gltA) gene by using polymerase cha-in reaction and typed as genotype I by restriction fragment length polymorphism method. Following this identification the cat owner was investigated for the history of CSD and it was learned that he had a history of fever of unknown origin. The investigation of the patient's serum for the presence of specific B.henselae antibodies by immune fluorescence antibody test (Vircell, Spain) revealed B.henselae IgG type antibodies at a titer of 1:128. Gingivostomatitis in cats may act as a reservoir for Bartonella infection. Thus during the evaluation of patients with fever of unknown origin, Bartonella infections should be considered and possible contact with cats/dogs should be investigated.

Conjugation and Characterization of Latex Particles with Toxoplasma gondii-specific Immunoglobulin Y Antibodies for Diagnostic Aim and Evaluation Efficiency in In Vitro Culture.

Toxoplasma gondii is a parasite that causes severe health problems in the world. Toxoplasmosis, an infection caused by T. gondii, leads to high risk of mortality in patients with immunodeficiency, transplantation, and cancer. Besides that, it causes miscarriages in pregnancy, various abnormalities such as hydrocephalus in infants and congenital diseases. Because the clinical indication of the disease is not specific, it is confused with many diseases, and this leads to the necessity of directly detecting the presence of the toxoplasmosis. Therefore, various diagnostic assays are needed for the diagnosis of the disease. Amongs them, latex agglutination assay is widely used for the detection of specific antibodies or antigens in samples. Latex particles are coated with immunogenic molecules (antigens) to detect antibodies in the blood or used to identify antigens when coated with specific antibodies. In both, aggregation of latex particles results in agglutination. Monoclonal antibodies are often used in latex agglutination assay as in other diagnostic methods. However, monoclonal antibodies can be produced in low quantities at a high cost. Besides, to produce monoclonal antibodies, an experienced staff, a well-equipped cell culture laboratory, a long period of time, and a burdened budget are needed. In recent years, as an alternative to monoclonal antibodies, immunoglobulin Y (IgY) antibodies, which are obtained from chicken eggs, and specifically produced against desired antigenic constructs, have become quite attractive in terms of both low cost and abundant production without requiring infrastructure. In contrast, the latex assay based on IgY antibodies for use in the diagnosis of T. gondii has not been developed. This study aimed to conjugate T. gondii-specific IgY antibodies to latex particles, characterize the particles by Fourier transform infrared spectroscopy, scanning electron microscopy, and spectroscopic methods, and finally demonstrate the interaction with T.gondii parasites in culture with scanning electron microscopy analysis.

Evaluation of Malaria Blood Smears Sent to National Malaria Reference Laboratory

Objective: Although the disease has been eliminated in Turkey malaria continues to be a threat due to increase in the number of people coming from or going to countries where the disease is endemic. In this study, we aimed to evaluate blood smears sent to the National Malaria Reference Laboratory within the malaria surveillance system. Methods: From March 2016 to July 2018 a retrospective study was conducted to compare the results of Malaria Reference Laboratory and Public Health Laboratories. A total of 16.827 blood stains were sent to our laboratory for approval. Results: In Public Health Laboratories, 315 (1.88%) of the smears were positive, 16.510 (98.12%) were negative, and in the National Malaria Reference Laboratory 252 (1.50%) were positive, 16.466 were negative. In the Public Health Laboratories, one of the two samples considered to be malaria suspected was positive in the National Malaria Reference Laboratory and one was negative. In Public Health Laboratories 35.88% of smears were P. falciparum, 27.30% were Plasmodium spp., 20.96% were P. vivax, 14.92% were mixed infection, 0.63% were P. malariae, 0.31% were P. ovale, and in the Reference Laboratory 49.60% were Plasmodium spp., 29.37% were P. falciparum, 16.27% were P. vivax, 4.36% were mixed infection, 0.40% were P. malariae. Conclusion: In order to malaria surveillance system to be maintained in a healthy manner, preparation, staining, coding, packaging, transportation of blood slides is very important. Also if necessary, continuing training of laboratory staff working in malaria diagnosis is crucial.