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.

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.

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.

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.

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.

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.

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.

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.

[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.

Molecular Survey of Babesia microti (Aconoidasida: Piroplasmida) in Wild Rodents in Turkey.

Babesia microti (Aconoidasida: Piroplasmida) (Franca, 1910) is an important tick-borne zoonotic parasite with rodents serving as reservoir hosts. In the present study, 536 rodents were captured from Burdur, Bartin, Giresun, and Yozgat provinces of Turkey between the years 2010 and 2012, and blood samples were examined for the presence of Babesia spp. using conventional PCR which targeted the 18S rRNA gene. The sequence analysis of PCR amplicons was tested for B. microti as well as for Hepatozoon spp., and Sarcocystis spp. Overall, 5.8% of the rodents were positive for B. microti: 41% in Myodes glareolus, 7.7% in Chionomys roberti, and 2% in Apodemus spp., whereas no Babesia DNA was detected in Mus macedonicus and Microtus spp. Six rodents were positive for Hepatozoon spp. and one rodent was positive for Sarcocystis spp. Overall, 14.9 and 4.5% of rodents captured from Bartin and Giresun provinces, respectively, were PCR positive for B. microti, whereas none of rodents captured in Burdur and Yozgat were positive for Babesia spp. The sequence data of B. microti from rodents revealed that all sequences belonged to the zoonotic genotype. Sequences of B. microti obtained from rodents of the Bartin province were genotypically closer to European isolates, whereas those obtained from rodents of the Giresun province were closer to Russian and Mongolian isolates.

Investigation of Anti-Toxoplasma gondii Antibodies in Cats Using Sabin-Feldman Dye Test in Ankara in 2016

Objective: Toxoplasmosis, in which obligate intracellular protozoa Toxoplasma gondii (T.gondii) is the causative organism, is a multisystemic disease that can be seen all over the world and can impair all vertebrates. The only hosts known for T.gondii are members of Felidae family. Our study aimed to determine anti-Toxoplasma gondii antibodies with Sabin-Feldman Dye Test (SFDT) in cats in Ankara. It's aimed to evaluate the current situation in terms of Toxoplasmosis spread by comparing our findings with previous studies in the same region. Methods: Rh strain of Toxoplasma used in our study is maintained in our laboratory. SFDT is still accepted as the gold standard. Material of the study was obtained by taking blood samples from cats who were admitted to the clinics between March 2016 and October 2016 in Ankara. Blood samples were inactivated and measurements were done with SFDT 1/4, 1/16, 1/64, 1/256, 1/1024 titers. Results: SFDT resulted positive in 56 (43.4%) cats at a dilution of 1/16, in 7 (5.4%) cats at a dilution of 1/64, in 23 (17.8%) cats at a dilution of 1/256 and negative in 43 (33.3%) cats. Comparison of demographic data with SFDT results showed that positive test results did not differ according to gender and age (P=0.803 and P=0.991, respectively). Seropositivity was higher in stray cats than house cats (P<0.001). Test results were negative in the cats that fed only by commercial dry food (P<0.001). Positivity in hunter cats was more than in non-hunters (P<0.001). Conclusion: Seropositivity was detected in 66.6% of the cats, which was quite a high rate. As a result, taking precautions in terms of Toxoplasma for stray cats that are hunting and feeding naturally is a necessity for public health.

Role of NETs in the difference in host susceptibility to Toxoplasma gondii between sheep and cattle.

The main aim of this study was to compare extracellular traps (NETs) formation by polymorphonuclear neutrophils (PMNs) of cattle and sheep when exposed to T. gondii tachyzoites in vitro. The effects of parasite concentrations and different incubation periods on NETs development in cattle and sheep PMNs were studied. The effect of NET structures on host cell invasion by tachyzoites was also studied. This is the first report of NETs development by sheep and cattle PMNs against T. gondii in vitro. T. gondii-induced extracellular DNA production from PMNs was dependent on tachyzoite concentrations and incubation time in both sheep and cattle. Many nuclear and cytoplasmic changes were observed in sheep and cattle PMNs after exposure to T. gondii tachyzoites. The typical appearance of NETs, with MPO, NE and histone (H3) attached to extracellular DNA, was observed. Tachyzoites were entrapped within this structure. Myeloperoxidase (MPO) activity was higher in the cattle PMN-tachyzoite co-cultures than sheep. NETs structures released from sheep PMNs caused mechanical immobilisation of T. gondii tachyzoites, however, NET structures released from cattle PMNs may be lethal to tachyzoites. Bovine MPO may have a lethal effect on T. gondii tachyzoites in vitro during a 3h incubation. Besides other mechanisms that effect on host susceptibility to T. gondii in sheep and cattle, extracellular traps formation as a part of immunological reactions may be play a role in host susceptibility to T. gondii.

Experimental Toxoplasmosis in Japanese Quails (Coturnix coturnix japonica).

OBJECTIVE: In this study, forming of experimental toxoplasmosis in quails; clinical, pathological, and serological determination of tissue lesions and bioassay techniques, which were aimed to compare them and determine pathogenesis. METHODS: A total of 120 one-year-old female quails (Coturnix coturnix japonica) were divided into oral infection, parenteral infection, and control groups. The oral group was infected with 0.5 ml inoculum suspension containing 106 tachyzoites of Toxoplasma gondii, whereas the control group was administered 0.5 ml of saline. The parenteral group was further divided into the following four subgroups: intraperitoneal, intramuscular, intravenous, and cloacal. The quails of the parenteral group were also divided into two groups and one by control group within itself for the 105 and 104 doses of the tachyzoite inoculums. RESULTS: Because of acute toxoplasmosis, death occurred in a quail that as intramuscularly infected with 105 tachyzoites; the quail exhibited neurological clinical symptoms such as torticollis, ataxia, and tremor. In histopathologic examination, T. gondii tissue cysts were detected in infected quails that were intramuscularly infected with 105 tachyzoites. Mouse trials were conducted using tissues of seropositive quails and isolated from peritoneal fluids infected mice. By Sabin-Feldman dye test and indirect hemagglutination test, seropositivity was observed in quails infected with 105 and 104 tachyzoites. CONCLUSION: Similar studies and subclinical cases, which may overlooked was concluded for diagnosis of toxoplasmosis with useful bioassay applications and serological tests.

[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.

In Vitro Efficacy of the Ankaferd Galenic Hemostatic Extract as a Germicidal Agent.

BACKGROUND: Hydatid cysts are encountered frequently in regions endemic with livestock. The basic treatment for a hydatid cyst is total surgical removal of the cyst and its inner contents. Hypertonic NaCl or diluted betadine solution are used as germicidal agents for most hydatid surgeries. However, the germicidal efficacy of the Ankaferd Blood Stopper® (ABS) has not been investigated. Thus, we compared the efficacy of ABS for hydatid cysts with that of other germicidal agents. METHODS: Lung and liver tissues containing hydatid cyst liquid were collected from slaughterhouses. Six samples of each cyst were randomly allocated into different groups as follows: 20% hypertonic NaCl, betadine solution, ABS, 20% liquefied Andazole solution, 0.1% eosin, and distilled water. All groups were examined microscopically at 5, 10, and 15 min after treatment began to determine protoscolece viability rates. RESULTS: The most efficacious germicidal agent at 5 min was ABS, and betadine and hypertonic NaCl had similar efficacies. Betadine, ABS, and hypertonic NaCl showed similar efficacies at 15 min. CONCLUSION: ABS was an effective germicidal agent to treat hydatid cysts.

[Evaluation of Cystic Echinococcosis Suspected Patients Applied to National Parasitology Reference Laboratory of Public Health Institution of Turkey Between 2009-2013]. / Gaziantep Üniversitesi Tip Fakültesi Leishmaniasis Tani ve Tedavi Merkezine Basvuran Kutanöz Leishmaniasis Olgularinin Degerlendirilmesi.

OBJECTIVE: Cystic echinococcosis (CE) caused by the metacestode form of Echinococcus granulosus is an important public health problem common in our country. In this study, anti-E. granulosus antibodies were aimed to investigate in the serum samples of CE suspected patients who applied to the National Parasitology Reference Laboratory of Public Health Institution of Turkey. METHODS: In the study, serum samples of 2921 patients which were sent to our laboratories from different hospitals between 1 January 2009 and 31 December 2013 were evaluated with at least one of the following tests: Enzyme-Linked Immunosorbent Assay (ELISA), Indirect Hemaglutination Assay (IHA) and Western Blot (WB) techniques. RESULTS: Four hundred thirty nine (15.03%) of inspected 2921 samples were determined seropositive with at least one of the methods. When the results analyzed by gender, 13% of males and 16.40% of females were found positive. Examined the distribution of the results by years, with a maximum of 25% positivity was observed in 2009. Compatibility was determined at the rate of 91.4% among ELISA and IHA results; also 89.7% among WB and the other tests results. CONCLUSION: Despite the gradual decreases the CE in Ankara and its surroundings, it is still continues to be a major public health problem. Essential prevention and control measures should be taken to reduce the prevalence of the disease. Also in the diagnosis of the disease, more reliable results can obtained with applying two tests (ELISA/IHA) together and confirm the positivity with WB.

Capillaria Hhepatica in mouse (Apodemus flavicollis) from Giresun Province of Turkey.

Capillaria hepatica is a nematode with worldwide distribution, which can cause parasitic hepatitis both in animals and humans. A mouse (Apodemus flavicollis), trapped in Giresun Province was diagnosed as having capillariasis due to the characteristic eggs found in its liver. This is the first reported case of mouse capillariasis in this part of the country. Due to the fact that capillariasis is a zoonotic disease, humans might be also infested; therefore, further investigations are needed.