Molecular Prevalence of Microsporidia Infection in Patients with Lung Cancer.

Infections are still among the most important causes of morbidity and mortality in patients with lung cancer, which has the highest rate of cancer-related deaths in the world. Microsporidia, which are opportunistic parasitic fungi, primarily localize to the intestine by ingestion but can disseminate to the respiratory tract or can be acquired by spore inhalation. Cancer patients are at higher risk for microsporidia, a life-threatening infection, than the normal population is. We aimed to characterize the prevalence of microsporidia infection for the first time by evaluating the intestinal and respiratory tracts of patients with lung cancer. In this study, we investigated 98 patients with lung cancer and 103 healthy individuals for microsporidia infection and evaluated the clinical findings of patients who were found to be positive. Sputum and stool samples were tested by microscopic examination, in addition to pan-microsporidia and genus-specific polymerase chain reactions. Nine patients with lung cancer had positive results for microsporidia (9.2%), which was significantly higher than the rate in healthy individuals (P = 0.008), and most of them had clinical findings. Among these positive patients, polymerase chain reaction revealed microsporidia in the sputum samples of seven patients, the stool sample of one patient, and both the sputum and stool samples of one patient. Encephalitozoon cuniculi was identified as the predominant pathogen in 87.5% (7/8) of positive sputum samples. Microsporidia infection was significantly associated with advanced stages of cancer. However, in the control group, Encephalitozoon intestinalis was detected in the stool sample of an individual without clinical symptoms. Microsporidia, especially E. cuniculi, should be considered as a cause of respiratory tract infection as well as intestinal infection in cancer patients and should be screened in respiratory samples of these patients when they have pulmonary symptoms.

Prevalence of head lice in two socio-economically different schools in the center of Izmir City, Turkey.

OBJECTIVE: The well-known and common infestation caused by Pediculus humanus capitis is an important public health and a social issue in many communities in the world. The aim of this study was to compare the head louse infestation rate in two schools having pupils from different socio-economic levels in the city center of Izmir. METHODS: The pupils aged between 6 and 11 years, were screened for the presence of eggs and nymph/adult lice using a fine-tooth head louse comb. RESULTS: A total of 88 and 126 pupils from the schools with low and medium socio-economic level were screened and 24 (27.2%) and 5 (3.96%) of them were found to be positive for head lice, respectively. Overall, the infestation rate among girls was 3.14 times higher than in boys. CONCLUSION: Head louse infestation is a significant public health problem among primary schools. Increasing the knowledge about pediculosis and self-hygiene would be helpful in successfully reducing head louse infestation in the school setting. School authorities must encourage the parents to look for head lice routinely and a "school nurse" system is needed for effective head louse control in the schools.

Genetic diversity and structure in Leishmania infantum populations from southeastern Europe revealed by microsatellite analysis.

BACKGROUND: The dynamic re-emergence of visceral leishmaniasis (VL) in south Europe and the northward shift to Leishmania-free European countries are well-documented. However, the epidemiology of VL due to Leishmania infantum in southeastern (SE) Europe and the Balkans is inadequately examined. Herein, we aim to re-evaluate and compare the population structure of L. infantum in SE and southwestern (SW) Europe. METHODS: Leishmania strains collected from humans and canines in Turkey, Cyprus, Bulgaria, Greece, Albania and Croatia, were characterized by the K26-PCR assay and multilocus enzyme electrophoresis (MLEE). Genetic diversity was assessed by multilocus microsatellite typing (MLMT) and MLM Types were analyzed by model- and distance- based algorithms to infer the population structure of 128 L. infantum strains. RESULTS: L. infantum MON-1 was found predominant in SE Europe, whilst 16.8% of strains were MON-98. Distinct genetic populations revealed clear differentiation between SE and SW European strains. Interestingly, Cypriot canine isolates were genetically isolated and formed a monophyletic group, suggesting the constitution of a clonal MON-1 population circulating among dogs. In contrast, two highly heterogeneous populations enclosed all MON-1 and MON-98 strains from the other SE European countries. Structure sub-clustering, phylogenetic and Splitstree analysis also revealed two distinct Croatian subpopulations. A mosaic of evolutionary effects resulted in consecutive sub-structuring, which indicated substantial differentiation and gene flow among strains of both zymodemes. CONCLUSIONS: This is the first population genetic study of L. infantum in SE Europe and the Balkans. Our findings demonstrate the differentiation between SE and SW European strains; revealing the partition of Croatian strains between these populations and the genetic isolation of Cypriot strains. This mirrors the geographic position of Croatia located in central Europe and the natural isolation of the island of Cyprus. We have analysed the largest number of MON-98 strains so far. Our results indicate extensive gene flow, recombination and no differentiation between MON-1 and MON-98 zymodemes. No correlation either to host specificity or place and year of strain isolation was identified. Our findings may be associated with intensive host migration and common eco-epidemiological characteristics in these countries and give valuable insight into the dynamics of VL.

[An epidemiological study on canine leishmaniasis (CanL) and sand flies in Northern Cyprus]. / Kuzey Kibris'ta Kanin Leishmaniasis ve Kum Sineklerinin Epidemiyolojisi.

OBJECTIVE: In this study, the investigation on the prevalence of canine leishmaniasis (CanL) and sand fly species incriminated as potential vectors of leishmaniasis in the northern part of the Cyprus were aimed. METHODS: This research was conducted in two periods; 2004 and 2012. Serological (IFAT and rK39) and molecular (PCR) tests were performed on 83 dog blood samples during the 2004 survey. PCR was performed using primers 13A/13B targeting kinetoplastid minicircle constant region. Genomic Nested-PCR was applied using R221/R332 and R323/333 primers for 5 clinically suspected dog samples in 2012. Sand flies were collected from the Lapithos town and Kyreniae province using CDC light traps and midgut dissection was done for the presence of Leishmania parasites during the 2012 survey. RESULTS: Three (3.61%) out of 83 dogs were found to be positive for CanL in 2004, while 3 out of 5 clinically suspected dogs were positive in 2012. In total 296 female sand flies were dissected and 9 species belonging to Phlebotomus and Sergentomyia genera were determined. No promastigote was found in the dissected females. CONCLUSION: The results obtained in two different periods showed that the importance and risk of canine disease are increasing in the northern Cyprus and further studies should be performed in northern Cyprus for determining the incidence of canine and human leishmaniasis.

A real-time ITS1-PCR based method in the diagnosis and species identification of Leishmania parasite from human and dog clinical samples in Turkey.

Human visceral leishmaniasis (VL) caused by L. infantum and cutaneous leishmaniasis (CL) caused by L. tropica and L. infantum have been reported in Turkey. L. infantum is also responsible for canine leishmaniasis (CanL) and it is widely common in the country. The main aim of the present study was to design a real-time PCR method based on the internal transcribed spacer 1 (ITS1) region in the diagnosis of all clinical forms of leishmaniasis in Mediterranean, and to identify the species directly from clinical samples. Totally, 315 clinical specimens, human/canine visceral (blood, bone marrow, lymph node) and cutaneous (lesion aspiration) samples, and 51 Turkish Leishmania isolates typed by isoenzymatic method were included in the study. For optimization, DNA samples of the 34 strains were amplified by conventional ITS1-PCR and then sequenced for designing the primers and probes, allowing the species identification. Following the validation with the isolates, the test was applied on clinical samples and melting temperatures were used for genotyping. A group of PCR products were further sequenced for confirmation and assigning the inter- and intraspecies heterogeneity. The diagnosis of leishmaniasis is successfully achieved by the new real-time PCR method, and the test identified 80.43% of human and canine VL samples as L.infantum and 6.52% as L.tropica; 52.46% of CL samples as L. infantum and 26.90% as L. tropica. In 13.04% of visceral and 20.62% of cutaneous samples, two peaks were observed. However, the higher peak was found to be concordant with the sequencing results in 96.96%, in terms of species identification. The real-time ITS1 PCR assay clearly identified the leishmanial species in 81.58% of all clinical samples. Genotypic variations of Leishmania parasites in Turkey within species and intraspecies were observed, and L. tropica is also found as causative agent of human and canine VL in Turkey.

[Detection of entomopathogen nematode [EPN - sand flies (Phlebotomus tobbi)] caught in the wild in Aydin, Kusadasi town and its assessment as a biological control agent]. / Kusadasi, Aydin'da Dogadan Yakalanan Kum Sineklerinde (Phlebotomus tobbi) Saptanan Entomopatojen Nematod [EPN] ve Biyolojik Kontrol Ajani Olarak Kullaniminin Degerlendirilmesi.

OBJECTIVE: In this study, the midgut of the sand flies investigated with direct method for the presence of parasites and other organisms. METHODS: Wild sand flies collected in Kusadasi Town-Aydin, were dissected and midgut contents were examined by light microscopy. After midgut dissection, the head and genitalia of sand fly specimens were clarified and mounted for species identification. RESULT: During the study, a total of 1027 sand flies were dissected. Eight and two species belonging to Phlebotomus and Sergentomyia genera were determined, respectively. Phlebotomus tobbi was found to be most abundant species (61.34%). A third stage of infective Entomopathogen Nematode belonging to Steinernematidae family was observed in the hemocoel of one specimen of P. tobbi during the dissection process. CONCLUSION: This is the first finding related to entomopathogen nematodes found in sand flies in Turkey. In the study, the sand fly fauna was determined in Kusadasi Town. For the control of sand flies, entomopathogenic nematodes which are not harmful for non-target organisms, can be used instead of chemical insecticides that can cause unknown damage in the environment.

[Co-occurence of visceral leishmaniasis and lymphoma]. / Leishmaniasis ve lenfoma birlikteligi.

Clinicians have usually considered malignancies during follow up of patients who have infectious diseases as a pre-diagnosis. However, malignancy and an infectious disease are seen together more rarely, with the exception of immunosuppressed patients. This presentation is a case report followed up for fever of unknown origin. The patient was admitted to the hospital with the symptoms of fever, weight loss, abdominal pain and weakness. Anemia and hypergamaglobulinemia by biochemical analyses and splenomegaly by total body computed tomography were detected. Amastigotes were seen in bone marrow aspiration smears and promastigotes were isolated in NNN medium. At the end of the Liposomal Amphotericin B treatment, control bone marrow aspiration was applied. Leishmania amastigotes were not seen, while patient was diagnosed as diffuse B cell lymphoma pathologically.

Multilocus microsatellite typing (MLMT) of strains from Turkey and Cyprus reveals a novel monophyletic L. donovani sensu lato group.

BACKGROUND: New foci of human CL caused by strains of the Leishmania donovani (L. donovani) complex have been recently described in Cyprus and the Çukurova region in Turkey (L. infantum) situated 150 km north of Cyprus. Cypriot strains were typed by Multilocus Enzyme Electrophoresis (MLEE) using the Montpellier (MON) system as L. donovani zymodeme MON-37. However, multilocus microsatellite typing (MLMT) has shown that this zymodeme is paraphyletic; composed of distantly related genetic subgroups of different geographical origin. Consequently the origin of the Cypriot strains remained enigmatic. METHODOLOGY/PRINCIPAL FINDINGS: The Cypriot strains were compared with a set of Turkish isolates obtained from a CL patient and sand fly vectors in south-east Turkey (Çukurova region; CUK strains) and from a VL patient in the south-west (Kusadasi; EP59 strain). These Turkish strains were initially analyzed using the K26-PCR assay that discriminates MON-1 strains by their amplicon size. In line with previous DNA-based data, the strains were inferred to the L. donovani complex and characterized as non MON-1. For these strains MLEE typing revealed two novel zymodemes; L. donovani MON-309 (CUK strains) and MON-308 (EP59). A population genetic analysis of the Turkish isolates was performed using 14 hyper-variable microsatellite loci. The genotypic profiles of 68 previously analyzed L. donovani complex strains from major endemic regions were included for comparison. Population structures were inferred by combination of bayesian model-based and distance-based approaches. MLMT placed the Turkish and Cypriot strains in a subclade of a newly discovered, genetically distinct L. infantum monophyletic group, suggesting that the Cypriot strains may originate from Turkey. CONCLUSION: The discovery of a genetically distinct L. infantum monophyletic group in the south-eastern Mediterranean stresses the importance of species genetic characterization towards better understanding, monitoring and controlling the spread of leishmaniasis in this region.

Serological monitoring of paediatric visceral leishmaniasis By IFA and ELISA methods.

OBJECTIVE: Visceral leishmaniasis (VL) is endemic in all Mediterranean countries including Turkey, and children are at greater risk than adults in endemic areas. In VL patients, serological assays are considered to be sensitive for the diagnosis and/or follow up. The aim of this study was to assess the usefulness of serology for following up of childhood VL in Turkey. METHODS: Sera obtained from twenty parasitologically confirmed children with VL were tested using IFAT and ELISA. The patients were monitored clinically and serologically (range: 20-500 days) during and after treatment. All VL patients were treated with meglumine antimonate. RESULTS: Anti-Leishmania antibodies in successfully treated VL patients showed a steep decline but, in three patients who had relapsed, an increase was detected. Significantly lower values were observed after treatment with both serological techniques. Mean ELISA optical density values before and after treatment were: 0.78 ± 0.36 (0.26-1.76) and 0.38 ± 0.24 (0.09-0.83) respectively, (p < 0.001) and mean IFAT values (log10 transformed titers) before and after treatment were: 3.02 ± 0.90 (1.81-4.51) and 2.16 ± 0.75 (1.20-3.90) respectively, (p < 0.001). CONCLUSION: ELISA and IFAT are valuable not only for diagnosis but also for monitoring of drug therapy in childhood visceral leishmaniasis as rapid and non-invasive techniques.

[Comparison of polymerase chain reaction with wet mount and culture methods for the diagnosis of trichomoniasis]. / Trichomoniasis Tanisinda Polimeraz Zincir Reaksiyonu ile Mikroskobi ve Kültür Yöntemlerinin Karsilastirilmasi

OBJECTIVE: Wet mount, culture and staining methods are generally used in the diagnosis of trichomoniasis. Polymerase chain reaction (PCR) methods with different primer pairs have been tested and used for research in recent years. METHODS: In this study, T. vaginalis was tested for in the vaginal samples of 102 patients referred to obstetrics and gynecology polyclinic of Aydin Obstetrics and Children Hospital for various reasons, with direct microscopy, culture and PCR with primers targeting Tv-E650. In addition, PCR was applied to 20 T. vaginalis strains that were isolated from patients who were previously diagnosed with vaginitis. RESULTS: Of 102 samples, T. vaginalis was found to be positive in 2.94, 4.90 and 4.90% with wet mount, TYM medium and PCR respectively. The positivity rate reached 5.88% using the 3 methods together. All 20 strains isolated from patients with vaginitis were reported as positive by the PCR method. CONCLUSION: The wet mount had 60% sensitivity and 100% specificity, while PCR showed 80% sensitivity and 97.95% specificity when compared with the culture method, which is accepted as the "gold standard". The PCR method was performed for the first time as a diagnostic assay for trichomoniasis in this study and it is concluded that it can be used routinely for its diagnosis.

Spatial distribution of phlebotomine sand flies in the Aydin Mountains and surroundings: the main focus of cutaneous leishmaniasis in western Turkey.

An entomological survey was conducted to determine the spatial distribution of phlebotomine fauna and understand the effect of environmental factors. The entomological survey was carried out during 2006-2007 in a study area in the rural area of Aydin province, near the Kusadasi town where VL, CL, and canine leishmaniasis (CanL) are endemic. In 2006 and 2007, 132 locations were sampled using sticky traps mainly on embankments. Detailed environmental and meteorological information was also collected for each location. The results of entomological studies indicated that the probable vectors are Phlebotomus tobbi and P. neglectus for VL and CanL, and P. similis for CL in this western leishmaniasis focus. The data revealed a correlation between their presence and spatial variables such as altitude, sampling site location, and humidity. The distribution areas of probable vector species in this study area allowed the identification of risk levels, which may provide useful information to guide the leishmaniasis research in endemic regions.

Quantiferon-Leishmania as an epidemiological tool for evaluating the exposure to Leishmania infection.

The aim of the present preliminary study was to investigate the potential of measurement of IFN-γ secretion by T cells into blood plasma using QuantiFERON assay with leishmanial antigens to determine the presence of Leishmania infection. Blood samples from cured visceral (N = 18), and cutaneous (N = 20) leishmaniasis cases, and 20 healthy controls were tested. The IFN-γ responses to Leishmania major H2B and Leishmania infantum H2B antigens were detected from the majority of treated old visceral leishmaniasis cases, but not from controls. Future studies using larger groups will be required to establish the true potential of the assay for epidemiological screening of leishmaniasis.

[The investigation of Entamoeba gingivalis and Trichomonas tenax in a group of patients with periodontal disease]. / Periodontal Hastaligi Bulunan Kisilerde Dis Eti Plaklarinda Entamoeba gingivalis ve Trichomonas tenax Arastirilmasi.

The oral cavity is suitable for invasion of many microorganisms. Entamoeba gingivalis (E.gingivalis) and Trichomonas tenax (T.tenax) settle in the oral cavity of patients with poor oral hygiene and gingival disease. In the present study, two slide specimens were prepared from the cole region of the teeth of 46 persons for investigation of the parasites. One of the slide specimens was dried in the air while the other one put into fixative and they were stained with trichrome and Giemsa stains. The two staining methods were used for 36 samples and only Giemsa, for 10 samples. E. gingivalis was positive in 7 (19.44%) out of 36 samples stained by the trichrome stain while T. tenax was positive in one (2.17%) out of 46 samples stained by Giemsa stain. Parasitic infections were found to be positive in seven (21.2%) specimen from 33 patients with periodontal disease and in one (7.69%) specimen from 13 healthy controls. Dental policlinics are generally far from parasitology laboratories and microscopical wet mount examination can not be performed. Therefore dentists can send the specimens and have the parasites diagnosed with Giemsa and trichrome staining methods as an alternative to wet mount examination.

Detection and identification of old world Leishmania by high resolution melt analysis.

BACKGROUND: Three major forms of human disease, cutaneous leishmaniasis, visceral leishmaniasis and mucocutaneous leishmaniasis, are caused by several leishmanial species whose geographic distribution frequently overlaps. These Leishmania species have diverse reservoir hosts, sand fly vectors and transmission patterns. In the Old World, the main parasite species responsible for leishmaniasis are Leishmania infantum, L. donovani, L. tropica, L. aethiopica and L. major. Accurate, rapid and sensitive diagnostic and identification procedures are crucial for the detection of infection and characterization of the causative leishmanial species, in order to provide accurate treatment, precise prognosis and appropriate public health control measures. METHODS/PRINCIPAL FINDINGS: High resolution melt analysis of a real time PCR product from the Internal Transcribed Spacer-1 rRNA region was used to identify and quantify Old World Leishmania in 300 samples from human patients, reservoir hosts and sand flies. Different characteristic high resolution melt analysis patterns were exhibited by L. major, L. tropica, L. aethiopica, and L. infantum. Genotyping by high resolution melt analysis was verified by DNA sequencing or restriction fragment length polymorphism. This new assay was able to detect as little as 2-4 ITS1 gene copies in a 5 microl DNA sample, i.e., less than a single parasite per reaction. CONCLUSIONS/SIGNIFICANCE: This new technique is useful for rapid diagnosis of leishmaniasis and simultaneous identification and quantification of the infecting Leishmania species. It can be used for diagnostic purposes directly from clinical samples, as well as epidemiological studies, reservoir host investigations and vector surveys.

Leishmaniasis in Turkey: molecular characterization of Leishmania from human and canine clinical samples.

Human leishmaniasis, both visceral and cutaneous, and canine leishmaniasis have been reported in Turkey for centuries. However, the advent of new diagnostic tools during the last 30 years has led to the recognition that leishmaniasis is an important public health problem throughout the country. In most disease foci both canine and human leishmaniases exist together and identification of parasite species causing these diseases is a pre-requisite for understanding disease epidemiology. A total of 109 samples obtained from human and canine leishmaniasis cases were examined using internal transcribed spacer 1 PCR followed by restriction fragment length polymorphism analysis. Our results indicate that two species, Leishmania tropica and Leishmania infantum, are primarily responsible for cutaneous and visceral leishmaniasis, respectively, in Turkey. However, a new focus of human cutaneous leishmaniasis caused by L. infantum in Hatay region is described. This finding further stresses the importance of Leishmania species molecular characterization in prescribing appropriate therapy and understanding the disease's transmission in different endemic foci.

[Investigating the seroprevalance of leishmaniasis in four dog shelters in Antalya and its districts]. / Antalya Ili ve Ilçelerindeki Dört Köpek Barinaginda Leishmaniasis Seroprevalansinin Arastirilmasi.

Human visceral leishmaniasis (HVL) is endemic along the Aegean and Mediterranean coasts, while it occurs sporadically in other regions of Turkey. A relationship between canine leishmaniasis (CanL) and HVL has been detected and dogs have been found to be the reservoir of Leishmania infantum. In this epidemiologic study, the seroprevalence of CanL was investigated in dog shelters of four districts (Kepez, Kemer, Alanya and Gazipasa) of Antalya, in which visceral leishmaniasis and cutaneous leishmaniasis were observed according to the official records of The Ministry of Health of Turkey. Blood specimens of 176 dogs were examined and seropositive, borderline, seropositive, and negative results were detected in 14 (7.95%), 24 (13.63%), and 138 (78.4%) dogs, respectively. Weight and alopecia, onychogryphosis, and skin ulcers around the nose were observed in only two (14.2 %) dogs of seropositive cases. In conclusion, our data indicate that there is a potential danger for humans because L. infantum carrying dogs were detected in four districts of Antalya in the present study and also the appropriate vector spp. for this parasite has been reported in a previous one. Therefore, it would be advisable to perform new studies in order to determine the seroprevalence of CanL in dogs and the population size of vector spp. of L. infantum in other districts of Antalya.

[Seroprevalence of visceral leishmaniasis in stray dogs in Kocaeli]. / Kocaeli Sokak Köpeklerinde Visseral Leishmaniasis Seroprevalansi.

Dogs are the most important reservoir of visceral leishmaniasis (VL). A male child who lives in Köseköy in Kocaeli was diagnosed with VL. Since this child had never been outside Kocaeli, serum samples of 65 stray dogs were analyzed for canine VL using the indirect fluorescence antibody test (IFAT) and ELISA. Two dogs (3.07%) tested positive with both ELISA and IFAT. Leishmania amastigotes were observed in the lymph aspiration material from one of them. Growth was observed in NNN medium inoculated with lymph aspiration material from the other dog. This was the first study investigating canine VL prevalence in our city and plans were made for control of the disease.

Drug regimens for visceral leishmaniasis in Mediterranean countries.

Until the early 1990s, pentavalent antimony was the only documented first-line drug employed for the treatment of zoonotic visceral leishmaniasis (VL) in the Mediterranean, with reported cure rates exceeding 95% in immunocompetent patients. The emergence of antimony resistance in other endemic settings and the increase in drug options have stimulated re-evaluation of the current therapeutic approaches and outcomes in Mediterranean countries. A scientific consortium ('LeishMed' network) collected updated information from collaborating clinical health centres of 11 endemic countries of Southern Europe, Northern Africa and the Middle East. In contrast with the previous situation, VL is now treated differently in the region, basically through three approaches: (1) In Northern Africa and in part of the Middle East, pentavalent antimony is still the mainstay for therapy, with no alternative drug options for treating relapses; (2) In some European countries and Israel, both pentavalent antimony and lipid-associated amphotericin B (AmB) formulations are used as first-line drugs, although in different patients' categories; (3) In other countries of Europe, mainly liposomal AmB is employed. Importantly, cure rates exhibited by different drugs, including antimonials in areas where they are still in routine use, are similarly high (>/=95%) in immunocompetent patients. Our findings show that antimony resistance is not an emerging problem in the Mediterranean. A country's wealth affects the treatment choice, which represents a balance between drug efficacy, toxicity and cost, and costs associated with patient's care.

[Investigation of anti-Leishmania seroprevalence by different serologic assays in children inhabiting in the northwestern part of Turkey]. / Ulkemizin kuzeybatisindaki il ve ilçelerde yasayan cocuklarda anti-Leishmania seroprevalansinin farkli serolojik yöntemlerle arastirilmasi.

Human visceral leishmaniasis (VL) caused by Leishmania infantum in Mediterranean region is still an important public health problem in those countries including Turkey. The asymptomatic presentation of the infection in most of the cases leads to difficulties in the diagnosis and prevention of the spread of infection. The aim of this study was to investigate the seroprevalence of anti-Leishmania antibodies in children inhabiting in the northwestern part of Turkey. A total of 572 healthy children (260 girls, 312 boys) aged between 1-17 years old (mean age: 8.1 years) inhabiting in Eskisehir, Bilecik, Kutahya and Afyon provinces and their counties were included to the study with the informed consent obtained from their families. All serum samples were screened by a commercial ELISA (Leishmania Ab Panel, Cypress Diagnostics, Belgium), two in-house ELISA (with whole antigens and rK39 antigen) tests and an in-house indirect fluorescent antibody test (IFAT). The sera yielding a positive result by commercial ELISA, have been re-evaluated with rK39 dipstick test. Of 572 children, 5.2% (n = 30) were found positive with commercial ELISA, 3.8% (n = 22) with IFAT, 4.7% (n = 27) with in-house whole ELISA, and 3.6% (n = 21) with in-house rK39 ELISA. Of 30 commercial ELISA positive sera, 19 (63.3%) gave positive result also by dipstick test. This difference was attributed to the use of multiple antigens belonging to different Leishmania species in commercial ELISA test. The number of children who were seropositive with all of the tests were 15 (2.6%) and with at least three of the tests were 24 (4.2%). Most (76.7%) of the 30 seropositive children with commercial ELISA were found to be inhabiting in Iscehisar town of Afyon province (n = 16) and Sogut town of Bilecik province (n = 7) where canine VL was also endemic. Our results also indicated that 93.3% (28/30) of seropositive children were living in the rural parts of the study region. Thirty seropositive children were followed-up for one year, but none of them have exhibited clinical findings of VL. As a result, the rate of anti-Leishmania seropositivity (5.2% with at least one test, and 4.2% with at least three tests) in healthy children in the study area should not be ignored, and prevention strategies should be undertaken since our previous study have also showed that the rate of VL in dogs was high and the vector Phlebotomus species were determined in these areas of Turkey.

In vitro and in vivo activities of Haplophyllum myrtifolium against Leishmania tropica.

This study aimed to evaluate the in vitro and in vivo leishmanicidal activity of an endemic Turkish plant and compare its efficacy with a reference drug. In addition to the in vitro activities of the ethanol, acidified and alkaloid extracts and furoquinoline alkaloids skimmianine and gamma-fagarine, in vivo antileishmanial activitiy of the acidified extract of Haplophyllum myrtifolium Boiss. (Rutaceae) were investigated against Leishmania tropica (L. tropica), a causative agent of cutaneous leishmaniasis. All the extracts and pure compounds showed in vitro inhibitory activity against the promastigotes of. L. tropica. The in vitro 50% inhibitory concentrations of y-fagarine, acidified extract, ethanol extract, skimmianine and alkaloid extract against promastigotes were determined as 8.7, 9.4, 10.9, 25.7 and 25.8 microg/ml respectively. In vivo results of Haplophyllum myrtifolium acidified extract showed that this plant has a limited effect on decreasing the lesion size of experimental mice infected with Leishmania tropica. To the best of our knowledge, this is the first time both the in vitro and in vivo antileishmanial activity of Haplophyllum mrytifolium have been reported in the same research.