[Investigation of Leishmania RNA Virus 2 in Leishmania major and Leishmania tropica Strains Isolated from Cutaneous Leishmaniasis Patients in Türkiye]. / Türkiye'deki Kutanöz Leysmanyazis Hastalarindan Izole Edilen Leishmania major ve Leishmania tropica Izolatlarinda Leishmania RNA Virüsü 2'nin Arastirilmasi.

Leishmania RNA virus (LRV) is a double-stranded RNA (dsRNA) virus that is thought to contribute to the severe inflammatory response of the causative Leishmania parasite in the mammalian host by being present in many isolates of Leishmania spp. In our study, it was aimed to obtain data on the presence of Leishmania RNA Virus 2 (LRV2), which is thought to cause a change in the clinical course of leishmaniasis, in Leishmania major and Leishmania tropica isolates isolated from cutaneous leishmaniasis (CL) patients in Türkiye. Leishmania strains stored in liquid nitrogen tank by cryopreservation in Manisa Celal Bayar University Faculty of Medicine Parasite Bank were resuscitated under suitable conditions and cultivated in NNN and RPMI-1640 media. Then, the isolates were allowed to enter the logarithmic phase in a 26ºC incubator and DNA isolations were made using the "High Pure PCR Template Preparation Kit". Real-time polymerase chain reaction (Rt-PCR) melting analyzes were applied to the DNAs obtained by using primers and probes specific to the internal transcribed spacer-1 (ITS-1) gene region of Leishmania. After RNA isolation from promastigote suspension, cDNA synthesis was performed by reverse transcription. After gel electrophoresis with PCR amplification products, dsRNA band formation was evaluated in terms of LRV2 positivity under ultraviolet light. Among the 20 examined Leishmania spp. isolates (10 L.tropica and 10 L.major), four (three L.tropica, one L.major) were found to be positive for the presence of LRV2. Although the mechanism of LRV in recent studies has not been fully understood, it is known that it exacerbates the clinic of the disease and even has an effect on the formation of drug resistance by the parasite. It is important to obtain data on the presence of LRV in our country and to contribute to various clinical, drug development, prevalence studies, diagnosis and treatment of the disease in the future.

Determination of sand fly fauna and molecular detection of Leishmania in sand flies in Antalya Province, Southern Turkey.

Visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL) are diseases transmitted by infected female sand flies. Since the eradication of malaria in Turkey, CL is the main vector-borne disease in the country, with more than 2000 cases per year, making it a significant public health problem. The aims of this study were to carry out an entomological survey in Antalya Province, an endemic area for CL in the Mediterranean Region of Turkey, to identify sand fly fauna and to screen female specimens for the presence of Leishmania parasites (Leishmania infantum, L. tropica, L. major, and L. donovani) using molecular analysis. Sand flies were collected in 42 localities of seven districts in Antalya Province using CDC miniature light traps in two different periods, June 2012 and September 2013. The specimens were kept in 96% ethanol until the dissection was done. The head and genitalia of the specimens were cut for preparing individual slides for species identification. The rest of the body of female specimens was kept separately. The specimens were identified at the species level, and 27 pools were generated according to the locations and species for screening the presence of Leishmania. A commercial kit was used for DNA extractions. Real-time and conventional polymerase chain reaction (PCR) targeting the internal transcribed spacer region (ITS1) were then performed. In total, 1306 specimens comprising nine species belonging to the Phlebotomus genus were collected in the study region, with Phlebotomus neglectus/syriacus (38.82%) the most abundant, followed by P. alexandri (21.67%) and P. tobbi (20.44%). In the 27 pools, Leishmania infantum DNA was detected in four pools containing P. neglectus/syriacus and one pool containing P. tobbi. In conclusion, the sand fly fauna in the Antalya Province is diverse. The probable vector sand fly species are P. neglectus/syriacus and P. tobbi with high dominance (59.26%), which indicates a high risk of CL transmission. The data presented here may help to shed more light on the transmission cycles of the Leishmania parasite in this CL endemic area.

The Toxicity of Essential Oils From Three Origanum Species Against Head Louse, Pediculus humanus capitis.

PURPOSE: Pediculosis caused by human head louse, Pediculus humanus capitis De Geer (Anoplura: Pediculidae) is a public health problem with a high prevalence, especially among children in school age. Different chemical pediculicides (insecticides) are used against lice but the repeated application of these pediculicides for the treatment of P. humanus capitis infestation in the past decades has resulted in the development of resistance. The plant essential oils have been used as alternative sources of lice treatment agents. Pediculicidal activity of the essential oils of three Origanum species (O. onites, O. majorana and O. minutiflorum) against P. humanus capitis adults was examined in this study. METHODS: Essential oils from aerial parts of three Origanum species were obtained by hydrodistillation using a Clevenger-type apparatus. P. humanus capitis specimens used were obtained from infested children by combing method at primary schools. Adult lice were exposed to three dilutions (1%, 0.5% and 0.1% w/v) of tested oils for 5 min using the adult immersion test. RESULTS: According to the results, all Origanum essential oils at 1% concentration have been shown to significantly decrease the rate of limb, bowel and abdomen movements, producing more than 90% mortality after 12 h. CONCLUSION: Our results indicated that Origanum essential oils can be used for the development of new pediculicides against head louse.

Entomological Survey for the Detection of Sand Fly Fauna and Vector Species in the Cutaneous Leishmaniasis Endemic Area in East Mediterranean Region of Turkey, Mersin Province.

Cutaneous (CL) and visceral (VL) forms of leishmaniasis, transmitted by sand flies, are seen in all countries located in Mediterranean Basin including Turkey. In this study, we aimed to conduct an entomological survey for the detection of sand fly fauna and vector species in Mersin province, one of the important endemic areas for CL in Turkey. In total, 912 sand fly specimens were collected in 2010 and 2011 using CDC light traps. Nine Phlebotomus (Diptera: Psychodidae) and three Sergentomyia (Diptera: Psychodidae) species were detected. Of the collected Phlebotomus sand flies, P. sergenti Parrot, 1917 (30.1%) was the most dominant followed by P. alexandri Sinton, 1928 (18.2%), P. neglectus/syriacus Tonnoir Adler (12.0%), P. tobbi Adler & Theodor, 1930 (11.7%), and P. papatasi Scopoli, 1786 (10.2%), while S. minuta Rondani, 1843 (11.3%) was the dominant species among Sergentomyia. During the field work in 2011, female specimens (n = 81) were screened for the presence of Leishmania promastigotes by midgut dissection, and all were found negative. The rest of the collected female specimens (n = 334) were pooled according to species (P. alexandri, P. neglectus/syriacus, P. papatasi, P. sergenti, P. simici, and P. tobbi) and location (Mut, Silifke, and Anamur). In total, 29 pools were generated and real-time ITS1 PCR assay was performed to detect and identify natural Leishmania Ross, 1903 (Kinetoplastida: Trypanosomatida) infection. Two pools, both from Mut town, containing P. sergenti specimens were found positive and Leishmania tropica Ross, 1903 was identified as an infectious agent for both pools. In conclusion, the sand fly fauna was determined in an endemic area for CL. The detection of L. tropica DNA in P. sergenti specimens showed the possible vectorial role of this species in Mersin province.

Detection of Leishmania major and Leishmania tropica in domestic cats in the Ege Region of Turkey.

Leishmaniosis is a group of diseases caused by different species of Leishmania parasites in mammalian species. The aim of the present study was to investigate the presence of Leishmania spp. DNA in cats using real time polymerase chain reaction (RT-PCR) assays targeting internal transcribed spacer (ITS1) and heat-shock protein 70 gene (Hsp70) regions with Leishmania species-specific primers and probes. Blood samples were collected from 147 cats (73 female; 74 male) in the endemic regions for zoonotic visceral leishmaniasis in the western provinces of Turkey and analyzed using two RT-PCR assays. Additionally, Hsp70 RT-PCR products were sequenced. ELISA assays for feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) were also carried out for 145 of the 147 samples. Overall, 13/147 (8.84%) cats were positive for Leishmania by RT-PCR (4 L. major and 9 L. tropica). FIV and FeLV antibody and/or antigen was detected in 4 and 5 cats among Leishmania DNA positives, respectively. To the best of our knowledge, this study is the first to investigate and report the presence of L. major and L. tropica infections in a large group of domestic cats in Turkey. The results obtained indicate that species identification of Leishmania is essential for epidemiological understanding and that clinical signs alone are not indicative for leishmaniosis in cats, as it is in dogs. This study suggests that extensive research should be carried out in cat populations in order to fully understand the role of cats in the epidemiology of the disease.

Seroepidemiological and entomological survey in a new focus of zoonotic visceral leishmaniasis in Kars province, Northeastern Turkey.

Visceral leishmaniasis (VL) has now been recorded from 38 provinces of Turkey. Twenty-one VL cases were reported within six years from settlements located in most northeastern Turkey and we therefore aimed to carry out an entomological and seroepidemiological survey in this new focus for clarifying risk factors. Blood samples from 290 children and 165 dogs were collected. Sera samples were investigated for anti-Leishmania antibodies using indirect fluorescent antibody test. Sand fly collection for determining the fauna and seasonal activity was performed in all settlements by CDC light traps between June and September 2006. Although no seropositive child was detected during the survey the overall seroprevalence rate of canine leishmaniasis was found as 7.2%. A total of 4154 sand flies were collected and 10 species of genus Phlebotomus were identified belonging to Adlerius, Larroussius, Paraphlebotomus and Phlebotomus subgenera. Among them Phlebotomus kandelakii s.l. (55.44%), Phlebotomus balcanicus (12.62%) and Phlebotomus neglectus (4.40%) was detected as probable vector species for this new focus. The poor sanitation, very high population size of sand flies, probably because of very short season, no control measures for sand flies as well as dogs, and presence of microclimate suitable for sand flies were considered as main risk factors in the area.

Treatment of head lice (Pediculus humanus capitis) infestation: is regular combing alone with a special detection comb effective at all levels?

Head lice infestation (HLI) caused by Pediculus humanus capitis has been a public health problem worldwide. Specially designed combs are used to identify head lice, while anti-lice products are applied on the scalp for treatment. In the present study, we aimed to test whether combing only by precision detection comb (PDC) or metal pin comb (MPC) could be effective alternatives to the use of anti-lice products in children. A total of 560 children from two rural schools in Turkey were screened. In the PDC trial, children were combed every second day for 14 days, while in the MPC trial, combing was performed once in every four days for 15 days. Children were divided into two groups (dry combing and wet combing) for both trials and results were compared. The results showed no significant differences between dry and wet combing strategies for both combs for the removal of head lice (p > 0.05). The number of adult head lice declined significantly on each subsequent combing day in both approaches, except on day 15 in the MPC trial. In the end, no louse was found in 54.1 and 48.9% of children in the PDC and MPC trials, respectively. Since family members of infested children were not available, they were not checked for HLI. Four times combing within 2 weeks with MPC combs was found effective for both treatment of low HLI and prevention of heavy HLI. In conclusion, regular combing by special combs decreases HLI level in children and is safely applicable as long-term treatment.

Comparing the Efficacy of Commercially Available Insecticide and Dimeticone based Solutions on Head Lice, Pediculus capitis: in vitro Trials.

OBJECTIVE: Head lice infestation is a public health and social problem for almost all countries worldwide. For its treatment, insecticide and dimeticone-based solutions are currently available in the markets in many countries. We aimed to compare the efficacy of commercially available anti-head lice shampoos containing insecticide and physically effective products with different percentages of dimeticone using an in vitro technique. METHODS: Head lice specimens were collected from primary school children using special plastic and metal combs. Anti-head lice products were commercially purchased and used directly. The specimens were placed one by one in 5-cm Petri dishes containing a slightly wet filter paper and were kept in a plastic cage at 28±2°C and 50%±20% relative humidity. A standardized protocol was used for testing all the products, and mortality data were obtained after 24 h. Two control tests were performed with each batch of trials. For each product and control, 10-20 head lice specimens were used, and the results were statistically analyzed. RESULTS: Our study demonstrated that among all the tested products, two products containing mineral oils [5.5% dimeticone & silicone (patented product) and dimeticone (no percentage mentioned in the prospectus) & cyclopentasiloxane] were found to be more effective for killing head lice in vitro. CONCLUSION: Physically effective products can be repetitively used because they are non-toxic and resistance to them is not expected. To control the infestation at a public level, the use of these products needs to be encouraged with respect to their cost price.

[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.

Investigation of in vitro antileishmanial activity of moxifloxacin, linezolid and caspofungin on Leishmania tropica promastigotes.

OBJECTIVE: This study aimed to evaluate the potential in vitro anti-leishmanial activities of moxifloxacin, linezolid and caspofungin against Leishmania tropica. METHODS: In vitro effects of all agents were studied by using the microdilution method. For this purpose, serial dilutions of the aforementioned agents were prepared in concentrations between 4096 µg/mL-0.008 µg/mL. Afterwards, promastigotes incubated in suitable medium were counted with the hemocytometer and adjusted as having a last concentration of 2.5 x 10(6) cells/mL in wells containing medium+antibiotic or antifungal. After incubation live promastigotes were counted with the hemocytometer and inhibitor concentrations (IC(50)) were determined by comparing with the control that contained no antibiotics or antifungal. RESULTS: IC(50) values of moxiloxacin, linezolid and caspofungin were found as 194.7 µg/mL, 896 µg/mL and 235.7 µg/mL, respectively. CONCLUSION: As a result, moxifloxacin was found to be effective in lower concentrations than the other studied agents against L. tropica promastigotes.

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.

Treatment of head lice with dimeticone 4% lotion: comparison of two formulations in a randomised controlled trial in rural Turkey.

BACKGROUND: Dimeticone 4% lotion was shown to be an effective treatment for head louse infestation in two randomised controlled trials in England. It is not affected by insecticide resistance but efficacy obtained (70-75%) was lower than expected. This study was designed to evaluate efficacy of dimeticone 4% lotion in a geographically, socially, and culturally different setting, in rural Turkey and, in order to achieve blinding, it was compared with a potential alternative formulation. METHODS: Children from two village schools were screened for head lice by detection combing. All infested students and family members could participate, giving access to treatment for the whole community. Two investigator applied treatments were given 7 days apart. Outcome was assessed by detection combing three times between treatments and twice the week following second treatment. RESULTS: In the intention to treat group 35/36 treated using dimeticone 4% had no lice after the second treatment but there were two protocol violators giving 91.7% treatment success. The alternative product gave 30/36 (83.3%) treatment success, a difference of 8.4% (95% CI -9.8% to 26.2%). The cure rates per-protocol were 33/34 (97.1%) and 30/35 (85.7%) respectively. We were unable to find any newly emerged louse nymphs on 77.8% of dimeticone 4% treated participants or on 66.7% of those treated with the alternative formulation. No adverse events were identified. CONCLUSION: Our results confirm the efficacy of dimeticone 4% lotion against lice and eggs and we found no detectable difference between this product and dimeticone 4% lotion with nerolidol 2% added. We believe that the high cure rate was related to the lower intensity of infestation in Turkey, together with the level of community engagement, compared with previous studies in the UK. TRIAL REGISTRATION: Current Controlled Trials ISRCTN10431107.

[The serological and entomological survey of zoonotic visceral leishmaniasis in Ayvacik Region of Canakkale Province, Turkey]. / Canakkale Ili Ayvacik Bölgesinde Zoonotik Visseral Leishmaniasisin Serolojik ve Entomolojik Olarak Arastirilmasi.

The field studies were carried out in Ilyasfaki village belonging to Ayvacik town, Kalabakli village and center of Kepez town belonging to Canakkale province to determine the epidemiological situation of visceral leishmaniasis between June and August 2007. A total of 27 blood samples were taken from the dogs which are known as reservoir of Leishmania infantum causing visceral leishmaniasis in human and dogs. The dogs were also examined physically for clinical symptoms of the disease. A total of 789 sand fly specimens were collected using CDC miniature light traps and examined. Six Phlebotomus and one Sergentomyia species were identified as: Phlebotomus negletus, P. tobbi, P. simici, P. papatisi, P. perfiliewi, P. halepensis and S. theodori. Among these species P. neglectus, was found to be a dominant species in Ilyasfaki village by the rate of 94.4%, while P. tobbi was also found to be a dominant species in city center (including Kepez) and Kalabaki village by the rate of 50% and 48.1%, respectively. No seropositive dogs were found by IFA test. Seropositivity below cut-off level was only observed in two dogs from Kepez, in dilutions of 1/16 and 1/64. The risk level for human and canine visceral is found to be high and it requires more field studies on canine leishmaniasis in and around the area.

In vitro activity of Arbutus unedo against Leishmania tropica promastigotes.

Pentavalent antimonials are the first choice for the treatment of anthroponotic cutaneous leishmaniasis (ACL) in health centers in Turkey, however in rural areas, traditional plants may be preferred for the treatment of lesions. In recent years a number of papers are published related to the natural products especially plant derivates. Our aim is to investigate the antileishmanial effect of Arbutus unedo which is a wild plant mainly grown in maquis and rocky places of the seabord in South Europe. In the present study, the ethanolic, water and n-hexane extracts from the leaves of Arbutus unedo were originally tested in vitro against Leishmania tropica promastigotes. The ethanol extract of Arbutus unedo leaves at the concentrations of 100, 250, 500 microg/ml were found to be more effective than the other extracts (p:0.000). Our study showed that the ethanolic extract of Arbutus unedo leaves can be a promising antileishmanial agent and further experiments are needed.

[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.

Comparison of two combs in the detection of head lice in school children.

The diagnostic efficacies of two metal pin lice combs having different spacing between their teeth were compared in two cohorts of school children, aged between 7 and 15 years, from different villages of Manisa, Turkey. Head lice infestation was evaluated with comb A (0.18 mm of distance between the teeth) in 95 children in Cavusoglu village, and with comb B (0.15 mm of distance between the teeth) in 146 children in Yesilköy village, compared to visual inspection. Five of 95 (5.3%) children in Cavusoglu village and 5 of 146 (3.4%) children in Yesilköy village were found to harbor live head lice with combing, while none was detected during the visual inspection. The difference between the infestation rates of two cohorts was statistically insignificant (p > 0.05). With visual screening, 15 and 16 children with louse eggs were identified in Cavusoglu and Yesilköy villages, respectively. With combing in these villages, 5 and 8 children, respectively, with louse eggs were identified and 3 and 5 children, respectively, were detected only with combing, not with visual screening. In conclusion, combing is more effective than visual inspection in head lice detection, but there were no significant difference between the two combs either in lice detection or in usage.

Subungual myiasis in a woman with psychiatric disturbance.

Myiasis is caused by the invasion of tissues or organs of men and animals by dipterous larvae. The disease is infrequent in Turkey. A case of a 65-year-old woman having been initially diagnosed with chronic psychosis, and found to have a left big toe nail invaded by the larvae of Calliphora spp., is presented. A total of 17 maggots were removed from the left big toe of the patient, which were then determined as Calliphora spp. The patient has undergone surgical withdrawal of both nails of her big toes, followed by the administration of oral prophylactic antimicrobial treatment. This is the second recorded case of subungual myiasis by Calliphora spp. in Turkey. Myiasis should be considered in patients with lower personal hygiene, especially with chronic psychiatric disturbances.

[Isosporiasis in an immunocompetent child: case report]. / Bagisiklik sistemi saglam bir çocukta isosporiasis: Olgu Sunumu.

A 12-year old boy was presented as a case report. He had been diagnosed with isosporiasis during a field study, conducted in a village of Manisa province, during an investigation of the incidence of intestinal parasites.

Rural life, lower socioeconomic status and parasitic infections.

This study was conducted both to assess the prevalence of parasitic infections in two neighboring villages of Manisa province in Turkey, with different socioeconomic levels and investigate the influences of some individual and environmental factors on these infections. A total of 100 school children were enrolled and their cellophane tape and stool samples were examined for parasites. Pediculus capitis infestation was diagnosed by visual inspection of the hair. Although the prevalences of both intestinal parasitosis and pediculosis were high in both villages, there was a statistically significant difference for pediculosis, but not for intestinal parasitosis. Lack of social security and father's regular job were found to be highly correlated with all parasitic infections. Pediculosis was more common in younger children, girls and those in crowded families. These results indicate not only the high prevalence of parasitic infections, but also the urgent need for the improvement of basic health services and infrastructure in the rural areas, crowded with poorer immigrant population.