Comparison of miRNA profiles in the immune response of pediatric acute appendicitis and pediatric enterobiasis patients caused by Enterobius vermicularis.

BACKGROUND: The aim of this study was to determine and compare the miRNA profile in the immune response with the parasite in pediatric patients with acute appendicitis caused by Enterobius vermicularis and in pediatric patients with enterobiasis. METHODS: A total of 30 tissue samples, which were operated with the diagnosis of pediatric acute appendicitis in the last 10 y and Enterobius vermicularis was detected by histopathological findings, were analyzed. In addition, blood samples were taken from 30 pediatric patients diagnosed with enterobiasis for this study. The miRNAs that activate T and B cells were evaluated by a quantitative real-time PCR, statistically calculated within ΔΔCt values, and fold changes were evaluated by Welch's T test, in which p<0.5 was considered to be significant. RESULTS: It was found that 48 out of 136 (35.3%) miRNAs differed between the pediatric patient and healthy control groups. It was determined that 22 (57.9%) of the different miRNAs were T cell activating miRNAs and 26 (68.4%) were B cell activating miRNAs. While there was a significant difference in miRNA values activating T cells in two patient groups (p<0.01), there was no significant difference in miRNA values activating B cells (p>0.01). CONCLUSIONS: In the study, although Enterobius vermicularis was the causative agent in both patient groups, it was revealed that the immune response of patients with acute appendicitis was more affected than enterobiasis patients.

[Isolation of Cystic Echinococcosis Causative Agents of Echinococcus equinus and Echinococcus ortleppi Species from Humans in the Central Anatolia Region of Türkiye]. / Türkiye'nin Iç Anadolu Bölgesindeki Insanlardan Kistik Ekinokokkoz Etkeni Echinococcus equinus ve Echinococcus ortleppi Türlerinin Izole Edilmesi.

Cystic Echinococcosis (CE) caused by Echinococcus granulosus sensu strico is neglected in Türkiye despite being one of the common diseases due to agriculture being at the forefront, low socioeconomic status and unhygienic animal slaughter. Considering the morbidity, mortality, and difficulties in treatment, more studies and precautions are needed regarding this disease. In this study, it was aimed to genotype Echinococcus isolated from CE patients in the Central Anatolia region. DNA isolation from tissue samples taken from 60 CE patients was performed using the QIAamp DNA FFPE tissue kit. Cytochrome c oxidase subunit gene region of Echinococcus was targeted and JB3/JB4.5 primers were used for genotyping. Polymerase chain reaction (PCR) products were purified according to the instructions for use of the QIAquick PCR purification kit. PCR products were prepared using the ABI Prism BigDye Terminator V3.1 Cycle sequencing kit and the nucleotide sequences in the samples were evaluated with the ABI 3100 sequencing device. The nucleotide sequences obtained in the study were analyzed using MetaPIGA2, MRBAYES v.3.1.2, phyogenetic analysis using parsimony, Unigen programs, maximum likelihood, Bayesian and parsimony methods. It has been found that 88.4% (53/60) of Echinococcus isolates were E.granulosus s.s. in this study. It has been genotyped as 41.7% (25/60) G1, 30.0% (18/60) G3 and 16.7% (10/60) G2 genotype. It has been determined that 6.6% (4/60) of the other Echinococcus isolates were E.equinus and 5.5% (3/60) were E.ortleppi. It was observed that E.equinus and E.ortleppi were isolated from atypically located cysts and from those living in rural areas. The E.equinus and E.ortleppi species were not found in CE patients living in urban areas. CE cases are common in the Central Anatolia region due to dog and cattle breeding, and the disease agent Echinococcus species vary. Genotyping of Echinococcus species is effective in the development of CE treatment and control strategies. Study results can play an active role in the fight against CE, which has formed the basis of the "one health" approach in the world and in Türkiye in recent years.

Identification of Leishmania tropica from Pediatric Visceral Leishmaniasis in Southern Mediterranean Region of Turkey.

Background and Objective: Protozoa of the genus Leishmania are obligate intracellular parasites, and Leishmania species cause a spectrum of species-specific clinical symptoms known as cutaneous, mucocutaneous, and visceral leishmaniasis. For example, Leishmania major and Leishmania tropica cause cutaneous leishmaniasis, while Leishmania infantum and Leishmania donovani cause visceral leishmaniasis (VL). However, molecular studies in recent years have shown that Leishmania species cause different clinical symptoms. Objectives: Our aim was to evaluate the relationship between the clinical and molecular characterization of leishmania isolates in children with VL defined in Turkey, an intercontinental transitional region. Methods: The clinical diagnosis of VL was confirmed by detecting amastigotes in the bone marrow aspirate and/or the rK39 test and/or molecular methods (genus-specific PCR, Real-Time PCR, ITS1 PCR-RFLP, DNA sequencing). Results: Most of the VL patients were referred from the districts of Adana (53.3%) and others from neighboring provinces; Hatay (16.6%), Osmaniye (3%), Gaziantep (3%), Adiyaman (3%), and 20% case were Syrian immigrants A clinical diagnosis of VL was confirmed in 30 patients with different diagnostic methods. 93% was found positive with microscopic examination, 79.1% with rK39 dipstick test, and 60% with genus-specific PCR assay in clinical samples. The Leishmania isolates were identified as L. infantum (40%), L. donovani (26.7%), and L. tropica (23.3%) using Real-Time PCR, ITS1 PCR-RFLP, and DNA sequencing. There was no cutaneous finding in any case in clinical examination.The most common clinical findings were fever (93.3%) and splenomegaly (90%), followed by hepatomegaly (76.6%). The most common laboratory finding was thrombocytopenia (86.6%), followed by anemia (70%). In addition, hemophagocytic lymphohistiocytosis was detected in bone marrow aspiration in two of our patients. Since pentavalent antimony salts treatment initially failed in four patients, it was necessary to switch to Liposomal-Amphotericin B with treatment success. Conclusions: The presence of L. tropica in VL patients, despite the absence of cutaneous findings in any of the cases, shows that this strain can cause VL, contrary to conventional knowledge. In the Adana province, where this study was carried out, L. infantum from CL cases in previous studies should be taken into account, and visceral spread in CL cases and accompanying cutaneous lesions in VL cases should be investigated in detail.

The Molecular Epidemiology of Cystic and Alveolar Echinococcosis in Southeast Turkey.

BACKGROUND: The migration of humans and climatic and environmental changes cause the emergence of infectious diseases. This study aimed to investigate the changes in the molecular epidemiology of the echinococcosis in the southeast region of Turkey after migrations. METHODS: Overall, 159 tissues samples were taken from suspected cases of echinococcosis at the Kilis State Hospital in the southeast region of Turkey. All of the tissues samples were analyzed using histopathology methods, PCR, Real-time PCR methods, DNA sequencing, and phylogenetic analyses in laboratories. RESULTS: The positivity values of the histopathology, the polymerase chain reaction, and the Real-time PCR methods were found to be 14.5% (23/159),15.7% (25/159), and 16.9% (27/159), respectively. 32.0 % (8/25) E. multilocularis of Echinococcus isolates and 68% (17/25) E. granulosus of Echinococcus isolates were identified using PCR methods. 58.8% (10/17) of the E. granulosus isolates were found to be Genotype 1% and 41.2% (7/17) E. granulosus isolates were found to be Genotype 3. CONCLUSION: Molecular methods play an important role in the epidemiology, treatment, and diagnosis of diseases. Increasing immigration in a geographical area may create social, economic, and health problems in that area. For this reason, epidemiological studies of infectious diseases should be updated in areas with immigration.

MicroRNA profile in immune response of alveolar and cystic echinococcosis patients.

It is known that miRNAs are effective in immune response in the diagnosis and treatment of many infectious diseases. However, the miRNAs profile is unknown in Alveolar and Cystic Echinococcosis which can be fatal if left untreated. The miRNAs profile that activates the T and B cells forming the immune system in Alveolar and Cystic Echinococcosis patients was investigated in this study. A total of 50 liver tissue samples were obtained from Alveolar and Cystic Echinococcosis patients in Kilis State Hospital Pathology Laboratory in southeast of Turkey. The circulating cell-free miRNAs were evaluated by a quantitative real-time polymerase chain reaction, statistically calculated within ΔΔCt values and fold changes were evaluated by Welch T test, in which P < .05 was considered to be significant. Twenty-five microRNAs, including let-7a-5p, let-7c, let-7e-5p, miR-15b-5p, miR16, miR-17-5p, miR-23a-5p, miR-24-3p, miR-25-3p, miR-26a-3p, miR-26b-3p, miR-29b-3p, miR-29c-3p, miR-30a-5p, miR-30b-5p, miR-30c-5p, miR-30d-5p, miR-30e-5p, miR-98-5p, miR-101-3p, miR-106b-5p, miR-125b-5p, miR-142-5p, miR-222-3p and miR-223-3p, were found as down-regulated in Alveolar and Cystic Echinococcosis patients than control groups. Twelve miRNAs, including miR-15a-5p, miR-21-5p, miR-27a-3p, miR-29a-3p, miR-146a-5p, miR-181a-5p, miR-181b-5p, miR-181d, miR-181c-5p, miR-195-5p, miR-214-3p and miR-365-3p, were found as up-regulated in Alveolar and Cystic Echinococcosis patients than healthy person. It has been shown that T- and B-cell activities are related in the progressive of both Alveolar and Cystic Echinococcosis in this study. The miRNA panel activated by T and B cells may be important for exploring the mechanisms underlying early development in Alveolar and Cystic Echinococcosis providing novel information that may be used to discover new therapeutics for these diseases.

The increase in neglected cutaneous leishmaniasis in Gaziantep province of Turkey after mass human migration.

Outbreaks of Cutaneous Leishmaniasis (CL) due to war-related factors have been reported in different areas in Turkey and Syria. CL has become the most serious of the infectious diseases which have been reported in Gaziantep in southeast Turkey, during the last three years due to the influx of Syrian refugees. The present research involves an analytical cross-sectional epidemiological study of CL cases diagnosed in the Gaziantep Leishmaniasis Diagnosis and Treatment Center. The patient demographic data, the location of the lesions, the number of the lesions, the duration of the lesions, and the treatment of the lesions are included. The diagnosis of CL was made by microscopic examination of smears in all cases, and 81.1% (900/1110) of which were found to be positive. Out of 900 CL patients, 93.8% (845/900) were Syrian citizens and 6.2% (55/900) were Turkish citizens. The disease was more frequent in females with 53.5% (482/900) and in the age group between 0-20 years with 68.3% (615/900). Distribution of lesions in the body showed that the face was the most affected location with 37% (333/900), and the generation time of lesions was 0-6 months with 71.2% (641/900). 94.7% (852/900) of the CL patients healed without relapse, and 5.3% (48/900) of the CL patients relapsed. CL patients have re-emerged in Gaziantep, located in the southeast of Turkey, as a result of Syrian refugees. The increase in CL frequency is alarming and requires control and prevention measures in highly infected areas including this region.

[Isolation of Balamuthia mandriallaris parasite from air conditioning of the houses in south and southeast of Turkey]. / Türkiye'nin güney ve güneydogusundaki evlerin iklimlendirmelerinde Balamuthia mandrillaris parazitinin izolasyonu.

The free living amoebae cause various infections such as Acanthamoeba keratitis, granulomatous amoebic encephalitis, primer amoebic meningoencephalitis in humans and animals. The free living amoebae Acanthamoeba, Balamuthia mandriallis, Naegleria fowleria and Sappinia species that cause disease in humans have been isolated from many environmental materials until today. However, no isolation has been reported from the filters of the air conditions from the houses used for ventilation. The aim of this study was toinvestigate the existence of free living amoebae using molecular methods in the filters of air-conditions used in the study living area of the people. A total of 30 dust samples were taken from the filtersof air-conditions in Adana and Gaziantep province of Turkey. DNA isolation of the dust samples was performed using the DNeasy PowerSoil kit (Qiagen, Germany) and polymerase chain reaction was done with specific primers of Acanthamoeba spp., B.mandriallis, N.fowleria and Sappinia species. As a result of this study, Acanthamoeba spp. was determined as 33.3% (5/15) and B.mandriallis was determined as6.6% (1/15) in Adana province. On the other hand, Acanthamoeba species was determined as 26.6% (4/15) and B.mandriallis was determined as 13.3% (2/15) in Gaziantep province. N.fowleria and Sappina species were not detectedin both of the cities. DNA sequence analysis was performed for the confirmation of the species and 99% of the results were similar to the other species in GenBank. The rates of Acanthamoeba castellanii and Acanthamoeba griffinii (T3) were determined as %66.6 (6/9) and 33.3% (3/9), respectively by DNA sequencing. Distribution of Acanthamoeba species according to the cities were 33.3% (3/9) for A.castellanii and 22.2% (2/9) for A.griffini in Adana. It was 33.3% (3/9) for A.castellanii and 11.1% (1/9) for A.griffini in Gaziantep. There was no significant difference in the distribution of the parasite species among cities (p> 0.1). It is important to raise awareness of the diseases caused by free living amoebae among people. Acanthamoeba species have been reported frequently from environmental materials in Turkey, but B.mandriallis has not been reported from any environmental sample since this study. The presence of B.mandriallis has been reported in the air-conditions of houses in this study. This result shows that people have risk in terms of illness of free living amoebae in living areas. Our study emphasized that firstly the health personnel and then the people should be informed about the deadly parasites and the cleaning of the air conditions should be done in certain periods.

The epidemic typhus and trench fever are risk for public health due to increased migration in southeast of Turkey.

Pediculus humanus capitis is a small ectoparasitic insect that has lived and feds on human beings for thousands of years. Molecular techniques have been used for Pediculus species identification and evolutionary, phylogenic, and ecological studies. A total of 23 adults of P. h. capitis were collected in Gaziantep, located in southeast Turkey, and DNA was isolated from all P. h. capitis using DNA extraction kit. All DNA samples were screened for investigate of Ricettsia prowazekii, Bartonella quintana and Borrelia recurrentis with real-time polymerase chain reaction. In addition, we investigated genetic variation in DNA samples of Pediculus humanus capitis using the cytochrome oxidase I genetic DNA sequence. We found 4 (17.4%) Ricettsia prowazekii and 3 (13.1%) Bartonella quintana in DNA samples of Pediculus humanus capitis, while we did not find any Bartonella recurrentis in any of the DNA samples. We demonstrated 1.8% genetic variations in DNA samples of Pediculus humanus capitis with Bartonella quintana. The phylogenetic tree based on the cytochrome oxidase I gene revealed that P. h. capitis in southeast Turkey are classified into two clades (clade A, clade B) and Bartonella quintana was found in only clade B. However, we did not find any genetic variations in other DNA samples in this region. The genetic variations may be related to P. h.capitis vector of Bartonella quintana has found in this study. In addition, this study was shown that P. h. capitis do transmit Rickettsia prowazekii and Bartonella quintana to people, epidemic typhus and trench fever may emergence in Gaziantep southeast of Turkey in the future.

Subtype analysis of Blastocystis isolates using SSU rRNA-DNA sequencing in rural and urban population in southern Turkey.

Blastocystis is a common and emerging parasite often seen in many studies conducted in urban population, with scanty reports on rural communities. However, little is known about the public health significance of Blastocystis infection. A total of 28 Blastocystis isolates from 17 (17/28, 60.71%) patients living in rural area and 11 (11/28, 39.29%) patients living in urban area were screened with seven kinds of sequenced-tagged site primers for identification of subtype. PCR products were sequenced with same combination of primers using the BigDye Terminator V 3.1 cycle sequencing kit, as per the manufacturers' protocol on the 3730 DNA analyzer (Applied Biosystems, Carlsbad CA, USA). The cross-comparison of the Blastocystis sequences of samples were determined by the neighbor-joining method based on a distance matrix between sequence pairs to generate dendograms. The following subtypes were identified; subtype 1 (10/28, 35.7%), subtype 3 (7/28, 25.0%), subtype 2 (5/28, 17.8%), subtype 4 (3/28, 10.7%), subtype 5 (1/28, 3.6%), subtype 6 (1/28, 3.6%), and subtype 7 (1/28, 3.6%) in all DNA samples. The comparison of Blastocystis subtypes distribution among the patients from rural and urban area revealed subtype 5 (1/17, 5.9%), subtype 6 (1/17, 5.9%) and subtype 7 (1/17, 5.9%) from patients of rural area but not any of these subtypes in patients living urban area. This study is the first large-scale study to examine the occurrence of Blastocystis in Turkey to shed lights on the cosmopolitan distribution of Blastocystis subtypes in southern part of Turkey. Subtype 5, subtype 6 and subtype 7 were determined in only rural area. The findings of this study suggest that Blastocystis is transmitted from animal to human and possess a zoonotic potential.

Pediculosis capitis is a growing neglected infestation due to migration in southeast Turkey.

Demographic, socio-economical, and environmental changes affecting prevalence of Pediculosis capitis. The aim of this study was to investigate the prevalence of P. capitis and external factors affecting the distribution of P. capitis. A total of 6004 primary-school students between 5 and 11 years were screened for P. capitis at 28 different primary-schools in Gaziantep, located in southeastern of Turkey, during different two education terms (First education term is in September 2013 to May 2014, second education term is in September 2014 and May 2015). The prevalence of P. capitis was found to be positive 1.5 % (90/6004) and 6.9 % (415/6004) in first education term and in second education term, respectively. In this study shown that the rate of P. capitis's prevalence was increased 5.4 % in Gaziantep. P. capitis is a neglected infestation and it has re-emerged in Gaziantep, located in the southeastern of Turkey. Health staff member must improve health education programs in primary-school students especially girl students.

A comparative analysis of different molecular targets using PCR for diagnosis of old world leishmaniasis.

The different sensitivity values were obtained in each study conducted for the diagnosis of leishmaniasis with the polymerase chain reaction (PCR). However, a standardized PCR target for the diagnosis of leishmaniasis does not exist. The aim of the current study, the most ideal PCR target was determined for diagnosis of leishmaniasis. A total of 72 smear and 48 bone marrow samples were analyzed with six different molecular targets to determine their potential as a tool for the specific molecular diagnosis of leishmaniasis using PCR. The positivity-negativity value and the sensitivity-specificity of each PCR targets were calculated. The positivity value of PCR targets were sequenced in different levels in the diagnosis of leishmaniasis from highest to lowest in the order of kDNA-PCR > SSU rRNA-PCR > ITS2-PCR > ITS1-PCR > ME-PCR > HSP70-PCR. The sensitivities of PCR targets except ITS1-PCR, ME-PCR and HSP70-PCR were found to be 100% in cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL) cases as compared to microscopic examination accepted as a gold standard. The sensitivities of ITS1-PCR, ME-PCR and HSP70-PCR were found 96.6%, 90.0% and 86.6%, respectively, in CL-cases. In addition, the sensitivities of ITS1-PCR, ME-PCR and HSP70-PCR were found 90.0%, 70.0% and 60.0%, respectively, in VL-cases. The kDNA genomic region was the most sensitive for routine diagnosis of leishmaniasis. ITS1-PCR restriction fragment length polymorphism, the alternative method for the identification of Old World Leishmania species, did not require culturing of the parasites.

The role of domestic tap water on Acanthamoeba keratitis in non-contact lens wearers and validation of laboratory methods.

Acanthamoeba is increasingly recognized as an important cause of keratitis in non-contact lens wearers while contact lens wear is the leading risk factor for Acanthamoeba keratitis (AK). It is unlikely that the Acanthamoeba colonization is a feature which is effective only in patient's homes with infectious keratitis since the organism has been isolated from domestic tap water. Two hundred and thirty-one (231) corneal scrapings were taken from infectious keratitis cases, and four contact lens solutions and domestic tap waters were taken from 22 out of 44 AK-diagnosed patient's homes. Microscopic examination, culture, PCR, real-time PCR and DNA sequencing analyses were used for AK-diagnosed samples. The real-time PCR was the most sensitive (100 %) one among the methods used in diagnosis of AK. The 44 (19.0 %) out of 231 corneal scrapings, 4/4 (100 %) contact lens solution and 11/22 (50 %) of domestic tap water samples were found to be positive by real-time PCR for Acanthamoeba. A. griffini (T3), A. castellanii (T4) and A. jacobsi (T15) genotypes were obtained from corneal scrapings, contact lens solutions and domestic tap water samples taken from the patient's homes diagnosed with AK. The isolation of Acanthamoeba containing 6/22 (27.3 %) A. griffini (T3), 14/22 (63.6 %) A. castellanii (T4) and 2/22 (9.1 %) A. jacobsi (T15) from the domestic tap water outlets of 22 of 44 (50 %) of patient's homes revealed that is a significant source of these organisms. A. griffini (T3) and A. jacobsi (T15) genotypes have not been determined from AK cases in Turkey previously. Thus, we conclude that Acanthamoeba keratitis is associated with exposition of patients who has ocular trauma or ocular surface disease to domestic tap water in endemic or potentially endemic countries.

Two different myiasis cases in southeast of Turkey: ophthalmomyiasis and cutaneous myiasis.

Myiasis has become increasingly prevalent, particularly when human activity is carried out in environments with poor hygiene. We reported two cases of human myiasis in this paper. All of myiasis cases initially presenting to the Emergency Department of Kilis State Hospital in Turkey were identified. We present one case of ophthalmomyiasis caused by Oestrus ovis and one case of cutaneous myiasis caused by Lucilia sericata. Both of the myiasis cases were reported from rural area case limiting the exposure to adult flies and exterminating the flies will play important role in preventing the myiasis.

Clinical manifestations and genetic variation of Leishmania infantum and Leishmania tropica in Southern Turkey.

L. infantum was isolated from cutaneous leishmaniasis (CL) skin lesions in patients having no signs and symptoms of visceral leishmaniasis (VL). Similarly, L. tropica had previously been isolated from patients with VL in the absence of cutaneous lesions. It was not certain how visceralization occurred. Smears (207) and bone marrow samples (135) were taken from CL and VL-suspected patients, respectively. Microscopic examination, ITS1-PCR, RFLP and DNA sequencing for all samples were analyzed. The microscopic examination of smears was found to be 61.3% (127/207) in CL-suspected cases and bone marrow samples were found to be positive 8.8% (12/135) in VL-suspected cases. L. tropica 48.6% (72/148), L. infantum 35.8% (53/148), L. major 15.6% (23/148) in CL, and L. infantum 56.3% (18/32), L. donovani 31.2% (10/32), L. tropica 12.5% (4/32) in VL were found with PCR-RFLP. In addition, the DNA sequencing revealed a genetic variation in L. infantum (variants 1-3) and L. tropica (variants 1-5). We assume that the increased disease occurrence may have resulted from geographical expansion of disease, changing patterns of international travel, population migrations, non-immune people into endemic regions of infected people into non-endemic regions. In this study, L. infantum (variant 3) only in CL-patients and L. tropica (variant 2) only in VL-patients were identified. We hypothesize that genetic variation might play a role in the causation of CL and VL in southern Turkey and the genetic variants may differ according to the geographical location among Leishmania strains.

Genotyping of Acanthamoeba T15: the environmental strain in Turkey.

BACKGROUND: Environmental sources are potential sources for the transmission of Acanthamoeba in humans and other mammals. METHODS: A total of 50 water samples from hot springs and swimming pools, and 50 soil samples were taken from Adana, Afyon, Kutahya, Mersin and Nigde provinces in Turkey. Samples were analysed using 18S rRNA-DNA sequencing. RESULTS: Acanthamoeba griffini (T3), Acanthamoeba castellanii (T4) and Acanthamoeba jacobsi (T15) were found in water samples. Acanthamoeba griffini (T3) and Acanthamoeba castellanii (T4) were detected in soil samples. CONCLUSIONS: In Turkey, this was the first time that Acanthamoeba jacobsi (T15) was detected in water samples.

Comparison of clinical samples and methods in chronic cutaneous leishmaniasis.

This study aimed at finding out the most effective clinical samples and methods in chronic cutaneous leishmaniasis (CCL). Smear, aspiration fluid, and filter paper samples were taken from 104 skin lesions of suspected cases with CCL, and they were compared using microscopic examination, culture, and molecular methods. We characterized four different forms of CCL and identified the causative agents in CCL forms using high-resolution melting curve real-time polymerase chain reaction assay. We observed that smear was detected to be the most sensitive (63.5%) among clinical samples, and real-time polymerase chain reaction method was the most sensitive (96.8%) among the methods used in diagnosis of CCL. We identified 68.8% Leishmania tropica and 31.2% L. infantum in papular lesions, 69.2% L. infantum and 30.8% L. tropica in nodular lesions, 57.9% L. tropica and 42.1% L. major in ulcerating plaque lesions, and 55.5% L. tropica and 44.5% L. major in noduloulcerative lesions in CCL patients.

The emergence of Leishmania major and Leishmania donovani in southern Turkey.

BACKGROUND: In southern Turkey, Leishmania tropica and L. infantum are both the causative agents of cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL), respectively. However, L. major and L. donovani were known to exist after the influx of Syrian refugees. METHODS: Between the years of July 2003 and July 2013, a total of 167 smears and 113 bone marrow samples were taken from CL and VL-suspected cases, respectively. Samples were analysed through real-time PCR and ITS1 DNA sequencing. RESULTS: One hundred and seven 64% (107/167) smears and 56% (63/113) bone marrow samples were positive for leishmaniasis according to the real-time PCR. Three different Leishmania species were found in the 107 CL cases by real-time PCR: 42% (45/107) L. tropica, 36.5% (39/107) L. infantum and 21.5% (23/107) L. major. In addition, three different Leishmania species were identified in the 63 VL cases: 60.3% (38/63) L. infantum, 30.2% (19/63) L. donovani and 9.5% (6/63) L. tropica using real-time PCR. The results of real-time PCR were confirmed with Leishmania ITS1 DNA sequencing. CONCLUSIONS: This study revealed that in southern Turkey, L. major and L. donovani were the aetiological agents of CL and VL, respectively. It was assumed that emergence of L. major and L. donovani was due to influx of Syrian refugees, as well as the effects of global warming.

Non-contact lens use-related Acanthamoeba keratitis in southern Turkey: evaluation of risk factors and clinical features.

PURPOSE: To assess the diagnostic methods, risk factors, and clinical features of Acanthamoeba keratitis cases in patients who do not wear contact lenses. METHODS: Medical records of 26 consecutive patients with non-contact lens-related Acanthamoeba keratitis, who were followed up at the tertiary eye care center between May 2010 and May 2012, were analyzed. Laboratory, demographic, and clinical findings were evaluated pertaining to the patients. RESULTS: Twenty-six non-contact lens-related Acanthamoeba keratitis cases were included in the study. The main risk factors were trauma (group 1, n = 13 patients) and ocular surface disease (group 2, n = 12 patients). One patient had both of the risk factors mentioned above. Overall test results showed that Acanthamoeba positivity rates were 15.3% for direct microscopy, 46.1% for culture, 92.3% for conventional polymerase chain reaction (PCR), and 100% for real-time PCR. The rates of full-thickness corneal involvement and ring-shaped infiltrations were higher in group 2, whereas superficial keratitis and radial keratoneuritis were higher in group 1. The final visual acuities were significantly better in group 1 than group 2 (p<0.025). CONCLUSIONS: This study is the first regional report from Turkey about Acanthamoeba keratitis in non-contact lens users. A majority of cases admitted to a tertiary eye care center were related to trauma or ocular surface disease. Physician suspicion is critically important for the timely diagnosis of these cases. At this point, molecular diagnostic tests (PCR or real-time PCR) seem to support the clinical diagnosis of Acanthamoeba keratitis with the help of fast and reliable results.

Evaluation of the transmission mode of B. hominis by using PCR method.

Blastocystis hominis is a common intestinal parasite observed in fecal examination. On the other hand, the transmission of this parasite is certainly unknown. The transmission of B. hominis can be realized by animal contact and the contamination by water and food with excreted cysts from the reservoir hosts. B. hominis isolated from 25 humans, their pets, and tap water was identified by polymerase chain reaction using sequenced tag site primers in this study. B. hominis isolates obtained from humans and pets were identified as subtype1, subtype2, and subtype3 while B. hominis isolates obtained from tap water were also identified as subtype1. The B. hominis isolates obtained from humans in this study were defined as the same as the subtypes of the B. hominis isolates obtained from the pets, of which these people keep at their homes, and the tap water. These findings reveal that the source of B. hominis infection could be pets and tap water.

Detection of Plasmodium vivax by nested PCR and real-time PCR.

Malaria is endemic in the Cukurova region while it is sporadic in other regions of Turkey. Therefore, the laboratory and clinical diagnosis of malaria is important for the treatment of malaria. In this study, 92 blood samples that were taken from the suspected malaria patients for routine diagnosis in a period of 10 years between 1999 and 2009 were analyzed. All of these blood samples were examined by microscopic examinations using Giemsa-stained thick blood films, nested PCR, and real-time PCR. The sensitivity-specificity and positive-negative predictive values for these diagnostic tests were then calculated. It was found that the positive predictive values of microscopic examination of thick blood films, nested PCR, and real-time PCR were 47.8%, 56.5%, and 60.9% for malaria, respectively. The real-time PCR was found to have a specificity of 75% and sensitivity of 100%, while specificity and sensitivity of nested PCR was found 81.2% and 97.7% according to the microscopic examination of thick blood films, respectively.