[Evaluation of Pre-analytical Process with Quality Indicators and Six Sigma Methodology in the Parasitology Laboratory of a Tertiary Healthcare Center]. / Üçüncü Basamak Saglik Merkezinin Parazitoloji Laboratuvarinda Analiz Öncesi Sürecin Kalite Belirteçleri ve Alti Sigma Yöntemi ile Degerlendirilmesi.

Laboratories have important role in decisions related to the patient. Laboratory performance needs to be evaluated to ensure accurate and sustainable laboratory results. Total test process consists of pre-analytical, analytical and post-analytical sub-processes. Most of the laboratory errors occur in pre-analytical process, which is mostly outside the laboratory, and this important situation has to be monitored by laboratory specialists. Although the standard statistical methods in which the frequency is evaluated can reveal which error is more than the others, they cannot determine which error is needed due to the absence of accepted target values. The decision to intervene in errors can only be made according to the targets by evaluating with methods such as six-sigma and quality indicators. Six-sigma method; is a quality management tool that provides information about process performance. Low sigma level indicates variability or errors in the relevant process. Quality indicators have been developed to measure quality and efficiency of laboratory processes. Use of quality indicators is effective in reducing errors, increasing patient safety and helping to meet ISO-15189 requirements. In this study, it was aimed to evaluate pre-analytical process performance in Parasitology Direct Diagnosis Laboratory of Ege University Faculty of Medicine according to the quality targets of International Federation of Clinical Chemistry and Laboratory Medicine Working Group on Laboratory Errors and Patient Safety (IFCC WG-LEPS) and the six-sigma method. The data of rejected samples in our laboratory during the period 2014-2017 were obtained retrospectively from laboratory information system. Errors were classified using laboratory errors classification system. Quality indicators were calculated for each error category and assessed according to IFCC WG-LEPS quality targets. Pre-analytical sigma level was calculated for each year. Our pre-analytical process sigma goal was 4.6. Sigma levels were calculated according to the reasons of rejection and Pareto analysis was performed. All of the rejected samples were pre-analytical process errors. Unacceptable quality indicators according to the IFCC WG-LEPS targets were found as "insufficient sample" in 2015; "insufficient sample" and "inappropriate sample tube" in 2016 and 2017. Our pre-analytical process sigma levels according to the rejection reasons were found to be 4.39, 4.31, 4.11, 4.17, respectively in 2014- 2017. "Improper test request" in 2014, and "insufficient sample" in 2015-2017 had sigma levels below 4.6. In addition "improper test request" in 2014, and "insufficient sample" in 2015, 2016 and 2017 were noticeable in Pareto analysis. In this study, pre-analysis process was evaluated with six sigma method and quality indicators and the areas open for improvement were determined quantitatively. We found "insufficient sample", "improper test request" and "inappropriate sample tube" indicators as inappropriate according to our target values with both quality indicators and six-sigma methods. For this reason, we have planned video conference training focused on error sources for all employees. We consider that risk and number of errors will be reduced and efficiency of whole test process can be increased by evaluating pre-analytical process with accepted methods and monitoring the results. Process evaluation studies with six-sigma and quality indicators are limited in microbiology and parasitology laboratories. We think that laboratory quality is indispensable and this study will be an example for the laboratory specialists who want to evaluate pre-analytical process of their laboratories.

[Infecting glial cells with antimony resistant Leishmania tropica: A new ex-vivo model]. / Glia hücrelerinin antimona dirençli Leishmania tropica ile enfekte edilmesi: Yeni bir ex-vivo modeli.

Leishmaniasis is a vector-borne zoonotic disease that shows different clinical features like cutaneous, mucocutaneous, visceral and viscerotropic forms. The protocols used in the treatment of leishmaniasis are toxic and have many limitations during administration. One of the limitations of treatment is the resistance against the protocols in practice. There is also a need to define new treatment options especially for resistant patients. Ex-vivo models using primary cell cultures may be a good source for evaluating new drug options in patients with antimony resistance, in addition to in-vitro and in-vivo studies. In this study, it was aimed to define a new ex-vivo culture model to evaluate treatment options in patients with cutaneous leishmaniasis who did not respond to treatment. In our experimental model of ex-vivo infection, Leishmania tropica promastigotes isolated from a case previously diagnosed with cutaneous leishmaniasis were used. The primary astroglial cell culture used for the ex-vivo model was prepared from 2-3 days old neonatal Sprague Dawley rat brains under sterile conditions by the modification McCarthy's method. The astroglia cells, which reached sufficient density, were infected with antimony resistant L.tropica promastigotes. After 24 hours of incubation, the supernatant on the cells were collected, the cell culture plate was dried at room temperature, then fixed with methyl alcohol and stained with Giemsa to search for L.tropica amastigotes. Amastigotes were intensely observed in glia cells in primary cell cultures infected with L.tropica promastigotes. No promastigotes were seen on Giemsa stained preparations of the precipitates prepared from the bottom sediment after the centrifugation of the liquid medium removed from the infected plates. In this study, promastigotes from a cutaneous leishmaniasis patient unable to respond to pentavalent antimony therapy were shown to infect rat glia cells and converted to amastigote form. This amastigote glial cell model, as far as we know, is the first model in the literature produced by L.tropica. The occurrence of L.tropica amastigote forms in glia cells may be indicative of the ability of Leishmania species to infect the central nervous system. The central nervous system may be an area for the Leishmania amastigotes to escape from the immune system in cases of leishmaniasis without a treatment response. Our study is important because it is the first study to show the infection of glia cells with L.tropica amastigotes.

[Comparison of polymerase chain reaction using kinetoplast DNA specific primers and other parasitological methods in the diagnosis of clinical samples of suspected patients with cutaneous leishmaniasis in Sanliurfa]. / Sanliurfa'da kutanöz leysmanyazis süpheli hastalardan alinan klinik örneklerin tanisinda kinetoplast DNA'sina özgül primerlerin kullanildigi polimeraz zincir reaksiyonu ile diger parazitolojik yöntemlerin karsilastirilmasi.

Visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL) are seen endemically in Turkey and CL caused by Leishmania tropica is an important public health problem in southeastern as well as other regions of Turkey. The diagnosis has been usually made by clinical view of lesion and/or parasitologically using lesion aspiration smears. Histological examination does not, always reveal the parasite in the skin biopsy, particularly in chronic lesions. Besides this, due to CL infections caused by different species in endemic areas, diagnostic methods enabling species identification are in great need. Species identification, in the time of diagnosis, is an important procedure for helping the clinicians in the planning of treatment as well as control measures. Polymerase chain reaction (PCR) is a specific and sensitive diagnostic tool that can also identify the parasite at species level. Kinetoplast DNA (kDNA) is one of the genetic regions that can be used for the detection of Leishmania parasites in clinical specimens, kDNA PCR is reported as one of the most sensitive methods related to species-specific variable regions in mini-circle long time ago. It has been considered as one of the most ideal targets for the diagnosis of leishmaniasis. The aim of the study was to perform PCR targeting kDNA by using the primers of Uni21/Lmj4 in clinical samples and compare the results with other parasitological methods like smear and culture, for the diagnosis of CL. The kDNA PCR, parasite culture and microscopical evaluation of stained smears of 62 specimens from suspected CL cases who have referred to Cutaneous Leishmaniasis Diagnosis and Treatment Center in Sanliurfa, Turkey were included in the study. The kDNA PCR showed the highest sensitivity 100% of the samples (35/35) among all diagnostic assays, followed by the microscopy (25/35 positive, 71.4% sensitivity) and culture (19/35 positive, 54.3% sensitivity). The sensitivity of combination of culture and microscopy was 88.6% (31/35 positive). These results suggested that performing kDNA PCR in addition to conventional techniques is important for improving the true diagnosis of CL to the species level and also important for establishing treatment regimens and designing appropriate precautions in highly endemic area like the southeastern region of Turkey.

[The importance of the contribution of rapid test, serological and molecular methods in the diagnosis of two imported malaria cases with atypical microscopy]. / Mikroskopide atipik görünümlü dis kaynakli iki sitma olgusunda hizli test, serolojik ve moleküler yöntemlerin taniya katkisinin önemi.

Malaria is a widespread and life-threatening disease in tropical and subtropical regions. In patients with typical clinical symptoms, malaria is considered as a preliminary diagnosis if there is a travel history to malaria-endemic areas. The basis of the laboratory diagnosis of malaria is the microscopic examination of Giemsa stained smears. On the other hand, the diagnosis and differentiation of Plasmodium species with microscopic examination may have some difficulties. In the first case, adifferent appearance from the classical Plasmodium vivax erythrocytic forms in infected erythrocytes were detected in 1% of all erythrocytes in thin smear blood preparations of a 26-year-old male with complaints of fever and chills and a story of travel to Nigeria. It was observed that parasitic nuclei were not prominent, and were located in the cytoplasm irregularly as chromatin or dye particles, nucleus fragments similar to Schüffner's granules in the form of scattered and granular spots were present in some erythrocytes, the cytoplasm of some Plasmodium erythrocytic forms were irregular and nuclei were not seen. There were no Schüffner's granules in any of the infected erythrocytes. P.vivax was detected by the rapid diagnostic test (OptiMAL, DiaMed GmbH, Switzerland), which searches for the antigens of Plasmodium species, in the peripheral blood sample of the patients. The P.vivax 18S rRNA gene was also detected by the multiplex real-time polymerase chain reaction. Antibodies against Plasmodium species were searched by using the Pan Malaria Antibody CELISA (CeLLabs Pty Ltd, Brookvale, Australia) kit in the patient's serum sample and the optical density (OD) value of the patient sample was measured five times the OD value of the positive control. In the second case, adifferent appearance from the classical P.falciparum erythrocytic forms in infected erythrocytes were detected in 12% of all erythrocytes in thin smear blood preparations of a 31-year-old male who has been suffering from persistent fever, severe headache, pain in the eyes and was known to be working in Nigeria. It was observed that some Plasmodium trophozoites have 1/3 of the size of erythrocytes such as P.vivax and have non-granular cytoplasm, some erythrocytic forms were round and the nucleus and cytoplasm were hardly distinguished, some of them were seen as crescent and close to the nucleus of the cytoplasm and some erythrocytic forms had characteristically a single nucleus and a scattered cytoplasm, similar to mature trophozoites of P.vivax. Although the Plasmodium young trophozoites were similar to P.vivax in means of magnitude, the forms in which the nuclei adhered to the erythrocyte wall were common. There were no P.falciparum gametocyte forms. P.falciparum like young trophozoite was observedonly in one of the four smears. P.falciparum was detected by the commercial rapid diagnostic test and P.falciparum 18S rRNA gene was also detected by the multiplex real-time polymerase chain reaction. Antibody formation against Plasmodium species was not detected in the ELISA test. In these case reports, the importance of the support of rapid diagnostic tests, serological and molecular methods to microscopic diagnosis and species determination of two imported malaria cases were demonstrated.

[The comparison of microscopy and real time polymerase chain reaction methods for the diagnosis of Pneumocystis Jirovecii pneumonia: evaluation of clinical parameters]. / Pneumocystis jirovecii pnömonisi tanisinda mikroskopi ve gerçek zamanli polimeraz zincir reaksiyonu yöntemlerinin karsilastirilmasi: Klinik bulgular ile yorumlanmasi.

INTRODUCTION: Pneumocystis jirovecii pneumonia (PCP) causes serious infections, especially in patients with immunosuppressive diseases. In this study, it was aimed to evaluate the results of samples obtained from PCP suspected patients using two different methods together with clinical data. MATERIALS AND METHODS: Microscopy and real time polymerase chain reaction (real time PCR) methods were performed with bronchoalveolar lavage (BAL) samples sended to Ege University Medical Faculty Direct Parasitology Diagnostic Laboratory between March 2009 and June 2010. Demographic characteristics, clinical and laboratory data were also recorded retrospectively. The data were evaluated using the SPSS 16.0 program. RESULT: A total of 42 BAL samples collected from patients (24 males, mean age: 31.49 ± 26.14) were included. There were totally 16 P. jirovecii positives either one of the tests. Sixteen and three samples were detected positive by real time PCR and microscopy, respectively. Trimethoprim-sulfamethoxazole was prescribed in 11 PCP diagnosed cases and 6 of them died. CONCLUSIONS: Today, despite the growing opportunities in diagnosis and treatment, PCP pneumonia is associated with high mortality. Careful examination of clinical data and immune status of the patients are important. Multidisciplinary approach is required for early PCP diagnosis.

Monitoring the Trends in Intestinal Parasite Frequencies; 2018 and 2022 Data

Objective: Monitoring intestinal parasite frequencies is effective on diagnosis, treatment, and prevention strategies to be developed against these parasites. In this study, it was aimed to reveal the parasite species and frequency data of stool samples in parasitology direct diagnosis laboratory. Methods: Stool parasitological examination results were obtained retrospectively from our laboratory internal quality control data tables. Data belonging to the year 2018 and 2022 were compared retrospectively. Results: Annual parasites detected in stool samples were 388 of 4.518, and 710 of 3.537, in 2018 and 2022, respectively. Frequency of parasite detection in stool samples was found to be significantly higher in 2022 (p<0.0001). Number of stools with more than one parasite was 12 and 30 in 2018 and 2022, respectively. Incidence of infection with more than one parasite was significantly higher in 2022 (p=0.0003). Five most common parasite species were Blastocystis spp., Enterobius vermicularis, Cryptosporidium spp., Giardia intestinalis and Entamoeba histolytica in 2018, respectively; and Cryptosporidium spp., Blastocystis spp., Cyclospora spp., Entamoeba dispar and Giardia intestinalis, in 2022, respectively. Cryptosporidium spp., Cyclospora spp. and Entamoeba dispar increased significantly, while Blastocystis spp. and Enterobius vermicularis decreased significantly, in 2022. Conclusion: According to the data obtained, causative agents for intestinal parasitic infections were protozoans, especially Cryptosporidium spp. It has been concluded that tightening the measures for protection of water with one health approach and improving the education and habits of society on personal hygiene and food safety can be effective in reducing the frequency of intestinal parasite infections in our region.

Unpleasant Souvenir: Imported Plasmodium falciparum Malaria in Türkiye.

Objective: Each year, approximately 125 million people visit malaria-endemic countries. This study aimed to investigate the clinical characteristics of imported Plasmodium falciparum malaria infections in Türkiye. Methods: The study included patients diagnosed with P. falciparum malaria between 1996 and 2022. A retrospective evaluation was conducted on whole blood samples and/or blood smears, as well as detailed medical histories, clinical manifestations, and laboratory findings. A total of 131 imported cases of P. falciparum were included in the study. Results: Among the patients, 121 were male. Of these, 101 had traveled to Africa, while 30 had visited Asia. Among the patients, 109 were returned travelers, and 22 were refugees/migrants. Early trophozoites were observed in all patients, while gametocytes were detected in 30 patients. Cerebral malaria developed in 15 patients, resulting in the death of two individuals. Additionally, 10 patients received preventive chemoprophylaxis. Conclusion: Turkey is situated on migration routes that connect two continents to Europe, where more than 95% of the global malaria burden exists. The importation of malaria through returned travelers poses a risk of malaria reintroduction in our country, given the presence of suitable vectors, climate conditions, and environmental factors. Importantly, 30 patients (22.9%) exhibited gametocyte forms of P. falciparum, which have the potential to infect Anopheles species, thus establishing a basis for local malaria transmission.

Investigation of in vitro Efficacy of Miltefosine on Chronic Cutaneous Leishmaniasis

Objective: Leishmaniasis is the second deadliest parasitic disease in the World Health Organisation's list of neglected diseases, following malaria. Cutaneous leishmaniasis (CL) is the most common form of the disease and it is one of the few communicable diseases with increasing incidence rates owing to factors like armed conflicts and climate change. CL can be divided into two major groups: Acute CL (ACL) and chronic CL (CCL). The aim of this study was to compare the in vitro efficacy of miltefosine and pentavalent antimony compounds in the CCL patient samples. Methods: Five isolates previously isolated from 5 CCL patients were included in this study. Genotyping is performed using internal transcribed spacer 1 (ITS 1) gene region real-time PCR. In vitro drug efficacy tests were applied to determine their activity against meglumine antimoniate (MA) and miltefosine. Serial dilutions (512, 256, 128, 64, 32, 16, 8 and 4 µg/mL) prepared from MA and miltefosine were prepared in 96-well flat-bottom cell culture plates and incubated at 24 °C for 48 hours. The efficacy of the drug on Leishmania spp. promastigotes after 24 and 48 hours was evaluated by hemocytometer slide and XTT cell viability test. Results: All of the samples were genotyped as L. tropica. Evaluation of 24 and 48 hours showed, 128 µg/mL and 256 µg/mL and 32 µg/mL and 64 µg/mL concentrations of miltefosine and MA were enough to kill all the promastigotes respectively. The results of the hemocytometer slide and XTT were consistent. Conclusion: There are no studies investigating the in vitro efficacy of miltefosine with the CCL patient group. To overcome the treatment challenges experienced in this special patient group, more studies are needed. According to our results, it is concluded that miltefosine is efficient for the treatment of CCL and further clinical studies with miltefosine will reveal valuable data.

Autochthonous transmission of Leishmania donovani and Leishmania major with all the components of infection cycle at Europe's doorstep.

OBJECTIVES: Leishmaniasis is a vector-borne disease and dogs may act as urban reservoirs. Turkey and most of the Mediterranean basin countries are endemic for leishmaniasis. In this study, it is aimed to report the autochthonous leishmaniasis cases, with all the components of the infection cycle (reservoir, vector, and the host) in a region close to Europe. METHODS: Nine human and four canine autochthonous leishmaniasis cases were included in the study. Direct microscopy, culture methods, serological, and molecular tests were applied to the samples obtained from the cases. RESULTS: VL and CL patients consisted of 2 L.infantum, 1 L. donovani, 2 L. tropica, and 2 L. tropica,1 L. major,1 L. infantum infected patients respectively. CanL cases were infected with L. infantum, L. donovani, L. tropica, and L. major. CONCLUSIONS: All the cases were autochthonous cases located in Manisa province. As Greece and all the Mediterranean basin countries in Europe share competent vectors, it is concluded that the detection of all 4 species of Leishmania parasites in such proximity to Europe poses an important public health threat for Europe. This study reports all four species of Leishmania spp., including L. major and L.donovani in close proximity to continental Europe.

Biosafety Risk Assessment of a Routine Diagnostic Laboratory During the Coronavirus Disease 2019 Pandemic.

Background: In this study, we aimed to perform a biosafety risk assessment to determine measures to be taken against coronavirus disease 2019 (COVID-19) in the routine diagnostic parasitology laboratory of a tertiary health care center. Methods: The risk assessment template included in the supplement of the interim guidance of "WHO Laboratory Biosafety Guidance Related to COVID-19" was used for the risk assessment. Risk assessments were carried out for the "diagnosis of protozoan diseases in respiratory tract samples" and "diagnosis of intestinal parasitic diseases" processes. Initial risk of the laboratory activities was determined before additional risk control measures and overall initial risk was estimated for each process. Overall residual risk of the laboratory activities after risk control measures was estimated for each process. Results: Overall initial risk for both processes was "very high." Fresh microscopic examination steps in both processes and concentration steps for "diagnosis of intestinal parasite diseases" were discontinued. All aerosol-generating steps were moved into a class-IIA biological safety cabinet. Overall residual risk was "medium" for both processes. Conclusion: This study serves as an example for clinical laboratories regarding how the risk assessment approach in guidelines can be transferred to daily practice.

Evaluating the Glucantime Concentration for the ex vivo Glial Cell Model of Antimony-resistant Leishmania tropica Amastigotes

Objective: Because the protocols used in the treatment of leishmaniasis can be toxic and have many limitations, such as the development of resistance against such protocols, new treatment options are needed, especially against resistant patients. Ex vivo models may be a good source for evaluating new drug options for patients with antimony-resistant parasites. This study aimed to evaluate the Glucantime concentration for our ex vivo glial cell amastigote model we had defined in previous work. Methods: We prepared the astroglial cell culture from brains of 2 to 3 day old neonatal Sprague-Dawley rats under sterile conditions by modifying McCarthy's method. Four plates of cells were infected with antimony-resistant Leishmania tropica promastigotes. After 24 h of incubation, we added Glucantime to 3 plates with different concentrations. After 72 h, we removed the supernatant and then dried, fixed, and stained the plates with Giemsa to count the amastigotes in the glial cells. Results: We observed the amastigotes in glial cells in the control flask. Glial cells were ruined in flasks, which include 75 µg/mL and 37.5 µg/mL Glucantime. The number of amastigotes per 100 glial cells was 116 for the flask with 7.5 µg/mL Glucantime concentration, while 487 for the control flask. Conclusion: We found that while high concentrations of Glucantime were toxic for glial cells, 7.5 µg/mL Glucantime concentration managed to reduce the number of Leishmania tropica amastigotes in glial cells.

Overcoming the Challenge; In Vivo Efficacy of Miltefosine for Chronic Cutaneous Leishmaniasis.

BACKGROUND: Cutaneous Leishmaniasis (CL) is the most common form of leishmaniasis. CL can be divided into two major groups: acute CL (ACL) and chronic CL (CCL). The aim of this study is to compare the efficacy of miltefosin and pentavalent antimony compounds in vivo with the CCL patient samples. MATERIALS: Three study groups were formed, each consisting of five male Mus musculus (Balb/C) mice. In this model, promastigotes from the culture of a CCL patient were utilized. 100 µL L. tropica promastigote suspension with a density of 108 promastigotes/ml were injected into the hint-right footpad of each experimental animal intradermally. Footpads of the mice were measured every two weeks until 24th week. From the 13th week, miltefosin 50 mg/kg/day was administered orally using gavage for 21 days, Meglumin antimoniate (MA) was administered by intramuscular (IM) injection daily for 21 days at 50 mg/kg/day and saline was administered IM for 21 days for the miltefosine, MA and control group, respectively. RESULTS: The footpad measurements of the miltefosine group were lower than the control group statistically. Between the MA group and the miltefosine group and MA group and the control group, there was no statistically significant difference. Giemsa stained slides revealed amastigotes in one, two and all of the slides for the miltefosine, MA and control group, respectively. Molecular tests were performed with the Rotor-Gene device and L. tropica consistent peaks were obtained in one of the miltefosine group, four in the MA group and all mice in the control group. CONCLUSIONS: Demonstration of both clinical and laboratory improvement in four of the five experimental animals provides strong evidence that miltefosine is an effective drug in the treatment of CCL. In the literature, no clinical or laboratory studies using miltefosine have been performed with CCL patients only.

Comparison of Skin Scraping and Standard Superficial Skin Biopsy in the Laboratory Diagnosis of Scabies

Objective: Scabies is diagnosed based on the presence of burrows on the skin, Sarcoptes scabiei adult, egg, or scybala in skin scrapings. The laboratory diagnosis of scabies poses various challenges. We aimed to compare the analytical performance of skin scraping and standard superficial skin biopsy (SSSB) and to investigate the correlation with false negative results in the laboratory diagnosis of scabies. Methods: Skin scraping and SSSB were applied from July 1 to December 31, 2018 on 42 patients whose burrows were marked using dermatoscopy, as obtained from the laboratory information system. Results: The number of patients who tested positive for scabies with skin scraping was 18 (42.9%) and 24 (57.1%) with SSSB, and the difference was significant (p=0.003). Sensitivity was 42.9% for skin scraping and 57.1% for SSSB. The number of positive cases with both techniques was 15 (35.7%). The number of patients positive with only skin scraping was 3 (7.1%) and only SSSB was 9 (21.4%). Conclusion: To date, it has seemed impossible to diagnose scabies using a single clinical or laboratory test. According to our results, SSSB is an inexpensive and easy-to-apply method with high sensitivity for obtaining skin samples for scabies laboratory diagnosis.

Intestinal Protozoan Parasitic Infections in Immunocompromised Child Patients with Diarrhea.

Intestinal protozoan parasites are common causes of infectious diarrhea in children receiving anticancer therapy or undergoing transplantation. Additionally, immunosuppression therapy in such patients may exacerbate the symptoms related to these parasitic infections. The aim of this study was to evaluate the prevalence and diagnostic importance of parasitic protozoan infections in children treated for malignancies or undergoing transplantation, and to highlight the control of intestinal parasitic infections for immunosuppressed patients at a hospital in Izmir, Turkey. In total, 82 stool samples from 62 patients were analyzed by microscopic examination and polymerase chain reaction (PCR) for the presence of coccidian parasites. Our results showed that Cryptosporidium, Cyclospora, and Cystoisospora were present in 22.5% (14/62), 9.6% (6/62), and 3.2% (2/62) of the cases using either method, respectively. The prevalence of these coccidian parasites identified with both methods was 35.4% (20/62). Other intestinal parasites (Blastocystis, Giardia, and Entamoeba coli) were detected in 10 patients. PCR analysis showed the presence of all coccidian parasites in the same stool sample for one patient. Finally, both PCR and microscopic examination of the stools revealed that there is a higher prevalence of Cryptosporidium, Cyclospora, and Cystoisospora in immunocompromised children. These examinations allowed an early start of appropriate antibiotic treatments and led to an increased percentage of correctly treated patients.

Lophomonas blattarum Associated Broncho-Pulmonary Infection After Immunotherapy: A Case Report and A Smart-phone Based Video of Trophozoite

The frequency of bronchopulmonary protozoan infections has raised due to increased number of immunosuppressed patients in recent years. One of them is Lophomonas blattarum which is a multi-flagellated protozoan parasite of termites and several cockroach species. The drug regimens commonly used in bronchopulmonary infections are not effective against L. blattarum. Therefore, rapid and accurate diagnosis of L. blattarum infection is of great importance in the treatment success. The laboratory diagnosis of L. blattarum infection is made on the basis of observation of the characteristic trophozoite in various samples. It is of a great importance to distinguish the protozoon from ciliated respiratory epithelium to avoid wrong positivity. The presented case developed an acute respiratory distress syndrome a short while after taking nivolumab immunotherapy. The morphological features of L. blattarum were demonstrated by examining the bronchoalveolar lavage fluid of the patient under light microscopy. Additionally, URL (https://youtu.be/EQIAsFl6AJY) of a smart-phone based video of trophozoite of this patient was added into this report.

The Prevalance of Blastocystosis among Patients with Gastrointestinal and Dermatologic Complaints and Effects of Blastocystis spp. Density on Symptomatology.

OBJECTIVE: Blastocystosis has been linked with non-specific symptoms, such as diarrhea, abdominal pain, and distention. In this study, we evaluated the relationship between Blastocystis spp. with urticaria and intestinal symptoms. METHODS: The results of the stool examinations of the patients who were referred to Ege University Medical Faculty Hospital's Medical Parasitology Department Direct Diagnosis Laboratory with gastrointestinal (GIS) and/or dermatologic symptoms between January 2011 and July 2016 were retrospectively scanned. RESULTS: Of the evaluated 37108 stool samples, 2573 (6.93 %) were identified to be positive for Blastocystis spp. The patients with gastrointestinal complaints comprised 68.4% of Blastocystis spp. positive samples (1.761 samples) while 30.1% of patients had dermatologic symptoms (urticaria) (776 samples). Blastocystis spp. density in the non-amplified (without using any stool concentration technique) stool samples of the patients with GIS and dermatological symptoms was as follows: 2.47%, 1.35% rare, 21.73%, 22.17% few, 49.65%, 54.29% medium, 26.27%, and 22.17% dense, respectively. CONCLUSION: 75.92% and 76.46% of the patients with GIS and dermatological complaints had medium to dense parasite densities in their stool samples respectively. This suggests a positive correlation between parasite density and GIS and dermatologic symptomatology.

Intestinal Protozoa in Hiv-Infected Patients: A Retrospective Analysis.

OBJECTIVE: Intestinal parasitic infections are common in immunodeficient patients. In developing countries, the incidence of diarrhea due to parasitic infections in HIV (human immunodeficiency virus)-positive individuals is reported to be over 90%. The present study aimed to investigate the presence of intestinal protozoa in HIV-positive patients with gastrointestinal complaints. METHODS: The fecal samples of 65 HIV-positive patients (14 women, 51 men) were included. Clinical data obtained from patients' files and laboratory results were retrospectively scanned using laboratory information system. Age, sex, parasite positivity, CD4+ count, HIV RNA level, and antiretroviral therapy information were recorded. RESULTS: Fourteen Cryptosporidium spp. (21.5%), 2 Cyclospora spp. (3.1%), 7 Blastocystis spp. (10.8%), and 1 Cryptosporidium spp.+Blastocystis spp. (1.5%) were detected. The median duration of antiretroviral treatment was 3 months and 12 months in patients with and without parasites in fecal samples, respectively. The duration of antiretroviral treatment was significantly higher in non-infected patients (p=0.002). No significant correlations were found between parasite presence and CD4+ T cell counts or HIV RNA levels. CONCLUSION: Our findings suggest that positive effects of antiretroviral therapy on the immune system of HIV-infected patients reduce the risk of intestinal parasitic infection, and thus, this treatment may play an important role in protection.

Human myiasis in patients with diabetic foot: 18 cases.

BACKGROUND: Myiasis complication of diabetic foot ulcer has only been presented in a few case reports. Therefore, there is a need for additional data on this infestation. OBJECTIVE: Evaluate clinical characteristics of human myiasis in patients with diabetic foot. DESIGN: Case series. SETTINGS: A tertiary referral healthcare institution and a diabetic foot center. PATIENTS AND METHODS: Patients with diabetic foot infection com.plicated by myiasis who were admitted between June 2012 and July 2017. MAIN OUTCOME MEASURES: Bacterial infection rate, accompanying bacterial agents, amputation (morbidity) and mortality rate. SAMPLE SIZE: 18. RESULTS: Eight (44.4%) of the patients were female. Sixteen (88.9%) had moderate-to-severe infections; 15 (83.3%) had necrotic tissue. Larval debridement therapy was performed on all patients at the bed.side in consecutive sessions. A third-stage larva of Calliphora was detected in one case (5.6%). Second- and third-stage larvae of Lucilia sericata were detected in 5 (27.8%) and 7 (38.9%) patients, respectively. All the patients had a bacterial infection with myiasis. Twelve (66.7%) patients underwent amputation. Three (16.7%) patients died. Myiasis was more frequent in the months of May, June and July. CONCLUSION: To our knowledge, this is the largest reported series of cases of diabetic foot with myiasis. The most common parasitic agent was Lucilia sericata. Bacterial soft tissue infections were observed in all cases. Poor hygienic conditions were noteworthy and all patients were in need of radical surgery. Myiasis complication of diabetic foot is more frequently seen in the spring and summer. LIMITATIONS: Insufficient follow-up time for analysis of possible confounding factors. CONFLICT OF INTEREST: None.

The association between Cytomegalovirus co-infection with Pneumocystis pneumonia and mortality in immunocompromised non-HIV patients.

INTRODUCTION: Impact of Cytomegalovirus (CMV) co-infection pneumonia in non-HIV patients with Pneumocystis jirovecii pneumonia (PCP) is unclear. OBJECTIVES: The aim of our study was to determine whether CMV co-infection is associated with an increased risk of mortality. METHODS: Our study was conducted at Ege University Hospital, Turkey. We used molecular assays to diagnose Pneumocystis jirovecii in respiratory samples, and CMV in both respiratory and blood samples. We compared morbidity and mortality stratified by CMV co-infection status. RESULTS: Between 2009 and 2015, 43 patients (mean age: 56.7 ± 15.3 years) were diagnosed with PCP. Only 3 of 43 patients had received PCP prophylaxis. We microbiologically confirmed CMV co-infection in 28 of 43 (65.1%) patients. Acute respiratory distress syndrome (ARDS) and requirement of mechanical ventilation were more common in the CMV co-infection group (P = .019 and P = .031 respectively), and duration of intensive care unit was also longer (P = .006). In univariate analyses, mortality at 30 days was higher in the CMV co-infection group as compared to the group with PCP alone (78.6% and 46.7% respectively; P = .046). In multivariate analyses, mortality was independently associated only with the presence of ARDS [OR: 6.22 95% CI 1.3-29.32] and the association with CMV co-infection was no longer significant [OR: 2.6 95% CI 0.49-13.72, P = .257]. CONCLUSION: The risk of mortality appears to be increased in the setting of CMV and PCP co-infection in HIV-uninfected immunocompromised patients. PCP prophylaxis use was lower than expected, suggesting low physician awareness of the risks of PCP in this population.

The Concept of Travel Medicine and the Actual Situation of Travel-Related Illnesses.

Travel medicine defines all diseases and medical situations that are related to travel. Travel medicine comprises infectious diseases, traumas, altitude sickness, sun burns, embolisms, jet lag, and many more travel-related situations. With the increasing possibility and ease of travel, the number of people who have travelled internationally has exceeded 1.13 billion in 2014, and the revenues of international travel have exceeded 1.25 trillion dollars. With every passing day, international travels are shifting toward the developing countries and to more exotic regions of the world, and travelers tend to be more adventurous and daring, thereby increasing risky behaviors during travels. Traveling plays an important role in transmitting infections such as Zika virus infection, Ebola, avian flu, severe acute respiratory syndrome, Chikungunya, and dengue fever and is the principal reason for the epidemics of these types of infections on a global scale. With this background, we suggest that travel medicine is an important but "neglected" medical discipline as the discipline of Parasitology itself like most parasitic diseases.