Pneumocystis jirovecii Pneumonia in Solid-Organ Transplant Recipients: A National Center Experience.

OBJECTIVES: Immunosuppressive therapies have impro-ved survival in solid-organ transplant recipients at the expense of increased prevalence of opportunistic infections. We investigated the prevalence, risk factors, and prognosis of Pneumocystis jirovecii pneumonia in solid-organ transplant recipients who were followed by our transplant unit. MATERIALS AND METHODS: We conducted a retrospective observational study using medical record reviews to identify all adult solid-organ transplant recipients who underwent bronchoscopy and bronchoalveolar lavage between January 2011 and 2018. We collected clinical characteristics, including risk factors and prognosis. Pneumocystis jirovecii pneumonia symptoms com-patible with microscopy and/or positive nucleic acid amplification assays were defined as proven infection by P. jirovecii pneumonia. RESULTS: We identified 312 adult solid-organ transplants (114 renal, 1 kidney and pancreas, 197 liver) in this period. Overall, 113 (36.2%) pulmonary disease consultations were performed in the posttransplant stage, and 46 (40.7%) patients underwent bronchoalveolar lavage with P. jirovecii screening. We identified 18 patients who tested positive for P. jirovecii infection according to nucleic acid amplification assay; 3 were not proven, and 7 had a transplant date before 2011. The prevalence was 8/312 (2.6%); of these 8 patients, 5 had the same genotype cluster. Median P. jirovecii pneumonia development time was longer in renal transplant recipients (P = .016). Only renal transplant recipients were offered Pneumocystis prophylaxis for 6 months. Concomitant viral infection including cytomegalovirus was the only significant factor for P. jirovecii pneumonia development (P = .028). Intensive care admission was 40% (n = 6), and disease-related mortality was 33% (n = 5). CONCLUSIONS: The overall prevalence of P. jirovecii pneumonia in solid-organ transplant recipients was similar to other single-center reports. Prophylaxis prevented early posttransplant P. jirovecii pneumonia. However, P. jirovecii pneumonia may develop at any posttransplant stage, and viral infections other than cytomegalovirus should also be considered as a predictor.

Absence of dihydropteroate synthase gene mutations in Pneumocystis jirovecii strains isolated from Aegean region of Turkey.

Sulfonamide group drugs and their antimetabolite combinations are the most preferred drugs in the treatment and prophylaxis of Pneumocystis jirovecii pneumonia (PCP). Especially with the long-term use of trimethoprim-sulfamethoxazole (TMP-SMX) and dapsone, certain point mutations in the Pneumocystis jirovecii (P. jirovecii) dihydropteroate synthase (DHPS) gene are known to play an important role in the development of resistance. In the present study, we investigated the 165th and 171st nucleotide mutations in the DHPS gene in the P. jirovecii isolates from immunosuppressed and immunocompetent cases. P. jirovecii isolates from the bronchoalveolar lavage fluid (BALF) samples of 31 hospitalized cases and lung tissue samples of 37 autopsy cases were included in the study. For the analysis of wild-type and mutant genotypes, after the touchdown-PCR amplification method, the restriction fragment length polymorphism (RFLP) method was used. In this study, P. jirovecii DHPS gene was amplified in 28 of 68 (41%) of the samples. The RFLP method revealed that all the isolates in which the DHPS gene was amplified were considered as wild-type genotypes. To our knowledge, this present study is the first study in Turkey investigating P. jirovecii DHPS gene mutations associated with the sulfonamide resistance. All the isolates showed a wild-type pattern indicating that the occurrence of P. jirovecii DHPS mutations in Turkey is very low or absent.

Investigation of parasitic and viral pathogens in mussels (Mytilus galloprovincialis) in the Gulf of Izmir, Turkey.

OBJECTIVES: To investigate Microsporidia spp. parasite, hepatitis A virus (HAV), and norovirus (NoV) contamination in mussels collected from 8 stations in the inner, middle, and outer regions of the Gulf of Izmir. METHODS: In this cross-sectional study carried out between August 2009 and September 2010 in the Gulf of Izmir, Turkey, 15 mussels collected from each of the stations each season were pooled and homogenized to create a single representative sample. Thirty representative samples were available for analysis. Direct polymerase chain reaction (PCR), RT-nested PCR, and RT-booster PCR were used to investigate the pathogens.  RESULTS: The mussels were negative for Microsporidia spp., but 8 (26.7%) samples analyzed were positive for HAV and 9 (30%) were positive for NoV. Excluding Foca and Gediz, viral contamination was detected in all of the stations sampled.  CONCLUSION: Our results suggest that viral contamination is present in mussels in the Gulf of Izmir and may pose a potential threat to human health in the region. Necessary measures should be taken to prevent future illness due to these pathogens.

[Investigation of the in vitro effects of Melissa officinalis L., Mentha x piperita L. and Ocimum basilicum L. (Lamiaceae) essential oils on the cysts and trophozoites of Acanthamoeba castellani]. / Melissa officinalis L., Mentha x piperita L. ve Ocimum basilicum L. (Lamiaceae) uçucu yaglarinin Acanthamoeba castellani kist ve trofozoitleri üzerine in vitro etkisinin arastirilmasi.

Acanthamoeba species are free living amoeba found widely all over the world. They are responsible for Acanthamoeba keratitis (AK), an infection which is especially seen in contact lens users and after minor corneal traumas, that may lead blindness. At present, antifungals and antiseptics are used for the treatment of AK cases, however, some problems such as long treatment periods and the occurrence of side effects, resistance of cyst forms against drugs, emphasize the need for new drugs. There are some published studies that pointed out the effectiveness of plant extracts and essential oils on Acanthamoeba spp. The aim of this study was to investigate the in vitro effects of essential oils of Mentha x piperita L. (peppermint), Melissa officinalis L. (lemon balm) and Ocimum basilicum L. (basil) belonging to Lamiaceae family, on the cysts and trophozoites of Acanthamoeba castellanii. The strain used in our study, namely A. castellanii T4 genotype, is the most frequently isolated amoeba from environment and also the causative agent of AK and granulomatous amebic encephalitis. For the determination of amebicidal activity, essential oils obtained from Mentha x priperita L., Melissa officinalis L. and Ocimum basilicum L. by Neo-Clevenger type of distillation apparatus have been used. In vitro experiments were performed by using 96-well microplates. Cyst and trophozoite solutions were added on the essential oil dilutions to obtain the last concentrations of 40, 20, 10, 5, 2.5 and 1.25 µg/ml for the cysts, and 10, 5, 2.5, 1.25, 0.625 and 0.313 µg/ml for the trophozoites. After the incubation of microplates at 30oC for 1, 6, 24, 48 and 72 hours, the viability of parasitic forms were evaluated under the light microscope followed by staining trypan blue. It was found that, each essential oil showed amebicidal effect on A.castellani cysts and trophozoites dependent on dosage and time, when compared with the control group, The maximum lethal effect occured with Melissa officinalis followed by Mentha x piperita and Ocimum basilicum, respectively. In our study, susceptibility of A.castellanii trophozoites to essential oils were more than the cysts, as expected. The essential oils of Melissa officinalis and Mentha x piperita showed 100% lethal effect at their highest concentrations whereas the essential oil of Ocimum basilicum showed only 63.3% lethal effect on cysts after 72 hours at the highest concentration (40 µg/mL). The results of this first study investigating the activities of essential oils extracted from Mentha x piperita, Melissa officinalis and Ocimum basilicum against Acanthamoeba spp. cysts and trophozoites, have suggested that, these essential oils could be potential novel and alternative natural products for the treatment of Acanthamoeba spp. infections.

[Pneumocystis jirovecii Pneumonia in a Patient with Interstitial Lung Disease]. / Interstisyel Akciger Hastaligi Zemininde Gelisen Pneumocystis jirovecii Pnömonisi.

Pneumocystis pneumonia (PCP) caused by Pneumocystis jirovecii (P. jirovecii) is an opportunistic pulmonary infection that occurs in immunocompromised patients. Here, a 49-year-old female patient who was admitted to our hospital with respiratuary distress and whose bronchoalveolar lavage (BAL) fluid specimens had P. jirovecii and Aspegillus fumigatus was presented. She had been treated with corticosteroids because of interstisial lung disease and she was also diabetic. It is important to define the coinfection developed in the presence of immunosuppression.

[Investigation of Pneumocystis jirovecii colonization in patients with pulmonary diseases]. / Akciger Hastaligi Olan Hastalarda Pneumocystis Jirovecii Kolonizasyonunun Arastirilmasi.

OBJECTIVE: The detection of Pneumocystis jirovecii or its DNA in respiratory samples from individuals who do not have signs or symptoms of pneumonia has been defined as colonization. In this study, we aimed to investigate the prevalence of P. jirovecii colonization in patients with various lung diseases. METHODS: Thirty patients who were followed-up and who had undergone bronchoscopy for diagnosis of different underlying diseases or pulmonary signs were included in the study. Bronchoalveolar lavage (BAL) fluids of these patients were analyzed with nPCR amplification of the mt-LSUrRNA gene of P. jirovecii. In addition to nPCR, giemsa, Gomori's methenamine silver (GMS), and indirect fluorescence antibody (IFA) staining assays were applied to all samples. RESULTS: P. jirovecii DNA was detected in 21 of 30 (70%) BAL samples by nPCR. However, P. jirovecii cysts were found in 1 of 21 nPCR-positive samples by giemsa and GMS. IFA assay showed six samples to be positive, but only four of them were found to be positive by nPCR. CONCLUSION: Result of our study showed that prevalence of P. jirovecii colonization is particularly high in patients with chronic pulmonary diseases, and nPCR was a good assay for evaluation of the colonization of P. jirovecii.