Investigating the Antifungal Effect of the Essential Oil of Thymus eriocalyx on Dominant Filamentous Fungal Agents Isolated from Livestock and Poultry Feed.

BACKGROUND: One of the most important principles in disease control is the health of livestock and poultry feed. Given the natural growth of Th. eriocalyx in Lorestan province, its essential oil can be added to the livestock and poultry feed and prevent the growth of the dominant filamentous fungi. OBJECTIVE: Therefore, this study aimed to identify the dominant moldy fungal agents of livestock and poultry feed, examine phytochemical compounds and analyze antifungal effects, anti-oxidant properties, as well as cytotoxicity against human white blood cells in Th. eriocalyx. METHODS: Sixty samples were collected in 2016. The PCR test was used to amplify ITS1 and ASP1 regions. The analysis of essential oil was conducted by gas chromatography and gas chromatographymass spectrometry devices. MIC and MFC were performed using the broth micro-dilution method. For the analysis of DDPH activity, DDPH was used. Cytotoxicity effect on healthy human lymphocytes was carried out by the MTT method. RESULTS: In this study, A. niger, F. verticilloides and F. circinatum, P. oxalicum, and P. chrysogenum were the most resistant species, and A. oryzae and A. fumigatus, F. prolifratum and F. eqiseti, P. janthnellum were the most susceptible ones. IC50 value of T. daenensis Celak was 41.33 µg/ml, and 100 µl/ml of the essential oil caused slight cell lysis. CONCLUSION: Considering our results, compared with drugs and chemical additives, essential oils can be added to livestock and poultry feed to prevent the growth of filamentous fungi in the livestock and poultry feed.

Changes in the expression of miR-103a and miR-21: a functional diagnosis of toxocariasis in rats.

Introduction. Toxocariasis is a zoonotic parasitic disease caused by migrating nematode worms, Toxocara species larvae, within tissues. MicroRNAs (miRNAs) are small RNA molecules that regulate gene expression at a post-transcriptional level.Hypothesis/Gap Statement. miRNA-based diagnostic biomarkers for toxocariasis are emerging, but there is limited information about the role of many miRNAs and a more detailed diagnostic evaluation of miRNA expression patterns is needed to understand their immunobiological function.Aim. We investigated the expression levels of circulating miRNA 21 and miRNA 103a as potential biomarkers for the prediction and diagnosis of toxocariasis in Wistar rats infected with Toxocara canis.Methodology. Thirty Wistar rats were inoculated orally with 2500 T. canis embryonated eggs via gavage. Serum samples were collected from infected animals and were tested against T. canis antigens for 60 days post-infection. The plasma samples were isolated for quantitative real-time PCR (qPCR) assays and qPCR was used to assess transcription levels of miRNA 21 and miRNA 103a.Results. The prevalence of anti-Toxocara IgG was detected in 7/30 (23.3 %) infected rats. Molecular analysis of miRNAs 21 and 103a showed that expression levels of miRNAs in both groups of Toxocara-positive and negative samples were the same without significant association. The ratio of housekeeping gene expression (U6) to gene expression of miRNAs 21 and 103a indicated the rate of change (1/1.38 ≈ 0.75 and 1/0.751 ≈ 1.3, respectively).Conclusion. Our study revealed that miRNAs 21 and 103a might play fundamental roles as biomarkers and diagnostic tools for toxocariasis. However, the changes in expression of these miRNAs were not adequate to be used as biomarkers in diagnosis.

Clinical impact of Candida respiratory tract colonization and acute lung infections in critically ill patients with COVID-19 pneumonia.

Coronavirus disease 2019 (COVID-19), which is attributable to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has been causing a worldwide health issue. Airways colonization by Candida spp. is prevalent among patients on automatic ventilation in intensive care units (ICUs). This research aimed to ascertain the risk factors and roles of Candida spp. respiratory tract colonization, and Candida lung infection during the progression of COVID-19 pneumonia in critically ill patients. In total, Candida spp. were recovered in 69 from 100 immunosuppressed patients with COVID-19. Bronchoscopy was used to collect the Bronchoalveolar lavage (BAL) specimens. For the identification of Candida spp. PCR sequencing was done using the ITS1 and ITS4 primers. The amplification of the HWP1 gene was conducted to identify the Candida albicans complex. The antifungal activities of fluconazole, itraconazole, voriconazole, amphotericin B and caspofungin against Candida spp. were evaluated using the Clinical and Laboratory Standards Institute M60. In 63.77% of the patients, Candida respiratory colonization at D0 and D14 had no impact on the severity of COVID-19. In comparison to C. albicans strains, Candida respiratory disorder with C. glabrata had influenced the severity of COVID-19 for critically ill patients following adjustment for the risk factors of COVID-19 (P < 0.05). Amphotericin B and caspofungin showed superior activity against all Candida spp. All antifungal agents showed 100% sensitivity against the two C. africana strains. Our observation on patients who used automatic ventilation, respiratory colonization by Candida spp. was not seen to influence the infection or death caused by COVID-19. Amphotericin B and caspofungin showed superior activity against all Candida spp. and were recommended for the treatment regime of pulmonary candidiasis associated with COVID-19 infection. Although "Candida pneumonia" is rarely being reported in critically ill patients, Candida airway colonization mainly by Candida albicans is common especially among patients with diabetes, malignancies, and kidney disorders.

Allergic fungal rhinosinusitis caused by Neoscytalidium dimidiatum: A case report: Allergic fungal rhinosinusitis due to Neoscytalidium dimidiatum.

Neoscytalidium dimidiatum is a rare dematiaceous fungus that was first described in 1916 as Dothiorella mangiferae. From the standpoint of epidemiology and therapy, early detection of fungal rhinosinusitis (FRS), the causative agents, and their associated risk factors can improve the therapeutic outcome and decrease the mortality rates among patients. In this study, we report a 34-year-old Iranian female patient with allergic bronchopulmonary aspergillosis (ABPA), who presented to our facility with an 8-year history of chronic fungal sinusitis, drug-resistant asthma, pneumonia, bronchitis, post-nasal discharge, nasal obstruction, nasal polyposis, and anemia. The patient was subjected to diagnostic nasal endoscopy and computed tomography (CT) scan of paranasal sinuses, as well as routine, complementary mycological, and molecular methods, which confirmed the diagnosis of allergic fungal rhinosinusitis in patients with ABPA. Neoscytalidium dimidiatum was isolated from the sinus of the patient. Results of in vitro susceptibility tests indicated that the case isolate was susceptible to amphotericin B and itraconazole at concentrations which are commonly achieved in patients receiving recommended dosages for invasive mycoses (0.25 to 0.75 mg/kg of body weight daily for amphotericin B and 100 to 400 mg daily for itraconazole) and resistant in vitro to caspofungin, voriconazole, and posaconazole. The patient was successfully treated with amphotericin B / itraconazole + postoperative oral corticosteroids (OCS). Neoscytalidium dimidiatum infection should be considered as a possible additional factor in the etiology of AFRS, especially in immunocompromised patients.

Acute invasive fungal rhinosinusitis: Molecular identification and update in management of frozen section biopsy.

The clinical diagnosis of Acute Invasive Fungal Rhinosinusitis (AIFRS) is technically difficult because it presents with non-exclusive and nonspecific clinical symptoms. Laboratory confirmation (usually via histopathologic techniques such as formalin-fixed paraffin-embedded (FFPE)) is necessary but it is time-consuming, despite the urgent need for timely diagnosis of AIFRS for effective management. This study aimed to investigate the sensitivity and specificity of the GMS frozen-section biopsy in the diagnosis of AIFRS and compare the same with that of different tissue staining methods to provide valid decision-grounds that may guide clinicians in prompt diagnosis of acute fungal invasive rhinosinusitis. A cross-sectional study was conducted in the Medical Mycology Laboratory, Faculty of Medicine, Iran University of Medical Sciences between 2018 and 2020 on 200 patients with suspected AIFRS referred to Baqiyatallah and Imam Khomeini Hospital, Tehran. All patients were subjected to diagnostic nasal endoscopy and computed tomography (CT) scan of paranasal sinuses. Magnetic resonance imaging (MRI) was done in cases of suspected intracranial extension. After screening by routine mycological examination, the diagnosis was confirmed using complementary molecular methods. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the frozen-section biopsy were also compared with FFPE. Of the 200 suspect patients, 47 cases (23.5%) met the criteria for AIFRS. Species of the genus Aspergillus were the predominant 27 (57.4%) followed by Mucorales species 10 (21.3%), and Fusarium spp 3 (6.4%). Also, 3 cases (6.4%) of co-infection due to Aspergillus/Rhizopus were reported. The accuracy, sensitivity, specificity, PPV, and NPV of frozen section assessments were 99.5%, 97.9%, 100%, 100% and 99.3%, respectively. For GMS frozen-section alone, sensitivity, specificity, NPV, and PPV was 100%. Overall, the calculated accuracy of FFPE was 98.5%, sensitivity was 94%, specificity was 100%, PPV was 100%, and NPV was 98.1%. Examination of the frozen-section biopsy is a highly predictive tool for a rapid and effective diagnosis of patients with suspected AIFRS. We observed that GMS frozen-section is a fast and reliable exam to confirm the diagnosis of fungal invasion, with good accuracy, sensitivity, and specificity compared to the gold-standard FFPE biopsy.

Parasite-derived microRNAs as a diagnostic biomarker: potential roles, characteristics, and limitations.

MicroRNAs (miRNAs), a subclass of small regulatory RNAs that present from ancient unicellular protozoans to parasitic helminths and parasitic arthropods. MiRNAs' mode of action has attracted wide attention as a result of their unique functional importance. MiRNAs play a role in diverse physiological and pathological processes ranging from organ development, immune function to apoptosis and cancer at the post-transcription gene expression. Thus, miRNAs are known to be targets for clinical treatment and therapy. The discovery of the high stability of circulating miRNA in various types of host body fluids, such as whole blood, serum, plasma, saliva, and urine has increased great interest among researchers in the potential of circulating miRNA as a prognosis/diagnosis of infectious. Some circulating miRNAs biomarkers advanced to clinical applications related to human diseases. However, this idea starts to come only in the fields of infectious disease. The goal of this review is to enhance the current understanding of these molecules and their applicability in the field of medicine. A detailed review of the available literature consulting tools performed in online repositories such as NCBI, PubMed, Medline, ScienceDirect, and UpToDate. This review summarizes an overview of preclinical studies using circulating miRNAs biomarkers against infectious diseases affecting humans. The use of miRNA as a safe and potential tool is encouraging news, considering that until now, guidelines for the use of miRNA in clinical practice are still lacking.

Fungal contamination of indoor public swimming pools and their dominant physical and chemical properties.

Introduction: Considering the existence of both parasitic and fungal pathogens in the indoor public swimming pools and non-utilization of suitable filtration and disinfection systems in these places, this research aimed to determine the relationship between the indoor public swimming pools and possible pollution with parasitic and fungal agents, as well as physical and chemical characteristics of these pools and compare the results with national standards. Methods: In this study, 11 active indoor swimming pools of Zahedan city were sampled, using plastic pump techniques, from the middle of winter to the late summer season. A total of 88 water samples (eight water samples from each pool) were examined to determine the residual chlorine, contamination with parasitic and fungal agents, using culture media and slide culture techniques. Results were analyzed with SPSS software (V16) and, Microsoft Excel (V2010). Results: The findings revealed fungal contamination with Cladosporium, Penicillium, Aspergillus flavus and Aspergillus fumigatus, etc. and the physicochemical factors comply with the minimum standards, which indicates the need for continuous monitoring and control of water filtration and disinfection of water in the pools. Conclusion: The results show reasonable derangement of physicochemical and microbial factors of the evaluated pools. Efforts shall be made by the concerned authorities to provide health education to users, quality water at the pools and to maintain the safety and quality of the water through proper and adequate chlorination.

Distribution of invasive fungal infections: Molecular epidemiology, etiology, clinical conditions, diagnosis and risk factors: A 3-year experience with 490 patients under intensive care.

Recently, the prevalence of invasive fungal infections (IFIs) is rising. The global mortality rate of IFIs is 10-49%. This study aimed to determine the prevalence, the causative agents, and the risk factors associated with the invasive fungal infections in a tertiary health center to provide valid decision-grounds for healthcare professionals to effectively prevent, control, and treat fungal infections. The current study was conducted on 1477 patients suspected to have systemic fungal infections from different units of the hospital. After screening using routine mycological examination, the patients were confirmed with complementary mycological and molecular methods. Patients were included based on the confirmed diagnosis of IFI and excluded based on lack of a microbiologically and histologically proven diagnosis of IFI. Of the 1477 patients recruited in this study, confirmed cases of fungal infection were 490 (169 proven; 321 cases probable). Among the fungi recovered, Candida species had the highest frequency 337 (68.8%) followed by Aspergillus species 108 (22.1%), Zygomycetes species 21 (4.3%), non-Candida yeast 9 (1.8%). Others were black fungi 5 (1%), mycetoma agents 5 (1%), Fusarium 4 (0.8%), and Trichoderma (0.2%). Hematologic malignancies and diabetes mellitus were the most common underlying diseases among IFI-confirmed patients. This study observed an increased frequency of invasive candidiasis with non-albicans Candida and other invasive saprophytic fungal infections. The increased rate of invasive candidiasis with non-albicans agents highlights a new perspective in the epidemiology and treatment of invasive fungal infections.

Evaluation of phenotypes and genotypes of airborne Fungi during middle eastern dust storms.

Microbial species such as bacteria and fungi can be transported by dust storms over long distances, and may change the mycobiota in downwind. This study aimed to evaluate phenotypes and genotypes of airborne fungi during the Middle Eastern dust (MED) events and normal days in Khorramabad, Iran. The samples were collected regularly every six days at three locations during April 2018-March 2019, with additional samplings during MED days. For phenotypic analyses, the Petri dishes were incubated at 25 °C for 72-120 h. Molecular identification of fungi was carried out using polymerase chain reaction (PCR). The average (±SD) of total fungal concentration was 460.9 (±493.2) CFU/m3. The fungi with the highest average concentrations included Cladosporium cladosporioides, Penicillium brevicompactum, and Cladosporium iridis, respectively. The average concentration of fungi during dust days (967.65 CFU/m3) was 3.6 times higher than those in normal days (267.10 CFU/m3). During normal and dust days, 61 and 45 species were detected, respectively. Aspergillus and Cladosporium spp. were relatively more dominant during normal and dust days, respectively. Eight fungal species were only observed during MED days, including Talaromyces albobiverticillius that was detected for the first time in Iran. Despite air temperature, relative humidity and wind speed were associated to the fungal concentrations. Dust events lead to the changes in the air pollutants composition and mycobiota, identification of new fungi, and elevated fungal concentrations that may extremely affect the public health.

Increased Urine Interleukin-17 and Interleukin-22 Levels in Patients With Candidal Urinary Tract Infection.

INTRODUCTION: Candiduria is common in the hospitalized patients. This study aimed to quantify interleukin (IL)-17 and IL-22 levels in urine of candiduric patients. MATERIALS AND METHODS: A case-control study was conducted on inpatients at Hashemi Nejad Kidney Center. Thirty-four patients were identified with Candida species in their urine samples (> 103 colony-forming units per milliliter and presence of Candida species only). Urine samples with concomitant infections were excluded. Thirty-four patients with negative direct examination and culture were included as the control patients. Interleulin-17 and IL-22 levels were measured in the lyophilized and nonlyophilized urine. The relevant cytokine titers of the two groups were compared, and the association of cytokine elevation and candiduria was investigated. RESULTS: The majority of the candiduric patients were from the intensive care and urology units of women. Only 4 patients (11.7%) manifested fever and dysuria. Massive leukocyturia was observed in 4 patients. Candida glabrata was the most commonly isolated species (44%). Levels of the urine IL-17 and IL-22 were significantly elevated in the candiduric patients, when compared to the noncandiduric controls. While an increased IL-17 level was significantly associated with candiduria (odds ratio, 1.09; 95% confidence interval, 1.003 to 1.17; P = .04), an increased IL-22 level was not. The results showed that lyophilized urine samples maximized the detection power of urinary cytokines. CONCLUSIONS: Our results indicated that direct examination, fungal urine culture, and investigation of urine IL-17 and IL-22 levels are useful tools for diagnosis of Candida urinary tract infection.

The anti-giardial effectiveness of fungal and commercial chitosan against Giardia intestinalis cysts in vitro.

Chitosan with poly-N-acetylglucosamine sequences is a deacetylated derivative of chitin that can be found in the exoskeletons of crabs, shrimp and lobsters, the cuticles of insects and the cell walls of fungi. The aim of the present study was to compare the effects of fungal chitosan (FC) prepared from the cell walls of Penicillium viridicatum and Penicillium aurantiogriseum with commercially available chitosan (CC) against Giardia intestinalis cysts in vitro. The giardia cysts were isolated using a sucrose method. Four concentrations (50, 100, 200 and 400 µg/ml) of each type of prepared chitosan were applied for 10, 30, 60 and 180 min. The viability of the cysts was checked via 0.1 % eosin staining. Our results indicate that P. viridicatum (with a 47.5 % DD) and P. aurantiogriseum (with a 47.3 % DD) at different concentrations after 180 min precipitated, respectively, 56, 69, 81 and 100 %, and 63, 75, 86 and 100 % mortality rates. CC (with a 54 % DD) showed 79, 84, 93 and 100 % mortality rates. In conclusion, both FC and CC at 400 µg/ml concentrations after 180 min of exposure showed the most potent effect against G. intestinalis cysts. Accordingly, chitosan could be suggested as a new natural nanoform agent for future research in the safe and effective treatment of Giardia infections.

Chemical composition, protoscolicidal effects and acute toxicity of Pistacia atlantica Desf. fruit extract.

The present study was designed to evaluate the chemical composition and scolicidal effects of Pistacia atlantica Desf. extract against protoscoleces of hydatid cysts and its acute toxicity in mice model. Various concentrations of the methanolic extract (5-50 mg/mL) were used for 10-60 min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1%). Acute toxicity was also determined in mice model. The main components were ß-myrcene (41.4%), α-pinene (32.48%) and limonene (4.66%). Findings demonstrated that P. atlantica extract at the concentrations of 25 and 50 mg/mL after 20 and 10 min of exposure killed 100% protoscoleces. The LD50 of the intraperitoneal injection of the P. atlantica methanolic extract was 2.43 g/kg and the maximum non-fatal dose was 1.66 g/kg. Obtained results showed the potential of P. atlantica extract as a natural source with no significant toxicity for the production of new scolicidal agent to use in hydatid cyst surgery.