Direct Microscopic Examination Research Articles

Clinico-mycological analysis and antifungal resistance pattern in human immunodeficiency virus-associated candidiasis: An Indian perspective.

Objectives:Candidiasis is a common human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome-associated opportunistic mycoses. The present study ascertained the species spectrum of Candida strains recovered from different clinical samples from symptomatic HIV-positive individuals and determined the antifungal susceptibility profile of the isolates.Materials and Methods:A variety of specimens were collected from 234 symptomatic HIV seropositive individuals depending on their clinical manifestations and subjected to direct microscopic examination. Blood samples were inoculated in biphasic blood culture medium and all other specimens on Sabouraud dextrose agar with chloramphenicol and incubated at 35°C–37°C. Species identification of the recovered Candida isolates was attempted on the basis of germ tube production, micromorphology on corn meal agar, color and morphology on HiCrome Candida Differential agar, and carbohydrate fermentation and assimilation tests. Susceptibility testing of the isolates was performed employing the VITEK 2 system.Results:A total of 167 Candida isolates were obtained; Candida albicans (136), Candida tropicalis (13), Candida krusei (8), Candida parapsilosis (5), Candida glabrata (4), and Candida kefyr (1). Fluconazole resistance was more frequent among nonalbicans species, and significantly higher 5-fluorocytosine resistance compared to C. albicans was also observed. Eight Candida strains (six C. krusei, one C. kefyr, and one C. albicans) were multidrug resistant.Conclusion:Although C. albicans continues to be the leading etiological agent of candidiasis, the incidence of nonalbicans species among HIV-positive Indian individuals is rising. Antifungal resistance was higher among nonalbicans Candida species. Another issue of therapeutic concern is the possible emergence of multidrug-resistant Candida strains among these patients.

Schistosoma mansoni infection in human and nonhuman primates in selected areas of Oromia Regional State, Ethiopia.

The transmission of schistosomiasis, caused by trematodes of the genus Schistosoma, relies on freshwater snails that act as an intermediate host while human and other mammalian act as the definitive hosts. Many non-human primates (NHPs) such as Chlorocebus aethiops (vervet) and Papio anubis (baboon) are reported to be infected with Schistosoma mansoni in Ethiopia, but the role they play in parasite maintenance and transmission is still not clear. The objective of this study was, therefore, to determine the prevalence of S. mansoni infection in human and NHPs living in close proximities to villages in selected endemic areas of Ethiopia. In this cross-sectional study, stool specimens were collected from 911 humans, and fresh faecal droppings from 106 NHPs from Bochesa (Ziway), Bishan Gari (Kime) and Finchaa (Camp 7) endemic localities in Oromia Regional State, and examined for S. mansoni and other helminth infections using Kato-Katz method for human participants and direct microscopic examination for NHPs. The prevalence of helminthiasis among the human study population was 42.4% (386/911), and for soil-transmitted helminth infections (A. lumbricoides, hookworms, and T. trichiura) it was 13.4% (122/911). In humans S. mansoni was the predominant parasite, 23.1% (210/911) followed by A. lumbricoides, 8.7% (79/911); hookworms, 5.8% (53/911); T. trichiura, 4.8% (44/911); Taenia species, 2.2% (20/911); E. vermicularis, 2.1% (19/911); and H. nana, 3.2% (29/911). NHPs were found positive for Trichuris species and Strongyloides species besides S. mansoni. NHPs might play a significant role in local transmission and maintenance of S. mansoni infection even in the absence of human hosts. This calls for supplementation of chemotherapy for schistosomiasis along with measures such as snail control to interrupt transmission of the disease from humans to NHPs, and vice-versa.

A simple but yet novel stain in the diagnosis of onychomycosis

Background: Onychomycosis can resemble other nail dystrophies and hence the clinical diagnosis should be supplemented with a laboratory diagnosis. Rapid diagnosis of onychomycosis is the necessary to initiate early therapy. In this study, the efficacy of Chicago Sky Blue (CSB) stain in comparison with that of routine potassium hydroxide (KOH) preparation in the diagnosis of onychomycosis was evaluated. Methods: A prospective study of 42 nails with a clinical diagnosis of distal lateral subungual onychomycosis. KOH mount followed by direct microscopic examination of the subungual debris was performed. This was followed by microscopic examination after addition of 1% CSB. Results were interpreted by two experienced dermatologists. Results: 18 (42.9%) nails showed KOH positivity and 37 (88.1%) were positive for CSB stain. Conclusions: CSB staining was more sensitive and was more accurate in the rapid diagnosis of onychomycosis. Colour contrast provided by the CSB made it easier identification of fungal elements.

A prospective study of the epidemiological and clinical patterns of recurrent dermatophytosis at a tertiary care hospital in India.

Recurrent and clinically unresponsive dermatophytosis is being increasingly encountered in our country. It runs a protracted course with exacerbations and remissions. However, there is little information regarding the extent of the problem and the characteristics of recurrent dermatophytosis in published literature. We sought to determine the prevalence, risk factors and clinical patterns of recurrent dermatophytosis in our institution. We also investigated the causative dermatophyte species and antifungal susceptibility patterns in these species. One hundred and fifty patients with recurrent dermatophytosis attending the outpatient department of the Postgraduate Institute of Medical Education and Research, Chandigarh, India were enrolled in the study conducted from January 2015 to December 2015. A detailed history was obtained in all patients, who were then subjected to a clinical examination and investigations including a wet preparation for direct microscopic examination, fungal culture and antifungal susceptibility tests. Recurrent dermatophytosis was seen in 9.3% of all patients with dermatophytosis in our study. Trichophyton mentagrophytes was the most common species identified (36 patients, 40%) samples followed by T. rubrum (29 patients, 32.2%). In-vitro antifungal susceptibility testing showed that the range of minimum inhibitory concentrations (MIC) on was lowest for itraconazole (0.015-1), followed by terbinafine (0.015-16), fluconazole (0.03-32) and griseofulvin (0.5-128) in increasing order. A limitation of this study was the absence of a suitable control group (eg. patients with first episode of typical tinea). Recurrence of dermatophytosis was not explainable on the basis of a high (MIC) alone. Misuse of topical corticosteroids, a high number of familial contacts, poor compliance to treatment over periods of years, and various host factors, seem to have all contributed to this outbreak of dermatophytosis in India.

Laboratory Focus on Improving the Culture of Biosafety: Statewide Risk Assessment of Clinical Laboratories That Process Specimens for Microbiologic Analysis.

The Wisconsin State Laboratory of Hygiene challenged Wisconsin laboratories to examine their biosafety practices and improve their culture of biosafety. One hundred three clinical and public health laboratories completed a questionnaire-based, microbiology-focused biosafety risk assessment. Greater than 96% of the respondents performed activities related to specimen processing, direct microscopic examination, and rapid nonmolecular testing, while approximately 60% performed culture interpretation. Although they are important to the assessment of risk, data specific to patient occupation, symptoms, and travel history were often unavailable to the laboratory and, therefore, less contributory to a microbiology-focused biosafety risk assessment than information on the specimen source and test requisition. Over 88% of the respondents complied with more than three-quarters of the mitigation control measures listed in the survey. Facility assessment revealed that subsets of laboratories that claim biosafety level 1, 2, or 3 status did not possess all of the biosafety elements considered minimally standard for their respective classifications. Many laboratories reported being able to quickly correct the minor deficiencies identified. Task assessment identified deficiencies that trended higher within the general (not microbiology-specific) laboratory for core activities, such as packaging and shipping, direct microscopic examination, and culture modalities solely involving screens for organism growth. For traditional microbiology departments, opportunities for improvement in the cultivation and management of highly infectious agents, such as acid-fast bacilli and systemic fungi, were revealed. These results derived from a survey of a large cohort of small- and large-scale laboratories suggest the necessity for continued microbiology-based understanding of biosafety practices, vigilance toward biosafety, and enforcement of biosafety practices throughout the laboratory setting.

American tegumentary leishmaniasis: epidemiological and molecular characterization of prevalent Leishmania species in the State of Tocantins, Brazil, 2011-2015.

ABSTRACTDetermination of the epidemiological profile of the American tegumentary leishmaniasis (ATL) and identification of Leishmania species that are prevalent in the State of Tocantins were carried out through a retrospective and descriptive study based on data reported in SINAN, in the period from 2011 to 2015. Molecular techniques such as PCR-RFLP and PCR-G6PD to amplify Leishmania DNA were performed on stored on Giemsa-stained slides from lesion scarifications of ATL patients who were amastigote-positive by the direct microscopic examination. There were 1,434 ATL cases in Tocantins reported in this period. The highest incidence was reported in men aged over 60 years, rural residents, the most affected ethnic group was mixed ethnicity (mixed black and white) and the ones with lower education. The predominant clinical form was cutaneous, being diagnosed mainly by laboratory methods. Pentavalent antimonial was effective in resolving cases. The predominant species found in 271 analyzed samples from 32 municipalities located in 8 different health regions of Tocantins was Leishmania (Viannia) braziliensis. Identifying the epidemiological profile and characterizing the Leishmania spp species on regional level is essential to establish control and prevention behaviors, minimizing the number of cases and treatment resistance, recurrence and evolution to mucosal forms.

Association of Malassezia pachydermatis with the otitic and the normal ear canal of the dogs

The purpose of this study was to investigate the presence of M. pachydermatis in otitic and healthy ear canals of the dogs and to test the sensitivity of this microorganism to antifungal agents. A total of 180 swabs, 98 from otitic and 82 from clinically healthy dogs, were collected during the years 1998-2000 in Thessaloniki area (Greece). From all the swabs, smears for direct microscopic examination and inoculation on selective media for bacteria and fungi isolation were made. From the 90 M. pachydermatis isolates, 68 (69,38%) were made from infected and 32 (39,02%) from clinically healthy dogs. M. pachydermatiswas the sole isolate in 20 (20,39%) infected and in 12 (14,63%) non infected dogs. In 48 otitic and in 20 clinically healthy dogs, M. pachydermatis was associated with bacteria as Staphylococcus spp., Pseudomonas spp., Proteus spp. and Streptococcus spp. and fungi as Candida spp. ana Aspergillus spp.. S. intermediuswas isolated from 13 infected and 20 non infected animals. A mixed bacterial flora was grown from 6 infected and 22 clinically healthy animals, respectively, while no growth of microorganisms from 11 otitic and 8 healthy dogs was observed. All Malassezia tested strains (46) were found sensitive to ketoconazole, econazole, miconazole and clotrimazole. Nystatin was found effective to 38 isolates and noneffective to 8.

Prevalence and staining characteristics of Blastocystis isolates from food animals in Tamil Nadu.

The present study was undertaken to investigate the prevalence and staining characteristics of Blastocystis isolated from food animals. Smears of the duodenal and caecal mucosal scrapings, collected from food animals, were stained with Giemsa, Gram's, modified acid-fast and acridine orange. Blastocystis was identified in 295 samples, including faeces and intestinal contents of animals like small ruminants (95), poultry (170) and pigs (30). The prevalence in pigs was found to be high (94.4%) followed by poultry (29.4%) and small ruminants (14%). Various forms of Blastocystis such as vacuolar, granular and amoeboid forms were identified by using different stains. The parasites stained with Giemsa were identified by the presence of eosinophilic nucleus and basophilic cytoplasm. In organisms stained with Gram's stain, the cytoplasm of the vacuolar forms took up the counter stain safranine. Blastocystis appeared as a pink colored cyst against bluish green background with modified acid-fast staining. The study shows that there is a very high prevalence of Blastocystis among the food animals investigated. Simple parasitological procedures, including direct microscopical examination and staining with agents like Giemsa, Gram's and acridine orange can assist identification of the parasites from intestinal contents and faecal material.

Congenital Transmission of Trypanosoma cruzi in Argentina, Honduras, and Mexico: An Observational Prospective Study.

Compared with South America, there is a lack of epidemiologic studies about the risk of congenital transmission of Trypanosoma cruzi in Central America and Mexico. It has been suggested that T. cruzi genotypes might differ by region and that congenital transmission might vary according to the parasite's genotype. Our objective was to compare T. cruzi congenital transmission rates in three countries. We performed an observational prospective study in 2011-2014 enrolling women at delivery in one hospital in Argentina, two hospitals in Honduras, and two hospitals in Mexico. Congenital T. cruzi infection was defined as the presence of one or more of the following criteria: presence of parasites in cord blood (direct parasitological microscopic examination) with positive polymerase chain reaction (PCR) in cord blood, presence of parasites in infant's blood at 4-8 weeks (direct parasitological microscopic examination), and persistence of T. cruzi-specific antibodies at 10 months, as measured by at least two tests. Among 28,145 enrolled women, 347 had at least one antibody rapid test positive in cord blood and a positive enzyme-linked immunosorbent assay in maternal blood. PCR in maternal blood was positive in 73.2% of the cases, and genotyping identified a majority of non-TcI in the three countries. We found no (0.0%; 95% confidence interval [CI]: 0.0, 2.0) confirmed congenital case in Honduras. Congenital transmission was 6.6% (95% CI: 3.1, 12.2) in Argentina and 6.3% (95% CI: 0.8, 20.8) in Mexico. Trypanosoma cruzi non-TcI predominated and risks of congenital transmission were similar in Argentina and Mexico.

Sequence Base Identification of Respiratory Mucormycosis

Background: Mucormycosis is an uncommon fungal infection in immunocompromised patients during the past decades. Identification of causative agents could play an important role in the management of infected patients. Objectives: The aim of the present study was the identification of etiologic agents of respiratory tract mucormycosis, based on sequencing methods. Methods: Sinus tissue, bronchoalveolar lavage, and blood samples from the patients with suspected invasive fungal diseases were collected. Sinus tissue and bronchoalveolar lavage were examined by microscopic examination and cultured on Sabouraud dextrose agar. Blood samples were cultured on BACTEC medium. Semi-nested polymerase chain reaction (PCR) for diagnosis of mucormycosis was performed on samples, and products of positive PCR were sequenced and manually viewed with Chromas version 2.24 software. Pathology reports were collected from patients’ files. Results: Direct microscopic examinations, culture, and semi-nested PCR were positive in 11.7% (19/163), 6.7% (11/163), and 10.4% (17/163) of patients, respectively. None of the blood cultures were positive for Mucorales. The etiologic agents were Rhizopus oryzae (10 cases), R. microsporus (5 cases), and new species (2 cases). This new sequence (645 bp) was published in Gene bank and European Nucleotide archive of EMBL-EBI, and demonstrated 98% identity with Lichtheimia (Absidia) corymbifera genus. Conclusions: Management of mucormycosis has an important role in the treatment and outcome of such infections. Molecular assay and DNA sequencing could be used in parallel with conventional mycology techniques to identify Mucorales and for best management of respective infections.

Epidemiology, Causative Agents and Clinical Features of Onychomycosis in El-Gharbia Governorate

ONYCHOMYCOSIS is a fungal disease of the nails that infects the nail plate, nail bed and matrix, caused by various species of dermatophytes, yeasts and molds. The aim of this investigation is to determine the clinicomycological and epidemiological profile of the etiologic agents of onychomycosis. The study population included 100 suspected cases of onychomycosis referred to the outpatient clinic at the Department of Dermatology at Tanta University Hospital, El-Gharbia Governorate, Egypt during 2015-2016. Nail clippings were collected in sterile Petri dishes for direct microscopic examination and culture. Fungal isolates were identified. The results showed that candidal onychomychosis was the most common clinical type. Candida tropicalis was the most prevalent causative species. It was more common among females aged 41–50 years. The incidence was higher during summer and autumn seasons. Housewives were the commonest infected population. Fingernails were affected more frequently than toenails. The pattern of causative agents and clinical signs of onychomycosis is altering region to region, so repeated epidemiological surveys of onychomycosis seems to be fundamental. The present study provided novel and appropriate epidemiologic data of onychomycosis for the better prevention and treatment of this fungal infection.

Utility of a serum Aspergillusgalactomannan assay in diagnosis of invasive pulmonary Aspergillosis in HIV/AIDS patients: a prospective analysis

Background: Invasive pulmonary aspergillosis (IPA) is a life threatening infection among immunocompromised patients. A timely and precise diagnosis of this condition is often precluded by lack of sensitive and specific noninvasive diagnostic tools. Detection of galactomannan has emerged as a promising tool in this regard. While the analytical performance of serum galactomannan assay has been extensively evaluated in various high risk groups, its possible utility in HIV/AIDS patients is yet to be explored. The present study was undertaken to evaluate the yield of a serum galactomannan assay for diagnosis of IPA in a cohort of HIV/AIDS patients.Methods: 49 febrile HIV reactive patients with co-existent symptomatic AIDS were prospectively enrolled for the evaluation. Blood samples were drawn from each patient and serum galactomannan screening was performed employing the commercially available serum galactomannan assay, Platelia™ Aspergillus EIA. An optical density index >0.5 for at least two serum samples was considered the defining criteria for a positive result. Furthermore, sputum samples were collected and direct microscopic examination (10% potassium hydroxide wet mount) and fungal cultures were performed.Result: In accordance with the definitions laid down under EORTC/MSG (European Organization for Research and Treatment of Cancer / Mycoses Study Group) criteria, ten cases of probable and 23 cases of possible IPA were diagnosed. The sensitivity, specificity, and positive and negative predictive values of Platelia™ Aspergillus EIA for patients with probable IPA were 90%, 100%, 100% and 94.12 % respectively.Conclusion: Serum galactomannan assay has a high sensitivity and specificity and when complemented with relevant lower respiratory tract symptomatology and imaging findings, can serve as a useful adjunct to conventional modalities for the diagnosis of IPA in HIV/AIDS patients. DOI: 10.21276/APALM.1439

PCR-based detection of Aspergillus fumigatus and absence of azole resistance due to TR34 /L98H in a french multicenter cohort of 137 patients with fungal rhinosinusitis.

Fungal rhinosinusitis (FRS) has a worldwide distribution, comprises distinct clinical entities but is mostly due to Aspergillus among which Aspergillusfumigatus plays a major role in European countries. Although, there is accumulating evidence for the emergence of environmentally acquired-azole resistance in A.fumigatus (such as TR34 /L98H) in various clinical settings, there is few data for patients with FRS. In this study, we aimed to investigate the prevalence of A.fumigatus azole resistance due to TR34 /L98H in a multicentre cohort of patients with FRS. One hundred and thirty-seven patients with FRS admitted between 2002 and 2016 at four French medical centres were retrospectively enrolled. Clinical and mycological findings were collected. Aspergillus fumigatus and the TR34 /L98H alteration conferring azole resistance were investigated directly from clinical samples using the commercial CE-IVD marked MycoGENIE® A.fumigatus real-time PCR assay. Fungal ball was the more frequent clinical form (n=118). Despite the presence of fungal hyphae at direct microscopic examination, mycological cultures remained negative for 83 out of the 137 patients (60.6%). The PCR assay proved to be useful allowing the identification of A.fumigatus and etiological diagnosis in 106 patients (77.4%) compared with 44 patients (32.1%) when using culture as the reference method. Importantly, neither TR34 nor L98H alterations were evidenced.

Estudio epidemiológico de la tinea capitis en una población de Santiago de Chile. La aparición de Trichophyton tonsurans como agente etiológico

IntroductionTinea capitis (TC) is a dermatophytosis that affects the hair and scalp. The epidemiology of TC depends on the different geographical areas and is variable over time. Direct microscopic examination and fungal culture are essential to confirm the diagnostic suspicion and to identify the germ involved. ObjectivesTo determine the frequency of TC, to identify the etiological agents and to analyze the epidemiological aspects, in patients with suspected TC from Santiago Norte during the period 2009-2015. Materials and methodsA total of 505 patients with suspected TC were evaluated in the Dermatology Laboratory, Hospital Clinico Universidad de Chile. Direct microscopic examination was performed with KOH 30% and fungal culture of scalp lesions. Identification of the fungi was mainly morpho-physiological. ResultsThe diagnosis of TC was confirmed in 155 cases (30.7%). Male gender predominated (57.4%). The average age was 5years. 85.2% of the cases occurred in the preschool and school population. The most frequent isolated agent was Microsporum canis in 89.8%, followed by Trichophyton tonsurans in 7.1%. It was confirmed that 81.8% of the cases of Trichophyton tonsurans had foreign ancestry, mainly of Haiti and Peru. ConclusionsThis study evidenced the occurrence of Trichophyton tonsurans as a cause of TC, which could be explained by the increase in the immigrant population and the anthropophilic characteristics of the dermatophyte.

Profil mycologique des onychomycoses des mains chez les vendeurs de « Garba » à Abidjan (Côte d’Ivoire)

The aim of this study is to determine the prevalence and the fungi involved in onychomycosis located on fingers in Abidjan. This study includes the sellers of cooked cassava semolina served with fried tuna fish called "garba". Fines squames specimen was processed with 30% KOH for a direct microscope examination. Serous collection were done in physiological serum. Part of the original sample was cultured on Sabouraud medium with antibiotics (chloramphénicol, actidione). Species distribution and antifungal susceptibility by the technique on agar were noted. A total of 205 male sellers were involved. The Onychomycosis prevalence was 6.3%; (95% CI: 3.6-10.3). Five species of yeasts were identified. Candida spp were the most frequent (84.6%) and Candida albicans represented 30.8% of isolated yeasts. Trichosporon cutaneum were isolated in two case (15.4%). Susceptibility for amphotericin-B was 92.3% contrary to 5-fluorocytosine 30.8%. The subjects aged between 15 to 24 (69.2%), with a higher seniority (84.6%) and illiterate (69.2%) were the most affected. The main risk factors of fungal infections were the presence of previous fingernail onychomycosis (P<0.0001) and the frequent fingernail cutting (P=0.0009). Onychomycosis caused by yeast is infrequent among "garba" sellers in Abidjan. The main fungi involved in fingernail onychomycosis were Candida species. It is nevertheless necessary to implement sensitization for them on contributing factors.

Colonización por dermatofitos en conejos mantenidos en tiendas de mascotas de Santiago de Chile

Objetivo: Los dermatofitos son hongos queratinofílicos, de importancia en salud pública por su rol antropozoofílico. Dado el aumento en la adquisición de animales exóticos como mascotas y los escasos estudios sobre el estado de colonización por dermatofitos en estos animales, nos planteamos el objetivo de determinar la presencia de dermatofitos en conejos (Oryctolagus cuniculus) clínicamente sanos procedentes de tiendas de mascotas en Santiago de Chile. Materiales y Métodos: Se estudiaron 42 conejos clínicamente sanos fueron estudiados. Las muestras clínicas se obtuvieron fueron obtenidas desde el manto piloso y cultivadas en agar Sabouraud glucosado y dermatophyte test medium (DTM®). La identificación del agente aislado fue realizada por procedimientos micológicos clásicos que incluyeron examen microscópico directo y cultivo. Resultados: Del total de conejos estudiados, tres (7,1%) presentaron colonización por dermatofitos, siendo más frecuente en machos y en aquellos animales que convivían con cuyes. La especie identificada en los tres casos fue Trichophyton mentagrophytes. Conclusiones: Este estudio evidencia la colonización por dermatofitos en conejos domésticos, información importante para médicos veterinarios y propietarios de mascotas, al momento de estar en contacto con esta especie animal.

EFFICACY OF SIMPLE BLEACH DIGESTION OF SPUTUM IN THE DIAGNOSIS OF PULMONARY TUBERCULOSIS BY MICROSCOPIC EXAMINATION

Objective: Rapid diagnosis of pulmonary tuberculosis (PTB) depends on microscopic examination of sputum smear for acid-fast bacilli (AFB). The direct microscopy examination lacks sensitivity. It needs at least 10,000 AFB/ml of sputum to observe in the smear. The objective of this study was to determine the efficacy of three different short-term bleach digestions and concentration of sputum in the diagnosis of PTB.Methods: A total of 171 sputum samples collected from patients suspected of having pulmonary TB were used. Direct smear was prepared using each sample and then divided into three parts to which equal volume of 5% bleach was added. The first part was kept at room temperature for 45 minutes with intermittent shaking. The second part was allowed to stand at room temperature for 45 minutes for sedimentation, and the third part was kept at room temperature for 45 minutes and then centrifuged at 3000 g for 15 minutes for deposit formation. Smears were prepared and stained with Ziehl-Neelsen method and observed for AFB using Revised National TB Control Programme Guidelines.Results: Out of 171 sputum samples, 45 (26.32%), 48 (28.07%), 49 (28.65%), and 55 (32.16%) of specimens were positive by direct, bleach digested mixed method, bleach digested sedimented method, and centrifugation method, respectively. The background of the bleach digested sputum smears was clear.Conclusion: Short-term bleach digestion of sputum improves the sensitivity of microscopic examination and the number of AFB observed in the smear. Bleach being a disinfectant kills AFB and makes handling of sputum safer.

Direct identification of molds by sequence analysis in fungal chronic rhinosinusitis

Fungi are widely implicated in chronic rhinosinusitis. Direct microscopic examination (DME) is used to confirm the biological diagnosis of fungal rhinosinusitis (FRS). Diagnostic sensitivity of DME is better than culture, however DME does not allow fungal species identification. In this study, we included 54 sinus samples demonstrating hyphae on DME. Direct sequencing was compared to culture for the identification of the fungal species. Sequence analysis identified fungi in 81.5% of cases while culture was positive in only 31.5%. The most common genus was Aspergillus and the identified species belonged to section Fumigati or to section Flavi. Among other fungi identified by sequence analysis, Schizophyllum commune was present in three samples attesting to the importance of this Basidiomycetes in FRS. Our results clearly demonstrate the superiority of sequencing compared to culture when performed on specimens with hyphal elements at DME, and contributes to the epidemiological knowledge of fungi involved in FRS.

Diagnosis of coccidioidomycosis in a non-endemic area: Inference of the probable geographic area of an infection

BackgroundCoccidioidomycosis is one of the most important endemic mycoses in Northern Mexico. However, diagnosing this disease can be challenging, particularly in patients who do not reside in endemic areas. Case reportThe case of a Mexican HIV+ patient who developed fever, general malaise, a severe cough, and dyspnea during a stay in Acapulco, Guerrero, Mexico, is presented. Since various diseases are endemic to the state of Guerrero, the doctors originally suspected that the patient had contracted influenza A (H1N1), Q fever, or tuberculosis. All the diagnostic tests for those diseases were negative. The patient had received numerous mosquito bites while staying in Acapulco, and a nodule had appeared on his right cheek. Therefore, malaria, cryptococcosis, and histoplasmosis were also suspected, but those infections were also ruled out through diagnostic tests. A direct microscopic examination was performed using KOH on a sample taken from the cheek nodule. The observation of spherules suggested the presence of a species of Coccidioides. The fungus was isolated, and its identity was confirmed by phenotypic and molecular methods. The geographic area in which the infection was likely acquired was identified by random amplified polymorphic DNA (RAPD) analysis. The results suggested a probable endogenous reactivation. ConclusionsThis clinical case illustrates the difficulties associated with diagnosing coccidioidomycosis in non-endemic areas.

Cutaneous phaeohyphomycosis of foot web by Curvularia lunata

A case of cutaneous phaeohyphomycosis of the foot affecting interdigital spaces between toes in a 31 year old immunocompetent male with no history of diabetes is illustrated. Fugal elements were found in direct microscopic examination of skin scrapping.