Mycological Culture Research Articles

Micobacteriose cutânea atípica felina autóctone no município do Rio de Janeiro-Brasil

Background : : : Cutaneous mycobacterioses show a worldwide distribution and manifest as three forms: atypical mycobacteriosis, feline leprosy and cutaneous tuberculosis. Atypical cutaneous mycobacteriosis in cats is a rare disease commonly caused by fast-growing mycobacteria belonging group IV of the Runyon classification. This report describes for the first time an autochthonous case of atypical cutaneous mycobacteriosis in a cat from the municipality of Rio de Janeiro. Case : A castrated female mongrel cat presented multiple ulcerated, exudative and alopecic lesions in different regions of the body. After sedation of the animal with 1% acepromazine hydrochloride (0.1 mg/kg) and 10% ketamine hydrochloride (10 mg/kg), clinical examination was performed and an exudate was collected from the ulcerated skin lesion for cytopathological analysis and mycological culture. The slide containing the lesion impression was stained by a rapid panoptic method. For fungal culture, material was collected with a sterile swab, seeded onto Sabouraud dextrose agar containing chloramphenicol and mycobiotic agar, and incubated at 25oC. After antisepsis and asepsis, local anesthesia was performed with 2% lidocaine hydrochloride without vasoconstrictor and a 4 mm punch biopsy was collected from the lesion, fixed in 10% buffered formalin and sent for histopathological analysis. No fungal structures were detected by cytopathological or mycological analysis. Histopathological examination revealed ulcerated skin and the presence of a marked and diffuse pyogranulomatous infiltrate in the superficial and deep dermis, which also involving the subcutaneous tissue and underlying muscle tissue. Grocott’s or PAS staining did not identify fungal structures. The Wade method was positive for acid-fast bacteria suggestive of ycobacterium . Analysis of the hematological and biochemical profiles revealed no noteworthy alterations. After the result of the histopathological exam, exudate material and lesion fragments were collected for bacteriological examination. The clinical specimens were processed and the presence of acid-fast bacteria was confirmed by Ziehl-Neelsen staining. Mycobacterium fortuitum fortuitum was identified based on the Runyon classification and biochemical tests, and was confirmed by molecular tests. Oral administration of enrofloxacin (5 mg/kg) at 24 h intervals was prescribed for 30 days. On the control visit, involution of the skin lesions was observed by physical examination and was also reported by the owner. After 4 months of irregular treatment due to the difficulty in administering the medication because of the cat’s aggressiveness, the animal started to present respiratory difficulties, rapid weight loss, apathy and died. Discussion : The occurrence of atypical mycobacteriosis is rare in Brazil, with isolated reports of mycobacteriosis such as canine leproid granuloma syndrome in Sao Paulo, canine cutaneous mycobacteriosis in Rio Grande do Sul, one case of feline leprosy in Sao Paulo, and one case of feline atypical cutaneous mycobacteriosis in Sao Paulo. The diagnosis and treatment of mycobacterial infections are frequently neglected because they are not included in the differential diagnosis of diverse cutaneous infections. In view of the scarcity of reports and to alert veterinarians that the disease should be included in the differential diagnosis with other feline dermatopathies, this report described a case of atypical feline cutaneous mycobacteriosis that occurred in the state of Rio de Janeiro. In contrast to the previous case report in Sao Paulo, in which was diagnosed the species Mycobacterium fortuitum biovar peregrinum , the species identified here was Mycobacterium fortuitum biovar fortuitum .

Systemic Cryptococcosis in a Miniature Schnauzer Dog

Background: Cryptococcosis is an important zoonotic disease that occurs worldwide. The disease is caused by a soilborne opportunistic fungus of the genus Cryptococcus, which can also be found in the feces of birds, especially pigeons. In Brazil, the geographical distribution of the agent is fairly defined, with the species C. gattii predominantly found in thenorth-eastern states. Diagnosis is based on the clinical history, physical examination findings, and results of complementary testing, such as cytopathological and histopathological examinations. This report aims to describe the clinical and anatomopathological findings in a case of systemic cryptococcosis in a dog.Case: A 4-year-old female Miniature Schnauzer was referred for necropsy. For 3 months prior to death, the dog had displayed generalized alopecia, pruritus, and severe weight loss, Initially, a private veterinarian had suspected leishmaniasis. Gross examination was performed and samples for cytopathology, histopathology, and fungal culture were obtained. Themacroscopic examination revealed generalized alopecia, congested mucosa, cachexia, hyperemia of the brain, and multiple white nodules measuring 0.5 cm to 4 cm in the lungs and the kidneys. None of the other organs showed significant lesions. Cytopathological examination of the nodules revealed a highly cellular sample, with a severe chronic inflammatory response, characterized by multinucleated giant cells and round-to-elliptical, yeast-shaped structures (5-10 μm), suggestive ofCryptococcus organisms. Histopathological examination of the lungs, kidneys, and brain revealed a severe diffuse chronic inflammatory process, with lymphocytic infiltration and multinucleated giant cells; countless yeast-shaped, round-to-ovoid structures (similar in appearance to “soap bubbles”) that were negative in hematoxylin-eosin stain were also present.Grocott’s methenamine silver stain was then applied, which positively stained the organism capsules black, confirming Cryptococcus. For mycological diagnosis, samples from the lungs nodules were cultured on Sabouraud’s dextrose agar supplemented with chloramphenicol 0.4% and incubated for 7 days at 25-30ºC. Abundant small, smooth, irregular-sized,cream-colored colonies were obtained, and a mycological smear, stained by India ink, was performed.Discussion: Considered an uncommon disease, canine cryptococcosis can be misdiagnosed. Clinical findings, such as alopecia, lymphadenopathy, and cutaneous nodules that may be ulcerated, are commonly seen in these patients due to the hematogenous spread of the organism. In the case described, the dog’s owner was contacted and asked to provide epidemiological information. The owner reported that the dog had lived in an apartment, with little access to the street; however, pigeons were frequently observed on the balcony, along with pigeon feces in the dog feeder. The presence of pigeon feces is a risk factor for the development and spread of the fungus, which may be problematic to public health. Cytopathological examination is a simple, quick diagnostic tool with good sensitivity for some infectious agents. Mycological culture can provide results due to the characteristic of the Cryptococcus sp., since the encapsulated yeast produces a mucous-like colony. Cryptococcosis has a relevant role in public health, so a precise diagnosis is imperative. Its diagnosis is based upon culture, cytology and serological tools. When the animal does not survive to the ilness, the association between necropsyfindings and histopathological examinations is essencial.Keywords: canine, diagnostic, yeast, anatomopathology.

Dermatophytosis Caused by Microsporum canis in a Free-Living Maned Wolf (Chrysocyon brachyurus).

Background: Chrysocyon brachyurus is a South American wild canid considered a species near threatened by the International Union for Conservation of Nature and is classified as vulnerable in the National List of Endangered Species. With the increase of the contact between domestic animals, human population and wild animals, there was a greater exposure of the maned wolf to pathogens. Due to the importance of its conservation, the knowledge of emerging infectious diseases that affect this species becomes essential. This report aims to describe the first diagnosed case of dermatophytosis caused by Microsporum canis in a maned wolf.Case: A free-living female maned wolf (Chrysocyon brachyurus), approximately 60 days old, was rescued with presence of alopecia, non-pruritic lesions, one of circular shape located in the nasal plane and the other with signs of scaling and crusts in the region of the left pina. The animal was active, in good general condition and without other significantchanges to clinical examination. Skin scraping was performed for mycological culture of both lesions. Fungal growth on Sabouraud’s agar identified Microsporum canis. Topical therapy with ketoconazole ointment and cleaning of lesions with 0.2% chlorhexidine was instituted. After 20 days of treatment, remission of clinical signs and repilation of affected areas were observed. New mycological cultures of both areas were carried out, which were negative for Microsporum canis.Discussion: Although Microsporum canis is described as causing dermatophytosis in several animal species, it has apparently not yet been reported in maned wolves. Microsporum canis is one of the most isolated zoophilic dermatophytes in domestic cats and is also cited in reports of symptomatic wild felids, such as tigers, in which it has been identified as either the only agent of infection or in association with Trichophyton mentagrophytes. The occurrence is also high in domestic dogs. In the wild canids, Microsporum gypseum has been described in the gray wolf, Trichophyton mentagrophytes in wild foxes, and Microsporum canis in red fox, among others. The finding of zoophilic and geophilic dermatophyte fungi in healthy, free-living and captive wild animals confirms their role as carriers, probable reservoirs and disseminators of these organisms in the environment, and the potential risk they represent as causes of zoonoses. Zoophilic dermatophytes are usually transmitted by contact between individuals and contaminated fomites. In the case of M. canis in the red fox the contact was an asymptomatic domestic cat. In the present case the transmission was not determined, since the animal was rescued from the wild. Regarding the age group, young animals are more susceptible to dermatophytes than adults, probably related to the greater immunity of adults due to previous contacts or even the immaturity of the immune system of the young. The dermatophytosis diagnosis is generally based on anamnesis, physical examination of the lesion, Wood’s lamp examination, microscopic skin scraping, fungal culture, or even histology and PCR tests. In the present case, the final diagnosis was based upon by fungal culture in Sabouraud agar, which allowed to identify the dermatophyte species and,thus, the possible source of infection. Dermatophyte infections can be treated with systemic or topical antifungal medications. Because it was a young animal, it was chosen in the present case only for topical use, which proved to be adequate, probably due to the action of the drug in the areas of alopecia skin and the improvement of the systemic condition of theanimal. Knowing that the maned wolf is considered as a vulnerable species, it is important to know the diseases that affect this species, in order to carry out, when necessary, disease monitoring programs, preventive and therapies, which is essential for its preservation.Keywords: wild animal, disease, skin, dermatophyte.

Feather Loss in a Budgie

A 3 year-old budgie with the complaint of feather loss and disturbed body growth, was referred to the clinic. In history, it was understood that two offspring showed healthy growth within the first year of their life, then scaling on the skin was observed in one of the budgie’s feet. Over the next 3 months the body feathers began to fall. In the hairless regions, new feathers appeared in a cottony look and weak structure. But these new feathers shed before they could finish growing. The food and water appetite had not deteriorated during this time. In physical examination, there were a few feather in cotton appearance on the body of the budgie. Bird had difficulty in balancing his body while standing and walking. Nowhere in the bird's body has skin lesions. In mycological culture, Aspergillus spp was isolated. Bird died a week after being brought to the clinic. When the etiology of the disease is assessed in the budgies >3 years old, all feathers have fallen and body development has deteriorated, it has been concluded that it is appropriate to consider chronic diseases which cause weakening of the immune system resulting in diseases such as aspergillosis.

Mucormycosis: Battle with the Deadly Enemy over a Five-Year Period in India.

Mucormycosis is an emerging opportunistic fungal infection. Increasing immunocompromization, widespread use of antibacterial and antifungal agents (such as voriconazole prophylaxis), carcinomas, transplantation and lifestyle diseases such as diabetes are the main contributors to this situation. The predominant clinical manifestations of mucormycosis vary from host to host, with rhino-orbital-cerebral, pulmonary, cutaneous, and gastrointestinal infections being the most common. In India, the prevalence of mucormycosis is approximately 0.14 cases/1000 population, which is about 70 times the worldwide-estimated rate for mucormycosis. The present study was undertaken over a period of five years (January 2009–December 2014) to determine the prevalence of mucormycosis. The samples suspected of mucormycosis were examined by direct KOH wet mount and cultured on Sabouraud’s dextrose agar without actidione and on blood agar as per standard mycological techniques. Histopathological correlation was done for most of the cases. Antifungal susceptibility testing was performed by the EUCAST reference method. We identified a total of 82 cases of mucormycosis out of a total of 6365 samples received for mycological culture and examination during the said time period. Out of these, 56 were male patients and 27 were females. Most common presentation was rhino-orbito-cerebral (37), followed by cutaneous (25), pulmonary (14), oral cavity involvement (4) and gastrointestinal (2). The most common risk factors were diabetes and intramuscular injections. The fungi isolated were Rhizopus arrhizus (17), Apophysomyces variabilis (12), R. microsporus (9), Lichtheimia ramosa (8), Saksenaea erythrospora (5), Syncephalastrum racemosus (4), R. homothallicus (2), Rhizomucor pusillus (1), Mucor irregularis (1) and A. elegans (1). The mainstay of the treatment was amphotericin B, along with extensive surgical debridement whenever feasible. Most of the patients (50) recovered, but 25 died. The rest of the patients left against medical advice. “Nip in the Bud” should be the mantra for clinicians/surgeons for a favorable prognosis. Early diagnosis, prompt institution of appropriate antifungal therapy, surgical debridement whenever necessary, knowledge of risk factors and their timely reversal is the key for management.

Esporotricose conjuntival felina

Sporotrichosis is a zoonosis caused by the dimorphic fungus Sporothrix schenckii, which domestic cat is described as an important font of infection. Due to the lack of studies over ocular alterations that occur in feline sporotrichosis, this paper describes a case of ophthalmic alteration in a cat from the metropolitan area of Rio de Janeiro city without antecedent ocular injury history. Complete clinical examination was performed, verifying ulcerated cutaneous lesions and conjunctivitis. Material from a cutaneous lesion and from the left eyeball conjunctival sac was collected for citopathologic exam and mycologic culture, the diagnostic was confirmed through culture isolation of Sporothrix schenckii, from both the cutaneous and ocular lesions. Sporotrichosis treatment was started and 75 days after, the cutaneous lesions were healed and there was no sign of conjunctival alteration.

Performance of Molecular Approaches for Aspergillus Detection and Azole Resistance Surveillance in Cystic Fibrosis.

Aspergillus fumigatus triazole resistance is an emerging concern for treating chronically infected/colonized patients. This study sought to evaluate the performance of PCR assays to detect Aspergillus fungi together with azole resistance in sputum samples from cystic fibrosis (CF) patients. In total, 119 sputum samples from 87 CF patients were prospectively processed for Aspergillus detection by means of mycological culture and four qPCR assays, 2 in-house methods and two commercial multiplex real-time PCR assays simultaneously detecting Aspergillus and the most relevant cyp51A gene mutations (MycoGENIE® and AsperGenius®). Azole susceptibility of A. fumigatus isolates was assessed using Etest® method and cyp51A gene mutation were characterized by sequencing. The overall rate of Aspergillus detection with the four qPCR assays ranged from 47.9 to 57.1%, contrasting with 42/119 (35.3%) positive cultures with A. fumigatus. The high sensitivity of PCR on sputum could then contribute to more effective grading of Aspergillus disease in CF patients. Five out of 41 isolated strains (12.2%) exhibited azole-resistant MIC patterns, three of which harbored cyp51A mutations and only 1/3 with the sequence TR34/L98H. Combined with culture, PCR assay achieved high sensitivity Aspergillus screening in CF samples. However, cyp51A targeting was only moderately effective for azole resistance monitoring, while Aspergillus resistance remains of great concern.

Rhino-orbito-cerebral mucormycosis from dental origin: Case report

Rhino-orbito-cerebral mucormycosis from dental origin is an acute infection caused by opportunistic fungi belonging to the order of Mucorales, which affects mainly diabetic and immunocompromised patients.We report the case of a 63-year old diabetic man who performed a dental extraction on himself by his own means and subsequently developed a rhino-orbito-cerebral mucormycosis with cutaneous and palatal affection. The species isolated in the mycological culture was Rhizopus sp.

Tinea capitis: correlation of clinical presentations to agents identified in mycological culture

Tinea capitis: correlation of clinical presentations to agents identified in mycological culture

New evidence on oral L. plantarum P17630 product in women with history of recurrent vulvovaginal candidiasis (RVVC): a randomized double-blind placebo-controlled study.

To evaluate activity and tolerability of an oral product containing the specific probiotic strain Lactobacillus plantarum P17630 on vaginal colonization of lactic acid bacteria. L. plantarum P17630 (5x109 CFU/capsule) or placebo were formulated in capsules and administrated orally to a total of 93 women with a history of recurrent vulvovaginal candidiasis over a period of 3 treatment cycles (15 days/cycle) separated by 15-day wash-out intervals. Vulvovaginal examinations and vaginal mycological culture, including Nugent score and Lactobacillary grade (LBG), were assessed at baseline, at day 45 and at day 90 (end of the study). Probiotic intake significantly improved lactobacilli colonization on vaginal epithelial cells. The results demonstrated a significant difference in LBG score when comparing day 0 to day 45 (p=0.000016) and to day 90 (p=0.001415) in women treated with L. plantarum P17630. No evidence of improvement in LBG score was recorded in women who received placebo (p<0.005). The increased lactobacilli adhesion was associated with the improvement of clinical signs such as redness, swelling, and discharge. This study demonstrates that the administration of oral probiotic Lactobacillus plantarum P17630 improves vagina colonization of acid lactic bacteria (an increase of vaginal LBG score) and suggests the use of this oral product to successfully prevent episodes of vulvovaginal candidiasis. The clinical study described in this article is registered in the ISRCTN registry with trial registration ID ISRCTN12669692.

Suspected toxicity by biological waste and air sac nematode infestation in a free-living peregrine falcon (Falco peregrinus)

This case report describes an uncommon complex of mortality in a free living peregrine falcon (Falco peregrinus). The bird was found rinsed with biologic waste and the integument was mildly inflamed. No respiratory signs were detected at first. In addition, no alterations were detected during. During orthopaedic, neurologic, ophthalmologic and radiographic examinations. Parasitological pharyngeal and faecal examination revealed Capillaria spp. and Serratospiculum spp. eggs. The therapeutic scheme included stress reduction, supportive therapy with fluids, antifungals, antibiotics, anti-inflammatory, antiparasitic agents and nutritional management. Cleaning with dishwashing gel to remove the biological waste took place, but its removal was partial. Despite the intensive care, the patient developed respiratory distress and succumbed on the fifth day of hospitalization. Pathological examination revealed adult Serratospiculum tendo parasites in the air sacs and Capillaria spp. in the large intestine. Bacteriological and mycological cultures revealed significant growth of Klebsiella pneumoniae isolated from the skin, heart, lung, liver, kidney and intestine and a mild presence of Candida albicans in the large intestine. Their probable role was judged as secondary. Mild multifocal haemorrhages with heterophilic and lymphohistiocytic infiltrates, fibrin and rod‑like bacteria were detected in the skin histopathologically, confirming the clinical integument inflammation. Cause of death was asphyxia caused by oedema, haemorrhages and blood congestion of the lung. The contribution of S. tendo in the initial spill event and case evolution is discussed. This unusual case highlights the complexity of the wildlife cases, with which avian or small animal clinicians (consulting rehabilitation centres or falconers) might be confronted. Although the application of ancillary and advanced diagnostic tools, the in-depth knowledge of the species and the intensive monitoring are important, this might lead to an unrewarding outcome. Financial constraints in wildlife casualty cases urge the critical choice of diagnostic, therapeutic and post-mortem laboratory testing. Serratospiculum tendo should be considered in the differential diagnosis of acute respiratory distress in free-living peregrine falcons, presented in rehabilitation centres or in private practise by falconers in Greece and Europe.

Dermatophytes in Cats without Dermatopathies in the Metropolitan Area of Florianópolis, Brazil

Background: Dermatophytes are infectious agents responsible for dermatophytosis, an important worldwide zoonosis. Cats are considered potential hosts and reservoir of these fungi, especially Microsporum canis. The prevalence in cats without dermatopathies varies according to the region, climate and animal husbandry. The aim of this study was to estimate the frequency of dermatophytes in cats without clinical signs of dermatopathy in the Metropolitan Area of Florianópolis, situated in the coast of Southern Brazil.Materials, Methods &amp; Results: A total of 198 samples were obtained from cats without dermatopathies domiciled in the metropolitan area of Florianópolis. The collections were made through vigorous hair brushing throughout the body of the animal, using a sterile toothbrush. Mycological culture was performed onto Sabouraud Agar Chloramphenicol-Cyclohexamide (SCC), and incubated at 25-27°C for 21-28 days. The diagnosis was based on the macro and micromorphological characteristics of the isolated dermatophyte. One hundred and ten samples (55.6%) were collected in veterinary clinics and 88 (44.4%) in multiple household cats (average 11). The frequency of dermatophytes corresponded to 3.0% (6/198). Only the genus Microsporum was observed with predominance of M. canis (66.7%), followed by M. gypseum (33.3%). Saprotrophic fungi were observed in 94.4% of the cultures and 5.6% of the samples did not occurred fungal growth. Most of the isolates were obtained from adult cats (66.7%), females (83.3%) and with long hair (5.4%) in comparison to short hair samples (2.1%). Thirty percent of the cats (59/198) had been tested for retroviruses, and, among them, 27.0% were positive (22% FeLV and 5% FIV). M. gypseum was isolated from one feline FeLV positive. Various saprotrophic species were isolated from multiple household cats.Discussion: Dermatophytosis is considered as the most common occupational zoonosis among veterinarians, and it is recognized as a biological risk associated to the direct contact with cats. There are reports of regional outbreaks in Brazil where the pets were indicated as sources of dermatophytes to humans in the domestic environment. The frequency of dermatophytes observed in this study (3.0%) was lower than shown in others papers. Microsporum canis is commonly isolated from cats in hot weather regions and crowded living facilities; once introduced in to the creation on a cattery it is difficult to be eliminated. There is evidence of a higher occurrence of M. canis in cats FIV positive than seronegative ones. Dermatophytes were isolated from cats in contact with other cats (5/165). However, multiple household cats had no positive cultures and this outcome can be associated to high contamination by saprotrophic fungi. For diagnosis of dermatophytosis, mycological culture is indicated as the most efficient and easy to perform method, but it is necessary to be aware of false negative results, since it can occur in situations of intense contamination. Thus, it is advisable to prophylactically perform a clinical examination as a routine and also laboratory analyses in each cat before introducing it to a new environment.

Improving the diagnosis of invasive aspergillosis by the detection of Aspergillus in broncho-alveolar lavage fluid: Comparison of non-culture-based assays

ObjectivesThis study aimed to evaluate new tools to diagnose invasive aspergillosis (IA) directly from broncho-alveolar lavage (BAL) samples. MethodsAll consecutive patients with suspected IA who underwent bronchoscopy with BAL were prospectively included. Mycological culture and ELISA detection of galactomannan (GM) were performed on BAL. Two in-house and two marketed PCR assays were used on BAL DNA extracts to detect Aspergillus species. Susceptibility testing was performed after culture; marketed PCR assays detected mutations in the CYP51A gene associated to resistance. ResultsWithin 3 years, 1555 BAL samples were processed, including 413 samples from 387 immunosuppressed patients. IA diagnosis was no-IA, possible, probable or proven IA in 326, 23, 37 and 1 patients, respectively. PCR assays sensitivity for Aspergillus detection ranged from 61% to 74%, below GM (87%), but contrasting with 47% for cultures. Combining PCR to EORTC/MSG criteria increased the sensitivity to 100%. Interestingly, tests performance in non-hematological patients ranged from 60% to 75%, and were higher than in hematological patients, and those with prior exposure to antifungals. All 16 isolates of Aspergillus fumigatus were susceptible; PCR did not detect any resistance marker in the 37 A. fumigatus PCR-positive samples. ConclusionThe molecular detection of Aspergillus directly in BAL samples greatly improved the diagnosis of IA, particularly in non-hematological patients.

Mixed Infection of Respiratory Tract in a Dog Caused by Rhodotorula mucilaginosa and Trichosporon jirovecii: A Case Report

This report describes the isolation of two environmental fungi: Rhodotorula mucilaginosa and Trichosporon jirovecii accompanied by Pseudomonas aeruginosa and Escherichia coli from a dog with bronchotracheitis. All microorganisms were isolated routinely from a mucopurulent discharge, obtained during bronchoscopy from laryngotracheal area. The initial identification of yeasts was confirmed by API Candida and by molecular analysis of internal transcribed spacer region. Administered antimicrobial treatment with Marbofloxacin and Canizol has brought the improvement in the dogs’ health status. The final results of control mycological culture were negative. Most probably underlying hypothyroidism and the use of steroids were the factors predisposing this patient to opportunistic infection of mixed aetiology. As far as we are concerned, this is the first case of dogs’ respiratory tract infection caused by R. mucilaginosa and T. jirovecii.

Sarcoid-like paracoccidioidomycosis: case report

Paracoccidioidomycosis (PCM), which is caused by the dimorphic fungus Paracoccidioides spp, is the most prevalent systemic mycosis in immunocompetent individuals in Brazil. The fungus lives in the soil and usually infects the patient through inhalation of airborne propagules, the lungs being the portal of entry. In areas of larger endemicity, such as Brazil, the annual incidence reaches 3 cases per 100,000 inhabitants. Cutaneous lesions in PCM are frequent and ulcer or ulcerous-vegetative lesions are the commonest morphological type, followed by the infiltrative pattern. The histology shows a dermal granulomatous inflammation. Infiltrative PCM can be easily misdiagnosed, once clinical and histological similarities may occur specially with cutaneous sarcoidosis. We describe a case of cutaneous paracoccidioidomycosis misdiagnosed as sarcoidosis both in clinical examination and in the histopathological analysis. Further exams confirmed Paracoccidioides sp. as the causative agent through mycological cultures and Grocott’s stain. We believe this case may help physicians around the world to suspect and consider PCM as a differential diagnosis, especially if the patient has traveled to endemic area or the current therapy for sarcoidosis lacks clinical improvement.

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.

Experience with Saccharomyces boulardii Probiotic in Oncohaematological Patients

Very few reports have been published to date on the bloodstream infections caused by Saccharomyces spp. in oncohaematological patients, and there are no guidelines on the use of this probiotic microorganism in this population. We describe the use of probiotic preparation containing Saccharomyces boulardii in a large group of oncohaematological patients. We retrospectively analysed the data from 32,000 patient hospitalisations at the haematological centre during 2011–2013 (including 196 haematopoietic stem cell transplant recipients) in a tertiary care university-affiliated hospital. During the study period, 2270 doses of Saccharomyces boulardii probiotic were administered to the oncohaematological patients. In total, 2816 mycological cultures were performed, out of which 772 (27.4%) were positive, with 52 indicating digestive tract colonisation by Saccharomyces spp., mainly in patients with acute myeloid leukaemia (AML), myelodysplastic syndrome (MDS) or multiple myeloma (MM). While colonised, they were hospitalised for 1683 days and 416 microbiological cultures of their clinical samples were performed. In the studied group of patients, there were six blood cultures positive for fungi; however, they comprised Candida species: two C. glabrata, one C. albicans, one C. krusei, one C. tropicalis and one C. parapsilosis. There was no blood culture positive for Saccharomyces spp. Our study indicates that despite colonisation of many oncohaematological patients with Saccharomyces spp., there were no cases of fungal sepsis caused by this species.

Case Report: Histoplasmosis in Himachal Pradesh (India): An Emerging Endemic Focus

We describe four cases of histoplasmosis indigenous to Himachal Pradesh (India) that will be of considerable public health interest. A 48-year-old human immunodeficiency virus (HIV)-negative man with cervical and mediastinal lymphadenopathy, hepatosplenomegaly, adrenal mass, and bone marrow involvement was treated as disseminated tuberculosis without benefit. Progressive disseminated histoplasmosis was diagnosed from the fungus in smears from adrenal mass. Another 37-year-old HIV-positive man was on treatment of suspected pulmonary tuberculosis. He developed numerous erythema nodosum leprosum-like mucocutanous lesions accompanied by fever, generalized lymphadenopathy, and weight loss. Pulmonary histoplasmosis with cutaneous dissemination was diagnosed when skin lesions showed the fungus in smears, histopathology, and mycologic culture. Both were successfully treated with amphotericin B/itraconazole. Third patient, a 46-year-old HIV-negative man, had oropharyngeal lesions, cervical lymphadenopathy, intermittent fever, hepatosplenomegaly, and deteriorating general health. Progressive disseminated oropharyngeal histoplasmosis was diagnosed from the fungus in smears and mycologic cultures from oropharyngeal lesions and cervical lymph node aspirates. He died despite initiating treatment with oral itraconazole. Another 32-year-old man 3 months after roadside trauma developed a large ulcer with exuberant granulation tissue over left thigh without evidence of immunosuppression/systemic involvement. He was treated successfully with surgical excision of ulcer under amphotericin B/itraconazole coverage as primary cutaneous histoplasmosis confirmed pathologically and mycologically. A clinical suspicion remains paramount for early diagnosis of histoplasmosis particularly in a nonendemic area. Most importantly, with such diverse clinical presentation and therapeutic outcome selection of an appropriate and customized treatment schedule is a discretion the treating clinicians need to make.

Equine nasopharyngeal cryptococcoma due to Cryptococcus gattii

ABSTRACT: Cryptococcus gattii is often associated with pulmonary and systemic infections in humans and animals. In this research we report a case of nasopharyngeal cryptococoma caused by C. gatti in an equine. A 10-year-old mare presented a mass obstructing the oropharynx. Macroscopically the mass was asymmetric, and was attached to the ethmoidal sinuses and obstructed the oropharynx. Histopathological examination of the mass revealed multiple yeast cells ranging from spherical to oval, 4-8μm in diameter, with some of them showing narrow base polar budding. Cryptococcus gattii growth in mycological culture (Sabouraud Dextrose Agar) and was L-canavanine-glycine-bromothymol blue Agar positive. The molecular identification confirmed the isolate as C. gattii by means of the amplification of universal primers. C. gattii is considered an emerging fungal agent, as it affects human and animals and does not respond efficiently to commonly established treatments.

Onychomycosis InfectionsDo Polymerase Chain Reaction and Culture Reports Agree?

Mycological culture is the traditional method for identifying infecting agents of onychomycosis despite high false-negative results, slower processing, and complications surrounding nondermatophyte mold (NDM) infections. Molecular polymerase chain reaction (PCR) methods are faster and suited for ascertaining NDM infections. To measure agreement between culture and PCR methods for identification of infecting species of suspected onychomycosis, single toenail samples from 167 patients and repeated serial samples from 43 patients with suspected onychomycosis were processed by culture and PCR for identification of 16 dermatophytes and five NDMs. Agreement between methods was quantified using the kappa statistic (κ). The methods exhibited fair agreement for the identification of all infecting organisms (single samples: κ = 0.32; repeated samples: κ = 0.38). For dermatophytes, agreement was moderate (single samples: κ = 0.44; repeated samples: κ = 0.42). For NDMs, agreement was poor with single samples (κ = 0.16) but fair with repeated samples (κ = 0.25). Excluding false-negative reports from analyses improved agreement between methods in all cases except the identification of NDMs from single samples. Culture was three or four times more likely to report a false-negative result compared with PCR. The increased agreement between methods observed by excluding false-negative reports statistically clarifies and highlights the major discord caused by false-negative cultures. The increased agreement of NDM identification from poor to fair with repeated sampling along with their poor agreement in the single samples, with and without false-negatives, affirms the complications of NDM identification and supports the recommendation that serial samples help confirm the diagnosis of NDM infections.