Molecular characterization of clinical and environmental isolates from the Cryptococcus neoformans/C. Gattii species complexes of Maceió, Alagoas, Brazil.

Cryptococcosis is one of the major life-threatening opportunistic/systemic fungal diseases of worldwide occurrence, which can be asymptomatic or establish pneumonia and meningoencephalitis mainly in immunosuppressed patients, caused by the Cryptococcus neoformans and C. gattii species complexes. Acquisition is by inhaling fungal propagules from avian droppings, tree hollows and decaying wood, and the association of the molecular types with geographic origin, virulence and antifungal resistance have epidemiological importance. Since data on cryptococcosis in Alagoas are limited, we sought to determine the molecular types of etiological agents collected from clinical and environmental sources. We evaluated 21 isolates previously collected from cerebrospinal fluid and from environment sources (pigeon droppings and tree hollows) in Maceió-Alagoas (Brazil). Restriction fragment length polymorphism of URA5 gene was performed to characterize among the eight standard molecular types (VNI-VNIV and VGI-VGIV). Among isolates, 66.67% (14) were assigned to C. neoformans VNI - 12 of them (12/14) recovered from liquor and 2 from a tree hollow (2/14). One isolate from pigeon droppings (4.76%) corresponded to C. neoformans VNIV, while five strains from tree hollows and one from pigeon droppings (6, 28.57%) to C. gattii VGII. VNI-type was present in clinical and environmental samples and most C. neoformans infections were observed in HIV-positive patients, while types VNIV and VGII were prevalent in environmental sources in Alagoas. This is the first molecular characterization of Cryptococcus spp. in Alagoas, our study provides additional information on the ecoepidemiology of Cryptococcus spp. in Brazil, contributing to a closer view of the endemic species.

Molecular Detection of Medically Important Candida species from Droppings of Pigeons (Columbiformes) and Captive Birds (Passeriformes and Psittaciformes)

Abstract Passeriformes and Psittaciformes birds and pigeons (Columba livia) are known to be reservoirs of microorganisms, and their stool allows fungi development. Since accumulated avian excreta can interfere with public health, this study aimed to perform a molecular screening of medically important Candida species in pigeon droppings in public places and birds raised in captivity. Excreta collected from captive birds (3 residences) and pigeons (4 districts) were inoculated on Sabouraud dextrose agar with chloramphenicol for Gram staining and subculture on Hicrome® Candida. Three DNA extraction methods were performed for comparison (commercial kit, in-house and by boiling) and PCR to screen 6 clinically important Candida species among the isolates. The correlation between phenotypic and molecular methods was calculated by kappa/K. Only 6 C. parapsilosis (20%) were identified from captive birds' feces among 30 isolates (80% not identified), while pigeons' feces harbored a greater diversity, with the 6 pathogenic species confirmed among 41 isolates: C. albicans (31.70%/13), C. krusei (14.63%/6), C. tropicalis (14.63%/6), C. parapsilosis (17.10%/7), C. glabrata (14.63%/6) and C. guilliermondii (7.31%/3); 100% correlation between tested methods (K = 1) for the first 3 species. Boiling DNA extraction method was fast and efficient to obtain viable DNA from Candida spp. for PCR. Our results indicate that pigeon droppings harbor more potentially pathogenic species than birds in residential captivity, which probably have non-albicans Candida less frequently isolated in infectious processes. The greater availability of nutrients may have contributed to a diversity of Candida spp. in feces from public environments.

Molecular Identification of Candida Species in the Oral Microbiota of Individuals with Down Syndrome: A Case-Control Study.

Candida species are common in the human oral microbiota and may cause oral candidiasis (OC) when the microbiota equilibrium is disturbed. Immunosuppressed individuals are susceptible to oral infections as individuals with Down syndrome (IDS) due to particularities of their mouth morphoanatomy, saliva and comorbidities. This study aimed to analyze the molecular epidemiology of Candida spp. from the oral cavity of IDS and their relatives. A case-control study with 80 IDS and 80 non-syndromic (non-DS) was evaluated by oral swab collection for culture on Sabouraud dextrose agar, selection of yeast colonies to Gram staining and culturing on chromogenic media. DNA extraction was performed with the phenol/chloroform method for screening of 6 Candida species medically important in PCR, applying SPSS for statistics. We confirmed four species in 46 IDS without use of antimicrobials (57.54%), with a high prevalence of C. albicans/Ca (93.48%/43) and 3 C. glabrata/Cg (6.52%), being 25 of these IDS (31.25%) colonized by species other than C. albicans: Ca + Cg (16), Ca + C. tropicalis/Ct (7) and Ca + C. krusei/Ck (2). Only 10 non-DS were colonized by one species (11.25%): 6 C. albicans, 2 C. glabrata, 1 C. tropicalis and 1 C. krusei. Previous OC was reported by 39 IDS (48.75%), being 33 positives for Candida spp. (84.61%) and 17 with active OC (21.25%). Five non-DS reported OC previously and had no active lesions. Behavioral changes and buccal health programs directed to IDS may help prevent OC and its recurrence, providing information on oral hygiene for self-care.

Mycoses in northeastern Brazil: epidemiology and prevalence of fungal species in 8 years of retrospective analysis in Alagoas.

Superficial and cutaneous mycoses are common in tropical countries, caused by dermatophytes, yeast, and non-dermatophyte molds in different clinical specimens. In order to define the epidemiology of mycoses and the profile of their etiological agents in Alagoas (northeastern Brazil) between 2009 and 2016, we obtained data of patients from the main laboratories of Alagoas, by examining clinical samples with direct microscopy and culture on Sabouraud dextrose agar and Chromagar®Candida. A total of 3316 patients were confirmed with mycoses (595 men/2716 women) and 40.25 of average age. Positive samples totaled 3776, mainly vaginal secretion (1593/42.2%), toenails (876/23.2%), and fingernails (589/15.6%). Yeasts were the most isolated (3129/82.9%), including 3012 Candida spp. (79.8%), 57 Malassezia spp. (1.5%), 42 Trichosporon sp. (1.1%), 10 Geotrichum spp. (0.3%), and 8 Rhodotorula spp. (0.2%). Candida albicans was the most frequent species (715/18.9%), followed by C. krusei (194/5.1%), C. tropicalis (24/0.6%), and 2079 unspecified species (55.1%). Among 17.1% filamentous fungi, 14.8% dermatophytes were distributed as 211 Trichophyton sp. (5.6%), 125 T. rubrum (3.3%), 106 T. tonsurans (2.8%), 72 T. mentagrophytes (1.9%), 2 Microsporum sp. (0.1%), 15 M. canis (0.4%), and 26 Epidermophyton sp. (0.7%). Other fungi represented the minority: Fusarium sp. and Aspergillus sp. These are the first clinical data on the Alagoas population affected by fungi pathogens, confirming a higher incidence of candidiasis (mainly vulvovaginal and onychomycosis) and dermatophytes, providing a better understanding of different mycoses in northeastern Brazil.

Prevalence of ß-hemolytic Streptococcus in children with special health care needs.

UNLABELLED: Pharyngotonsillitis by ß-hemolytic Streptococcus mostly affects children and immunocompromised, being Streptococcus pyogenes (Group A) the most common agent in bacterial pharyngotonsillitis. AIM: This work targeted the research of ß-hemolytic Streptococcus Group-A (SBHGA) and No-A (SBHGNA) in the oropharynx of individuals with special health needs from the APAE (Maceió-AL). METHOD: A prospective study with oropharynx samples from patients with Down syndrome and other mental disorders (test) and students from a private school (control) aged 5-15 years. Cultures in blood agar (5%) were identified through Gram/catalase tests and bacitracin/trimethoprim-sulfamethoxazole disk diffusion method, applying the chi-squared statistical analysis. RESULTS: A total of 222 bacterial colonies were isolated in 74 individuals from APAE and 65 in the control group. In the test group, previous episodes of pharyngotonsillitis were reported by 36.49% (27/74) and 9.46% (7/74) were diagnosed with symptoms and/or signs suggestive of oropharynx infection. No positive sample of S. pyogenes was confirmed at APAE, being all samples classified as SBHGNA, with 5 SBHGA in the control group. CONCLUSION: The early identification of ß-hemolytic Streptococcus is important for the fast treatment of pharyngotonsillitis and the absence of S. pyogenes avoid future suppurative or not-suppurative sequels in the group from APAE.

Prevalência de Streptococcus β-hemolítico em crianças portadoras de necessidades especiais / Prevalence of β-hemolytic Streptococcus in children with special health care needs

Faringotonsilite causada por Streptococcus β-hemolítico afeta principalmente crianças e imunocomprometidos, sendo Streptococcus pyogenes (Grupo A) o agente mais comum em faringotonsilites bacterianas. OBJETIVO: Este trabalho objetivou a busca por Streptococcus β-hemolítico do Grupo A (SBHGA) e Não A (SBHGNA) na orofaringe de indivíduos com necessidades especiais da APAE (Maceió-AL). MÉTODO:Estudo prospectivo com amostras da orofaringe de pacientes com síndrome de Down e outras desordens mentais (teste) e estudantes de escola privada (controle) de 5-15 anos. Culturas em ágar sangue (5%) foram identificadas através dos testes de Gram/catalase e o método de disco difusão com bacitracina/sulfametoxazol-trimetoprim, aplicando-se o teste Chi-quadrado em análises estatísticas. RESULTADOS: Um total de 222 colônias bacterianas foram isoladas em 74 indivíduos da APAE e 65 no grupo controle. No grupo teste, episódios prévios de faringotonsilites foram relatados por 36,49% (27/74) e 9,46% (7/74) foram diagnosticados com sintomas e/ou sinais sugestivos de infecção orofaríngea. Nenhuma amostra de S. pyogenes foi confirmada na APAE, sendo todas identificadas como SBHGNA, com cinco SBHGA no grupo controle. CONCLUSÃO:A identificação precoce de Streptococcus β-hemolítico é importante para o tratamento rápido de faringotonsilites e a ausência de S. pyogenes evita futuras sequelas supurativas ou não supurativas no grupo da APAE.

Pharyngotonsillitis by β-hemolytic Streptococcus mostly affects children and imunocompromissed, being Streptococcuspyogenes (Group A) the most common agent in bacterial pharyngotonsillitis. AIM:This work targeted the research of β-hemolytic Streptococcus Group-A (SBHGA) and No-A (SBHGNA) in the oropharynx of individuals with special health needs from the APAE (Maceió-AL). METHOD: A prospective study with oropharynx samples from patients with Down syndrome and other mental disorders (test) and students from a private school (control) aged 5-15 years. Cultures in blood agar (5%) were identified through Gram/catalase tests and bacitracin/trimethoprim-sulfamethoxazole disk diffusion method, applying the chi-squared statistical analysis. RESULTS: A total of 222 bacterial colonies were isolated in 74 individuals from APAE and 65 in the control group. In the test group, previous episodes of pharyngotonsillitis were reported by 36.49% (27/74) and 9.46% (7/74) were diagnosed with symptoms and/or signs suggestive of oropharynx infection. No positive sample of S. pyogenes was confirmed at APAE, being all samples classified as SBHGNA, with 5 SBHGA in the control group. CONCLUSION: The early identification of β-hemolytic Steptococcus is important for the fast treatment of pharyngotonsillitis and the absence of S. pyogenes avoid future suppurative or not-suppurative sequels in the group from APAE.