Detecção de bactérias no ar em ambiente hospitalar com recurso a técnicas moleculares

A Qualidade do Ar Interior (QAI) revela-se de grande importância no ambiente hospitalar devido à disseminação aérea de bactérias potenciar as infecções nosocomiais. Os estudos nesta temática são raros em Portugal e inexistentes na Madeira. Este trabalho tem como objectivos identificar com elevada precisão a flora bacteriana aerosolizada no Hospital Dr. João de Almada (HJA) utilizando técnicas moleculares, determinar a sua origem, disseminação e especificidades e analisar globalmente a QAI. O ar exterior e de 15 locais no interior do HJA foi amostrado em Abril de 2009. Foram medidas a temperatura, humidade e concentração de CO2 e NO2 e quantificadas as bactérias e fungos no ar. Utilizaram-se testes bioquímicos e técnicas moleculares para caracterização e identificação de bactérias e foram comparadas comunidades bacterianas. A biodiversidade bacteriana no ar interior do HJA é dominada pelo género Staphylococcus (68%), sendo as espécies mais frequentes S. cohnii urealyticus, S. haemolyticus, S. capitis ou S. caprae e Micrococcus luteus. A maioria das espécies encontradas é comum em ambiente hospitalar e na flora comensal humana. As Staphylococcus spp. detectadas são consideradas patogénicas oportunistas envolvidas em infecções nosocomiais. A QAI é conforme com a legislação na maioria dos locais, havendo microorganismos em excesso em 12 das 72 amostras. O excesso de fungos relacionou-se com eventos de bruma no exterior e o excesso de bactérias correlacionou-se com a actividade humana e foi potenciado pela fraca ventilação e a presença ou manipulação de materiais contaminados. O ar exterior tem boa qualidade para ventilação do HJA, excepto durante eventos de bruma. As comunidades bacterianas aerosolizadas têm maior similaridade entre locais do mesmo tipo, sendo os ocupantes o principal foco de contaminação. São propostas medidas correctivas como o aumento da ventilação dos locais com elevada actividade humana e cuidados na manipulação de roupa suja e resíduos.

Avaliação do potencial de formigas (Hymenoptera: formicidae) como vetores mecânicos de bactérias do gêneroSstaphylococcus no ambiente hospitalar.

In a hospital environment, these bacteria can be spread by insects such as ants, which are characterized by high adaptability to the urban environment. Staphylococcus is a leading cause of hospital infection. In Europe, Latin America, USA and Canada, the group of coagulase negative staphylococci (CoNS) is the second leading cause of these infections, according to SENTRY (antimicrobial surveillance program- EUA). In this study, we investigated the potential of ants (Hymenoptera: Formicidae) as vehicle mechanics of Staphylococcus bacteria in a public hospital, in Natal-RN. The ants were collected, day and night, from June 2007 to may 2008, in the following sectors: hospitals, laundry, kitchen, blood bank. The ants were identified according to the identification key of Bolton, 1997. For the analysis of staphylococci, the ants were incubated in broth Tryptic Soy Broth (TSB) for 24 hours at 35 º C and then incubated on Mannitol Salt Agar. The typical colonies of staphylococci incubated for 24 hours at 35 ° C in Tryptic Soy Agar for the characterization tests (Gram stain, catalase, susceptibility to bacitracin and free coagulase). The identification of CoNS was performed through biochemical tests: susceptibility to novobiocin, growth under anaerobic conditions, presence of urease, the ornithine decarboxylation and acid production from the sugars mannose, maltose, trehalose, mannitol and xylose. The antimicrobial susceptibility examined by disk-diffusion technique. The technique of Polymerase Chain Reaction was used to confirm the presence of mecA gene and the ability to produce biofilm was verified by testing in vitro using polystyrene inert surface, in samples of resistant staphylococci. Among 440 ants, 85 (19.1%) were carrying coagulase-negative staphylococci (CoNS) of the species Staphylococcus saprophyticus (17), Staphylococcus epidermidis (15), Staphylococcus xylosus (13), Staphylococcus hominis hominis (10), Staphylococcus lugdunensis (10), Staphylococcus warneri (6), Staphylococcus cohnii urealyticum (5), Staphylococcus haemolyticus (3), Staphylococcus simulans (3), Staphylococcus cohnii cohnii (2), and Staphylococcus capitis (1). No Staphylococcus aureus was found. Among the isolates, 30.58% showed resistance to erythromycin. Two samples of CoNS (2.35%), obtained from the ant Tapinoma melanocephalum collected in the post-surgical female ward, S. Hominis hominis and S. lugdunensis harbored the mecA gene and were resistant to multiple antibiotics, and the specie S. hominis hominis even showed to be a biofilm producer. This study proves that ants act as carriers of multidrug-resistant coagulase-negative Staphylococci and biofilm producers and points to the risk of the spreading of pathogenic microorganisms by this insect in the hospital environment

Contribuição ao estudo da reação á tricofitina nas dermatofitoses

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Microsporum nanum. quarto relato de infecção humana no Brasil

Comunica-se o quarto relato de infecção humana por Microsporum nanum (M. nanun) no Brasil. Trata-se de criança de 9 meses de idade, apresentando há um mês lesões cutâneas dorsais compatíveis com tinea corporis. As lesões foram frustas, regredindo espontaneamente após um mês. A investigação epidemiológica na área de origem do caso índice identificou suínos infectados por Microsporum nanum, sugerindo ser esta a fonte de infecção.

Polymerase Chain Reaction Detection of Enterotoxins Genes in Coagulase-Negative Staphylococci Isolated from Brazilian Minas Cheese

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cladophialophora saturnica sp nov., a new opportunistic species of Chaetothyriales revealed using molecular data

While many members of the black yeasts genus Cladophialophora have been reported to cause diseases in humans, understanding of their natural niche is frequently lacking. Some species can be recovered from the natural environment by means of selective isolation techniques. The present study focuses on a Cladophialophora strain that caused an interdigital tinea nigra-like lesion in a HIV-positive Brazilian child. The fungal infection was successfully treated with oxiconazole. Similar strains had been recovered from the environment in Brazil, Uruguay and the Netherlands. The strains were characterized by sequencing the Internal Transcribed Spacer (ITS) regions and the small subunit (SSU) of the nuclear ribosomal RNA gene, as well as the elongation factor 1-alpha (EF1) gene. Since no match with any known species was found, it is described as the new species, Cladophialophora saturnica.

Effect of photodynamic therapy on clinical isolates of Staphylococcus spp

Staphylococcus spp. are opportunistic microorganisms known for their capacity to develop resistance against antimicrobial agents. The objective of this study was to evaluate the effect of photodynamic therapy (PDT) on 20 Staphylococcus strains isolated from the human oral cavity, including S. aureus, S. schleiferi, S. epidermidis, S. capitis, S. haemolyticus, and S. lentus. A suspension of each Staphylococcus strain (10(6) cells/mL) was submitted to PDT using methylene blue and a low power laser. The isolated effects of methylene blue, laser treatment and ciprofloxacin were also evaluated. After the experimental treatments, 0.1 mL aliquots of the suspensions were seeded onto BHI agar for determination of the number of colony-forming units (CFU/mL). The results were analyzed by analysis of variance and Tukey's test (p < 0.05). The mean reduction in bacterial counts of the strains submitted to PDT ranged from 4.89 to 6.83 CFU (log10)/mL, with the observation of a decreasing susceptibility to treatment of S. schleiferi, S. haemolyticus, S. epidermidis, S. capitis, S. aureus, and S. lentus. The results showed that PDT was effective in reducing the number of viable cells of all clinical Staphylococcus isolates studied.

Avaliação da colonização nasal por Staphylococcus spp. resistenteà oxacilina em alunos de enfermagem

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Modelo de carroceria e seu impacto sobre o bem-estar e a qualidade da carne dos suínos

O objetivo deste trabalho foi avaliar o efeito do modelo de carroceria (MC) (metálica dupla ou simples e simples de madeira) sobre a freqüência de diferentes tipos de lesões na carcaça (FLC) e sobre a qualidade da carne dos suínos. Foram utilizadas 360 fêmeas com peso vivo médio de 132,72±11,09kg. Para a análise estatística, considerou-se no modelo os efeitos de bloco (BL), o MC e a interação entre MC x BL. Verificou-se um efeito do MC sobre a freqüência de lesões na carcaça provenientes de briga e a freqüência total de lesões, sobre o pH1 do músculo Semispinalis capitis (SC), sobre o pH U dos músculos SC, Longissimus dorsi (LD) e Semimembranosus (SM) e sobre a cor dos músculos LD e SM. Suínos transportados em carroceria metálica simples apresentaram maiores valores de pH e da cor dos músculos avaliados. Conclui-se que suínos transportados em carroceria simples apresentam maior prevalência de lesões na carcaça e que o transporte dos suínos em carroceria metálica teve efeito positivo sobre os valores de pH e da cor carne.

Microsporum nanum relato do segundo caso de infecção humana no Brasil

É comunicado o segundo caso de infecção humana por Microsporum nanum no Brasil. A investigação epidemiológica visando determinar a fonte de infecção não obteve sucesso. O padrão dermatológico das lesões foi o de Tinea corporis clássica.

Efeito das condições pré-abate sobre a qualidade da carne de suínos pesados

The objective of this study was to evaluate pig fasting time at the farm (FT= 9, 12, 15 or 18 hours) and pig position into the pig lorry compartment on pork quality through liquid drip loss (DLL), pH1 evaluated at 45 minutes post slaughter and pHu evaluated 24 hours post slaughter. One hundred ninety two females, weighing 133±11 kg, from two farms were used. Pig locations were evaluated on truck compartment considering front, middle and rear (TCL) position and top or botton decks (LUD). The following effects were considered in the statistical model: block (BL= farm), FT, TCL, LUD and the interaction between BL and FT. The FT affected significantly the pH1 of Longissimus dorsi (LD) muscle, and pHu of Semispinalis capitis (SC), Longissimus dorsi (LD) and Semimembranosus (SM) muscles. Fasting time at farm equal or shorter than 12 hours resulted in carcasses with lower pHu values at LD and SM muscles and fasting time above 15 hours resulted in higher pHu on SC. There were no observed effect (p>0.05) of the evaluated sources of variation on DLL. TCL affected pH1 of SC, LD and SM muscles, and pHu of LD and SM muscles. Pigs located in middle or rear position in the lorry had no difference in pH1 of the evaluated muscles. But pigs located in the middle position of the lorry had greater values of pHu on LD and SM. It is stated that pigs fasting time at farm need to be close to 15 hours in aim of obtain higher frequency of pHu values in the normal range (5.55

Enterotoxin genes in coagulase-negative and coagulase-positive staphylococci isolated from bovine milk

The objective of this study was to isolate and identify the main staphylococcal species causing bovine mastitis in 10 Brazilian dairy herds and study their capability to produce enterotoxins. Herds were selected based on size and use of milking technology, and farms were visited once during the study. All mammary glands of all lactating cows were screened using the California Mastitis Test (CMT) and a strip cup. A single aseptic milk sample (20. mL) was collected from all CMT-positive quarters. Identification of Staphylococcus spp. was performed using conventional microbiology, and PCR was used to determine the presence of enterotoxin-encoding genes (sea, seb, sec, and sed). Of the 1,318 CMT-positive milk samples, Staphylococcus spp. were isolated from 263 (19.9%). Of these isolates, 135 (51%) were coagulase-positive staphylococci (CPS) and 128 (49%) were coagulase-negative staphylococci (CNS). Eighteen different species of CNS were isolated, among which S. warneri, S. epidermidis and S. hyicus were the most frequent. The distribution of Staphylococcus species was different among herds: S. epidermidis was found in 8 herds, S. warneri was found in 7 herds, and S. hyicus in 6 herds. Some of the CNS species (S. saprophyticus ssp. saprophyticus, S. auricularis, S. capitis, and S. chromogenes) were isolated in only one of the farms. Genes related to production of enterotoxins were found in 66% (n = 85) of all CNS and in 35% of the CPS isolates. For both CNS and CPS isolates, the most frequently identified enterotoxin genes were sea, seb, and sec; the prevalence of sea differed between CPS (9.5%) and CNS (35.1%) isolates. Staphylococcus warneri isolates showed a greater percentage of sea than seb, sec, or sed, whereas S. hyicus isolates showed a greater percentage of sea than sec. Over 60% of CNS belonged to 3 major species, which carried 62.2 to 81.3% of the enterotoxin genes. The high prevalence highlights the potential for food poisoning caused by these species. For possible high-risk situations for food poisoning, such as milk produced with total bacterial counts greater than regulatory levels and stored under inappropriate temperatures, monitoring contamination with CNS could be important to protect human health. Because the prevalence of CNS intramammary infections in dairy herds is usually high, and these species can be found in great numbers in bulk milk, identification of risk factors for production of staphylococcal enterotoxins should be considered in future studies. © 2013 American Dairy Science Association.

Bidestilação em alambiques contendo dispositivos de prata e cobre e sua influência na qualidade da cachaça

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Enterotoxinas de Staphylococus coagulase positiva e negativa isoladas das fossas nasais e mãos de manipuladores de alimentos

Pós-graduação em Medicina Veterinária - FMVZ

Perfil de sensibilidade microbiana, pesquisa de gene mecA de resistência à meticilina e detecção molecular de genes codificadores de enterotoxinas, em espécies de estafilococos coagulase positiva e negativa, isolados de mastites bovinas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)