Clinical and antimicrobial efficacy of NitrAdineTM-based disinfecting cleaning tablets in complete denture wearers

OBJECTIVE: This study evaluated the efficacy of NitrAdineTM-based disinfecting cleaning tablets for complete denture, in terms of denture biofilm removal and antimicrobial action. MATERIAL AND METHODS: Forty complete denture wearers (14 men and 26 women) with a mean age of 62.3±9.0 years were randomly assigned to two groups and were instructed to clean their dentures according to two methods: brushing (control) - 3 times a day with denture brush and tap water following meals; brushing and immersion (Experimental) - brushing the denture 3 times a day with denture brush and tap water following meals and immersion of the denture in NitrAdineTM-based denture tablets (Medical InterporousTM). Each method was used for 21 days. Denture biofilm was disclosed by a 1% neutral red solution and quantified by means of digital photos taken from the internal surface before and after the use of the product. Microbiological assessment was conducted to quantify Candida sp. RESULTS: An independent t-test revealed a significant lower biofilm percentage for the experimental group (4.7, 95% CI 2.4 to 7.9) in comparison with the control group (mean 37.5, 95% CI 28.2 to 48.1) (t38=7.996, p<0.001). A significant reduction of yeast colony forming units could be found after treatment with Medical InterporousTM denture tablets as compared to the control group (Mann-Whitney test, Z=1.90; p<0.05). CONCLUSION: The present findings suggest that NitrAdineTM-based disinfecting cleaning tablets are efficient in removal of denture biofilm. In addition, a clear antimicrobial action was demonstrated. Therefore, they should be recommended as a routine denture maintenance method for the prevention of the development of microbial biofilm induced denture stomatitis.

Antimicrobial efficacy of chemical disinfectants on contaminated full metal crowns

Prosthetic restorations that have been tried in the patient's mouth are potential sources of infection. In order to avoid cross-infection, protocols for infection control should be established in dental office and laboratory. This study evaluated the antimicrobial efficacy of disinfectants on full metal crowns contaminated with microorganisms. Full crowns cast in a Ni-Cr alloy were assigned to one control group (n=6) and 5 experimental groups (n=18). The crowns were placed in flat-bottom glass balloons and were autoclaved. A microbial suspension of each type of strain - S. aureus, P. aeruginosa, S. mutans, E. faecalis and C. albicans- was aseptically added to each experimental group, the crowns being allowed for contamination during 30 min. The contaminated specimens were placed into recipients with the chemical disinfectants (1% and 2% sodium hypochlorite and 2% glutaraldehyde) for 5, 10 and 15 min. Thereafter, the crowns were placed into tubes containing different broths and incubated at 35ºC. The control specimens were contaminated, immersed in distilled water for 20 min and cultured in Thioglycollate broth at 35ºC. Microbial growth assay was performed by qualitative visual examination after 48 h, 7 and 12 days. Microbial growth was noticed only in the control group. In the experimental groups, turbidity of the broths was not observed, regardless of the strains and immersion intervals, thus indicating absence of microbial growth. In conclusion, all chemical disinfectants were effective in preventing microbial growth onto full metal crowns.

Isolation of Pseudomonas aeruginosa strains from dental office environments and units in Barretos, state of São Paulo, Brazil, and analysis of their susceptibility to antimicrobial drugs

A wide variety of opportunistic pathogens has been detected in the tubing supplying water to odontological equipment, in special in the biofilm lining of these tubes. Among these pathogens, Pseudomonas aeruginosa, one of the leading causes of nosocomial infections, is frequently found in water lines supplying dental units. In the present work, 160 samples of water, and 200 fomite samples from forty dental units were collected in the city of Barretos, State of São Paulo, Brazil and evaluated between January and July, 2005. Seventy-six P. aeruginosa strains, isolated from the dental environment (5 strains) and water system (71 strains), were tested for susceptibility to six antimicrobial drugs most frequently used against P. aeruginosa infections. Susceptibility to ciprofloxacin, followed by meropenem was the predominant profile. The need for effective means of reducing the microbial burden within dental unit water lines is emphasized, and the risk of exposure and cross-infection in dental practice, in special when caused by opportunistic pathogens like P. aeruginosa, are highlighted.

Maximum inhibitory dilution of mouthwashes containing chlorhexidine and polyhexamethylene biguanide against salivary staphylococcus aureus

OBJECTIVE: The aim of the present study was to determine the in vitro maximum inhibitory dilution (MID) of two chlorhexidinebased oral mouthwashes (CHX): Noplak®, Periogard®, and one polyhexamethylene biguanide-based mouthwash (PHMB): Sanifill Premium® against 28 field Staphylococcus aureus strains using the agar dilution method. MATERIALS AND METHODS: For each product, decimal dilutions ranging from 1/10 to 1/655,360 were prepared in distilled water and added to Mueller Hinton Agar culture medium. After homogenization, the culture medium was poured onto Petri dishes. Strains were inoculated using a Steers multipoint inoculator and dishes were incubated at 37ºC for 24hours. For reading, MID was considered as the maximum dilution of the mouthwash still capable of inhibiting microbial growth. RESULTS: Sanifill Premium® inhibited the growth of all strains at 1/40 dilution and of 1 strain at 1/80 dilution. Noplak® inhibited the growth of 23 strains at 1/640 dilution and of all 28 strains at 1/320 dilution. Periogard® showed inhibited growth of 7 strains at 1/640 dilution and of all 28 strains at 1/320 dilution. Data were submitted to Kruskal-Wallis statistical test, showing significant differences between the mouthwashes evaluated (p<0.05). No significant difference was found between Noplak® and Periogard® (p>0.05). Sanifill Premium® was the least effective (p<0.05). CONCLUSION: It was concluded that CHX-based mouthwashes present better antimicrobial activity against S. Aureus than the PHMB-based mouthwash.

Determination of the maximum inhibitory dilution of cetylpyridinium chloride-based mouthwashes against staphylococcus aureus: an in vitro study

The aim of this in vitro study was to determine the maximum inhibitory dilution (MID) of four cetylpyridinium chloride (CPC)-based mouthwashes: CPC+Propolis, CPC+Malva, CPC+Eucaliptol+Juá+Romã+Propolis (Natural Honey®) and CPC (Cepacol®), against 28 Staphylococcus aureus field strains, using the agar dilution method. Decimal dilutions ranging from 1/10 to 1/655,360 were prepared and added to Mueller Hinton Agar. Strains were inoculated using Steers multipoint inoculator. The inocula were seeded onto the surface of the culture medium in Petri dishes containing different dilutions of the mouthwashes. The dishes were incubated at 37ºC for 24 h. For readings, the MID was considered as the maximum dilution of mouthwash still capable of inhibiting microbial growth. The obtained data showed that CPC+Propolis had antimicrobial activity against 27 strains at 1/320 dilution and against all 28 strains at 1/160 dilution, CPC+Malva inhibited the growth of all 28 strains at 1/320 dilution, CPC+Eucaliptol+Juá+Romã+Propolis inhibited the growth of 2 strains at 1/640 dilution and all 28 strains at 1/320 dilution, and Cepacol® showed antimicrobial activity against 3 strains at 1/320 dilution and against all 28 strains at 1/160 dilution. Data were submitted to Kruskal-Wallis test, showing that the MID of Cepacol® was lower than that determined for the other products (p<0.05). In conclusion, CPC-mouthwashes showed antimicrobial activity against S. aureus and the addition of other substances to CPC improved its antimicrobial effect.

Carcass quality of feedlot finished steers fed yeast, monensin, and the association of both additives

To evaluate the effects of the supplementation of feed additives on carcass quality in beef cattle, 72 Nellore steers (339.5kg, 20-month old) were feedlot finished and fed for 91 days one of the following diets: 1) control with no additives; or added of 2) live yeast culture; 3) monensin; or 4) the association of both additives. After slaughter, renal, pelvic, and inguinal fat and hot carcass weights were recorded and carcass was split into muscle, bone, and trimmable fat. Carcass Longissimus muscle area and subcutaneous fat thickness at the 12th rib were measured and steaks of Longisimus muscle were taken to determine meat color, shear force, drip, and cooking losses. Yeast increased carcass dressing percentage but there were no effects on hot carcass weight, Longissimus area, subcutaneous fat thickness, percentage and weight of retail cut yield and trimmings. Feed additives had no effect on carcass pH, meat color, fat content, shear force, and drip losses. Supplementation of yeast, monensin or the association of both additives had no important effects on carcass traits and on meat quality of feedlot finished steers.

Aves da Estação Ecológica de Itirapina, estado de São Paulo, Brasil

O cerrado é um dos biomas mais ameaçados no mundo, apresentando algumas das maiores taxas de destruição causadas principalmente pela agropecuária. Poucos estudos têm avaliado a diversidade local de aves no cerrado, especialmente em áreas marginais desse bioma. A Estação Ecológica de Itirapina (EEI) contém um dos últimos remanescentes de campos naturais e cerrados do estado de São Paulo. Um levantamento das aves desta unidade de conservação, feito tanto por observações casuais como sistemáticas entre 1998 e 2007, e complementado por revisão de literatura, revelou que 231 espécies apresentaram ocorrência recente na área. Desse total, 38 espécies (16,4%) encontram-se na lista de espécies ameaçadas para o estado de São Paulo e/ou são endêmicas do Cerrado, um alto número quando comparado com outras áreas de cerrados paulistas ou mesmo em relação ao Brasil Central. Tal fato reforça a importância da EEI em relação à conservação da avifauna e demonstra que a mesma está mantendo sua função de conservar a biodiversidade. Adicionalmente, ao longo dos 10 anos de estudo foram detectadas várias ameaças à EEI, e neste sentido são recomendadas aqui algumas sugestões de manejo que consideramos essenciais para essa unidade de conservação manter ou mesmo aumentar sua diversidade de aves.

Estádio sucessional e fatores geográficos como determinantes da similaridade florística entre comunidades florestais no Planalto Atlântico, Estado de São Paulo, Brasil

A análise das relações de similaridade florística entre comunidades geralmente conduz ao estabelecimento de padrões, condicionados por fatores diversos que determinam a ocorrência ou não das espécies em diferentes locais. Em busca de tais padrões, foram analisadas as relações de similaridade florística entre comunidades florestais localizadas na região do Planalto de Ibiúna, estado de São Paulo, Brasil. Incluíram-se na análise 21 fragmentos florestais e seis sítios em uma Reserva Florestal contínua, sendo que a composição florística e a estrutura da comunidade arbórea (DAP mínimo 5 cm) em cada local foram amostradas pelo método de quadrantes. Aplicaram-se dois métodos de análises multivariadas: 1) Análise de Correspondência Destendenciada (DCA), com base no índice de similaridade de Sørensen; e 2) Divisão Hierárquica Dicotômica (TWINSPAN). A similaridade florística foi mais elevada entre comunidades em estádios sucessionais semelhantes, especialmente se estivessem geograficamente próximas. Há um gradiente florístico associado à latitude, indicando tratar-se de uma região de transição entre biomas. Nos sítios situados na face norte da região de estudo estão presentes espécies que também ocorrem no cerradão e em floresta estacional semidecidual, enquanto nos sítios situados na face sul prevalecem espécies características da floresta ombrófila densa.

Screening for antifungal activities of extracts of the brazilian seaweed genus Laurencia (Ceramiales, Rhodophyta)

The resistance of pathogens to commonly used antibiotics has enhanced morbidity and mortality and has triggered the search for new drugs. Several species of the red alga genus Laurencia are very interesting candidates as potential sources of natural products with pharmaceutical activity because they are known to produce a wide range of chemically interesting halogenated secondary metabolites. This is an initial report of the antifungal activities of the secondary metabolites of five species of Laurencia, collected in the state of Espírito Santo, against three strains of pathogenic fungi: Candida albicans (CA), Candida parapsilosis (CP), and Cryptococcus neoformans (CN). Minimum inhibitory concentrations (MIC) of the algal extracts were determined by serial dilution method in RPMI 1640 Medium in 96-well plates according to the NCCLS and microbial growth was determined by absorbance at 492nm. A result showing maintenance or reduction of the inoculum was defined as fungistatic, while fungicidal action was no observed growth in the 10 µL fungistatic samples subcultured in Sabouraud Agar. Our results indicate that apolar extracts of Laurencia species possess antifungal properties and encourage continued research to find new drugs for therapy of infectious diseases in these algae.

Antibiosis and dark-pigments secretion by the phytopathogenic and environmental fungal species after interaction in vitro with a Bacillus subtilis isolate

In this work, different reactions in vitro between an environmental bacterial isolate and fungal species were related. The Gram-positive bacteria had terminal and subterminal endospores, presented metabolic characteristics of mesophilic and acidophilic growth, halotolerance, positive to nitrate reduction and enzyme production, as caseinase and catalase. The analysis of partial sequences containing 400 to 700 bases of the 16S ribosomal RNA gene showed identity with the genus Bacillus. However, its identity as B. subtilis was confirmed after analyses of the rpoB, gyrA, and 16S rRNA near-full-length sequences. Strong inhibitory activity of environmental microorganisms, such as Penicillium sp, Aspergillus flavus, A. niger, and phytopathogens, such as Colletotrichum sp, Alternaria alternata, Fusarium solani and F. oxysporum f.sp vasinfectum, was shown on co-cultures with B. subtilis strain, particularly on Sabouraud dextrose agar (SDA) and DNase media. Red and red-ochre color pigments, probably phaeomelanins, were secreted by A. alternata and A. niger respectively after seven days of co-culture.

Geographical and seasonal distributions of the seedeaters Sporophila bouvreuil and Sporophila pileata (aves: Emberizidae)

Many species in the genus Sporophila are migratory. Migration patterns, while poorly studied, may be influenced by seed production which can be very seasonal in some regions. The distribution of S. bouvreuil extends from the Amazon and Suriname south through a large part of the open regions of Brazil. Sporophila pileata, on the other hand, is found in southeastern and southern Brazil as well as Argentina and Paraguay. Both of these species migrate, but their movement patterns are poorly known. To better understand the geographical and the seasonal distributions of S. bouvreuil and S. pileata, we grouped the records into two categories: the breeding season (September to March) and the putative migration season (April to August). We found two areas of sympatry between S. bouvreuil and S. pileata in the Brazilian states of Minas Gerais and São Paulo. For S. bouvreuil we suggest that populations that breed in the Amazon migrate to the Cerrado or Caatinga, where they will encounter resident populations of the same species. These resident populations may take part in short distance migrations. Sporophila pileata, on the other hand, occur in the Cerrado and open areas within the Atlantic Forest and it is not yet possible to determine migratory tendencies or destinations in the non-breeding season.

Butterflies (Lepidoptera, Papilionoidea and Hesperioidea) of the "Baixada Santista" region, coastal São Paulo, southeastern Brazil

Butterflies (Lepidoptera, Papilionoidea and Hesperioidea) of the "Baixada Santista" region, coastal São Paulo, southeastern Brazil. A list with 538 species of butterflies recorded in the Baixada Santista, São Paulo ( SE Brazil) is presented. Standard sampling protocols (i.e. with entomological nets) were followed. Baited traps were installed for fruit feeding species. Data from the literature and entomological collections were also considered in the total estimated species richness. The species richness recorded in the Baixada Santista region represents about 16% of the Brazilian butterfly fauna, and 34% of the known butterfly fauna for the state of São Paulo. The present list contains an appreciably higher number of species in comparison to other lists from similar biomes farther south, such as Blumenau in Santa Catarina, and Maquiné in Rio Grande do Sul.

Do Código Florestal para o Código da Biodiversidade

Em face do gigantismo do território e da situação real em que se encontram os seus macro biomas - Amazônia Brasileira, Brasil Tropical Atlântico, Cerrados do Brasil Central, Planalto das Araucárias, e Pradarias Mistas do Brasil Subtropical - e de seus numerosos mini-biomas, faixas de transição e relictos de ecossistemas, qualquer tentativa de mudança no "Código Florestal" tem que ser conduzido por pessoas competentes e bioeticamente sensíveis. Por muitas razões, se houvesse um movimento para aprimorar o atual Código Florestal, teria que envolver o sentido mais amplo de um Código de Biodiversidades, levando em conta o complexo mosaico vegetacional de nosso território. Enquanto o mundo inteiro trabalha para a diminuição radical de emissão de CO2, o projeto de reforma proposto na Câmara Federal de revisão do Código Florestal defende um processo que significará uma onda de desmatamento e emissões incontroláveis de gás carbônico, fato observado por muitos críticos em diversos trabalhos e entrevistas. A utopia de um desenvolvimento com o máximo de florestas em pé não pode ser eliminada por princípio em função de mudanças radicais do Código Florestal, sendo necessário pensar no território total de nosso país, sob um ampliado e correto Código de Biodiversidade. Ou seja, um pensamento que envolva: as nossas grandes florestas (Amazônia e Matas Tropicais Atlânticas); o domínio das caatingas e agrestes sertanejos; planaltos centrais com cerrados, cerradões e campestres; os planaltos de araucárias sul-brasileiros; as pradarias mistas do Rio Grande do Sul; e os redutos e mini-biomas da costa brasileira e do Pantanal mato-grossense, e faixas de transição e contrato (core-áreas) de todos os domínios morfoclimáticos e fitogeográficos brasileiros).

Heterologous expression of glucose oxidase in the yeast Kluyveromyces marxianus

Background: In spite of its advantageous physiological properties for bioprocess applications, the use of the yeast Kluyveromyces marxianus as a host for heterologous protein production has been very limited, in constrast to its close relative Kluyveromyces lactis. In the present work, the model protein glucose oxidase (GOX) from Aspergillus niger was cloned into K. marxianus CBS 6556 and into K. lactis CBS 2359 using three different expression systems. We aimed at verifying how each expression system would affect protein expression, secretion/localization, post-translational modification, and biochemical properties. Results: The highest GOX expression levels (1552 units of secreted protein per gram dry cell weight) were achieved using an episomal system, in which the INU1 promoter and terminator were used to drive heterologous gene expression, together with the INU1 prepro sequence, which was employed to drive secretion of the enzyme. In all cases, GOX was mainly secreted, remaining either in the periplasmic space or in the culture supernatant. Whereas the use of genetic elements from Saccharomyces cerevisiae to drive heterologous protein expression led to higher expression levels in K. lactis than in K. marxianus, the use of INU1 genetic elements clearly led to the opposite result. The biochemical characterization of GOX confirmed the correct expression of the protein and showed that K. marxianus has a tendency to hyperglycosylate the protein, in a similar way as already observed for other yeasts, although this tendency seems to be smaller than the one of e. g. K. lactis and S. cerevisiae. Hyperglycosylation of GOX does not seem to affect its affinity for the substrate, nor its activity. Conclusions: Taken together, our results indicate that K. marxianus is indeed a good host for the expression of heterologous proteins, not only for its physiological properties, but also because it correctly secretes and folds these proteins.

Screening of Filamentous Fungi to Identify Biocatalysts for Lupeol Biotransformation

The goal of the study was to evaluate the ability of filamentous fungi to biotransform the pentacyclic triterpene lupeol. The microbial transformations were carried out in shake flasks in different media. Experiments were also run with control flasks. Samples of each culture were taken every 24 hours, extracted with ethyl acetate, and analyzed by GC-MS. The biotransformation of lupeol by Aspergillus ochraceus and Mucor rouxii afforded two compounds in each culture, which were detected in the cultures developed for more than seven days only in the Koch's K1 medium. The obtained data demonstrated that A. ochraceus is a good biocatalyst to introduce double bonds in the lupeol structure, whereas M. rouxii exhibits ability to biocatalyze oxygen insertions in that pentacyclic triterpene. Mass spectrometry was demonstrated to be an efficient analytical method to select promising biocatalysts for the compound investigated in this study. The biotransformation processes were influenced by the culture medium and incubation period. The obtained results open the perspective of using A. ochraceus and M. rouxii in pentacyclic triterpene biotransformations.