Bacterial vaginosis biofilms: challenges to current therapies and emerging solutions

Bacterial vaginosis (BV) is the most common genital tract infection in women during their reproductive years and it has been associated with serious health complications, such as preterm delivery and acquisition or transmission of several sexually transmitted agents. BV is characterized by a reduction of beneficial lactobacilli and a significant increase in number of anaerobic bacteria, including Gardnerella vaginalis, Atopobium vaginae, Mobiluncus spp., Bacteroides spp. and Prevotella spp.. Being polymicrobial in nature, BV etiology remains unclear. However, it is certain that BV involves the presence of a thick vaginal multi-species biofilm, where G. vaginalis is the predominant species. Similar to what happens in many other biofilm-related infections, standard antibiotics, like metronidazole, are unable to fully eradicate the vaginal biofilm, which can explain the high recurrence rates of BV. Furthermore, antibiotic therapy can also cause a negative impact on the healthy vaginal microflora. These issues sparked the interest in developing alternative therapeutic strategies. This review provides a quick synopsis of the currently approved and available antibiotics for BV treatment while presenting an overview of novel strategies that are being explored for the treatment of this disorder, with special focus on natural compounds that are able to overcome biofilm-associated antibiotic resistance.

The relevance of bacterial interactions on oral health

Dissertação de mestrado integrado em Engenharia Biomédica (área de especialização em Engenharia Clínica)

Role of biofilms in the oral health of animals

In humans the importance of biofilms in disease processes is now widely recognised together with the difficulties in treating such infections once established. One of the earliest and certainly most studied biofilm in humans is that of dental plaque which is responsible for two of the most prevalent human infections, namely dental caries and periodontal disease. However, comparable studies of dental plaque in animals are relatively limited, despite the fact that similar infections also occur, and in the case of farm animals there is an associated economic impact. In addition, biofilms in the mouths of animals can also be detrimental to human health when transferred by animal bites. As a result, an understanding of both the microbial composition of animal plaque biofilms together with their role in animal diseases is important. Through the use of modern molecular studies, an insight into the oral microflora of animals is now being obtained and, to date, reveals that despite differences in terms of microbial species and relative proportions occurring between humans and animals, similarities do indeed exist. This information can be exploited in our efforts to both manage and treat infections in animals arising from the presence of an oral biofilm. This Chapter describes our current understanding of the microbial composition of animal plaque, its role in disease and how oral hygiene measures can be implemented to reduce subsequent infection.

Avaliação da capacidade de fixação de fósforo pelo solo

Phosphate "fixation" is the convertion of soluble into insoluble phosphate in the soil. There are many factors conditioning phosphate fixation by soil such as reactions originating less soluble compounds (phosphates of iron, aluminum, calcium, magnesium, etc.), PO4-3 adsorption by the colloidal fraction of soils, PO4-3 absorption by the soil microflora, etc. Certain soils of the state of São Paulo (Brazil) are relatively rich in both iron and aluminum oxides. PO4-3 fixation, using P31 and P32 has been verified by researchers, specially with "Terra Roxa". The known methods for fixation evaluation are conventional as this depends on phosphate solution concentration, pH, time of contact between soil and solution, relation of sample weight to solution volume, shaking time, etc. In this experiment, the following conventional method was used: 4 g of soil were shaken for 15 minutes at 30-40 rpm, in 300 ml Erlenmeyer flask in a Wagner shaking machine, together with 100 ml of 0,03 normal phosphate solution (being 0,01 normal as PO4-3 contributed by H8PO4 and 0,02 normal as PO4-3 from KH2PO4). After shaking it was set aside for 24 hours and then filtered. Phosphate was determined in a suitable aliquot of both the original solution (blank) and the soil extract, by the vanadomolibidic-phosphoric acid method. From phosphate concentration in the blank minus phosphate concentration in the soil stract the rate of fixation by 100 g of soil was calculated. The data obtained show that "Terra Roxa" and "Terra Roxa Misturada" have a fairly high PO4-3 fixation capacity, varying from 10 to 24 milliequivalents of PO4-3 per 100 g of soil.

Ocorrência de Actinomicetos com atividade proteolitica, em um solo cultivado

Among 327 strains of Actinomycetes isolated in 1958 during the four seasons, 97,85 per cent had proteolytic activity. Most of the strains wete isolated in spring time. About the organic matter no conclusions were reached, because it was mantained constant with a average value of 4.099. So the different number of the strains was a consequence of the season. We can conclude that the Latosol red-yellow soil, from where these microorganisms were isolated, has a strong proteolytic capacity, since this reaction is very strong in a maximum limit of sixteen days during the spring season. Consequently, this soil has one of the most importante caracteristics of fertility. The role of the Actinomycetes in the soil is so important of the soil microflora.

Atividade proteolítica e amonificante de vários tipos de solo do estado de São Paulo. I. - Solo latosol vermelho amarelo, com e sem cobertura morta

Os autores estudam, no presente trabalho, um solo tipo Latosol Vermelho Amarelo, com e sem cobertura morta, em 2 profundidades diferentes. Concluiram sôbre as vantagens de se manter um solo com cobertura morta que permite um aumento da microflora total e dos elementos nutritivos, confirmado pelo pronunciado índice de atividade protelítica e amonificante do solo.

Técnica do xenodiagnostico na molestia de Chagas

O xenodiagnostico é um processo muito valioso para o diagnostico etiologico da moléstia de Chagas. Consiste em alimentar-se barbeiros normais em supótos portadores da infecção e determinar-se, depois, si eles adquirem ou não o parasitismo. Em cada região deve ser empregada, de preferência, o transmissor local mais importante. Três a seis ninfas famintas são geralmente empregadas em cada prova, devendo ficar em contacto com o doador até encherem-se completamente (15-30 minutos). Si o exame de fezes – espontaneamente eliminadas ou obtidas por punção anal – não revelar a presença de flagelados, os insetos devem ser dissecados 40 a 60 dias depois da sucção, para exame do conteúdo duodenal.

Diatomáceas de tanques do cactário do Jardim Botânico, Guanabara, Brasil

O autor analisou a microflora de três tanques situados no Cactário do Jardim Botânico, Rio de Janeiro, Guanabara. Tanque nº 1. Apresentou desenvolvimento muito intenso da microfibra nos mseses mais frios. As diatomáceas foram muito freqüentes. As espécies indicadoras de saprobidade foram as seguintes: Gomphonema gracile Ehr., pinnularia maior. (kutz) Cleve, Gomphonema parvulum (kutz) Grunow, Navicula mutica Kutz., Pinnularia borealis Ehr., Pinnularia microstauron (Ehr) Cleve, Pinnularia acrospheria Breb., Hantzschia amphioxys (Ehr) Grunow, Nitzschia palea Kutz., Eutonia pectinalis (Kutz) Rabenh. Tanque nº 2. As diatomáceas indicadoras de águas contaminadas, anotamos como segue: Eutonia pectinalis (Kutz) Rabenh., Gomphonema parvulum (kutz) Grunow, Hanstzschia amphioxys (Ehr) Grunow, Navicula mutica Kutz, Pinnularia borealis Ehr., Pinnularia maior (Kutz) Cleve, Pinnularia microstauron (Ehr) Cleve. Tanque nº 3. Foi bastante reduzida a freqüência da microflora. Comparando-o com os tanques nº 1 e 2, as diatomáceas e clorofícias observadas, mostraram-se diminuídas nos meses mais quentes. Encontramos as seguintes espécies oligosaprobias: Eunotia pectinalis (Kutz) Rabenh., Gomphonema gracile ehr., Gomphonema parvulum (Kutz) Grunow Hantzschia amphioxys (Ehr) Grunow, Pinnularia borealis Ehr., Pinnularia maior (Kutz) Cleve. Foram consideradas também as clorofíceas quanto ao regime de saprobidade do material estudado. Eutonia augusta f. crenulata Cleve-Euler e Eutonia veneris var. exsecta Clever-Euler encontradas em nossas amostras, são novas para o Brasil. Um total de 71 espécies foram determinadas de 21 coletas realizadas durante o período de 36 meses.

Utilización de alimentos líquidos para el ganado porcino

La distribución de alimentos líquidos al ganado porcino es un sistema alternativo en el cual el alimento se prepara y distribuye a través de un equipo totalmente informatizado y automatizado. Además de materias primas y piensos permite también el uso de co-productos líquidos de la industria agroalimentaria o productos fermentados. Su utilización evita el coste de su transformación o eliminación permitiendo rebajar el coste de producción del cerdo porque su precio es altamente competitivo. La aplicación de alimentos líquidos fermentados tiene efectos benéficos sobre el epitelio intestinal, la microflora digestiva y el estado sanitario de los lechones, reduciendo la morbilidad y aumentando los resultados de crecimiento. Este sistema de alimentación facilita igualmente la aplicación de programas multifase ajustando diariamente el aporte a las necesidades en nutrientes del cerdo reduciendo la excreción (N, P, metales pesados) y con ello, el riesgo de contaminación medioambiental. The distribution of liquid diets for swine is an alternative system where feed is prepared and distributed by an automatic computerized device. In addition to feedstuffs and complementary dry feed, this system allows the use of fresh or fermented agri-food liquid by-products. These products are cheaper and its use as feed avoids the cost of its transformation or elimination, reducing the environmental load and the cost per kg of pig meat. The fermented liquid feed protects the intestinal epithelium, helps to control digestive microflora and animal health improving piglet growth performance after weaning. This feeding device also helps implementation of multiphase feeding programs, adjusting nutrient supplies to requirements daily, and consequently reducing the nutrient excretion (N, P, heavy metals) and the risk of environmental pollution.

Fungi, bacteria and soil pH: the oxalate-carbonate pathway as a model for metabolic interaction

The oxalatecarbonate pathway involves the oxidation of calcium oxalate to low-magnesium calcite and represents a potential long-term terrestrial sink for atmospheric CO2. In this pathway, bacterial oxalate degradation is associated with a strong local alkalinization and subsequent carbonate precipitation. In order to test whether this process occurs in soil, the role of bacteria, fungi and calcium oxalate amendments was studied using microcosms. In a model system with sterile soil amended with laboratory cultures of oxalotrophic bacteria and fungi, the addition of calcium oxalate induced a distinct pH shift and led to the final precipitation of calcite. However, the simultaneous presence of bacteria and fungi was essential to drive this pH shift. Growth of both oxalotrophic bacteria and fungi was confirmed by qPCR on the frc (oxalotrophic bacteria) and 16S rRNA genes, and the quantification of ergosterol (active fungal biomass) respectively. The experiment was replicated in microcosms with non-sterilized soil. In this case, the bacterial and fungal contribution to oxalate degradation was evaluated by treatments with specific biocides (cycloheximide and bronopol). Results showed that the autochthonous microflora oxidized calcium oxalate and induced a significant soil alkalinization. Moreover, data confirmed the results from the model soil showing that bacteria are essentially responsible for the pH shift, but require the presence of fungi for their oxalotrophic activity. The combined results highlight that the interaction between bacteria and fungi is essential to drive metabolic processes in complex environments such as soil.

Helicobacter pylori in dyspeptic children and adults: endoscopic, bacteriologic and histologic correlations

Using different bacteriological (urease test, Gram staining and culture) and histological (Steiner staining and modified Giemsa staining) techniques, we searched for the presence of Helicobacter pylori in the gastric antrum of 200 dyspeptic Brazilian patients (106 females and 94 males aged 19 days to 81 years). The presence of bacteria was then correlated with the endoscopic and histological findings. H. pylori was present in 59.5 of the population studied. In Brazil, colonization occurs early, involving 37 of the dyspeptic population by 20 years of age. The presence of H. pylori in the gastric antrum was strongly associated with duodenal ulcer (P < 0.001) and a normal endoscopic examination did not exclude the possibility of colonization of the gastric antrum by H. pylori. The most sensitive test was the preformed urease test (89). We conclude that more than one diagnostic method should preferably be used for the detection of H. pylori and that the presence of H. pylori is closely correlated with active chronic gastritis (P < 0.001).

Bacterial overgrowth during treatment with omeprazole compared with cimetidine: a prospective randomised double blind study.

BACKGROUND: Gastric and duodenal bacterial overgrowth frequently occurs in conditions where diminished acid secretion is present. Omeprazole inhibits acid secretion more effectively than cimetidine and might therefore more frequently cause bacterial overgrowth. AIM: This controlled prospective study compared the incidence of gastric and duodenal bacterial overgrowth in patients treated with omeprazole or cimetidine. METHODS: 47 outpatients with peptic disease were randomly assigned to a four week treatment regimen with omeprazole 20 mg or cimetidine 800 mg daily. Gastric and duodenal juice were obtained during upper gastrointestinal endoscopy and plated for anaerobic and aerobic organisms. RESULTS: Bacterial overgrowth (> or = 10(5) cfu/ml) was present in 53% of the patients receiving omeprazole and in 17% receiving cimetidine (p < 0.05). The mean (SEM) number of gastric and duodenal bacterial counts was 6.0 (0.2) and 5.0 (0.2) respectively in the omeprazole group and 4.0 (0.2) and 4.0 (0.1) in the cimetidine group (p < 0.001 and < 0.01; respectively). Faecal type bacteria were found in 30% of the patients with bacterial overgrowth. Basal gastric pH was higher in patients treated with omeprazole compared with cimetidine (4.2 (0.5) versus 2.0 (0.2); p < 0.001) and in patients with bacterial overgrowth compared with those without bacterial overgrowth (5.1 (0.6) versus 2.0 (0.1); p < 0.0001). The nitrate, nitrite, and nitrosamine values in gastric juice did not increase after treatment with either cimetidine or omeprazole. Serum concentrations of vitamin B12, beta carotene, and albumin were similar before and after treatment with both drugs. CONCLUSIONS: These results show that the incidence of gastric and duodenal bacterial overgrowth is considerably higher in patients treated with omeprazole compared with cimetidine. This can be explained by more pronounced inhibition of gastric acid secretion. No patient developed signs of malabsorption or an increase of N-nitroso compounds. The clinical significance of these findings needs to be assessed in studies with long-term treatment with omeprazole, in particular in patients belonging to high risk groups such as HIV infected and intensive care units patients.

Microbial flora variations in the respiratory tract of mice

A stable microbial system in the respiratory tract acts as an important defense mechanism against pathogenic microorganisms. Perturbations in this system may allow pathogens to establish. In an ecological environment such as the respiratory tract, there are many diverse factors that play a role in the establishment of the indigenous flora. In the present work we studied the normal microbial flora of different areas of the respiratory tract of mice and their evolution from the time the mice were born. Our interest was to know which were the dominant groups of microorganisms in each area, which were the first capable of colonizing and which dominated over time to be used as probiotic microorganisms. Our results show that Gram negative facultatively anaerobic bacilli and strict anaerobic microorganisms were the last ones to appear in the bronchia, while aerobic and Gram positive cocci were present in all the areas of the respiratory tract. The number of facultative aerobes and strict anaerobes were similar in the nasal passage, pharynx instilled and trachea, but lower in bronchia. The dominant species were Streptococcus viridans and Staphylococcus saprophyticcus, followed by S. epidermidis, Lactobacilli and S. cohnii I which were present on every studied days but at different proportions. This paper is the first part of a research topic investigating the protective effect of the indigenous flora against pathogens using the mice as an experimental model.

Epidemiology of small intestinal atresia in Europe: a register-based study.

BACKGROUND: The epidemiology of congenital small intestinal atresia (SIA) has not been well studied. This study describes the presence of additional anomalies, pregnancy outcomes, total prevalence and association with maternal age in SIA cases in Europe. METHODS: Cases of SIA delivered during January 1990 to December 2006 notified to 20 EUROCAT registers formed the population-based case series. Prevalence over time was estimated using multilevel Poisson regression, and heterogeneity between registers was evaluated from the random component of the intercept. RESULTS: In total 1133 SIA cases were reported among 5126, 164 registered births. Of 1044 singleton cases, 215 (20.6%) cases were associated with a chromosomal anomaly. Of 829 singleton SIA cases with normal karyotype, 221 (26.7%) were associated with other structural anomalies. Considering cases with normal karyotype, the total prevalence per 10 000 births was 1.6 (95% CI 1.5 to 1.7) for SIA, 0.9 (95% CI 0.8 to 1.0) for duodenal atresia and 0.7 (95% CI 0.7 to 0.8) for jejunoileal atresia (JIA). There was no significant trend in SIA, duodenal atresia or JIA prevalence over time (RR=1.0, 95% credible interval (CrI): 1.0 to 1.0 for each), but SIA and duodenal atresia prevalence varied by geographical location (p=0.03 and p=0.04, respectively). There was weak evidence of an increased risk of SIA in mothers aged less than 20 years compared with mothers aged 20 to 29 years (RR=1.3, 95% CrI: 1.0 to 1.8). CONCLUSION: This study found no evidence of a temporal trend in the prevalence of SIA, duodenal atresia or JIA, although SIA and duodenal atresia prevalence varied significantly between registers.