Cervicovaginal aerobic microflora of women with spontaneous abortion or preterm delivery in Araraquara-Brazil

Rotina bacteriológica do conteúdo vaginal e cervical de 22 mulheres com histórico de aborto recente ou ruptura precoce das membranas foi realizada. Chlamydia trachomatis, Streptococcus pyogenes, Streptococcus agalactiae, Candida sp e Gardnerella vaginalis foram isolados em 54,5% (12) das pacientes. Apesar de Ureaplasma urealyticum ter sido frequentemente encontrado (45,5%), somente em 5 das 22 mulheres foi o único microrganismo presente nos materiais analisados. Esses resultados chamam a atenção para a importância de investigação quantitativa bem como qualitativa da microbiota genital em gestantes, tendo em vista ter consequências na gestação.

Fermentation and epiphytic microflora dynamics in sugar cane silage

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

Use of cecal microflora cultured under aerobic or anaerobic conditions in the control of experimental infection of chicks with Salmonella Enteritidis

One-day-old broiler chicks received cecal microflora (CM) cultured under aerobic, anaerobic conditions, or both (mixed) and were then infected with Salmonella Enteritidis, in order to compare the efficacy of these different types of culture in terms of the number of chicks infected, cecal colonization and faecal excretion of the challenging bacteria. Regardless of culture type, CM always led to a smaller number of S. Enteritidis for any of the parameters studied compared to untreated chicks. Aerobic CM demonstrated better efficacy in reducing the number of infected chicks and cecal colonization by S. Enteritidis, followed by mixed CM. No difference was observed in faecal excretion of S. Enteritidis between the chicks that received different types of CM culture. (C) 2002 Elsevier B.V. B.V. All rights reserved.

Use of Pyrosequencing and Denaturing Gradient Gel Electrophoresis to Examine the Effects of Probiotics and Essential Oil Blends on Digestive Microflora in Broilers Under Mixed Eimeria Infection

A protective digestive microflora helps prevent and reduce broiler infection and colonization by enteropathogens. In the current experiment, broilers fed diets supplemented with probiotics and essential oil (EO) blends were infected with a standard mixed Eimeria spp. to determine effects of performance enhancers on ileal and cecal microbial communities (MCs). Eight treatment groups included four controls (uninfected-unmedicated [UU], unmedicated-infected, the antibiotic BMD plus the ionophore Coban as positive control, and the ionophore as negative control), and four treatments (probiotics BC-30 and Calsporin; and EO, Crina Poultry Plus, and Crina PoultryAF). Day-old broilers were raised to 14 days in floor pens on used litter and then were moved to Petersime batteries and inoculated at 15 days with mixed Eimeria spp. Ileal and cecal samples were collected at 14 days and 7 days postinfection. Digesta DNA was subjected to pyrosequencing for sequencing of individual cecal bacteria and denaturing gradient gel electrophoresis (DGGE) for determination of changes in ileal and cecal MC according to percentage similarity coefficient (%SC). Pyrosequencing is very sensitive detecting shifts in individual bacterial sequences, whereas DGGE is able to detect gross shifts in entire MC. These combined techniques offer versatility toward identifying feed additive and mild Eimeria infection modulation of broiler MC. Pyrosequencing detected 147 bacterial species sequences. Additionally, pyrosequencing revealed the presence of relatively low levels of the potential human enteropathogens Campylobacter sp. and four Shigella spp. as well as the potential poultry pathogen Clostridiun perfringens. Pre- and postinfection changes in ileal (56%SC) and cecal (78.5%SC) DGGE profiles resulted from the coccidia infection and with increased broiler age. Probiotics and EO changed MC from those seen in UU ilea and ceca. Results potentially reflect the performance enhancement above expectations in comparison to broilers not given the probiotics or the specific EO blends as feed supplements.

Effect of the consumption of a symbiotic shake on the intestinal microflora of erdely people

The consumption of foods containing probiotic and prebiotic ingredients is growing consistently every year, and in view of the limited number of studies investigating their effect in the elderly. Objective The objective of this study was to evaluate the effect of the consumption of a symbiotic shake containing Lactobacillus acidophilus, Bifidobacterium bifidum and fructooligosaccharides on glycemia and cholesterol levels in elderly people. Methods A randomized, double-blind, placebo-controlled study was conducted on twenty volunteers (ten for placebo group and ten for symbiotic group), aged 50 to 60 years. The criteria for inclusion in the study were: total cholesterol > 200 mg/dL; triglycerides > 200 mg/dL and glycemia > 110 mg/dL. Over a total test period of 30 days, 10 individuals (the symbiotic group) consumed a daily dose of 200 mL of a symbiotic shake containing 108 UFC/mL Lactobacillus acidophilus, 108 UFC/mL Bifidobacterium bifidum and 2 g oligofructose, while 10 other volunteers (the placebo group) drank daily the same amount of a shake that did not contain any symbiotic bacteria. Blood samples were collected 15 days prior to the start of the experiment and at 10-day intervals after the beginning of the shake intake. The standard lipid profile (total cholesterol, triglycerides and HDL cholesterol) and glycemia, or blood sugar levels, were evaluated by an enzyme colorimetric assay. Results The results of the symbiotic group showed a non-significant reduction (P > 0.05) in total cholesterol and triglycerides, a significant increase (P < 0.05) in HDL cholesterol and a significant reduction (P < 0.05) in fasting glycemia. No significant changes were observed in the placebo group. Conclusion The consumption of symbiotic shake resulted in a significant increase in HDL and a significant decrease of glycemia.

Efeito da penicilina G a cada três semanas sobre o surgimento de Streptococcus viridans resistentes à penicilina na microflora oral

FUNDAMENTO: Penicilina G benzatina a cada 3 semanas é o protocolo padrão para a profilaxia secundária para febre reumática recorrente. OBJETIVO: Avaliar o efeito da penicilina G benzatina em Streptococcus sanguinis e Streptococcus oralis em pacientes com doença valvular cardíaca, devido à febre reumática com recebimento de profilaxia secundária. MÉTODOS: Estreptococos orais foram avaliados antes (momento basal) e após 7 dias (7º dia) iniciando-se com penicilina G benzatina em 100 pacientes que receberam profilaxia secundária da febre reumática. Amostras de saliva foram avaliadas para verificar a contagem de colônias e presença de S. sanguinis e S. oralis. Amostras de saliva estimulada pela mastigação foram serialmente diluídas e semeadas em placas sobre agar-sangue de ovelhas seletivo e não seletivo a 5% contendo penicilina G. A identificação da espécie foi realizada com testes bioquímicos convencionais. Concentrações inibitórias mínimas foram determinadas com o Etest. RESULTADOS: Não foram encontradas diferenças estatísticas da presença de S. sanguinis comparando-se o momento basal e o 7º dia (p = 0,62). No entanto, o número existente de culturas positivas de S. oralis no 7º dia após a Penicilina G benzatina apresentou um aumento significativo em relação ao valor basal (p = 0,04). Não houve diferença estatística existente entre o momento basal e o 7º dia sobre o número de S. sanguinis ou S. oralis UFC/mL e concentrações inibitórias medianas. CONCLUSÃO: O presente estudo mostrou que a Penicilina G benzatina a cada 3 semanas não alterou a colonização por S. sanguinis, mas aumentou a colonização de S. oralis no 7º dia de administração. Portanto, a susceptibilidade do Streptococcus sanguinis e Streptococcus oralis à penicilina G não foi modificada durante a rotina de profilaxia secundária da febre reumática utilizando a penicilina G.

Effetto della temperatura di conservazione del latte, dalla raccolta all'affioramento, sulla microflora del latte, del sieroinnesto e del formaggio Trentingrana

This PhD research is part of a project addressed to improve the quality of Grana Trentino production. The objectives were to evaluated if milk storage and collection procedures may affect cheese-making technology and quality. Actually the milk is collected and delivered to the cheese factory just after milking in 50 L cans without refrigeration or in tanks cooled at 18 °C. This procedure is expensive (two deliveries each day) and the milk quality is difficult to preserve as temperatures are not controlled. The milk refrigeration at the farm could allow a single delivery to the dairy. Therefore it could be a good strategy to preserve raw milk quality and reduce cheese spoilage. This operation may, however, have the drawbacks of favouring the growth of psychrotrophic bacteria and changing the aptitude of milk to coagulation. With the aim of studying the effect on milk and cheese of traditional and new refrigerated technologies of milk storage, two different collection and creaming technologies were compared. The trials were replicated in three cheese factories manufacturing Grana Trentino. Every cheese-making day, about 1000 milk liters were collected from always the same two farms in the different collection procedures (single or double). Milk was processed to produce 2 wheels of Grana trentino every day. During the refrigerated trials, milk was collected and stored at the farm in a mixed tank at 12 or 8 °C and then was carried to the dairy in truck once a day. 112 cheese making day were followed: 56 for traditional technology and 56 for the refrigerated one. Each one of these two thechnologies lead to different ways of creaming: long time in the traditional one and shorter in the new one. For every cheese making day we recorded time, temperatures and pH during the milk processing to cheese. Whole milk before ceraming, cream and skim milk after creaming, vat milk and whey were sampled during every cheese-making day for analysis. After 18 months ripening we opened 46 cheese wheels for further chemical and microbiological analyses. The trials were performed with the aim of: 1 estimate the effect of storage temperatures on microbial communities, physico-chemical or/and rheological differences of milk and skim milk after creaming. 2 detect by culture dependent (plate counts) and indipendent (DGGE) methodolgies the microbial species present in whole, skimmed milk, cream and cheese sampled under the rind and in the core; 3 estimate the physico-chemical characteristics, the proteolytic activity, the content of free aminoacids and volatile compounds in 18 months ripened Grana Trentino cheeses from different storing and creaming of milk technologies. The results presented are remarkable since this is the first in-deep study presenting microbiological and chemical analysis of Grana Trentino that even if belonging to Grana Padano Consortium, it is clearly different in the milk and in the manufacturing technology.

Caratterizzazione della microflora spontanea di un alimento fermentato vegano

Tra i prodotti vegani più richiesti vi sono i “formaggi” vegani, alimenti fermentati a base di frutta secca o ottenuti attraverso trattamenti su latte di mandorla e latte di soia, e successivamente fermentati. Nella mia attività ho caratterizzato un alimento fermentato vegano e studiato la successione microbica durante la fermentazione di un “formaggio” ottenuto partendo da anacardi e preparato in maniera artigianale. Oltre agli aspetti microbiologici, ho analizzato anche alcune caratteristiche fisico chimiche. Durante il processo di produzione gli anacardi vengono messi in ammollo per 8 ore a temperatura ambiente e, successivamente, i semi vengono scolati e risciacquati sotto acqua corrente. Gli anacardi vengono poi addizionati di acqua e microrganismi probiotici e tritati in un mixer fino al raggiungimento di una crema omogenea. A questo punto il prodotto viene lasciato riposare a temperatura ambiente per 48 ore durante le quali ha luogo la fermentazione e poi addizionato di ingredienti. Le indagini chimico fisiche effettuate hanno evidenziato che il pH si mostra già basso prima dell’inizio della fermentazione vera e propria e scende a 4.5 dopo 48 ore di riposo a causa dell’accumulo di acidi organici, ed in particolare di acido lattico e acetico che indicando un’attività fermentativa condotta dai batteri lattici. Le analisi microbiologiche hanno confermato che l’effettivo agente di fermentazione era costituito da questi batteri che sono stati identificati a livello molecolare. Le specie identificate due eterofermentanti (Weissella e Leuconostoc), presenti soprattutto nelle prime fasi della fermentazione, ed una omofermentante (Pediococcus), prende il sopravvento mano a mano che la fermentazione procede. Il lavoro svolto ha permesso di ottenere alcune importanti informazioni per la produzione industriale di un “formaggio” vegano fermentato. Il processo studiato presenta numerosi punti di rischio che devono essere presi in considerazione prima di poter giungere alla messa a punto di un prodotto definitivo.

The vaginal microflora in relation to gingivitis

BACKGROUND: Gingivitis has been linked to adverse pregnancy outcome (APO). Bacterial vaginosis (BV) has been associated with APO. We assessed if bacterial counts in BV is associated with gingivitis suggesting a systemic infectious susceptibilty. METHODS: Vaginal samples were collected from 180 women (mean age 29.4 years, SD +/- 6.8, range: 18 to 46), and at least six months after delivery, and assessed by semi-quantitative DNA-DNA checkerboard hybridization assay (74 bacterial species). BV was defined by Gram stain (Nugent criteria). Gingivitis was defined as bleeding on probing at >or= 20% of tooth sites. RESULTS: A Nugent score of 0-3 (normal vaginal microflora) was found in 83 women (46.1%), and a score of > 7 (BV) in 49 women (27.2%). Gingivitis was diagnosed in 114 women (63.3%). Women with a diagnosis of BV were more likely to have gingivitis (p = 0.01). Independent of gingival conditions, vaginal bacterial counts were higher (p < 0.001) for 38/74 species in BV+ in comparison to BV- women. Counts of four lactobacilli species were higher in BV- women (p < 0.001). Independent of BV diagnosis, women with gingivitis had higher counts of Prevotella bivia (p < 0.001), and Prevotella disiens (p < 0.001). P. bivia, P. disiens, M. curtisii and M. mulieris (all at the p < 0.01 level) were found at higher levels in the BV+/G+ group than in the BV+/G- group. The sum of bacterial load (74 species) was higher in the BV+/G+ group than in the BV+/G- group (p < 0.05). The highest odds ratio for the presence of bacteria in vaginal samples (> 1.0 x 104 cells) and a diagnosis of gingivitis was 3.9 for P. bivia (95% CI 1.5-5.7, p < 0.001) and 3.6 for P. disiens (95%CI: 1.8-7.5, p < 0.001), and a diagnosis of BV for P. bivia (odds ratio: 5.3, 95%CI: 2.6 to 10.4, p < 0.001) and P. disiens (odds ratio: 4.4, 95% CI: 2.2 to 8.8, p < 0.001). CONCLUSION: Higher vaginal bacterial counts can be found in women with BV and gingivitis in comparison to women with BV but not gingivitis. P. bivia and P. disiens may be of specific significance in a relationship between vaginal and gingival infections.

Subgingival microflora in inflammatory bowel disease patients with untreated periodontitis

OBJECTIVE To analyze the subgingival microflora composition of inflammatory bowel disease (IBD) patients with untreated chronic periodontitis and compare them with systemically healthy controls also having untreated chronic periodontitis. METHOD Thirty IBD patients [15 with Crohn's disease (CD) and 15 with ulcerative colitis (UC)] and 15 control individuals participated in the study. All patients had been diagnosed with untreated chronic periodontitis. From every patient, subgingival plaque was collected from four gingivitis and four periodontitis sites with paper points. Samples from the same category (gingivitis or periodontitis) in each patient were pooled together and stored at -70 °C until analysis using a checkerboard DNA-DNA hybridization technique for 74 bacterial species. RESULTS Multiple-comparison analysis showed that the groups differed in bacterial counts for Bacteroides ureolyticus, Campylobacter gracilis, Parvimonas micra, Prevotella melaninogenica, Peptostreptococcus anaerobius, Staphylococcus aureus, Streptococcus anginosus, Streptococcus intermedius, Streptococcus mitis, Streptococcus mutans, and Treponema denticola (P<0.001). CD patients had significantly higher levels of these bacteria than UC patients either in gingivitis or in periodontitis sites (P<0.05). CD patients harbored higher levels of P. melaninogenica, S. aureus, S. anginosus, and S. mutans compared with controls both at gingivitis and at periodontitis sites (P<0.05). UC patients harbored higher levels of S. aureus (P=0.01) and P. anaerobius (P=0.05) than controls only in gingivitis sites. CONCLUSION Our study showed that even with similar clinical periodontal parameters, IBD patients harbor higher levels of bacteria that are related to opportunistic infections in inflamed subgingival sites that might be harmful for the crucial microbe-host interaction.