Campylobacter sp como perigo biológico na produção de peito de frango maturado

Pós-graduação em Microbiologia Agropecuária - FCAV

Avaliação comparativa in vivo da eficácia do óleo de melaleuca, clorexidina e listerine sobre streptococcus mutans e microrganismos totais na saliva

Objective: To compare the efficacy of the mouthwashes 0.12% chlorhexidine, Listerine, and 0.5% and 2% Melaleuca Alternifolia oil against the salivary levels of Streptococcus mutans and total microorganisms. Methods: This study was double-blind controlled and paired clinical assay. Twenty-six volunteers aged 21 to 35 years old were enrolled. At baseline, 1 mL of unstimulated saliva was collected from each subject, 1 and 15 min after mouthrinsing with the following solutions: sterile distilled water, 0.12% chlorhexidine digluconate, Listerine (©Johnson & Johnson do Brasil), 0.5% and 2% concentrations of Melaleuca Alternifolia (Sigma-Aldrich). The volunteers used all the evaluated mouthrinses with a 15-day interval between the solutions. Immediately after rinsing, saliva was collected and serial dilutions were performed, followed by plating in blood agar culture medium for growth of total microorganisms and SB-20 (Sucrose-Bacitracin agar) for growth of S. mutans, and incubation at 37 °C for 48 h in microaerophilia. After incubation, the number of colonies was counted and expressed as colony forming units (UFC/mL). Results: Chlorhexidine showed antimicrobial action by reducing total microorganisms and S. mutans, while the action of 0.5% Melaleuca Alternifolia was similar to that of distilled water. Listerine and 2% Melaleuca Alternifolia oil reduced total microbial counts by 11% and 9% respectively, and S. mutans by 20% and 11%. Conclusion: A single rinse with 0.12% chlorhexidine is effective in reducing the levels of total microorganisms and S. mutans present in saliva. Under the same testing conditions, Listerine and 0.5% and 2% Melaleuca Alternifolia oil presented lower efficacy than chlorhexidine.

Virginiamycin via mineral supplementation and sward height in growing Nelore young bulls

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

Application of an active attachment model as a high-throughput demineralization biofilm model

Objectives: To investigate the potential of an active attachment biofilm model as a highthroughput demineralization biofilm model for the evaluation of caries-preventive agents. Methods: Streptococcus mutans UA159 biofilms were grown on bovine dentine discs in a highthroughput active attachment model. Biofilms were first formed in a medium with high buffer capacity for 24 h and then subjected to various photodynamic therapies (PACT) using the combination of Light Emitting Diodes (LEDs, Biotable (R)) and Photogem (R). Viability of the biofilms was evaluated by plate counts. To investigate treatment effects on dentine lesion formation, the treated biofilms were grown in a medium with low buffer capacity for an additional 24 h. Integrated mineral loss (IML) and lesion depth (LD) were assessed by transversal microradiography. Calcium release in the biofilm medium was measured by atomic absorption spectroscopy. Results: Compared to the water treated control group, significant reduction in viability of S. mutans biofilms was observed when the combination of LEDs and Photogem (R) was applied. LEDs or Photogem (R) only did not result in biofilm viability changes. Similar outcomes were also found for dentine lesion formation. Significant lower IML and LD values were only found in the group subjected to the combined treatment of LEDs and Photogem (R). There was a good correlation between the calcium release data and the IML or LD values. Conclusions: The high-throughput active attachment biofilm model is applicable for evaluating novel caries-preventive agents on both biofilm and demineralization inhibition. PACT had a killing effect on 24 h S. mutans biofilms and could inhibit the demineralization process. (C) 2011 Elsevier Ltd. All rights reserved.

Validation of a new proposal of quantitative sperm culture for industrialized bull semen

The Brazilian Ministry of Agriculture (MAPA) has discussed the mandatory culture of industrialized semen, both to ensure biosefety, and to prevent in vitro fertilization problems caused by oocyte contamination with ubiquitous and opportunistic bacteria from preputial microbiota. Pour plate, a quantitative technique recommended by the World Organization for Animal Health (OIE), is operationally difficult and costly for routine analysis in Artificial Insemination Centers (AICs). The objectives of this study were to evaluate and validate viable superficial bacteria counts (VSBC), in CFU/mL, compared with pour plate counts, in industrialized bull semen samples from AICs. Semen straws from Projeto Hungria - MAPA bulls were used. VSBC and pour plate were carried out in parallel in serial dilutions of the samples, from 10(-1) to 10(-5). CFU/mL means or medians recorded in each dilution and technique were compared, and no statistical differences were observed between the two techniques regarding the quantification of bacteria in CFU/mL, suggesting that it may be possible to replace pour plate for CBSV, a cheaper and more practical technique.

Comparison of Ozone and Chlorine in Low Concentrations as Sanitizing Agents of Chicken Carcasses in the Water Immersion Chiller

The aim of this study was to investigate the effects of the use of chlorine or ozone as sanitizing agents in the water of chicken immersion chilling, using the residual levels usually applied in Brazil (1.5 ppm), comparing the effects of these treatments on the microbiological, physicochemical, and sensory characteristics of carcasses. Chicken carcasses were chilled in water (4 degrees C) with similar residual levels of ozone and chlorine until reaching temperatures below 7 degrees C (around 45 min). The stability of carcasses was assessed during 15 days of storage at 2 +/- 1 degrees C. Microbiological, surface color (L*, a*, b* parameters), pH value, lipid oxidation (thiobarbituric acid reactive substances index), and sensory evaluation (on a 9-point hedonic scale for odor and appearance) analyses were carried out. The presence of Salmonella was not detected, coagulase-positive staphylococci counts were below 10(2) CFU/ml of rinse fluid, and Escherichia coil and total coliform counts were below 10(5) CFU/ml of rinse fluid until the end of the storage period for both treatments. Psychrotrophic microorganism counts did not differ (P > 0.05) between chlorine and ozone treatments, and both values were near 10(9) CFU/ml of rinse fluid after 15 days at 4 +/- 1 degrees C. pH values did not differ between treatments (P > 0.05) or during the storage period (P > 0.05). In addition, neither chlorine nor ozone treatment showed differences (P > 0.05) in the lipid oxidation of carcasses; however, the thiobarbituric acid reactive substances index of both treatments increased (P <= 0.05) during the storage period, reaching values of approximately 0.68 mg of malonaldehyde per kg. Samples from both treatments did not differ (P > 0.05) in their acceptance scores for odor and overall appearance, but in the evaluation of color, ozone showed an acceptance score significantly higher (P <= 0.05) than that for the chlorine treatment. In general, under the conditions tested, ozone showed results similar to the results for chlorine in the disinfection of chicken carcasses in the immersion chilling, which may indicate its use as a substitute for chlorine in poultry slaughterhouses.

Lactobacillus acidophilus CRL 1014 improved “gut health” in the SHIME® reactor

Abstract Background How to maintain “gut health” is a goal for scientists throughout the world. Therefore, microbiota management models for testing probiotics, prebiotics, and synbiotics have been developed. Methods The SHIME® model was used to study the effect of Lactobacillus acidophilus 1014 on the fermentation pattern of the colon microbiota. Initially, an inoculum prepared from human feces was introduced into the reactor vessels and stabilized over 2-wk using a culture medium. This stabilization period was followed by a 2-wk control period during which the microbiota was monitored. The microbiota was then subjected to a 4-wk treatment period by adding 5 mL of sterile peptone water with L. acidophilus CRL1014 at the concentration of 108 CFU/mL to vessel one (the stomach compartment). Plate counts, Denaturing Gradient Gel Electrophoresis (DGGE), short-chain fatty acid (SCFA) and ammonium analyses were carried out for monitoring of the microbial community from the colon compartments. Results A significant increase (p < 0.01) in the Lactobacillus spp. and Bifidobacterium spp. populations was observed during the treatment period. The DGGE obtained showed changes in the lactobacilli community from the colon compartments of the SHIME® reactor. The (SCFA) concentration increased (p < 0.01) during the treatment period, due mainly to significant increased levels of acetic, butyric, and propionic acids. However, ammonium concentrations decreased during the same period (p < 0.01). Conclusions This study showed the beneficial influence of L. acidophilus CRL 1014 on microbial metabolism and lactobacilli community composition for improving human health.

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.

Microbial colonization patterns predict the outcomes of surgical treatment of intrabony defects

AIM: To explore the impact of bacterial load and microbial colonization patterns on the clinical outcomes of periodontal surgery at deep intrabony defects. MATERIALS AND METHODS: One hundred and twenty-two patients with advanced chronic periodontitis and at least one intrabony defect of >3 mm were recruited in 10 centres. Before recruitment, the infection control phase of periodontal therapy was completed. After surgical access and debridement, the regenerative material was applied in the test subjects, and omitted in the controls. At baseline and 1 year following the interventions, clinical attachment levels (CAL), pocket probing depths (PPD), recession (REC), full-mouth plaque scores and full-mouth bleeding scores were assessed. Microbial colonization of the defect-associated pocket was assessed using a DNA-DNA checkerboard analysis. RESULTS: Total bacterial load and counts of red complex bacteria were negatively associated with CAL gains 1 year following treatment. The probability of achieving above median CAL gains (>3 mm) was significantly decreased by higher total bacterial counts, higher red complex and T. forsythensis counts immediately before surgery. CONCLUSIONS: Presence of high bacterial load and specific periodontal pathogen complexes in deep periodontal pockets associated with intrabony defects had a significant negative impact on the 1 year outcome of surgical/regenerative treatment.

Maternal tobacco smoking and decreased leukocytes, including dendritic cells, in neonates

Maternal smoking in pregnancy is associated with respiratory diseases in the offspring, possibly due to prenatal influences on the developing immune system. We investigated whether maternal smoking in pregnancy was associated with cord blood leukocyte numbers, including precursor dendritic cells, adjusting for concomitant factors. In a prospective healthy birth cohort study, total leukocyte counts were reduced in neonates of smoking mothers [10.7 (8.4-13.0), n=14] compared with nonexposed infants [14.7 (13.7-15.7), n=74, p=0.002] [geometric mean cells x 10(3)/microL (95% confidence interval)]. All leukocyte subsets were decreased, most prominently segmented neutrophils [4.3 (2.8-5.7) versus 6.2 (5.5-6.8), p=0.021], lymphocytes [3.8 (2.9-4.8) versus 5.0 (4.5-5.6), p=0.036], and myeloid precursor dendritic cells [12.7 cells/microL (9.1-17.8) versus 18.3 (15.8-21.2), p=0.055]. These differences persisted after adjustment for possible confounders. Predictors of myeloid precursor dendritic cell numbers in multivariable models were maternal smoking (-5.1 cells/microL, p=0.042), age (-0.5 cells/microL/y, p=0.035), and, marginally, asthma (+8.1 cells/microL, p=0.075). The decrease of all leukocytes in neonates of smoking mothers could be clinically significant and suggests a decreased cell production, increased peripheral recruitment, or retention in bone marrow. Given the importance of dendritic cells in early immune responses, their decrease might reflect an impact of maternal smoking on the developing fetal immune system.

Development of a highly sensitive and specific assay to detect Staphylococcus aureus in bovine mastitic milk

Diagnosis of udder infections with Staphylococcus aureus by bacteriological milk testing of quarter milk samples is often not satisfactory. To get reliable results, repeated sampling is necessary, which is normally too expensive. Therefore, we developed a test that allows the highly specific detection of Staph. aureus in bovine milk samples at very low concentrations. It is based on a fast procedure to prepare bacteria from milk, followed by DNA extraction and quantitative PCR. The whole analysis is done within 5 h. For clinical milk samples, the analytical sensitivity of the assay was 50.7 times and 507 times higher than conventional bacteriology with 100 and 10 microL, respectively. The diagnostic specificity was 100%. The test is further characterized by a low intra- and interassay variability as well as by a good recovery of Staph. aureus from raw milk. Furthermore, a high correlation (R = 0.925) between the agar plate counts and the quantitative PCR methodology over the whole range of measurement was found. In addition, our test revealed considerably more positive results than bacteriology. Due to its favorable properties, the assay might become an important diagnostic tool in the context of bovine mastitis caused by Staph. aureus.

The impact of a low-frequency chlorhexidine rinsing schedule on the subgingival microbiota (the TEETH clinical trial)

BACKGROUND: Information on the efficacy of chlorhexidine (CHX) rinsing on the subgingival microbiota is limited. This study tested if intermittent CHX rinsing over 5 years had an impact on the subgingival microbiota. METHODS: Subgingival plaque samples were analyzed by the checkerboard DNA-DNA hybridization method in a double-blind randomized CHX rinse study. RESULTS: A total of 210 subjects were included. The mean age of the subjects was 71.7 (+/- 4.1) years, and 56.2% were women. Evidence of alveolar bone loss was found in 39% of subjects. Bacterial loads were not correlated significantly with probing depth. At year 5, subjects in the CHX rinse group with no evidence of bone loss presented with lower total bacterial counts than control subjects with no bone loss. The levels of the following bacteria were significantly lower in the CHX group: Lactobacillus acidophilus (P <0.05), Eikenella corrodens (P <0.05), Fusobacterium nucleatum sp. nucleatum (P <0.01), Treponema denticola (P <0.05), Leptotrichia buccalis (P <0.05), and Eubacterium saburreum (P <0.05). No differences in bacterial loads were found between CHX and control rinse subjects with alveolar bone loss. CONCLUSIONS: Older subjects with or without periodontitis carry a large variety of bacteria associated with periodontitis. Intermittent rinsing with CHX may provide a preventive benefit in reducing levels of bacteria but only in subjects without alveolar bone loss.

Does pregnancy have an impact on the subgingival microbiota?

BACKGROUND: We investigated clinical and subgingival microbiologic changes during pregnancy in 20 consecutive pregnant women > or =18 years not receiving dental care. METHODS: Bacterial samples from weeks 12, 28, and 36 of pregnancy and at 4 to 6 weeks postpartum were processed for 37 species by checkerboard DNA-DNA hybridization. Clinical periodontal data were collected at week 12 and at 4 to 6 weeks postpartum, and bleeding on probing (BOP) was recorded at sites sampled at the four time points. RESULTS: The mean BOP at week 12 and postpartum was 40.1% +/- 18.2% and 27.4% +/- 12.5%, respectively. The corresponding mean BOP at microbiologic test sites was 15% (week 12) and 21% (postpartum; not statistically significant). Total bacterial counts decreased between week 12 and postpartum (P <0.01). Increased bacterial counts over time were found for Neisseria mucosa (P <0.001). Lower counts (P <0.001) were found for Capnocytophaga ochracea, Capnocytophaga sputigena, Eubacterium saburreum, Fusobacterium nucleatum naviforme, Fusobacterium nucleatum polymorphum, Leptotrichia buccalis, Parvimonas micra (previously Peptostreptococcus micros or Micromonas micros), Prevotella intermedia, Prevotella melaninogenica, Staphylococcus aureus, Streptococcus anginosus, Streptococcus intermedius, Streptococcus mutans, Streptococcus oralis, Streptococcus sanguinis, Selenomonas noxia, and Veillonella parvula. No changes occurred between weeks 12 and 28 of pregnancy. Counts of Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), Porphyromonas gingivalis, Tannerella forsythia (previously T. forsythensis), and Treponema denticola did not change. Counts of P. gingivalis and T. forsythia at week 12 were associated with gingivitis (P <0.001). CONCLUSIONS: Subgingival levels of bacteria associated with periodontitis did not change. P. gingivalis and T. forsythia counts were associated with BOP at week 12. A decrease was found in 17 of 37 species from week 12 to postpartum. Only counts of N. mucosa increased.

Mechanical non-surgical treatment of peri-implantitis: a double-blind randomized longitudinal clinical study. I: clinical results

BACKGROUND: Peri-implantitis is a frequent finding in patients with dental implants. The present study compared two non-surgical mechanical debridement methods of peri-implantitis. MATERIAL AND METHODS: Thirty-seven subjects (mean age 61.5; S.D+/-12.4), with one implant each, demonstrating peri-implantitis were randomized, and those treated either with titanium hand-instruments or with an ultrasonic device were enrolled. Data were obtained before treatment, and at 1, 3, and 6 months. Parametric and non-parametric statistics were used. RESULTS: Thirty-one subjects completed the study. The mean bone loss at implants in both groups was 1.5 mm (SD +/-1.2 mm). No group differences for plaque or gingival indices were found at any time point. Baseline and 6-month mean probing pocket depths (PPD) at implants were 5.1 and 4.9 mm (p=0.30) in both groups. Plaque scores at treated implants decreased from 73% to 53% (p<0.01). Bleeding scores also decreased (p<0.01), with no group differences. No differences in the total bacterial counts were found over time. Higher total bacterial counts were found immediately after treatment (p<0.01) and at 1 week for ultrasonic-treated implants (p<0.05). CONCLUSIONS: No group differences were found in the treatment outcomes. While plaque and bleeding scores improved, no effects on PPD were identified.

Comparison of the immediate effects of gaseous ozone and chlorhexidine gel on bacteria in cavitated carious lesions in children in vivo

Clinical application of ozone gas has been shown to arrest the progression of dentinal caries in children. In this study, we compare the immediate effects of gaseous ozone and chlorhexidine gel on bacteria in cavitated carious lesions in children. Forty children, each with at least two open occlusal carious lesions, were enrolled in the study. Two teeth were chosen randomly. In one lesion, overlying soft biological material was removed, whilst the other lesion was not excavated. Cavities were rinsed with sterile water and dried with air. A standardised sample was taken from the mesial part of each lesion. Then, gaseous ozone (HealOzone) or 1% chlorhexidine gel (Corsodyl) was applied for 30 s on both lesions of 20 children each, and a second sample was taken from the distal part of each lesion. The anaerobic microbiota was cultivated; the number of colony forming units was calculated per milligram sample. The two-sided paired t test showed no significant (P > 0.05) differences in the reduction of total bacterial counts per milligram comparing samples before and after ozone or chlorhexidine application. The tests also showed no statistically significant difference whether the superficial decayed dentine had been removed before ozone or with chlorhexidine treatment or not. It can be concluded that gaseous ozone or chlorhexidine gel application for 30 s to deep occlusal carious cavities had no significant immediate antimicrobial effects whether the superficial decayed layers dentine were removed or not.