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1.
A cross sectional study was conducted to enumerate total viable bacteria (TBC), coliforms, Escherichia coli and Staphylococcus aureus in raw (n = 120) and processed (n = 20) milk from individual farms from three smallholder dairy schemes of Zimbabwe between October, 2009 and February, 2010. Data on management factors were collected using a structured questionnaire. A standard pour plate technique was used to enumerate total viable bacteria, while for coliforms, E. coli and S. aureus, counts were assessed by the spread plate technique. The association of total viable bacterial counts and management factors was assessed using univariable and a linear regression model. The log10 TBC for raw milk differed significantly (P < 0.05) amongst the schemes with the lowest (5.6 ± 4.7 log10 cfu/ml) and highest (6.7 ± 5.8 log10 cfu/ml) recorded from Marirangwe and Nharira respectively. The mean log10 of TBC of processed milk (6.6 ± 6.0 log10 cfu/ml) were marginally higher than those of raw milk (6.4 ± 5.6 log10 cfu/ml) but not significant (P > 0.05). The coliform, E. coli and S. aureus counts for raw milk significantly differed (P < 0.05) amongst the study areas. The variation in TBC, coliforms, E. coli and S. aureus counts amongst the schemes could be attributed to differences in milking hygiene where farms with more access to training and monitoring of microbiological quality of milk had lower counts. Linear regression analysis revealed dairy scheme, delivery time and season of milking as independently associated with increased TBC of raw milk. The high TBC of raw and processed milk generally indicated low levels of milking hygienic practices, and high level of post-processing contamination, respectively. The high TBC, coliform, E. coli and S. aureus counts of both raw and processed milk may present a public health hazard. Thus, educating the farmers on general hygienic practices, quickening the delivery of milk to collection centres, or availing cooling facilities on-farm will improve the microbiological quality and safety of milk.  相似文献   

2.
Previously there was no available information on the levels of indicator bacteria and the prevalence of pathogens in fresh lettuce grown in organic and conventional farms in Spain. A total of 72 lettuce samples (18 farms for 4 repetitions each) for each type of the agriculture were examined in order to assess the bacteriological quality of the lettuces, in particular the prevalence of selected pathogens. The lettuce samples were analyzed for the presence of aerobic mesophilic, psychrotrophic microorganisms, yeasts and moulds, Enterobacteriaceae, mesophilic lactic acid bacteria, Pseudomonas spp. and presumptive Escherichia coli, Salmonella spp. and Listeria monocytogenes. The mean aerobic mesophilic counts (AM) were 6.35 ± 0.69 log10 cfu g−1 and 5.67 ± 0.80 log10 cfu g−1 from organic and conventional lettuce, respectively. The mean counts of psychrotrophic microorganisms were 5.82 ± 1.01 log10 cfu g−1 and 5.41 ± 0.92 log10 cfu g−1 from organic and conventional lettuce, respectively. Yeasts and moulds (YM) mean counts were 4.74 ± 0.83 log10 cfu g−1 and 4.21 ± 0.96 log10 cfu g−1 from organic and conventional lettuce, respectively. Lactic acid bacteria (LAB) were present in low numbers and the mean counts were 2.41 ± 1.10 log10 cfu g−1 and 1.99 ± 0.91 log10 cfu g−1 from organic and conventional lettuce, respectively. Pseudomonas spp. mean counts were 5.49 ± 1.37 log10 cfu g−1 and 4.98 ± 1.26 log10 cfu g−1 in organic and conventional lettuce, respectively. The mean counts for Enterobacteriaceae were 5.16 ± 1.01 log10 cfu g−1 and 3.80 ± 1.53 log10 cfu g−1 in organic and conventional lettuce, respectively. E. coli was detected in 22.2% (16 samples) of organic lettuce and in 12.5% (9 samples) of conventional lettuce. None of the lettuce samples was positive for E. coli O157:H7, L. monocytogenes and Salmonella spp. From the samples analyzed by principal component analysis (PCA) a pattern with two different groups (conventional and organic) can be observed, being the highest difference between both kinds of samples the Enterobacteriaceae count.  相似文献   

3.
This study describes a procedure that allows specific detection and enumeration of viable bacteria in four species of lactic acid bacteria (Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus casei subsp. casei and Lactobacillus acidophilus) and of Bifidobacterium lactis, mixed in fermented milk products. The procedure is based on the combined use of propidium monoazide (PMA), able to distinguish between viable and irreversibly damaged cells, with species-specific quantitative real-time PCR (RTi-PCR). Loss of viability of the species in a fermented milk through storage at 4 °C was similarly (P < 0.05) detected by PMA–RTi-PCR and selective plate counts. Furthermore, comparison of results obtained by both methods showed a Pearson linear correlation of 0.995. The enumeration of viable bacteria by PMA–RTi-PCR could be performed in 3 h, whereas enumeration by selective plate counts required three days. The procedure developed is a fast method for the identification, enumeration and discrimination of viability of lactic acid bacteria and bifidobacteria mixed in fermented milk products.  相似文献   

4.
H. Ölmez  S.D. Temur 《LWT》2010,43(6):964-970
The effects of ozone (2 mg/L), chlorine (100 mg/L) and organic acid (0.25 g/100 g citric acid plus 0.50 g/100 g ascorbic acid) treatments at 10 °C for 2 min on the removal of Escherichia coli and Listeria monocytogenes cells embedded inside biofilms on the surface of lettuce leaves were studied. None of the sanitizing treatments were found effective in removing the bacterial biofilms. Initiation of biofilms was observed after 24 h of incubation. Bacterial cells appeared as individual cells, rather than clusters after 6 h incubation, thus 99.9% reductions in both E. coli and L. monocytogenes counts were achieved with all the three treatments. However, after 48 h incubation, none of the treatments resulted in higher than 90% reduction in microbial counts. Biofilm formation was demonstrated for the 48 h incubated samples with SEM images.  相似文献   

5.
In food industry, biofilms are a source of recalcitrant contaminations, being possible sources of public health problems. The aims of this study were evaluate the disinfectant efficacy of peracetic acid against Pseudomonas aeruginosa biofilms grown on polystyrene surfaces, determining the best method to measuring its disinfecting efficacy. PERAsafe® has been used as peracetic acid donor into medium. Direct colony-forming units (CFUs) counts together with two assays based in optical density measurement have been tested. Starting from bacterial suspensions (∼105), the log10 values obtained were 4.34 ± 0.20, 2.60 ± 0.77 and 0.00 for 0.4%, 0.8% and 1.61%, PERAsafe® concentrations, respectively, showing that this compound achieved the 100% killing efficacy against P. aeruginosa biofilms. Comparative study among the selected tests shows similar results between CFUs counts and spectrophotometry, but there are significant differences between direct counting and McFarland test. At concentration of 1.61%, the correlation factors were 0.8998 and 0.7338 when comparing CFU direct counting with microplate absorbance measurements and McFarland turbidity tests, respectively. These results show that measurement on microplates is an effective tool to evaluate the effectiveness of PERAsafe® against P. aeruginosa biofilms, being able to effectively replace the CFUs count in routine tests to determine the sterilization levels on surfaces.  相似文献   

6.
To reduce time for enumeration of viable Clostridium perfringens, fluorescence in situ hybridization in combination with filter cultivation (FISHFC) was employed. The method utilized a CLP-180 probe, based on the 16S rRNA region of C. perfringens, and FISHFC fluorescence microscopy to detect C. perfringens, but not organisms from other species. Optimal cultivation requirements for micro-colony formation were TSC medium, anaerobic conditions, 37 °C, and incubation for 6 h. Under these conditions, micro-colony diameters reached 100 μm, a size sufficient for hybridization. Enumeration of C. perfringens using the CLP-180-aided FISHFC method was realized in 9 h as compared to 3–5 days required by the conventional plate count method. Moreover, viable C. perfringens counts of food samples from the two methods were not significantly different.  相似文献   

7.
Flow cytometry (FCM) is a powerful tool for analyzing physiological characteristics of microorganisms on a single-cell basis and identifying heterogeneities within population. This work analyzed the UV-C induced damage on Escherichia coli ATCC 11229; Listeria innocua ATCC 33090 and Saccharomyces cerevisiae KE162 cells by applying flow cytometry technique. The UV-C doses, obtained by altering the exposure time and measured by the iodide-iodate chemical actinometer, ranged between 0 and 5 kJ/m2. E. coli; L. innocua and S. cerevisiae populations were quantified by plate count technique. For flow cytometry studies, cells were labeled with fluorescein diacetate (FDA) for detecting membrane integrity and esterase activity, and with propidium iodide (PI) for monitoring membrane integrity. The results showed that mechanisms of cellular damage differed according to time of exposure to ultraviolet radiation and the organism tested. E. coli and S. cerevisiae sub-populations with PI increased within the first minutes of UV-C treatment, without much change afterwards. On the contrary, FCM was used to detect the inactivation of those L. innocua sub-populations of viable microorganisms (maintaining metabolic activity) which were non-culturable due to membrane rupture and thus not detectable by viable plate count technique.  相似文献   

8.
Sourdoughs, in which Lactobacillus sanfranciscensis is the predominant bacterial species, are distinctive of some traditional Italian sweet baked products like Panettone. The direct extraction of amplifiable bacterial DNA from products subjected to heat treatment represents a valid tool to identify and trace microbial species originally present in the food matrices. Three types of protocols for the isolation and clean-up of DNA (CTAB, Wizard® DNA Clean-Up System, NucleoSpin® Food) were applied on mother, final dough and end-product samples and compared through the determination of the maximum amplifiable dilution by a PCR reaction targeting two fragments (1460 and 153 bp long) of 16S rDNA region of Lb. sanfranciscensis. CTAB extracting protocol was revealed to be the best for isolating DNA. In dough samples the amplification with the 153 bp fragment showed signals at concentration levels that are comparable with the values obtained from the plate counts, and two log cycles higher than those found with the amplification targeting the 1460 bp fragment. In the cooked samples only the 153 bp amplicon was detected, indicating that oven cooking degrades DNA into small fragments.  相似文献   

9.
Potential effects of the fat content of frankfurters on the gastrointestinal survival of Listeria monocytogenes were investigated. At various stages of storage (7 °C, up to 55 days), inoculated frankfurters of low (4.5%) and high (32.5%) fat content were exposed to a dynamic gastrointestinal model (37 °C) and L. monocytogenes counts were determined at intervals during exposure in each gastrointestinal compartment (gastric, GC; intestinal, IC). Bacterial survival curves in each compartment were fitted with the Baranyi and Roberts mathematical model. L. monocytogenes populations on low- and high-fat frankfurters exceeded 8.0 log CFU/g at 39 and 55 days of storage, respectively. Major declines in populations occurred after 60 min on low-fat frankfurters in the GC, with reductions of 2.6 to >7.2 log CFU/g at 120 min on days 1 and 39 of storage, respectively. L. monocytogenes reductions in high-fat frankfurters ranged from 1.6 (day-1) to 5.2 (day-55) log CFU/g. Gastric inactivation rates were 0.080–0.194 and 0.030–0.097 log CFU/g/min for low- and high-fat samples, respectively. Since gastric emptying began while the gastric pH was >5, initial counts (enumerated 30 min after ingestion) reaching the IC depended on initial contamination levels on each product, which increased during storage. Subsequent reductions during the intestinal challenge were 0.1–1.4 log CFU/g. Findings indicated protective effects of fat against gastric destruction of L. monocytogenes. However, since the effects of fat were observed mainly at later stages of gastric exposure, they did not influence numbers of viable cells reaching the IC.  相似文献   

10.
Campylobacter jejuni is an important foodborne gastrointestinal pathogen and highly sensitive to environmental stresses. Research has shown that changes in culturability, cell morphology, and viability occur when C. jejuni cells are subjected to stresses. In this study, real-time PCR, ethidium monoazide (EMA) in combination with real-time PCR (EMA-PCR), BacLight bacterial viability staining, and agar plate counting methods were used to quantitatively analyze viable, stressed, and dead C. jejuni strain 81-176. The real-time PCR assay provides highly sensitive and specific quantification of total genome copies of C. jejuni culture in different growth phases. Our results also reveal that real-time PCR can be used for direct quantification of Campylobacter genome release into Phosphate Buffered Saline (PBS) as an indicator of cell lysis. Using EMA-PCR, we obtained a dynamic range of greater than 3 logs for differentiating viable vs. dead cells. The viability and morphological characteristics of the stressed cells after one-week incubation at 25 °C, in air, and under nutrient-poor conditions were investigated. Our results indicated that, over 99% of the stressed cells were converted from the spiral to the coccoid form and became non-culturable. However, more than 96% of the coccoid cells retained their membrane integrity as suggested by both the BacLight staining and EMA-PCR analyses. Thus, to detect C. jejuni under stress conditions, conventional culturing method in conjunction with EMA-PCR or BacLight staining might be a more appropriate approach.  相似文献   

11.
《Food microbiology》1998,15(5):539-550
Production of viable but non-culturable (VNC)Campylobacter jejunicells was studied using 36 strains resuspended in microcosm water. Cells in stationary phase were resuspended at 4°C, with gentle stirring at 100 r min−1, in filtered surface water adjusted to pH 6.050. The culturability of starved cell suspensions was determined by spread plate counts, and the activity of cells by the direct viable count technique and CTC–DAPI double staining. Although mostC. jejunistrains died within a few days of suspension in microcosm water, three strains (Bf, 79 and 85) demonstrated a complete decrease of culturability within 15 days of suspension. A constant activity level was found in these three strains, even after 30 days of starvation in microcosm water; and an almost constant concomitant level of adenosine triphosphate was measured in VNC cell suspensions. Scanning electron microscopy ofC. jejuniVNC cells showed that the spiral-shaped cells of strain 85 cultures became coccoid in ageing microcosm-water cell suspensions, whereas most VNC cells of strains Bf and 79 remained spiral-shaped after 30 days of starvation.  相似文献   

12.
We evaluated the influence of ultrahigh pressure homogenization (UHPH) treatment applied to milk containing Staphylococcus aureus CECT 976 before cheese making, and the benefit of applying a further high hydrostatic pressure (HHP) treatment to cheese. The evolution of Staph. aureus counts during 30 d of storage at 8°C and the formation of staphylococcal enterotoxins were also assessed. Milk containing approximately 7.3 log10 cfu/mL of Staph. aureus was pressurized using a 2-valve UHPH machine, applying 330 and 30 MPa at the primary and the secondary homogenizing valves, respectively. Milk inlet temperatures (Tin) of 6 and 20°C were assayed. Milk was used to elaborate soft-curd cheeses (UHPH cheese), some of which were additionally submitted to 10-min HHP treatments of 400 MPa at 20°C (UHPH+HHP cheese). Counts of Staph. aureus were measured on d 1 (24 h after manufacture or immediately after HHP treatment) and after 2, 15, and 30 d of ripening at 8°C. Counts of control cheeses not pressure-treated were approximately 8.5 log10 cfu/g showing no significant decreases during storage. In cheeses made from UHPH treated milk at Tin of 6°C, counts of Staph. aureus were 5.0 ± 0.3 log10 cfu/g at d 1; they decreased significantly to 2.8 ± 0.2 log10 cfu/g on d 15, and were below the detection limit (1 log10 cfu/g) after 30 d of storage. The use of an additional HHP treatment had a synergistic effect, increasing reductions up to 7.0 ± 0.3 log10 cfu/g from d 1. However, for both UHPH and UHPH+HHP cheeses in the 6°C Tin samples, viable Staph. aureus cells were still recovered. For samples of the 20°C Tin group, complete inactivation of Staph. aureus was reached after 15 d of storage for both UHPH and UHPH+HHP cheese. Staphylococcal enterotoxins were found in controls but not in UHPH or UHPH+HHP treated samples. This study shows a new approach for significantly improving cheese safety by means of using UHPH or its combination with HHP.  相似文献   

13.
Bacillus sporothermodurans produces highly heat-resistant spores that can survive ultra-high temperature (UHT) treatment in milk. Therefore, we developed a rapid, specific and sensitive semi-nested touchdown PCR assay combined with propidium monoazide (PMA) treatment for the detection of viable B. sporothermodurans vegetative cells. The semi-nested touchdown PCR alone proved to be specific for B. sporothermodurans, and the achieved detection limit was 4 CFU/mL from bacterial culture and artificially contaminated UHT milk. This method combined with PMA treatment was shown to amplify DNA specifically from viable cells and presented a detection limit of 102 CFU/mL in UHT milk. The developed PMA-PCR assay shows applicability for the specific detection of viable cells of B. sporothermodurans from UHT milk. This method is of special significance for applications in the food industry by reducing the time required for the analysis of milk and dairy products for the presence of this microorganism.  相似文献   

14.
Two types of probiotic cheese, with free and microencapsulated bacteria, were manufactured in triplicate under the same conditions. The number of viable cells during 182 days of storage in refrigerated conditions was evaluated. The number of viable cells of Lactobacillus acidophilus was reduced significantly from day 28 to day 182 of storage period in both types of cheese, but reduction in the cheese containing free cells (5.1 ± 0.67 log cfu g−1) was significantly p < 0.05 higher than the cheese containing microencapsulated cells (11.00 ± 0.58 log cfu g−1). The results showed that, microencapsulation in calcium alginate gel and resistant starch was able to increase the survival rate of L. acidophilus La5 in Iranian white brined cheese after 6 months of storage.  相似文献   

15.
The microbiological responses of two bivalves species from Tagus estuary, Venerupis pullastra (native clam) and Ruditapes philippinarum (exotic clam) were investigated during 48 h of depuration and subsequent simulated transport in semi-dry conditions at two temperatures (4 and 22 °C) until reaching 50% lethal time (LT50). Regardless of temperature and species, the maintenance of clams in water for 48 h (depuration period) did not affect LT50 during transport. R. philippinarum showed higher survival rates than V. pullastra, always reaching LT50 later, especially at 4 °C. Significant differences between clams' species were found in almost all microbiological parameters. This can be related with clams' biological activity and habitat environmental conditions since both clams do not coexist in Tagus estuary. Depuration was efficient to reduce the bacterial load, particularly Escherichia coli, but not efficient to remove Vibrio spp. In both species, the growth of Vibrio spp. was inhibited at 4 °C, whereas exponential growth occurred at 22 °C. Total viable counts significantly increased in most treatments, while E. coli counts significantly decreased to undetected levels, except for non-depurated R. philippinarum simulated transported at 4 °C. Thus, this study highlights the importance of clams depuration for at least 24 h in polluted estuarine areas, followed by transport at low temperatures (4 °C).  相似文献   

16.
Bdellovibrio-and-like organisms (BALOs) are a group of highly motile delta-proteobacteria that prey on other gram-negative bacteria. However, nothing is known of the application potential of marine BALOs in safeguarding seafood safety. Here, biological characterization of two marine BALOs strains and their application in the elimination of Vibrio parahaemolyticus in oyster (Crassostrea ariakensis) at the laboratory scale were investigated.BALOs strains BDH12 and BDHSH06 were isolated from sediment of Daya bay in Shenzhen of China, with Shewanella putrefaciens strain 12 and V. parahaemolyticus strain SH06 as preys, respectively, when using double layer agar technique. They were identified as BALOs morphologically by transmission electron microscopy, while partial 16S rDNA sequencing analysis revealed that they showed no close relationships with members of the known genera Bdellovibrio, Bacteriolyticum, Bacteriovorax, or Peredibacter.Biological characterizations revealed that both strains had the optimal pH, salinity and temperature at 7.2, 3% and 30 °C, correspondingly. They could not utilize autoclaved, dead cells as hosts. Prey range analysis revealed that individually, BDH12 and BDHSH06 lysed 82.5% (47 strains) and 84.2% (48 strains) of the total 57 preys tested respectively. In combination, they lysed 98.2% (56 of 57) strains. All strains of V. parahaemolyticus, Vibrio cholerae and Vibrio alginolyticus tested could be lysed by both strains.A 7-day laboratory-scale V. parahaemolyticus elimination experiment in oyster showed that in the control, the cell counts of total vibrios and V. parahaemolyticus strain Vp plus in water and in oyster intestines were on the rise, whereas in the BALOs treated groups, their numbers were down from 8.09 ± 0.05 log CFU/ml and 8.02 ± 0.04 log CFU/ml to 2.39 ± 0.01 log CFU/ml and 2.33 ± 0.01 log CFU/ml, respectively. The same patterns could also be observed in oyster intestines. Results of this study indicate the feasibility of using BALOs to biologically control or even eliminate V. parahaemolyticus in seafood oyster.  相似文献   

17.
High hydrostatic pressure (HHP) treatments (singly or in combination with natural antimicrobials) were tested for stabilization of Manzanilla Aloreña seasoned olives stored at 25 °C. HHP (5 min) was highly effective on yeast populations at 300 MPa or higher. No viable yeasts were detected in samples treated at 400 MPa for up to three months. Low levels of endospore-forming bacteria were always detected after HHP treatments. Addition of nisin to the brines reduced bacterial counts by 1.4 log cycles but it had no effect on yeasts when tested singly or in combination with HHP treatment (400 MPa, 5 min). Thyme oil had almost no effect on yeast concentrations, but rosemary oil reduced yeast viable counts progressively during storage. Essential oils in combination with HHP (400 MPa, 5 min) significantly (p < 0.05) reduced the concentrations of aerobic mesophilic bacteria. Low-salt brined olives purged with N2 or supplemented with ascorbic acid and then pressurized for 5 min at 450 or 550 MPa were preserved for up to 5 months without spoilage, suggesting that the NaCl content in brines of packed Manzanilla Aloreña table olives could be reduced considerably by application of HHP as a stabilization treatment.  相似文献   

18.
Cell survival of Lactobacillus rhamnosus GG entrapped in gelled whey protein isolate (WPI) micro-beads was elucidated relative to cells suspended in native WPI and free-cell controls during ex vivo porcine gastro-intestinal incubation. Probiotic gastric tolerance was investigated as a function of pH (2.0-3.4) and time with subsequent intestinal incubation (pH 7.2). Free cells showed no survival after 30 min ex vivo stomach incubation (≤pH 3.4), while native WPI enhanced survival by 5.7 ± 0.1, 5.1 ± 0.2 and 2.2 ± 0.2 log10 cfu mL−1 following 180 min incubation at pH 3.4, 2.4 and 2.0, respectively. Protein micro-beads augmented ex vivo probiotic acid resistance (8.9 ± 0.1 log10 cfu mL−1) and demonstrated significant micro-bead adsorption capacity coupled with micro-bead digestion and controlled release of viable, functional probiotics within 30 min intestinal incubation. This technology potentially envisions whey protein micro-beads as efficacious entrapment matrices and binding vehicles for delivery of bioactive ingredients.  相似文献   

19.
This work aimed to investigate the influence of physicochemical properties and prior mode of growth (planktonic or sessile culture) on attachment of 13 Campylobacter jejuni strains and 5 Campylobacter coli strains isolated from chicken samples to three abiotic surfaces: stainless steel, glass and polyurethane. Water contact angle and zeta potential measurements indicated that the strains varied with respect to surface hydrophobicity (17.6 ± 1.5 to 53.0 ± 2.3°) and surface charge (−3.3 ± 0.4 to −15.1 ± 0.5 mV). Individual strains had different attachment abilities to stainless steel and glass (3.79 ± 0.16 to 5.45 ± 0.08 log cell cm−2) but did not attach to polyurethane, with one exception. Attachment of Campylobacter to abiotic surfaces significantly correlated with cell surface hydrophobicity (P ≤ 0.007), but not with surface charge (P ≥ 0.507). Cells grown as planktonic and sessile culture generally differed significantly from each other with respect to hydrophobicity and attachment (P < 0.05), but not with respect to surface charge (P > 0.05). Principal component analysis (PCA) clustered strains into three groups (planktonic culture) and two groups (sessile culture) representing those with similar hydrophobicity and attachment. Of the four highly hydrophobic and adherent strains, three were C. coli suggesting that isolates with greater hydrophobicity and adherence may occur more frequently among C. coli than C. jejuni strains although this requires further investigation using a larger number of strains. Assignment of pulsed-field gel electrophoresis profiles to PCA groups using Jackknife analysis revealed no overall relationship between bacterial genotypes and bacterial attachment. No relationship between serotype distribution and bacterial attachment was apparent in this study.  相似文献   

20.
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