Temperature in agar plates and its influence on the results of quantitative microbiological food analyses.

The numbers of colony forming units (cfu) of some strains of Enterobacteriaceae growing in Violet Red bile agar at 44 degrees C can vary considerably depending on incubation conditions and location of an individual agar plate in a stack. The reason is that the heating-up rates of the agar in plates incubated at elevated temperatures are slower if the incubators do not have an air circulator and/or if the plates are located in the centre of a stack. Variability in agar temperatures among different plates after 24 h of incubation at 44 degrees C was higher if plates were placed in an incubator with an air circulator than in an incubator without one. This effect could be avoided if the plates were incubated enclosed in a plastic bag.     

Removal of oxide nanoparticles in a model wastewater treatment plant: influence of agglomeration and surfactants on clearing efficiency

The rapidly increasing production of engineered nanoparticles has created a demand for particle removal from industrial and communal wastewater streams. Efficient removal is particularly important in view of increasing long-term persistence and evidence for considerable ecotoxicity of specific nanoparticles. The present work investigates the use of a model wastewater treatment plant for removal of oxide nanoparticles. While a majority of the nanoparticles could be captured through adhesion to clearing sludge, a significant fraction of the engineered nanoparticles escaped the wastewater plant's clearing system, and up to 6 wt % of the model compound cerium oxide was found in the exit stream of the model plant. Our study demonstrates a significant influence of surface charge and the addition of dispersion stabilizing surfactants as routinely used in the preparation of nanoparticle derived products. A detailed investigation on the agglomeration of oxide nanoparticles in wastewater streams revealed a high stabilization of the particles against clearance (adsorption on the bacteria from the sludge). This unexpected finding suggests a need to investigate nanoparticle clearance in more detail and demonstrates the complex interactions between dissolved species and the nanoparticles within the continuously changing environment of the clearing sludge.     

Effect of surface properties of different food contact materials on the efficiency of quaternary ammonium compounds residue recovery and persistence

Residues of quaternary ammonium compounds (QACs) remaining after sanitising were evaluated for a number of materials used in food plants. The residues were collected by swabs and measured using a spectrophotometric method. Surface topography and energy affected the QACs recovery. Highest percentage of QACs recovery was achieved for the tile material (102.2%) which had the most hydrophilic properties and least irregularities in surface topology, followed by stainless steel (82.1%). Meanwhile, the lowest recovery occurred in PVC (42.1%) and resin (44.3%) that exhibited hydrophobic characteristics and abrupt changes in height profile in a given surface area. Monitoring of QACs residues deposited on the surfaces after 7 days showed that the recovery of QACs in PVC and resin reduced significantly (P < 0.05), supposing that QACs might be degraded or interacted with the materials. However, no significant changes in residue recovery were observed for tiles and stainless steel surfaces after 7 days.     

Experimental comparison of excision and swabbing microbiological sampling methods for carcasses

Bovine sides, ovine carcasses, and porcine carcasses were individually inoculated by dipping in various suspensions of a marker organism (Escherichia coli K-12 or Pseudomonas fluorescens), alone or in combination with two meat-derived bacterial strains, and were sampled by two standard methods: cotton wet-dry swabbing and excision. The samples were examined for bacterial counts on plate count agar (PCA plate counts) and on violet red brilliant green agar (VRBGA plate counts) by standard International Organization for Standardization methods. Average bacterial recoveries by swabbing, expressed as a percentage of the appropriate recoveries achieved by excision, varied widely (2 to 100%). Several factors that potentially contributed to relatively low and highly variable bacterial recoveries obtained by swabbing were investigated in separate experiments. Neither the difference in size of the swabbed area (10, 50, or 100 cm2 on beef carcasses) nor the difference in time of swabbing (20 or 60 min after inoculation of pig carcasses) had a significant effect on the swabbing recoveries of the marker organism used. In an experiment with swabs preinoculated with the marker organism and then used for carcass swabbing, on average, 12% of total bacterial load was transferred inversely (i.e., from the swab to the carcass during the standard swabbing procedure). In another experiment, on average, 14% of total bacterial load was not released from the swab into the diluent during standard swab homogenization. Use of custom-made swabs with abrasive butts, around which metal pieces of pan scourers were wound, markedly increased PCA plate count recoveries from noninoculated lamb carcasses at commercial abattoirs compared with cotton swabs. In spite of the observed inferiority of the cotton wet-dry swabbing method compared with the excision method for bacterial recovery, the former is clearly preferred by the meat industry because it does not damage the carcass. Therefore, further large-scale evaluation of the two carcass sampling methods has been undertaken under commercial conditions and reported separately.     

Effectiveness of a dry kitchen system on controlling the microbiological safety of food and contact surfaces

The aim of this study was to examine the effect of dry and wet kitchens on the microbial quality of food and facility. Samples were 3 foods items and 4 surface samples taken from 2 dry and 4 wet kitchens in Korea. Total mesophilic count (TMC), coliforms, Escherichia coli, Staphylococcus aureus, Salmonella sp. were analyzed. The TMC levels of the food were within 5.97 log CFU/g, but E. coli were detected from 1 wet kitchen. The compatibility rates of the surface samples in the dry kitchen were significantly higher than those in the wet kitchen for TMC, coliforms and E. coli. As the results of the growth rate of TMC and coliforms under simulation conditions, the dry kitchen samples showed longer lag time and slower specific growth rate than wet kitchen samples. In conclusion, dry kitchen floor is a more effective way for ensuring the microbiological safety of food.     

Commercialization conditions and practices influence the microbiological quality of mineral waters

The objective of the present study was to determine the microbiological quality of bottled mineral water marketed in commercial establishments and by street vendors and to evaluate the influence of the storage and maintenance conditions on the microbiological quality of the product. Ten samples from the same batches of five different brands of water were analyzed, for a total of 50 samples. Of the five brands analyzed, only one (brand A), when collected in a commercial establishment, complied with the legal Brazilian standards for mineral water with respect to the presence of total coliforms, fecal coliforms, and Pseudomonas aeruginosa. The remaining samples failed to comply with these microbiological standards for at least one of the parameters evaluated. The water samples obtained from street vendors were inferior in microbiological quality to samples from the same batch that were obtained from commercial establishments.     

The influence of extrinsic factors on the microbiological spoilage pattern of ground beef

Two hundred and thirty four samples of raw minced beef were subjected to storage at 0 and 7 degrees C over a period of 17 days. The samples were subjected to four different treatments where the controls (Treat 1) were aerobically packed. The vacuum-packed samples (Treats 2 and 3) differed only by the addition of 0.5% L-(+)-ascorbic acid to the Treat 3 samples. Treat 4 represented aerobically packed samples to which a commercial 'colour retainer' was added. The microbiological results showed clearly that temperature control around 0 degree C was the central element in achieving shelf life extension of raw minced beef. Vacuum packaging and additive treatments enhanced the effect of low storage temperatures. Identification of 128 psychrotrophic spoilage isolates revealed a predominance of Gram-negative bacteria (63%), most of which were classified as Pseudomonas spp. (72%), the rest being Enterobacteriaceae. Among Gram-positive isolates, lactobacilli and yeasts predominated (45% and 28%, respectively). Commonly observed oxygen relationships were not found in this study. Pseudomonads proliferated even in vacuum packaged samples throughout the entire storage period, whilst lactic acid bacteria were also found in high numbers in aerobically packaged samples. Identification of 71 isolates of lactic acid bacteria revealed a strong predominance of Lactobacillus saké (34%), followed by L. curvatus (23%), L. bavaricus (21%) and L. alimentarius (10%). Significant numbers of lactobacilli were isolated from samples in all treatment groups, including the aerobically packed categories.     

Practitioner influence on contact lens prescribing in the UK

Contact lenses are mainly fitted by registered optometrists and contact lens opticians in the UK. Data we have gathered from annual contact lens fitting surveys over the past 12 years indicate that, on average, registered optometrists and contact lens opticians undertake 3.2 and 7.1 contact lens fits per week (p < 0.0001). More experienced practitioners tend to fit older patients. Practitioners fitting more lenses per year tend to fit a higher proportion of soft lenses. Contact lens opticians tend to fit a higher proportion of patients with planned replacement and daily disposable lenses compared with optometrists.     

魔芋胶、琼脂及他拉胶对软冰淇淋品质影响研究

对反应软冰淇淋品质的各指标进行测试,以考察魔芋胶、琼脂、他拉胶对软冰淇淋品质的影响.结果表明,在没有乳化剂存在情况下,三种稳定剂相比,琼脂对软冰淇淋浆料黏度和热稳定性基本无影响,对膨胀率、抗溶性和硬度起降低作用,但相对其他2种稳定剂而言抗融性最好,硬度最大;魔芋胶对软冰淇淋浆黏度起增大作用,提高膨胀率,但硬度和抗融性最差;他拉胶对冰淇淋浆料黏度的影响随添加量的增加缓慢增大,对膨胀率、硬度和抗溶性均由降低作用,但对各指标的影响程度稍好于魔芋胶.     

Sorption of ionic surfactants to estuarine sediment and their influence on the sequestration of phenanthrene

The sorption of an anionic surfactant (sodium dodecyl sulfate; SDS) and a cationic surfactant (hexadecyl trimethylammonium bromide; HDTMA) to estuarine sediment has been studied in river water and seawater. Sorption isotherms for SDS were essentially linear in both waters, suggesting a nonspecific, hydrophobic interaction between the SDS tail and particle surface. Sorption of HDTMA was considerably greater, more nonlinear, and more sensitive to water composition. These observations were attributed to a combination of both electrostatic and hydrophobic interactions between the surfactant and particle surface, the formation of admicelles, and salinity-induced structural alteration of the hydrophobic tail of the HDTMA molecule. Presence of SDS caused a reduction in the sorption of phenanthrene to estuarine sediment because of the competitive effects of the surfactant tail for hydrophobic sorption sites on the particle surface. Conversely, the presence of HDTMA caused significant enhancement in phenanthrene sequestration because of head-on sorption of surfactant molecules and a resulting, more hydrophobic particle surface. The most persistent feature of our results was an inverse dependence of unit sorption on particle concentration, and an empirical algorithm defining the effect was used to calculate the sediment-water fractionation of realistic concentrations of reactants in the estuarine water column. The results of these calculations, and the more general findings of this study, significantly improve our understanding of both the transport and fate of ionic surfactants in the estuarine environment, and the effects that these surfactants have on the partitioning of hydrophobic organic micropollutants.     

Evaluation of gamma rays influence on some biochemical and microbiological aspects in black truffles

The effects of two gamma-ray doses (1.5 kGy and 2.0 kGy) on some biochemical aspects and on the microbiological profile of black truffles was monitored, immediately after treatment and after 30 days of storage at 4 °C. Electrophoretic and chromatographic analyses of proteins and peptides, just like monitoring of polyphenol content, peroxides formation and microbial profile, allowed for the first time a better understanding of the mechanisms responsible for biochemical alterations and bacterial pattern in black truffles during their storage. Treatment at 1.5 kGy appeared to better preserve the characteristics of the fresh product. In 2.0 kGy-samples, the protein profile was characterised by a 20 kDa-polypeptide, which could be considered as an useful marker of the irradiation treatment and of the storage time of the product. MALDI-TOF mass spectrometry analysis did not permit a correct identification from tryptic peptides in databases, although the nano-ES/MS/MS analyses performed on the 10 kDa tryptic digest peptides showed an amino acidic sequence entirely contained in a protein of filamentous fungus Neurospora crassa.     

The influence of phytosterols on the encapsulation efficiency of cholesterol liposomes

Lecithin vesicles, prepared by the dehydration‐rehydration (DR) method, were used to encapsulate bovine serum albumin (BSA). These were used to test the feasibility of making liposomes containing phytosterols, such as β‐sitosterol and stigmasterol, as a substitute for cholesterol. Their physicochemical properties such as encapsulation efficiency (EE), stabilities of storage, pH, and oxidization were investigated to assess substitutability. Liposomes prepared with phytosterols or cholesterol exhibited higher EE for BSA than those prepared without the addition of sterols. In addition, the EE of liposomes was increased with repeating a DR cycle five times. Liposomes at pH 6 or 7 were most stable, irrespective of phytosterols or cholesterol being added. Liposomes stored at 4 °C had a higher residual percentage than those stored at ?20 °C or room temperature. The addition of sterols to liposomes was effective in decreasing thiobarbituric acid‐reactive substances (TBARS) during storage periods; the antioxidation was more marked when α‐tocopherol was added to liposomes. The results indicate that replacing cholesterol with phytosterols in preparing liposomes is feasible and recommended.     

Comparison of sponging and excising as sampling procedures for microbiological analysis of fresh beef-carcass tissue

Sponging and excising were evaluated as sampling procedures for microbiological analysis of beef-carcass tissue. Brisket tissue portions (10 x 10 cm) were inoculated with 2 ml of an Escherichia coli ATCC 25922 cell suspension (3 x 10(8)CFU/ ml). After 30 min, the portions were sampled by excising (EX) or swabbing (SP) with a sterile sponge and were analyzed for aerobic plate counts on tryptic soy agar and for total coliform counts and E. coli counts on Petrifilm E. coli count plates. Another set of inoculated samples was analyzed after being spray washed, in sequence, with water (6 s, 35 degrees C, 3.4 bar), acetic acid (2%, 6 s, 35 degrees C, 2.1 bar), water (20 s, 42 degrees C, 20.7 bar), and acetic acid (2%, 6 s, 35 degrees C, 2.1 bar). Additional samples were sampled for analysis after chilling at 7 degrees C for 24 h. Bacterial counts recovered were influenced (P < or = 0.05) by procedure of sampling (EX versus SP), time of sampling (0.5 versus 24 h), and by their interactions. Counts recovered 0.5 h after inoculation from unwashed or spray-washed samples were similar between the two sampling procedures (EX and SP). However, counts recovered after 24 h of sample storage were significantly (P < or = 0.05) lower for the SP compared with the EX procedure. The results indicated that as the carcass tissue was stored, recovery of bacteria by SP was less efficient than was recovery by EX.     

Time-dependent competitive adsorption of milk proteins and surfactants in oil-in-water emulsions

Competitive adsorption of pure milk proteins and non-ionic surfactants has been studied in model oil-in-water emulsions (4 g kg^?1 β-lactoglobulin or β-casein, 200 g kg^?1 n-hexadecane) as a function of the age of the adsorbed protein layer at the oil-water interface. With β-lactoglobulin-stabilised emulsions containing oil-soluble surfactant C₁₂ E₂ (diethylene glycol n-dodecyl ether), there is found to be a steadily increasing amount of protein associated with the emulsion droplets over a few hours following emulsification. Addition of water-soluble surfactant Tween 20 (polyoxyethylene (20) sorbitan monolaurate) to a β-lactoglobulin-stabilised emulsion (with or without C₁₂E₂) leads to less protein displacement if the emulsion is aged prior to addition of Tween 20. Moderate additions of C₁₂E₂ or Tween 20 produce no time dependence in the competitive adsorption in β-casein-stabilised emulsions, although some time dependence is observed when C₁₂E₂ and a high concentration of Tween 20 are present together. Crystallisation of the oil phase in β-casein-stabilised emulsions at pH 7 leads to a lowering of the measured protein surface concentration, especially in the presence of C₁₂E₂ and a reduction in the surfactant to protein molar ratio required for complete protein displacement by water-soluble surfactant (Tween 20 or octaethylene glycol n-dodecyl ether). Under more acidic conditions of pH 5 or pH 3, the surface coverage and ease of displacement of β-lactoglobulin at the surface of liquid emulsion droplets is substantially different from that under neutral pH conditions.     

An assessment of sampling methods and microbiological hygiene indicators for process verification in poultry slaughterhouses

Studies to determine the appropriateness of the use of populations of indicator bacteria on poultry carcasses for process verification were undertaken in commercial slaughterhouses. Samples were collected from neck skin by excision or from whole carcass rinses and were examined for a range of presumptive process hygiene indicator bacteria. Coefficients of variation were calculated for each bacterial indicator and were significantly lower in excised samples, indicating more reproducible bacterial recovery by this sampling method. Total viable counts of aerobic bacteria, Enterobacteriaceae, and Pseudomonas in samples collected by excision had the lowest coefficients of variation when compared with other indicators and were therefore used for further study. The uncertainties associated with the quantification of each bacterial indicator were calculated and were lowest overall for total viable counts of aerobic bacteria. In general, uncertainty was higher for lower bacterial numbers. Results of microbiological testing on pooled excised neck skin samples were not significantly different from the mean of individually analyzed samples. Bacterial numbers increased by 1 log unit when cultures were stored under chilled conditions typical of those used for transporting samples to external laboratories, but the increases were not significant for Pseudomonas and aerobic bacteria when storage time was less than 17 h. Weak relationships were identified between bacterial indicator numbers and duration of processing, although cleanliness of the processing environment diminished visibly during this time. In the plants visited for this study, there was a poor relationship between presumptive bacterial indicator numbers and process hygiene. Consequently, bacterial analyses for process verification purposes may be of limited value.     

An in vitro system for the comparison of excision and wet-dry swabbing for microbiological sampling of beef carcasses

An in vitro system for the comparison of wet-dry swabbing and surface tissue excision was developed to ascertain whether the commonly accepted statement of the advantage (in terms of bacterial recovery) of the tissue excision method is also legitimate when different kinds of bacteria are used. A total of 1,770 sections (2.5 by 10 cm) of bovine skin were individually inoculated on the subcutaneous fat side by spreading various suspensions of marker organisms (nalidixic acid-resistant Escherichia coli, vancomycin-resistant Enterococcus faecalis, and methicillin-resistant Staphylococcus aureus) at different concentrations and sampled by two standard methods: cotton wet-dry swabbing and excision. Most counts from cuts sampled by excision were significantly (P < 0.05) higher than the wet-dry swabs; however, no differences were observed between the control and the sampling method when sections were inoculated with bacterial solutions at a concentration of 10(3) CFU/ml and sampled by excision. For sections inoculated with bacterial solutions at a concentration of 10(3) CFU/ml, counts given as log CFU/25 cm2 ranged from 1.97 (S. aureus sampled by wet-dry swab) to 3.06 (S. aureus sampled by excision). For sections inoculated at a concentration of 10(4), counts given as log CFU/25 cm(2) ranged from 2.15 (E. faecalis sampled by wet-dry swab) to 3.19 (S. aureus sampled by excision). For sections inoculated at 10(5), counts given as log CFU/25 cm(2) ranged from 2.94 (E. faecalis, wet-dry swab) to 3.98 (S. aureus, excision), and for sections inoculated at 106, counts given as log CFU/25 cm(2) ranged from 3.53 (E. coli, wet-dry swab) to 4.69 (S. aureus, excision). The proposed system, which enabled a considerable amount of samples to be analyzed under controlled experimental conditions and a large number of data to be generated in a short time, demonstrated among the tested microorganisms that whereas the excision method recovered the highest number of bacteria, control means were always (with the exception of an inoculum of 10(3)/ml) significantly higher than means from either of the sampling methods. Our results indicate that particular attention should be paid to the diverse microflora that can contaminate carcasses in a given slaughterhouse and that it is not appropriate to generalize by saying that the destructive method is the reference technique for the bacteriological sampling of carcasses in slaughterhouses, especially when the contamination is higher than 10(3) CFU/25 cm(2).     

The influence of ripening temperature and sampling site on the lipolysis in Reggianito Argentino cheese

The effect of ripening temperature elevation and sampling site on lipolysis in Reggianito Argentino cheese was evaluated. Cheeses ripened at 12 or 18 °C and 85% relative humidity were assayed at 2, 4 and 6 months at two sampling zones (central and external). Samples were analysed to determine physicochemical parameters and the concentration of nine free fatty acids (FFAs) (C_6:0–C_18:2). Myristic, palmitic, stearic and oleic acids were found in higher concentration. While ripening time and temperature significantly affected the concentrations of the nine FFAs analysed, sampling zone significantly affected only two FFAs. Ripening temperature increased the lipolytic process, but it seems to have no effect on the pathways of lipolysis in Reggianito Argentino cheese.     

The influence of ripening temperature and sampling site on the proteolysis in Reggianito Argentino cheese

The effects of elevated ripening temperature and sampling site on proteolysis in Reggianito Argentino cheese were evaluated. Cheeses ripened at 12 or 18 °C and 85% relative humidity were analysed at 2, 4 and 6 months in 2 sampling zones (central and external). Samples were analysed to determine the physicochemical and proteolysis parameters through the urea-PAGE of the urea-soluble fraction, RP-HPLC analysis of the water-soluble fraction at pH 4.6, and the free amino acid analysis. Proteolysis was significantly affected by ripening temperature and sampling site. Urea-PAGE analysis showed that elevated temperature increased the degradation of α_s1- and β-casein. The degradation of α_s1-casein was larger in the central zone than in the external one, while β-casein degradation was similar in both zones. The majority peaks detected by RP-HPLC of the water-soluble fraction at pH 4.6 and free amino acids were significantly affected by ripening temperature and sampling site. Glu, His, Val, Leu, and Lys had the higher concentrations. Principal component analysis showed useful groupings when results from chromatograms were studied. In conclusion, the results obtained not only are useful to characterise the ripening of an Argentinean hard cheese, but also to evaluate the effect of an increase of ripening temperature on Reggianito Argentino cheese proteolysis.