ALBERT

Notes: Live cells of E. coli O157:H7 were labeled by 4′,6-diamidino-2-phenylindole (DAPI) in buffers of different pH. The extent of labeling was relatively insensitive to pH in the range of 6.5 to 9.5. The fluorescence intensity of ± 104 DAPI-labeled bacteria per mL in optical cuvettes could be detected by a luminescence spectrometer. With a fluorescence microplate reader attachment, less than 103 of labeled bacteria could be measured. DAPI-labeling inhibited the growth and respiratory activities of the bacteria. The addition of 0.5 to 6 mM concentrations of ATP induced a substantial increase in the fluorescence of labeled bacteria. Maximal enhancement by ATP was observed from bacteria still maintaining low levels of physiological activities. The enhancement favored more alkaline media with pH greater than 9. A replacement of ATP with ADP or AMP diminished the extent of enhancement. Other triphosphate nucleotides did not enhance fluorescence of DAPI-labeled bacteria. Comparable ATP enhancements were also observed with Pseudomonas alcaligenes and Shewanella putrefaciens. Solubilization/destruction of cell membranes of labeled bacteria by detergents essentially eliminated the ATP enhancement. Absorption and fluorescence spectroscopic measurements indicated that ATP could interact with free and bound DAPI. These results suggest that observed ATP enhancement in fluorescence intensity of DAPI labels in intact cells may be applied to increase the sensitivity of microorganism detection.

Notes: Fungal biota, with special reference to the genus Penicillium, was studied in 52 samples of commercial cheeses (10 fresh, 17 semiripened and 25 ripened) made from different types of milk (cow, ewe, goat and mixed) produced in southern Spain. In 41 of the total of cheeses analyzed (79%) molds were isolated. Penicillium was identified in 63% of the samples, Mucor spp. in 27%, Geotrichum candidum in 17% and Cladosporium herbarum in 10%; eleven other fungal genera were detected ranging from 2 to 4%. Thirty-five species of Penicillium were analyzed with the following distribution: 7 in fresh cheese, 16 in semiripened cheese and 30 in ripened cheese. The incidence of Penicillium spp. was also greater in the cheeses with a higher degree of ripeness, i.e. 20% in fresh cheese, 71% in the semiripened and 76% in the ripened cheese.

Notes: Challenge studies were carried out on raw, cooked, and sterilized surimi nuggets, inoculated with 104spores/g of C. botulinum type E spores. All products were packaged in air and air with an Ageless SS oxygen absorbent and stored at 4, 12 and 25C. Toxin was not detected in any raw product throughout storage (28 days). The absence of toxigenesis was attributed to the low pH (4.1–4.3) due mainly to the growth of lactic acid bacteria (107CFU/g). Toxin was also not detected in any cooked product after 28 days. Product pH did not decrease as previously (due to the absence of LAB), but counts of C. botulinum still decreased throughout storage. In sterile nuggets, C. botulinum counts increased to 106 cfu/g at both 12 and 25C, respectively, by 28 days. Lactic acid bacteria and Bacillus spp.were not detected throughout the 28 days storage period. Toxin was detected by days 28 and 14 at 12 and 25C, respectively, and toxigenesis preceded spoilage. The absence of toxin in cooked nuggets was attributed to the anti-botulinal role by Bacillus species, the predominant spoilage bacteria in cooked nuggets.

Notes: Packages containing chubs of summer sausage were inoculated with about 108 cfu/mL of a three-strain mixture of Listeria monocytogenes and vacuum sealed. The fate of the pathogen was then monitored after pasteurization at 150F (66C), 170F (77C), 190F (88C) and 21 OF (99C) for 0 to 240 s. Pathogen numbers were reduced by about 3 log10 cfu per gram within 30, 60, or 90s at 21 OF (99C), 190F (88C), or 170F (77C), respectively, whereas numbers were reduced by 〈2.0 log10 cfu per gram after 240 s of heating at 150F (66C). The calculated D values were 2.08 min at 150F (66C), 0.84 min at 170F (77C), 0.37 min at 190F (88C), and 0.28 min at 21 OF (99C). These results establish the feasibility of using pasteurization to control L. monocytogenes in packaged summer sausage.

Notes: Book review in this ArticleWORLD HEALTH STATISTICS QUARTERLY DALLAS G. HOOVER

Notes: Decimal reduction times (D-values) for Saccharomyces cerevisiae ascospores inoculated into pasteurized orang juice ranged from 4 to 76 s at pressures between 500 and 350 MPa. At the same pressures, D-values of S. cerevisiae vegetative cells ranged from 1 to 38 s while those for the native microflora in nonpasteurized Hamlin orange juice were between 3 and 74 s. Corresponding z-values were 123, 106 and 103 MPa for ascospores, vegetative cells and native microflora, respectively. Native microorganisms that survived high pressure treatments included yeasts, gram-positive and gram-negative bacilli. Pectinmethylesterase activity in nonpasteurized Hamlin orange juice was reduced to 5% of initial activity after 30 s at 900 MPa.

Notes: The objective of this study was to investigate awareness and attitude regarding food-related hygienic practices in the home. For this purpose and within a sanitary education program in pricary schools in a town of Central Italy, a questionnaire was used to collect information from parents of pupils. The questionnaire included questions in four major areas: personal hygiene and cleaning up procedures; meal preparation; food storage; and knowledge of key terms and concepts pertaining to food safety. Analysis of 183 questionnaires showed either risk of cross contamination, improper thawing of food, or inadequate storing and reheating of cooked foods during home food preparation and storage practice. Particularly, 73% of respondents thawed large pieces of frozen food at room temperature; 89% did not reheat cooked food after it had been stored; 75% stored raw meat and poultry in the upper shelves of refrigerators. Precise information on the lack of food safety practices will facilitate the development of proper consumer education programs.

Notes: Microbiological studies on a popular maize product (kwoka) with or without a soybean supplement (20 or 30%) were carried out during production and storage at 26–32C. Soybean -supplementedproduct had greater microbial diversity and higher populations than the unsupplemented control. The diversity was most evident at the slurry stage (blended mixture before steaming) which contained ten different microbial genera including Aspergillus, Enterobacter, Bacillus, Lactobacillus, Micrococcus and Pediococcus. A dramatic decrease occurred after steaming which killed most of the molds and Gram-negative bacteria. However, within 1 day of storage, a sharp increase was observed in the microbial population of all samples and maximum load occurred in 20% soybean supplemented “kwoka” at the end of storage. The microbiota became less diverse with storage and was dominated by Bacillus, Lactobacillus and Micrococcus. packaging of ‘kwoka’ in traditional leaves was microbiologically inferior to polyethylene packaging. Changes in the acidity of 30% supplemented ‘kwoka’ were less dramatic compared to the other products. The lactics were primarily responsible for the spoilage of the products (especially the 20% supplemented) after approximately 3 days of storage at tropical ambient temperature of26–32C.; Accepted for Publication August 8, 1997

Notes: Chicken, lamb and buffalo meat were subjected to low-dose gamma irradiation (2.5 kGy) and stored at 0–3C. Lipid peroxidation in terms of thiobarbituric acid (TBA) number and carbonyl content were monitored during storage. While irradiated meat showed slight increase in TBA number and carbonyl content on storage as compared to nonirradiated meat, this did not affect the sensory qualities of meat. Free fatty acid content decreased markedly on irradiation. Irradiated meats were microbiologically safe and sensorily acceptable up to 4 weeks in the nonfrozen state (0–3C) while nonirradiated meat had a shelf-life of less than 2 weeks.

Notes: In a nonhypothetical laboratory experiment, participants were willing to pay an average of $0.71 for the right to exchange a typical meat sandwich for a sandwich irradiated to eliminate the potential risk of foodborne bacteria. Forty-one of sixty participants (68.3%) were willing to pay some positive amount. Tobit analysis was used in order to allow for the fact that Willingness To Pay (WTP) was censored at zero. WTP was interpreted as the demand for irradiation to control foodborne disease. A positive WTP was interpreted as acceptance of irradiation. The aggregated WTP greatly exceeds the estimated direct costs of foodborne disease, and the estimated costs of irradiation for many food products. There was a positive relationship between WTP and the perceived risk of foodborne disease, and a negative relationship between WTP and years of education.

Notes: The potential of using trisodium phosphate (TSP) to reduce bacterial populations in fresh fishery products was explored since TSP has recently been approved by USDA for its usage in poultry processing to eliminate Salmonella contamination. Fresh headed shrimp and rainbow trout fillets were inoculated with L. monocytogenes before dipped in tap water, 10% TSP, or 20% TSP solutions and overwrap-packaged. Surface pH values, psychrotrophic plate counts, and L. monocytogenes counts of inoculated shrimp and trout fillets were determined after 0, 3, 6, and 9 days of storage at 4C. The TSP treatment resulted in relatively high residual surface pH values (11–12) initially in both shrimp and trout fillets. Compared to tap water dipping, TSP treatment did not significantly reduce psychrotrophic or Listeria populations in shrimp. However, the 20% TSP treatment significantly (p 〈 0.05) lowered 0-day psychrotrophic and L. monocytogenes counts of trout fillets and remained effective for 6 days during storage at 4C.

Notes: Enterohemorrhagic Escherichia coli O157:H7 (EHEC) is recognized as a frequent cause of gastroenteritis ranging from mild to severe bloody diarrhea. The Shiga-like toxin produced by EHEC can result in hemolytic uremic syndrome, now the major cause of acute kidney failure in children in the United States. Inadequately cooked beef is most commonly implicated in the transmission of EHEC, although only a small fraction of cattle appear to harbor the organism. In several studies EHEC positive herds were detected only in the summer months correlating with the occurrence of human infections. Numerous E. coli strains have been shown to enter a viable but nonculturable (VBNC) state as a result of environmental stresses, including low temperature. Using traditional plating methods and the BacLight Molecular Probe, we monitored EHEC strains incubated in river water (RW) and artificial sea water (ASW) at temperatures of 5C and 25C for their entry into the VBNC state. EHEC strains remained culturable for over 40 days in both ASW and RW incubated at 25C. In ASW, these levels were higher than a non-EHEC control. At 5C, the number of culturable EHEC cells dropped gradually in both RW and ASW. Using the BacLight Molecular Probe, we were able to demonstrate that these cells, though not culturable, were viable indicating entry into the VBNC state. Our study suggests that temperature and not salinity is the primary signal for entry into this state.

Notes: Inactivation of Escherichia coli O157:H7 by pulsed electric field, benzoic or sorbic acid, alone or in combination was investigated. When the cells of E. coli O157:H7 were suspended in 10% glycerol and treated with a single highvoltage (12.5 kv/cm) electric pulse at 25C, the count decreased by 1.1–1.6 log10 CFU/mL. Presence of benzoic and sorbic acid (1000 ppm) in the suspending medium, at pH 3.4 without electric treatment, decreased the count by 1.9–2.5 and 0.6–1.1 log, respectively. A synergistic killing effect between the high electric pulse and organic acid was observed at pH 3.4, but not at pH 6.4. When the cell suspension of E. coli O157:H7 was treated with five electric pulses in the presence of benzoic or sorbic acid at pH 3.4, the count decreased by 5.6 and 4.2 logs, respectively. Inactivation of the pathogen by combinations of electric pulse and organic acid was enhanced by an increase in temperature, field strength, and number of electric pulses, Inactivation was greater when the cells were suspended in ionic suspending media (0.1% NaCl or 5mM phosphate buffer) than in nonionic media (10% glycerol or 1% sucrose).

Notes: The prevalence of some enteric bacteria in alfalfa sprouts obtained from public markets and supermarkets in Queretaro City was determined. In addition, the antimicrobial effect of several commercial germicides was tested on alfalfa sprouts for reduction of native coliforms and inoculated Vibrio cholerae Ol or Salmonella typhi. Escherichia coli and Salmonella sp. were detected in 74% and 1.1% of 90 of samples, respectively, and no sample tested positive to V. cholerae Ol. Coliforms ranged from 7.3 to 8.5 log10 CFU/g. Treatment of alfalfa sprouts with 200 mg/L of hypochlorite, of a commercial iodophor, or of chlorine dioxide, or with 100 mg/L of Citricidal® for 5 min, reduced native coliforms only by 1–2 log10 CFU/g. Reductions of S. typhi and V. cholerae O1 with 200 mg/L of sodium hypochlorite and 100 mg/L Citricidal® were also no more than J.5 log10 CFU/g. Sprouts from seeds contaminated with V. cholerae O1 that were irrigated daily with water containing 100 mg/L chlorine dioxide, showed significantly lower V. cholerae Ol counts than seeds irrigated with tap water (p〈0.05). However, V. cholerae O1 persisted after 8 days of sprouting, when the sprouts are ready for marketing. Treatment of seeds and sprouts with antimicrobials does not appear to be effective for reducing pathogens to safe levels.

Notes: The effects and interactions of heating temperature (5–62.5C), pH (4 – 8), NaCl (0 – 6%, w/v), and sodium pyrophosphate (0 – 0.3%, w/v) on the heat resistance of a four strain mixture of Escherichia coli O157:H7 in beef gravy were examined. Thermal death times were determined using a submerged coil heating apparatus. The recovery medium was plate count agar supplemented with 1% sodium pyruvate. Decimal reduction times (D-values) were calculated by fitting a survival model to the data with a curve fitting program. The D-values were analyzed by second order response surface regression for temperature, pH, NaCl and sodium pyrophosphate levels. The four variables interacted to affect the inactivation of the pathogen. Thermal resistance of E. coli O157:H7 can be lowered by combining these intrinsic factors. A mathematical model describing the combined effect of temperature, pH, NaCl and sodium pyrophosphate levels on the thermal inactivation of E. coli O157:H7 was developed. The model can predict D-values for any combinations of temperature, pH, NaCl and sodium pyrophosphate that are within the range of those tested.

Notes: Croquette is a prepared food made with meat or fish as its basic ingredients. Since it is a perishable, heavily handled product, microbiological hazards are associated with the process. Microbiological analyses were performed throughout the commercial processing of chicken croquettes to evaluate the microbial hazards. Moderately high levels of microorganisms were observed in raw materials, but the highest plate counts were noted following extrusion of the croquettes. Pathogenic bacteria, such as Clostridium perfringens, were detected during the process. The increase in the levels of microorganisms observed after extrusion and the presence of C. perfringens were associated with the semi-solid paste called bechamel. Rapid cooling of the bechamel was critical to avoid microbial proliferation and represented an effective critical control point for the implementation of a HACCP program in the processing of croquettes.

Notes: When ozone (1.3 mM) was bubbled for 3 min in a mixture of shredded lettuce and water, counts ofmesophilic andpsychrotrophic bacteria decreased 1.4 and 1.8 log10 cfu/g, respectively. Counts of these microorganisms on lettuce, from a different batch, decreased 3.9 and 4.6 log, respectively, during 5 min of ozone treatment. Shredded lettuce was treated with gaseous ozone, or mixed with aqueous solution of ozone (1:20 w/w) with or without bubbles. For effective delivery of ozone, stirring (low and high speed), sonication or stomaching was applied during the ozonation. Washing the lettuce with water only decreased total count on shredded lettuce by 0.74–1.0 log cfu/g. When lettuce in a treatment chamber was flushed with gaseous ozone, the total count decreased 0.85 log cfu/g, but when vacuum was applied before the ozone flush, the total count decreased 0.96 log cfu/g. Bubbling ozone in water-lettuce mixture while sonicating, high-speed stirring, or before stomaching inactivated 1.4, 1.9 and 1.9 log cfu/g, respectively. In conclusion, bubbling gaseous ozone in water is the most effective ozonation method. Efficient ozone delivery to microorganisms on lettuce requires a combination of ozone bubbling and high-speed stir.

Notes: Fresh pink salmon (Oncorhynchus gorbuscha) fillets and whole Pacific herring (Clupea harengus pallasi) were stored for two weeks at 10C to determine if significant amounts of histamine were produced before the fish spoiled. Spoilage odors in salmon were moderate by day 4 and intense by day 7, while herring had detectable spoilage by day 4 and became potent by day 6. Aerobic bacterial counts increased from 4.0 × 102/g initially to 3.6 × 108/g in salmon fillets by day 14 and from 2.3 × 103/g initially to 2.7 × 107/g in whole herring by day 14. Total volatile nitrogen increased from 1.8 to 78.5 mg N/100 g in salmon and 2.2 to 23.6 mg N/100 g in herring. Histamine was not detected in salmon, while concentrations reached 54.9 ppm in herring at day 14. However, herring were considered spoiled by day 6.

Notes: This report presents results of the investigations carried out to understand the role of fat levels in fish on postirradiation recovery/survival and growth of four pathogens generally encountered in seafoods. Fish of low [golden anchoy, (Coillia dussumierij, 0.39%] as well as high fat [Indian sardine, (Sardinella longiceps), 7.1 %] content (with endogenous biota were inoculated separately with cells of Listeria monocytogenes 036, Yersinia enterocolitica 5692, Bacillus cereus and Salmonella typhimurium; and irradiated at doses of 1 and 3 kGy. The number of viable organisms for each was monitored for two weeks at 2-4C on appropriate selective media. The four pathogens studied exhibited variation in their ability to recover from radiation damage as evidenced in terms of lag period; but their recovery and subsequent growth was not influenced by the fat levels offish. No increased growth of any of the pathogens tested was evidenced in irradiated fish, when compared with nonirradiated one.

Notes: Sporulation temperature is one of the most important factors that determine the heat resistance of spores. Bacterial spores are usually more heat resistant when they are formed at higher temperatures. Sporulation temperature also influences the effect of pH of heating menstruum and the effect of different acids on spore heat resistance. Heat resistance is maximum at neutral pH and decreases with acidification, but when spores are sporulated at high temperatures the effect of acidic pH on heat resistance is lower at high heat treatment temperatures. Lactic and acetic acid decrease spore heat resistance more than other acidulants, but only when spores had been sporulated at high temperatures. These effects should be considered by canning factories of acid/acidified foods, especially in hot climate areas, where these high temperatures can be easily reached. The level of mineralization of spores obtained at different Sporulation temperatures seems to be the most important mechanism responsible for their heat resistance.

Notes: Consumer knowledge of safe handling guidelines and their reported practices assessed by a mail survey indicated consumers considered themselves informed about food safety, and most reported taking action to minimize risk from potential hazards. However, most responses were related to nutritional changes, and 20% indicated they did not know how to reduce risks from microbiological hazards. Consumers were knowledgeable about appropriate safeguards in selection and cooking specific foods, however mistakes were reported in temperature control and handling leftovers. Consumer Reports, university scientists, health professionals and science magazines were considered ihe most reliable source of food safety information. Industry advertisement appears to raise consumer concern in some areas. Educational intervention should emphasize the importance of temperature control and sanitation rather than taste to determine safety. Consumers should receive information about protective technologies like heat and irradiation pasteurization.

Notes: Results from two consumer surveys are presented. In the first, a EU FAIR (European Union Food, Agriculture and Industrial Research Programme), supported project, three-thousand consumers (500 in each of six countries, Germany, Italy, Britain, Spain, Sweden and Ireland) were surveyed in March 1997 on their attitudes to the quality and safety of three meats, beef, pork and chicken. This paper deals with the safety aspects of the survey. Many consumers in all six countries said they were eating less beef and more chicken. Overall, the Spanish and Irish appeared to be the most concerned about the safety of meat, the British were somewhat less concerned. While many were confident that the food in shops is safe, there was a sizeable minority who believed the contrary. When looking for information on the safety of meat, butchers, whether independent or in supermarkets, are the group most trusted by consumers. In terms of consumer concerns, about 60 percent of beef consumers were very concerned about hormones, BSE (Bovine Spongiform encephalopathy), antibiotics and bacteria. Pork consumers had similar levels of concern as beef consumers about bacteria, antibiotics and hormones. For chicken there was a higher level of concern about bacteria (Salmonella) (68% were very concerned) and similar levels of concern to pork and beef for the other issues. Generally, fat was the issue of least concern for all three meats. Freshness was considered to be relatively the most helpful of 7 factors used for assessing safety of meat. The country of origin and what the animal was fed were also considered to be relatively helpful. Factors considered to be relatively less helpful were price and the name of the producer. In the second study, over 1200 pre-leaving certificate Irish school children participated in autumn 1996 in a national survey assessing young people's attitudes to meat. Half of the respondents agreed with the view that eating beef means increasing the risk of getting CJD (Creutzfeldt-Jakob Disease). Those who agreed were more likely to be less frequent eaters of beef and to have reduced their meat consumption in the post-BSE period. It was found also that females were more likely to eat less meat than males and to have reduced beef consumption. The implications of some of the findings of these two surveys are discussed.

Notes: Bovine spongiform encephalopathy (BSE) belongs to a distinct group of transmissible degenerative encephalopathies (TDE) that includes scrapie in sheep and Creutzfeldt-Jakob disease (CJD) of humans. These fatal neurological diseases are caused by unconventional but uncharacterized transmissible agents that have a number of unusual properties; this includes a high degree of resistance to inactivation. The only disinfectant that appears to be completely effective against high titres of TDE infectivity is sodium hypochlorite. Exposure to 1 to 2 M sodium hydroxide has a substantial but incomplete effect, as does autoclaving at temperatures between 132 and 138C for up to an hour; however, a combination of 2 M sodium hydroxide and autoclaving, even at the more modest temperature of 121C for 30 min, appears to be completely effective. With lower titres of infectivity, less rigorous regimes may be effective. For example, although most of the rendering procedures used to manufacture meat and bone meal (MBM) throughout the European Union (EU) have been found to be incapable of inactivating BSE and scrapie agents, one method which appeared to be effective with moderate titres of BSE and scrapie agent, involved exposure to steam at 133C for 20 min. This procedure is now the only one approved within the EU for the manufacture of MBM for feeding animals, excluding ruminants; however, the UK has introduced a ban on feeding MBM to any farmed species because of the occurrence of a new variant form of CJD in the UK which appears to have been caused by the BSE agent. Although the clinical signs of scrapie in sheep are entirely attributable to neurological dysfunction, tissues such as spleen and lymph nodes are known to become infected before those of the central nervous system. In contrast, the only tissues which have been found to become infected in cattle with natural BSE are brain, spinal cord and retina. These are no longer used in animal or human foodstuff. Because scrapie agent has no known association with human disease there has been no restriction in the past on using any sheep tissues, including brain, as human food; however, it is known from experimental studies that the BSE agent can infect sheep by the oral route to cause a disease that is clinically and neurohistopathologically indistinguishable from scrapie. It is also known that the spleen becomes infected in such sheep. Sheep in the UK could theoretically have become infected with the BSE agent through the feeding of infected MBM before the feed-ban in 1988. There are now measures in place that prevent the head, spinal cord, and spleen of sheep being incorporated into foodstuff.

Notes: Considerable time, regulation and consumer concern is associated with the question of chemical residues in food. Potential residues in food span a broad range from natural contaminants (mycotoxins) through environmental contaminants (dioxins, PCBs), agrochemicals (nitrates, pesticides), veterinary drugs (anthelmintics, antibiotics), prohibited substances (hormonal growth promoters), chemicals arising in processing (nitrosamines), packaging components (plasticisers) to contaminants arising in domestic food preparation (heavy metals). The occurrence or avoidance of these chemicals as residues in food is to a greater or lesser extent under the control of the food producer, processor and consumer in the activities of growing/producing, treating, storing, processing, packaging and cooking. The fundamental question is to what extent do or can residues in food constitute a food safety hazard for the consumer. Many of these substances are regulated as to their use and as to their acceptable levels in food while others, by their nature, arise as more random occurrences and, as such, are less prone to regulation. Of particular concern to the producer, processor and retailer of food is what steps may be taken to ensure that the food supply is safe. Best practice in plant and animal husbandry by the producer, in sourcing of material and in control of food manufacturing by the processor, and in sourcing of food products by the retailer combine to assure the safety of food to the consumer. This paper describes the various classes of chemicals which can arise as residues in food, particularly agrochemicals and veterinary drugs, the usage patterns which decrease/increase the likelihood of the occurrence of residues at unacceptable levels, how and in what form(s) residues can occur in foods, and the contribution of food processing to a reduction or otherwise of residue levels in food consumed. Consideration will be given, also, to quantifying the risk posed by chemical residues in food and to a study of the results from a Residue Database and from residue monitoring/surveillance as an indicator of risk to the consumer.

Notes: This paper is concerned with processes and procedures involved in the contamination of beef, lamb and pork carcasses during slaughter. The hides of beef and the fleece of sheep are major sources of carcass contamination. The spread of pathogens from beef hides to the carcass, operatives and surfaces in the abattoir is demonstrated. Efforts to clean the hide of cattle and the fleece of sheep are outlined, with reference to the success of these treatments in reducing carcass contamination. The effect of bringing very dirty or dungy animals to slaughter is considered in terms of the effect on carcass contamination after slaughter. The influence of tying the bung (or rectum) in reducing carcass contamination is discussed, as is the use of plastic bags as an additional control in preventing pathogen spread on pig carcasses. The relationship of this revised procedure in reducing the occurrence of yersiniosis in Norway is shown. The use of a commercially automated system to tie beef bungs is discussed in relation to reducing carcass contamination. A comparison between the removal of faecal contamination on carcasses by trimming or using a new steam-vacuumized system is presented. The effect of preevisceration washing of beef carcasses is described and the rationale relating to bacterial removal using this treatment is discussed. The influence of evisceration as a source of carcass contamination is demonstrated in relation to sheep slaughter. The processes of carcass decontamination using washing with water at different temperatures, steam pasteurization and hot lactic acid are compared in relation to their ability to remove bacteria from beef carcass surfaces. Finally, the effect of line speed and the impact of technology advances on beef and sheep carcass contamination is reviewed.

Notes: During the last few years there is a growing feeling in the Dutch livestock and meat industry that it is essential to change from a product oriented to a more market-oriented production (from quantity to quality). This is mainly due to developments such as a more intense international competition, stagnant or declining meat consumption and negative pressure on the consumers’ image of meat. Counteracting these developments is only possible if every part of the production chain contributes (in close cooperation with each other) to achieve a better control of the whole production process: from conception to consumption. For a number of years an extensive research program has been carried out to set up a system of Integrated Quality Control (IKB). This has led to a “total quality concept” which includes the safety and quality of the product, the production method, care for the environment, and human and animal health. This paper discusses the basic elements (structure and content) of this quality concept. The extent of implementation of the IKB-concept depends on the particular sector, but it is at around 70% for pigs and broilers. For IKB in veal, current participation is over 90%.

Notes: The need for residue testing and controls basically derives from two imperatives, the safety of the food supply and requirements imposed in trade. Maximum residue limits (MRLs) for pesticides, veterinary drugs, intentional food additives and potential contaminants, such as mycotoxins, are set for various food commodities based on toxicological assessment. Regulatory testing on domestically produced goods reveals the prevalence of a particular residue in a specific food commodity and also serves as a means to assess the effectiveness of the product registration system, while testing conducted on imports is primarily a verification of the effectiveness of the regulatory program in the exporting country. For industry, testing is required to demonstrate due diligence or to ensure that a product meets the specifications of an export market or a specific contract. Testing programs should be appropriate to meet the intended use. A survey to determine incidence of violations requires a test that detects residues above the MRL, while a dietary exposure study requires methodology that detects residues with the maximum achievable sensitivity. Tests conducted to certify a product for export or to fulfill the requirements of a contract should ensure that the product is in compliance with the market or contract requirement. If the requirement is “no detectable residues”, a very sensitive test method may be required. In many countries, there is a developing consensus that while government has a role in conducting work that is in the public interest, such as ensuring the safety of the food supply within a level of statistical assurance, it is ultimately the producer of the food who is responsible for the safety of the product or for ensuring that the product meets market or contract requirements. Various aspects of this changing regulatory, trade and consumer environment will be considered, with examples of the roles now played by governments and industry to ensure the safety and quality of food products.

Notes: A simulation model that assesses the risk of acquiring salmonellosis from consumption and handling of chicken was created in an Excel™spreadsheet and was simulated using Risk™. The model simulated the distribution, preparation, and consumption of 1,000 chickens and was designed to determine the relationship between the level of Salmonella contamination on chickens at the processing plant exit and the risk of salmonellosis for consumers of the chickens. A scatter plot of the probability of acquiring salmonellosis from consumption of the chickens simulated versus the Salmonella load on the chickens at the processing plant exit clearly showed that highly (i.e., 〉 100 Salmonella/bird) contaminated chickens at the plant exit did not necessarily pose greater risk of salmonellosis than lightly (i.e., 〈 10 Salmonella/bird) contaminated chickens at the plant exit. Rather, greater risk of salmonellosis was realized from lightly contaminated chickens when they were temperature-abused, undercooked, and consumed by someone from the high risk population.

Notes: Transmissible Spongiform Encephalopathies, or Prion diseases, are fatal neurodegenerative diseases that affect both humans and animals. Examples include Creutzfeldt-Jakob disease (CJD) in humans, Bovine Spongiform Encephalopathy (BSE) in cattle and scrapie in sheep and goats. They are characterized by a long incubation period, ranging from months to years, and a variable length clinical course, both determined by the particular infection and the species involved. Some 170,000 cattle in the UK have been diagnosed with BSE and have been destroyed. In the rest of Europe there have also been cases of BSE, though the numbers involved are much lower. The recent potential link between BSE and a new variant form of CJD has highlighted the importance of controlling both BSE and scrapie in Ireland. Control of the disease in both cattle and sheep is complicated by the long incubation period, the lack of suitable tests and the general lack of detailed information in relation to the biology of the disease. Factors that must be considered in designing an effective control strategy include (1) better understanding of the epidemiology of the disease, (2) developing rapid and reliable tests for the disease, (3) introducing a suitable surveillance system for both BSE and scrapie, (4) minimizing the consequences for farmers that have BSE-infected animals, and (5) implementing and monitoring legislation to prohibit the spread of potentially infectious material. Each of these areas is discussed and it is proposed that the introduction of best available technology in both the diagnosis and surveillance of the national herd could form the basis for an effective program against animal prion diseases.

Notes: This paper describes an outbreak of E. coli O157:H7 that occurred in central Scotland. Edipemiological and microbiological evidence has indicated that the outbreak was comprised of several separate but related incidents, relating to the lunch (attended by around 100 people) held in Wishaw Parish Church Hall, a birthday party held in a public house on November 23, 1996 and retail sales in Lanarkshire and Forth Valley. All isolates of E. coli O157:H7 from individuals in the outbreak belonged to phage type 2 and possessed the verocytotoxin gene VT2. Two hundred sixty-two of these isolates have been subjected to pulsed-field gel electrophoresis (PFGE): all had indistinguishable profiles. There were 496 cases in total. There have been 20 deaths (all adults) associated with the outbreak, the highest number of deaths associated with an outbreak of E. coli O157:H7 infection in the world. Of these, 8 people had attended the luncheon served at Wishaw Old Parish Church on November 27, 1996. The age range of those who died was 69 to 93 years. The outbreak was investigated by the Pennington Group who concluded that the circumstances of the central Scotland outbreak, and the inevitable uncertainties that may surround outbreaks generally, provide strong justification for the precautionary, preventative measures proposed and the recommendations made.

Notes: The design and improvement of products and processes often calls for experiments where several response variables are analyzed simultaneously. Frequently, some of these variables are sensory attributes that can only be measured subjectively, through sensory evaluation panels or using expert opinion. In this paper we apply a multiresponse optimization procedure presented in Fogliatto and Albin 1997, to optimize a military food product where 24 sensory attributes are evaluated through descriptive analysis techniques. Our objective is to select the best design and operating control factors considering all attributes simultaneously.

Notes: In a double-blind crossover study, 94 symptomatic adult patients with allergic rhinitis rated sensory preferences for single exposures (2 sprays in each nostril) to three intranasal corticosteroids: triamcinolone acetonide AQ, beclomethasone dipropionate AQ, and fluticasone propionate. Each patient was given the medication and then immediately asked to use a 100-point scale to rate 13 attributes: comfort to take the medicine, medicine run-off, irritation, strength of urge to sneeze, strength of odor, liking odor, strength of taste, strength of bitter taste, liking taste, and moistness of nose and throat, and then (at least 2 min after administration) irritation, medicine run-off, and overall liking. The order of drug presentation was randomized across patients and a 30-min rest interval occurred between drug administrations. Triamcinolone was lower in odor strength than both beclomethasone and fluticasone (mean scores of 8 vs 57 and 52, respectively; P〈0.001) and the odor of triamcinolone was preferred to the odor of the two other sprays (mean score of 72 vs 55 and 55, respectively; P〈0.001). While the taste intensity ratings of the three sprays were not significantly different, the triamcinolone taste was liked more than that of fluticasone (mean scores of 55 vs 41, P〈0.04). Overall, triamcinolone was liked more than fluticasone (P〈0.05). In summary, patient preference testing has shown that triamcinolone is preferred over beclomethasone and/or fluticasone in terms of odor and taste. Such preferences may be important in treatment compliance and outcomes.

Notes: Milk powders available commercially are of different qualities. The quality deviations may be due to inherent quality of the milk which in turn is dependent on various conditions like the fat content, the processing parameters, the storage time and the packaging materials used. The various brands of milk powders have been profiled sensorily for odor and flavor and also positioned using Canonical Discriminant Analysis (CDA). The study throws light on the relative qualities of the different brands, their positive and negative points and their relative position with respect to each other. The results indicate that the various types like whole milk powder, partially skimmed milk powder, skimmed milk powder and infant milk formula cluster together into the same group but the groups are separate from each other. The dominant quality attributes for the groups are different and are discussed in relation to the directional vectors.

Notes: Differences in sensory acuity and hedonic reactions to products lead to latent groups in pooled ratings data. Manufacturing locations and time differences also are sources of rating heterogeneity. Intensity and hedonic ratings are ordered categorical data. Categorical responses follow a multinomial distribution and this distribution can be applied to pooled data over trials if the multinomial probabilities are constant from trial to trial. The common test statistic used for comparing vectors of proportions or frequencies is the Pearson chi-square statistic. When ratings data are obtained from repeated ratings experiments or from a cluster sampling procedure, the covariance matrix for the vector of category proportions can differ dramatically from the one assumed for the multinomial model because of inter-trial. This effect is referred to as overdispersion. The standard multinomial model does not fit overdispersed multinomial data. The practical implication of this is that an inflated Type I error can result in a seriously erroneous conclusion. Another implication is that overdispersion is a measurable quantity that may be of interest because it can be used to signal the presence of latent segments. The Dirichlet-Multinomial (DM) model is introduced in this paper to fit overdispersed intensity and hedonic ratings data. Methods for estimating the parameters of the DM model and the test statistics based on them to test against a specified vector or compare vectors of proportions are given. A novel theoretical contribution of this paper is a method for calculating the power of the tests. This method is useful both in evaluating the tests and determining sample size and the number of trials. A test for goodness of fit of the multinomial model against the DM model is also given. The DM model can be extended further to the Generalized Dirichlet-Multinomial (GDM) model, in which multiple sources of variation are considered. The GDM model and its applications are discussed in this paper. Applications of the DM and GDM models in sensory and consumer research are illustrated using numerical examples.

Notes: A sensory panel utilized chicken patties from Pectoralis major muscle to develop a descriptive vocabulary for warmed-over flavor. Patties were subjected to 4 different cooking temperatures and stored at 4C for up to 4 days. A list of 33 descriptive terms, developed from the literature and a preliminary sample evaluation was modified over 5 daily sessions to 18 terms with corresponding references. In the term selection process the criteria used were, that terms should; have relevance to the product, discriminate clearly between samples, be nonredundant, and have cognitive clarity to the assessors. A comprehensive vocabulary was developed that described the sensory dimensions present in the samples. Over sessions the panel was found to display dynamic changes in their use of the sensory vocabulary. Overall, panel discriminative abilities were enhanced and a process of sensory learning was observed.

Notes: The objective of this study was to determine if hedonically different sweetness levels in yogurt, determined by the ideal-relative rating methods, affected the consumption of yogurt in a sensory specific satiety test. Fifteen subjects attended a preliminary session, six taste tests and three sensory specific satiety tests. During each taste test, subjects rated yogurt with one of the three levels of sweetness (high, optimum, and low) for six attributes. During each sensory specific satiety test, subjects were offered a large serving of yogurt with one of the sweetness levels. Liking for optimum or high sweet yogurts decreased more after consumption than the liking for the low sweet yogurt, however, the consumption of optimum and high sweet yogurts was also greater, thus confounding the experiment.

Notes: The effect of a warm-up sample on reliability of attribute intensity ratings from descriptive panel evaluations was studied by: (1) using a warm-up sample prior to test samples (WU), (2) using a warm-up sample prior to test samples and providing panel consensus attribute ratings for the warm-up sample to panelists (WUC), and (3) providing no warm-up sample (NWU). The standard deviation of ratings from 7 and 4 of 11 attributes for all samples was smallest in WUC and WU, respectively. In paired t-tests, significant differences were found between duplications in 0, 1 and 4 of 11 attributes in WUC, WU and NWU, respectively. Regression analysis of percent fat vs attribute intensity ratings showed the largest R2 from 6, 2, and 2 of 10 significant regression models in WUC, WU, and NWU, respectively. To maximize reliability of descriptive analysis ratings, WUC should be used.

Notes: The beta-binomial model is combined with a Thurstonian psychometric function to obtain estimates of the parameters of a distribution applicable to replicated difference tests. A method of estimating the variance of d’obtained from these tests is provided. A formula for determining sample size, which is composed of the number of trials (or panelists) and the number of replications, to determine d’is also given.

Notes: The effect of flavor types (natural and artificial), aspartame (APM)/acesulfame potassium (Ace-K) blend ratios (50%/50% - 80%/20% sweetness contribution), and acid levels (50-70% dry basis) on flavor profiles of raspberry beverages was evaluated. Optimum flavor profiles of beverages sweetened with APM/Ace-K blends would have high fruit flavor scores, balanced sourness and astringency and minimal side tastes/aftertastes. Descriptive flavor profilists developed a lexicon of flavor/mouthfeel attributes for evaluating the samples in the design. The design was replicated 3 times. Acid level influenced sourness and sweetness scores; increasing acid levels increased sourness and slightly decreased sweetness. Natural flavor in beverages showed lower raspberry intensities across all acid levels and blend ratios than artificial flavor. APM/Ace-K blend ratio had no effect on flavor/mouthfeel attributes. Additionally, HIS side tastes were minimized across all APM/Ace-K blend ratios. The optimum natural flavor beverage would contain a 60/40 to 75/25 APM/Ace-K ratio with 58-70% acid (dry basis). Optimum artificial flavor beverages would contain 50/50 to 80/20 APM/Ace-K ratios with 57-68% acid (dry basis).

Notes: Two simple methods were followed to determine detection thresholds for the taste of substances in aqueous solution. The methods applied were: a modification of the ascending method of limits and a method based on the use of scales. Detection thresholds were calculated for the four basic tastes (sweet, salty, acid, and bitterness), umami and metallic. Reference substances for each taste were sucrose, sodium chloride, citric acid, caffeine, monosodium glutamate and iron (II) sulfate heptahydrate and the results of the two methods were compared. We found that the threshold values calculated by method ASTM-679 was within the range of concentrations identified with the scales method.

Notes: Proteins in saliva samples taken from twelve subjects immediately before and immediately after tasting astringent solutions were analyzed by high performance liquid chromatography. Although considerable variation in both the qualitative and quantitative protein pattern was observed among individuals, three peaks appeared to be common to the majority of the chromatograms. The tasting of the astringent solutions resulted in detectable changes in the protein profile of the saliva samples, observed in all the chromatograms examined, and attributed to the interactions of the saliva proteins with the astringent phenolic compounds. The appearance of a major peak at 8 min after the tasting of the astringent compounds common to most of the saliva chromatograms was attributed to the formation of soluble protein-phenol complexes.

Notes: The perceived textural attributes of an English Cheddar were measured by a trained panel at various intervals during ripening, including those corresponding to mild and medium maturation times for this variety. The moisture content and pH were also monitored. After the recommended ripening period of 50 weeks the matured cheese was significantly less springy, firmer, harder, crumblier and creamier, indicating that textural attributes are related to the age of the Cheddar. Measures beyond the recommended maturation period revealed further significant differences. Comparisons of the measures taken at mild, medium and full maturity also revealed differences in the textural properties of the different classes of Cheddar. Significant correlations (p ≥0.01) between pH and springiness, crumbliness by fingers and creaminess suggested a strong relationship between these textural attributes and the extent of proteolysis. The results of this study revealed a strong relationship between age and the textural attributes of cheddar when restricted to one particular variety.

Notes: Sensory research on kiwifruit has shown that specific sugars and acids, and volatile flavor compounds have causative effects on “sweet” and “acid” aroma and flavor attributes. New Zealand consumers (n = 162) assessed the perceived flavor intensity and acceptability of three volatile flavor compounds (ethyl butanoate, E-2-hexenal and hexanal) at varying levels in a model base solution. Increasing levels of ethyl butanoate positively affected all the acceptability attributes (“overall liking,”“liking of aroma,”“liking of flavor”) as well as increased the perceived intensity of “kiwifruit aroma” and “kiwifruit flavor.” Increasing levels of E-2-hexenal negatively affected all the acceptability attributes but increased the perceived intensity of “kiwifruit aroma” and “acid flavor.” An increase in hexanal increased perceived intensity of “kiwifruit aroma.” Ethyl butanoate and E-2-hexenel at the levels tested had the most prominent effects on consumer perceived intensity and acceptability of the “kiwifruit flavor.”

Notes: Wine vinegar is a product obtained from wine acidification which contains at least 5% by wt. of acetic acid, in general without any additives or colorings.Aspects studied in this work include: the determination of the taste group thresholds (geometric mean of the individual best-estimate thresholds “BET”) of two different acids (citric and acetic acids) in aqueous solution and spanish vinegars produced from table and sherry wines. The results obtained suggest that wine vinegar can be considered something more than just an acidulant agent.In order to evaluate differences among wine vinegars, discriminant tests for twenty-five spanish vinegars (sherry, table and flavored vinegars) were applied. Six of the twelve attributes freely chosen by assessors allowed grouping of the spanish wine vinegars according to their sensory aspects.

Notes: Expectations are generated by a variety of factors. We indicate a flow chart for the role of expectations at the point of choice and in influencing sensory perception at the time of consumption. We review the sparse literature on how advertising, packaging and information generate sensory expectations. The application of various theories to explain the observed effects of sensory expectations are reviewed. There is overwhelming evidence for assimilation-contrast effect, although no studies have been specifically designed to detect it. Finally we review the reasons why individuals might differ in the way that expectations influence sensory perception. These reasons include ideas from persuasion literature and private body consciousness. A number of behavioral hypothesis that follow from these theories are developed.

Notes: Seventy-nine children ages 36–71 months evaluated 2% commercial UHT milk and 2% commercial pasteurized milk for taste, color and mouthfeel with age appropriate facial hedonic scales with Peryam and Kroll verbal descriptors. Children significantly preferred the pasteurized milk over UHT milk for taste and mouthfeel attributes. Descriptive analysis of milks with an adult trained panel showed a significantly higher cooked note in UHT milk products which may explain the difference in preference. The results also showed that children 36–47 months were able to use the 3-point scale; children 47–59 months the 5-point scale; and children 60–71 months the 7-point scale to score preferences for food. Data transformation for a 0 to 100 point scale provided ease of comparison between age groups. Using the appropriate testing instruments, young children were able to provide valid food preference information.

Notes: This paper compares experts and consumers to determine the degree to which they agree with each other on ratings of 37 sauce products, using the same sensory attributes. The paper also assesses the degree to which sensory attribute ratings correlate with objective physical measures. The ratings of experts (1-9 scale) and consumers (0–100 scale) agree quite highly, as shown by the high correlation between the two panels across the 37 products. The paper refutes the notion that consumers are incapable of validly rating the sensory aspects of products. The paper therefore presents the case for using consumers to assess the sensory characteristics of products.

Notes: The purpose of this study was to assess the sensory characteristics of sodium lactate (SL) in combination with sodium tripolyphosphate (STP) in a model system at common usage levels for meat products. Using a gelatin-based (2%) system containing various concentrations of sodium lactate (0%, 1%, 2%, or 3%) and sodium tripolyphosphate (0%, 0.1%, 0.2%, 0.3%, or 0.4%) a thirteen-member trained sensory panel scored samples for saltiness, bitterness, and soapiness. As sodium lactate and sodium tripolyphosphate levels increased, saltiness increased. Bitterness decreased with addition of 1% SL and increased with 2 and 3% SL. Soapiness also increased with increasing levels of SL. Bitterness and soapiness were not affected by increasing STP concentrations.

Notes: The impact of 2-methylisoborneol (MIB) (earthy/musty), on the flavor evaluation of farm-raised catfish was assessed by a descriptive analysis panel, who are trained to evaluate flavor intensity using universal scales. Ideally, panelists should give the same intensity ratings for all attributes within a sample. Realistically, individual panelists perceive flavor intensity differently, because of variations in detection thresholds, adaptation, fatigue and enhancement or suppression. Experiments were designed to investigate problems associated with evaluating catfish flavor. Samples “spiked” with MIB had lower chickeny flavor than unspiked. Intensities of MIB in spiked samples that followed an equally spiked sample were lower. Problems can occur when catfish are presented at intervals less than 7 min. Panelists experienced difficulty in determining intensity differences of MIB flavor between sessions, but could determine differences in intensity of MIB within a session. These influences must be accounted for in experimental designs to assess catfish flavors.

Notes: Salivation may affect texture and taste perception. Thus, any influences that affect salivation also might affect a panelist's sensory responses. Twelve sensory panelists were compared for psychophysiological factors that might influence their salivary responses. Medication, body mass index (Quetelet), dieting-induced deprivation, nutrient deficiency, and caffeine, tannin, and fiber consumptions along with smoking were considered. Specific nutrient deficiencies or fiber consumption did not affect salivation of the panelists studied. All other factors, especially smoking and diet-induced deprivation, appeared to warrant further study with a larger population.

Notes: The objective of this study was to compare data obtained using descriptive analysis (DA) and free-choice profiling (FCP) of a simple model solution. Three menthol isomers (l-, d-, dl-), each at 0.01, 0.02, and 0.04% (w/v), were tested. A trained DA panel (n = 11) used two aroma, three nasal-sensation, two taste, and five mouth-sensation terms to describe the samples. An untrained panel (n = 15) used FCP to assess the same solutions. Individual FCP panelists used from nine to twenty-one terms. All DA terms significantly discriminated among the samples. Seven to eighteen of the FCP panelist descriptors significantly discriminated among the samples. Procrustes analyses (PA) indicated that FCP and DA similarly discriminated among the samples, possibly because the solutions were essentially one dimensional. Sample loadings on dimension 1 of both spaces were correlated (r = 0.966) as were dimension 2 loadings (r = 0.783). The FCP Rc-value was 91.2% and the DA Rc value was 85.6%. At higher concentrations (0.04 and 0.02%) l-menthol was significantly stronger than d-menthol in eucalyptus aroma, nasal clearing, all mouth cooling attributes, and bitter attertaste. At 0.01 % concentration no significant differences were detected among the menthol isomers.

Notes: Nonparametric models for sensory discrimination methods are developed. In these models, the relationship between the probability of a correct response, Pc, and the measures for sensory discriminability or sensory difference, p1 and p2 are given. On the basis of the models, different sensitivities for triadic tests were compared including 3-AFC and triangle methods with the strong stimulus as the odd sample and with the weak stimulus as the odd sample. Predictions were made which can be tested experimentally.

Notes: A table is provided for testing the significance of R-index values. The use of the table is illustrated and the theory and assumptions behind the method are explained.

Notes: There are many ways in which a laboratory difference test differs from ‘real life’ discrimination of foods. One of these is the interval of time between tasting the two stimuli to be discriminated. To investigate this, judges performed same-different discrimination tests using a citrus flavored beverage as a medium. The time interval between tasting the standard and comparison stimuli was varied. In this initial study, short intervals of zero, 30, 60 s were examined. For judges unfamiltar with the stimuli, performance deteriorated as the time interval increased. For judges familiar with the stimuli, the zero time interval elicited best performance but there was no decrease in performance when the interval was increased from 30 to 60 s. The results were explained by hypothesizing different types of memory trace for the standard stimulus being utilized for comparison with the comparison stimulus.

Notes: To determine quality characteristics of snack chips, six snack chip prototypes containing cowpea flour, cornmeal, wheat flour and cornstarch (26, 34, and 42%) were formulated. Desirable characteristics as determined by three focus groups were a golden yellow color in a thin, smooth corn-chip form with crunchy or crispy texture, greaseless or dry mouthfeel and salty corn flavor. Chips were undesirable if brown-colored, thick, oily or greasy, had either hard or crumbly texture, and had bland, fishy, or burnt flavors. In consumer acceptance tests, commercial products were found to be more appealing than the experimental products. A discriminant model based on instrumental measurements of color and texture from 11 snack chip samples (six prototypes and five commercial samples) correctly grouped 100% of the acceptable and 90% of unacceptable products.

Notes: To find selective and predictive tests in texture profiling, a series of 20 tests and 2 texture profiles were submitted to 25 subjects. Selection was carried out according to ranking, scoring and a texture property knowledge questionnaire. Two profiles were then done on 5 Bolognese sauce and 8 rice samples. Assessors were assessed by Spearman correlation coefficients for ranking tests, F values for scoring, average square canonical correlation coefficients for measuring the dimensionality of individual sample space, product discrimination and response repeatability for both Bolognese sauce and rice samples.Characterization test results showed that subject ability varied widely according to the test and the profile. Only one significant relationship between the texture property knowledge test and rice profile performances was found. More tests were successfully related to one or several texture attributes of both profiles. The ability to complete a complex profile could be discerned, albeit with difficulty, through simple selection tests.

Notes: Instrumental measurement of color places hue in a 360° continuum, but sensory description of color usually refers to distinct hues — red, green, yellow etc. Color evaluation of “Golden Delicious” apples by both an instrumental method and an experienced sensory panel was conducted to determine if sensory panelists perceive changes of golden apple color in green and yellow hues separately or on a hue continuum from green to yellow. Apple color changed from green to yellow during storage. Two different sensory scoring ballots were used to determine if sensory panelists view color in discrete green and yellow hues or on a green-to-yellow continuum. Data collected from this study demonstrated that sensory panelists can detect the color changes of decreasing greenness, increasing yellowness, and color change from green to yellow in an equal fashion. Hue angle represented the best predictor of sensory color perception.

Notes: Interindividual differences in sweet and bitter taste sensitivity were investigated using time-intensity (TI) measurements and multivariate statistics. TI profiles were obtained in triplicate from 25 subjects for 23 sweet and/or bitter stimuli first matched to be approximately equi-intense to 200 mM NaCl. Sweet stimuli, except for the larger sweeteners, were less persistent, and required less time to reach maximum intensity than bitter stimuli. The results of principal component (PCA) and cluster (CA) analyses of the stimuli X subjects matrices for maximum intensity (Imax), time to maximum intensity (Tmax), total duration (Tdur), and area under the curve (Area) suggest that sweet and bitter stimuli do not share common receptors; and that there are at least two receptor mechanisms each for sweet taste (one for sugars and other small compounds, and the other for large sweeteners) and bitter taste (one for PTC/PROP and one for other bitter compounds).

Notes: The objective of this study was to assess the sensory characteristics of potassium lactate (PL) in combination with salt (NaCl) in a model system at common usage levels for meat systems. Using a gelatin-based (3%) model system containing various concentrations of potassium lactate (0%, 1%, 2%, and 3%) and sodium chloride (0%, 0.5%, 1%, 1.5%, 2%, 2.5%, and 3%), a sensory panel (10) selected for salt sensitivity (0.08% threshold or less) scored the intensity of saltiness and bitterness of each treatment combination using a scale of 0 = none and 15 = intense. As salt concentration increased, saltiness increased and bitterness decreased; as PL increased, saltiness and bitterness increased. At 2% salt, the bitterness of PL was masked; however, salt concentrations below and above 2% were less effective in bitterness reduction.

Notes: Multidimensional scaling (MDS) procedures produce maps of stimulus points, the dimensions of which may be sensory and/or physicochemical attributes. Our objective was to test the efficiencies of a cyclic design and a MDS method in the study of saltiness perception. Two levels of NaCl (0.1% and 0.2%, w/v) were added to two nonionic and two ionic gum solutions prepared to two concentrations. Subjects scaled dissimilarities between pairs of solutions and rated each sample for flavor attributes. Solution pairs were selected using cyclic designs. Repeated measures ANOVA determined that added NaCl was the only significant factor affecting saltiness perception. In contrast, from KYST-2A MDS maps, we concluded that saltiness perception was influenced by gum property, gum concentration, presence of calcium and potassium, and related to binding of the sodium ion (Na+) as determined by23Na nuclear magnetic resonance (NMR) spectroscopy. The MDS cyclic design successfully reduced the number of samples and subjects when using experienced subjects.

Notes: Parameters of freezing of sucrose solutions and emulsions were evaluated using differential scanning calorimetry (DSC) at cooling rates of 1 to 20°C/min. The differential and cumulative nucleation spectra of extracellular ice nucleators (ECINs) were established using a w/o emulsion technique. Freezing curves were obtained in a cabinet at −7 and −12°C. The ECINs increased the ice nucleation temperature and reduced the time of the supercooling stage. Although the time of phase transition increased in the presence of ECINs, the total time of freezing decreased. The ECINs caused lower onset temperatures of freezing as compared to the original cells.

Notes: Performance of thermal process simulation software was tested for predicting internal product temperature and lethality. Responses were tested for process deviations on canned products with a wide range of heating characteristics for computer-based on-line control of retorts. Static and agitated processes were tested with products exhibiting heating rate factors (fh) ranging from 2 to 70 min. Cans were fitted with thermocouples and subjected to deviations of various types. Center temperature profiles and lethalities predicted by the model in response to dynamic retort temperatures were compared with those measured by thermocouples. Profiles agreed, and process lethalities calculated from predicted and measured profiles agreed ±10% with the model slightly underpredicting measured lethality.

Notes: Our objective was to determine if chlorophyll fluorescence could be used as an indicator of physiological changes in cold-stored broccoli after transfer to room temperature. Mass loss, respiration, and ammonia content increased when the broccoli were held at 18°C for 72h, whereas the chlorophyll fluorescence measurements (Fv/Fm, T½ and Fmd) decreased. Broccoli color began to change after 24h, as hue decreased rapidly and chroma increased. Fv/Fm decreased markedly only after the first 24h when the color changes began, whereas T½ and Fmd decreased continuously as the broccoli aged. Both T½ and Fmd correlated negatively with mass loss and respiration.

Notes: Air-classified pea protein was texturized using a twin-screw extruder. The effects of moisture, screw speed and barrel temperature on the physical, functional and nutritional characteristics of texturized pea protein were investigated. Increased dough moisture increased product bulk density (BD) and available lysine (LYS) of texturized pea products, but decreased water-holding capacity (WHC) and in vitro protein digestibility (DIG). Raising the screw speed reduced BD and LYS of texturized pea protein, but increased WHC and DIG. Bulk density and LYS decreased as barrel temperature was increased, whereas WHC and DIG increased. Texturized pea protein resembled commercial texturized soy products with respect to most physical and functional properties.

Notes: Pre-cooked roast beef slices were stored 28 days at 4±2°C in air or 100% N2 with and without vaporized horseradish essential oil (HEO). Addition of 20 μL HEO/L restricted growth of most spoilage bacteria. Pseudomonas spp. and Enterobacte-riaceae were strongly inhibited by HEO. Lactic acid bacteria were more resistant to the antimicrobial effect and dominated spoilage flora. Sensory evaluation and headspace analysis by gas chromatography/mass spectrometry revealed that development of off-flavors and odors derived from fat oxidation products was delayed by HEO. Cooked meat color was also preserved in samples stored under HEO.

Notes: The effect of electrolyzed water on total microbial count was evaluated on several fresh-cut vegetables. When fresh-cut carrots, bell peppers, spinach, Japanese radish, and potatoes were treated with electrolyzed water (pH 6.8, 20 ppm available chlorine) by dipping, rinsing, or dipping/blowing, microbes on all cuts were reduced by 0.6 to 2.6 logs CFU/g. Rinsing or dipping/blowing were more effective than dipping. Electrolyzed water containing 50 ppm available chlorine had a stronger bactericidal effect than that containing 15 or 30 ppm chlorine for fresh-cut carrots, spinach, or cucumber. Electrolyzed water did not affect tissue pH, surface color, or general appearance of fresh-cut vegetables.

Notes: Storage conditions of part-baked brown soda bread were studied as related to changes of crust and crumb moisture during rebaking at in-oven temperatures of 180 and 200°C for 10–40 min. Baking loss during rebaking originated solely from the crust area, whereas the moisture content of the crumb remained constant. Packaging in an atmosphere of 40% CO2 and 60% N2 and storage at 4°C inhibited microbial contamination for 13 wk. The relationship of moisture content and water activity of part-baked breads followed the characteristic of a sorption isotherm and was mathematically described by either a quadratic or exponential function.

Notes: Heat-processed and fresh salsa formulations were developed with different levels of honey (up to 10%), acid, and capsaicin. Descriptive sensory characteristics and consumer acceptability of the salsas were studied. Increasing levels of honey increased sweetness and crispness of both salsas. Higher levels of honey decreased oral heat intensity more than did lower levels. Consumers liked heat-processed salsa better than fresh and acceptability generally decreased with increasing honey content. However, about one-third of consumers liked salsas with honey more than samples without honey.

Notes: Solubility, electrophoresis, viscosity and emulsifying properties of heated solutions of yolk, plasma and granules were determined. Plasma and yolk were not affected when heated under 69°C. Above 69°C, protein solubility dropped sharply and apparent viscosity rose sharply because of aggregation of proteins. For granules, protein solubility and apparent viscosity were not modified up to 76°C. The constituents of granules were not denatured. Emulsifying activity of yolk and plasma decreased after heating at 72°C but remained steady for granules. Emulsion stabilization properties of yolk, plasma and granules were not influenced up to 76°C. Results suggested that intact granules withstood more severe heat treatments than egg yolk without lessening their emulsifying properties.

Notes: Each of three cheese batches was randomly divided into two parts, one was subjected to slow freezing for 15 days and the other maintained under refrigeration. Thereafter the cheeses were tempered under refrigeration for 1 mo and evaluated weekly. In general, cheeses presented low melting capacity as a result of low levels of proteolysis. This low intensity of proteolysis was probably a function of the destruction of the lactic starter and partial inactivation of the coagulant during stretching of the cheese. Free oil content increased during storage for both treatments, but more in the tempered samples.

Notes: Bovine meat juice was used as a model system to examine the kinetics of the formation of heterocyclic amines. The formation was examined at temperatures between 100 and 225°C for 0.5-180 min. Heated samples were subjected to solid-phase extraction and HPLC analysis. IQx, MeIQx, 7,8-DiMeIQx, 4,8-DiMeIQx, PhIP, harman and norharman were identified. The formation of the IQx derivatives and PhIP followed a first-order model Ct=B(1–ek1(t1-t0), and the temperature dependence followed the Eyring equation k=kbT/heΔS/Re-ΔH/RT. The parameters in the first-order model and the Eyring equation could be estimated from our data. The degradation of heterocyclic amines at high temperatures was analyzed and the products were found to be less mutagenic.

Notes: Irradiation dose affected production of volatiles in vacuum-and aerobic-packaged cooked pork sausage, but its effect on TBARS was minor. Storage increased production of volatiles and changed their composition only in aerobic-packaged sausage. Among volatile components, 1-heptene and 1-nonene were influenced most by irradiation dose, and aldehydes by packaging type. TBARS and volatiles of vacuum-packaged irradiated cooked sausage did not correlate well. However, TBARS had very high correlation with amount of aldehydes, total volatiles, ketones and alcohols with long retention times in aerobic-packaged pork sausage. Heptene and 1-nonene could be indicators for irradiation; and propanal, pentanal, and hexanal for oxygen-dependent changes of cooked meat.

Notes: Highly visible white and greyish deposits appeared on commercially pickled green asparagus spears after 6 mo storage. Microscopic examination revealed that the deposits consisted mainly of needle shaped crystals. Rutin (querce-tin-3-0-rhamnoglucoside), present in asparagus in appreciable quantities (0.02 to 0.1%) was suspected of being responsible for the deposits. The deposits were chromatographed with authentic rutin using HPLC. Retention and spectral data confirmed that the major component of the deposits was rutin.

Notes: Pecan texture was measured by texture profile analysis (TPA) at 80% compression before and after being frozen at 6 controlled rates, to 3 minimum temperatures and thawed at 2 rates. Freezing and thawing rates had more influence than storage temperature on pecan texture. Certain TPA parameters (i.e., hardness, area 1, fracturability and slope of linear segment of force vs. deformation curve) were more affected than others by freezing and thawing rates. Freezing and thawing at high rates had the least effect on pecan texture.

Notes: American ginseng roots were treated with an antimicrobial agent, and stored under various CA (2,5, and 8% CO2) or MA conditions to extend the shelf-life. Changes in respiration rates during CA storage, gas composition in packages, saponin and free sugar content, and other quality factors were monitored during storage. The respiration rate of ginseng increased rapidly during the first month of high CO2 CA storage and then slowly decreased to stable levels after 3 mo storage. In MA studies, the equilibrium CO2 concentration was attained after 20 days. There were no noticeable changes in appearance, saponin content and free sugars after 3 mo CA (5% CO2) storage and MA packages.

Notes: The rates were monitored on biomass increase, polysaccharide production and viscosity development of whey broth and a control synthetic broth during fermentation by Rahnella aquatilis and organic acids, lactose, peptides and free amino acids were measured. Growth curves were similar and characterized by maximum specific growth rates of 0.61 h−1 for whey and 0.63 h−1 for synthetic medium. The yields of polysaccharide were 0.59 g/glactose for the synthetic medium and 0.56 for whey. Small peptides (〈4,000 Da) and most free amino acids in both fermentation media were consumed within 24h.

Notes: Growth of Escherichia coli and Staphylococcus aureuson the surface of Trypticase Soy Agar (TSA) packaged with various CO2 partial pressures (0, 20, 40, 60, 80, 100%, balance N2) was compared to the control (N2 100%) on TSA in which the pH was adjusted to equal that in CO2 atmospheres at 15°C and 30°C. At 15°C, the biostatic effect was noted with all CO2 partial pressures for both species. At 30°C, the biostatic effect of CO2 was almost completely nullified for E. coli, but that for S. aureus was still effective. S. aureus was more sensitive to the inhibitory effects of CO2 than E. coli at both the temperatures.

Notes: Nondestructive analysis of water distribution and structural changes in cooked rice grains, Oryza sativa cv Koshihikari, was performed with Nuclear Magnetic Resonance (NMR) Micro imaging of protons (1H). Using a specially designed holder, high quality images of cooked rice grains were obtained, even when up to 10 grains were studied simultaneously. Internal hollows were observed in all examined cooked rice grains, and we propose a mechanism to explain their formation. The origin of these hollows was hypothesized to be cracks or fissures, and hollows resulted from sealing of such lacerations by gelatinized starch in the peripheral layer in combination with expansion of the grain during cooking.

Notes: Experiments were conducted to measure the temperature and moisture content of starch and starch-fructose samples during drying. A two-dimensional finite element model was developed. An exhaustive parameter estimation technique was applied in conjunction with the model and drying data to estimate the moisture filtration coefficient kp and the surface heat and mass transfer coefficients, hq and hm. Minimization of the deviation between experimental and simulation data resulted in a kp value of 0.51*10−7 kg.m/N.h and 0.52*10−7 kg.m/N.h for the starch and starch-fructose samples. Corresponding values of hq were 15.8 W/m2K and 21.2 W/m2K and that of hm were 1.12*10−6 m/s and 0.94*10−6 m/s respectively.

Notes: Fresh camel meat samples were treated by dipping in solutions of sodium acetate (10% w/w), potassium sorbate (1.5% w/w), sodium lactate (5% v/v of 60% solution), or trisodium citrate (1.5%w/w) alone or combined with Bifidobacterium breve cell suspension (5%) and stored at 4°C. The pH, psychrotrophs, and sensory attributes were evaluated. Sodium acetate alone or combined with bifidobacteria maintained pH level, extended microbial shelf life (〉12 days) and minimized surface discoloration. Bifidobacteria cell suspension when applied alone had negligible effects on microbial and sensory attributes. However, when combined with sodium acetate or potassium sorbate, bifidobacteria exhibited an additive effect on suppressing spoilage microorganisms.

Notes: Time domain proton NMR was used to study translational diffusion and molecular mobility of sugar and water molecules in simple aqueous solutions to determine how the addition of hydrocolloid stabilizers affect their mobility. Results showed that addition of Locust Bean Gum (LBG) did not affect the diffusion or mobility of either the sugar or water molecules over distances up to 10μm in unfrozen solutions. The diffusion properties of the sugar and water molecules were not affected by either biopolymer cryogelation or the presence of ice. We hypothesized that retardation of the ice crystal growth rates in aqueous sugar solutions caused by the presence of LBG was not a bulk property of the solution. The most likely cause is a modification of the local interfacial region between the ice crystal surface and the surrounding unfrozen solution.

Notes: We determined the bactericidal activity of surface applied waxes on oranges. Effective bactericidal activity of combined alkali and heat treatments was observed on both glass slides and orange fruit surfaces. A five log reduction of Escherichia coli was attained by dipping inoculated glass slides in heated (≥50°C) alkaline (pH 10) wax solution for 4 min. A pH 11, dipping at ≥50°C for ≥2 min achieved similar bactericidal effects. On the fruit surface, wax treatments were effective at the non-stem scar area. Thus, applied fruit waxes may be useful on raw agricultural commodities to reduce surface microbial contaminants.

Notes: Pork trim-finely textured (PTFT), generated by advanced meat recovery which mechanically removes meat from bones under pressure, has properties that may cause shelf-life instability. Lipid oxidation, microbial growth and pH of retail displayed fresh ground pork patties were not affected (P〉0.05) by addition of up to 15% PTFT. Ground pork with 15% PTFT and 3.3% sodium lactate (NaL) stored in chubs at–2.2°C for 14 days was ground and modified atmosphere packaged (MAP) or overwrapped (OW) and retail displayed for 4 days. Lipid oxidation increased (P〈0.05) due to PTFT incorporation but was reduced by NaL. MAP and/or NaL reduced microbial counts and lipid oxidation. MAP maintained redness (a*) and prevented metmyoglobin formation.

Notes: Fundamental dynamic rheological tests were used to measure the rigidity (|G*|) and viscoelasticity (tan δ) of gels made from skim milk samples that were (1) never cooled, (2) stored at 4°C, or 3) stored at 4°C and then rewarmed to 25°C. Milk cooled and held at 4°C prior to gel formation produced gels with lower |G*| and higher tan(δ) than fresh milk gels. Chilled milk samples that were rewarmed to 25°C for 16h prior to gel formation produced gels with rheological properties similar to fresh milk gels. The solubilization of β-casein during cold storage was hypothesized to be the cause of observed differences in gel rheology.

Notes: We compared processing yield, composition, and quality of tofu from soybeans and from full-fat soyflakes. Tofu was made using a steam-jacketed kettle and a commercial steam-injected cooker. Hydration time was 10 min for flakes and 12h for whole beans. Regardless of cultivar, a higher tofu yield was obtained from the steam-injected cooker system than from the steam-jacketed kettle system. Utilization of flakes required 62--65% less water during soymilk production. Independent of cultivar, tofu produced from full-fat soyflakes was lower in fat (26% d.b.) than whole soybean tofu (40% d.b.).

Notes: Heat stabilities of crude and partially purified soluble (SPOX), ionically bound (IPOX) and total peroxidase (TPOX) from fresh pinto beans were investigated at 55–90°C. Heat inactivation of peroxidase (POX) followed first-order reaction kinetics. Each inactivation curve consisted of two linear parts: initial rapid inactivation (heat-labile) followed by slower inactivation (heat-stable). IPOX showed activation during heat treatment with a highly heat-stable isoenzyme (D90=40 min) which was more heat-stable than SPOX. Activation energies for heat-stable parts of crude IPOX and SPOX were, respectively, 12.1 and 36.4 kcalmol-1 with z values 45.4 and 14.1C°. Heat stable SPOX isoenzymes (D70=22.6) were obtained by 65–95% (NH4)2SO4 precipitation from crude SPOX. Two POX fractions (F1 and F2) were separated from TPOX by ion-exchange chromatography.

Notes: Changes in color of irradiated meat were observed to be species-dependent. Irradiated pork and turkey became redder due to irradiation but irradiated beef a* values decreased and yellowness increased with dose and storage time. The extent of color change was irradiation dose-dependent and was not related to myoglobin concentration. Visual evaluation indicated pork and turkey increased in red ness whereas beef decreased in redness as dose levels increased. Reflectance spectra showed that irradiation induced an oxymyoglobin-like pigment in pork and that both oxymyoglobin and metmyoglobin developed in beef as a result of irradiation.

Notes: Cheddar cheeses with 33% reduced-fat content were prepared with granular soy lecithin, hydrogenated soy lecithin, or oat. Addition of lecithin increased the wet weight yields and moisture of cheeses (P≤0.05). Reflected color values (L* and hab) were increased in cheeses containing granular soy lecithin (P≤0.05). Acid values of cheeses with lecithin were higher than control cheeses (P≤0.05). Concentrations of lecithin at 0.2% (w/w) resulted in visible changes in micro-structure of the cheeses. Granular soy lecithin or oat lecithin added to reduced-fat cheeses resulted in a decrease in flavor quality (P≤0.05). Hydrogenated soy lecithin added to reduced-fat cheeses improved texture quality without negatively affecting flavor quality.

Notes: ‘Chandler’, ‘Oso Grande’ and ‘Sweet Charlie’ strawberries were stored for 8 days at 1 or 10°C, or 4 days at 20°C, either unwrapped or wrapped in PVC film to retard were conducted during the 1 water loss. Total ascorbic acid (AA) content was expressed on a dry weight basis to correct for water loss differences between treatments. Loss of AA was low and did not differ between wrapped treatments at 1 and 10°C, but was much greater at 20°C. Wrapping reduced AA loss by 5-fold at 1 and 10°C and by 2-fold at 20°C. The effect was not due to modification of O2 and CO2 levels in wrapped treatments, which was minimal. The results indicate that water loss had a greater effect on AA levels than temperature. Combining wrapping with storage at 1 or 10°C reduced AA loss by 7.5-fold compared to unwrapped strawberries stored at 20°C.

Notes: Heat transfer characteristics were studied on clarified depectinized peach juices heated in an agitated jacketed vessel. Four types of agitators and several agitation rates were analyzed. Two equations were derived for the density and rheological behavior of peach juice based on the combined effects of temperature and soluble solids content. Overall and internal heat transfer coefficients were also determined. For each agitator, Nusselt number was statistically correlated as a function of the Reynolds and Prandtl numbers, and as the ratio of the fluid viscosity at the bulk temperature to the fluid viscosity at the wall temperature. The power number also correlated with the Reynolds, Froude, and time numbers.

Notes: The effect of moisture content (MC) on texture and properties of extrudates with varied ratios of soybean flour (SF) and wheat flour (WF) was studied. A single-screw extruder was used at screw speed 200 rpm. MCs of blends were 16, 17 or 18%, w.b. The properties of extrudates depended on flow rate of the material during extrusion. The flow rate revealed a nonlinear dependence on the blend composition and the MC at the same volume of filling of the screw feeding section. The expansion ratio of WF or SF extrudates increased with lowering of the MC. Unexpectedly, the expansion ratio decreased with lower MC for the composite extrudates. Optimal extrudate properties at 16% (w.b.) MC corresponded to 80, 90 or 100% (d.b.) WF and 20, 10% or no SF, respectively. At 17 and 18% MC, optimal properties were found for products containing 80 or 90% WF and 20 or 10% SF, respectively.

Notes: Cells of Lactobacillus delbrueckii subsp. lactis I produced hydrogen peroxide at 5°C in sodium phosphate buffer (0.2M, pH 6.5) with or without glucose. However, if the cells were starved by preincubation in buffer alone, glucose or sodium lactate were necessary to cause hydrogen peroxide production at 5°C. Hydrogen peroxide production by nonstarved cells was confirmed to be in part due to a NADH oxidase. The production of hydrogen peroxide by starved cells in buffer plus glucose in the early stage of incubation was associated with the production of a small portion of lactic acid which disappeared upon further incubation. Additional experiments revealed that hydrogen peroxide was produced in buffer containing sodium lactate added in place of glucose. Results suggested the presence of a lactate oxidase in the organism which used D-lactate to produce hydrogen peroxide.

Notes: Soymilk made from lipoxygenase-free soybeans had less cooked beany aroma, less cooked beany flavor and less astringency and was rated darker and more yellow than that made from soybeans with normal lipoxygenase. Sensory descriptive panelists noted no differences between lipoxygenase-free and normal soybeans for milky flavor, wheat flavor, thickness, chalkiness or aftertaste. Tofu made from lipoxygenase-free soybeans had less cooked beany flavor than that made from normal soybeans. There were no differences in cooked beany aroma, raw beany aroma, raw beany flavor, wheat flavor, astringency, hardness, darkness or yellowness. Native-born Japanese, Chinese and U.S. descriptive panelists differed in responses to flavor, texture and color of soymilk and tofu.

Notes: Increasing the concentration of salt (NaCl) or starch in a salt-starch solution depressed the dielectric constant and loss factor relative to a pure salt solution, except for the loss factor when no salt was present. The addition of salt to a starch solution increased the viscosity. Both viscosity and dielectric properties were decreased by mechanical agitation and by action of amylase. Above 10% starch the decrease in dielectric loss was abrupt, corresponding to the increase in viscosity. The changes were hypothesized to be due to an interaction between salt and starch, either through direct binding of the salt, or indirectly through the increase in viscosity.

Notes: Progressive freeze-concentration was effectively applied to concentrate glucose in a model solution. Effective partition constants of the solute between the liquid and solid phases were determined experimentally. The constant was strongly dependent on the stirring rate at the ice front and the advance rate of the ice front. Increasing stirring rate and reduced rate of advance lowered the effective partition constant: an improvement in freeze concentration performance. A concentration polarization model was useful to describe the effects of the operating conditions on the effective partition constant.