Bacteriological Survey of the Frozen Prepared Foods Industry: II. Frozen Breaded Raw Shrimp

During inspection of 21 frozen breaded raw shrimp processors, 861 finished product units (retail packages) and 778 line samples were collected and analyzed bacteriologically. The bacteriological results for the finished product samples collected in plants with poor and good sanitary controls are presented and compared. The samples consisted of 4 to 38 units (retail packages) of the finished product as offered for sale by the processors. Frozen raw breaded shrimp collected from plants operating under good conditions of sanitation had the following bacterial contents: 85% of the samples had an average (geometric) aerobic plate count of less than 1,000,000 per gram; all of the samples had an average (geometric) most probable number of less than 1,000 coliforms per g; 81% of the samples contained less than 20% Escherichia coli-positive units; 57% of the total number of units in 0.1-g portions were negative for coagulase-positive staphylococci, and 95% of the units had less than 1,000 coagulase-positive staphylococci per gram.     

Bacteriological Survey of the Frozen Prepared Foods Industry: III. Potato Products

During sanitation-inspections of 29 potato-processing firms, 2,544 finished product units and 1,654 samples were collected and analyzed bacteriologically. The results of the bacteriological examination of finished French fries, fried potato cylinders, and dehydrated potatoes usually did not reveal the conditions of cleanliness under which they were produced. This was most likely due to the lethal effect of the terminal fry to which the French fries and potato cylinders are subjected, and because of the lethal effect of the dehydration temperatures used in processing the dehydrated potato products. However, as in most food-processing firms, line samples collected at each processing step reflect sanitary conditions and provide bacteriological support of inspectional evidence of plant insanitation. Most of the frozen stuffed baked potatoes examined were produced under poor sanitary conditions. But because this product is usually processed while hot, bacteriological examination of the finished product did not usually reveal the conditions of cleanliness under which they were produced. Again, inspectional observations were necessary for a full evaluation of the conditions of production. In contrast, frozen potato patties and frozen hash brown potatoes were more likely to reveal the conditions of sanitation under which they were produced as evidenced by the varying bacterial counts.     

Bacteriological Survey of the Frozen Prepared Foods Industry: I. Frozen Cream-Type Pies

During Food and Drug Administration inspections of 12 imitation cream pie producers, 453 finished product samples and 350 line samples were collected and analyzed bacteriologically. Sanitary conditions in the plants varied from good to poor and, in general, were reflected in the bacteriological results. The survey revealed that, in the great majority of cases, frozen imitation-cream pies produced in plants operating under good conditions of sanitation had the following bacteriological content: (i) a most probable number (MPN) of fewer than three Escherichia coli cells per gram (i.e., absent from all tubes in the methodology employed), (ii) an average MPN of fewer than 50 coliforms per gram (10 or more pies), (iii) the absence of coagulase-positive staphylococci in 0.1-g portions, and (iv) an aerobic plate count of fewer than 25,000 per gram (geometric mean of 10 or more pies).     

Bacteriological Survey of the Blue Crab Industry

During sanitation inspections of 46 crabmeat processing plants on the Atlantic and Gulf Coasts, 487 samples of whole crabs immediately after cooking, cooked crabs after cooling, backed or washed (or both) crab bodies and whole crab claws, as well as 1,506 retail units of finished product were collected and analyzed bacteriologically. The 1,506 retail units (1-lb [373.24-g] cans) included 518 cans of regular (special) meat, 487 cans of claw meat, and 501 cans of lump meat. Statistical analyses showed that crabmeat from plants in Mississippi, Louisiana, and Texas had higher counts in 19 of 24 cases for the four bacteriological indices than crabmeat from plants located along the Atlantic Coast and the Gulf Coast of Florida. Aerobic plate counts of retail units collected from a previous day's production were significantly higher than those collected on the day of inspection. Regular crabmeat had consistently higher aerobic plate counts than claw or lump meat. When the product was handled expeditiously under good sanitary conditions, the bacteriological results were significantly better than the results from plants operating under poor sanitary conditions. Crabmeat produced in plants operating under good sanitary conditions had the following bacteriological content: (i) coliform organisms average most-probable-number values (geometric) of less than 20 per g; (ii) no Escherichia coli; (iii) coagulase-positive staphylococci average most-probable-number values (geometric) of less than 30 per g in 93% of the plants; (iv) aerobic plate count average values (geometric) of less than 100,000 per g in 93% of the plants, with the counts from 85% of these plants below 50,000 per g.     

Bacteriological Survey of Frozen Meat and Gravy Produced at Establishments Under Federal Inspection

During visits to 34 federally inspected establishments producing frozen meat and gravy, 541 production line samples and 535 finished product units were collected for bacteriological analyses. It was found that more than 70% of the sets of finished product (10 units/set) produced under good manufacturing practices had: (i) four or fewer coliform-positive units, (ii) two or fewer Escherichia coli-positive units, (iii) three or fewer Staphylococcus aureus-positive units, and (iv) an aerobic plate count of fewer than 50,000/g (geometric mean of 10 units). All finished product units were negative for salmonellae.     

Microbiological Evaluation of Cold-Water Shrimp (Pandalus borealis)

A bacteriological survey of the Maine shrimp industry was conducted to investigate the conditions associated with the production of frozen, raw, peeled shrimp. In-plant samples and finished product units were collected from seven plants. The most probable number of Escherichia coli, coliforms, and coagulase-positive staphylococci, as well as aerobic plate counts (APC), were determined. Freshly harvested shrimp collected from fishing vessels had an APC geometric mean of 510/g, and E. coli, coliforms, and coagulase-positive staphylococci were absent. Subsequent storage and insanitary practices during processing increased the APC and introduced coliforms. However, the low air temperatures (18 to 45 F) in the plants and the large volumes of cold water (34 F) used during processing inhibited significant bacterial buildup in the finished product.     

Changes in microbial population during fermentation of tropical freshwater fish viscera

Freshwater fish viscera (FV) was homogenized, mixed with 10% (w/w of FV) molasses and 0, 2 or 4% salt and allowed to ferment at ambient temperature (26·2°C) under microaerophilic conditions. The results revealed a reduction in total viable count and the number of spores, coliforms, Escherichia coli , staphylococci and enterococci and an increase in yeasts and moulds and lactic acid bacteria during fermentation. Coliforms and E. coli were found to be absent after 6 d and enterococci on 8th day. The presence of salt resulted in a marginally lower number of all organisms except yeasts, moulds and lactic acid bacteria. Inclusion of either 0·5% propionic acid, 0·3% calcium propionate or 0·1% sorbic acid suppressed growth of yeasts and moulds with propionic acid being the most effective. The study indicated that a microbiologically stable product could be prepared by ensiling fish viscera with 10% molasses and 0·5% propionic acid.     

Staphylococci in Competition: I. Growth of Naturally Occurring Mixed Populations in Precooked Frozen Foods during Defrost

In chicken pot pies, it was not possible to promote the growth of appreciable numbers of staphylococci under any condition of defrost. The pies spoiled under the same conditions, however. In macaroni and cheese dinners, tremendous numbers of saprophytic bacteria developed, but only after extended incubation at room temperature in which even spoilage was carried to an extreme. Under these conditions, staphylococci also multiplied vigorously. At the extreme temperature of 37 C, rank spoilage of such an advanced state as to render the product completely inedible was reached in less than 24 hr. Staphylococci grew very well under these extreme conditions.     

Coliform Behavior in Frozen Foods: I. Rapid Test for the Recovery of Escherichia coli from Frozen Foods

An assortment of 496 samples of frozen foods consisting of fish or marine products, variety types, and cream pie desserts were subjected to four parallel examinations for the recovery of Escherichia coli. The test procedures consisted of two low-temperature (35 C) and two high-temperature (44 C) presumptive tests, followed by an E C confirmatory test at 45.5 C. Of all the test methods examined, a single Lauryl Sulfate Tryptose (LST) presumptive test at 44 C gave best E. coli recovery (425). This recovery compared favorably with the lengthier Association of Official Analytical Chemists test with which only 420 E. coli cells were recovered. The LST (44 C) test saves much time, since it renders a follow-up 48-hr confirmatory test unnecessary. Moreover, since 96% of all the E. coli are recovered within 24 hr by LST (44 C), it is essentially a 24-hr test. The results of this study also confirmed earlier findings, in that it is possible to describe a specific coliform bacteriological test method by simple reproducible productivity ratios. E. coli recovery dilution data and coliform group behavior were also examined.     

Sanitary housing conditions modify the performance and behavioural response of weaned pigs to feed- and housing-related stressors

Pigs are confronted with changes in farming practices that may affect performance and animal well-being. The sanitary conditions of the farm can have an impact on the ability of pigs to adapt to these changes. This study aimed to analyse how weaned pigs respond to common farming practices of changes in diet and housing in terms of performance, health and behaviour, and how these responses are affected by the sanitary housing conditions, qualified here as good or poor. At weaning at 4 weeks of age, 20 piglets were assigned to 10 blocks of two littermates and each pig within a litter was randomly assigned to one of two sanitary conditions. Pigs were housed individually and received a starter diet. A diet change occurred on day 12 post weaning (starter to weaner diets) and pigs were transferred to the grower unit on day 33 post weaning and continued to receive the weaner diet. From 43 days post weaning, pigs were offered a grower diet and were vaccinated against swine influenza on day 47 and 61 post weaning. On the basis of this design, three post-weaning phases were identified: phase I from day 1 to 11 (post weaning), phase II from day 12 to 32 (after the diet change) and phase III from day 33 to 42 (after the housing change). Individual BW was measured every 3 days, and feed refusals and faecal scores were recorded on a daily basis. Behavioural observations were performed during 28 days by using the instantaneous scan sampling method. Individual blood samples were collected at the end of each phase to analyse the plasma concentration of haptoglobin and on day 68 post weaning to analyse the anti-influenza immunoglobulins G (IgG). Poor sanitary conditions resulted in a decrease in daily gain, feed intake and gain to feed ratio of, respectively, 11%, 5% and 7% (P < 0.05). Pigs in poor sanitary conditions had higher faecal scores (P < 0.05), tended to have higher plasma haptoglobin concentration in phase II (P = 0.06) and had a higher anti-influenza IgG titre (P = 0.11). The diet change affected performance and behavioural responses of pigs in poor but not in good sanitary conditions. Housing change resulted in a 30% decrease in growth and an increase in behaviour oriented towards exploration and excitement. The results of this study show an effect of sanitary conditions on the responses of pigs to a diet change, whereas those to a housing change were little affected by the sanitary conditions.     

Microbiological quality changes in the intestine of hybrid tilapia (Oreochromis niloticus x Oreochromis aureus) in fresh and frozen storage condition

AIMS: The goal of this study was to monitor the quantitative and qualitative bacterial flora in the intestine of hybrid tilapia in fresh fish and fish kept in frozen storage conditions for 1 year. METHODS AND RESULTS: Quantitative and qualitative analyses of the bacterial flora associated with the intestine of hybrid tilapia (Oreochromis niloticus x Oreochromis aureus) in fresh fish and fish kept in frozen storage conditions for 1 year were carried out. In fresh and frozen fish, aerobic plate count (APC) ranged from 1.6 +/- 1.2 x 10(8) to 1.5 +/- 0.9 x 10(5) CFU g(-1) in the intestine of tilapia collected from pond 1, 8.7 +/- 2.3 x 10(7) to 6.5 +/- 3.8 x 10(4) CFU g(-1) in the intestine of tilapia from pond 2, and 1.9 +/- 2.9 x 10(8) to 6.2 +/- 2.8 x 10(4) CFU g(-1) in the intestine of tilapia from pond 3. APC for all the groups of fish decreased c. 2-log cycles after 1 months frozen storage; thereafter, counts slowly declined during frozen storage for 1 year. Altogether, 16 bacterial genera were identified: Gram-negative rods (67%) dominated. Both in fresh and frozen conditions, four bacterial species viz. Shewanella putrefaciens, Corynebacterium urealyticum, Aeromonas hydrophila and Flavobacterium sp. were always present, with a prevalence of 10% in most cases. Shewanella putrefaciens was the most dominant organism (15% of the total isolates) throughout the studied period. During frozen storage some of the bacteria were not recovered, but most of the bacteria survived after prolonged freezing. CONCLUSIONS: This study describes the aerobic heterotrophic microflora found in the intestine of fresh and frozen tilapia. The unique aspect of this study concerns the data revealing the micro-organisms, which are viable after prolonged freezing. Contamination of edible portions of fish could originate from gastrointestinal sources. SIGNIFICANCE AND IMPACT OF THE STUDY: The present results may enhance knowledge in controlling the storage life of fish, and fish product quality. Bacterial activity is by far the most important factor influencing fish quality, so bacterial numbers can be used as an index of quality. Storage of frozen tilapia without evisceration could be avoided.     

EVALUATION OF THE PETRIFILMTM AND REDIGELTM AS RAPID METHODS TO THE STANDARD PLATE COUNT METHOD FOR THE ENUMERATION OF PROCESSED MEATS AND ENVIRONMENTAL SAMPLES

Aerobic plate counts (APC) are used by the food industry to help determine the sanitary state of equipment and bacterial load of finished product. Conventional aerobic count methods for processed meats require approximately 72 h incubation before results are available. Food processing plants with in-house analytical laboratories require methods that are simple, reliable and rapid. New test methods using Petrifilm^TM (PF) and Redigel^TM (RG) have been developed for determining APC. These methods are faster and easier than culturing in a standard agar medium. A variety of raw, cooked; cured and noncured meat products, cooling brine and environmental surface swabs were collected and analyzed for APC using PF, RG and plate count agar (PCA). Data analyses from over 200 samples indicated that the sensitivity of rapid testing methods for aerobic bacteria can vary depending on sample type.     

Hyperosmotic infiltration: immunological demonstration of infiltrating bacteria in brown trout, Salmo trutta L.

Brown trout were subjected to the hyperosmotic infiltration method of bacterial immunization. The route of entry of E. coli into fish was observed by the immunoperoxidase technique on instantly frozen tissue to be through the gills. The passage of bacteria directly into the lateral line system of the fish was not observed.     

Microbiological quality of frozen breaded fish and shellfish products.

A survey was made of the microbiological quality of seven frozen, breaded, precooked fish and shellfish products and of frozen, breaded, uncooked shrimp at the retail level. Geometric mean aerobic plate counts per gram (and number of units examined) were as follows: fish sticks, 8,300 (1,539); fish cakes, 5,600 (1,378); crab cakes, 4,900 (1,226); scallops, 1,700 (1,392); clams, 450 (1,384); haddock, 15,000 (1,306); fish in fish and chips dinner, 7,200 (1,485); and uncooked shrimp, 220,000 (1,462). Geometric mean coliform, Escherichia coli, and Staphylococcus aureus counts for all eight products ranged from 1 to 10/g.     

Protein Linewidth and Solvent Dynamics in Frozen Solution NMR

Solid-state NMR of proteins in frozen aqueous solution is a potentially powerful technique in structural biology, especially if it is combined with dynamic nuclear polarization signal enhancement strategies. One concern regarding NMR studies of frozen solution protein samples at low temperatures is that they may have poor linewidths, thus preventing high-resolution studies. To learn more about how the solvent shell composition and temperature affects the protein linewidth, we recorded ¹H, ²H, and ¹³C spectra of ubiquitin in frozen water and frozen glycerol-water solutions at different temperatures. We found that the ¹³C protein linewidths generally increase with decreasing temperature. This line broadening was found to be inhomogeneous and independent of proton decoupling. In pure water, we observe an abrupt line broadening with the freezing of the bulk solvent, followed by continuous line broadening at lower temperatures. In frozen glycerol-water, we did not observe an abrupt line broadening and the NMR lines were generally narrower than for pure water at the same temperature. ¹H and ²H measurements characterizing the dynamics of water that is in exchange with the protein showed that the ¹³C line broadening is relatively independent from the arrest of isotropic water motions.     

Elucidation of Listeria monocytogenes contamination routes in cold-smoked salmon processing plants detected by DNA-based typing methods

The contamination routes of Listeria monocytogenes in cold-smoked salmon processing plants were investigated by analyzing 3,585 samples from products (produced in 1995, 1996, 1998, and 1999) and processing environments (samples obtained in 1998 and 1999) of two Danish smokehouses. The level of product contamination in plant I varied from 31 to 85%, and no L. monocytogenes was found on raw fish (30 fish were sampled). In plant II, the levels of both raw fish and product contamination varied from 0 to 25% (16 of 185 raw fish samples and 59 of 1,000 product samples were positive for L. monocytogenes). A total of 429 strains of L. monocytogenes were subsequently compared by random amplified polymorphic DNA (RAPD) profiling, and 55 different RAPD types were found. The RAPD types detected on the products were identical to types found on the processing equipment and in the processing environment, suggesting that contamination of the final product (cold-smoked salmon) in both plants (but primarily in plant I) was due to contamination during processing rather than to contamination from raw fish. However, the possibility that raw fish was an important source of contamination of the processing equipment and environment could not be excluded. Contamination of the product occurred in specific areas (the brining and slicing areas). In plant I, the same RAPD type (RAPD type 12) was found over a 4-year period, indicating that an established in-house flora persisted and was not eliminated by routine hygienic procedures. In plant II, where the prevalence of L. monocytogenes was much lower, no RAPD type persisted over long periods of time, and several different L. monocytogenes RAPD types were isolated. This indicates that persistent strains may be avoided by rigorous cleaning and sanitation; however, due to the ubiquitous nature of the organism, sporadic contamination occurred. A subset of strains was also typed by using pulsed-field gel electrophoresis and amplified fragment length polymorphism profiling, and these methods confirmed the type division obtained by RAPD profiling.     

Elucidation of Listeria monocytogenes Contamination Routes in Cold-Smoked Salmon Processing Plants Detected by DNA-Based Typing Methods

The contamination routes of Listeria monocytogenes in cold-smoked salmon processing plants were investigated by analyzing 3,585 samples from products (produced in 1995, 1996, 1998, and 1999) and processing environments (samples obtained in 1998 and 1999) of two Danish smokehouses. The level of product contamination in plant I varied from 31 to 85%, and no L. monocytogenes was found on raw fish (30 fish were sampled). In plant II, the levels of both raw fish and product contamination varied from 0 to 25% (16 of 185 raw fish samples and 59 of 1,000 product samples were positive for L. monocytogenes). A total of 429 strains of L. monocytogenes were subsequently compared by random amplified polymorphic DNA (RAPD) profiling, and 55 different RAPD types were found. The RAPD types detected on the products were identical to types found on the processing equipment and in the processing environment, suggesting that contamination of the final product (cold-smoked salmon) in both plants (but primarily in plant I) was due to contamination during processing rather than to contamination from raw fish. However, the possibility that raw fish was an important source of contamination of the processing equipment and environment could not be excluded. Contamination of the product occurred in specific areas (the brining and slicing areas). In plant I, the same RAPD type (RAPD type 12) was found over a 4-year period, indicating that an established in-house flora persisted and was not eliminated by routine hygienic procedures. In plant II, where the prevalence of L. monocytogenes was much lower, no RAPD type persisted over long periods of time, and several different L. monocytogenes RAPD types were isolated. This indicates that persistent strains may be avoided by rigorous cleaning and sanitation; however, due to the ubiquitous nature of the organism, sporadic contamination occurred. A subset of strains was also typed by using pulsed-field gel electrophoresis and amplified fragment length polymorphism profiling, and these methods confirmed the type division obtained by RAPD profiling.     

Capsules for Frozen Sectioning of Small Specimens

It was necessary to make sections of small unfixed specimens which had been frozen while still immersed in their normal culture medium. The principal difficulty stemmed from the poor sectioning quality of the frozen culture medium. A capsule is described which has a narrow well in which the tissue specimen fits snugly within a small amount of culture medium. After freezing, the whole capsule is sectioned and the resulting sections, being nearly devoid of culture medium, are of good quality.     

Molecular studies on the ecology of Listeria monocytogenes in the smoked fish processing industry

We have applied molecular approaches, including PCR-based detection strategies and DNA fingerprinting methods, to study the ecology of Listeria monocytogenes in food processing environments. A total of 531 samples, including raw fish, fish during the cold-smoking process, finished product, and environmental samples, were collected from three smoked fish processing facilities during five visits to each facility. A total of 95 (17.9%) of the samples tested positive for L. monocytogenes using a commercial PCR system (BAX for Screening/Listeria monocytogenes), including 57 (27.7%) environmental samples (n = 206), 8 (7.8%) raw material samples (n = 102), 23 (18.1%) samples from fish in various stages of processing(n = 127), and 7 (7.3%) finished product samples (n = 96). L. monocytogenes was isolated from 85 samples (16.0%) using culture methods. Used in conjunction with a 48-h enrichment in Listeria Enrichment Broth, the PCR system had a sensitivity of 91.8% and a specificity of 96.2%. To track the origin and spread of L. monocytogenes, isolates were fingerprinted by automated ribotyping. Fifteen different ribotypes were identified among 85 isolates tested. Ribotyping data established possible contamination patterns, implicating raw materials and the processing environment as potential sources of finished product contamination. Analysis of the distribution of ribotypes revealed that each processing facility had a unique contamination pattern and that specific ribotypes persisted in the environments of two facilities over time (P < or = 0.0006). We conclude that application of molecular approaches can provide critical information on the ecology of different L. monocytogenes strains in food processing environments. This information can be used to develop practical recommendations for improved control of this important food-borne pathogen in the food industry.     

Salmonellae in Fish Meal Plants: Relative Amounts of Contamination at Various Stages of Processing and a Method of Control

Previous studies have shown that Menhaden fish meal, a common ingredient of animal feeds, is frequently contaminated with salmonellae. Animals that eat contaminated feed may become infected. If they, in turn, are eaten by humans, they may be a means by which salmonellae are introduced into the human population. Epidemiological studies of the fish-meal industry were carried out to determine the sources of salmonellae in fish meal and the factors affecting the persistence and survival of salmonellae during the processing of fish meal. Examination of 190 fish immediately after they came from the Gulf of Mexico revealed no salmonellae, but salmonellae were frequently isolated from samples of fish taken from the boats when they arrived at the plants. Salmonellae were also frequently isolated from dockside water at each of the plants. Approximately 50% of the samples taken in the raw fish processing areas were contaminated with salmonellae. The percentage of samples yielding salmonellae decreased progressively through the various sequences of processing, but more than 15% of the samples taken from the finished products were also positive. Salmonellae were isolated from the raw area of the plant most frequently while the plant was operating and less frequently when the plant was idle, whereas in the processing area of the plant the reverse was true. Salmonellae appeared to survive and multiply in the processing area of the plant while the plant was idle, which resulted in contamination of the first portion of each day's production. Salmonellae in the processed fish meal were reduced to nondetectable levels by reprocessing the first 45 min of each day's production.