Bacteriological Survey of the Frozen Prepared Foods Industry: IV. Frozen Breaded Fish

During sanitation-inspections of 23 breaded-fish processors, 573 finished product units and 604 line samples were collected and analyzed bacteriologically. Data for finished product units processed on the same data are presented in groups; most of the groups consisted of 10 units. Groups of frozen, raw-breaded fish produced under good and poor conditions of sanitation were studied. No more than 20% of the units in each group produced under good conditions of sanitation were positive for Escherichia coli or coagulase-positive staphylococci. Groups of frozen, fried-breaded fish produced under good and poor sanitary conditions were also studied. No more than 10% of the units in each good-sanitation group were positive for E. coli or coagulase-positive staphylococci. However, approximately 30% of the groups processed under poor sanitary conditions also did not exceed these viable bacterial counts because of the lethal effect of the terminal fry to which the product is subjected. It was found that line samples collected from the processing lines reflected the sanitary conditions of the plant and provided bacteriological support for inspectional evidence of plant insanitation.     

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 Fresh Pork Sausage Produced at Establishments Under Federal Inspection

At the time of manufacture, 75% of 67 sets of finished fresh pork sausage collected in 44 plants had aerobic plate counts in the range of 500,000 or fewer/g; 88% contained 100 or fewer E. coli/g; and 75% contained 100 or fewer S. aureus/g (geometric means of 10 samples). Salmonellae were isolated from 28% of 529 samples of pork trimmings used for sausage, and from 28% of 560 finished sausage samples. Semiquantitative analysis revealed that salmonellae were at low levels; more than 80% of the salmonellae-positive samples were positive only in 25-g portions (negative in 1.0- and 0.1-g portions).     

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.     

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.     

The distribution and frequency of microsatellite loci in Drosophila melanogaster

We report the results of a comprehensive search of Drosophila melanogaster DNA sequences in GenBank for di-, tri-, and tetranucleotide repeats of more than four repeat units, and a DNA library screen for dinucleotide repeats. Dinucleotide repeats are more abundant (66%) than tri- (30%) or tetranucleotide (4%) repeats. We estimate that 1917 dinucleotide repeats with 10 or more repeat units are present in the euchromatic D. melanogaster genome and, on average, they occur once every 60 kb. Relative to many other animals, dinucleotide repeats in D. melanogaster are short. Tri- and tetranucleotide repeats have even fewer repeat units on average than dinucleotide repeats. Our WorldWide Web site (http://www.bio.cornell.edu/genetics/aquadro/aquadro.html) posts the complete list of 1298 microsatellites (≥ five repeat units) identified from the GenBank search. We also summarize assay conditions for 70 D. melanogaster microsatellites characterized in previous studies and an additional 56 newly characterized markers.     

Ovipositional Synergists and Artificial Diets for Trichogramma australicum Girault (Hymenoptera: Trichogrammatidae)

We studied ovipositional synergists and artificial diets for rearing Trichogramma australicum. Artificial "diet A" included 40% haemolymph of Helicoverpa armigera (Hübner) final instar larvae, 30% of a 10% malt solution in deionised water, 20% chicken egg yolk and 10% Neisenheimer's salt solution with 76 units Penicillin and 76 units Streptomycin/ml diet. Artificial "diet B" was identical except Grace's insect medium® replaced Neisenheimer's salt solution. the number of T. australicum larvae in artificial eggs filled with diet A and diet B was not significantly different (P > 0.05). Significantly fewer (P < 0.05) larvae developed to pupae and adults in artificial eggs filled with diet A. Quantity and quality of artificial diet affected the mortality of T. australicum larvae reared in vitro. Ovipositional synergists included 10% gelatine solution, 10% polyvinyl alcohol solution or 1% agar solution in deionised water. Synergist test-solutions were individually smeared on the external surface of artificial eggs (hemispherical depressions in plastic membrane). Eggs laid and number of T. australicum larvae produced were significantly higher in artificial eggs smeared with gelatine than artificial eggs smeared with polyvinyl alcohol, agar or non-smeared (control) eggs.     

Synchronization of larval emergence in winter moth (Operophtera brumata L.) and budburst in pedunculate oak (Quercus robur L.) under simulated climate change

1. The hypothesis that a 3 °C elevation in temperature and doubled CO₂ concentration would have no effect on the synchronization of winter moth egg hatch with budburst in oak was tested by comparing the separate and interactive effects of ambient and elevated (+ 3 °C) temperature and ambient and elevated (doubled to 340 p.p.m.) CO₂ in eight experimental Solardomes. In addition, an outdoor control was compared with the ambient temperature/CO₂ treatment combination.
2. Elevated temperature accelerated darkening (preceding egg hatch by about 5–10 days) and hatching of eggs developing off the trees; elevated CO₂ had no effect. The same effects were observed in eggs developing on the trees.
3. Within treatments, date of egg hatch was the same on trees with early or late budburst.
4. Egg darkening and budburst were closely synchronized at both ambient and elevated temperatures.
5. Both eggs and trees required fewer cumulative heat units (day degrees > 4 °C), for hatching and budburst, respectively, at ambient than elevated temperatures. The requirements in the outdoor control treatment were similar to those in the ambient Solardome treatment.
6. Egg hatch between 10 and 25 °C, on a temperature gradient in the laboratory, required a constant number of heat units; fewer were required below 10 °C.
7. Elevated temperatures, in the Solardomes and the field, delayed adult emergence from the pupae.
8. The results suggest that a general increase in temperature with climatic change would not affect the closeness of the synchronization between egg hatch of winter moth and budburst of oak.     

Elimination of viruses and indicator bacteria at each step of treatment during preparation of drinking water at seven water treatment plants

Seven drinking water treatment plants were sampled twice a month for 12 months to evaluate the removal of indicator bacteria and cytopathogenic enteric viruses. Samples were obtained at each level of treatment: raw water, postchlorination, postsedimentation, postfiltration, postozonation, and finished (tap) water. Raw water quality was usually poor, with total coliform counts exceeding 105 to 106 CFU/liter and the average virus count in raw water of 3.3 most probable number of cytopathogenic units (MPNCU)/liter; several samples contained more than 100 MPNCU/liter. All plants distributed finished water that was essentially free of indicator bacteria as judged by analysis of 1 liter for total coliforms, fecal coliforms, fecal streptococci, coagulase-positive staphylococci, and Pseudomonas aeruginosa. The total plate counts at 20 and 35 degrees C were also evaluated as a measure of the total microbial population and were usually very low. Viruses were detected in 7% (11 of 155) of the finished water samples (1,000 liters) at an average density of 0.0006 MPNCU/liter the highest virus density measured being 0.2 MPNCU/liter. The average cumulative virus reduction was 95.15% after sedimentation and 99.97% after filtration and did not significantly decrease after ozonation or final chlorination. The viruses isolated from treated waters were all enteroviruses: poliovirus types 1, 2, and 3, coxsackievirus types B3, B4, and B5, echovirus type 7, and untyped picornaviruses.     

Elimination of viruses and indicator bacteria at each step of treatment during preparation of drinking water at seven water treatment plants.

Seven drinking water treatment plants were sampled twice a month for 12 months to evaluate the removal of indicator bacteria and cytopathogenic enteric viruses. Samples were obtained at each level of treatment: raw water, postchlorination, postsedimentation, postfiltration, postozonation, and finished (tap) water. Raw water quality was usually poor, with total coliform counts exceeding 105 to 106 CFU/liter and the average virus count in raw water of 3.3 most probable number of cytopathogenic units (MPNCU)/liter; several samples contained more than 100 MPNCU/liter. All plants distributed finished water that was essentially free of indicator bacteria as judged by analysis of 1 liter for total coliforms, fecal coliforms, fecal streptococci, coagulase-positive staphylococci, and Pseudomonas aeruginosa. The total plate counts at 20 and 35 degrees C were also evaluated as a measure of the total microbial population and were usually very low. Viruses were detected in 7% (11 of 155) of the finished water samples (1,000 liters) at an average density of 0.0006 MPNCU/liter the highest virus density measured being 0.2 MPNCU/liter. The average cumulative virus reduction was 95.15% after sedimentation and 99.97% after filtration and did not significantly decrease after ozonation or final chlorination. The viruses isolated from treated waters were all enteroviruses: poliovirus types 1, 2, and 3, coxsackievirus types B3, B4, and B5, echovirus type 7, and untyped picornaviruses.     

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.     

Association between transfusion of whole blood and recurrence of cancer.

Transfusion affects the immune response to renal transplantation and may be associated with recurrence of various human neoplasms. Data from patients with colonic, rectal, cervical, and prostate tumours showed an association between transfusion of any amount of whole blood or larger amounts of red blood cells at the time of surgery and later recurrence of cancer. Recipients of one unit of whole blood had a significantly higher incidence of recurrence (45%) than recipients of a single unit of red cells (12%) (p = 0.03). Recipients of two units of whole blood also had a higher rate of recurrence (52%) than those receiving two units of red cells (23%) (p = 0.03). Recipients of any amount of whole blood had similar recurrence rates (38-52%). Recipients of four or more units of red blood cells had a higher rate of recurrence (55%) than those receiving three or fewer units of red blood cells (20%) (p = 0.005). Mortality due to cancer in patients receiving three or fewer units of red blood cells (2%) was similar to that in patients who did not have transfusions (7%) and significantly lower than that observed in patients receiving three or fewer units of whole blood (20%) (p = 0.003). A proportional hazards risk analysis showed that transfusion of any whole blood or more than three units of red blood cells was significantly associated with earlier recurrence and death due to cancer. These data support an association between transfusion and recurrence of cancer. They also suggest that some factor present in greater amounts in whole blood, such as plasma, may contribute to the increased risk of recurrence in patients who have undergone transfusion. Until the questions raised by retrospective studies of cancer recurrence and transfusion can be answered by prospective interventional trials with washed red blood cells, red blood cells should be transfused to patients with cancer in preference to whole blood when clinically feasible.     

IMP-GMP 5'-nucleotidase in reptiles: occurrence and tissue distribution in a crocodile and three species of lizards

IMP-hydrolyzing activity (which is reactive with goose anti-pig lung IMP-GMP 5'-nucleotidase (c-N-II: EC.3.1.3.5) serum) was detected in extracts from several tissues (liver, heart, kidney, spleen, stomach, lung and skeletal muscle) from constitutively uricotelic reptiles: a crocodile (Crocodylus siamensis), and three species of lizard (Furcifer oustaleti, Tupinambis rufescens and Varanus gouldi). The activities were markedly high in the livers: 3.0 units/g in the crocodile and 1.4-2.9 units/g in the lizards. These were similar to those previously reported for the livers from chicken and snakes (also constitutively uricotelic), and 4- to 10-fold higher than those in ammoniotelic or ureotelic vertebrates. These findings suggest that the high activity of IMP-GMP 5'-nucleotidase in the liver is a feature of constitutive uricotelism, and that the enzyme may participate in the production of uric acid as an end product of amino acid catabolism.     

Evaluation of an anti-inflammatory factor derived from hyperimmunized cows

An anti-inflammatory factor isolated from milk of hyperimmunized cows was analyzed in vitro and in vivo. Macrophages collected from lacteal secretions of a unimmunized nonlactating cow showed increased ability to kill phagocytosed Staphylococcus aureus when incubated with the anti-inflammatory factor. Mice injected intraperitoneally with 10 mg/kg of anti-inflammatory factor demonstrated an increased LD50 to S. aureus when challenged intraperitoneally. Injected mice also demonstrated significantly (P less than 0.05) less mammary inflammation and involution and increased clearance of S. aureus when challenged intramammarily. Quantitative histologic analysis of mammary tissues from mice injected with anti-inflammatory factor demonstrated significantly (P less than 0.05) more lumen, less interalveolar connective tissue, and less leukocytic infiltration compared with control mice. Mammary glands of mice injected with anti-inflammatory factor and challenged with S. aureus also contained fewer colony-forming units than control mice. The product appeared to exert its effect on the nonspecific defense system via modulation of leukocyte function.     

Bacteriological quality and shelf life of ground beef.

The bacteriological quality of unfrozen raw ground beef was evaluated after 0, 3, 6, 9, 12, 15, and 18 days of storage at 29 +/- 1 F (-1.7 +/- 0.6 C). At the time of fabrication, all of the ground beef samples contained 10(6) or fewer total aerobic and psychrotrophic bacteria/g; 81% contained 100 or fewer coliforms/g; 94% contained 100 or fewer Escherichia coli/g; and all of the samples contained 100 or fewer coagulase-positive Staphylococcus aureus and Clostridium perfringens/g. Total aerobic and psychrotrophic bacteria increased by 1 log between 3 and 18 days of storage. Coliform and E. coli counts decreased during storage, whereas coagulase-positive S. aureus and C. perfringens counts did not change significantly. These data indicate that meat processors, wholesalers, and retailers could improve the bacteriological quality and prolong the shelf life of ground beef packaged in oxygen-impermeable film if the temperature of product never exceeded 29 +/- 1 F (-1.7 +/- 0.6 C).     

Comparison of automated culture systems with a CFR/USP-compliant method for sterility testing of cell-therapy products

BACKGROUND: Although widely used, commercially available automated culture methods are not US Food and Drug Administration-approved for sterility testing of cell-therapy products. For cell-therapy products regulated under Section 351 of the Public Health Service Act, sterility testing must be performed by the methods described in 21 CFR 610.12 and USP <71> (CFR/USP method), or by methods demonstrated to be equivalent. METHODS: Two automated methods, BacT/Alert (BTA; bioMerieux) and Bactec (Becton Dickinson), were compared with the CFR/USP method. Representative mononuclear cell (MNC) products were formulated using six different product media. MNC product aliquots containing 10-50 x 10(6) cells in a 0.5 mL volume were seeded with organisms, and cultured for 14 days in aerobic and anaerobic bottles of each system. Ten different organisms at target concentrations of 10 and 50 colony-forming units (CFU) per bottle were tested. RESULTS: Positives were detected in a mean (range) of 72% (7-100%) of cultures for CFR/USP, 82% (0-100%) for BTA, and 93% (57-100%) for Bactec. For nine of the 10 organisms tested, overall detection rates for BTA and Bactec were equivalent to or higher than CFR/USP. Of the six product media tested, detection of organisms was impaired only by the medium containing multiple antibiotics: this occurred in all three systems. Both BTA and Bactec had shorter times to detection than the CFR/USP method, with overall means (ranges) of 87 (24-264) h for CFR/USP, 24 (12-54) h for BTA, and 33 (12-80) h for Bactec. Detection occurred consistently within 7 days for both BTA and Bactec, but not for CFR/USP. DISCUSSION: Both BTA and Bactec are superior to the CFR/USP method for overall detection and time to detection of organisms in MNC products suspended in commonly used media. These data support general use of either BTA or Bactec for sterility testing of a variety of cell-therapy products, and suggest that a 7-day culture period is sufficient to detect clinically relevant organisms. These results confirm the need for bacteriostasis and fungistasis testing of antibiotic-containing products, even when antibiotic-binding substances are used.     

Bacteriological quality and shelf life of ground beef.

The bacteriological quality of unfrozen raw ground beef was evaluated after 0, 3, 6, 9, 12, 15, and 18 days of storage at 29 +/- 1 F (-1.7 +/- 0.6 C). At the time of fabrication, all of the ground beef samples contained 10(6) or fewer total aerobic and psychrotrophic bacteria/g; 81% contained 100 or fewer coliforms/g; 94% contained 100 or fewer Escherichia coli/g; and all of the samples contained 100 or fewer coagulase-positive Staphylococcus aureus and Clostridium perfringens/g. Total aerobic and psychrotrophic bacteria increased by 1 log between 3 and 18 days of storage. Coliform and E. coli counts decreased during storage, whereas coagulase-positive S. aureus and C. perfringens counts did not change significantly. These data indicate that meat processors, wholesalers, and retailers could improve the bacteriological quality and prolong the shelf life of ground beef packaged in oxygen-impermeable film if the temperature of product never exceeded 29 +/- 1 F (-1.7 +/- 0.6 C).     

Effects of lead on luteal function in rhesus monkeys

Exposure to lead in the workplace or home environment has been implicated as a cause of decreased fertility in women. In a previous study, as part of our effort to determine effects of lead in primates, female rhesus monkeys were exposed to lead acetate in drinking water (n = 10) or provided water with no added lead (n = 7) for 33 mo. Lead was administered at levels between 2 and 8 mg/kg/day, with doses adjusted to keep blood lead values near a target of 70 micrograms/dl (observed mean +/- SEM = 68.9 +/- 6.54 micrograms/dl). Blood lead concentrations in control animals were less than 10 micrograms/dl. No significant differences were detected between control and experimental animals in body weight, hematocrit, or general health. Female monkeys receiving lead exhibited longer and more variable menstrual cycles and shorter menstrual flow. In the present study, circulating amounts of progesterone (P4) were determined to evaluate luteal function during the final 7 mo of treatment with lead. Several characteristics were altered as a result of lead treatment: circulating amounts of P4 were reduced as indicated by relative units of area under the concentration-time curve, maximal amounts of P4 were reduced, and P4 levels were greater than 1 ng/ml on fewer days. There were no significant differences between groups in mean percent of anovulatory cycles. Therefore, although chronic treatment with the levels of lead used in this study did not prevent ovulation, luteal function was suppressed. These results extend previous observations of adverse effects of lead on ovarian activity and fertility in monkeys.