Search

Toxocara canis is a zoonotic nematode parasite that can be transmitted to humans by food or water contaminated with T. canis eggs from infected dog feces. High-pressure processing (HPP) is a useful alternative to thermal treatments to eliminate pathogens from foods. Most of the research on HPP has focused on prokaryotes, but little is known about its effects on eukaryotic organisms. We evaluated the ability of HPP to affect embryonation of T. canis eggs to test the hypothesis that HPP treatment can delay development of T. canis eggs. Efficacy of HPP was determined by using an embryonation assay on T. canis eggs from naturally infected puppies. For each treatment, 2500 T. canis eggs in tap water were placed in sealable plastic bags and subjected to 138-400 megapascals (MPa; 1 MPa=10 atm=147 psi) for 60 s in a commercial HPP unit. We found that treatment with 300 or 400 MPa for 60 s killed 100% of eggs using embryonation as the standard. Treatment with 250, 241, and 207 MPa was less effective and killed 80%, 56%, and 8% of eggs, respectively. Results from this study suggest that HPP may be a useful treatment to protect foods from T. canis contamination.

The model ordinance in the National Shellfish Sanitation Program's Guide for the Control of Molluscan Shellfish was initially established for oysters; however, the clam industry also follows the protocol. Rapid cooling during periods when the growing waters exceed 80 °F (26.7 °C) results in cold shock, which causes unacceptable mortalities in clams. The clam industry was looking for a procedure to lower the clams to the standard temperature while minimizing shell shock mortalities during the warm summer months. Three tempering treatments were examined, and total aerobic plate counts (APCs) and most-probable-number (MPN) counts of Vibrio, V. parahaemolyticus, and fecal coliforms were enumerated. In treatment 1 (control), clams were harvested, held for 5 h at 90 °F (32.2 °C), and then moved to 45 °F (7.2 °C) for storage. In treatment 2, clams were harvested and held for 5 h at 90 °F (32.2 °C), followed by 12 h at 65 °F (18.3 °C) and 12 h at 55 °F (12.8 °C), and then were moved to 45 °F (7.2 °C) for long-term storage. In treatment 3, clams were harvested and held for 5 h at 90 °F (32.2 °C), followed by 24 h at 55 °F (12.8 °C) before being moved to 45 °F (7.2 °C) for long-term storage. Three replicate trials were performed with triplicate analyses during late June through early to mid-August. The current National Shellfish Sanitation Program standard is treatment 1; it contained statistically (P ≤ 0.05) higher total APCs than treatments 2 and 3 throughout the 21-day storage period. APCs ranged from 2.3 × 10(4) immediately after harvest to 2.7 × 10(6), 1.6 × 10(5), and 4.8 × 10(5) for treatments 1, 2, and 3, respectively, after 14 days of storage. A statistical analysis showed that treatments 2 and 3 had significantly lower total MPN per gram Vibrio than treatment 1 on day 7 but were equal to treatment 1 on days 1 and 14. MPN per gram for V. parahaemolyticus was statistically lower in treatments 2 and 3 than in treatment 1 on storage days 1 and 7. However, on day 14, treatment 3 was significantly lower than treatments 1 and 2. There was no statistical difference for fecal coliforms. The greatest mortality occurred in treatment 1 (87.4%), followed by treatment 2 (83.3%) and treatment 3 (66.0%). The outcome of this research clearly shows that treatments 2 and 3 can cool clams to a temperature of 45 °F (7.2 °C) without compromising quality or safety and can reduce the number of dead clams introduced into the marketplace.

High hydrostatic pressure processing (HPP) is an effective non-thermal treatment used to inactivate pathogens from a variety of food and food products. It has been extensively examined using prokaryotic organisms and protozoan's but has had limited study on metazoans. Treatment using HPP has been shown to be effective in inactivating nematode larvae in food and preventing embryonation of Ascaris suum eggs. We conducted experiments using eggs of the canine whipworm Trichuris vulpis collected from naturally infected dogs and A. suum eggs from naturally infected pigs. We observed a delay in development of eggs of T. vulpis in a preliminary experiment and conducted 2 experiments to test the hypothesis that appropriate HPP levels can induce a delay in embryonation of nematode eggs. In experiment 1, nonembryonated T. vulpis eggs in tap water were packaged in sealable bags and exposed to 138-600 megapascals (MPa; 1 MPa=10 atm=147 psi) for 60s in a commercial HPP unit. In a second experiment, nonembryonated eggs of A. suum were exposed to 138-600 MPa and treated for 60s in the same commercial HPP unit. Embyronation of T. vulpis eggs was delayed by 4 and 5 days for eggs treated with 207 and 241 MPa but eventually eggs developed and the numbers of embryonated eggs was similar to controls on day 55. Embryonation of T. vulpis eggs treated with 345 or 350 MPa was delayed by 9 days and never reached more than 5% of eggs embryonated. On day 55 post treatment, 95% of control nontreated T. vulpis eggs were embryonated, 100-65% of eggs treated with 138-276 MPa were embryonated, a maximum of 5% of eggs treated with 345-350 MPa were embryonated, and 0% of eggs treated with ≥ 400 MPa were embryonated. T. vulpis eggs treated with ≥ 400 MPa did not undergo cell division. Embryrnation of A. suum eggs was delayed by 4, 10, and 16 days for eggs treated with 207, 241, and 250MPa, respectively, compared to nontreated control eggs. A. suum eggs treated with 207 MPa eventually embryonated to similar % embryonation values as controls and 138 MPa treated eggs but eggs treated with 241 or 250 MPa were always <5% embryonated. A. suum eggs treated with ≥ 300 MPa did not undergo cell division. On the final day of examination at day 56 after treatment, the % of embryonated eggs was 92% nontreated controls, 94% treated with 138 MPa, 84% treated with 207 MPa, 2% treated with 241 or 250 MPa, and 0% treated with 276, 200, 345, 400, or 414 MPa, respectively., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Contamination of oysters with human noroviruses (HuNoV) constitutes a human health risk and may lead to severe economic losses in the shellfish industry. There is a need to identify a technology that can inactivate HuNoV in oysters. In this study, we conducted a randomized, double-blinded clinical trial to assess the effect of high hydrostatic pressure processing (HPP) on Norwalk virus (HuNoV genogroup I.1) inactivation in virus-seeded oysters ingested by subjects. Forty-four healthy, positive-secretor adults were divided into three study phases. Subjects in each phase were randomized into control and intervention groups. Subjects received Norwalk virus (8FIIb, 1.0 × 10(4) genomic equivalent copies) in artificially seeded oysters with or without HPP treatment (400 MPa at 25°C, 600 MPa at 6°C, or 400 MPa at 6°C for 5 min). HPP at 600 MPa, but not 400 MPa (at 6° or 25°C), completely inactivated HuNoV in seeded oysters and resulted in no HuNoV infection among these subjects, as determined by reverse transcription-PCR detection of HuNoV RNA in subjects' stool or vomitus samples. Interestingly, a white blood cell (granulocyte) shift was identified in 92% of the infected subjects and was significantly associated with infection (P = 0.0014). In summary, these data suggest that HPP is effective at inactivating HuNoV in contaminated whole oysters and suggest a potential intervention to inactivate infectious HuNoV in oysters for the commercial shellfish industry.

High hydrostatic pressure processing (HPP) is an effective non-thermal treatment to remove pathogens from a variety of food and food products. It has been extensively examined using prokaryotic organisms but has had limited study on eukaryotic organisms. Treatment using HPP has been shown to be effective in inactivating nematode larvae in food and Ascaris suum eggs. Nothing is known on the efficacy of HPP on tapeworm cysts or eggs. Eggs of important zoonotic tapeworms including Echinococcus and Taenia spp. can potentially contaminate water and food intended for human consumption. The present study examined the efficacy of HPP on the viability of Hymenolepis diminuta eggs. Efficacy of HPP treatment was measured using an egg hatch assay in two experiments. One thousand unhatched H. diminuta eggs in Hanks balanced salt solution were packaged in sealable bags and exposed to 100-600megapascals (MPa; 1MPa=10atm=147psi) for 60s in a commercial HPP unit. Positive (no HPP) and negative (No HPP but frozen/thawed) controls were examined in each experiment. None of the HPP untreated and frozen eggs (negative controls) were able to hatch or exclude trypan blue when placed in the hatching solution in experiment 1 or 2. HPP untreated and nonfrozen eggs (positive controls) hatched and excluded trypan blue; 75% were positive in experiment 1 and 80% were positive in experiment 2. No hatched eggs were observed when they were exposed to 300-600MPa for 60s. Treatment at 400MPa and above caused rupturing of the oncosphere. Results from this study indicate that HPP is a possible method to inactivate tapeworm eggs and that the susceptibility of tapeworm eggs to HPP is similar to or greater than that of nematode eggs or tissue larvae., (Copyright © 2010 Elsevier B.V. All rights reserved.)

We have previously demonstrated that high pressure processing (HPP) is effective in preventing in vitro replication of murine norovirus strain 1 (MNV-1), a human norovirus surrogate, in a monocyte cell line following extraction from MNV-1-contaminated oysters. In the present study, the efficacy of HPP to prevent in vivo replication within mice fed HPP-treated MNV-1-seeded oyster extracts was evaluated. Oyster homogenate extracts seeded with MNV-1 were given 5-min, 400-MPa (58,016-psi) treatments and orally gavaged into immunodeficient (STAT-1(-/-)) female mice. Mice orally gavaged with HPP-treated MNV-1 showed significant (P ≤ 0.05) weight loss leading to enhanced morbidity, whereas those given 100 and 200 PFU of HPP-treated MNV-1 were comparable to uninfected controls. MNV-1 was detected, via real-time PCR, within the liver, spleen, and brain of all mice fed non-HPP-treated homogenate but was not detected in the tissues of mice fed HPP-treated homogenates or in uninfected control mice. Hepatocellular necrosis and lymphoid depletion in the spleen were observed in non-HPP-treated MNV-1 mice only. These results clearly show that HPP prevents MNV-1 infection in vivo and validates that viral inactivation by HPP in vitro is essentially equivalent to that in vivo. Further, the data suggest that HPP may be an effective food processing intervention for norovirus-contaminated shellfish and thus may decrease risk to both immunocompromised and immunocompetent individuals who consume shellfish., (Copyright ©, International Association for Food Protection)

Oocysts are the environmentally resistant life stage of Toxoplasma gondii. Humans can become infected by accidentally ingesting the oocysts in water or from contaminated produce. Severe disease can occur in immunocompromised individuals, and nonimmune pregnant women can infect their offspring. Chronic infection is associated with decreased mental functions, vision and hearing problems, and some mental disorders such as schizophrenia. High pressure processing (HPP) is a commercial method used to treat food to eliminate pathogens. Treatment of produce to eliminate viable T. gondii oocysts would provide a means to protect consumers. The present study was done to better define the effects of HPP on oocysts placed on raspberries. Raspberries were chosen because they are a known source of a related human intestinal parasite, Cyclospora cayetanensis. Raspberries were inoculated with 5 x 10(4) oocysts of the VEG strain of T. gondii for 20 hr prior to HPP. Individual raspberries were exposed to 500 MPa, 400 MPa, 340 MPa, 300 MPa, 270 MPa, 250 MPa, 200 MPA, 100 MPa, or no MPa treatment for 60 sec in a commercial HPP unit (1 MPa = 10 atm = 147 psi). Treatment of raspberries with 340 MPa for 60 sec was needed to render oocysts spot inoculated on the raspberries noninfectious for mice. Treatment of raspberries with 200 MPa or less for 60 sec was not effective in rendering oocysts noninfectious for mice.

The effectiveness of hydrostatic pressure processing (HPP) for inactivating viruses has been evaluated in only a limited number of studies, and most of the work has been performed with viruses freely suspended in distilled water. In this work, HPP inactivation of freely suspended and shellfish-associated bacteriophage T7 was studied. T7 was selected in hopes that it could serve as a model for animal virus behavior. Clams (Mercenaria mercenaria) and oysters (Crassostrea virginica) were homogeneously blended separately and inoculated with bacteriophage T7. The inoculated bivalve meat and the freely suspended virus samples were subjected to HPP under the following conditions: 2, 4, and 6 min at 241.3, 275.8, and 344.7 MPa pressure and temperatures of 29.4 to 35, 37.8 to 43.3, and 46.1 to 51.7 degrees C. Reductions of 7.8 log PFU (100% inactivation) were achieved for freely suspended T7 at 344.7 MPa for 2 min at 37.8 to 43.3 degrees C. At 46.1 to 51.7 degrees C, T7 associated with either clams or oysters was inactivated at nearly 100% (>4 log PFU) at all pressure levels and durations tested. These results indicate that T7 is readily inactivated by HPP under the proper conditions, may be made more susceptible to HPP by mixing with shellfish meat, and may serve as a viable model for the response of several animal viruses to HPP.

High hydrostatic pressure processing (HPP) has been shown to be an effective non-thermal means of inactivating microorganisms from various food products. Little information is available regarding the effects of HPP on metazoan parasites. Outbreaks of food-borne disease have been associated with importation of food contaminated with fecal material. Ascaris suum is used as a surrogate model metazoan parasite for the human roundworm, Ascaris lumbricoides, to study the effects of treatments on the inactivation of eggs in sludge. The present study was conducted to determine the effects of HPP on A. suum eggs. Unembryonated A. suum eggs were subjected to 138-552 megapascals (MPa) for 10-60s in a commercial HPP unit. Embryonation was induced after HPP treatments by incubating eggs in 0.01N sulfuric acid at room temperature. After 21 days, 100 eggs were examined per treatment using a light microscope and the percent of embryonated eggs was determined. Embryonation was induced in 38-76% eggs that were subjected to 138 and 270MPa. No embryonation was observed in eggs exposed to pressures of 241MPa or more for 60s or in eggs exposed to 276MPa for 10-30s. These results indicate that HPP treatment could be used to protect contaminated food items by inactivating A. suum eggs and may also have potential in reducing food-borne illness resulting from fecal contamination.

Murine norovirus (strain MNV-1), a propagable norovirus, was evaluated for susceptibility to high-pressure processing. Experiments with virus stocks in Dulbecco's modified Eagle medium demonstrated that at room temperature (20 degrees C) the virus was inactivated over a pressure range of 350 to 450 MPa, with a 5-min, 450-MPa treatment being sufficient to inactivate 6.85 log(10) PFU of MNV-1. The inactivation of MNV-1 was enhanced when pressure was applied at an initial temperature of 5 degrees C; a 5-min pressure treatment of 350 MPa at 30 degrees C inactivated 1.15 log(10) PFU of virus, while the same treatment at 5 degrees C resulted in a reduction of 5.56 log(10) PFU. Evaluation of virus inactivation as a function of treatment times ranging from 0 to 150 s and 0 to 900 s at 5 degrees C and 20 degrees C, respectively, indicated that a decreasing rate of inactivation with time was consistent with Weibull or log-logistic inactivation kinetics. The inactivation of MNV-1 directly within oyster tissues was demonstrated; a 5-min, 400-MPa treatment at 5 degrees C was sufficient to inactivate 4.05 log(10) PFU. This work is the first demonstration that norovirus can be inactivated by high pressure and suggests good prospects for inactivation of nonpropagable human norovirus strains in foods.

Four strains of Enterobacter sakazakii were inoculated into tryptic soy broth and reconstituted powdered infant formula and exposed to high-pressure processing. Pressures of 200, 400, and 600 MPa were used for each medium for 1 min. E. sakazakii was reduced by more than 6 log (strains A and B) in both media at 600 MPa. Strain B was significantly (P < or = 0.05) more pressure resistant than the other strains, with just more than a 3-log reduction at 600 MPa in both media. The reconstituted infant formula has a significant (P < or = 0.05) protective effect for certain strains and pressures (strain B at 400 MPa and strain D at 400 and 600 MPa). Differences in log reductions between media (milk and broth) were also observed for certain strains and specific pressures (strain B at 400 MPa and strain D at 400 and 600 MPa; P < or = 0.05). This research showed that E. sakazakii, when present in reconstituted powdered infant formula, can be submitted to high-pressure processing (600 MPa for 1 min) and achieve log reductions ranging from 3 to 6.84, depending on the strain.

Ingestion of Toxoplasma gondii tissue cysts can result in severe disease in immunocompromised individuals and pregnant women. Treatment of meat and meat products to eliminate viable T. gondii tissue cysts would provide a means to protect consumers. In this study, we examined the effects of high-pressure processing (HPP) on ground pork containing viable tissue cysts of the VEG strain of T. gondii. Ground pork containing tissue cysts was exposed to 400, 300, 200, 100, or 0 MPa treatment for 30, 60, or 90 sec in a commercial HPP unit. The HPP-treated ground pork was subjected to acid-pepsin digestion and bioassayed in mice. The results of the mouse bioassay revealed that none of the mice inoculated with tissue cysts exposed to 400 or 300 MPa became infected, whereas all mice inoculated with tissue cysts exposed to 200, 100, or 0 MPa became infected with T. gondii regardless of exposure time. Results indicate that HPP treatment of ground pork with 300 MPa of pressure will render tissue cysts of T. gondii nonviable and make pork safe for human consumption.

High-pressure processing (HPP) has been shown to be an effective means of eliminating bacteria and destructive enzymes from a variety of food products. HPP extends the shelf life of products while maintaining the sensory features of food and beverages. In this study, we examined the effects of HPP on the infectivity of Encephalitozoon cuniculi spores in vitro. Spores were exposed to between 140 and 550 MPa for 1 min in a commercial HPP unit. Following treatment, the spores were loaded onto cell culture flasks or were kept for examination by transmission electron microscopy. No effect was observed on the infectivity of spores treated with 140 MPa. Spores treated with between 200 and 275 MPa showed reduction in infectivity. Following treatment of 345 MPa or more, spores were unable to infect host cells. No morphologic changes were observed in pressure-treated spores with transmission electron microscopy.

Shellfish have been identified as a potential source of Cryptosporidium infection for humans. The inactivation of Cryptosporidium parvum and other pathogens in raw molluscan shellfish would provide increased food safety for normal and at-risk consumers. The present study identified the efficacy of a non-thermal alternative food-processing treatment, high hydrostatic pressure processing (HPP), on the viability of C. parvum oocysts in the Eastern oysters Crassostrea virginica. Oysters were artificially exposed to 2 x 10(7) oocysts of the Beltsville strain of C. parvum in seawater and subjected to HPP treatments. The effects of the treatments were evaluated by inoculation of the processed oyster tissues into neonatal mice. High-pressure processing of shucked Eastern oysters at all pressures tested (305, 370, 400, 480, and 550 MPa) was significantly effective (P<0.05) in reducing the numbers of positive mouse pups fed treated oyster tissues exposed to C. parvum oocysts. A dose of 550 MPa at 180 s (s) of holding time produced the maximum decrease in numbers of C. parvum positive mouse pups (93.3%). Measurement of tristimulus color values of HPP-treated raw oysters at extended processing times from 120 s to 360 s at 550 MPa showed a small increase in whiteness of oyster meat. This non-thermal processing treatment shows promise for commercial applications to improve safety of seafood and reduce public health risks from cryptosporidiosis.

Shellfish have been identified as a potential source of Cryptosporidium infection for humans. The inactivation of C. parvum and other pathogens in raw molluscan shellfish would provide increased food safety for normal and at-risk consumers. The present study examined the efficacy of two alternative food-processing treatments, e-beam irradiation and microwave energy, on the viability of C. parvum oocysts in Eastern Oysters (Crassostrea virginica), which were artificially infected with the Beltsville strain of C. parvum. The effects of the treatments were evaluated by oral feeding of the processed oyster tissues to neonatal mice. Significant reductions (P<0.05) in infectivity were observed for in-shell and shucked oysters treated with e-beam irradiation at doses of 1.0, 1.5, or 2 kGy vs. untreated controls. A dose of 2 kGy completely eliminated C. parvum infectivity and did not adversely affect the visual appearance of the oysters. Oyster tissue treated with microwave exposures of 1 s (43.2 degrees C), 2 s (54.0 degrees C), and 3 s (62.5 degrees C) showed a reduction in C. parvum mouse infectivity, but the effects were not significantly different (P>0.05) from controls. Microwave energy treatments at 2 and 3 s showed extensive changes in oyster meat texture and color. Thus, because of lack of efficacy and unacceptable tissue changes, microwave treatment of oysters is not considered a viable food-processing method.

High pressure processing (HPP) has been shown to be an effective non-thermal method of eliminating non-spore forming bacteria from a variety of food products. The shelf-life of the products is extended and the sensory features of the food are not or only minimally effected by HPP The present study examined the effects of HPP using a commercial scale unit on the viability of Toxoplasma gondii oocysts. Oocysts were exposed from 100 to 550 MPa for 1 min in the HPP unit and then HPP treated oocysts were orally fed to groups of mice. Oocysts treated with 550 MPa or less did not develop structural alterations when viewed with light microscopy. Oocysts treated with 550 MPa, 480 MPa, 400 Mpa, or 340 MPa were rendered noninfectious for mice. Mice fed oocysts treated with no or 100 to 270 MPa became infected and most developed acute toxoplasmosis and were killed or died 7 to 10 days after infection. These results suggest that HPP technology may be useful in the removal of T. gondii oocysts from food products.

Toxoplasma gondii has recently been recognized to be widely prevalent in the marine environment. It has previously been determined that Eastern oysters (Crassostrea virginica) can remove sporulated T. gondii oocysts from seawater and that oocysts retain their infectivity for mice. This study examined the long-term survival of T. gondii oocysts in oysters and examined how efficient oysters were at removing oocysts from seawater. Oysters in 76-L aquaria (15 oysters per aquarium) were exposed to 1 x 10(6) oocysts for 24 hr and examined at intervals up to 85 days postexposure (PE). Ninety percent (9 of 10) of these oysters were positive on day 1 PE using mouse bioassay. Tissue cysts were observed in 1 of 2 mice fed tissue from oysters exposed 21 days previously. Toxoplasma gondii antibodies were found in 2 of 3 mice fed oysters that had been exposed 85 days previously. In another study, groups of 10 oysters in 76-L aquaria were exposed to 1 x 10(5), 5 x 10(4), or 1 x 10(4) sporulated T. gondii oocysts for 24 hr and then processed for bioassay in mice. All oysters exposed to 1 x 10(5) oocysts were infected, and 60% of oysters exposed to 5 x 10(4) oocysts were positive when fed to mice. The studies with exposure to 1 x 10(4) oocysts were repeated twice, and 10 and 25% of oysters were positive when fed to mice. These studies indicate that T. gondii can survive for several months in oysters and that oysters can readily remove T. gondii oocysts from seawater. Infected filter feeders may serve as a source of T. gondii for marine mammals and possibly humans.

We have been collaborating since 1992 in studies on southern sea otters (Enhydra lutris nereis) as part of a program to define factors, which may be responsible for limiting the growth of the southern sea otter population. We previously demonstrated Toxoplasma gondii in sea otters. We postulated that cat feces containing oocysts could be entering the marine environment through storm run-off or through municipal sewage since cat feces are often disposed down toilets by cat owners. The present study examined the sporulation of T. gondii oocysts in seawater and the survival of sporulated oocysts in seawater. Unsporulated oocysts were placed in 15 ppt artificial seawater, 32 ppt artificial seawater or 2% sulfuric acid (positive control) at 24 C in an incubator. Samples were examined daily for 3 days and development monitored by counting 100 oocysts from each sample. From 75 to 80% of the oocysts were sporulated by 3 days post-inoculation under all treatment conditions. Groups of 2 mice were fed 10,000 oocysts each from each of the 3 treatment groups. All inoculated mice developed toxoplasmosis indicating that oocysts were capable of sporulating in seawater. Survival of sporulated oocysts was examined by placing sporulated T. gondii oocysts in 15 ppt seawater at room temperature 22-24 C (RT) or in a refrigerator kept at 4 C. Mice fed oocysts that had been stored at 4C or RT for 6 months became infected. These results indicate that T. gondii oocysts can sporulate and remain viable in seawater for several months.

The objective of this study was to determine the effect of normal microflora and Morganella morganii on histamine formation and olfactory acceptability in raw bluefish under controlled storage conditions. Fillets inoculated with and without M. morganii were stored at 5, 10, and 15 degrees C for 7 days. Microbial isolates from surface swabs were identified and screened for histidine decarboxylase activity. Olfactory acceptance was performed by an informal sensory panel. Histamine levels were quantified using high-performance liquid chromatography and fluorescence detection. While olfactory acceptance decreased, histamine concentration and bacterial counts increased. Storage temperature had a significant effect on histamine levels, bacterial counts, and olfactory acceptance of the bluefish. Inoculation with M. morganii had a positive significant effect on histamine formation for bluefish held at 10 and 15 degrees C (P < 0.0001). The results of the study will serve in supporting U.S. Food and Drug Administration (FDA) regulations regarding guidance and hazard levels of histamine in fresh bluefish.

Temperature abuse during raw oyster harvesting and storage may allow for the multiplication of natural spoilage flora as well as microbial pathogens, thus posing a potential health threat to susceptible consumers and compromising product quality. The objective of this study was to provide a scientific basis for determining whether different refrigeration and abuse temperatures for raw oysters would result in a spoiled product before it became unsafe. Raw shellstock oysters (Crassostrea virginica) purchased from a commercial Virginia processor were subjected to different temperature abuse conditions (7, 13, and 21 degrees C) over a 10-day storage period. Salinity, pH, halophilic plate count (HPC), total culturable Vibrio counts, and culturable Vibrio vulnificus counts were determined at each abuse condition. V. vulnificus isolates were confirmed by a specific enzyme-linked immunosorbent assay. Olfactory analysis was performed to determine consumer acceptability of the oysters at each abuse stage. The pH of the oysters decreased over time in each storage condition. The HPC increased 2 to 4 logs for all storage conditions, while olfactory acceptance decreased over time. V. vulnificus levels increased over time, reaching 10(5) to 10(6) CFU/g by day 6. The length of storage had a greater effect on the bacterial counts and olfactory acceptance of the oysters (P < 0.05) over time than did the storage temperature (P < 0.05).

The effects of storage temperatures and times on the microbiological quality and safety of hard-shelled quahog clams (Mercenaria mercenaria) were examined. Samples were stored at four different incubation temperatures (3.3, 7.2, 10.0, and 12.8 degrees C) for a period of 3 weeks, following their harvest from summer growing waters (> or = 27 degrees C) and winter waters (< or = 4 degrees C). Clams were analyzed for two naturally occurring pathogens, Vibrio parahaemolyticus and Vibrio vulnificus. During the summer, V. parahaemolyticus was isolated from 56% of the stored samples, with the highest concentration, 6,100/g, occurring on day 12 at 12.8 degrees C. Also, during the summer, V. vulnificus was isolated from 11% of the stored samples, with the highest concentration of 1,500/g occurring on day 15 at 7.2 degrees C. No Vibrio spp. were detected during the winter. During summer storage, aerobic mesophilic counts on plate count agar (PCA) containing 2% NaCl ranged from 10(4) to 10(8) CFU/g, and during storage of the winter samples, aerobic mesophilic PCA (with added NaCl) counts ranged from <100 to 10(4) CFU/g. Comparatively, summer storage mesophilic counts on PCA containing no added NaCl ranged from <100 to 10(5) CFU/g, and for the winter samples the range was <100 to 10(2) CFU/g. Coliform and fecal coliform counts ranged from <0.3 to 61.1/g and <0.3 to 24.4/g, respectively. There was no statistical correlation between the length of storage or the temperature of incubation and the presence of V. parahaemolyticus, V. vulnificus, coliforms, or fecal coliforms. However, storage time and incubation temperature affected the PCA counts (P < or = 0.05) in quahog clams.

Changes in histamine, putrescine, and cadaverine concentrations in bluefish filets (Pomatomus saltatrix) stored at 5, 10, and 15 degrees C were determined using high-performance liquid chromatography. An organoleptic assessment was conducted simultaneously with the biogenic amine analyses. The histamine levels found in fresh bluefish obtained from wholesale seafood distributors ranged between <1 ppm and 99 with an average of 39 ppm. Putrescine and cadaverine were not found in fresh bluefish. Fish fillets stored at each of the three temperatures developed histamine. The greatest accumulation of histamine was observed in fish stored at 15 degrees C, which developed histamine levels as high as 2,200 ppm. Putrescine levels increased at each temperature during storage. Cadaverine was present only in uninoculated bluefish stored at 15 degrees C. Histamine achieved higher levels in bluefish pieces inoculated with Morganella morganii, which demonstrates that bluefish support bacterial histamine formation. Histamine levels at each temperature exceeded the 50-ppm advisory level established by the Food and Drug Administration before 100% sensory rejection. Standard plate counts increased during storage of fish at all temperatures, but the correlation between histamine levels and standard plate count was not significant.

A microbial preparation derived from aquacultured channel catfish fillets (Ictalurus punctatus) was acidified with 0, 1, 2 and 4% (vol/vol) weak organic and held in an ice bath at 0 degree C to simulate the chilling process. Additionally, catfish fillets were sprayed under varying pressures at 15 degrees C with organic acids to evaluate the efficacy of concentrations of organic acids and spray pressures to ameliorate the microbiological quality. To determine plate counts, the dilution fluid was neutralized to pH 7.2 with 1.0 M NaOH. The aerobic plates counts of microorganisms in the chilling water were monitored over a 20-min interval. Aerobic plate counts were found on the channel catfish fillets before and after spray washing with organic acids. Plates were incubated at 35 degrees C for 48 h. The addition of organic acids tot he microbial preparation used in simulating the chilling process significantly (P < 0.05) reduced the number of bacterial surviving. The number of surviving bacteria in the chilled water decreased with increasing concentration and time of exposure to organic acids. Propionic acid had the most detrimental effect on organisms present in the microfloral preparation followed by acetic and lactic acids. Spray washing of catfish fillets with water did not significantly (P < 0.05) affect the microbial quality of fillets. However, catfish fillets sprayed with organic (lactic and propionic) acids significantly (P < 0.05) reduced the microbial counts by 10-fold. Lactic and propionic acids were not significantly (P > 0.05) different in influencing the aerobic counts of the catfish fillets.

Aquacultured rainbow trout (Oncorhynchus mykiss) and channel catfish (Ictalurus punctatus) fillets were inoculated with the psychrotrophic pathogens Listeria monocytogenes and Aeromonas hydrophila: cell populations were monitored during refrigerated storage at 2 to 4 degrees C. Fillets of both species were placed individually in sterile plastic bags and inoculated with cell suspensions (10(4.7) CFU/100 g of fish) of either A. hydrophila or L monocytogenes or of both A. hydrophila and L. monocytogenes, for a total of three treatments for each species of fish. Each inoculum and fillet were mixed to ensure uniform distribution and then stored at 2 to 4 degrees C. A. hydrophila, L. monocytogenes, and aerobic cell populations were determined on days 1, 3, 6, 8, 10, 13, and 15. Individually inoculated A. hydrophila and L. monocytogenes grew on catfish and trout fillets during the 15-day study. There was no inhibition of either pathogen by the natural flora on the fillets. Both psychrotrophic pathogens grew equally well in catfish and trout fillets inoculated with a combination of A. hydrophila and L. monocytogenes. In all three treatments, the counts of the psychrotrophic pathogens were lower than the aerobic plate counts. The growth of the psychrotrophic pathogens L. monocytogenes and/or A. hydrophila during refrigerated storage on aquacultured fish fillets could increase the food hazard risk, particularly where there is a possibility of cross-contamination with ready-to-eat food products.

The nature and number of indicator and pathogenic microbes in fish reared using recirculating and nonrecirculating water systems were compared. For each system, 20 samples of rainbow trout (Oncorhynchus mykiss), tilapia (Oreochromis spp.), hybrid striped bass (Morone saxatilis x M. chrysops), and pacu (Piaractus mesopotamicus) were randomly selected and gutted, and microbial analyses were performed using AOAC procedures. Five fish were subsampled and analyzed for indicative microbial quality with 3M Petrifilm (). The general microbial quality differed significantly (P < 0.05) among the production systems, except for total coliform counts. Rainbow trout cultured in recirculating and nonrecirculating water systems had lower counts for aerobes (2.00 to 3.11 log CFU/g) (p < 0.05), than other species, whereas trout reared in a recirculating water system had significantly lower psychrotrophic numbers (0.86 to 1.85 log CFU/g). Pacu had the highest fecal coliform counts (2.74 to 3.70 log CFU/g), whereas hybrid striped bass and rainbow trout grown in nonrecirculating systems had lower fecal coliform counts (0.00 to 1.39 log CFU/g). Rainbow trout grown in a nonrecirculating system had significantly higher Escherichia coli counts (0.00 to 2.11 log CFU/g). The human bacterial pathogens Listeria monocytogenes, Yersinia enterocolitica, Escherichia coli O157:H7 and Salmonella spp. were not isolated form the fish sampled. However, Clostridium botulinum botulinum was isolated from all the aquacultured fish sampled except pacu and tilapia grown in a recirculating aquaculture system. However, the counts were very low, ranging from 0.0 to 2.3 MPN/g.

Aquacultured channel catfish ( Ictalurus punctatus ) were evaluated for the presence of human pathogenic bacteria. Fresh catfish fillets procured from three catfish processors in the southeastern United States during the four annual seasons (e.g., summer, fall, winter, and fall) were screened for selected human pathogens. At each sampling time point, 20 freshly processed catfish fillets were randomly selected from each processor during each season. Five catfish fillets were randomly selected for aerobic plate counts and all 20 fillets were screened for five pathogenic bacteria viz. Campylobacter jejuni/coli , Escherichia coli O157:H7, Klebsiella pneumoniae subsp. pneumoniae , Plesiomonas shigelloides , and Vibrio cholerae . There was a significant difference ( P < 0.05) in the aerobic plate counts due to differences in unit processing operations and processing season. C. jejuni/coli , E. coli O157:H7 and K. pneumoniae subsp. pneumoniae were not isolated. Only P. shigelloides and V. cholerae were isolated during the warm weather.

Fresh aquacultured catfish fillets were obtained from three processors using different processing protocols in summer, autumn, winter, and spring and evaluated for microbial quality. Twenty freshly processed fillets were randomly selected and each fillet was placed in a sterile polyethylene bag. The fillets were transported on ice-pack overnight by air immediately after processing. Five fillets were randomly selected for microbial assays. Each fillet was weighed and an equal volume of sterile 0.1% peptone water at 0 to 1 degrees C was added aseptically. The fillet was massaged (or rinsed) for 120 s and the rinse was used to determine microbial quality. Aerobes (incubation at 35 degrees C for 48 h) and psychrotrophs (incubation at 20 degrees C for 96 h) were enumerated using 3M Petrifilm Aerobic Count plates. Escherichia coli (incubation at 35 degrees C for 24 to 48 h) and total coliforms (incubation at 35 degrees C for 24 to 48 h) were enumerated on 3M Petrifilm E. coli Count plates. Staphylococcus aureus counts were determined on Baird-Parker agar (incubation at 35 degrees C for 48 h). Significant differences (P < or = 0.05) in aerobic, psychrotrophic, total coliform, E. coli, and S. aureus counts due to temperature effects during production and variations in processing protocols were observed. E. coli and S. aureus counts were significantly different during the four seasons. E. coli and S. aureus counts were high during summer and low during winter weather. There was a significant difference (P < or = 0.05) in aerobic, psychrotrophic, and total coliform counts among the three processors during warm weather; however, these differences were significantly (P < or = 0.05) reduced in cold weather.

Fresh aquacultured catfish fillets were obtained from three processors using different processing protocols in summer, autumn, winter, and spring and evaluated for microbial quality. Twenty freshly processed fillets were randomly selected and each fillet was placed in a sterile polyethylene bag. The fillets were transported on ice-pack overnight by air immediately after processing. Five fillets were randomly selected for microbial assays. Each fillet was weighed and an equal volume of sterile 0.1 % peptone water at 0 to 1°C was added aseptically. The fillet was massaged (or rinsed) for 120 s and the rinse was used to determine microbial quality. Aerobes (incubation at 35°C for 48 h) and psychrotrophs (incubation at 20°C for 96 h) were enumerated using 3M Petrifilm™ Aerobic Count plates. Escherichia coli (incubation at 35°C for 24 to 48 h) and total coliforms (incubation at 35°C for 24 to 48 h) were enumerated on 3M Petrifilm™ E. coli Count plates. Staphylococcus aureus counts were determined on Baird-Parker agar (incubation at 35°C for 48 h). Significant differences ( P ≤ 0.05) in aerobic, psychrotrophic, total coliform, E. coli , and S. aureus counts due to temperature effects during production and variations in processing protocols were observed. E. coli and S. aureus counts were significantly different during the four seasons. E. coli and S. aureus counts were high during summer and low during winter weather. There was a significant difference ( P ≤ 0.05) in aerobic, psychrotrophic, and total coliform counts among the three processors during warm weather; however, these differences were significantly ( P ≤ 0.05) reduced in cold weather.

This study was conducted to determine the levels of Campylobacter jejuni , Campylobacter coli , Salmonella , Klebsiella pneumoniae , and Escherichia coli O157:H7 in fresh hand-picked blue crab ( Callinectes sapidus ) meat. An attempt was made to correlate these selected pathogens and general microbial quality to processors' sanitation practices and facility size. Hand-picked crabmeat samples from 12 blue-crab-processing facilities in the Chesapeake Bay region were collected and analyzed. Twenty samples from each of the different facilities were collected on different processing days. Facilities were chosen on the basis of production levels and Virginia Department of Health inspection scores as an indicator of sanitation practices (excellent, ≥94.5, acceptable, <94.5). All samples were tested between 16 h and 36 h after collection. Campylobacter jejuni was isolated from 36 (15%) of the 240 samples and Campylobacter coli was isolated from 14 (5.8%). Quantitative levels in all case were below limits of detection (<0.30 MPN/g). Klebsiella pneumoniae was isolated from a total of 51 (21%) samples. Counts ranged from less than 0.30 to 4.3 MPN/g. Aerobic plate counts ranged from 7.4 × 10 3 to 4.6 × 10 8 CFU/g with coliform counts ranging from <0.3 to 32.8 MPN/g. Fecal coliform levels were <0.3 to 2.26 MPN/g and Escherichia coli from <0.3 to 0.77 MPN/g. Salmonella spp. and Escherichia coli O157:H7 were not detected in any of the 240 samples analyzed. No significant differences ( P < 0.05) between size and inspection scores were observed for general microbial quality or the presence of Klebsiella pneumoniae or Campylobacter species.

There has been significant concern in recent times about the safety of molluscan shellfish for human consumption. Despite extensive efforts to assure a safe supply of molluscan shellfish, the number of cases of disease and death are still great enough to cause concern among the public. The number of cases of illness and death associated with the ingestion of shellfish falls in the lower end of the range of other similar microbial pathogen-related foodborne disease. Disease and deaths due to viruses and naturally occurring bacteria are now of greatest concern because they are the most often cited causative agents. The greatest risk of disease or death due to shellfish consumption is among the population with underlying health conditions who choose to consume raw shellfish. Control strategies to limit shellfish-borne disease should focus upon disease and death caused by viruses and naturally occurring bacteria among at-risk populations.

Fresh and 40-year-old pasteurized blue crab ( Callinectes sapidus ) meat was analyzed for proximate composition, mineral, heavy metal, and amino acid content and volatiles concentration and possible pesticide and herbicide residue levels. Both fresh and 40-year-old meat had similar proximate compositions. The 40-year-old crab meat contained high levels of iron, manganese, copper, and heavy metals compared to the fresh. Recovery of total amino acids was lower from the 40-year-old meat. Aspartic acid, glutamic acid, leucine, lysine, and arginine were the major protein amino acids of fresh and 40-year-old crab meat. The 40-year-old meat contained high concentrations of ethanol and trimethylamine compared to the fresh. No herbicide residues were detected in either of the products. Decomposition products of pesticide DDT were detected at very low levels only in the 40-year-old crab meat.

Tissues of Eastern ( Crassostrea virginica ) and Korean ( Crassostrea gigas ) oysters were analyzed for proximate composition; elemental, amino acid, and pesticide contents; and selected enzyme activities. Eastern oysters contained more nitrogen but a lower ash and lipid content. In general, the amino acid and elemental contents were higher in the Korean species. Neither the Eastern nor Korean species contained pesticides or PCBs in an amount greater than 0.1 ppm. Peroxidase activity was greater in Korean oysters while lipase activity was higher in the Eastern species. Lipoxygenase was not detected in either species.

The skin, flesh, and seed of the green variety of chayote ( Sechium edule ) were analyzed for relative phosphatase activities on phenyl phosphate, sodium phytate, fructose-1, 6-diphosphate, β-glycerol phosphate. glucose-1-phosphate, glucose-6-phosphate, and ATP. Relative rates of phosphatase activities were highest with ATP and lowest with glucose-1-phosphate. Activities of all phosphatases were highest in the skin and lowest in the seed.

Tissues of purple, green, and white varieties of eggplant, Solanum melongena , were analyzed for relative activity of phenyl phosphatase, fructose-1, 6-diphosphatase, glucose-1-phosphatase, glucose-6-phosphatase, and ATPase. Activities of all phosphatases were highest in the purple variety and lowest in the white. Relative rates of activity decreased in the order: ATPase, phenyl phosphatase, fructose-1, 6-diphosphatase, glucose-6-phosphatase, and glucose-1-phosphatase (only found in the purple variety).

Three different enzyme assays which have been used to determine peroxidase activity in food products were compared for their relative sensitivity to measure peroxidase in the blue crab ( Callinectes sapidus ). These included a modified Nagle and Haard, a Nickel and Cunningham, and a Lundquist and Josefsson procedure. The enzyme extracts were tested using a spectrophotometer to measure the ΔOD/min. The modified Nale and Haard procedure was the most sensitive method for detecting peroxidase activity in the blue crab. The assay, which is rapid, can be used with crude extracts and does not require enzyme purification.

Lipoxygenase activity in three cultivars (purple, green, and white) of eggplant, Solanum melongena, were compared. Activity was greatest in the purple and lowest in the white variety. In contrast to reports that NaCN did not inhibit eggplant lipoxygenase, in these studies cyanide completely inhibited the enzyme in all three varieties.