Species-specific impact of microplastics on coral physiology.

There is evidence that microplastic (MP) pollution can negatively influence coral health; however, mechanisms are unknown and most studies have used MP exposure concentrations that are considerably higher than current environmental conditions. Furthermore, whether MP exposure influences coral susceptibility to other stressors such as ocean warming is unknown. Our objective was to determine the physiology response of corals exposed to MP concentrations that have been observed in-situ at ambient and elevated temperature that replicates ocean warming. Here, two sets of short-term experiments were conducted at ambient and elevated temperature, exposing the corals Acroporasp. and Seriatopora hystrix to microspheres and microfibres. Throughout the experiments, gross photosynthesis and net respiration was quantified using a 4-chamber coral respirometer, and photosynthetic yields of photosystem II were measured using Pulse-Amplitude Modulated (PAM) fluorometry. Results indicate the effect of MP exposure is dependent on MP type, coral species, and temperature. MP fibres (but not spheres) reduced photosynthetic capability of Acropora sp., with a 41% decrease in photochemical efficiency at ambient temperature over 12 days. No additional stress response was observed at elevated temperature; photosynthetic performance significantly increased in Seriatopora hystrix exposed to MP spheres. These findings show that a disruption to coral photosynthetic ability can occur at MP concentrations that have been observed in the marine environment and that MP pollution impact on corals remains an important aspect for further research.

Effects of ultraviolet irradiation on chemical and sensory properties of goat milk.

Sensory and chemical consequences of treating goat milk using an UV fluid processor were assessed. Milk was exposed to UV for a cumulative exposure time of 18 s and targeted UV dose of 15.8 +/- 1.6 mJ/cm2. A triangle test revealed differences between the odor of raw milk and UV irradiated milk. Oxidation and hydrolytic rancidity was measured by thiobarbituric acid reactive substances and acid degree values (ADV). As UV dose increased, there was an increase in thiobarbituric acid reactive substance values and ADV of the milk samples. A separate set of samples were processed using the fluid processor but with no UV exposure to see if lipase activity and agitation from pumping contributed to the differences in odor. The ADV increased at the same rate as samples exposed to UV; however, sensory studies indicated that the increase of free fatty acids was not enough to cause detectable differences in the odor of milk. Solid phase microextraction and gas chromatography were utilized for the analysis of volatile compounds as a result of UV exposure. There was an increase in the concentration of pentanal, hexanal, and heptanal (relative to raw goat milk) after as little as 1.3 mJ/cm2 UV dose. Ultraviolet irradiation at the wavelength 254 nm produced changes in the sensory and chemical properties of fluid goat milk.

Efficacy of UV light for the reduction of Listeria monocytogenes in goat's milk.

Certain types of goat's cheeses are produced using unpasteurized milk, which increases the food safety concerns for these types of products. Popularity and consumption of goat's milk products have increased, and the niche market includes gourmet goat's cheeses. The U.S. Code of Federal Regulations and the Pasteurized Milk Ordinance both address the possibility for processing alternatives to heat treatment, and the use of UV light treatment may be a viable alternative that still ensures the safety of the product. Fresh goat's milk was inoculated with Listeria monocytogenes (L-2289) at 10(7) CFU/ml and exposed to UV light using the CiderSure 3500 apparatus (FPE Inc., Macedon, NY). Inoculated milk was exposed to a UV dose range between 0 and 20 mJ/cm2 to determine the optimal UV dose. A greater than 5-log reduction was achieved (P < 0.0001) when the milk received a cumulative UV dose of 15.8 +/- 1.6 mJ/cm2. The results of this study indicate that UV irradiation could be used for the reduction of L. monocytogenes in goat's milk.

Flavor threshold for acetaldehyde in milk, chocolate milk, and spring water using solid phase microextraction gas chromatography for quantification.

The detection threshold of acetaldehyde was determined on whole, lowfat, and nonfat milks, chocolate-flavored milk, and spring water. Knowledge of the acetaldehyde threshold is important because acetaldehyde forms in milk during storage as a result of light oxidation. It is also a degradation product of poly(ethylene terephthalate) during melt processing, a relatively new packaging choice for milk and water. There was no significant difference in the acetaldehyde threshold in milk of various fat contents, with thresholds ranging from 3939 to 4040 ppb. Chocolate-flavored milk and spring water showed thresholds of 10048 and 167 ppb, respectively, which compares favorably with previous studies. Solid phase microextraction (SPME) was verified as an effective method for the recovery of acetaldehyde in all media with detection levels as low as 200 and 20 ppb in milk and water, respectively, when using a polydimethyl siloxane/Carboxen SPME fiber in static headspace at 45 degrees C for 15 min.

Levels of Vibrio vulnificus and organoleptic quality of raw shellstock oysters (Crassostrea virginica) maintained at different storage temperatures.

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).

Nonproteolytic Clostridium botulinum toxigenesis in cooked turkey stored under modified atmospheres.

The ability of nonproteolytic Clostridium botulinum type B spores to grow and produce toxin in cooked, uncured turkey packaged under modified atmospheres was investigated at refrigeration and mild to moderate abuse temperatures. Cook-in-bag turkey breast was carved into small chunks, surface-inoculated with a mixture of nonproteolytic C. botulinum type B spores, packaged in O2-impermeable bags under two modified atmospheres (100% N2 and 30% CO2:70% N2), and stored at 4, 10, and 15 degrees C. Samples were analyzed for botulinal toxin and indigenous microorganisms, as well as subjected to sensory evaluation, on days 0, 7, 14, 28, 42, and 60. Given sufficient incubation time, nonproteolytic C. botulinum type B grew and produced toxin in all temperature and modified atmosphere treatment combinations. At moderate temperature abuse (15 degrees C), toxin was detected by day 7, independent of packaging atmosphere. At mild temperature abuse (10 degrees C), toxin was detected by day 14, also independent of packaging atmosphere. At refrigeration temperature (4 degrees C), toxin was detected by day 14 in product packaged under 100% N2 and by day 28 in product packaged under 30% CO2:70% N2. Reduced storage temperature significantly delayed toxin production and extended the period of sensory acceptability of cooked turkey, but even strict refrigeration did not prevent growth and toxigenesis by nonproteolytic C. botulinum. At all three storage temperatures, toxin detection preceded or coincided with development of sensory characteristics of spoilage, demonstrating the potential for consumption of toxic product when spoilage-signaling sensory cues are absent.

Efficacy of ultraviolet light for reducing Escherichia coli O157:H7 in unpasteurized apple cider.

This study examined the efficacy of UV light for reducing Escherichia coli O157:H7 in unpasteurized cider. Cider containing a mixture of acid-resistant E. coli O157:H7 (6.3 log CFU/ml) was treated using a thin-film UV disinfection unit at 254 nm. Dosages ranged from 9,402 to 61,005 microW-s/cm2. Treatment significantly reduced E. coli O157:H7 (P < or = 0.0001). Mean reduction for all treated samples was 3.81 log CFU/ml. Reduction was also affected by the level of background microflora in cider. Results indicate that UV light is effective for reducing this pathogen in cider. However, with the dosages used in this experiment, additional reduction measures are necessary to achieve the required 5-log reduction.

Growth and histamine formation of Morganella morganii in determining the safety and quality of inoculated and uninoculated bluefish (Pomatomus saltatrix).

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.

Effect of hydrogen peroxide and other protease inhibitors on Cryptosporidium parvum excystation and in vitro development.

This study was undertaken to observe the effects of hydrogen peroxide on Cryptosporidium parvum oocysts with respect to protease activity in comparison to known protease inhibitors. In assessing the possible mechanisms of action of hydrogen peroxide, treatment effectiveness was analyzed using 3 assays and the potential roles of proteases and cations were considered. Treatment of C. parvum oocysts with hydrogen peroxide inhibited protease activity up to 50% compared with untreated controls. Treatment of oocysts with chemicals that affect sulfhydryls, including N-ethylmaleimide and dithiolthreitol, inhibited protease activity by >90%. Treatment of oocysts with these chemicals, along with the protease inhibitors, phenylmethylsulfonyl fluoride (PMSF), ethylenediamine-tetraacetic acid, and cystatin, inhibited protease activity as well as in vitro excystation and infection in a cell culture assay. Several mechanisms may result in the successful inhibition of infection and excystation by hydrogen peroxide treatment, including: oxidation of oocyst wall proteins or lipids, chelating of cations necessary for infection, or hydroxyl radical-induced DNA damage to sporozoites, or both.

Comparison of sampling techniques for detection of Arcobacter butzleri from chickens.

Arcobacter butzleri is a causative agent of human enteritis that has been recently differentiated from the genus Campylobacter. Previous work suggests that its transmission to humans is likely through a foodborne route with a substantial tendency to be located on poultry carcasses. For reducing the incidence of this pathogen on commercial poultry, improved protocols are needed to sample and identify A. butzleri from infected birds prior to slaughter. The purpose of this study was to compare sampling methods for this emerging pathogen from chickens that were artificially inoculated per os with A. butzleri. We tested three sampling techniques commonly used to determine the microbiological quality of poultry: cloacal swabs, fecal samples, and environmental surface (drag) swabs collected when birds were 3, 5, or 7 wk old. These samples were cultured in Johnson-Murano enrichment broth and analyzed by PCR. Results indicate that environmental surface swabs yielded the highest recovery percentage. A detection rate of 75 to 100% was observed for each sampling period (age of chicken). Additionally, A. butzleri could not be isolated from the intestinal tract (jejunum, ileum, cecum, colorectum) of inoculated birds.

Enhanced recovery of injured Escherichia coli by compounds that degrade hydrogen peroxide or block its formation.

Escherichia coli LSUFS was injured either by freezing at -10 degrees C or by heating at 57 degrees C for 12 min. Surviving cells were recovered on nonselective tryptone-glucose extract agar and selective violet red bile agar supplemented with compounds that degrade hydrogen peroxide or block its formation. Various concentrations of the following compounds were tested: sodium pyruvate, 3,3'-thiodipropionic acid, catalase, ascorbic acid, potassium permanganate, sodium thioglycolate, dimethylsulfoxide, ethoxyquin, n-propyl gallate, alpha-tocopherol sodium metabisulfite, and ferrous sulfate. Sodium pyruvate and 3,3'-thiodipropionic acid, when added to either medium, significantly (P greater than 0.01) increased recovery of injured cells. More than 90% of the heat-injured cells and 40 to 90% of the freeze-injured cells failed to grow on unsupplemented tryptone-glucose extract agar. Supplementation of violet red bile agar increased recovery, but the counts remained considerably lower than the tryptone-glucose extract agar counts. The repair detection procedure of Speck et al. (M. Speck, B. Ray, R. Read, Jr., Appl. Microbiol. 29:549-550, 1975) was greatly improved by the addition of pyruvate or 3,3'-thiodipropionic acid. However, when this improved repair detection procedure was applied to foods, pyruvate-supplemented media showed some false-positives. We therefore recommend that 3,3'-thiodipropionic acid be used to supplement media in the repair detection procedure.

Incidence of Vibrio parahaemolyticus in and the Microbiological Quality of Seafood in North Carolina 1.

Microbiological analyses of 716 seafood samples over a 3-year period revealed that the microbiological quality of fresh seafood in North Carolina was generally acceptable. The mean aerobic counts (APC) and fecal coliform counts were low as was the occurrence of enteric pathogens, except for Vibrio parahaemolyticus and coagulase-positive staphylococci. Salmonella species were isolated from three samples, but the fecal coliform counts of these samples far exceeded the shellfish standard of the United States Food and Drug Administration. Coagulase-positive staphylococci were isolated in low numbers from nearly all the different types of seafoods; unpasteurized crabmeat and head-peeled shrimp samples showed the highest counts (10% of these samples had numbers which exceeded 100/g). V. parahaemolyticus occurred frequently in fresh seafood (overall 46% of the samples were positive) and its numbers showed a definite seasonal variation. No positive statistical correlation was found between the numbers of V. parahaemolyticus and the bacteriological indices, such as coliforms, fecal coliforms, enterococci and APC. Processing practices were found to influence the occurrence of V. parahaemolvticus in seafood; for example, improperly cleaned flumes were found to be a reservoir for V. parahaemolyticus in mechanical scallop processing plants. Also 'picking' waste 'containers' were found to be sources of V. parahaemolyticus in crab processing plants. Some processing practices such as heat shocking of oysters to facilitate opening were found to reduce the numbers of V. parahaemolyticus . Fifty V. parahaemolyticus isolates from different seafoods were tested for their Kanagawa reaction and all were found to be negative.

Method for the detection of injured Vibrio parahaemolyticus in seafoods.

The sensitivity of Vibrio parahaemolyticus cells to refrigeration and frozen storage and the development of a method for detecting injured and uninjured V. parahaemolyticus cells were studied. Cell suspensions in different kinds of seafood homogenates were either regrigerated (4 degrees C) or frozen (-20 degrees C), stored, and examined for cell survival during storage. V. parahaemolyticus cells were sensitive to both storage temperatures. Many cells died, and many survivors were sublethally injured. In general, refrigeration storage appeared to be more injurious than frozen storage. The initial recovery of the sublethally injured cells was highest in a nutritionally rich, nonselective liquid medium such as Trypticase soy broth, whereas maximum cell multiplication was observed in Trypticase soy broth containing 3% NaCl. The sublethally injured V. parahaemolyticus cells demonstrated sensitivity to the selective enrichment medium, glucose salt teepol broth. From these findings, a new method (designated as the "repair-detection" method) was developed for the isolation and enumeration of V. parahaemolyticus. Comparative studies between the recommended and the repair-detection methods showed that injured V. parahaemolyticus cells were present in commercial seafoods and that the repair-detection method was definitely more effective for the detection of total numbers of V. parahaemolyticus cells.

Repair detection procedure for enumeration of fecal coliforms and enterococci from seafoods and marine environments.

The repair detection procedure of Speck et al. (Appl. Microbiol. 29:549-550, 1975) was adapted for the enumeration of coliforms, fecal coliforms, and enterococci in seafood and environmental samples. Samples were pour plated with Trypticase soy agar, followed by a 1- to 2-h incubation to effect repair; the plates were then overlaid with the selective medium and incubated. Violet red bile agar and an incubation temperature of 45 degrees C were used as the selective conditions for fecal coliforms, and KF streptococcal agar was used for the enumeration of enterococci. The method was more efficient than the standard most-probable-number method for fecal coliform enumeration and also allowed enumeration of the injured cells, which might have remained undetected when selective medium in the most-probable-number method was used. The repair detection method effectively recovered the injured portion of the population of enterococci capable of growing on KF streptococcal agar. The repair enumeration method was not suitable for coliforms in marine samples because associative marine bacteria mimicked coliforms in violet red bile agar plates incubated at 35 degrees C. The marine bacteria did not grow at 45 degrees C and therefore did not interfere with fecal coliform enumeration.

Adherence as a method of differentiating virulent and avirulent strains of Vibrio parahaemolyticus.

Usually only Kanagawa-positive strains of Vibrio parahaemolyticus are considered virulent; yet, a significant portion of V. parahaemolyticus food poisonings appear to be caused by Kanagawa-negative strains. Therefore, additional and more accurate measurements of a strain's food-poisoning potential are needed. Adherence of V. parahaemolyticus to human fetal intestinal (HFI) cells in vitro seems to offer this information. All strains of V. parahaemolyticus adhered to the HFI cells, but the degree of adherence was related to a number of factors. These included the age of the culture, the strain's Kanagawa reaction and source, the length of time the bacteria were exposed to the HFI cells, and the composition of the growth medium. Cells harvested during the late log phase of growth adhered more intensely than those harvested from the late stationary phase. Virulent strains, i.e., those involved in food poisoning, were observed to have a high adherence ability regardless of their Kanagawa reaction, whereas Kanagawa-negative strains isolated from seafood exhibited weak adherence intensities. Kanagawa-positive strains isolated from seafood adhered strongly to the HFI cells. The difference between the virulent and avirulent strains was quantitative in nature, and the greatest differences in adherence intensities were observed after short (10 to 15 min) exposure times. The presence of ferric iron in the growth medium was found to increase the adherence intensities of the virulent strains.

Use of a Modified Gompertz Equation to Model Nonlinear Survival Curves for Listeria monocytogenes Scott A.

The heat resistance of Listeria monocytogenes was determined in 0.1 M KH2PO4 buffer at three temperatures (50, 55, and 60°C), three pH levels (5, 6, and 7), and three NaCl concentrations (0, 2, and 4%). Survival curves were fit using nonlinear regression with a modified Gompertz equation. The Gompertz equation is capable of fitting survival curves which are linear, those which display an initial lag region followed by a linear region, and those which are sigmoidal. Parameter estimates were used to describe the lag region, death rate, and the tailing region of a survival curve. These estimates were also used to predict single and interactive effects of temperature, pH, and percentage of NaCl on the log surviving fraction (LSF) of bacteria. Interactions among these variables significantly (P < .05) affected the LSF. Generally, increased pH or NaCl concentration lead to an increased (P < .05) LSF, where as increased time or temperature lead to a decreased (P < .05) LSF. All multiple factor interactions significantly (P < .05) affected the LSF. These interactions differed depending on the heating medium and the region of the survival curve. The correlation of observed LSF and predicted LSF (R2 = .89) indicated that the Gompertz equation was in close agreement with the observations. This study demonstrated that the Gompertz equation and nonlinear regression can be used as an effective means to predict survival curve shape and response to heat of L. monocytogenes in many different environmental conditions.

The Effect of Sublethal Heat Shock and Growth Atmosphere on the Heat Resistance of Listeria monocytogenes Scott A.

Log phase cells of Listeria monocytogenes Scott A were heat shocked in Trypticase Soy + 0.6% yeast extract (TSYE) broth at 48°C for 10 min, followed by heating at 55°C for up to 50 min. Heat resistance was determined using nonselective (TSYE) and selective (McBride Listeria ) enumeration media which were incubated under aerobic and anaerobic environments. D55°C-values for heat shocked cells were 2.1-fold higher than nonheat shocked cells (18.7 min vs. 8.89 min) when cells were enumerated on TSYE agar aerobically and 2.2-fold higher (26.4 min vs. 12.0 min) for cells enumerated anaerobically on TSYE agar. When cells were enumerated aerobically on McBride Listeria (ML) agar, D55°C-values for heat shocked cells were 1.4-fold higher than nonheat shocked cells (9.55 min vs. 6.69 min). No growth was observed on ML agar anaerobically. The physiological condition of the microorganism, the enumeration medium, and the growth environment greatly affected the heat resistance of logphase cells of Listeria monocytogenes Scott A.

Effectiveness of poly(ethylene terephthalate) and high-density polyethylene in protection of milk flavor.

The development of certain off-flavors in whole milk (3.25% milk fat) as related to packaging material [glass, high-density polyethylene (HDPE), amber poly(ethylene terephthalate) (PETE), clear PETE, and clear PETE-UV] were evaluated after exposure to fluorescent light (1100 to 1300 lx) for 18 d at 4 degrees C. Control samples packaged and stored under identical conditions were wrapped in foil to prevent light exposure. Selected flavor compounds in milk were measured analytically on d 0, 7, 14, and 18 of storage, while intensities of "oxidation," "acetaldehyde," and "lacks freshness" off-flavors were determined by sensory analysis at the same intervals. In light-exposed samples, oxidation off-flavor was significantly lower when packaged in amber PETE versus other containers. Milk packaged in HDPE containers showed a significantly higher level of oxidation off-flavor than milk packaged in PETE-UV containers but not higher than clear PETE or glass containers. No significant difference in acetaldehyde off-flavor was found between package material treatments (exposed or protected). Acetaldehyde concentration never exceeded flavor threshold levels, regardless of packaging material. Amber and PETE-UV materials proved to be a competitive packaging choice for milk in preserving fresh milk flavor.

Use of a Modified Gompertz Equation to Predict the Effects of Temperature, pH, and NaCl on the Inactivation of Listeria monocytogenes Scott A Heated in Infant Formula.

The heat resistance of Listeria monocytogenes was determined in infant formula for all possible combinations of temperature (50, 55, and 60°C), pH level (5, 6, and 7), and NaCl concentration (0, 2, and 4%). Survival curves were fit using nonlinear regression with a Gompertz equation. The Gompertz equation was flexible enough to fit the three most commonly observed survival curves: linear curves, those with an initial lag region followed by a linear region, and sigmoidal shaped. Parameter estimates obtained by the method of nonlinear least squares were used to describe the effect(s) of different heating treatments on the lag region, death rate, and tailing region of survival curves. These estimates were further used to predict single and interactive effects of temperature, pH, and percentage of NaCl on the log of the surviving fraction (LSF) of bacteria. Interactions among these variables significantly (P ≤ .05) affected the LSF. Generally, increased pH or NaCl concentration lead to an increased LSF, whereas increased time or temperature lead to a decreased LSF. All multiple-factor interactions significantly (P ≤ .05) affected the LSF. The correlation of observed LSF versus predicted LSF (R2 = .92) indicated that the estimated Gompertz equation was in close agreement with the observation. This study demonstrated that the Gompertz equation and nonlinear regression can be used as an effective means to predict survival curve shape and response to heat of L. monocytogenes under many different environmental conditions.

Probiotic culture survival and implications in fermented frozen yogurt characteristics.

Low-fat ice cream mix was fermented with probiotic-supplemented and traditional starter culture systems and evaluated for culture survival, composition, and sensory characteristics of frozen product. Fermentations were stopped when the titratable acidity reached 0.15% greater than the initial titratable acidity (end point 1) or when the pH reached 5.6 (end point 2). Mix was frozen and stored for 11 wk at -20 degrees C. The traditional yogurt culture system contained the strains Streptococcus salivarius ssp. thermophilus and Lactobacillus delbrueckii ssp. bulgaricus. The probiotic-supplemented system contained the traditional cultures as well as Bifidobacterium longum and Lactobacillus acidophilus. We compared recovery of Bifodobacterium by three methods, a repair-detection system with roll-tubes and plates on modified bifid glucose medium and plates with maltose + galactose reinforced clostridial medium. Culture bacteria in both systems did not decrease in the yogurt during frozen storage. The roll-tube method with modified bifid glucose agar and repair detection system provided at least one-half log10 cfu/ml higher recovery of B. longum compared with recoveries using modified bifid glucose agar or maltose + galactose reinforced clostridial agar on petri plates. No change in concentrations of lactose or protein for products fermented with either culture system occurred during storage. Acid flavor was more intense when product was fermented to pH 5.6, but yogurt flavor was not intensified. The presence of probiotic bacteria in the supplemented system seemed to cause no differences in protein and lactose concentration and sensory characteristics.