Microbial Indicators and Possible Focal Points of Contamination during Production and Processing of Catfish.

The catfish industry is important to the United States economy. The present study determined the levels of microbial indicators and the prevalence of Listeria spp. and Listeria monocytogenes at catfish farms and catfish processing plants. Live fish, water, and sediment samples were analyzed in farms. Fish skin, fillets, chiller water, and environmental surfaces were assessed at the processing plants both during operation and after sanitation. Live fish had 2% prevalence of Listeria monocytogenes, while sediment and water were negative for Listeria. Live fish skin counts averaged 4.2, 1.9, and 1.3 log CFU/cm2 aerobic (APC), total coliform (TCC) and generic Escherichia coli counts, respectively. Water and sediment samples averaged 4.8 and 5.8 log CFU/g APC, 1.9 and 2.3 log CFU/g TCC, and 1.0 and 1.6 log CFU/g generic E. coli counts, respectively. During operation, Listeria prevalence was higher in fillets before (57%) and after (97%) chilling than on fish skin (10%). Process chiller water had higher (p ≤ 0.05) APC, TCC, and Listeria prevalence than clean chiller water. After sanitation, most sampling points in which Listeria spp. were present had high levels of APC (>2.4 log CFU/100 cm2). APC combined with Listeria spp. could be a good approach to understand microbial contamination in catfish plants.

A New Approach for Quantifying Purpurogallin in Brewed Beverages Using LC-MS in Combination with Solid Phase Extraction.

Purpurogallin (PPG) is a phenolic compound known for its high antioxidant properties in plant-based food materials. However, there is no easy and reliable method for direct determination of PPG in brewed beverages owing to its hydrophobicity, which makes it hard to separate from the background hydrophobic components. Therefore, a method employing solid-phase extraction (SPE) and liquid chromatography-mass spectrometry (LC-MS) was developed for detection and quantification of PPG in brewed beverages, and PPG content was quantified in commercial coffee, cocoa, and tea samples. The limits of detection and quantification were 71.8 and 155.6 ng/g dry weight (dw), respectively. The recovery with SPE was 26.6%. When combined with acetonitrile extraction (ANE), the recovery was 6.8%, higher than 2.6% with water extraction (WTE). Test tube extractions were better than moka pot brewing (MPB) for PPG quantification. Total PPG content of ground coffees prepared by ANE, WTE, and MPB ranged between 635 and 770, 455 and 630, and 85 and 135 ng/g dw, respectively. PPG was detected in two English breakfast tea samples (335−360 ng/g dw) using WTE, but not in cocoa samples. ANE showed higher (p < 0.05) PPG levels, but WTE (r = 0.55, p < 0.01) correlated better with MPB than ANE (r = 0.43, p < 0.01). The result indicated that WTE is the best method to determine PPG in brewed beverages. This work demonstrated that PPG was significant in brewed coffee, and our pioneer study in developing the method for beverage sample preparation and LC-MS analysis has made possible industrial applications and provided new perspectives for future research.

Inactivation Mechanism of Aspergillus flavus Conidia by High Hydrostatic Pressure.

This study investigated the inactivation mechanism of Aspergillus flavus conidia by high hydrostatic pressure (HHP). Activity counts, scanning electron microscopic (SEM) analysis, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were used to study the effects of the HHP treatment on the morphology and protein composition of A. flavus spores. The results showed that that a 3-min-lasting 600 MPa treatment could completely abolish 107 colony-forming units/mL of live fungi. Furthermore, we also observed that lower spore viability corresponded to a higher Propidium Iodide absorption rate. The SEM images revealed that HHP disrupted the spore morphology and resulted in pore formation that led to the release of intracellular molecules, such as nucleic acids and proteins. The nucleic acid and protein concentration in the spore suspension increased in parallel with the increasing treatment pressure. The SDS-PAGE analysis showed that there were differences in the protein bands between the HHP-treated and untreated A. flavus spores, as the HHP treatment caused partial protein degradation and extracellular release. Therefore, the results of this study proved that high pressure could induce a morphological disruption in the internal and external cellular structures and degrade intracellular and extracellular proteins leading to an inactive state in A. flavus.

Influence of Microbial Transglutaminase on Physicochemical and Cross-Linking Characteristics of Individual Caseins.

The effects of microbial transglutaminase (MTGase) cross-linking on the physicochemical characteristics of individual caseins were investigated. MTGase was used to modify three major individual caseins, namely, κ-casein (κ-CN), αS-casein (αS-CN) and ß-casein (ß-CN). The SDS-PAGE analysis revealed that MTGase-induced cross-linking occurred during the reaction and that some components with high molecular weights (>130 kDa) were formed from the individual proteins κ-CN, αS-CN and ß-CN. Scanning electron microscopy (SEM) and particle size analysis respectively demonstrated that the κ-CN, αS-CN and ß-CN particle diameters and protein microstructures were larger and polymerized after MTGase cross-linking. The polymerized κ-CN (~749.9 nm) was smaller than that of ß-CN (~7909.3 nm) and αS-CN (~7909.3 nm). The enzyme kinetics results showed KM values of 3.04 × 10-6, 2.37 × 10-4 and 8.90 × 10-3 M for κ-CN, αS-CN and ß-CN, respectively, and, furthermore, kcat values of 5.17 × 10-4, 1.92 × 10-3 and 4.76 × 10-2 1/s, for κ-CN, αS-CN and ß-CN, respectively. Our results revealed that the cross-linking of ß-CN catalyzed by MTGase was faster than that of αS-CN or κ-CN. Overall, the polymers that formed in the individual caseins in the presence of MTGase presented a higher molecular weight and larger particles.

Green synthesis, characterization and evaluation of catalytic and antibacterial activities of chitosan, glycol chitosan and poly(γ-glutamic acid) capped gold nanoparticles.

Gold nanoparticles capped with chitosan (CH-NGs), glycol chitosan (GC-NGs) and poly(γ-glutamic acid) (PA-NGs) were synthesized separately, characterized and evaluated for catalytic and antibacterial activities. Surface Plasmon resonance peak at 520-530 nm confirmed the formation of NGs, while FTIR spectra revealed the involvement of hydroxyl, amine and amide groups in biopolymers on NGs formation and coating. Particle size, zeta potential and surface coating were respectively 21.7 nm, +50.2 mV and 20% for CH-NGs, 5.6 nm, +46.5 mV and 43.5% for GC-NGs and 7.4 nm, -37.3 mV and 34.5% for PA-NGs. Compared to citrate-capped NGs (CT-NGs), biopolymer-capped NGs exhibited high catalytic activity in a 4-nitrophenol reduction model with the pseudo first-order catalytic rate for PA-NGs being 4-6 fold higher than CH-NGs and GC-NGs. No significant antibacterial effect was shown for CT-NGs. However, PA-NGs was superior to gentamycin in inhibiting Salmonella enterica and Escherichia coli-O157:H7, while CH-NGs and GC-NGs showed the highest antibacterial effect against Listeria monocytogenes, followed by Salmonella enterica, Escherichia coli-O157:H7, methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus aureus. TEM images showed that GC-NGs were attached on MRSA surface to alter cell permeability, block nutrient flow and disrupt cell membrane, whereas PA-NGs penetrated into Salmonella enterica to generate cavities, plasmolysis and disintegration.

Effects of high-pressure processing and thermal pasteurization on quality and microbiological safety of jabuticaba (Myrciaria cauliflora) juice during cold storage.

The aims of this study were to investigate the effects of high-pressure processing (HPP) on jabuticaba juice characteristics including microbial levels, phenolic compounds, antioxidant activity, and physicochemical properties during 28 days of storage at 4 °C and to perform a sensory evaluation. Juice samples were pressurized at 200, 400, or 600 MPa for 5 min. During thermal processing, juice was treated in a water bath at 90 °C for 60 s. Elevated aerobic plate counts, coliforms, psychrotrophs and yeasts/molds, were not detected in the HPP-400, HPP-600, or thermal-processed (TP) juices and further cold storage showed at least a shelf life of 28 days at 7 °C. All HPP-treated juice had significantly higher antioxidant capacities, higher total phenolic, flavonoid, and monomeric anthocyanin content, and lower browning degrees, compared with the TP. The soluble solid content, titratable acidity and pH were not significantly different in the HPP-400, HPP-600, and TP after 28 days. The ΔE values were significantly increased in all juice samples. Sensory analysis indicated that the HPP-treated juices had higher acceptance and lower bitter perception. In conclusion, HPP treatments above 400 MPa were effective in ensuring microbiological safety, maintaining the overall quality parameters, extending the shelf life, and achieving consumer acceptance.

Extraction of bioactive ingredients from fruiting bodies of Antrodia cinnamomea assisted by high hydrostatic pressure.

The aim of this study was to use high hydrostatic pressure treatment to enhance the extraction efficiency of the active components from the fruiting bodies of Antrodia cinnamomea, and compare with those obtained by shake and ultrasonic extraction methods. The conditions of high pressure extraction (HPE) at 600 MPa, a liquid/solid ratio of 40:1, and 3 min of treatment yielded triterpenoids and adenosine concentrations of 410.41 mg/100 mL and 0.47 mg/100 mL, respectively, which did not differ significantly from those with the two other treatments-shake extraction at 180 rpm for 8 h and ultrasonic extraction at 50 Hz for 60 min. The HPE extracts significantly attenuated reactive oxygen species, nitric oxide and prostaglandin E2 production in lipopolysaccharide-stimulated RAW 264.7 cells than shake extracts did. SEM micrographs revealed that high-pressure caused physical morphological damage to the mycelium of fruiting bodies, such as distortion and disruption of mycelial cells, and increased the mass-transfer effectiveness of the solvent and solute. HPE can be employed as an efficient extraction technique for production of bioactive ingredients that might have a potential application in food and related industries.

Effects of high pressure extraction on the extraction yield, phenolic compounds, antioxidant and anti-tyrosinase activity of Djulis hull.

The hulls of Djulis (Chenopodium formosanum) are a type of agricultural waste. Using 70% ethanol as the extraction solvent, this study compared the extraction yields of high-pressure-assisted extraction (HPE) and conventional oscillation extraction (CE) for Djulis hulls (DH). The total phenolic and flavonoid contents, and antioxidant, anti-inflammatory and anti-tyrosinase activities were also compared. Our findings indicated that 600 MPa/5 min of HPE resulted in higher total phenolic (567-642 mg GAE/g) and flavonoid (47.2-57.2 mg QU/g) concentrations; gallic acid (44.5-53.2 µg/g) and rutin (26.8-34.2 µg/g) were the main phenolic and flavonoid compounds. When the extraction pressure was greater than 450 MPa, HPE extracts showed stronger antioxidant capacity and anti-tyrosinase activity than CE extracts. In a LPS-induced RAW 264.7 cell model of inflammation, HPE extracts had significant inhibitory effects on the cumulative concentrations of nitric oxide and prostaglandin E2. These results indicate that HPE had a better extraction yield, and required a shorter time for the extraction of functional ingredients from DH. Hence, DH could be a potential source for natural antioxidants for the food and biotechnology industries.

Antimicrobial Effects and Mechanisms of Ethanol Extracts of Psoralea corylifolia Seeds Against Listeria monocytogenes and Methicillin-Resistant Staphylococcus aureus.

Psoralea corylifolia seeds contain many bioactive compounds commonly used in traditional Chinese medicine. In this study, the antibacterial activity and possible mechanism of P. corylifolia seed ethanol extract (PCEE) against foodborne pathogens were investigated. Both methicillin-resistant Staphylococcus aureus (MRSA) and Listeria monocytogenes had similar minimum inhibitory concentrations and minimum bactericidal concentrations of PCEE at 50 and 100 µg/mL, respectively. Furthermore, elevated OD260, protein concentration, and electric conductivity indicated irreversible damage to the cytoplasmic membranes of PCEE-treated cells. Indeed, the treated cells displayed disrupted membranes, incomplete and deformed shapes, and rupture as visualized by scanning electron microscopy. Multidrug-resistance efflux pump gene expression was also analyzed by quantitative reverse transcription PCR. Although the mdrL, mdrT, and lde genes of L. monocytogenes and the mepA gene of MRSA were upregulated, there was no significant difference that indicated an attempt by the efflux pumps to discharge PCEE. MRSA norA expression and abcA expression were significantly downregulated (p < 0.05). A possible mechanism for PCEE may be to cause an energy depletion, either by inhibiting adenosine triphosphate binding or by disturbing the proton gradient, resulting in membrane damage.

Salmonella Contamination in Ready-to-Eat Tilapia Sashimi Processing Plants.

In this study, we investigated contamination rates and possible contamination routes of Salmonella in two typical tilapia sashimi processing plants in Taiwan. We found that the overall isolation rate was 5.0% ( n = 61), from a total of 1,218 samples collected in a year from different processing sections (freezing, scaling and bleeding, visceral removal, washing and disinfection, and packaging) and from different operating times (before processing and 3 and 6 h after processing began). In plant A, which is a relatively well-operated plant compared with plant B, Salmonella was only found in the freezing, scaling and bleeding, and visceral removal sections, with isolation rates ranging from 0 to 9.3%. No Salmonella was identified in the final ready-to-eat products at plant A. In plant B, Salmonella was found in all sections and in the final products, with isolation rates ranging from 4.6 to 36.1%. Regarding the processing times, the contamination rates increased significantly ( P < 0.05) 3 h after processing began in plant B. Among the isolates, 10 serotypes were detected, some of which are commonly observed in human salmonellosis cases in Taiwan, indicating a risk of zoonoses. However, only four isolates showed antimicrobial resistance in the current study. With molecular subtyping, we observed accumulated and persistent Salmonellae contamination patterns in plant B. These results suggest that inadequate sanitation impairs the foodborne pathogen control program in tilapia sashimi plants.

Comparison of high pressure and high temperature short time processing on quality of carambola juice during cold storage.

This study validated high hydrostatic pressure processing (HPP) for achieving greater than 5-log reductions of Escherichia coli O157:H7 in carambola juice and determined shelf life of processed juice stored at 4 °C. Carambola juice processed at 600 MPa for 150 s was identified capable of achieving greater than 5.15-log reductions of E. coli O157:H7, and the quality was compared with that of high temperature short time (HTST)-pasteurized juice at 110 °C for 8.6 s. Aerobic, psychrotrophic, E. coli/coliform, and yeasts and moulds in the juice were reduced by HPP or HTST to levels below the minimum detection limit (< 1.0 log CFU/mL), and showed no outgrowth after refrigerated storage of 40 days. There were no significant differences in pH and titratable acidity between the untreated, HPP, and HTST juices. However, HTST treatment significantly changed the color of juice, while no significant difference was observed between the control and HPP samples. HPP and HTST treatments reduced the total soluble solids in the juice, but maintained higher sucrose, glucose, fructose, and total sugar contents than untreated juice. The total phenolic and ascorbic acid contents were higher in juice treated with HPP than untreated and HTST juice, but there was no significant difference in the flavonoid content. Aroma score analysis showed that HPP had no effect on aroma, maintaining the highest score during cold storage. The results of this study suggest that appropriate HPP conditions can achieve the same microbial safety as HTST, while maintaining the quality and extending the shelf life of carambola juice.

Influence of high-pressure processing on the generation of γ-aminobutyric acid and microbiological safety in coffee beans.

BACKGROUND: The aim of this study was to investigate the influence of high-pressure processing (HPP) on γ-aminobutyric acid (GABA) content, glutamic acid (Glu) content, glutamate decarboxylase (GAD) activity, growth of Aspergillus fresenii, and accumulated ochratoxin A (OTA) content in coffee beans. RESULTS: The results indicated that coffee beans subjected to HPP at pressures ≥50 MPa for 5 min increased GAD activity and promoted the conversion of Glu to GABA, and showed a significantly doubling of GABA content compared with unprocessed coffee beans. Additionally, investigation of the influence of HPP on A. fresenii growth on coffee beans showed that application ≥400 MPa reduced A. fresenii concentrations to <1 log. Furthermore, during a 50-day storage period, we observed that a processing pressure of 600 MPa completely inhibited A. fresenii growth, and on day 50 the OTA content of coffee beans subjected to processing pressures of 600 MPa was 0.0066 µg g-1 , which was significantly lower than the OTA content of 0.1143 µg g-1 in the control group. CONCLUSION: This study shows that HPP treatment can simultaneously increase GABA content and inhibit the growth of A. fresenii, thereby effectively reducing the production and accumulation of OTA and maintaining the microbiological safety of coffee beans. © 2018 Society of Chemical Industry.

Effect of high-pressure processing and thermal pasteurization on overall quality parameters of white grape juice.

BACKGROUND: The aim of this study was to investigate the microbial levels, physicochemical and antioxidant properties and polyphenol oxidase (PPO) and peroxidase (POD) activities as well as to conduct a sensory analysis of white grape juice treated with high-pressure processing (HPP) and thermal pasteurization (TP), over a period of 20 days of refrigerated storage. RESULTS: HPP treatment of 600 MPa and TP significantly reduced aerobic bacteria, coliform and yeast/mold counts. At day 20 of storage, HPP-600 juice displayed no significant differences compared with fresh juice in terms of physicochemical properties such as titratable acidity, pH and soluble solids, and retained less than 50% PPO and POD activities. Although significant differences were observed in the color, antioxidant contents and antioxidant capacity of HPP-treated juice, the extent of these differences was substantially lower than that in TP-treated juice, indicating that HPP treatment can better retain the quality of grape juice. Sensory testing showed no significant difference between HPP-treated juice and fresh juice, while TP reduced the acceptance of grape juice. CONCLUSION: This study shows that HPP treatment maintained the overall quality parameters of white grape juice, thus effectively extending the shelf life during refrigerated storage. © 2016 Society of Chemical Industry.

Prevalence and Persistence of Listeria monocytogenes in Ready-to-Eat Tilapia Sashimi Processing Plants.

A 2-year study was performed at two ready-to-eat tilapia sashimi processing plants (A and B) to identify possible routes of contamination with Listeria monocytogenes during processing. Samples were collected from the aquaculture environments, transportation tanks, processing plants, and final products. Seventy-nine L. monocytogenes isolates were found in the processing environments and final products; 3.96% (50 of 1,264 samples) and 3.86% (29 of 752 samples) of the samples from plants A and B, respectively, were positive for L. monocytogenes . No L. monocytogenes was detected in the aquaculture environments or transportation tanks. The predominant L. monocytogenes serotypes were 1/2b (55.70%) and 4b (37.97%); serotypes 3b and 4e were detected at much lower percentages. At both plants, most processing sections were contaminated with L. monocytogenes before the start of processing, which indicated that the cleaning and sanitizing methods did not achieve adequate pathogen removal. Eleven seropulsotypes were revealed by pulsed-field gel electrophoresis and serotyping. Analysis of seropulsotype distribution revealed that the contamination was disseminated by the processing work; the same seropulsotypes were repeatedly found along the work flow line and in the final products. Specific seropulsotypes were persistently found during different sampling periods, which suggests that the sanitation procedures or equipment used at these plants were inadequate. Plant staff should improve the sanitation procedures and equipment to reduce the risk of L. monocytogenes cross-contamination and ensure the safety of ready-to-eat tilapia products.

Characterization of Gordonia sp. strain CC-NAPH129-6 capable of naphthalene degradation.

A naphthalene-degrading isolate able to utilize naphthalene as a sole carbon source was identified as Gordonia sp. CC-NAPH129-6. Here a detail characterization of the naphthalene catabolic genes present in this strain was conducted. In nar region four structural genes (narAa, narAb, narB, narC), two regulatory genes (narR1, narR2), a rubredoxin encoding gene (rub1) and a gene (orf7) with unknown function were obtained. When compared with most of the members within naphthalene-degrading Rhodococcus, these naphthalene catabolic genes in strain CC-NAPH129-6 were organized into an operon-like gene cluster and present in the same order. This naphthalene gene cluster located in a 97-kb small plasmid of strain CC-NAPH129-6, as can be seen from the PFGE and Southern blot hybridization data. Besides, a partial transposase sequence containing an IS element structure with 12-nt inverted repeat at both ends was found, which was flanked by direct repeats downstream the narC gene in strain CC-NAPH129-6. This novel transposase gene sequence was unlike to the transposase sequence found between narR2 and rub1 genes in Rhodococcus opacus R7. The comparative analyses of the naphthalene catabolic genes, 16S rRNA and gyrB gene present in strain CC-NAPH129-6 and naphthalene-degrading Rhodococcus species imply that the naphthalene catabolic genes in strain CC-NAPH129-6 might be horizontally transferred from Rhodococcus members. This is the first report demonstrating that naphthalene catabolic genes organized into an operon-like gene cluster in the genus Gordonia, and this might provide evidence of the importance of this actinobacterial lineage in the bioremediation of oil-contaminated soils.

Incidence and persistence of Listeria monocytogenes in the catfish processing environment and fresh fillets.

Incidence of Listeria spp. in whole raw catfish, catfish fillets, and processing environments from two catfish processing facilities was determined in August 2008 and August 2009. Thirty-nine (18.4%) of 212 samples collected in August 2008 were positive for Listeria monocytogenes. Prevalences of Listeria species L. innocua and L. seeligeri-L. welshimeri-L. ivanovii were 11.3 and 23.6%, respectively. Of 209 samples collected in August 2009, 12.4% were positive for L. monocytogenes, 11% for L. innocua, and 19.6% for L. seeligeri-L. welshimeri-L. ivanovii. No Listeria grayi was detected in any of the samples. L. monocytogenes was not found in catfish skins and intestines, but was detected in catfish fillets, on food contact surfaces, and on non-food contact surfaces with frequencies of 45.0, 12.0, and 11.1%, respectively. In August 2008 isolates, serotypes 1/2b (62.2%) and 3b (15.6%) were frequently isolated, whereas the majority of the August 2009 isolates (92.3%) were serotype 1/2b. Genotyping analyses revealed that some genotypes of L. monocytogenes isolates were detected in one facility even after a year, but no persistence of L. monocytogenes was observed in the other facility. In addition, some L. monocytogenes isolates from fresh fillets showed genotypes that were either identical, or more than 90% similar, to those of L. monocytogenes isolates from food contact surfaces in the processing lines. The results of this study suggest that processing environment rather than whole raw catfish is an important source of L. monocytogenes contamination in the catfish fillets. These results should assist the catfish industry to develop better control and prevention strategies for L. monocytogenes.

Prevalence and contamination patterns of Listeria monocytogenes in catfish processing environment and fresh fillets.

Catfish skins, intestines, fresh fillets, processing surfaces at different production stages, chiller water and non-food contact surfaces were sampled for Listeria monocytogenes and other Listeria species. Among 315 samples, prevalence of L. monocytogenes, Listeria innocua and a group of Listeria seeligeri-Listeria welshimeri-Listeria ivanovii was 21.6, 13.0 and 29.5%, respectively. No Listeria grayi was detected in this survey. While no L. monocytogenes strains were isolated from catfish skins and intestines, the strains were found with a frequency of 76.7% in chilled fresh catfish fillets and 43.3% in unchilled fillets. L. monocytogenes and Listeria spp. were also detected in fish contact surfaces such as deheading machine, trimming board, chiller water, conveyor belts at different stages, and fillet weighing table. Among L. monocytogenes, 1/2b (47.0%), 3b (16.0%) and 4c (14%) were the predominant serotypes isolated, whereas 4b, 4e, 1/2c and 1/2a were detected at much lower frequencies. Genotype analyses of L. monocytogenes isolates using serotyping, pulsed-field gel electrophoresis (PFGE) and enterobacterial repetitive intergenic consensus (ERIC)-PCR revealed that chiller water represented an important contamination source of L. monocytogenes in the chilled catfish fillets of two processing facilities, whereas fillet weighing table significantly contributed to the catfish fillet contamination of the third facility. This study suggests that L. monocytogenes contamination in the processed catfish fillets originates from the processing environment, rather than directly from catfish. Results from this study can aid the catfish industry to develop a plant-specific proper cleaning and sanitation procedure for equipment and the processing environment designed to specifically target L. monocytogenes contamination.