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Human norovirus (HuNoV) is the leading causative agent of foodborne outbreaks and is associated with the second most prevalent cause of waterborne infections in the United States. The goal of this research was to investigate the antiviral activity of chitosan microparticles (CM) against HuNoV GII.4 Sydney and its cultivable surrogate, Tulane virus (TuV), in suspensions mimicking fecally-contaminated water. CM was prepared by crosslinking chitosan molecules with sodium sulfate, and then its anti-noroviral activity was assessed using infectivity assay on TuV and RT-qPCR on TuV and HuNoV. A 3% CM suspension in PBS (pH 7.2) showed binding to TuV particles but with a negligible impact on virus infectivity (p>0.05). TuV and HuNoV suspended in fecal suspensions showed a 1.5-log10 reduction in genomic copies per ml following a 10-min contact time (p

Foodborne and zoonotic viral pathogens are responsible for substantial morbidity and mortality worldwide. These viruses can be transmitted through foods such as dairy products to humans and cause several acute and chronic diseases. This study aimed to investigate the prevalence and profile of different foodborne and zoonotic viruses in raw cow milk samples. We collected 492 raw cow milk samples from local dairy markets in Qazvin, Iran. Then we evaluated the presence of hepatitis A virus, noroviruses, rotavirus, astrovirus, bovine leukaemia virus (BLV) and tick-borne encephalitis virus (TBEV) in samples using conventional and nested reverse transcription-polymerase chain reaction methods. We found that 34.95, 7.72, 25.81, 14.63, 66.86, 12.80 and 21.34% of raw milk samples were contaminated with norovirus GI, norovirus GII, hepatitis A virus, rotavirus, astrovirus, BLV and TBEV viruses, respectively. Interestingly, the samples collected from the city's south area revealed a higher prevalence of foodborne and zoonotic viruses. Astrovirus and its combination with norovirus GI were the most prevalent virus profiles. Also, the highest correlations were observed among the presence of rotavirus and hepatitis A viruses (0.36) and TBEV and norovirus GII (0.31). Considering the prevalence rate and virus profiles of different foodborne and zoonotic viruses in raw milk samples, hygiene practices and the pasteurization process are strongly suggested to be conducted throughout the cow milk production chain and in dairy industries to prevent infections with these pathogens.

Tuna and mahi-mahi are two major fish species responsible for histamine poisoning. This research developed a rapid ultra-high performance liquid chromatography (UHPLC) method to determine amino acids, histamine, and other biogenic amines that can act as co-indicators of histamine poisoning in tuna and mahi-mahi. The modified UHPLC method could simultaneously determine four biogenic amines and 10 major free amino acids in mahi-mahi (Coryphaena hippurus) and yellowfin tuna (Thunnus albacares) within 17.5 min. This UHPLC method showed good linear response, sensitivity, resolution, recovery, repeatability, and number of theoretical plates. The concentrations of detected amino acids, biogenic amines in mahi-mahi and tuna, and their relationships with fish spoilage grade were determined by this UHPLC method. The developed UHPLC method is a rapid and accurate way to monitor quality changes of mahi-mahi and tuna by inspecting the changes of amino acids and biogenic amines.

Alcohols, aldehydes, ketones, amines, and sulfur compounds are essential aroma compounds related to fish flavor and spoilage. Gas chromatography-mass spectrometry (GC-MS) is an instrument that is widely used to identify and quantify volatile and semi-volatile compounds in fish products. In this research, a simple and accurate GC-MS method was developed to determine the aroma profile of mahi-mahi and tuna for chemical indicators of spoilage. In the developed GC-MS method, trichloroacetic acid (TCA) solution was used to extract analytes from homogenized fish samples. The purge and trap system was used for sample introduction, and the GC-MS with an RTX-Volatile Amine column was able to separate compounds without a derivatization procedure. The created purge and trap gas chromatography-mass spectrometry (PT-GC-MS) method could identify and quantify twenty aroma compounds in mahi-mahi (Coryphaena hippurus) and 16 volatile compounds in yellowfin tuna (Thunnus albacares) associated with fish spoilage. The amines (dimethylamine, trimethylamine, isobutylamine, 3-methylbutylamine, and 2-methylbutanamine), alcohols (2-ethylhexanol, 1-penten-3-ol and isoamyl alcohol, ethanol), aldehydes (2-methylbutanal, 3-methylbutanal, benzaldehyde), ketones (acetone, 2,3-butanedione, 2-butanone, acetoin), and dimethyl disulfide strongly statistically correlated with poorer quality tuna and mahi-mahi and were considered as the key spoilage indicators. Practical Application A simplified and rapid purge and trap gas chromatography-mass spectrometry (PT-GC-MS) method developed in this research was able to identify and quantify important spoilage compounds in mahi-mahi and yellowfin tuna. This method is an efficient analytical method for determining volatile profiles of fish samples for industry analytical labs or the government. The identified analytical quality markers can be used to monitor the spoilage level of tuna and mahi-mahi.

Tuna (Thunnus albacares) and mahi-mahi (Coryphaena hippurus) are two major fish species responsible for scombroid poisoning in the United States. The purpose of this research was to develop a low-cost and easily operated colorimetric strip method for the rapid determination of spoilage degree via amine response in mahi-mahi and tuna. The color strip method was developed by investigating different types of dyes, filter papers, sample volume, water bath temperature, and other parameters. Ultimately rose bengal and bromophenol blue (BPB) dyes were chosen. These two dyes produced standard curves with good linearity (0-50 mg/L for the total biogenic amines) and uniformity of color change. The r2 values for the standard curves of the rose Bengal and BPB were 0.9535 and 0.8883, respectively. Significant positive Pearson correlations coefficients (r) between the volatile biogenic amine levels detected by these two colorimetric strip methods with increasing spoilage grade of mahi-mahi (rose bengal: r = 0.8907, p

Human noroviruses are the leading cause of foodborne gastroenteritis worldwide and disease outbreaks have been linked to contaminated surface waters as well as to produce consumption. Noroviruses are extremely stable in water and their presence is being detected with increasing frequency, yet there are no viable methods for reducing norovirus contamination in environmental water. Despite this, there is little knowledge regarding the physical and chemical factors that influence the environmental persistence of this pathogen. This study evaluated the impact of common chemical and physical properties of surface water on the stability of murine norovirus and examined the effect of food-safe chitosan microparticles on infectivity of two human norovirus surrogates. While chemical additives had a minor impact on virus survival, chitosan microparticles significantly reduced infectious titers of both murine norovirus and MS2 bacteriophage.

Alcohols, aldehydes, ketones, amines, and sulfur compounds are essential aroma compounds related to fish flavor and spoilage. Gas chromatography-mass spectrometry (GC-MS) is an instrument that is widely used to identify and quantify volatile and semi-volatile compounds in fish products. In this research, a simple and accurate GC-MS method was developed to determine the aroma profile of mahi-mahi and tuna for chemical indicators of spoilage. In the developed GC-MS method, trichloroacetic acid (TCA) solution was used to extract analytes from homogenized fish samples. The purge and trap system was used for sample introduction, and the GC-MS with an RTX-Volatile Amine column was able to separate compounds without a derivatization procedure. The created purge and trap gas chromatography-mass spectrometry (PT-GC-MS) method could identify and quantify twenty aroma compounds in mahi-mahi (Coryphaena hippurus) and 16 volatile compounds in yellowfin tuna (Thunnus albacares) associated with fish spoilage. The amines (dimethylamine, trimethylamine, isobutylamine, 3-methylbutylamine, and 2-methylbutanamine), alcohols (2-ethylhexanol, 1-penten-3-ol and isoamyl alcohol, ethanol), aldehydes (2-methylbutanal, 3-methylbutanal, benzaldehyde), ketones (acetone, 2,3-butanedione, 2-butanone, acetoin), and dimethyl disulfide strongly statistically correlated with poorer quality tuna and mahi-mahi and were considered as the key spoilage indicators. PRACTICAL APPLICATION: A simplified and rapid purge and trap gas chromatography-mass spectrometry (PT-GC-MS) method developed in this research was able to identify and quantify important spoilage compounds in mahi-mahi and yellowfin tuna. This method is an efficient analytical method for determining volatile profiles of fish samples for industry analytical labs or the government. The identified analytical quality markers can be used to monitor the spoilage level of tuna and mahi-mahi.

Pathogenic enteric viruses are responsible for a wide range of infections in humans, with diverse symptoms. Raw and partially treated wastewaters are major sources of environmental contamination with enteric viruses. We monitored a municipal secondary wastewater treatment plant (New Orleans, LA) on a monthly basis for norovirus (NoV) GI and GII and enterovirus serotypes using multiplex reverse transcription-quantitative PCR (RT-qPCR) and microbial indicators of fecal contamination using standard plating methods. Densities of indicator bacteria (enterococci, fecal coliforms, and Escherichia coli ) did not show monthly or seasonal patterns. Norovirus GII was more abundant than GI and, along with enterovirus serotypes, increased in influent during fall and spring. The highest NoV GI density in influent was in the fall, reaching an average of 4.0 log 10 genomic copies/100 ml. Norovirus GI removal (0.95 log 10 ) was lower than that for GII, enterovirus serotypes, and male-specific coliphages (1.48 log 10 ) or for indicator bacteria (4.36 log 10 ), suggesting higher resistance of viruses to treatment. Male-specific coliphages correlated with NoV GII densities in influent and effluent ( r = 0.48 and 0.76, respectively) and monthly removal, indicating that male-specific coliphages can be more reliable than indicator bacteria to monitor norovirus GII load and microbial removal. Dominant norovirus genotypes were classified into three GI genotypes (GI.1, GI.3, and GI.4) and four GII genotypes (GII.3, GII.4, GII.13, and GII.21), dominated by GI.1 and GII.4 strains. Some of the seasonal and temporal patterns we observed in the pathogenic enteric viruses were different from those of epidemiological observations.

Clostridium difficile infection (CDI) was generally considered as a hospital-associated disease; however, recent community-acquired CDI has raised the concerns regarding the transmission of the pathogen through environmental sources. Limited data are available regarding the presence of C. difficile in food and water. In this study, oysters (Crassostrea virginica) and the harvest water collected from the commercial harvesting areas along the Louisiana Gulf Coast as well as the influent and effluent of a municipal treatment plant in New Orleans, LA were analyzed for toxigenic C. difficile. The bacterium was isolated from 47.37% (9/19) of oysters and 37.5% (3/8) of harvest water samples. Toxigenic C. difficile were also detected in all the wastewater influent and effluent samples. All the isolates harbored the gene tcdB encoding the virulence factor toxin B. Further PCR-ribotyping showed that the C. difficile isolated from the oysters and harvest water differed from the wastewater isolates. However, similar ribotypes were found in oysters and the surrounding harvest water. We found that oysters growing in contaminated water could bioaccumulate toxigenic C. difficile and pose a health risks by serving as a vehicle for the transmission of the pathogen to humans.

Human norovirus (NoV) is the leading cause of acute gastroenteritis worldwide. Persistence on surfaces and resistance to many conventional disinfectants contribute to widespread transmission of norovirus. We examined the efficacy of neutral electrolyzed water (NEW; pH 7) for inactivation of human NoV GII.4 Sydney in suspension (ASTM method 1052-11) and on stainless steel surfaces (ASTM method 1053-11) with and without an additional soil load. The impact of the disinfectant on viral capsid was assessed using reverse transcriptase quantitative PCR (RT-qPCR; with an RNase pretreatment), SDS-PAGE, transmission electron microscopy, and a histo-blood group antigen (HBGA) receptor-binding assay. These studies were done in parallel with those using Tulane virus (TuV), a cultivable human NoV surrogate. Neutral electrolyzed water at 250 ppm free available chlorine produced a 4.8- and 0.4-log 10 reduction in NoV genome copy number after 1 min in suspension and on stainless steel, respectively. Increasing the contact time on surfaces to 5, 10, 15, and 30 min reduced human NoV genomic copies by 0.5, 1.6, 2.4, and 5.0 log 10 and TuV infectious titers by 2.4, 3.0, 3.8, and 4.1 log 10 PFU, respectively. Increased soil load effectively eliminated antiviral efficacy regardless of testing method and virus. Exposure to NEW induced a near complete loss of receptor binding (5 ppm, 30 s), degradation of VP1 major capsid protein (250 ppm, 5 min), and increased virus particle aggregation (150 ppm, 30 min). Neutral electrolyzed water at 250 ppm shows promise as an antinoroviral disinfectant when used on precleaned stainless steel surfaces. IMPORTANCE Norovirus is the leading cause of acute viral gastroenteritis worldwide. Transmission occurs by fecal-oral or vomitus-oral routes. The persistence of norovirus on contaminated environmental surfaces exacerbates its spread, as does its resistance to many conventional disinfectants. The purpose of this research was to evaluate the antinoroviral efficacy of neutral electrolyzed water (NEW), a novel chlorine-based disinfectant that can be used at reduced concentrations, making it more environmentally friendly and less corrosive than bleach. An industrial-scale electrochemical activation device capable of producing relatively stable electrolyzed water at a wide pH range was used in this study. Experiments showed that 250 ppm NEW effectively eliminated (defined as a 5-log 10 reduction) human norovirus GII.4 Sydney (epidemic strain) on clean stainless steel surfaces after a 30-min exposure. Supporting studies showed that, like bleach, NEW causes inactivation by disrupting the virus capsid. This product shows promise as a bleach alternative with antinoroviral efficacy.

Human norovirus (NoV) is the leading cause of foodborne illnesses in the United States. Norovirus is shed in high numbers in the feces and vomitous of infected individuals. Contact surfaces contaminated with bodily fluids harboring infectious virus particles serve as vehicles for pathogen transmission. Environmental stability of NoV and its resistance to many conventional disinfectants necessitate effective inactivation strategies to control the spread of virus. We investigated the efficacy of two commercial disinfectants, hydrogen peroxide (7.5%) and a chlorine dioxide (0.2%)-surfactant-based product using a fogging delivery system against human NoV GI.6 and GII.4 Sydney strains as well as the cultivable surrogate, feline calicivirus (FCV) dried on stainless steel coupons. Log10 reductions in human NoV and FCV were calculated utilizing RNase RT-qPCR and infectivity (plaque) assay, respectively. An improved antiviral activity of hydrogen peroxide as a function of disinfectant formulation concentration in the atmosphere was observed against both GII.4 and FCV. At 12.4 ml/m3, hydrogen peroxide achieved a respective 2.5 ± 0.1 and 2.7 ± 0.3 log10 reduction in GI.6 and GII.4 NoV genome copies, and a 4.3 ± 0.1 log10 reduction in infectious FCV within 5 min. At the same disinfectant formulation concentration, chlorine dioxide-surfactant-based product resulted in a respective 1.7 ± 0.2, 0.6 ± 0.0, and 2.4 ± 0.2 log10 reduction in GI.6, GII.4, and FCV within 10 min; however, increasing the disinfectant formulation concentration to 15.9 ml/m3 negatively impacted its efficacy. Fogging uniformly delivered the disinfectants throughout the room, and effectively decontaminated viruses on hard-to-reach surfaces. Hydrogen peroxide delivered by fog showed promising virucidal activity against FCV by meeting the United States EPA 4-log10 reduction criteria for an anti-noroviral disinfectant; however, fogged chlorine dioxide-surfactant-based product did not achieve a 4-log10 inactivation. Future investigation aimed at optimizing decontamination practices is warranted.

Fish sausage formulation included pale-meat pink salmon (Oncorhynchus gorbuscha), wild salmon oil and extra-virgin coconut oil as lipid sources, wholegrain oats as texturizer, natural spices, salt, brown sugar and sodium polyphosphate. Uncooked and smoked sausages were evaluated for their nutritional value, microbial load, color and texture. Hot smoke processing caused a 6% reduction in the moisture content of the uncooked product (69% w/w), an increase in the product's Allo-Kramer shear value, and a reduction in bacterial counts to less than 2 log cfu/g. In one link serving size, the fat and n-3 polyunsaturated fatty acid contents were 6.45 and 6.89% w/w and 837 and 891 mg for uncooked and smoked sausages, respectively. Salmon sausages are low in fat and a good source of healthy lipids and marine proteins. The products were created to meet consumers’ demand for functional foods that do not contain land-based animal ingredients. Practical Applications Healthy lipids are wanted for everyone's daily diet and have become common in many foods. Application of healthy lipid sources such as fish oil in sausage formulation, a food product generally high in fat, provides a healthy alternative to traditional sausage products. Uncooked and smoked fish sausages were developed using pale-meat pink salmon and wild Alaska salmon oil. These low-fat functional food products contain healthy long-chain n-3 fatty acids, proteins of marine origin only, and wholegrain oats.

Cold-smoked salmon (CSS) is a potentially hazardous ready-to-eat food product due to the high risk of contamination with Listeria monocytogenes and lack of a listericidal step. We investigated the antilisterial property of liquid smokes (LS) against Listeria innocua ATCC 33090 (surrogate to L. monocytogenes) as a potential supplement to vacuum-packaged CSS. A full-strength LS (Code 10-Poly), and three commercially refined fractions (AM-3, AM-10, and 1291) having less color and flavor (lower content of phenols and carbonyl-containing compounds) were tested. In vitro assays showed strong inhibition for all LS except for 1291. The CSS strips were surface coated with AM-3 and AM-10 at 1% LS (vol/wt) with an L-shaped glass rod and then inoculated with L. innocua at 3.5 log CFU/g, vacuum packaged, and stored at 4°C. The LS did not completely eliminate L. innocua but provided a 2-log reduction by day 14, with no growth up to 35 days of refrigerated storage. A simple difference sensory test by 180 untrained panelists showed the application of AM-3 did not significantly influence the overall sensorial quality of CSS. In essence, the application of the refined LS as an antilisterial additive to CSS is recommended.

The biogenic amine (putrescine, cadaverine, histamine and tyramine) content of whole Southern Caspian Kutum (Rutilus frisii kutum) and related bacterial changes (Pseudomonas spp., psychrotrophic and mesophilic counts) were monitored during ice storage for a period of 18 days (0, 3, 6, 9, 12, 15 and 18). Levels of putrescine, cadaverine and bacterial loads increased (P

The biogenic amine content and related bacterial changes (Pseudomonas spp., psychrotrophic and mesophilic counts) in whole farmed rainbow trout (Oncorhynchus mykiss) were monitored during ice storage for 18 days (at 0, 3, 6, 9, 12, 15 and 18 days). Levels of putrescine, cadaverine and histamine, and bacterial loads, increased (P < 0.05) during storage, but tyramine was not detected in any of the tested samples. Concentration of putrescine ranged from 0.4 initially to 8.97 μg/g, and psychrotrophic microorganisms were dominant. The best linear regressions (correlations) were for putrescine and Pseudomonas spp. and psychrotrophs (r = 0.98), and for cadaverine with Pseudomonas spp. (r = 0.82). Putrescine content was a good quality marker. Histamine was detected only at later stages of storage and was therefore less suitable than the other biogenic amines as freshness indicator.

Noroviruses are the most common causative agent of viral gastroenteritis in humans, and are responsible for major foodborne illnesses in the United States. Filter-feeding molluscan shellfish exposed to sewage-contaminated waters bioaccumulate viruses, and if consumed raw, transmit the viruses to humans and cause illness. We investigated the occurrence of norovirus GI and GII and microbial indicators of fecal contamination in the eastern oysters (Crassostrea virginica) and water from commercial harvesting areas along the Louisiana Gulf Coast (January to November of 2013). Microbial indicators (aerobic plate count, enterococci, fecal coliforms, Escherichia coli, male-specific coliphages, and somatic coliphages) were detected at the densities lower than public health concerns. Only one oyster sample was positive for norovirus GII at 3.5 ± 0.2 log10 genomic equivalent copies/g digestive tissues. A stool specimen obtained from an infected individual associated with a norovirus outbreak and the suspected oysters (Cameron Parish, La., area 30, January 2013) were also analyzed. The norovirus strain in the stool belonged to GII.4 Sydney; however, the oysters were negative and could not be linked. In general, no temporal trend was observed in the microbial indicators. Low correlation among bacterial indicators was observed in oysters. Strongest correlations among microbial indicators were observed between enterococci and fecal coliforms (r = 0.63) and between enterococci and E. coli (r = 0.64) in water (P0.05); however, weak correlations were found in oysters (r0.45) and between oysters and harvest water (r ≤ 0.36, P0.05). Our results emphasize the need for regular monitoring of pathogenic viruses in commercial oyster harvesting areas to reduce the risks of viral gastroenteritis incidences.

The objective of this study was to determine important chemical characteristics of a full-strength liquid smoke, Code 10-Poly, and three refined liquid smoke products (AM-3, AM-10 and 1291) commercially available (Kerry Ingredients and Flavors, Monterey, TN). The pH of the products were significantly different (P 0.05) and ranged from 2.3 (Code 10-Poly) to 5.7 (1291). The pH was inversely correlated with titratable acidity (R (2) = 0.87), which was significantly different (P 0.05) among products ranging from 10.3% acetic acid (Code 10-Poly) to 0.7% acetic acid (1291). Total phenol content was quantified using the Gibbs reaction; the only liquid smoke containing appreciable level of phenolic compounds was Code 10-Poly at 3.22 mg mL(-1). Gas chromatography-mass spectrometry (GC-MS) analysis of liquid smoke dichloromethane extracts revealed that carbonyl-containing compounds were major constituents of all products, in which 1-hydroxy-2-butanone, 2(5H)-furanone, propanal and cyclopentenone predominated. Organic acids were detected by GC-MS in all extracts and correlated positively (R (2) = 0.98) with titratable acidity. The GC-MS data showed that phenolic compounds constituted a major portion of Code 10-Poly, and were detected only in trace quantities in 1291. The refined liquid smokes had lighter color, lower acidity, and reduced level of carbonyl-containing compounds and organic acids. Our study revealed major differences in pH, titratable acidity, total phenol content, color and chemical make-up of the full-strength and refined liquid smokes. The three refined liquid smoke products studied have less flavor and color active compounds, when compared with the full-strength product. Furthermore, the three refined products studied have unique chemical characteristics and will impart specific sensorial properties to food systems. Understanding the chemical composition of liquid smokes, be these refined or full-strength products, is an important step to establish their functions and appropriate use in food systems.