Vibrio parahaemolyticus, a Gram-negative, salt-loving bacterium common in marine environments, is the leading cause of acute hepatopancreatic necrosis, also known as “early death syndrome,” in aqua culture, and is responsible for a significant number of foodborne illnesses in humans.

Over the past two decades, the bacteria has led to a significant rise of infections in humans, more so than other foodborne pathogens. These infections primarily result from consuming raw fish and seafood, and particularly, shellfish.

Climate change, causing rising ocean temperatures and ocean acidification, has resulted in increased abundances of Vibrio parahaemolyticus in oceans worldwide. In fact, the most recent FoodNet annual report indicates that the overall incidence in 2021 rose by 45.5% when compared with the annual incidence from 2016 to 2018.

Traditional detection methods for bacteria are labor intensive and time consuming, falling short of the need for accurate, rapid, and convenient detection required by food safety supervision and food enterprises; however, researchers in Shanghai, China, have developed a point-of-care detection method that allows for the quick and sensitive identification of the bacteria in seafood.

This new method uses advanced techniques called recombinant polymerase amplification (RPA) and the CRISPR/Cas12a system, along with a test strip. The method provides a low-cost, simple, and visually clear way to quickly detect Vibrio parahaemolyticus in seafood.

The researchers note that RPA-CRISPR/Cas12a-ICS can detect Vibrio parahaemolyticus in salmon sashimi at extremely low levels, as little as 154 CFU/g, without needing to enrich the sample first. “Our innovative detection platform represents a significant advancement in the rapid and sensitive detection of Vibrio parahaemolyticus, proving especially valuable for ensuring seafood safety and preventing public health crises,” corresponding author Haijuan Zeng, leader of the Biotechnology Research Institute at the Shanghai Academy of Agricultural Sciences, said in a prepared statement.

Zeng, who designed and performed the experiments and analyzed the data, explained that by using this platform, Vibrio parahaemolyticus can be detected in approximately 30 minutes, with a limit of detection of 250 copies/μL for plasmid samples and 140 CFU/mL for bacteria. The platform has been validated with artificially contaminated food samples and various clinical isolates.

Furthermore, in the report, the researchers noted that adjusting the crRNA sequences could enable the identification of various other targets, allowing the optimized ssDNA concentration to be used for detecting different targets. Therefore, the RPA-CRISPR/Cas12a-ICS platform could be employed to detect foodborne pathogens linked to humans, adulterated foods, and even viruses.

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Even prior to President Trump’s 2020 executive order expanding offshore fish farming in the U.S., aquaculture was being touted as the future of sustainable fishing. As global fish stocks continue to shrink due to overfishing, fish and shellfish farming seems like an obvious move. It’s a far more efficient way to raise meat for protein than farming chickens, pigs, and cows, which currently occupy more than 37% of the earth’s habitable land. Done right, aquaculture can help maintain healthy waterways and boost jobs and economies in the areas that serve aquacultural regions.

The history of aquaculture stretches back thousands of years. In North America, indigenous people of the Pacific Northwest region historically farmed herring eggs, octopus, clams, and salmon, while indigenous Hawaiians developed freshwater and intertidal fish ponds. Chinese fish farmers domesticated carp around 3500 BCE. Yet, if aquaculture is the past and the future, it’s also the present: Currently, half of the world’s fish and seafood is raised through aquaculture and, according to a 2023 whitepaper from the World Economic Forum, the global demand for those foods is expected to double by 2050.

Following President Trump’s executive order and a bipartisan bill supporting offshore fish farming in the House of Representatives, many American companies have been willing to bet on, and invest in, fish farming. In 2017, 90% of the seafood eaten by Americans came from other countries, and many feel it’s time for American consumers to eat fish and seafood produced and farmed here. Following the 2020 executive order, the Army Corps of Engineers issued permits for aquaculture structures in federal waters.

While the field of fish and seafood farming may be ancient, food safety experts agree that it must be held to exacting modern standards and regulation.

Aquaculture Systems

There are dozens of different approaches to aquaculture. For many, “fish farming” calls to mind offshore net pens—net-cages floating in open water—however, this is only one type of aquacultural technology.

Rome, Italy-based Matthias Halwart, PhD, is the sustainable aquaculture team leader for the Food and Agriculture Organization of the United Nations. He says that, given the wide variety of aquaculture possibilities, the choice of system and approach must be decided according to, among other things, the species being grown, the local environment, and the investment available to farmers. “Finfish can be grown in floating cages [net pens] in freshwater lakes and rivers, brackish estuaries, or in marine coastal or offshore areas,” he says. “Mussels are grown attached to long ropes in the sea connected to floating buoys. Seaweed is also grown on long lines. Pond culture is the most widely practiced method of aquaculture, and ranges from low-intensity green water ponds with low stocking density, using fertilizer to encourage algae and plankton to grow as feed for fish, to highly intensive with formulated feed and aeration using paddle wheels or air blowers.”

Additionally, there are more technical set-ups known as recirculating aquaculture systems (RAS). “With these highly technical systems,” Dr. Halwart says, “the operators are able to manage the water temperature, water quality, and filtration, and control the chemical properties of the water through monitoring. [They] can achieve very intensive levels of production. A version of RAS can be connected to hydroponic vegetable production, called aquaponics, in which the waste water from the fish can serve as fertilizer for the plants, while, at the same time, the plants filter the water for the fish.” Dr. Halwart adds that each of these farming systems has benefits and disadvantages, and that a good system matches the needs of the farmer and the realities of the local conditions.

Approaches to aquaculture, Dr. Halwart says, break down into water-based systems (such as cages and pens), land-based systems (such as rain-fed ponds, irrigated systems, tanks, and raceways), recycling systems (designed to recirculate water in large, closed vessels), and integrated farming systems that pair aquaculture with livestock or crop farming. Different seafood and fish require different aquaculture approaches. Fish are raised in ponds, molluscs are grown in a variety of styles both on and off the seafloor, crustaceans are raised in ponds and concrete raceways, and seaweeds and minor invertebrates are farmed across a variety of systems.

Aquaculture expert Carole Engle, PhD, former executive editor of the Journal of the World Aquaculture Society and adjunct faculty at Virginia Tech’s Virginia Agricultural Research and Extension Centers in Hampton, Va., says that in the United States, aquaculture styles are determined by the popularity of the fish that’s cultivated. “Aquaculture is incredibly diverse and every aquatic animal or plant has to be raised in a different way because the biology is so different,” she adds (see “Top 5 Most-Frequently Farmed and Fished Seafood in the U.S.,”  below).

Aquaculture and Food Safety

Michael Ciaramella, PhD, is Seafood Safety and Technology Specialist at the Sea Grant organization’s Cornell Cooperative Extension in Stony Brook, NY. Due to the breadth of approaches to aquaculture, he finds it hard to generalize about food safety across the sector.

He does note one potential hazard that separates aquaculture from wild fish: pharmaceuticals. “There are only a few drugs approved for use in food fish,” he says. “Strict protocols for their use are in place to ensure they do not impact the safety of the fish as food. Seafood processors must address this potential hazard in their food safety plans and assure that, if aquaculture drugs are used, they are used in accordance with current requirements and best practices.”

Beyond that, Dr. Ciaramella says that food safety concerns in aquaculture are similar to food safety challenges facing open-water fisheries. He adds that environmental contaminants (e.g., heavy metals, herbicides, and pesticides) and natural toxins (those produced by harmful algal blooms) can be an issue for both farmed and wild fish; however, he says that both contaminants and natural toxins can be controlled at the farm level by knowing the potential hazards associated with the various water bodies, and only growing food fish in water bodies with little to no known contamination, or sourcing waters suitable for aquaculture production in land-based systems.

But, this is more complicated than it may sound. Dr. Ciaramella specifies that contaminants pass into species through the things they eat, including fish meal composed of smaller bait fish. Consequently, he says, it’s integral that farmed fish receive high-quality feed that has been tested for contamination. The same wild bait fish used for fish meal are also consumed by wild-caught fish, meaning that contamination in bait fish threatens wild-caught and farmed fish.

One risk particular to fish-farming feeds, he says, is contamination by terrestrial ingredients. “If there are non-marine alternative proteins and plant-based components to the feeds, these could contribute additional contaminants and be a potential hazard unique to farmed species when pelleted feeds are used.”

American Aquaculture

The good news about American aquaculture, says Dr. Engle, is that its systems are set up to present fewer food safety challenges than in other parts of the world. While regulations vary from state to state, aquaculture is overseen by FDA—and, in the case of catfish, by USDA. Catfish is such a big business, Dr. Engle says, that the industry approached congress to request that their production facilities be overseen by USDA’s Food Safety and Inspection Service, rather than just by FDA. This means that each catfish processing plant has an in-house inspector.

The same is not true of other aquacultures, though Dr. Engle stresses that because earthen ponds, raceways, and above-ground tanks work with captive water from well-tested groundwater aquifers they reuse for 10 to 15 years, food safety concerns associated with open-water farming are not present. In particular, fish raised in ponds and sold live face few of the food safety challenges associated with processed fish.

Additionally, adds Dr. Halwart, aquatic animals feeding low in the food chain, such as carp or tilapia, typically have fewer problems with accumulation of toxins. “Disease outbreak is usually associated with intensity of farming; the more intensively you produce, the more careful you have to be with health management,” he adds.

Shellfish food safety, however, is both easier to control in some ways, and harder in others. Bill Walton, PhD, is the Acuff Professor of Marine Science and Shellfish Aquaculture and program coordinator at William and Mary’s Virginia Institute of Marine Science in Gloucester Point. “You don’t feed [shellfish],” he says, “which also has the implication you don’t medicate them. You are relying on the food in that environment, which also means when you think about sustainability, I can’t grow more shellfish in an acre than that acre naturally supports.”

The bad news about shellfish is that, because they’re sometimes not cooked, any pathogen that gets into an oyster may be passed directly to the consumer. In many cases, shellfish producers have been able to mitigate those risks through close scrutiny of water. “The areas available to harvest have to be regularly sampled,” Dr. Walton says. “Typically they’re okay to harvest from unless something happens—something as simple as a certain number of inches of rainfall—then we close. We don’t wait for the lab; we don’t wait for somebody to go collect samples. You can just look at the rain gauge and say, preemptively, ‘We no longer meet the conditions to be open right now, so we’re going to close.’ The model has been that it’s easy to close, and the burden of proof is on reopening, and that’s worked pretty well.”

In that sense, Dr. Walton says that regulation has solved the challenge of pollution in shellfish. Unfortunately, bacterial contamination is not as easy. “If it were associated with pollution, we would’ve solved it,” he says. But it’s not; bacterial contamination simply occurs in the water, the same water people might enjoy playing in at the beach.

The Cold Chain

Dr. Walton says that the solution for shellfish food safety has been the cold chain, “Having a clear process where everybody along the cold chain has to document this, there are tags that go from harvest all the way to the final consumer that demonstrate who has it, and there’s a time–temperature log that has to be kept. We’ve found if you harvest shellfish, and you get them cold right away, and you keep them cold, that dramatically limits the risks.”

Dr. Halwart agrees—and not just for shellfish, but for aquaculture products generally. He notes, “A strong cold chain—meaning the product is immediately chilled after harvest and remains chilled until consumption—is key for many aquaculture products as well. Value addition activities, such as smoking, drying, curing, fermenting, or salting (and good practices associated with these processes) are also good options and traditionally used when cold chain is not available or doesn’t match the market and culinary traditions of the consumer.”

Sustainability

The sustainability of aquaculture—particularly when compared with meat animals raised on land—is a feature fish-farming advocates often highlight. NOAA lists a number of benefits associated with aquaculture—specifically, marine aquaculture operations typically have smaller carbon footprints and require less land and fresh water. Further, they tend to be more effective at converting feed into protein for human consumption than beef, pork, and poultry.

Yet, in 2020, the journal Global Environmental Change published a report from a team of researchers from universities in Norway, Australia, and Chile that targeted aquaculture certification schemes (doi: 10.1016/j.gloenvcha.2019.102025). Their research found that the leading challenge to aquaculture sustainability was the certification under which aquaculture was defined as “sustainable.” In general, the researchers found that aquaculture sustainability certification systems tended to reflect mainly “environmental and governance indicators, and only display scattered attempts at addressing cultural and economic issues. […] The strong bias implies that these certification schemes predominantly focus on the environmental domain and do not address sustainability as a whole, nor do they complement each other. Sustainability is by definition and by necessity a comprehensive concept, but if the cultural and economic issues are to be addressed in aquaculture, the scope of certification schemes must be expanded.”

Dr. Ciaramella agrees that a truly sustainable operation must be environmentally, socially, and economically sustainable. “Most tend to focus on the environmental aspects of sustainability and neglect the social and economic components,” he says. He also stresses that a closer sustainability challenge to the fish farm itself is sourcing protein for farmed fish. “The main protein source for farmed carnivorous fish has historically been fish meal, which relies heavily on the wild capture of small species of fish to produce fish meal and, ultimately, pelleted feeds. There have been many advances in alternative protein technologies moving toward more sustainable feed production. This includes the use of plant- and insect-based proteins.”

These advances have been paired with new production technology systems such as water filtration tools and an aquaculture technique called integrated multitrophic systems, which Dr. Ciaramella says rely on growing multiple species of different trophic levels together to feed off of one another and limit the overall impact on the surrounding ecosystem.

While aquaculture is not completely without food safety concerns, the method offers a valuable source of seafood and supports global food security

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An Israeli food-tech company says it has produced a 3D-printed fish product made with animal cells grown in a laboratory.

Steakholder Foods has partnered with Singapore-based Umami Meats to develop a scalable process for producing structured cultivated fish products using its 3D bio-printing technology and customized bio-inks. The printing and bio-ink customization are steps on the path to commercializing Steakholder Foods’ 3D printer. Unlike fully cultivated meat products that still require incubation and maturation after printing, the grouper fish product is ready to cook after printing.

Since receiving grouper fish cells from Umami, Steakholder Foods is working on the taste and texture of its printed grouper before finalizing a prototype. Umami says that the product mimics the flaky texture of cooked fish.

Umami hopes to bring the fish to market next year, starting in Singapore.

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Plastics continue to be produced at an unprecedented rate. While they may be cheap and convenient, plastics can also take hundreds of years to decompose, and are accumulating at an increasing speed in our environment.

Microplastics, composed of plastics that are 100 nm to 5 mm in size, are a classification of plastic that has either been deliberately manufactured at that size or has degraded from larger pieces of plastic. Plastics have been found almost everywhere in the environment, from the tops of remote mountains to the depths of oceans, with samples even collected from snow in the Arctic. In addition to polluting our air, water, and soil, recent studies have confirmed the presence of unwanted microplastics in common consumer products such as salt and bottled water.

At the University of Mississippi, we are conducting research to help us better characterize and understand the prevalence of microplastic pollution in oyster reefs and other coastal sites in the Mississippi Sound along the Gulf Coast. Through this research, it’s our aim to better understand the prevalence and threat of microplastics in order to better inform our ability to regulate and prevent this emerging containment from further entering our environment and our food chain.

Understanding Microplastics as an Emerging Environmental Contaminant

Microplastics have polluted our environment and are now pervasive in our oceans, lakes, rivers, air, and soil. Our oceans face an acute threat, with an estimated four to 12 million tons of plastic waste entering the oceans every year, posing a serious environmental threat to aquatic species.

Microplastics are damaging our ecosystem and negatively impacting our ocean life, threatening disruption and damage to the digestive tract if ingested. They also raise the risk of entanglement and a host of other negative consequences for aquatic species.

This interaction between microplastics and seafood can also have a negative impact on our food chain, both by contaminating the seafood we eat and by harming seafood populations. The majority of our seafood comes from estuaries and coastal areas, such as oyster reefs. It is in these estuaries and coastal areas that microplastics accumulate, due to the continual input and degradation of plastic litter from rivers and runoff.

Filter feeders like mollusks and oysters (Crassostrea virginica) are particularly vulnerable to microplastic pollution. However, few research papers have investigated the exposure of microplastics in oysters or by oyster reefs.

A Closer Look at Oyster Populations in the Mississippi Sound

Microplastics pose a significant threat to oyster populations, which have already decreased in recent years due to a combination of pollution (e.g., oil spills) and weather events, such as hurricanes and flooding.

To better understand this issue of microplastic prevalence in oyster habitats, we recently conducted a study examining the concentration of microplastic pollution in oyster reefs and other coastal sites in the Mississippi Sound, as well as the impact of freshwater inflows from flooding to these sites. We collected water samples from 10 sites, of which four were directly above oyster reefs.

Recent studies show that oysters nearer urban centers often contain higher concentrations of microplastics, which, given the prevalence of commercial fishing, oil drilling, and shipping ports in the area, implies that the Gulf Coast could be accumulating a considerable number of microplastics.

This was consistent with our findings, which estimated that oysters may be exposed to nearly 24,000 microplastics daily (range ~5,600 to ~36,000), understanding that concentration and filtering rates vary depending on other factors such as site-specific conditions and oyster species. To put this into perspective, humans are estimated to consume anywhere between 39,000 to 52,000 microplastic particles a year, meaning oysters are potentially exposed to half of our annual exposure every day. Overall, the study concluded that seawater along the Mississippi Gulf Coast had higher abundances of microplastics than what was observed in the Mississippi River and its tributaries to coastal areas.

The study also confirmed that estuaries have higher concentrations of microplastics than their riverine inputs, a finding uncovered in other studies as well. Given that the river is continually flushed of plastics, this is not surprising, because the estuary acts as a sink for these plastics, which, over time, degrade to form microplastics.

Research Methods

The microplastics particles that we are analyzing are often so small that they are invisible to the human eye. The number of microplastics actually increases with decreasing size, which calls for sophisticated analytical instrumentation and a robust research approach.

To analyze the abundance of microplastics in these oyster reefs and understand their potential threat, we used three key research methods: “the single one-pot” method for sample preparation, Nile red fluorescence to quantify the collected microplastics samples, and LDIR analysis to identify the major types of plastics in the collected samples. The “single one-pot method” is a novel approach developed by our lab that involves utilizing inexpensive jars, such as mason jars, to collect and prepare water samples for analyses. The advantage of this method is that it minimizes contamination and sample loss, because the sample is processed in the same jar in which it is collected, and the process successfully isolates microplastics needed for analyses.

These microplastic samples were then quantified using Nile red fluorescence detection. Adding a few drops of Nile red dye onto filters with microplastics reveals the exact quantity of microplastics within the samples. To identify the key types and size fractions of plastics in the collected samples, Agilent’s 8700 system was used. This instrument is the first major application of LDIR analyses to determine, characterize, and identify microplastics in natural waters.

A combination of the instrument’s proprietary quantum cascade laser (QCL) with a single-point mercury cadmium telluride (MCT) detector and rapid scanning optics allowed for two effective modes of action. By actioning these techniques, particles are located in the first step, and then information on the size and shape of particles can be obtained. In the second step, a full spectrum is acquired for each particle, while the surrounding areas are ignored. This information is then compared to a spectral database built into the software in a fraction of the time needed with a traditional FTIR system.

The Critical Nature of This Research

Due to mass plastic production, there is now an extensive and increasing amount of microplastics in our environment, and scientific research has not yet uncovered an effective method to entirely remove these particles. Because this is a relatively new threat to the environment, further research needs to take place to understand the true impact of these pollutants, but one thing we do know is that these contaminants do not belong in our environment at all, and certainly not on such a large scale.

Existing research already supports the fact that plastic contaminants pose a threat to aquatic organisms, and it’s very possible that these plastic particles also pose a threat to our own health given the rate that they are entering our environment and our food chain. The research we are conducting at the University of Mississippi is essential to filling in blanks on a known threat to our ecosystem. There is hope that this research could go on to inspire further needed research on the topic and inform our understanding of the exact nature of this threat to human and animal health.

Such research has the potential to guide policymakers in developing needed strategies to control and mitigate this environmental threat and provide evidence to regulate the use of our plastic.

One caveat, however, is that different labs around the world performing microplastics research adopt different testing approaches. These variations may hinder the accurate determination of the fate of microplastics and its global distribution across our oceans. Therefore, global harmonization and standardization of monitored microplastics testing methods is essential if we are to transform this investigative research into routine environmental screening procedures.

Our research has provided evidence of the abundance of microplastics in an area of critical ecological and commercial importance in our region. It’s important that we continue investing in new ways to research and understand contaminants like microplastics if we are going to make progress in the fight against them.

The future of microplastics testing needs widely accepted reference materials so that researchers can use them to assess methods and harmonize plastic pollution research. Also, additional scrutiny needs to be placed on analyses of fibers, which are often excluded from studies or not integrated within a broader context. Lastly, work needs to continue on the complex interactions between microplastics and other marine pollutants to address ongoing questions regarding the potential health risk associated with microplastics.

Academia, along with the life sciences industry, policy makers, and the public must all work together to reduce this type of pollution in our environment and to find new and innovative ways to remove that which is already there. It’s in our interest and that of future generations that we act now.

References for this article are available upon request.

Dr. Cizdziel is an associate professor of chemistry and biochemistry at the University of Mississippi. Reach him at email cizdziel@olemiss.edu.

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The pandemic is putting a strain on the seafood industry, according to a new report focused on the impact of COVID-19 on U.S. fisheries. The study investigators suggest that American fishmongers may struggle without additional government aid.

The study, published in Fish and Fisheries, found that monthly fresh seafood exports declined up to 43% compared with 2019, while monthly imports fell up to 37%, and catches dropped 40% some months. Over the first six months of 2020, total U.S. seafood exports are down 20%, and imports are down 6%, compared with the same period last year. Further losses are likely as restrictions increase to address COVID-19.

“Seafood has been hit harder than many other industries because many fisheries rely heavily on restaurant buyers, which dried up when the necessary health protocols kicked in,” says lead author Easton White, PhD, of the University of Vermont in Burlington. “Restaurants represent about 65% percent of U.S. seafood spending, normally.” For context, more than one million U.S. seafood workers regularly produce more than $4 billion in annual exports, much of which is processed overseas and imported back to the U.S.

In January 2020, demand for American imports plummeted as lockdowns began in China. Starting in March, web searches for U.S. seafood restaurants fell more than 50% and foot traffic at seafood markets decreased 30%.

Additional Funding Needed for the Seafood Industry

Aid for fisheries has been slow, partly because pandemics are not currently considered valid reasons for a fishery failure or disaster under current law. The CARES act has authorized $300M for the sector. Even with increased demand for seafood delivery, which surged 460% for Google searches from March to April, some producers may not be able to recover without government assistance.

“Seafood is a seasonal business,” adds White. “If you have a March to June season, and can’t get funds until next year, you might have to quit. Support from policymakers will decide which producers can survive.”

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Bipartisan support for The Advancing the Quality and Understanding of American Aquaculture (AQUAA) Act, a new bill introduced in the U.S. Senate by Senators Roger Wicker (R-MS), Brian Schatz (D-HI) and Marco Rubio (R-FL), is looking to expand offshore aquaculture in hopes of providing stable economic opportunities for Americans while preserving ecosystems and improving public health. The proposed bill, which has companion legislation in the U.S. House, would support development of an offshore aquaculture industry in the U.S. to increase the production of sustainable seafood and establish new economic opportunities in federal waters.

The National Oceanic and Atmospheric Administration reports that more than 80% of the seafood eaten by Americans is imported, as U.S. fishers and aquaculture operations have not been able to keep up with the country’s demand for fish and other seafood.

“Aquaculture is the fastest growing food production sector, but the U.S. lacks a comprehensive, nationwide system for permitting in federal waters,” said Sen. Wicker when announcing the bill. “This deficiency prevents the development of aquaculture farms, leading to more seafood imports. Our legislation would establish national standards for offshore aquaculture, enabling U.S. producers to create jobs and meet the growing demand for fresh, local seafood.”

Bill DiMento, president of Stronger America Through Seafood, said the expansion of domestic offshore aquaculture is a win–win for American communities nationwide. “With our extended coastline, expansive ocean resources, skilled labor force, superior technology, and ample feed sources, the U.S. has the potential to be a global leader for aquaculture production,” he says. “However, our potential will remain untapped unless and until federal action is taken to clarify the permitting process.”

If passed, the AQUAA Act would establish national standards for offshore aquaculture and clarify a regulatory system for the development of aquaculture in the U.S. exclusive economic zone. The legislation would also establish a research and technology grant program to fund innovative research and extension services focused on improving and advancing sustainable domestic aquaculture.

Opponents argue that offshore fish farms compete with commercial fishing interests, could create pollution due to fish waste, and may possibly spread diseases to wild fish populations. Additionally, they would rather congress support sustainable seafood production with local fishermen and businesses.

Among those who came out opposed to offshore aquaculture are the Gulf Fishermen’s Association, the Center for Food Safety, and Healthy Gulf. “Industrial finfish aquaculture facilities harm wild ecosystems … and threaten local fishermen’s livelihoods,” says Rosanna Marie Neil, a policy specialist with Northwest Atlantic Marine Alliance, which also opposes the legislation. “Instead of supporting the corporate takeover of our oceans, lawmakers should safeguard the livelihoods of fishermen and coastal residents who are already struggling.”

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For a decade, the U.S and European Union have blocked the import of raw molluscan shellfish from each other. Now, FDA and the European Commission (EC) have come to an agreement on the first-ever food safety equivalence determination for molluscan shellfish, which includes oysters, clams, mussels, and scallops. The landmark deal is in effect for the U.S. and Spain and the Netherlands, and opens up shellfish trade between the countries for the first time since 2010.

“These actions reflect key strategic international engagement and several years of careful review by the FDA on behalf of consumers at home and abroad,” Anna Abram, FDA’s deputy commissioner for policy, legislation, and international affairs, said in a prepared statement. “Today we’re helping unlock economic opportunity by creating a path forward to new market access for U.S. exporters.”

This deal was a long time in the making. FDA initially stopped allowing raw shellfish to be imported from EU countries in the late 1980s due to public health concerns. In 2010, the EU stopped accepting U.S. exports of raw bivalve molluscan shellfish after an EC audit determined that U.S. food safety controls did not comply with comparable EU requirements.

FDA says that experts on both sides of the Atlantic have spent the last number of years evaluating each other’s food safety control measures for molluscan shellfish and conducting on-site audits to verify the other’s systems. The two parties used food safety controls for molluscan shellfish with an approach laid out in the Veterinary Equivalency Agreement signed by both back in 1999.

Frank Yiannas, FDA’s deputy commissioner for food policy and response, tells Food Quality & Safety recently that a deal like this was expected, thanks in part to expanded traceability initiatives enacted as part of its New Era of Smarter Food Safety Blueprint. “Americans can be confident in the equivalence determination that Spain and the Netherlands have implemented safety controls that are equivalent to ours, thereby enabling us to allow Spain and the Netherlands to export raw molluscan shellfish to the U.S.,” Yiannas says. “The FDA is committed to keeping consumers safe and ensuring the safety of our food supply, and that includes seafood, whether it is imported or harvested domestically.”

Additionally, the EC will now permit raw and processed molluscan shellfish, including clams, mussels, oysters, and scallops, to be imported from the U.S., starting with shellfish from Massachusetts and Washington.

FDA plans to recognize other EU member states in the future, as arrangements have been made to use a streamlined process for expanding market access between these two trading partners.

Shellfish accounts for more than $1.6 billion in total value of U.S. exports, and this deal for shellfish trade is expected to push that number up considerably in the years ahead.

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Vibrio spp. represents a serious threat to human health. Three species in particular are linked to gastrointestinal issues and can lead to infections and septicemia: V. cholerae (VC), V. parahaemolyticus (VP), and V. vulnificus (VV). These pathogens are most commonly found in raw or undercooked seafood such as fish, squid, oyster, and shrimp. V. cholerae is the main factor that causes cholera, which is an important public health problem worldwide.

VP was first identified as a cause of foodborne illness in Japan in 1950 when 272 individuals became ill and 20 died after the consumption of semidried juvenile sardines. VP causes three major syndromes of clinical illness: gastroenteritis, wound infections, and septicemia. The most common syndrome is gastroenteritis. Symptoms of this syndrome include diarrhea with abdominal cramps, nausea, vomiting, headache, and low-grade fever. Strains from this pathogen that are isolated from diarrheal patients produce either the thermostable direct hemolysin (TDH), the TDH-related hemolysin (TRH), or both, while hardly any isolates from the environment have these properties.

In 2009, a Vibrio outbreak in Singapore was associated with consumption of Indian rojak (a traditional salad of fruits, vegetables, and seafood). The Singapore Ministry of Health concluded its investigations into the food poisoning cases and identified VP, traced to the cross-contamination of rojak and raw seafood ingredients harboring the bacteria as the source of the outbreak. Laboratory investigation confirmed 13 of the cases to be positive for VP, including the first fatal case.

The risk of these pathogens may only be getting worse. Scientists warn that, because climate change causes an increase in sea surface temperatures and a rise in sea levels, VP and VV infections will become more common. This is because warmer, rising waters create an even more welcoming environment for the deadly pathogen. Subsequentially, it is especially crucial that methods to efficiently detect Vibrio are developed.

Testing for Vibrio

FDA’s Bacteriological Analytical Manual (BAM) (Chapter 9) and the International Organization for Standardization (ISO) 21872-1:2017 are the two standard methods widely used for the detection of Vibrio. While these are the standard, there are still many issues that arise with these methods.

Neither of these methods provides a good selective enrichment medium for Vibrio species. Instead, different formulations of alkaline peptone water (APW) have been used as the preferred enrichment for certain Vibrio targets or food matrices. Still, no single enrichment procedure for classical isolation, by plating or selective media, has been validated by FDA or the ISO for all three strains.

Finding a single enrichment procedure that works for all three different strains is an important challenge faced by seasoned microbiologists today. The preferred enrichment temperature for VC is 42°C, but the preferred temperatures for VP and VV differ at 35-37°C. Furthermore, some food matrices containing high background flora or inhibitory compounds, such as bacterial growth or polymerase chain reaction (PCR) inhibitors, might require alternative enrichment schemes. In addition, the duration of enrichment and plating efficiencies of presumptive isolates could affect classical confirmation, making them difficult. Overgrowth of competing organisms might occur if enrichment duration exceeds 20 hours. This makes it difficult to isolate Vibrio on selective agar plates. Thiosulfate citrate bile salts sucrose (TCBS) agar is widely used as the main selective agar for isolation of the three target species by both the FDA-BAM and ISO methods.

Because climate change causes an increase in sea surface temperatures and a rise in sea levels, V. parahaemolyticus and V. vulnificus infections will become more common. This is because warmer, rising waters create an even more welcoming environment for the deadly pathogens.

Cultural confirmation is also a challenge. Not all isolates of the target species exhibit the same growth properties. Different isolates of the same species have shown as much as two logs differences in plate counts on TCBS plating efficiencies. This difference could be attributed to factors such as boiling time or depth of the poured media. Another challenge is that the Vibrio species might be subject to a biological phenomenon known as “viable but non-culturable.” When in this state, the pathogen is not able to be detected by traditional culture methods but is able to cause infection. A third challenge is that there are several atypical isolates of the target species, specifically for VP. Because of this issue, molecular-based methods, such as DNA sequencing, PCR-based methods, or matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) are good alternatives because they can confirm atypical Vibrio results, ultimately improving accuracy.

Standard methods are also labor intensive and rely on microbiological/biochemical identification. For seafood processors and inspections, current methods require at least three to five days for results and subjective interpretation for the screening of negative samples.

Food testing laboratories in the seafood industry are in need of a fast and accurate method to reliably detect the three main Vibrio species. An easy-to-use and rapid method that can reliably report results would allow seafood to safely get to market faster.

Real-Time PCR Detection

The advantages of real-time PCR are highlighted when used for the detection of Vibrio because of the challenges outlined above, such as the background flora naturally present in seafood matrices and the enrichment protocol challenges.

Since its invention, real-time PCR technology has been greatly improved so that it is more stable, accurate, and rapid for specific applications. As the technology evolved, new chemistries were developed based on fluorescence detection.

This evolution allowed for real-time PCR kits to achieve a high level of specificity and sensitivity when detecting Vibrio. Each test well can be used to detect all three important strains of the pathogen at the same time, decreasing the time it takes to get a result. A PCR kit may be able to detect the pathogen in 94 samples in less than two hours, rather than the typical three to five days. Further, the workflow is often optimized to be simple and user friendly.

While real-time PCR methods often offer quicker turnaround times than many of the standard methods, they can be prone to false positives due to free DNA from dead cells found in the sample. Emerging PCR-based methods should address this limitation.

Recently, Bio-Rad Laboratories received AOAC validation for its iQ-Check Vibrio assay. The assay uses a single-step enrichment followed by real-time PCR for the multiplex detection of VC, VP, and VV. This method provides rapid qualitative detection and differentiates among all three strains in seafood products. The solution also has an optional Free DNA Removal Solution that can address ambiguity caused by dead cell DNA by removing free DNA in the sample with a simple non-toxic protocol, while the intact DNA in living cells remains unaffected.

This method was evaluated and approved by the AOAC Performance Tested Methods (PTM 032002) program. Results of the AOAC-PTM validation study demonstrated no differences between the iQ-Check Vibrio method and the U.S. FDA BAM Vibrio reference method. The assay and the Free DNA Removal Solution were validated for use with 125-gram test portions of cooked and raw shrimp, raw mussels, raw oysters, and raw tuna. The assay was approved for use with Bio-Rad Vibrio Enrichment Broth (after a seven-to-nine hour enrichment period) and alkaline phosphate water (after a six- to 18-hour enrichment period), giving the user flexibility to optimize the method to their lab workflow, while significantly cutting down the traditional three to five days it takes to get results with standard methods.

Rapid methods like this one can greatly aid in outbreak investigation and management of public health concerns. The ability to obtain results in a shorter amount of time, particularly when it comes to pathogens such as Vibrio species, can be critical in reducing the impact from a food safety event.

Pastori is an international product manager at Bio-Rad Laboratories. Reach him at frederic_pastori@bio-rad.com. Wang is a field marketing specialist at Bio-Rad Laboratories. Reach him at weijia_wang@bio-rad.com.

The post How to Quickly Detect <em>Vibrio</em> spp. in Seafood Using PCR Technology appeared first on Food Quality & Safety.

In 1984, a 34-year-old Hawaiian resident of Japanese origin experienced intermittent abdominal pain coupled with inexplicable hunger pangs. Her diagnosis was the first of its kind to be reported in the United States: a confirmed case of parasitic penetration of the stomach lining. The identified cause of the infection was a third stage Anisakis simplex larva. From the patient’s food consumption history, it was evident that raw seafood such as tuna, salmon, and poki were a predominant part of her diet. This case and others got public health and environmental experts alike thinking more about zoonotic diseases.

Zoonosis occurs when the infection causing agent is transmitted between two different species, usually from animals to humans. Like anisakiasis, most zoonotic diseases are often mistaken for food poisoning because of the strong similarities between symptoms. The four primary manifestations of zoonoses may be gastric, intestinal, ectopic, and even as allergies. It is remarkable how the son of a watchmaker, Felix Dujardin, initially paved the path for a better understanding of zoonotic diseases. He first described the infection in 1845 by naming the worm he found in dolphins as Anisakis.

Surge in the Marine Parasite Population

A recent study conducted by the University of Washington revealed an alarming increase in the population size of marine parasites. The study demonstrated that, over the span of 40 years, the population size experienced a 283-fold increase in parasites. There exists a common observation between the 1986 findings of the research team that were observing anisakid nematodes within Hawaii and the 2020 study conducted by the research group from the University of Washington; salmon was more likely to be infected by marine parasites. This finding does not rule out the susceptibility of other seafood species to parasitic infections.

Possible Rationale Behind the Increase in Marine Parasite Population

Researchers are unable to pinpoint a specific reason behind the parasites’ population explosion. Various speculations (that are worth pursuing) co-exist, such as climate change, the introduction of fertilizers from field runoffs, global policies that preserve marine mammals (thereby increasing their population), and an increase in testing frequency. Over the past decade, the focus has intensified on our global water footprint, which has resulted in more extensive and frequent marine testing methodologies.

Understanding Seafood Parasite Behavior Is Important

It is important to convert awareness into action. The data obtained from studying seafood parasites helps us to conduct a pulse check on the overall “health” of the marine ecosystem, specifically of the mammals. If marine parasites prefer one species over another, then parasite population size could also be used as an indicator of the growth or suppression of other marine species.

A Food Safety and Quality Perspective

Sushi, sashimi, and ceviche have and will continue to remain popular delicacies around the world. With effective food safety measures in place such as obtaining seafood from trusted (and approved) suppliers, maintaining temperature control from dispatch until service, monitoring the pH (where applicable), and ensuring only fresh stocks are utilized, consumer safety can be assured. In most positive cases of zoonoses linked to salmon, the fish was obtained and served chilled, versus frozen.

For sushi lovers out there, it is a great idea to slice the piece of fish at least in half and do a visual check if you can, before consuming it. Even though some chefs may be meticulous with their visual checks, it is not uncommon for an unsuspecting parasite or two to slip away from the well-trained eye.

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Nestle is launching a new plant-based tuna alternative in Switzerland this month ahead of a global rollout, hoping that consumers eating at home during the COVID-19 pandemic will stay eager to try new products.

Nestle has been investing in plant-based food to make its prepared dishes unit trendier and more appealing to consumers wishing to lower their meat intake. The new “Garden Gourmet” tuna made with pea protein will be available in glass jars in the chilled aisle of Swiss supermarkets and can be used in salads, sandwiches, and pizzas. Ready-to-eat sandwiches will also be sold in some stores, Nestle says.

Developed by Nestle’s Swiss research facilities, the tuna is the group’s first plant-based seafood product to hit the market. Soy-based burgers, mince meat, sausages, and chicken nuggets are already available.

Nestle said last month that increased at-home consumption during the COVID-19 pandemic boosted demand for its Garden Gourmet plant-based products in the first half of 2020. The group’s sales of plant-based meat alternatives reached around 200 million Swiss francs ($218.7 million) last year.

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