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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Foodborne viruses can be tough to prevent and mitigate. Some can’t be cultured, so they are difficult to analyze. Others aren’t affected even by strong disinfectants, so intervention is ineffective. In the past decade, an additional virus, hepatitis E, joined norovirus and hepatitis A as a top three concern for human food safety.

To tackle these challenging foodborne viruses that can cause serious human illnesses, the Food and Agriculture Organization of the United Nations and the World Health Organization (FAO/WHO) are holding a series of meetings focused on microbial risk. The first Joint FAO/WHO Meeting on Microbiological Risk Assessment (JEMRA) convened in September 2023 in Rome and focused on foodborne viruses of top concern for public health, analytical methods, and contamination indicators. The second meeting, which took place in February 2024 in Geneva, discussed prevention and intervention measures for these viruses. A third meeting is planned for later in 2024 and will focus on evaluating risk.

The final reports for the first two meetings are still in progress, with only summaries released so far. Experts involved in the meetings said significant advances have been made in the study of foodborne viruses; these have helped researchers understand the science of viral mitigation since the inaugural JEMRA meeting 16 years ago, a milestone event that was the first time the issue of viruses in foods was brought to international attention.

“We have improved norovirus surrogates and ways to study human norovirus, and we have better detection methods, like digital PCR,” says Kalmia Kniel, PhD, associate chair of the department of animal and food sciences at the University of Delaware in Newark. She adds that thermal treatments are often relied on to inactivate viruses, but there are promising non-thermal technologies being studied, including cold plasma, chlorine dioxide, and some chemical disinfectant combinations.

The Biggest Threats

Dr. Kniel chaired the 2023 meeting and was a member of the expert committee that reviewed recent scientific developments, data, and evidence associated with foodborne viruses. JEMRA will update and provide scientific advice to the Codex Committee on Food Hygiene, which requested the series of meetings. The Codex committee will use the information for its international recommendations and standards. The expert committee also considered trade implications of possible standards to ensure that food safety does not become a trade barrier.

In reviewing viruses associated with human foodborne illness, the expert committee identified human norovirus as the leading cause of viral foodborne illnesses, followed by hepatitis A and hepatitis E. The ranking considered the frequency of illness, the clinical severity of the disease, and the food most often linked to the virus; however, while hepatitis A and hepatitis E were ranked equally behind norovirus in terms of frequency, they were higher than norovirus in terms of clinical severity. The committee lacked sufficient data to rank other viruses, including rotavirus and sapovirus.

In terms of the foods most associated with the viruses as a potential public health threat, prepared food, frozen berries, and shellfish—in that order—are associated with norovirus. For hepatitis A, linked foods are shellfish, frozen berries, and prepared foods. Those two viruses are transmitted via contamination by feces exposure. For hepatitis E, a zoonotic virus, pork and wild game are associated, and the virus is transmitted from animal to human.

The committee considered only water used in food production, in processing, in preparation, or as an ingredient, not water intended only for drinking, in its assessments.

Viral foodborne disease has a substantial impact on morbidity and mortality globally, but surveillance data is sparse, and there is the potential for asymptomatic shedding, so it is difficult to craft prevention and control strategies.

Norovirus causes about 125 million cases of foodborne illness and 35,000 deaths worldwide annually, according to the committee’s summary, including severe outcomes such as hospitalization and death, especially in children younger than five years old, the elderly, and immunosuppressed people, who may shed the virus for extended periods. Hepatitis A causes about 14 million cases of foodborne illness and 28,000 deaths each year globally, but there are significant regional differences attributable to endemic prevalence, vaccine use, and international food trade. There are no global estimates for hepatitis E, which can damage the liver, the meeting summary said.

“The JEMRA committees discuss foodborne viruses in a global context,” Dr. Kniel said. “We need to keep in mind that our food system is global in nature, which means we need better surveillance of viruses in all countries in order to help each other.”

Dr. Kniel said that since the 2008 JEMRA report, international and national standard methods have been developed and validated to detect and quantify human norovirus and hepatitis A virus in foods. Methods released since that report include the International Standards Organization’s ISO-15216-1:2017 and ISO-15216-2-2019. These are used widely to detect norovirus and hepatitis A in leafy greens, soft fruits, and shellfish, and as a benchmark to validate new methods, the committee’s summary said. There is no ISO method for prepared foods. Methods to detect hepatitis E in meats are under development.

The committee said infectivity assays are needed for wild-type viruses, as there still is no definitive way to tell infectious from noninfectious viruses using molecular amplification.

It recommended that countries consider building capacity to help with adopting and training in methods for detecting viruses in foods and the environment. “Appropriate global actions will help alleviate the anticipated increase in public health risk from viral foodborne illness arising from population growth, the climate crisis, and globalization of food supply chains,” the summary from the 2023 meeting said.

Prevention and Mitigation

Prevention is the preferred focus now, given the difficulty and expense of mitigating infected foods, says Lee-Ann Jaykus, PhD, rapporteur of the March 2024 meeting and a member of its expert committee. She says the viruses are not culturable organisms and cannot be grown in a lab like bacteria can, nor can they be culturally enriched. There is no host cell in a culture with which to propagate them. It’s necessary to concentrate and purify them from a sample and use reverse transcription polymerase chain reaction to detect the viruses. “We have standardized methods to detect these viruses in selected commodities, but they have some inherent disadvantages because of the limitations of not having a culture,” Dr. Jaykus says.

Limitations include the fact that even when a viral nucleic acid is detected, it doesn’t necessarily mean there is an infectious virus, she said. Real-time polymer chain reaction (RT-PCR) is a complicated method, and it is easy to lose viruses in the first steps, so it is not as sensitive as needed. “These are limitations not because the science is bad,” she says. “The science is the best it can be as it currently stands. There are limitations because we can’t grow these things.”

One focus of the second meeting was contamination routes for the virus to humans. Fecal matter and vomit from infected humans are the primary sources of contamination for norovirus and hepatitis A to get to humans through affected waters, food handlers carrying the viruses, and surfaces, because the viruses can live for weeks on surfaces, Dr. Jaykus says. The zoonotic hepatitis E virus is present in the meat, organs, tissues, and excretions of infected swine and game animals and gets transmitted through exposure.

Because the viruses persist in the environment for long periods and are resistant to many treatments, prevention is the key strategy to control foodborne viruses, Dr. Jaykus says. One example of prevention is reducing the viral load in shellfish by treating wastewater, but that requires infrastructure investment. Another is using production-related strategies to reduce contamination of fresh and frozen produce. Virus inactivation methods also are under investigation.

The committee recommended some directions for future research and development, including early identification of contamination hotspots using wastewater surveillance, for example, and technologies such as satellite imagery and hydrographic dye studies to predict virus dispersion. It also recommended using emerging scientific data to develop surface disinfectants and hand sanitizer formulations with greater efficacy against environmentally stable viruses. After all, hand sanitizers were effective in reducing transmission during the COVID-19 pandemic.

“Following up on the COVID-19 pandemic, it is critical that we launch surveillance studying the health of animals, humans, and the environment to identify important zoonotic viruses before the next pandemic,” says Dr. Kniel, who, like Dr. Jaykus, was surprised to see the hepatitis E virus added to the list of top foodborne virus concerns since the inaugural JEMRA meeting 16 years ago. “It is frustrating to continually talk about the need for better surveillance to better understand foodborne virus transmission and the attribution of disease to a specific virus.”

Table 1: Foodborne viruses and foods of highest public health concern

Source: FAO/WHO.

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Safety is the foundation of food quality. It’s critical that any laboratory, manufacturing plant, and production facility that handles food maintains the highest standard of safety and follows careful procedures to the letter. There’s no room for error; a single isolated shortcut can lead to disastrous results.

The USDA estimates that one in six Americans become sick from foodborne illness each year. Since New Year’s Eve 2023 alone, the agency’s Food Safety and Inspection Service (FSIS) site’s Recalls and Public Health Alerts page has featured multiple cross-contamination incidents. The alerts warn consumers away from specific food products, with concerns such as “possible Salmonella contamination,” “possible E. coli contamination,” and “possible extraneous material contamination.”

Here are five safety steps that are absolute musts when it comes to preventing cross contamination and running a squeaky clean and safe laboratory.

1. Institute, Document, and Mandate Cleaning Procedures and Techniques

Everyone who steps foot in a lab space should be educated about agreed-upon safety procedures so they can be consistently followed. Methods must to be adhered to without fail; something that was “barely used” or “looks clean” is not acceptable.

• Prevent cross contamination by moving from high to low in a cleaning cycle, such as cleaning shelves above a workspace before the workspace itself.
• Avoid cleaning while testing is taking place.
• Keep equipment clean at all times, without exception. This means wiping down equipment after every use and scheduling regular deep cleaning as applicable.
• Establish and follow a cleaning checklist to prevent the risk of a missed or over-looked step. Checklists can also help keep others in the lab informed so there is no miscommunication.
• Know the risks of cross contamination and institute fail-safe cleaning methods. For example, pipettes are a leading cause of cross contamination within a lab setting. Best practice in equipment sanitation is to completely sterilize, not just clean, if possible. Sterilization can include disassembly and autoclaving for at least 20 minutes at 121ºC (252ºF). Each lab should have the procedures for the type of use and equipment outlined in detail.

2. Maintain Proper Air Circulation and Ventilation

Surfaces are not the only source of contaminants; the air within a closed room can harm employees and contaminate food or samples. Air handling in a food lab is not the same as air handling in a non–food-related commercial operation.

• Air handling in a food lab begins with a risk assessment to identify the unique risks within the building. For example, establishing positive air pressure zones is an important aspect of air flow design in a lab, but older buildings tend to have multiple exhaust fans, and exhaust fans create negative pressure zones.
• Hygienic design of air handling units (AHUs) and ducts is imperative to food safety. Employ an HVAC engineer to design a system with the appropriate number of air turns per hour to fit the facility and its operations.
• Standard ventilation filters could be blowing contaminants in the lab. The level of food micro-sensitivity will dictate the level of filter standards and the type of filter needed.
• Air sampling can help to determine if the air within a lab space has high levels of microbiological activity.

3. Maintain a Tidy Workspace

While this may sound as if it goes without saying, workspaces that aren’t carefully cleaned can harbor microorganisms, bacteria, and allergens. This can endanger employees within the lab and increase the risk of cross contamination.

• Labs should be organized so that expectations are crystal clear. A good rule to follow is the “5-S” process: sort, set in order, shine, standardize, and sustain.
• Dispose of expired products promptly and ensure that they don’t come into contact with lab equipment or samples.
• Use only designated cleaning tools, solutions, and products, and create timetables to regularly switch them out.
• Clean the lab area in the moment and/or at regular intervals throughout the day (whichever comes first).
• Mandate and provide gloves and other personal protective equipment for all personnel to protect against cross contamination and contain lab testing within smaller areas.
• Utilize designated disposal bins for different testing waste, keeping biohazard waste and chemicals separate from non-biohazard waste.
• Design storage with safety in mind. Designated safety cabinets help workspaces stay organized, but they can also increase safety levels in a lab.

4. Keep Equipment in Pristine Condition

Faulty equipment escalates multiple risk factors—namely, biohazard risk, food safety risk, and personal safety risk. Even if equipment is perfectly clean, it can leak or create other messes that contribute to an unsafe lab if it’s not in top condition.

• Document and communicate regular maintenance and inspection schedules. Equipment needs to be regularly tested and proactively inspected to verify its condition.
• Implement a robust system of checks and balances to ensure that maintenance activities are not isolated.
• Clean equipment according to the operator’s manual. For example, distilled water and specialized cleaning agents may be required to keep equipment operational and prevent corrosion.
• Keep equipment free of dust, dirt, grease, and of course bacteria to improve performance and increase safety.
• Focus on preventative maintenance to extend the life of equipment productivity.

5. Test, Test, and Re-Test Within Your Lab Setting

Even the cleanest facilities need to ensure that their cleaning procedures are effective.

• Perform regular environmental testing to check the lab environment.
• An environmental monitoring program (EMP) can determine whether or not an environment is sanitary and verify if pathogen controls are working.
• Utilize negative control plates when using microbiological samples to check for cross contamination.
• Enlist food safety partners to assess tests within the lab (additional checks/balances).

The Cleaning Supply Chain

As in the distribution supply chain, one weak link in a laboratory can affect the entire chain. A lab may have extensive protocols in place to keep equipment clean and fully operable, but if a new employee is unfamiliar with the equipment, the process can start to break down.

Training at All Levels

Food labs and production facilities can amp up the level of safety by seeking supply chain partners that offer training on the equipment they provide, including usage, maintenance, and cleaning. At the other end of the chain, a food distributor, wholesaler, or retailer needs training to continue the chain of safety.

A Culture of Safety

Another important aspect to be aware of in a supply chain partner is company culture. While this can be harder to discern at first glance, there are red flags that can indicate that an organization’s values may inadvertently affect the level of safety. For example, a focus on speed over all else may lead to shortcuts or hasty cleaning protocols that increase safety risk all the way across the chain.

Consumer Safety

Both contract and in-house labs help prevent contamination and foodborne illnesses. As time goes on, their role in analyzing and mediating safety issues at a larger scale is increasing. In other words, food labs are equipped with the tools and expertise to perform analytical and preventive work that can support the entire food system, not just its direct partners.

A Look to the Future

Labs that are efficient, productive, and clean enable vendors and suppliers to provide safe food to the growing populations of consumers across the globe. Impeccable cleaning protocols can protect public safety and also allow food companies to channel resources toward growth initiatives, rather than using those same resources to cover the damaging expenses of recalls. With safety as a foundation, food labs play a central role in the future of our food systems.

Kotecki is a technical sales manager for Nelson-Jameson. Reach her at k.kotecki@nelsonjameson.com.

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Imagine that disaster strikes on the production line just weeks before a candy company’s famous treats flood grocery store shelves. Poor process controlled to crystallized sugar being added during a stage that destroyed the batch. When alarms sounded hours later, thousands of candy boxes had already been filled with grainy goods. With their signature smoothness missing, the brand’s subpar candies became an embarrassing and costly mistake. Lacking control of operational processes can hinder product excellence and negatively affect brand reputation; however, inline, accurate, repeatable process monitoring can prevent quality and safety catastrophes from the earliest stages of transforming raw materials to putting on the finishing touches.

When every sugar crystal, drop of syrup, and piece of candy demands accurate process monitoring, the art of cane and beet sugar processing, syrup production and confectionery production meets the science of measurement accuracy.

Precise Control from End to End

Every process involved in sweet treat production requires careful process control and measurement accuracy. Accurate, repeatable measurement of raw and in-process liquids at multiple processing steps empowers producers and processors to optimize disparate processes for maximum quality and efficiency within facilities.

Creating sugar from raw materials. Processing harvested sugarcane and sugar beets creates edible sugar. After it is extracted, the juice or syrup is purified to remove any contaminants or solid particles. Heating the clarified liquid helps evaporate excess water, leaving behind a concentrated blend of flavors and sugars. From there, sugar crystals form and separate from the remaining liquid after the liquid sugar solution cools through manipulation of temperature, humidity, and movement.

Strict processing parameter adherence prevents process and product quality deviations. Monitoring Brix levels during raw materials processing ensures an accurate seeding point and optimal crystallization. Inline refractometers continuously monitor the liquid concentration to carefully control the cooked solution and crystallization process. Going beyond the target Brix level risks crystal conglomeration, which can result in wasted batches and costly reprocessing.

Syrups preparation. Syrup producers then blend the concentrated sugar solutions with various flavors and other ingredients to create syrups varying in taste, texture, and appearance. Melters utilize elevated temperatures to achieve the desired solubility, viscosity, and chemistry. Accurate, reliable liquid concentration measurements minimize cooking time and ensure even blending. Even better, the ability to precisely control Brix levels supports the uniformity of the syrup mixture, guaranteeing consistency in taste, texture, and appearance. Confectionery manufacturers then buy the ready-made sugar or syrups to develop their products.

Confectionery manufacturing. Sugar confectionery and chocolate filling makers purchase processed sugars or syrups to create their products. These sugary treats include candies, chocolate fillings, chewing gum, marshmallows, and other desserts rich in sugar and carbohydrates. To maintain a specific shape, texture, flavor, consistency, and overall quality, the sugar content in the products must be concentrated at a desired level through cooking and evaporation of water.

Inline process refractometers provide continuous, real-time information throughout the pipeline to help determine the end point and ensure consistent product quality. Ideal for confectionery manufacturing and candy filling manufacturing machinery, these retractable instruments eliminate the need for sampling, prevent process disruptions, and save valuable processing time; however, measuring liquid concentration and Brix during different applications to craft the perfect sweet treat comes with its share of difficulties. 

Inline, Continuous Measurement

Temperature, natural variations in the raw material-filled juice concentration and the sugar content of syrups and other confectioneries, and additional factors can impact the final product quality, thereby affecting customer satisfaction.

First, sugar processing involves multiple stages with varying temperature requirements exceeding 150°C. From melting to evaporation to refining, each process involves incredibly high heat, so refractometers must endure temperatures up to 150°C, or 300°F, for accurate functionality.

Figure 1. The final Brix of the mixture determines the flavor, consistency, and overall quality of the final product. Courtesy of Vaisala.

Sugar syrups, confections, and candy fillings often involve intricate mixtures of various ingredients, each contributing to the end product’s overall flavor profile, appearance, consistency, and more. Measuring the liquid concentration of syrups, sugar confections and chocolate fillings throughout blending and mixing processes can be difficult—and unreliable—with a handheld refractometer and manual sampling dependent on human error.

With inline process refractometers reliably measuring the liquid phase and Brix across each process in real time, from processing to syrup prep to confectionery and filling production, decision makers can realize numerous advantages and produce top-grade products.

Liquid Concentration

While sugar processing, syrup preparation and candy-making processes all pose technical hurdles, the benefits of proper liquid concentration and Brix measurement benefits are substantial.

Improved product quality and consistency. Accurate measurements drive the creation of confections with unparalleled quality and consistency. Confection makers can create products that reliably meet or exceed consumer expectations by ensuring consistency across batches. Preventing under- or overconcentration eliminates flavor disruptions, strange textures, or variable melt points.

Substantial cost savings. Without accurate Brix data, manufacturers might struggle to stay within recommended material levels, quickly running through resources to adjust the sweetness or flavor. Fine-tuning various processes based on accurate liquid concentration and Brix measurements enables manufacturers to reduce waste and extract the maximum value from their materials. Optimized process control translates into sizable cost and ingredient savings.

Fewer labor-intensive tasks. Bet­ween manually monitoring the crystallization, evaporation, extraction, blending, mixing and other confectionery production processes, as well as taking samples and making adjustments in the event of deviations, each step from raw material processing to confirming the final product quality can be arduous and time-consuming. Automated inline measurements reduce the need to collect samples, run tests, and control processes manually to account for divergence.

Peace of mind. Overall, Brix gives confectioners a window into multiple process parameters beyond enhancing quality and efficiency. Reliable inline Brix measurement also provides invaluable peace of mind. Avoiding intermittent manual sampling eliminates risks associated with human error and contamination. Tight instrumentation regulation, within limits, ensures that any variations or deviations are caught instantly before they impact the end product. Additionally, by avoiding taking manual samples from product during heating or melting, technicians are no longer exposed to burns. Germs and particles introduced through manual sampling are also avoided.

As sugar mills and refineries, syrup producers, and various confectionery and filling makers depend on the reliability of refractometry for liquid concentration measurements in myriad applications, advanced continuous, inline measurement and monitoring systems maximize production efficiency, product quality, and profits by decreasing discarded ingredients, failed batches, and repetitive manual steps.

Harness Innovation

Countless confectioners face catastrophic quality failures and losses stemming from poor process and quality control. Accuracy, repeatability, and safety unlocked by inline measurements are the keys to perfection in the world of candies and sweets.

Off-target concentration levels critically impact product quality, flavors, melt properties, and more. Inline process refractometers equip sugar processors, syrup preparers, and confection makers with a powerful tool to elevate their products, providing real-time, continuous monitoring of liquid concentration and allowing for instantaneous adjustments.

With inline refractometry, decision makers finally eradicate the crippling product quality issues and profit shortcomings rooted in poor concentration control. Precisely measuring edible sugar, sugar syrups, and confectionery creations at every stage translates directly into improved product quality and safety, reduced costs, and enhanced process control, all of which contributes to consumer delight, bite after bite.

Green is a regional sales manager at Vaisala, a provider of industrial measurement and instrumentation solutions for the food, beverage, and agriculture industries. He has a background in mechanical engineering and brings experience in instrumentation, sales, and product management. At Vaisala, he is primarily focused on measurement applications applied to food and beverage production.

Figure 1. The final Brix of the mixture determines the flavor, consistency, and overall quality of the final product.

Courtesy of Vaisala.

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Consumer Reports (CR) has called on USDA to remove certain Lunchables food kits from the National School Lunch Program due to high levels of sodium and heavy metals found in the products.

USDA currently allows two Lunchables kits—Turkey & Cheddar Cracker Stackers and Extra Cheesy Pizza—to be served to nearly 30 million children through the National School Lunch Program. To meet the program’s requirements, Kraft Heinz added more whole grains to the crackers and more protein to the Lunchable kits designed for schools, compared to store-bought versions.

CR recently compared the nutritional profiles of two Lunchable kits served in schools and found they have even higher levels of sodium than the kits consumers can buy in the store. CR also tested 12 store-bought versions of Lunchables and similar kits and found several contained relatively high levels of lead and cadmium. All but one also tested positive for phthalates, chemicals found in plastic that have been linked to reproductive problems, diabetes, and certain cancers.

CR tested store-bought Lunchables and similar kits from Armour LunchMakers, Good & Gather, Greenfield Natural Meat Co., and Oscar Mayer and found lead, cadmium, or both in all. Lead and cadmium can cause developmental problems in children over time, even in small amounts. While none of the kits exceeded any federal limit, five of the 12 tested products would expose someone to 50 percent or more of California’s maximum allowable level for lead or cadmium – currently the most protective standard.

The sodium levels in the store-bought lunch and snack kits CR tested ranged from 460 to 740 milligrams per serving, which is nearly a quarter to half of a child’s daily recommended limit for sodium. CR found that the sodium levels of the Lunchables made for schools, which had a larger portion of meat, are higher than in the store-bought versions. The school version of the Turkey and Cheddar Lunchable for schools contained 930 mg of sodium compared to 740 mg in the store-bought version. Similarly, the Lunchable pizza kit for schools had 700 mg of sodium compared to 510 mg in the store version.

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On April 10, the Environmental Protection Agency (EPA) implemented first-ever restrictions on the perfluoroalkyl and polyfluoroalkyl (PFAS) substances in drinking water, a pivotal move in shielding public well-being from waterborne hazards.

EPA’s cap target six PFAS compounds, including two of the oldest and most widespread PFAS—PFOA and PFOS—at 4 parts per trillion. The rule also sets limits of 10 ppt for PFHxS, PFNA, and HFPO-DA (also known as GenX), thereby establishing a benchmark for the most stringent health thresholds concerning these impurities in potable water.

Under the new rule, public water systems are required to monitor these PFAS compounds, with an initial monitoring period of three years, concluding by 2027, followed by ongoing compliance checks. Additionally, these systems must disclose information regarding the levels of these PFAS in drinking water, commencing in 2027. Further, public water systems are allotted five years—until 2029—to implement remedies aimed at decreasing PFAS levels if monitoring reveals that these levels exceed the designated maximum contaminant levels (MCLs).

Nicknamed “forever chemicals” because of their resistance to be degraded or destroyed, PFAS have been associated with several health issues, including high cholesterol, cancer, and thyroid disease. “There’s no doubt that these chemicals have been important for certain industries and consumer uses, but there’s also no doubt that many of these chemicals can be harmful to our health and our environment,” Michael Regan, EPA Administrator, said on a call to media this week.

Starting in 2029, public water systems found to have PFAS concentrations in drinking water surpassing the MCLs must take measures to reduce these levels and notify the public of the violation.

In an effort to help with enforcement, EPA announced it would make $1 billion in funding available through the Bipartisan Infrastructure Law to help states implement PFAS testing and treatment at public water systems and to help owners of private wells address PFAS contamination.

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Detecting foodborne bacteria has never been easy. Pathogenic bacteria are often sparsely distributed throughout food batches and can be present in very low numbers in randomly collected food samples. Further, the diversity of normal flora found in various food matrices creates a complex and dynamic microbial ecosystem that could interfere with the detection of target pathogens. Microbial dynamics are also influenced by food’s microstructure and chemical composition, which adds additional complexity to the detection process. Consequently, the detection of foodborne pathogens requires, first and foremost, accurate and reliable techniques to effectively maintain food safety.

Conventional culture-based methods, which were developed and implemented several decades ago, continue to deliver a reliable but conservative solution, capable of detecting as few as one target cell per 25g to 325g food sample. These methods are still widely regarded as gold standards for detecting foodborne pathogens, due to their precision and accuracy; however, these traditional methodologies are time-consuming, taking more than a week to provide a final result. Moreover, each identified pathogen requires independent protocols, which is neither convenient nor compatible with today’s intense production needs.

To address the shortcomings of traditional detection methods, numerous molecular techniques have been developed and used effectively in the past decade to detect foodborne pathogens. The development of real-time quantitative polymerase chain reaction (qPCR) has revolutionized microbiological analysis by enabling the detection of pathogenic microorganisms in food without the need for the labor-intensive and time-consuming procedures of isolation and identification. This method has dramatically reduced the time-to-result, which is a critical performance standard used to evaluate the effectiveness of a detection tool, alongside assay sensitivity.

The qPCR method not only provides faster, more sensitive, and specific results than traditional PCR methods, but also offers the potential for multiplexing, which means it can simultaneously detect multiple pathogens in the same reaction, enhancing operational efficiency and reducing overall costs. Numerous food commodities, including shellfish, fresh fruits and vegetables, dairy, and meat products, have been found to be contaminated with multiple pathogens of concern, such as Salmonella enterica, Listeria monocytogenes, and Escherichia coli, along with diverse species of Shigella, Campylobacter, and Vibrio. Consequently, simultaneous detection of multiple pathogens on a single-assay platform aligns with contemporary food industry trends and could also mitigate industry and regulatory needs in the mandatory testing of food products for a range of pathogens prior to distribution.

The Enrichment Step

While the potential advantages of qPCR multiplexing may seem apparent, it’s important to consider the sensitivity of these detection platforms. To guarantee the achievement of legal limits (absence in 25g for most bacterial pathogens), an enrichment step using microbiological culture media is still required prior to qPCR detection. Integrating both traditional microbiological enrichment and molecular pathogen detection serves as a useful bridge that links traditional and molecular microbiology. This approach offers combined benefits while also reducing some of the limitations associated with each method.

If performed appropriately, a short enrichment step is typically sufficient to “produce enough DNA” for subsequent qPCR detection. Moreover, the enrichment process not only increases the concentration of target pathogens in the sample but also revitalizes physiologically stressed or injured microbial cells. Selective enrichment is also crucial for suppressing the naturally occurring background microorganisms, enhancing detection efficiency, and preventing false positive outcomes; however, some of the drawbacks of selective enrichment media include the inhibitory nature of selective agents, which may slow down the growth or even suppress recovery of healthy or stressed target pathogens, ultimately impacting the detection process.

Numerous microbiological culture media with optimized selectivity have been validated and commercialized for short, single-step enrichments for the detection of foodborne pathogens such as Salmonella, Listeria, E. coli, and Campylobacter, across a variety of simplex qPCR assay platforms.

Multiplex qPCR Assays

When a multiplex format is desired, the situation becomes significantly different. Most multiplex qPCR applications are non-commercial and developed in house and open assays, which require standardization and quality assurance for molecular diagnostics. Additionally, multiplex diagnostics are only effective at detecting all target pathogens if they are properly enriched to detectable levels. Overcoming this challenge is difficult since the optimal conditions for detecting one pathogen may not benefit another, and competition among microflora can negatively affect the detection of other pathogens.

Currently, two different approaches are used to enrich food samples prior to detection by multiplex qPCR assays. The first approach involves using non-selective media, such as buffered peptone water (BPW) and universal pre-enrichment broth (UPB), for simultaneous enrichment of multiple foodborne pathogens, including Listeria, Salmonella, and E. coli, in food and environmental samples, followed by detection using multiplex qPCR. However, these broths may recover and enrich target pathogens along with background flora, which can lead to false negative detection results, particularly when complex interfering flora is present in the tested food samples. Therefore, using traditional non-selective enrichment media may not be appropriate for samples with high levels of background microflora, such as raw or unprocessed samples from animal and plant origins.

The second approach employs traditional selective enrichment broths, which help to eliminate interference from background flora in food samples; however, this method necessitates separate enrichment for each type of bacteria being tested. Once each bacterium is individually enriched, small aliquots of the samples are combined into a single diagnostic run. Nevertheless, as multiplex detection platforms evolve to handle several pathogens in a single assay format, it is also important that the enrichment procedures evolve accordingly. Ideally, a single enrichment medium should be used to fully take advantage of multiplexing capabilities.

Desirable for multiplex detection, a multi-pathogen enrichment broth should have the capability to recover sublethally injured cells and selectively enrich all target pathogens from complex background flora in each single or composite food sample. The development of a universal multi-pathogen enrichment medium has become urgent for enabling the simultaneous recovery and concurrent growth of multiple types of bacteria in a single step, making multiplex testing more efficient and cost effective.

Media Development

Developing universal multi-pathogen media requires the careful consideration of several crucial features to guarantee optimal effectiveness and functionality. One such essential characteristic is the ability to simplify the testing process by reducing the required enrichments. Hence, the media should be designed to support multiple bacterial types in a single broth, allowing for greater versatility in testing various food commodities for different target bacteria. This flexibility can streamline the workflow in a food safety testing lab by eliminating the need for multiple culture media preparations and minimizing the risk of errors. Consequently, multi-pathogen media should be formulated with specific nutrients and inhibitors to promote the growth of target bacteria while inhibiting the growth of other bacteria. Such selective enrichment increases the concentration of target bacteria in the sample, improving the sensitivity of subsequent multiplex detection assays, thus ensuring more reliable and accurate outcomes and reducing the risk of false positives or false negatives.

Additionally, multi-pathogen media can enhance the recovery of stressed or sub-lethally injured bacteria that may not grow well in traditional enrichment media, thereby eliminating the possibility of false negative results. The ability to detect stressed or injured pathogenic bacteria is highly desirable in the safety testing of various processed food commodities, such as pasteurized dairy products, deli meats, canned food products, and others. This is because the presence of such bacteria can pose significant risks to consumer health. By identifying these harmful microorganisms, appropriate measures can be taken to prevent their proliferation and minimize the chances of foodborne illness.

Another notable feature of multi-pathogen enrichment media is their ability to support the concurrent growth of different types of bacteria with varying nutritional requirements. Traditional enrichment media are often formulated with specific nutrients to support the growth of a particular type of bacteria, which may not be suitable for other types of bacteria. In contrast, multi-pathogen enrichment media must be formulated with a broad range of nutrients that can simultaneously support the growth of different types of bacteria. Adding specific repair-stimulating and growth-boosting factors, such as siderophores, amino acids, phospholipids, vitamins, and minerals, can help improve recovery and reduce adaptation period for slow-growing bacteria, making multi-pathogen enrichment media versatile and adaptable for various food safety testing applications.

Moreover, multi-pathogen enrichment media can be tailored to specific food matrices, which may vary in their composition and characteristics. Different food types, such as meat, poultry, dairy, fresh produce, and processed foods, may require different enrichment media to facilitate effective recovery of target bacteria due to variations in pH and osmolarity, nutrient content or preservatives, etc. Multi-pathogen enrichment media that contain buffering systems, and osmoprotective and neutralizing molecules can provide optimal conditions for the growth and recovery of specific pathogens in different food matrices. This can result in more accurate and reliable food safety testing outcomes, ultimately enhancing the safety and quality of food products.

Recent investigations into multi-pathogen enrichment media for multiplex foodborne pathogen testing have shown great promise in developing a universal selective enrichment broth. By balancing concentrations of different selective agents and optimizing selectivity levels, it is now possible to achieve simultaneous enrichment of some of the most prevalent foodborne pathogens, such as L. monocytogenes, Salmonella, Shigella, E. coli, and Staphylococcus aureus. While these media formulations have primarily been tested using in-house multiplex qPCR detection platforms, they are not yet commercially available.

As an alternative, in the case of difficult-to-culture foodborne pathogens such as Campylobacter spp., which have unique growth requirements and cannot be enriched in a universal multi-pathogen medium, multiplex testing can be achieved through the combination of enrichment aliquots into a single multiplex run sample.

Multiple Benefits

Using multi-pathogen enrichment media not only provides optimal growth conditions for specific pathogens in different food matrices, but also reduces the cost and storage space required to manage them in a food safety testing lab. This is particularly beneficial for labs with limited resources or space constraints. In addition, multi-pathogen enrichment media can also contribute to sustainability efforts in food safety testing labs. Traditional enrichment media often generate a significant amount of waste, including leftover liquid media and disposable plastic containers. In contrast, using multi-pathogen enrichment media can reduce waste by requiring fewer media formulations and packaging materials. This can promote an environmentally friendly approach to food safety testing, aligning with the increasing focus on sustainability and eco-friendly practices in the food industry.

While multi-pathogen enrichment media offer numerous benefits and features, they require careful validation to ensure compatibility with different multiplex detection platforms. Each multi-pathogen enrichment medium must be validated for its ability to effectively enrich the target bacteria and meet the sensitivity requirements of multiplex assays in detecting these bacteria. This validation process ensures that multi-pathogen enrichment media are reliable and accurate for use in food safety testing, enabling the detection of multiple pathogens in a single assay.

Incorporating universal multi-pathogen media into the enrichment process before testing complements the multiplex detection platform as a comprehensive package technology that evolves to handle multiple pathogens in a rapid, single assay format. The use of the media is a valuable tool in food safety testing, providing numerous benefits and features that can enhance the efficiency, accuracy, and sustainability of the testing process. As food safety remains a top priority in the food industry, the adoption of multi-pathogen enrichment media can significantly contribute to more efficient and reliable multiplex detection methods, ultimately protecting consumer health and well-being.

Dr. Olishevskyy is vice president of research and development FoodChek Laboratories in Sainte-Julie, Quebec, Canada. Reach him at solishevskyy@foodcheksystems.com.

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This week, FDA issued an import alert for human food products with detectable levels of chemical contaminants that may present a safety concern to human health. The Import Alert 99-48, Detention without Physical Examination of Foods Due to Chemical Contamination, gives the agency the ability to help prevent entry of human food products into the U.S. if they are found to be contaminated with a broad range of human-made chemicals including benzene, dioxins and polychlorinated biphenyls (PCBs), and per- and polyfluoroalkyl substances (PFAS), among others.

PFAS are a diverse group of thousands of chemicals used in many different types of products. PFAS in the environment can enter the food supply through plants and animals grown, raised, or processed in contaminated areas. It is also possible for very small amounts of certain PFAS to enter foods through food packaging, processing, and cookware.

In 2022, FDA initiated a targeted survey for PFAS in 81 seafood samples collected at retail and determined that the estimated exposure to perfluorooctanoic acid (PFOA), a type of PFAS, from certain samples of canned clams from China is likely a health concern. The 81 samples in the survey consisted of clams, cod, crab, pollock, salmon, shrimp, tuna, and tilapia, most of which were imported to the U.S. The agency plans an additional targeted survey of molluscan shellfish this year, and this new import alert could be used to refuse entry of foods such as seafood contaminated with PFAS.

Specific firms and their food products found with levels of chemical contaminants that may pose a risk to human health may be subject to detention without physical examination under the new alert.

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A group of food scientists from Cornell University in New York have concluded that a standard quality test often used for raw organic milk is not sufficient for differentiating between specific groups of bacteria and should be updated. The research appeared in January in the Journal of Dairy Science (doi: 10.3168/jds.2023-24330).

Nicole Martin, PhD, senior author of the study and assistant research professor in dairy foods microbiology and the associate director of the Milk Quality Improvement Program in the Department of Food Science at the institution, says that the present test, known as the laboratory pasteurization count (LPC), searches for thermoduric bacteria, but doesn’t distinguish whether bacteria form spores or not.

She notes that when dairies deliver organic milk to processors, the milk is sometimes tested for thermoduric bacteria using LPC. Under current standards, if thermoduric bacterial counts are high, the milk can be downgraded or even rejected by the processor. “We saw firsthand the struggle that some dairy farmers had with controlling and troubleshooting the LPC in their raw milk, which selects for thermoduric bacteria, or bacteria that can survive temperatures considerably above their maximum growth temperature,” Dr. Martin tells Food Quality & Safety. “This can lead to loss of premiums and, ultimately, even loss of contracts if the LPC is not brought back into compliance.”

With that in mind, the researchers noted that the criteria for determining milk quality at processing plants is no longer valid and a new way for producers to address milk-production hygiene is necessary.

The researchers went into the study looking to answer a few questions about the LPC that they hear frequently from farmers and other stakeholders, including, “Can milk be frozen prior to LPC testing?” “What are the types of organisms making up the thermoduric population in organic raw milk?” and “Can a rapid identification method used primarily for mastitis organisms be used to identify thermoduric bacteria?”

“So, it’s not necessarily that the LPC is insufficient, but that the LPC alone can only give us so much information, and for farmers who are actively trying to reduce LPC, it may be beneficial to understand the types of bacteria leading to the elevated LPC,” Dr. Martin adds. “This would also for more targeted troubleshooting efforts.”

The research showed that there are two different groups of bacteria making up the thermoduric population in organic raw milk—sporeformers and non-sporeformers—and an individual milk sample with a high LPC may have one or the other, or both, of these types. “Once we know what type of bacteria is driving the elevated LPC, it then allows for more targeted troubleshooting since these groups of bacteria are likely to originate from different sources on the farm,” she says. “We’re giving organic farmers the knowledge they need to make high-quality raw milk and for it to be economically viable; it will make a better dairy product in the end.”

The investigators concluded that, when troubleshooting elevated LPC, it is beneficial to know what the predominant type of thermoduric bacteria are that are contributing to the LPC. “Overall, our research shows that organic raw milk quality is very good, but some producers occasionally deal with high bacteria levels, and often it can take a lot of time and resources to resolve those issues,” Dr. Martin says. “So, when a farmer is dealing with this issue, it is a big problem for them.”

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A new wax coating technology for produce could provide enhanced protection against foodborne pathogens, according to new research publishd in Current Research in Food Science.

The idea was the brainstorm of Mustafa Akbulut, PhD, a professor in the department of chemical engineering at Texas A&M University in College Station, who teamed Luis Cisneros-Zevallos, PhD, a professor in the department of horticultural sciences at the same institution, to design longer-lasting and bacteria-free produce.

“Our collaborative research group combining cutting-edge engineering, food science, and horticulture science is actively engaged in developing new ideas to address issues related to food safety and shelf life,” Dr. Akbulut tells Food Quality & Safety. “In this work, we want to improve the wax that is already applied to many fruits and vegetables in order to enhance their cosmetics and reduce evaporative losses.”

Since a wax coating is already applied to produce, the team looked for a way to make food waxes more functional and beneficial to maximize their potential. “We wanted to create food waxes that have active and passive protection mechanisms against foodborne pathogens and spoilage microorganisms,” Dr. Akbulut says.

The need for this technology has several elements. First, food spoilage is an enormous burden to national economies worldwide. Even increasing the shelf-life of produce by one day can account for huge sums. This is important for sustainability and minimizing waste.

“Additionally, most food industry processes rely on sanitizers in the facility,” Dr. Akbulut says. “Usually, there is no protection after the food commodity leaves the facility. For instance, during transportation and display on grocery shelves, there are ample opportunities for bacterial contamination. At this point, let us imagine multiple people touching and selecting a food commodity from display shelves. Having a strategy to actively protect the food commodity even after it is sanitized and removed from the facility is extremely important.”

The new wax coating technology uses nano-encapsulated essential oils that are evenly distributed in food-grade wax. “The key is to design encapsulation systems that are compatible with the wax materials and can gradually release their contents,” Dr. Akbulut adds. “This method can extend the shelf life of fruits and vegetables by providing a sustained release of bioactive compounds.”

To date, the team have tested the coating against E. coli O157:H7 and S. aureus, demonstrating the product’s effectiveness against common contamination risks.

“It is a conformal coating,” he says. “It can be used for any produce as long as the produce is not very fragile or delicate. It can be commonly applied to many fruits and vegetables, including apples, stone fruits, citrus fruits, cucumbers, bell peppers, eggplants, and tomatoes. These are the starting products. Obviously, commodities more frequent association with foodborne outbreaks can benefit more from this technology.”

He believes that utilizing smarter protective wax coatings can directly translate to performance advantages in the marketplace and is a game changer for the produce industry because it can provide continuous protection of the food commodity even after it leaves the packing/grading facility; growers are already applying wax, so adding functional additives allows them to get additional benefits without major changes to operations; and, the produce will last longer.

The next steps for this technology are to apply it at a production scale to identify and resolve any scaling up issues in translating from lab bench prototypes to commercial packing line implementation. “Our eventual hope is to see this technology broadly implemented across a wide variety of fresh produce,” Dr. Akbulut says. “This would make the produce supply safer while extending shelf life to support sustainability efforts on a global scale by reducing food waste.”

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