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Troubleshooting problems with a small flock

When investigating a problem with a poultry flock, the following questions should be asked:

1. What are the type, quantity, age, and source of birds on the farm? It is impor-tant to know whether any poultry on the farm is of mixed species. Some diseases can be carried by one species showing no outward signs but can be lethal to other poultry species. For example, turkeys and peafowl are particularly susceptible to blackhead, which is cause by a protozoan parasite. While the free-living forms of the protozoa cannot survive long, they can be transmitted in the cecal worm, a parasite of chickens. For this reason, it is recommended that turkeys not be raised with chickens or on ground typically used for pasture-raised chickens.

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Fertilized vs. Non-fertile

The rooster must be present for an egg to be fertilized.

Fertilized vs. Non-fertile

Roosters are not necessary at egg farms where eggs are produced for human consumption and not incubation. Eggs for incubation are grown at special farms called breeder farms where roosters are present with the hens.

The next time you break open an egg; look for the germinal disc. You will see that supermarket eggs are infertile.

Did you know you can determine if an egg is fertile or not by looking at the germ spot? The germ spot is the white spot on the yolk. The non-fertile germ spot contains only the female’s cells and looks like a solid white spot. In a fertile egg the germ spot contains both the female and male cells. This allows cells to divide and the spot grows while the rest of the egg is being built in the female’s oviduct. Because of this growth the fertile germ spot on the yolk looks like a circle with a somewhat clear center.


Non-Fertile egg, Notice the Germinal spot just above center on the Yolk. It appears as solid white spot.


Fertile Egg. Notice the germinal spot looks larger and like a circle in this fertile egg.

ARM & HAMMER Adds New Bacillus Strains To Targeted Microbial Solutions

Source: ARM & HAMMER news release

Princeton, NJ – Arm & Hammer Animal and Food Production announces the discovery of two new beneficial Bacillus strains, Bacillus subtilis 839 and Bacillus subtilis 4976. The new strains will be incorporated into ARM & HAMMER Targeted Microbial Solutions available in CERTILLUS products for poultry, swine and cattle producers.

Both new strains have antimicrobial activity against harmful bacteria that are prevalent within the livestock and poultry industries. Bacillus subtilis 839 is effective against diverse E. coli species in both ruminants and poultry. Bacillus subtilis 4976 offers activity against E. coli, Salmonella, Clostridium perfringens and other clostridial bacteria that are common in poultry, swine, and beef and dairy cattle.

“Commercial development of these strains is integral to helping our customers address pathogenic disease and food safety threats,” says Dr. Xandra Smith, ARM & HAMMER manager of microbial ecology and genetics. “Through our commitment to research and problem-solving, livestock and poultry producers can support the overall production and health of their herds and flocks, while helping to protect the integrity of our food supply.”

Targeted Microbial Solutions are available exclusively from ARM & HAMMER through its CERTILLUS products for feed, forage and manure. These solutions are tailored to the individual operation based on an evaluation of the Microbial Terroir, an assessment of the microbial makeup of the environment, soil, animals and weather at the specific farm location. After identifying the operation’s unique challenges, ARM & HAMMER develops custom CERTILLUS solutions with the precise combination of Bacillus strains to address those challenges.

The recent addition of two new Bacillus strains to the Microbial Terroir portfolio is the result of continuous research to find new and more effective ways to expand coverage of the diverse pathogens that challenge livestock and poultry and threaten food safety. ARM & HAMMER researchers use advanced genetic testing tools to screen beneficial candidates from among its library of more than 25,000 bacterial isolates.

“As pathogens change and emerge, ARM & HAMMER is continually on the cusp of discovery to offer the most effective products. Developing new strains allows us to help producers fight food-borne pathogens as well as disease-causing pathogens that hamper animal efficiency and performance,” Smith says.

Poultry industry will be encouraged by study that finds the root of wooden breast syndrome

Source:University of DelawareSummary:The most delicious-looking broiler chicken could consist of hard, chewy meat. The problem plagues the poultry industry, but researchers have found the cause and mapped a solution. Gene expression irregularities at the onset of ‘wooden breast syndrome’ suggest the disease is a metabolic disorder. The findings could lead to short-term solutions that help growers manage the condition in chickens during production and may inform human health research related to metabolic syndromes, such as diabetes.

University of Delaware researchers have discovered that lipoprotein lipase, an enzyme crucial for fat metabolism, may be contributing to wooden breast syndrome in broiler chickens.

Wooden breast syndrome can affect broiler chickens, making the meat hard and chewy. It is a costly problem that can render the birds unmarketable, causing significant economic losses for growers, who sometimes see the disease in up to half their flocks.

The UD research team, led by Behnam Abasht, associate professor of animal and food sciences in UD’s College of Agriculture and Natural Resources, have identified gene expression irregularities at the onset of wooden breast syndrome that suggest the disease is a metabolic disorder characterized by abnormal fat accumulation in the breast muscle tissue.

The research team reported their results on Wednesday, Nov. 20 in the journal Scientific Reports.

The findings could help drive short-term solutions to help growers manage the condition in chickens at the production level through feed additives or supplements, or to reduce the number of birds that develop the condition.

“The industry desperately needs a solution right now. Conservative estimates project that wooden breast syndrome is costing the U.S. agricultural community $200 million dollars per year, but this number may be much higher,” said Abasht.

The United States leads production of broiler chickens worldwide. Delaware has more than 700 producers who raise commercial broiler chickens and over 1,000 small backyard poultry owners. Wooden breast syndrome is a concern around the world, too, as global consumption of chicken has risen in recent years as a leading source of dietary protein.

The research also could inform human health research related to metabolic syndromes, such as diabetes and atherosclerosis, which is associated with fatty deposits in the arteries.

Source of the problem

Abasht has been studying wooden breast syndrome in broiler chickens for nearly a decade. In previous work, his research team analyzed the genes involved in the disease and identified biomarkers for the disorder. They also characterized the unique biochemistry of the hardened breast tissue in chickens with the disease.

In their current work, the UD research team noticed that the expression of lipoprotein lipase was higher in chickens affected with wooden breast syndrome, leading more fat to accumulate in the chicken’s pectoral (or breast) muscles. Lipoprotein lipase serves as a “metabolic gatekeeper” that determines how much fat is allowed inside a given tissue.

This was curious because breast muscle fibers in chicken typically rely on sugar (glucose) molecules for fuel, not fat molecules.

The research team used RNA sequencing to determine which genes were being expressed in modern fast-growing broiler chickens and slower-growing legacy chickens. They then employed a new technology, called RNA in situ hybridization, to pinpoint exactly where this gene expression occurred inside the muscle.

The researchers found genetic evidence of lipoprotein lipase being expressed in endothelial cells in chickens, which was previously unknown. Present in all blood vessels, endothelial cells serve as a barrier between the blood and the surrounding tissue.

Abasht hypothesized that when more fat is oxidized for energy in the breast tissue of chickens, it may be causing the excessive release of free radical molecules that would modify (damage) fats and proteins in the muscle, prompting the chicken’s immune system to kick in to clear up the problem.

“We observed that if a particular vein was attacked by immune cells, the same vein typically also expressed higher levels of lipoprotein lipase,” said Abasht.

The researchers went a step further and compared this data with the gene expression signals found in two commercial broiler chicken lines, at three weeks of age and again at market age. In their analysis, the researchers found signals consistent with the fact that young chickens may be showing the same changes as market-age broiler chickens with wooden breast syndrome, even before the disease shows up.

The findings could provide potential markers to identify chickens that will develop the disease.

In recently published research in Genes, the researchers also reported finding common features between wooden breast syndrome in chickens and diabetes complications in humans, specifically diabetic cardiomyopathy, a chronic disease characterized by molecular and structural changes in the heart muscle.

Abasht’s research team is currently combing available literature for treating diabetes to see if there are ways to apply similar approaches to help broiler chickens avoid — or cope — with wooden breast syndrome, through diet, supplements or medication.

If they can find common solutions, Abasht said, a far-reaching goal of the work might be to use chickens as a model to study possible treatments for diabetes in humans.

“Our main focus was to address this problem from an agricultural perspective, but our findings open new horizons for future research that could benefit both agriculture and human health. This is a very interesting prospect for us,” said Abasht.

Longer term, Juniper Lake, a UD doctoral student in Abasht’s lab, said that having a deeper understanding of the genetic causes behind the disease could lead to solutions for agricultural producers to selectively breed out the traits that cause wooden breast disease, which has a relatively high heritability.

“There’s a lot to be gained from basic research of wooden breast, even if the main goal is to mitigate economic losses in the poultry industry,” said Lake.

With the Thanksgiving holiday just around the corner, it is important to note that wooden breast disease is currently only found in broiler chickens. It is not known to affect turkeys.

One Health Certified label seeks balanced approach to poultry production, marketing, By Don Ritter, DVM, director of technical marketing, Mountaire Farms

A new label under development for meat and poultry products will represent production that seeks a balanced approach to poultry production and marketing — one that ensures good health and welfare for the chickens while demonstrating responsible antibiotic usage under veterinary oversight to consumers, Don Ritter, DVM, director of technical marketing, Mountaire Farms, told Poultry Health Today.

Called One Health Certified, the new USDA Agricultural Marketing Service (USDA-AMS)-certified label is in part a response to arbitrary standards in production criteria, such as prohibiting the use of antibiotics to prevent, control or treat disease.

Ritter believes the supply chain from “no antibiotics ever” (NAE) production systems is not predictable nor sustainable. When the focus is only on never using antibiotics, the health of the animal suffers. There may be increased mortality. Those that survive require more feed and take longer to get to market, and the environment suffers too.

Okay but outdated

At the other end of the spectrum is what’s often referred to as “conventional” production, where veterinarians have the option of using all FDA-approved antibiotics for managing poultry diseases.  This approach to production has declined in recent years as poultry companies have sought to meet increasing consumer demand for chickens raised without antibiotics.

The One Health Certified label advocates responsible use of antibiotics, where needed, while giving consumers an audited, transparent program built upon five core pillars: biosecurity, veterinary care, antibiotic stewardship, animal welfare and environmental measurements.

The program has “action thresholds,” which make it unique, Ritter continued. For instance, when the health status of an animal group changes, actions are required. If it’s a disease process, there has to be veterinary involvement within 24 hours, and animals are properly taken care of. Everything is documented.

There are similar but not identical guidelines for all species. “I see it as an umbrella program…over the multiple protein classes,” he said.

Sustainable, practical

Nothing will fall through the cracks in this system, Ritter maintained, but he acknowledged it will require poultry companies to provide documentation about production practices.

The program is intended to avoid extra costs for consumers and has options that make it sustainable and practical, Ritter said. Mandating extreme practices for the entire supply chain that raise the cost of food takes away choice from people who don’t have resources.

Consumer research with about 1,000 people has indicated that 83% would buy products with the One Health Certified label. Respondents said they most valued veterinary care and want animals taken care of, he said.

Saturated NAE market

Asked if the trend toward NAE production can ever be reversed, considering it now accounts for over 50% of the US poultry market, Ritter said, “To me, gravity is a very powerful force, and I really do think the pendulum swung a little too far…” when that much of production became NAE.

Even though over 50% of chickens are produced now in NAE systems, NAE chicken meat accounts for only 10% of pounds sold, Ritter noted. NAE producers are selling 20% of their meat labeled as NAE and they’re getting a premium, but the meat is more expensive because producers have to recover the cost of running that type of production system. That premium is being eroded as this segment of the market becomes saturated, he said.

One Health Certified, which has been designed by a coalition that includes animal agriculture, nonprofit groups and university scientists, will require education and supportive messaging. The goal is to get it through the USDA-AMS auditing process and launch a website. The certification will be open to any producer who follows the rules, Ritter said.

Using neuroscience to improve the health and welfare of laying hens

A multi-million European-wide project to improve the conditions of egg-laying chickens has launched today, coinciding with World Animal Day.

We are just one of 20 partners working together as part of The ChickenStress European Training Network (ETN). This project will train future leaders in a range of scientific studies and help egg producers attain the best possible welfare standards for chickens. This project has received €3.9 million investment from the EU Marie Curie Fund.

Over the course of four years, 14 international studies, under the supervision of world experts in their fields, will be conducted to learn more about factors that contribute to stress response and resilience in poultry. The main goal of the project is to understand the stress response of chickens in various environments. A number of factors will be examined, including the impact of their housing system and feed program.

Dr. Tom Smulders from Newcastle University who is leading the project explained:

drtomsmulders.jpg

We will study how stress affects the brain and use this knowledge to identify best practice. We don’t know, for example, how the environment in which they are reared affects how well they adapt to the free-range systems they will be housed in in adulthood… These are the kind of welfare questions we want to answer with this project.

Dr. Tom Smulders
Newcastle University

A variety of methods will be used over the projects to measure stress responses and responsivity in laying hens, including markers in the brain (neurogenesis, different neurotransmitters), stress hormones, behavioral tests and observations, and health parameters.

Each project will involve a PhD student as part of the initiative to develop future scientific leaders. By working together with a range of experts, we are proud to be a part of this ambitious venture to maximize health and welfare of the birds in our care and throughout the industry.

All partners:

Newcastle University

Institut de Recerca Biomèdica de Lleida – Fundació Dr. Pifarré (IRBLleida)

Universidad de Lleida

The University of St-Andrews

The Open University of Israel

Universitaet Bern

Scotland’s Rural College

Universiteit Utrecht

Norwegian University of Life Sciences

Eigen Vermogen -Instituut voor Landbouw en Visserijonderzoek

Hendrix Genetics Research, Technology & Services BV

Trinity College Dublin

Vencomatic Group BV

Noldus Information Technology BV

Max Planck Institute for Ornithology

University of Veterinary Medicine, Vienna

Linköpings Universitet

The Lakes Free Range Co. Ltd.

Universiteit Gent

Tel Aviv University

Biosecurity: plans, puzzles and people

By Abby Neu Schuft, Extension Educator

NPIP Biosecurity Plan Audits

The count-down has begun for producers to complete their first NPIP Biosecurity Plan Audit. Each eligible producer needs to have their first audit complete by September 2020. There are many poultry industry members that still need to complete this process. There are also several who will complete their second audit this fall.

I would like to once again extend the invitation to you all that I am available to help you write your biosecurity plan and prepare for your first and subsequent NPIP audit. We can meet face-to-face, online or by phone. I understand putting pen to paper can be over-whelming, but I’m here to help. Please call (320) 235-0726 ext. 2019 or email (neux0012@umn.edu) and I’ll help you get started!

Biosecurity education and training opportunities

Many of you are familiar with or at least aware of the Biosecurity Entry Education Trailer (BEET) at the U. This educational tool has become so adaptable over the last four years! They are numerous ways to transform the trailer to accommodate any type of audience. In the past twelve months the trailer has been used in several unique ways:

Challengers had to solve four puzzles
within a time limit to escape the trailer,
all while learning about zoonotic disease
and biosecurity a the 2019 MN State Fair.

  • An escape room – an experiential learning opportunity during the 12 days of the 2019 MN State Fair
  • An interactive learning center for more than 350 MN 4-H youth participating in livestock programs
  • A prop, ‘test’ and discussion venue for 3rd year veterinary students attending the U of MN and Iowa State University
  • Set-up as a self-guided exhibit at events such as FarmFest, MN Pork Congress, and a world-wide family reunion hosted in Willmar, MN
  • Production system annual training for employees involved at every level of the company

Consider using BEET for annual employee training, as a physical model to design your new Danish Entry system, or to introduce youth to biosecurity concepts. Please contact us to plan your event!

Changes in cecum microbial community in response to total sulfur amino acid (TSAA: DL-methionine) in antibiotic-free and supplemented poultry birds

ABSTRACT

The effect of essential total sulfur amino acids (TSAA) like methionine and cysteine on the cecal microbiome of broilers was investigated at 2 different time points (days 21 and 42) of broiler rearing. A total of 360-day-old Cobb male broiler chicks were randomly distributed to 6 dietary treatments in a 2 × 3 factorial arrangement, with 2 levels of antibiotic growth promoters (AGP: 0 and 0.05%) and 3 levels of TSAA (DL-methionine) either for starter (0.7, 0.8, and 0.9%) or finisher chicks (0.52, 0.62, and 0.72%), labeled as diets 1 to 6. Cecal digesta from each replicate (n = 10) were sampled on days 21 and 42. DNA was extracted for the amplification of the V4 region of bacterial 16S rRNA genes and subjected to Illumina sequencing. Bioinformatic analyses were performed using QIIME, Mothur, and ad hoc tools and functional profiles of the inferred metagenome were analyzed using PICRUST. Statistical difference was determined by 2-way ANOVA and PERMANOVA. Clustering of cecal communities using PCoA showed clear separation of microbial communities based on age (P < 0.05) of birds and between low and medium/ high levels of TSAA (DL-methionine). At day 21, bacterial richness and diversity were higher than at day 42 where Clostridium cluster XI and Lactobacillus were found most abundant. No variability in taxonomic richness at the genus level was observed with AGP and DL-methionine supplementation. Interbird variation for richness was greater at day 42 compared to day 21. The mean fold difference of richness was greater (1.5 mean fold) with diets 1 and 6, suggesting interactive effects of AGP and TSAA (DL-methionine) in the diet. KEGG function profiles calculated by PICRUST suggest that the cecal microbiome increased glycolysis and energy generation correlated with increased dietary TSAA (DL-methionine) supplementation levels during the late broiler growth period (day 42). This study increases our knowledge of microbial dynamics and functions that are relevant to host nutrition and performance that may help us tailoring alternative strategies for raising poultry birds under antibiotic-free conditions.

BCO Lameness in Chickens: The Tip of an Iceberg

Group of broilers at a facility.

Bacterial chondronecrosis with osteomyelitis (BCO) is the most common cause of lameness in chickens in the United States, the European Union and South America. It affects clinically about 1.5% of all broilers grown to yield weights and can also affect turkeys and ducks.

Learn more: Strategies to Help Prevent or Manage BCO Lameness

Typically, 1.5% of broilers are culled from flocks that are affected by BCO lameness, but epidemic outbreaks can affect more than 15% of a flock. The highest incidences are in the fastest-growing flocks.

At the Zinpro International Poultry Seminar in Chicago, Dr. Adnan Alrubaye, DVM and Ph.D., Department of Poultry Science, and Biological Sciences, graduate program in Cell and Molecular Biology, University of Arkansas, gave a presentation about the impact of BCO lameness in chickens.

I caught up with him afterward, to discuss the details.

Marco Rebollo: How does BCO lameness work and how do broilers become infected?

Adnan Alrubaye: BCO lameness is caused by a variety of pathogenic microorganisms and stress factors, and somehow related to the growth of broilers, microfractures and developing circulation in the growth plates of the femur and tibia.

Bacteria can leak from the respiratory system and/or the gastrointestinal tract into the bloodstream and eventually settle in those growth plates, leading to necrosis. Eventually, the bones will weaken to the point that the bird can no longer stand or walk.

Even when clinical lameness is found in a low percentage, it indicates a larger number of other problems, such as damaged epithelial barriers (probably by coccidiosis, necrotic enteritis, dysbiosis, respiratory viruses, chronic respiratory disease, etc.), bacterial subclinical infections, potential food safety problems, and pain that compromises welfare.

MR: Are there ways to identify subclinical symptoms of BCO lameness in chickens?

AA: Currently, there is no identified method to diagnose the subclinical lesions in live birds, which is why BCO lameness is a serious issue. It is considered an important welfare issue that impacts the poultry industry in the U.S. and in many other countries across the world.

As a general rule, birds that experience pain due to BCO lameness try to hide their symptoms to avoid being attacked by other birds. As a result, it can be very difficult to identify any clinical symptoms of BCO lameness in chickens before it is too late. In many cases, we can only identify BCO lesions whenever birds start showing overt, clinical symptoms of lameness and/or during necropsy.

MR: You mentioned that there were other stressors that could lead to BCO lameness in chickens. Do you have any examples?

AA: There are many factors that can stress birds like rapid growth, handling, transportation and illnesses. We have been using the wire flooring model developed by emeritus professor, Dr. Robert Wideman, Department of Poultry Science, University of Arkansas, to create stress experimentally in order to induce lameness. In addition, we have developed a bacterial challenge with Staphylococcus agnetis (S. agnetis), that mimics the lameness epidemic for birds on litter flooring. The wire flooring model creates lameness in chickens by inducing a physical stress on long bones, which will ultimately lead to microfractures in the proximal femoral and tibial heads.  In addition, birds naturally prefer to walk on litter, so not allowing birds to have access to litter can increase their stress level and weakens the intestinal tight junctions, allowing bacteria to leak from the intestine into the bloodstream and, eventual colonization of the proximal femoral and tibial heads.

Learn More: Performance Trace Minerals Strengthen Tight Junctions

Using the wire flooring model or the bacterial challenge with S. agnetis can increase the incidence of lameness 50% to 70%.

MR: You conducted research studying the effect of supplementing broilers with Availa®ZMC on the incidence of lameness. What did you find?

AA: Feeding 1 kg of Availa-ZMC per metric ton of feed resulted in a significant reduction in BCO lameness in chickens and delayed the incidence by four to five days.

Two trials were conducted at the University of Arkansas, one using the wire flooring model and the other one on litter flooring incorporating BCO pathogen, S. agnetis isolate 908, to simulate a high virulence, natural lameness epidemic in a barn. Birds fed Availa-ZMC in the wire flooring trials experienced a reduction of up to 29% BCO lameness, while the birds fed Availa-ZMC in the S. agnetis challenge on litter trial showed a 23% reduction in the occurrence of BCO lameness. We also found that the birds fed Availa-ZMC had peripheral blood monocytes that were much more active in killing S. agnetis 908 in in vitro assays.

The data from these studies will be published in 2020.

The trace minerals in Availa-ZMC — zinc, manganese and copper — improve gut integrity by strengthening the tight junctions in the intestine so that bacteria won’t leak out and settle into microfractures in the growth plates. Availa-ZMC also helps improve bone quality and the overall health and immune function of the birds.

We’re happy to see this large of a reduction in the incidence of lameness in chickens when using Availa-ZMC, and we’re looking forward to doing more research with these products.

Performance Trace Minerals Reduce Lameness in Chickens

Improving gut health, strengthening tight junctions and improving bone strength are all key to preventing BCO lameness in chickens. To learn how performance trace minerals, like those found in Availa-ZMC, improve overall health in your poultry operation, contact your Zinpro representative today.

Salmonella’s sex life key to mitigating food safety risks, By Jean Guard, DVM, PhD, veterinary medical officer at the US Department of Agriculture’s Agricultural Research Service

Food safety challenges linked to Salmonella could be tackled more effectively by better understanding the sex life of bacteria, according to a leading US government veterinarian.

Researchers have known for some time that Salmonella and other bacteria have a sexual form of reproduction called homologous recombination.

However it seems the process, which sees bacteria swap sections of their chromosome with each other and ultimately create hybrid strains, could be much more common than initially believed.

Jean Guard, DVM, PhD, veterinary medical officer at the US Department of Agriculture’s Agricultural Research Service, said the development of diagnostic techniques and genome databases has led scientists to discover strains of Salmonella which don’t fit traditional bacterial categories.

By studying cellular differentiation — in other words, how bacteria cells change shape — she and her team have realized there are “worker” and “queen” bacteria, similar to bees in a hive.

And while the worker bacteria reproduce through binary fission, the queens have the potential to swap sections of genomes in a form of bacterial sex to produce new strains and new workers.

Speaking to Poultry Health Today at the American Association of Avian Pathologists conference, Guard said there are only a small number of queen bacteria in a bacterial population, which is why they have so far evaded detection.

Their low numbers also make it difficult to identify where hybrid Salmonella strains originate from, making it an important area for future research.

“We’re excellent at detecting workers which are out there infecting us, but what we’re not good at is recognizing the origin of all these new strains,” she said.

“So I feel like we’re caught in a cycle where we’re seeing Salmonella expand at certain times through the food safety system… but there has to be a cycle of chromosomal repair as all those workers get damaged and accumulate mutations.

“So what we’re not really figuring out is how does Salmonella repeat the repair of its chromosome to keep coming back around and around. And that’s a cycle I want to break.”

To take control of Salmonella — outbreaks of which haven’t shown any signs of reducing in humans over the past 15 years — Guard said more testing is needed across the food production system to identify the “oddball” bacteria that could be the source of hybrid strains.

“On farm and in the processing plant, what I would be looking for is areas that may be producing a variety of Salmonella serotypes,” she explained. “Where are you seeing unusual clusters of multiple serotypes emerging? Because that’s probably your nest.”

While the majority of these newcomer strains don’t survive, it just needs one hybrid to match with its environment before the bacteria can begin clonal expansion, she added.

On top of increased testing throughout the food chain, Guard also stressed the need to teach consumers about how to handle food properly.

“We can do everything that we do, and at the end of the day if the consumer doesn’t handle the product correctly, then they are adding to the problem. So education is always an important part of this picture.”

Poultry News

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