Category Archives: Research

Research – Intensive farming increases risk of epidemics

Science Daily

Overuse of antibiotics, high animal numbers and low genetic diversity caused by intensive farming techniques increase the likelihood of pathogens becoming a major public health risk, according to new research led by UK scientists.

An international team of researchers led by the Universities of Bath and Sheffield, investigated the evolution of Campylobacter jejuni, a bacterium carried by cattle which is the leading cause of gastroenteritis in high income countries.

Campylobacter facts:

  • Causes bloody diarrhea in humans
  • Transferred to humans from eating contaminated meat and poultry
  • Although not as dangerous as typhoid, cholera or E.coli, it causes serious illness in patients with underlying health issues and can cause lasting damage.
  • Around 1 in 7 people suffer from an infection at some point in their life
  • Causes three times more cases than E.coli, Salmonella and listeria combined
  • Carried in the faeces of chickens, pigs, cattle and wild animals
  • Campylobacter is estimated to be present in the faeces of 20% cattle worldwide
  • The bug is very resistant to antibiotics due to their use in farming

The researchers, publishing in the journal Proceedings of the National Academy of Sciences, studied the genetic evolution of the pathogen and found that cattle-specific strains of the bacterium emerged at the same time as a dramatic rise in cattle numbers in the 20th Century.

The authors of the study suggest that changes in cattle diet, anatomy and physiology triggered gene transfer between general and cattle-specific strains with significant gene gain and loss. This helped the bacterium to cross the species barrier and infect humans, triggering a major public health problem.

Combine this with the increased movement of animals globally, intensive farming practices have provided the perfect environment in which to spread globally through trade networks.

Professor Sam Sheppard from the Milner Centre for Evolution at the University of Bath, said: “There are an estimated 1.5 billion cattle on Earth, each producing around 30 kg of manure each day; if roughly 20 per cent of these are carrying Campylobacter, that amounts to a huge potential public health risk.

“Over the past few decades, there have been several viruses and pathogenic bacteria that have switched species from wild animals to humans: HIV started in monkeys; H5N1 came from birds; now Covid-19 is suspected to have come from bats.

“Our work shows that environmental change and increased contact with farm animals has caused bacterial infections to cross over to humans too.

“I think this is a wake-up call to be more responsible about farming methods, so we can reduce the risk of outbreaks of problematic pathogens in the future.”

Professor Dave Kelly from the Department of Molecular Biology and Biotechnology at the University of Sheffield said: “Human pathogens carried in animals are an increasing threat and our findings highlight how their adaptability can allow them to switch hosts and exploit intensive farming practices.”

The researchers hope that their study can help scientists predict potential problems in the future so they can be prevented before they turn into another epidemic.

Research – Fusarium: How to assess and manage the risk in cereals

FWI

Protecting crops against the damaging effects of fusarium ear blight is essential in any year, but it could be even more so this season, given the reduced wheat area and strong premiums for milling wheat.

Fusarium and its associated mycotoxins are not just a concern for milling wheat growers, though, as the different forms of the disease can affect yield and quality in all the main cereals: wheat, barley and oats.

As there are legal mycotoxin limits on grain for human consumption, and guidance limits on crops intended for animal feed, it is vital to protect all crops.

Research – Effectiveness and Functional Mechanism of a Multicomponent Sanitizer against Biofilms Formed by Escherichia coli O157:H7 and Five Salmonella Serotypes Prevalent in the Meat Industry

Journal of Food Protection

ABSTRACT

Biofilm formation by Escherichia coli O157:H7 and Salmonella enterica at meat processing plants poses a potential risk of meat product contamination. Many common sanitizers are unable to completely eradicate biofilms formed by these foodborne pathogens because of the three-dimensional biofilm structure and the presence of bacterial extracellular polymeric substances (EPSs). A novel multifaceted approach combining multiple chemical reagents with various functional mechanisms was used to enhance the effectiveness of biofilm control. We tested a multicomponent sanitizer consisting of a quaternary ammonium compound (QAC), hydrogen peroxide, and the accelerator diacetin for its effectiveness in inactivating and removing Escherichia coli O157:H7 and Salmonella enterica biofilms under meat processing conditions. E. coli O157:H7 and Salmonella biofilms on common contact surfaces were treated with 10, 20, or 100% concentrations of the multicomponent sanitizer solution for 10 min, 1 h, or 6 h, and log reductions in biofilm mass were measured. Scanning electron microscopy (SEM) was used to directly observe the effect of sanitizer treatment on biofilm removal and bacterial morphology. After treatment with the multicomponent sanitizer, viable E. coli O157:H7 and Salmonella biofilm cells were below the limit of detection, and the prevalence of both pathogens was low. After treatment with a QAC-based control sanitizer, surviving bacterial cells were countable, and pathogen prevalence was higher. SEM analysis of water-treated control samples revealed the three-dimensional biofilm structure with a strong EPS matrix connecting bacteria and the contact surface. Treatment with 20% multicomponent sanitizer for 10 min significantly reduced biofilm mass and weakened the EPS connection. The majority of the bacterial cells had altered morphology and compromised membrane integrity. Treatment with 100% multicomponent sanitizer for 10 min dissolved the EPS matrix, and no intact biofilm structure was observed; instead, scattered clusters of bacterial aggregates were detected, indicating the loss of cell viability and biofilm removal. These results indicate that the multicomponent sanitizer is effective, even after short exposure with dilute concentrations, against E. coli O157:H7 and S. enterica biofilms.

HIGHLIGHTS
  • No viable biofilm cells were detected after treatment with the multicomponent sanitizer.
  • Prevalence of both pathogens was low after treatment with the multicomponent sanitizer.
  • SEM analysis revealed that treatment dissolved the EPS matrix and destroyed the biofilm.

Research – Examination of the Culturable Microbiota from Low-Moisture Foods Imported into Canada for Antibacterial Activity against Listeria monocytogenes

Journal of Food Protection

ABSTRACT

Listeria monocytogenes, a resilient and ubiquitous foodborne pathogen, is associated with a high case-fatality rate in humans. This study investigated the culturable microbiota of low-moisture foods (LMFs) imported into Canada to see how well bacteria isolated from these foods could inhibit or inactivate the growth of L. monocytogenes. Imported LMFs were acquired from various supermarkets in the Greater Toronto Area (Ontario, Canada). The foods included dried apples, bee pollen, cumin seeds, date fruits, fennel seeds, pistachios, raisins, and seaweed. Bacterial strains were isolated from the foods using blood agar and then screened using an in-house-designed growth inhibition plate assay against L. monocytogenes. The inhibitory strains detected were then identified using 16S rRNA sequencing. Diverse bacteria were recovered from the foods; 236 isolates belonging to 122 observed phenotypes were obtained. From the inhibition plate assays, 10 of the 11 imported LMFs harbored inhibitory strains against L. monocytogenes, whereby 48 of the collected isolates (20%) were found to produce a zone of inhibition against this pathogen. The inhibitory strains belonged to six genera (Acinetobacter, Aerococcus, Bacillus, Lysinibacillus, Paenibacillus, and Sporosarcina) and 15 unique species. Among all foods tested, the date fruit microbiota displayed the greatest number and diversity of anti–L. monocytogenes inhibitory strains. Overall, it was found that the culturable microbiota of LMFs, imported into Canada, possess bacterial members that can inhibit the growth of L. monocytogenes. These results could lead to the discovery of either novel antimicrobial metabolites or beneficial anti–L. monocytogenes bacteria that could be added to foods to inactivate and/or control L. monocytogenes.

HIGHLIGHTS
  • Modification to an inhibition plate assay detects anti-Listeria bacteria more rapidly.
  • Unconventional anti-Listeria genera, such as Lysinibacillus, were discovered.
  • Date fruit microbiota warrants further investigation for anti-Listeria applications.
  • Isolated bacteria from low-moisture foods are diverse and can inhibit L. monocytogenes growth.

Research – Inhibitory effect of thyme and cinnamon essential oils against E. coli O157:H7 in Tahini

Scielo

Tahini is a common food product in the Mediterranean area that is used as a main ingredient in variety of ready-to-eat foods. The objective of the current study was to investigate the inhibitory effect of thyme oil (TO) or cinnamon oil (CO) on E. coli O157:H7 viability in tahini and diluted tahini at different storage temperatures. Addition of 2.0% CO to tahini reduced E. coli O157:H7 numbers by 1.38, 1.79 or 2.20 log10 CFU/mL at 10, 25 or 37 °C, respectively, by 28d. In diluted tahini at 10 °C, no viable cells of E. coli O157:H7 by 21d were detected when 1.0% CO was used. However, at 25 and 37 °C, no viable cells were detected by 14d when CO was added at 0.5% level. Addition of 2.0% TO to tahini, resulted in 1.82, 2.01 or 1.65 log10 CFU/mL reduction in E. coli O 157:H7 numbers was noted at 37, 25 or 10 °C, respectively, by 28d. In diluted tahini, TO at 0.5% or 1.0% induced complete reduction in the viability of E. coli O157:H7 by 28d storage at 37 or 25 °C. At 10 °C, a 3.02 log10 CFU/mL reduction was observed by 28d compared to the initial inoculation level in samples treated with 2.0% TO.

Research – Presence of Bacterial Pathogens and Levels of Indicator Bacteria Associated with Duck Carcasses in a Commercial Processing Facility

Journal of Food Protection

ABSTRACT

Little information has been published on the microbiological aspects of U.S. commercial duck processing. The objective of this study was to measure prevalence and/or levels of bacteria in duck samples representing the live bird and partially or fully processed oven-ready duck meat. At 12 monthly sampling times, samples were collected at six sites along the processing line in a commercial duck slaughter plant. Crop and cecum samples were collected at the point of evisceration. Whole carcass rinse samples were collected before and after carcass immersion chilling plus application of an antimicrobial spray. Leg quarters were collected from the cut-up line before and after application of an antimicrobial dip treatment. All samples (five from each site per monthly replication) were directly plated and/or enriched for Salmonella and Campylobacter. For the last 10 replications, carcass and leg quarter rinse samples were also evaluated for enumeration of total aerobic bacteria, Escherichia coli, and coliforms. Most cecum, crop, and prechill carcass rinse samples were positive for Campylobacter (80, 72, and 67%, respectively). Carcass chilling and chlorinated spray significantly lowered Campylobacter prevalence (P < 0.01), and even fewer leg quarters were positive for Campylobacter (P < 0.01). Passage through a chlorinated dip did not further reduce Campylobacter prevalence on leg quarters. Salmonella was infrequently found in any of the samples examined (≤10%). Total aerobic bacteria, coliforms, and E. coli levels were reduced (P < 0.01) on whole carcasses by chilling but were not different after cut-up or leg quarter dip treatment. Overall, current commercial duck processing techniques as applied in the tested plant were effective for reducing the prevalence and levels of Campylobacter on duck meat products.

HIGHLIGHTS
  • Campylobacter and Salmonella can be associated with live ducks for commercial slaughter.
  • Antimicrobial treatment reduced the prevalence of Campylobacter in duck samples.
  • Antimicrobial treatment reduced levels of aerobic bacteria, coliforms, and E. coli in duck samples.

Research – High-Pressure Inactivation of Histamine-Forming Bacteria Morganella morganii and Photobacterium phosphoreum

Journal of Food Protection

ABSTRACT

The effects of high hydrostatic pressure (HHP) treatments on histamine-forming bacteria (HFB) Morganella morganii and Photobacterium phosphoreum in phosphate buffer and tuna meat slurry were investigated using viability counting and scanning electron microscopy. The first-order model fits the destruction kinetics of high pressure on M. morganii and P. phosphoreum during the pressure hold period. The D-values of M. morganii (200 to 600 MPa) and P. phosphoreum (100 to 400 MPa) in phosphate buffer ranged from 16.4 to 0.08 min and 26.4 to 0.19 min, respectively, whereas those in tuna meat slurry ranged from 51.0 to 0.09 min and 71.6 to 0.19 min, respectively. M. morganii had higher D-values than P. phosphoreum at the same pressure, indicating it was more resistant to HHP treatment. HFB had a higher D-value in tuna meat slurry compared with that in phosphate buffer, indicating that the HFB were more resistant to pressure in tuna meat slurry. The Zp values (pressure range that results in a 10-fold change in D-value) of M. morganii and P. phosphoreum were 162 and 140 MPa in phosphate buffer and 153 and 105 MPa in tuna meat slurry, respectively. Damage to the cell wall and cell membrane by HHP treatments can be observed by scanning electron microscopy. To our knowledge, this is the first report to demonstrate that HHP can be applied to inactivate the HFB M. morganii and P. phosphoreum by inducing morphological changes in the cells.

HIGHLIGHTS
  • M. morganii with higher D-values was more resistant than P. phosphoreum at the same HHP.
  • HFB with higher D-values were more resistant in fish slurry than in phosphate buffer.
  • P. phosphoreum with lower Zp values was more sensitive to changes in HHP than M. morganii.
  • SEM shows that HHP causes cell wall and membrane damage in HFB.

Research – Effect of Peracetic Acid Solutions and Lactic Acid on Microorganisms in On-Line Reprocessing Systems for Chicken Slaughter Plants

Journal of Food Protection

ABSTRACT

During poultry slaughter and processing, microbial cross-contamination between individual chickens is possible, as well as from one slaughter animal to the next without direct contact. One option for reducing the risk of cross-contamination is to decrease the number of microorganisms on contact surfaces by using disinfectants. The aim is to decontaminate the surfaces coming into direct contact with the carcasses. In the present study, the effectiveness of different disinfectants was investigated in laboratory settings, simulating the conditions in the slaughterhouses and in a chicken slaughterhouse. For this, an artificial residue substance (consisting of yeast extract, albumin, and agar) was developed, tested, and included in the assays. Two disinfectants were tested under laboratory conditions: lactic acid (5 and 6.67%) and peracetic acid (0.33 and 0.5%). At the slaughterhouse, peracetic acid (0.021%) was used. In the laboratory tests, it was found that the peracetic acid solution had the highest disinfection potential with respect to an Escherichia coli strain (reduction >4 log CFU mL−1) at 0.5% without an artificial residue substance. The tested lactic acid solutions also showed the highest disinfection potential against a Pseudomonas aeruginosa strain, without an artificial residue substance. When applying the artificial residue substance, the reduction potential of lactic acid and peracetic acid was decreased to less than 1.4 log CFU mL−1. Application of peracetic acid in the slaughterhouse reduced the number of total aerobic bacteria by more than 4 log CFU mL−1 and the number of Enterobacteriaceae by more than 3 log CFU mL−1, depending on the place of sampling.

HIGHLIGHTS
  • Peracetic acid and lactic acid decreases E. coli and P. aeruginosa numbers in vitro.
  • Sanitation in place reduces the number of bacteria in a chicken slaughterhouse.
  • The number of total aerobic bacteria and Enterobacteriaceae was significantly reduced.

Research – Two Generally Recognized as Safe Surfactants plus Acidulants Inactivate Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes in Suspension or on Dip-Inoculated Grape Tomatoes

Journal of Food Protection

ABSTRACT

Contamination of fresh produce with the foodborne pathogens Salmonella enterica, Listeria monocytogenes, and Escherichia coli O157:H7 continues to be problematic, resulting in outbreaks of foodborne illness and costly corporate recalls. Various individual concentrations of citric or lactic acids (0.35 to 0.61%) or isopropyl citrate (0.16 to 0.54%) combined with two generally recognized as safe surfactants, 0.025% sodium-2-ethyl-hexyl sulfate and 0.025% sodium dodecylbenzene-sulfonate, were tested against these three pathogens in suspension and when inoculated and dried on the surface of grape tomatoes. The efficacy of sodium hypochlorite (NaClO; at 46 ppm) was also evaluated under dirty and clean conditions in suspension after addition of 0.3 or 0.03% bovine serum albumin, respectively, as an organic load. NaClO (46 ppm) inactivated the three pathogens in suspension by <0.76 log CFU/mL after 5 min in the presence of 0.3% bovine serum albumin, whereas 9 and 15 ppm of free chlorine inactivated the pathogens by 0.64 and 2.77 log CFU/mL, respectively, after 5 min under clean conditions. Isopropyl citrate (0.16% acidulant) plus 0.05% total concentration of the two surfactants inactivated the pathogens in suspension by up to 7.0 log CFU/mL within 2 min. When applied to grape tomatoes for 2 min, 0.54% isopropyl citrate plus 0.025% concentrations of each of the two surfactants reduced Salmonella, E. coli O157:H7, and L. monocytogenes by as much as ca. 5.47, 4.89, and 4.19 log CFU/g, respectively. These reductions were significantly greater than those achieved with 49 ppm of free chlorine. Citric acid and lactic acid plus surfactant washes achieved greater inactivation than water-only washes, reducing Salmonella, E. coli O157:H7, and L. monocytogenes on tomatoes by up to 4.90, 4.37, and 3.98 log CFU/g, respectively. These results suggest that these combinations of acidulants and surfactants may be an effective tool for preventing cross-contamination during the washing of grape tomatoes, for reducing pathogens on the fruit itself, and as an alternative to chlorine for washing fresh produce.

HIGHLIGHTS
  • Acidulants and surfactants were used in combinations against three foodborne pathogens.
  • Acidulant (0.16%) plus surfactants (0.05%) inactivated pathogens in suspension by up to 7.0 log CFU/mL.
  • Acidulant plus surfactant inactivated pathogens on dip-inoculated grape tomatoes by 3.35 to 5.47 log CFU/g.

Research – Phage Biocontrol Improves Food Safety by Significantly Reducing the Level and Prevalence of Escherichia coli O157:H7 in Various Foods

Journal of Food Protection

ABSTRACT

Management of Shiga toxin–producing Escherichia coli (STEC), including E. coli O157:H7, in food products is a major challenge for the food industry. Several interventions, such as irradiation, chemical disinfection, and pasteurization, have had variable success controlling STEC contamination. However, these interventions also indiscriminately kill beneficial bacteria in foods, may impact organoleptic properties of foods, and are not always environmentally friendly. Biocontrol using bacteriophage-based products to reduce or eliminate specific foodborne pathogens in food products has been gaining attention due to the specificity, safety, and environmentally friendly properties of lytic bacteriophages. We developed EcoShield PX, a cocktail of lytic bacteriophages, that specifically targets STEC. This study was conducted to examine the efficacy of this bacteriophage cocktail for reducing the levels of E. coli O157:H7 in eight food products: beef chuck roast, ground beef, chicken breast, cooked chicken, salmon, cheese, cantaloupe, and romaine lettuce. The food products were challenged with E. coli O157:H7 at ca. 3.0 log CFU/g and treated with the bacteriophage preparation at ca. 1 × 106, 5 × 106, or 1 × 107 PFU/g. Application of 5 × 106 and 1 × 107 PFU/g resulted in significant reductions (P < 0.05) in E. coli O157:H7 levels of up to 97% in all foods. When bacteriophages (ca. 1 × 106 PFU/g) were used to treat lower levels of E. coli O157:H7 (ca. 1 to 10 CFU/10 g) on beef chuck roast samples, mimicking the levels of STEC found under real-life conditions in food processing plants, the prevalence of STEC in the samples was significantly reduced (P < 0.05) by ≥80%. Our results suggest that this STEC-targeting bacteriophage preparation can result in significant reduction of both the levels and prevalence of STEC in various foods and, therefore, may help improve the safety and reduce the risk of recalls of foods at high risk for STEC contamination.

HIGHLIGHTS
  • Shiga toxin–producing E. coli is a major food safety challenge for the food industry.
  • EcoShield PX bacteriophage cocktail reduced E. coli O157:H7 levels in foods by up to 97%.
  • The bacteriophage cocktail reduced the prevalence of E. coli O157:H7 in foods by ≥80%.