Evaluation of Liquid and Dry Chemical Treatments To Reduce Salmonella Typhimurium Contamination on Animal Food Manufacturing Surfaces.

ABSTRACT: Salmonella can be isolated from animal food, ingredients, and animal food manufacturing surfaces. There are limited data regarding the sanitation of animal food manufacturing surfaces. This experiment evaluated the effects of nine chemical treatments on reduction of Salmonella Typhimurium contamination on various manufacturing surfaces. This experiment was a 9 × 5 factorial with nine chemical treatments and five surfaces. The nine chemical treatments included one with no inoculation or sanitation treatment (negative control). In the other eight treatments, inoculation with Salmonella Typhimurium was followed by either no sanitation treatment (positive control) or treatment with ground corn; liquid commercial formaldehyde; liquid food-grade sanitizer; liquid medium chain fatty acid blend of caprylic, caproic, and capric acids (MCFA); dry commercial calcium propionate; dry commercial acidulant; and dry commercial benzoic acid. The five surfaces included stainless steel, plastic, polypropylene tote bag, rubber belt, and rubber tire. Plastic had higher levels of Salmonella in the positive control than did the polypropylene tote bag; other surfaces had intermediate levels (P < 0.05). Surfaces treated with formaldehyde had no detectable Salmonella after treatment, and surfaces treated with MCFA had at least a 4-log reduction compared to the control (P < 0.05). The dry acidulant was the most effective dry sanitizer tested, but it had no impact on Salmonella concentration on rubber tires (P < 0.05). Whereas liquid sanitizers were the most effective in this experiment, they have limitations for use in dry bulk systems. In summary, formaldehyde, food-grade sanitizer, and MCFA were the most effective chemical treatments to reduce Salmonella surface contamination. Surface type can also influence Salmonella mitigation strategies; specifically, stainless steel and plastic can be more challenging to sanitize within animal food facilities.

Effects of a dry acidulant addition to prevent Salmonella contamination in poultry feed.

Salmonella subs. serovar Enteritidis is a potential biological pathogen of concern in the poultry industry. Contamination of the bacterium on eggshells has led to human illnesses. With the implementation of new regulations, animal feed manufacturing continues to be under more stringent requirements. Specifically, there is zero tolerance for Salmonella Pullorum, Gallinarum, or Enteritidis in poultry feed. For this reason, it is important to determine an effective method of reducing or preventing Salmonella contamination in feed for poultry. Therefore, the objective of this study was to evaluate the impact of sodium bisulfate (SBS; Jones-Hamilton, Co., Walbridge, OH) added to poultry mash to reduce or prevent Salmonella growth over time. A single, commercially produced all-flock poultry mash was mixed with four different levels of SBS: 0.0%, 0.25%, 0.50%, and 0.70%. After SBS addition, the treated mash was inoculated with Salmonella enterica subsp, enterica serovar Enteritidis (ATCC 13076) and enumerated for Salmonella on days 0, 1, 2, 7, and 14 post-inoculation by plating on xylose lysine deoxycholate agar. There was no significant effect of SBS inclusion level on the reduction of Salmonella (P = 0.23); however, there was a significant effect of time across treatments (P < 0.0001). Additionally, there was no inclusion level × time interaction (P = 0.68). These results suggest that while SBS inclusion has no effect on Salmonella concentrations, storage time is effective at reducing or eliminating Salmonella contamination in poultry feed.

Research Note: Evaluating the roles of surface sanitation and feed sequencing on mitigating Salmonella Enteritidis contamination on animal food manufacturing equipment.

The objective of this study was to evaluate the efficacy of flushing surfaces with untreated feed vs. the use of 2 different dry chemical sanitizers on residual surface and feed Salmonella Enteritidis contamination. First, a Salmonella-negative batch of poultry feed was mixed in 9 laboratory-scale paddle mixers. A feed sample was collected, and targeted locations on surfaces within the mixer were swabbed to confirm Salmonella-negative. Next, a Salmonella-positive batch of poultry feed was mixed, sampled, and mixer surfaces swabbed. Mean Salmonella Enteritidis contamination across all 9 mixers were 3.63 cfu/g for sampled feed and 1.27 cfu/cm2 for surface contamination. Next, the mixers manufactured one of the following treatments (3 mixers/treatment): 1) none (control); 2) a commercially available essential oil blend; or 3) rice hulls treated with a 10% concentration of a propriety blend of medium-chain fatty acids (MCFA). After each treatment, each mixer manufactured another 2 batches of Salmonella-free feed (sequence 1 and sequence 2). Feed samples were collected, and surfaces were swabbed between each batch of feed. Manufacturing sequence (P < 0.0001) but not treatment (P > 0.05) impacted feed or surface contamination of Salmonella Enteritidis. There was Salmonella-positive residue in the batch of feed manufactured immediately after the positive control batch. However, no Salmonella residue was detected in batches of feed treated with either the commercial essential oil blend or MCFA. Low levels of Salmonella residue were observed from either feed (0.7 cfu/g for commercial essential oil blend) or surfaces (0.1 cfu/cm2 for MCFA) manufactured in sequence 1, but no residue was observed in sequence 2. These data suggest that sequencing of feed during manufacturing reduces Salmonella-positive contamination within animal food and on manufacturing surfaces, particularly after the second batch or with the use of chemical treatments.

Feed batch sequencing to decrease the risk of porcine epidemic diarrhea virus (PEDV) cross-contamination during feed manufacturing.

Feed has been identified as a vector of transmission for porcine epidemic diarrhea virus (PEDV). The objective of this study was to determine if feed batch sequencing methods could minimize PEDV cross-contamination. Porcine epidemic diarrhea virus-free swine feed was manufactured to represent the negative control. A 50 kg feed batch was mixed in a pilot scale feed mill for 5 min, sampled, then discharged for 10 min into a bucket elevator and sampled again upon exit. Next, a pathogenic PEDV isolate was used to inoculate 49.5 kg of PEDV-free feed to form the positive control. The positive control was mixed, conveyed and sampled similar to the negative control. Subsequently, 4 sequence batches (sequence 1 to 4) were formed by adding a 50 kg batch of PEDV-negative feed to the mixer after the prior batch was mixed and conveyed; all sequences were mixed, conveyed, and sampled similar to the negative and positive control batches. None of the equipment was cleaned between batches within a replicate. This entire process was replicated 3 times with cleaning the feed mill between replicates. Feed was then analyzed for PEDV RNA by real-time reverse transcriptase semiquantitative polymerase chain reaction (rRT-PCR) as measured by cycle threshold (Ct) and for infectivity by bioassay. Sequence 1 feed had higher (P ˂ 0.05) rRT-PCR Ct values than the positive batch and sequence 2 feed had higher (P ˂ 0.05) Ct values than sequence 1, regardless of sampled location. Feed sampled from the mixer from sequence 2, 3, and 4 was rRT-PCR negative whereas feed sampled from the bucket elevator was rRT-PCR negative from sequence 3 and 4. Bioassay was conducted using 66 mixed sex 10-d-old pigs confirmed negative for PEDV allocated to 22 different rooms. Pigs were initially 10-d old. Control pigs remained PEDV negative for the study. All pigs from the mixer positive batch (9/9) and bucket elevator positive batch (3/3) were rRT-PCR positive on fecal swabs by the end of the study. One replicate of pigs from mixer sequence 1 was rRT-PCR positive (3/3) by 7 dpi. One replicate of mixer pigs from sequence 2 was rRT-PCR positive (3/3) by 7 dpi although no detectable PEDV RNA was found in the feed. The results demonstrate sequenced batches had reduced quantities of PEDV RNA although sequenced feed without detectible PEDV RNA by rRT-PCR can be infectious. Therefore, a sequencing protocol can reduce but not eliminate the risk of producing infectious PEDV carryover from the first sequenced batch of feed.

Characterizing the rapid spread of porcine epidemic diarrhea virus (PEDV) through an animal food manufacturing facility.

New regulatory and consumer demands highlight the importance of animal feed as a part of our national food safety system. Porcine epidemic diarrhea virus (PEDV) is the first viral pathogen confirmed to be widely transmissible in animal food. Because the potential for viral contamination in animal food is not well characterized, the objectives of this study were to 1) observe the magnitude of virus contamination in an animal food manufacturing facility, and 2) investigate a proposed method, feed sequencing, to decrease virus decontamination on animal food-contact surfaces. A U.S. virulent PEDV isolate was used to inoculate 50 kg swine feed, which was mixed, conveyed, and discharged into bags using pilot-scale feed manufacturing equipment. Surfaces were swabbed and analyzed for the presence of PEDV RNA by quantitative real-time polymerase chain reaction (qPCR). Environmental swabs indicated complete contamination of animal food-contact surfaces (0/40 vs. 48/48, positive baseline samples/total baseline samples, positive subsequent samples/total subsequent samples, respectively; P < 0.05) and near complete contamination of non-animal food-contact surfaces (0/24 vs. 16/18, positive baseline samples/total baseline samples, positive subsequent samples/total subsequent samples, respectively; P < 0.05). Flushing animal food-contact surfaces with low-risk feed is commonly used to reduce cross-contamination in animal feed manufacturing. Thus, four subsequent 50 kg batches of virus-free swine feed were manufactured using the same system to test its impact on decontaminating animal food-contact surfaces. Even after 4 subsequent sequences, animal food-contact surfaces retained viral RNA (28/33 positive samples/total samples), with conveying system being more contaminated than the mixer. A bioassay to test infectivity of dust from animal food-contact surfaces failed to produce infectivity. This study demonstrates the potential widespread viral contamination of surfaces in an animal food manufacturing facility and the difficulty of removing contamination using conventional feed sequencing, which underscores the importance for preventing viruses from entering and contaminating such facilities.

Mitigation of Salmonella on Pet Food Kibbles by Using Liquid and Powdered 3-Hydroxy-3-Methylbutyric Acid.

In recent years, several pet food recalls have been attributed to Salmonella contamination. In addition to the negative impacts on animal health, Salmonella-contaminated pet foods have been linked to infection in humans. With that in mind, the U.S. Food and Drug Administration has set forth a zero-tolerance policy for Salmonella in pet foods. Typically, pet foods are extruded or processed at high temperatures that are sufficient to reduce pathogenic bacteria. However, the possibility for postextrusion contamination still exists. One potential method to reduce the risk of postextrusion contamination of pet foods with Salmonella is through the addition of a chemical additive coating. The objective of this research was to evaluate the ability of ß-hydroxy-ß-methylbutyrate (HMB), in either free acid (HMBFA) or calcium salt (CaHMB) form, to reduce postextrusion contamination of dry extruded dog kibble with Salmonella. Three trials were conducted with HMBFA and CaHMB coated onto the kibbles at levels of 0, 0.1, 0.3, 0.5, 0.9, and 1.5% (w/w). The coated kibbles were then inoculated with Salmonella enterica subsp. enterica Enteritidis (ATCC 13076), with enumeration done on days 0, 1, 2, 7, and 14 postinoculation. Subsamples on each day were serially diluted, spread plated to xylose lysine deoxycholate agar, and incubated at 37°C for 24 h. Salmonella colonies were then counted and log CFU per gram was calculated. The 1.5% HMBFA reduced counts by 4.9 ± 0.2 log units on day 1, whereas the positive control only decreased 2.2 ± 0.1 log units (P < 0.0001). The 1.5% CaHMB level decreased counts by 7.1 ± 0.04 log units by day 7 compared with the control decrease of 2.1 ± 0.1 log units (P < 0.0001). All HMBFA and CaHMB treatments resulted in the elimination of detectable Salmonella counts by day 14 (P < 0.0001 versus controls). In conclusion, HMB coating was effective at reducing Salmonella artificially inoculated to dog kibbles in a model of postextrusion contamination.

Elimination of Porcine Epidemic Diarrhea Virus in an Animal Feed Manufacturing Facility.

Porcine Epidemic Diarrhea Virus (PEDV) was the first virus of wide scale concern to be linked to possible transmission by livestock feed or ingredients. Measures to exclude pathogens, prevent cross-contamination, and actively reduce the pathogenic load of feed and ingredients are being developed. However, research thus far has focused on the role of chemicals or thermal treatment to reduce the RNA in the actual feedstuffs, and has not addressed potential residual contamination within the manufacturing facility that may lead to continuous contamination of finished feeds. The purpose of this experiment was to evaluate the use of a standardized protocol to sanitize an animal feed manufacturing facility contaminated with PEDV. Environmental swabs were collected throughout the facility during the manufacturing of a swine diet inoculated with PEDV. To monitor facility contamination of the virus, swabs were collected at: 1) baseline prior to inoculation, 2) after production of the inoculated feed, 3) after application of a quaternary ammonium-glutaraldehyde blend cleaner, 4) after application of a sodium hypochlorite sanitizing solution, and 5) after facility heat-up to 60°C for 48 hours. Decontamination step, surface, type, zone and their interactions were all found to impact the quantity of detectable PEDV RNA (P < 0.05). As expected, all samples collected from equipment surfaces contained PEDV RNA after production of the contaminated feed. Additionally, the majority of samples collected from non-direct feed contact surfaces were also positive for PEDV RNA after the production of the contaminated feed, emphasizing the potential role dust plays in cross-contamination of pathogen throughout a manufacturing facility. Application of the cleaner, sanitizer, and heat were effective at reducing PEDV genomic material (P < 0.05), but did not completely eliminate it.

Evaluating Chemical Mitigation of Salmonella Typhimurium ATCC 14028 in Animal Feed Ingredients.

Salmonella Typhimurium is a potential feed safety hazard in animal feed ingredients. Thermal mitigation of Salmonella spp. during rendering is effective but does not eliminate the potential for cross-contamination. Therefore, the objective of this experiment was to evaluate the effectiveness of chemicals to mitigate postrendering Salmonella Typhimurium ATCC 14028 contamination in rendered proteins over time. Treatments were arranged in a 6 × 4 factorial with six chemical treatments and four rendered protein meals. The chemical treatments included (i) control without chemical treatment, (ii) 0.3% commercial formaldehyde product, (iii) 2% essential oil blend, (iv) 2% medium chain fatty acid blend, (v) 3% organic acid blend, and (vi) 1% sodium bisulfate. The four rendered protein meals included (i) feather meal, (ii) blood meal, (iii) meat and bone meal, and (iv) poultry by-product meal. After matrices were chemically treated, they were inoculated with Salmonella Typhimurium ATCC 14028, stored at room temperature, and enumerated via plate counts on days 0, 1, 3, 7, 14, 21, and 42 postinoculation. The Salmonella concentration in ingredients treated with medium chain fatty acid and commercial formaldehyde were similar to one another (P = 0.23) but were 2 log lower than the control (P < 0.05). Ingredients treated with organic acids and essential oils also had lower Salmonella concentrations than the control (P < 0.05). Time also played a significant role in Salmonella mitigation, because all days except days 14 and 21 (P = 0.92) differed from one another. Rendered protein matrix also affected Salmonella stability, because concentrations in meat and bone meal and blood meal were similar to one another (P = 0.36) but were greater than levels in feather meal and poultry by-product meal (P < 0.05). In summary, chemical treatment and time both mitigated Salmonella Typhimurium ATCC 14028, but their effectiveness was matrix dependent. Time and chemical treatment with medium chain fatty acids or a commercial formaldehyde product were most effective at mitigating Salmonella Typhimurium ATCC 14028 in rendered protein meals.

Evaluation of a Biological Pathogen Decontamination Protocol for Animal Feed Mills.

Animal feed and ingredients are potential vectors of pathogenic bacteria. Contaminated ingredients can contaminate facility equipment, leading to cross-contamination of other products. This experiment was conducted to evaluate a standardized protocol for decontamination of an animal feed manufacturing facility using Enterococcus faecium (ATCC 31282) as an indicator. A pelleted swine diet inoculated with E. faecium was manufactured, and environmental samples (swabs, replicate organism detection and counting plates, and air samples) were collected (i) before inoculation (baseline data), (ii) after production of inoculated feed, (iii) after physical removal of organic material using pressurized air, (iv) after application of a chemical sanitizer containing a quaternary ammonium-glutaraldehyde blend, (v) after application of a chemical sanitizer containing sodium hypochlorite, (vi) after facility heat-up to 60 8 C for 24 h, (vii) for 48 h, and (viii) for 72 h. Air samples collected outside the facility confirmed pathogen containment; E. faecium levels were equal to or lower than baseline levels at each sample location. The decontamination step and its associated interactions were the only variables that affected E. faecium incidence (P < 0.0001 versus P > 0.22). After production of the inoculated diet, 85.7% of environmental samples were positive for E. faecium. Physical cleaning of equipment had no effect on contamination (P = 0.32). Chemical cleaning with a quaternary ammonium-glutaraldehyde blend and sodium hypochlorite each significantly reduced E. faecium contamination (P < 0.0001) to 28.6 and 2.4% of tested surfaces, respectively. All samples were negative for E. faecium after 48 h of heating. Both wet chemical cleaning and facility heating but not physical cleaning resulted in substantial E. faecium decontamination. These results confirmed both successful containment and decontamination of biological pathogens in the tested pilot-scale feed mill.