Dry Pet Food Research Articles

Predictive equations of selenium accessibility of dry pet foods.

The trace element selenium is essential to both dogs and cats. Dry diets are formulated with a large range of ingredients, which may vary in selenium concentration and accessibility. This paper reports equations to predict the average invitro selenium accessibility from dry pet foods based on essential dietary nutrient concentrations, including crude protein, amino acids and crude fat. Predictive equations were made using stepwise linear regression for extruded and pelleted diets. The equations can be used to aid diet formulation to optimize selenium accessibility within the diet and to prevent selenium deficiency or toxicity.

Assessing the potential for Salmonella growth in rehydrated dry dog food

A substantial percentage of dog owners add water to dry dog food to increase its palatability. The recent association of Salmonella contamination of dry pet foods with salmonellosis cases in both dogs and their owners has generated a need to determine the ability of Salmonella to grow in eight commercial brands of rehydrated dry dog food. Eight brands of commercial dry dog food were rehydrated to 20, 35 and 50% added moisture, inoculated with two S. enterica strains (~105 CFU/g) and incubated for 72 h at 18 °C, 22 °C, or 28 °C. Dog food brand, moisture content, and temperature affected pathogen growth/survival patterns. Rehydration to 20% moisture did not support growth of S. enterica, and in general there was a 0.5–2.0 Log decline. At 35% moisture and 28 °C, 4 of 8 brands supported up to 3.4 Log(CFU/g) of growth, while Salmonella levels declined in three brands, and remained unchanged in one. Rehydration to 50% moisture at 28 °C supported increases of up to 4.6 Log(CFU/g) in 5 of 8 brands. Growth kinetics determinations with two of the brands that supported growth had calculated lag times, generation times, and maximum population densities of 4.4 and 2.2 h, 1.4 and 10.8 h, and 7.3 and 6.9 Log(CFU/g) when rehydrated to 35% moisture and held at 30 °C. Results of this study establish that the rehydration of dry dog food with sufficient amounts of water may support the growth of S. enterica. Based on the most rapid observed lag times, growth of Salmonella, if present, in rehydrated dog food could be avoided by discarding or refrigerating uneaten portions within 2–3 h of rehydration. These data allow accidental or intentional rehydration of dry dog food to be factored into predictive microbiology models and exposure assessments.

Development and Validation of an ED-XRF Method for the Fast Quantification of Mineral Elements in Dry Pet Food Samples

A simple and fast method for the quantification of a series of macro elements (Na, Mg, P, Cl, K and Ca) and trace elements (Mn, Fe, Cu and Zn) by Energy Dispersive X-Ray Fluorescence (ED-XRF) was developed and validated using samples of commercial dry pet foods. All samples were systematically analyzed by reference methods (i.e. Potentiometry for chloride and Inductively coupled plasma-atomic emission spectroscopy (ICP-AES) for other elements) and by ED-XRF. A calibration set of 33 samples was selected to uniformly cover the concentration ranges of each element and their reference method values were used to calibrate the ED-XRF device. A second set (the validation set) was composed of 46 other samples, which were analyzed as unknowns. During this validation step, the elemental concentrations obtained by ED-XRF were systematically evaluated against those obtained from the reference methods. Using robust statistics, for the 10 investigated analytes, the bias between both methods was found to be not significantly different from 0 and the relative uncertainties RSD(u) were found to be lower than 15 %. This study demonstrated that ED-XRF is a fast and reliable method for minerals quantification in dry pet foods.

Development and inter-laboratory study of a method for quantifying zearalenone in pet foods

An analytical method for quantifying zearalenone (ZEA) in pet foods using a liquid chromatograph-atmospheric pressure chemical ionization-mass spectrometer (LC-APCI-MS) was developed, and an inter-laboratory study was conducted. ZEA was extracted from pet food samples with aqueous acetonitrile. The extract was purified using a multifunctional column, MultiSep 226 Aflazon+, and analysed with LC-APCI-MS. The limit of quantification of ZEA by this method was estimated to be 0.2 mg/kg for dry and semi-dry pet foods, and 0.1 mg/kg for wet pet food. The calibration curve of ZEA for this method showed linearity in the range of 0.1-10 ng of ZEA (0.01-1.0 µg/ml, 0.04-4.0 mg/kg matrix). The mean recoveries, repeatabilities, reproducibilities, and the HorRats for ZEA from the inter-laboratory validation study were 99.0-102%, 2.2-3.0%, 5.6-6.6%, and 0.33-0.36, respectively.

ВПЛИВ УМОВ ЗБЕРІГАННЯ НА ПОЖИВНІСТЬ СУХИХ КОРМІВ ДЛЯ НЕПРОДУКТИВНИХ ТВАРИН

Stability in storage of dry feed and of their storage duration without appreciably reducing the nutritional value depends not only on the environmental factors, but also on the formulation and production technology. Humidity, temperature contribute to the development of microorganisms that consume energy and nutrients for the development, resulting in reduced energy content therein and crude protein. Manufacturers of dry pet food to ensure feed quality and slowing deterioration and development of microbial flora of added preservatives and antioxidants of different origin, which break the chain of oxidation. The introduction of inhibitors in an amount of 0.01% fat oxidation resistance increases 10...15 times.The performed investigations have established that during storage of dry pet food in a variety of storage conditions reduced the content of crude protein, crude fat and increased humidity, despite the conservation of feed. The greatest damage to the nutrients mentioned for the «summer conditions» storage in dry pet food which preserved natural ingredients, which is explained by the most intensive development of microflora in these conditions, and the oxidation of fat under the influence of high temperature and humidity. The best preserved nutrients in dry feed with a chemical preservative for «winter conditions», which are characterized by low temperature. It is seen from the results of studies that the rate of hydrolysis and the depth depends on the temperature and the antioxidant origin: enzymatic catalysis accelerated at 29.8 °C («summer» conditions), reducing the temperature (12 °C) slows down the hydrolysis process (the period of «autumn – spring»), but even at 4.5 ° with the enzymatic activity of microbial lipases shown, but to a lesser degree («winter»).Thus, the main causes of spoilage of feed – high temperature (about 30 °C) and high humidity (60 –70%) to facilitate activation of oxidative processes and the development of the microflora, and the duration of storage, even under favorable conditions, depending on the origin of a preservative. Therefore, it is recommended to store this food in a dry, cool place, preferably dark place.

The Impact of Rendered Protein Meal Oxidation Level on Shelf-Life, Sensory Characteristics, and Acceptability in Extruded Pet Food

Simple SummarySensory analysis was used to determine the changes due to the storage time on extruded pet food prepared from two different rendered protein meals: (i) beef meat and bone meal (BMBM); (ii) chicken byproduct meal (CPBM). Extrusion is a process where feed is pressed through a die in order to create shapes and increase digestibility. Descriptive sensory analysis using a human panel found an increase in undesirable sensory attributes (e.g., oxidized oil, rancid) in extruded pet food over storage time, especially the one prepared from chicken by product meal without antioxidants. The small increase in oxidized and rancid aromas of BMBM samples did not affect pet owners’ acceptability of the products. CPBM samples without antioxidants showed a notable increase in oxidized and rancid aroma over storage time and, thus, affected product acceptability negatively. This finding indicated that human sensory analysis can be used as a tool to track the changes of pet food characteristics due to storage, as well as estimate the shelf-life of the products.Pet foods are expected to have a shelf-life for 12 months or more. Sensory analysis can be used to determine changes in products and to estimate products’ shelf-life. The objectives of this study were to (1) investigate how increasing levels of oxidation in rendered protein meals used to produce extruded pet food affected the sensory properties and (2) determine the effect of shelf-life on pet owners’ acceptability of extruded pet food diet formulated without the use of preservative. Pet food diets contained beef meat bone meal (BMBM) and chicken byproduct meal (CBPM) in which the oxidation was retarded with ethoxyquin, mixed tocopherols, or none at all, and then extruded into dry pet foods. These samples represented low, medium, and high oxidation levels, respectively. Samples were stored for 0, 3, 6, 9, and 12 months at ambient temperature. Each time point, samples were evaluated by six highly trained descriptive panelists for sensory attributes related to oxidation. Samples without preservatives were chosen for the acceptability test, since the differences in sensory characteristics over storage time were more distinguishable in those samples. Pet owners evaluated samples for aroma, appearance and overall liking. Descriptive sensory analysis detected significant changes in oxidized-related sensory characteristics over storage time. However, the differences for CBPM samples were more pronounced and directional. The consumer study showed no differences in pet owners’ acceptability for BMBM samples. However, the noticeable increase in aroma characteristics (rancid aroma 0.33–4.21) in CBPM samples over storage time did have a negative effect on consumer’s liking (overall liking 5.52–4.95).

Bacteriophages safely reduce Salmonella contamination in pet food and raw pet food ingredients.

Contamination of pet food with Salmonella is a serious public health concern, and several disease outbreaks have recently occurred due to human exposure to Salmonella tainted pet food. The problem is especially challenging for raw pet foods (which include raw meats, seafood, fruits, and vegetables). These foods are becoming increasingly popular because of their nutritional qualities, but they are also more difficult to maintain Salmonella-free because they lack heat-treatment. Among various methods examined to improve the safety of pet foods (including raw pet food), one intriguing approach is to use bacteriophages to specifically kill Salmonella serotypes. At least 2 phage preparations (SalmoFresh® and Salmonelex™) targeting Salmonella are already FDA cleared for commercial applications to improve the safety of human foods. However, similar preparations are not yet available for pet food applications. Here, we report the results of evaluating one such preparation (SalmoLyse®) in reducing Salmonella levels in various raw pet food ingredients (chicken, tuna, turkey, cantaloupe, and lettuce). Application of SalmoLyse® in low (ca. 2-4×106 PFU/g) and standard (ca. 9×106 PFU/g) concentrations significantly (P < 0.01) reduced (by 60-92%) Salmonella contamination in all raw foods examined compared to control treatments. When SalmoLyse®-treated (ca. 2×107 PFU/g) dry pet food was fed to cats and dogs, it did not trigger any deleterious side effects in the pets. Our data suggest that the bacteriophage cocktail lytic for Salmonella can significantly and safely reduce Salmonella contamination in various raw pet food ingredients.

Modeling the long-term kinetics of Salmonella survival on dry pet food

Due to multiple outbreaks and large-scale product recalls, Salmonella has emerged as a priority pathogen in dry pet food and treats. However, little data are available to quantify risks posed by these classes of products to both pets and their owners. Specifically, the kinetics of Salmonella survival on complex pet food matrices are not available. This study measured the long-term kinetics of Salmonella survival on a dry pet food under storage conditions commonly encountered during production, retail, and in households (aw < 0.60, 23 °C). A Salmonella enterica cocktail of 12 strains isolated from dry pet foods and treats was used to inoculate commercial dry dog food. Salmonella was enumerated on non-selective (BHI) and selective (XLD and BS) media. Results at 570 days indicated an initial relatively rapid decline (up to 54 days), followed by a much slower extended decline phase. The Weibull model provided a satisfactory fit for time series of Log-transformed Salmonella counts from all three media (δ: mean 4.65 day/Log (CFU/g); p: mean 0.364 on BHI). This study provides a survival model that can be applied in quantitative risk assessment models.

An investigation of selected chemical contaminants in commercial pet foods in Egypt.

Our study aimed to identify the levels of various contaminants in both wet and dry commercial pet foods in Egypt. A total of 20 local and imported pet food products (3 samples each) were screened for heavy metals by atomic absorption spectroscopy, for mycotoxins by enzyme-linked immunosorbent assay, and for nitrate and nitrite levels by nitrate-nitrite spectrophotometry. Cat food, on average, had greater concentrations of the metals cadmium, chromium, lead, and tin than dog food. Of the investigated metals, only tin concentration exceeded the safe level compared with the standards of the National Research Council and the European Commission for the dog and cat. According to the guidelines of the Association of American Feed Control Officials for canned pet foods, the nitrate and nitrite contents of examined foods greatly exceeded the recommended level. No total aflatoxins were detected in the surveyed samples. None of the samples analyzed had levels above international limits established by the Food and Agriculture Organization (FAO) of the United Nations for ochratoxin, and only 1 sample exceeded the level for aflatoxin B1. Of the 20 samples analyzed for zearalenone, 4 samples had higher levels than the FAO maximum tolerable levels. These results indicate that pet foods marketed in Egypt, especially cat foods, occasionally contain contaminants that could result in adverse effects in pets.

Validation of pathogen destruction in dry pet foods after production and bagging under simulated industry conditions

Validation of pathogen destruction in dry pet foods after production and bagging under simulated industry conditions

Quantitative assessment of human and pet exposure to Salmonella associated with dry pet foods

Recent Salmonella outbreaks associated with dry pet foods and treats highlight the importance of these foods as previously overlooked exposure vehicles for both pets and humans. In the last decade efforts have been made to raise the safety of this class of products, for instance by upgrading production equipment, cleaning protocols, and finished product testing. However, no comprehensive or quantitative risk profile is available for pet foods, thus limiting the ability to establish safety standards and assess the effectiveness of current and proposed Salmonella control measures. This study sought to develop an ingredients-to-consumer quantitative microbial exposure assessment model to: 1) estimate pet and human exposure to Salmonella via dry pet food, and 2) assess the impact of industry and household-level mitigation strategies on exposure. Data on prevalence and concentration of Salmonella in pet food ingredients, production process parameters, bacterial ecology, and contact transfer in the household were obtained through literature review, industry data, and targeted research. A probabilistic Monte Carlo modeling framework was developed to simulate the production process and basic household exposure routes. Under the range of assumptions adopted in this model, human exposure due to handling pet food is null to minimal if contamination occurs exclusively before extrusion. Exposure increases considerably if recontamination occurs post-extrusion during coating with fat, although mean ingested doses remain modest even at high fat contamination levels, due to the low percent of fat in the finished product. Exposure is highly variable, with the distribution of doses ingested by adult pet owners spanning 3Log CFU per exposure event. Child exposure due to ingestion of 1g of pet food leads to significantly higher doses than adult doses associated with handling the food. Recontamination after extrusion and coating, e.g., via dust or equipment surfaces, may also lead to exposure due to the absence of pathogen reduction steps after extrusion or at consumer households. Exposure is potentially highest when Salmonella is transferred to human food that is left at growth-promoting conditions. This model can be applied to evaluate the impact of alternative Salmonella control measures during production, risk communication to consumers, and regulatory standards.

Evaluation of Rapid Molecular Detection Assays for Salmonella in Challenging Food Matrices at Low Inoculation Levels and Using Difficult-to-Detect Strains

Assays for detection of foodborne pathogens are generally initially evaluated for performance in validation studies carried out according to guidelines provided by validation schemes (e.g., AOAC International or the International Organization for Standardization). End users often perform additional validation studies to evaluate the performance of assays in specific matrices (e.g., specific foods or raw material streams of interest) and with specific pathogen strains. However, these types of end-user validations are typically not well defined. This study was conducted to evaluate a secondary end user validation of four AOAC-validated commercial rapid detection assays (an isothermal nucleic acid amplification, an immunoassay, and two PCR-based assays) for their ability to detect Salmonella in two challenging matrices (dry pet food and dark chocolate). Inclusivity was evaluated with 68 diverse Salmonella strains at low population levels representing the limit of detection (LOD) for each assay. One assay detected all strains at the LOD, two assays detected multiple strains only at 10 times the LOD, and the fourth assay failed to detect two strains (Salmonella bongori and S. enterica subsp. houtenae) even at 1,000 times the LOD; this assay was not further evaluated. The three remaining assays were subsequently evaluated for their ability to detect five selected Salmonella strains in food samples contaminated at fractional levels. Unpaired comparisons revealed no significant difference between the results for each given assay and the results obtained with the reference assay. However, analysis of paired culture-confirmed results revealed assay false-negative rates of 4 to 26% for dry pet food and 12 to 16% for dark chocolate. Overall, our data indicate that rapid assays may have high false-negative rates when performance is evaluated under challenging conditions, including low-moisture matrices, strains that are difficult to detect, injured cells, and low inoculum levels.

Cytotoxic effects of oxytetracycline residues in the bones of broiler chickens following therapeutic oral administration of a water formulation

Tetracyclines, which represent one of the most commonly used antibiotics for poultry, are known to be deposited in bones, where they can remain, despite the observation of appropriate withdrawal times. The aim of the study was to determine the concentration of oxytretracycline (OTC) residues in the bone and muscle of chickens, following the oral administration of a commercially available liquid formulation, and to test their cytotoxic effects on an in vitro cell culture model. Seventy-two 1-day-old broiler chickens were randomly allotted into 2 groups (control and treated animals). OTC (40 mg/kg BW) was administered via drinking water during the 1 to 5 and 20 to 25 days of life periods. At the end of the trial, the birds were slaughtered and the OTC residues in the target tissues were measured by means of liquid chromatography (LC) - tandem mass spectrometry (MS/MS). Cytotoxicity was assessed by evaluating the pro-apoptotic effect of the bone residues on the K562 erythroleukemic line and on the peripheral blood mononuclear cells (PBMC). In all the animals, the OTC residues in the muscle were far below the established MRL of 100 μg/kg. The OTC levels in the bones of the treated animals were instead found in the parts per million (ppm) range. Cell cytotoxicity was assessed by evaluating the pro-apoptotic effect of OTC bone residues on the haematopoietic cell system. This in vitro system has revealed a significant pro-apoptotic effect on both the K562 cell line and PBMC cultures. This result suggests potential human and animal health risks due to the entry of tetracycline residues contained in the bones of treated livestock into the food-chain. This could be of concern, particularly for canine and feline diets, as meat, bone meal, and poultry by-products represent some of the main ingredients of pet foods, especially in the case of dry pet food. Further studies are needed to define the underlying mechanisms of cytotoxicity and to evaluate the in vivo toxicological implications due to the observed in vitro effects.

Validation of the NeoFilm for Yeast and Mold Method for Enumeration of Yeasts and Molds in Select Foods.

NeoFilm Yeast and Mold (Y&M), also known as Sanita-kun Yeasts and Molds, is a simple, effective device used for the enumeration of yeasts and molds. It consists of a nonwoven fabric on which a layer of microbial nutrients is deposited in a film. A 1 mL sample homogenate is applied to the membrane and this, in turn, is incubated for 48-72 h at 25°C. Sample homogenates were prepared using two different diluents for customer convenience: phosphate buffered saline (PBS) and 0.1% peptone water. In comparative testing of breaded chicken nuggets, dry pet food, orange juice concentrate, yogurt, and cake mix, there were statistically significant differences in the counts obtained by the NeoFilm Y&M and U.S. Food and Drug Administration Bacteriological Analytical Manual reference culture methods only in the following instances: medium level for orange juice with PBS as diluent and low level for pet food with 0.1% peptone water as diluent, where reference method counts were higher than those of NeoFilm; medium level for cake mix with PBS, and low and medium levels for cake mix with 0.1% peptone water, where NeoFilm produced higher counts than the reference method. In addition to the method comparison study with five matrixes, robustness and stability/lot-to-lot testing were also performed. Results of robustness testing showed no significant effect on results even with perturbation to three assay parameters simultaneously. Results of testing of three lots of devices ranging in age from 2 to 26 months post-manufacture showed no significant differences in performance.

Evaluating Pediococcus acidilactici and Enterococcus faecium NRRL B-2354 as Thermal Surrogate Microorganisms for Salmonella for In-Plant Validation Studies of Low-Moisture Pet Food Products

Pediococcus acidilactici ATCC 8042 and Enterococcus faecium NRRL B-2354 were investigated as potential surrogates for Salmonella serovars using thermal death time kinetics in products such as dry pet foods. The D-values of P. acidilactici ATCC 8042, E. faecium NRRL B-2354, and a cocktail of seven Salmonella serovars associated with low-moisture products were determined in a preservative-free dry pet food product at moisture levels of 9.1, 17.9, and 27.0% and heated between 76.7 and 87.8°C. The D-values were calculated by least squares linear regression. The D-values of P. acidilactici ATCC 8042 were higher than those for the Salmonella serovar cocktail but lower than those for E. faecium NRRL 2354. At 9.1% moisture, D-values of 6.54, 11.51, and 11.66 min at 76.7°C, 2.66, 3.22, and 4.08 min at 82.2°C, and 1.07, 1.29, and 1.69 min at 87.8°C were calculated for Salmonella serovars, P. acidilactici ATCC 8042, and E. faecium NRRL B-2354, respectively. The data suggest that the thermal inactivation characteristics of P. acidilactici ATCC 8042 can be utilized as a surrogate to predict the response of Salmonella in dry pet food products that are thermally processed at <90°C.

Transmission of Bacterial Zoonotic Pathogens between Pets and Humans: The Role of Pet Food

Recent Salmonella outbreaks associated with dry pet food and treats raised the level of concern for these products as vehicle of pathogen exposure for both pets and their owners. The need to characterize the microbiological and risk profiles of this class of products is currently not supported by sufficient specific data. This systematic review summarizes existing data on the main variables needed to support an ingredients-to-consumer quantitative risk model to (1) describe the microbial ecology of bacterial pathogens in the dry pet food production chain, (2) estimate pet exposure to pathogens through dry food consumption, and (3) assess human exposure and illness incidence due to contact with pet food and pets in the household. Risk models populated with the data here summarized will provide a tool to quantitatively address the emerging public health concerns associated with pet food and the effectiveness of mitigation measures. Results of such models can provide a basis for improvements in production processes, risk communication to consumers, and regulatory action.

Development and inter-laboratory study of a method for quantification of fumonisin B1, B2 and B3 in pet foods

An analytical method to determine fumonisin B1 (FB1), B2 (FB2) and B3 (FB3) in pet foods using a liquid chromatography-electrospray ionization-mass spectrometer (LC-ESI-MS) was developed, and an inter-laboratory study was conducted in eleven laboratories. FB1, FB2 and FB3 were extracted with aqueous acetonitrile. The extract was purified by a multifunctional column, MultiSep 211 Fum, and analysed by LC-ESI-MS. The limit of quantification of fumonisins was estimated to be 0.2 mg/kg for dry and semi-dry pet foods, and 0.1 mg/kg for wet pet food. The calibration curve showed linearity in the range of 0.1-5 ng of fumonisins (0.02-1.0 μg/ml). The values of the mean recovery for FB1 at 0.1-1.0 mg/kg were 93.3-107% and of the relative standard deviation less than 7.9%. These values were 87.3-102 and 8.6% for FB2 and 90.8-102 and 8.6% for FB3, respectively. The mean recovery, repeatability, reproducibility and the Horwitz ratio for FB1 from the inter-laboratory validation study were 92.9-98.9%, 2.6-4.6%, 6.8-10% and 0.41-0.54, respectively. The values for FB2 were 91.5-94.7%, 2.7-5.9%, 6.8-8.9% and 0.33-0.55, respectively, and the values for FB3 were 90.1-94.3%, 3.3-5.9%, 7.3-9.5% and 0.44-0.57, respectively.

Accelerated Solvent Extraction Improves Efficiency of Lipid Removal from Dry Pet Food While Limiting Lipid Oxidation

AbstractAccelerated solvent extraction (ASE) was evaluated for extracting lipids from baked and extruded dry pet foods to determine factors controlling extraction efficiency and effects on lipid oxidation. Hydroperoxide decomposition and new lipid oxidation were minimal at 40 °C but increased at higher extraction temperatures without increasing yields. Maximum extraction required grinding samples to 250 µm particles, presence of polar solvents [chloroform, chloroform/methanol 2:1 (v/v) mixed, hexane/methanol 2:1 (v/v)], and a minimum of 20 min total static extraction time in repeat extraction cycles. Hexane and methanol injected into extraction cells simultaneously but separately was able to nearly duplicate extractions of chloroform/methanol, providing an option for replacing toxic chlorinated hydrocarbon solvents in ASE. However, lipid oxidation was higher in hexane. Yields were quantitative in baked biscuits but lower in extruded kibbles due to more dense, complex molecular structures. ASE extraction yields of 40 min or less were comparable to manual extraction yields of 24–48 h, with lower oxidation. Overall, one or two ASE extraction cycles with static times less than 20 min appeared to provide adequate lipid yields that accurately reflect lipid composition while inducing minimal modification when lipid oxidation products are the analytical endpoint.

DNAble(®) Molecular Detection Kit for Salmonella.

The DNAble Salmonella detection assay utilizes single overnight culture enrichment, user-friendly sample preparation, and isothermal DNA amplification for Salmonella detection. This report describes studies performed in support of AOAC Research Institute Performance Tested Method(SM) certification of the DNAble assay. Selectivity (inclusivity and exclusivity) studies were performed in the sponsor's laboratory. DNAble detected 119 out of 120 Salmonella isolates, representing 100 Salmonella serovars, in the inclusivity study while none of the 35 diverse non-Salmonella strains (32 species) tested was detected in the exclusivity study. Consistency (lot-to-lot and stability), instrument variation, and robustness studies were also conducted by the sponsor. Statistically equivalent assay performance was observed in these studies demonstrating robust assay manufacture and performance despite variation of multiple parameters in these challenges. Matrix studies, performed in an independent laboratory, evaluated DNAble assay performance in dry pet food, on stainless steel surfaces, and poultry environmental drag swab samples. Two sample sizes (25 and 375 g) and two culture volumes (9:1 and 3:1, v/w) were evaluated in separate matrix studies for dry pet food to provide multiple certified testing options for assay users. DNAble assay performance for dry pet food and stainless steel was compared to the procedures described in the U.S. Food and Drug Administration (FDA) Bacteriological Analytical Manual, Chapter 5, Salmonella. Assay performance for drag swabs was compared to protocols dictated in the FDA Environmental Sampling and Detection of Salmonella in Poultry Houses guidelines. Matrix study results demonstrated statistically equivalent DNAble assay performance compared to these reference methods, ensuring that the DNAble assay provides results comparable to those of the reference methods.

Validation of the Soleris direct yeast and mold method for semiquantitative determination of yeast and mold in a variety of foods.

A study was carried out to determine the efficacy of the Soleris Direct Yeast and Mold (DYM) automated growth-based method for semiquantitative detection of yeast and mold in a variety of food products. A probability of detection (POD) statistical model was used to compare Soleris results at multiple test thresholds (dilutions) with plate counts determined using the U.S. Food and Drug Administration Bacteriological Analytical Manual, Chapter 18, dilution plating procedure. Fourteen naturally contaminated food products were tested, with Soleris testing performed at three or more threshold levels for each food. Using the POD model, the majority of Soleris test results were in statistical agreement with the reference plating procedures. The exceptions included a single threshold level in yogurt, black pepper, dried fruit, and dry pet food, and two levels in nonfat dry milk and saw palmetto powder. In all but one of these instances, the exception being pet food, the statistical disagreement was due to Soleris estimating a higher level of contamination than the reference method. Results of ruggedness testing showed that the Soleris method produced accurate results even when significant variances in a critical operating parameter, incubation temperature, were introduced. Results of the internal and independent laboratory validation studies showed that the Soleris DYM method can be used as an accurate alternative to conventional dilution plating procedures for evaluation of yeast and mold counts at threshold levels, while saving as much as 72 h in analysis time.