Dry Pet Food Research Articles

Determination of moulds and mycotoxins in dry dog and cat food using liquid chromatography with mass spectrometry and fluorescence detection

In this study moulds and 12 mycotoxins in dry pet food samples (25 for dogs and 24 for cats) were determined. Primary moulds identified were Aspergillus, Mucor and Penicillium, found in 55% of the samples. Deoxynivalenol and zearalenone (ZEN) were detected in all samples with mean respective concentrations being 97.3 and 38.3 µg kg−1 in cat food and 114 and 20.1 µg kg−1 in dog food. T-2 and HT-2 toxins were present in 88% and 84% of the samples, respectively. Two samples contained fumonisins, with a maximum concentration of 108 µg kg−1. Aflatoxin B1 and ochratoxin A were detected in 8% and 45% of the samples, respectively. The measured mould and mycotoxin levels were consistent with results obtained by other studies. However, potential exposure to relatively high concentrations of an oestrogen mycotoxin as is ZEN, especially when in combination with other mycotoxins, needs attention.

Outbreak of Salmonella enterica serotype Infantis infection in humans linked to dry dog food in the United States and Canada, 2012.

CASE DESCRIPTION--In April 2012, Salmonella enterica serotype Infantis was detected in an unopened bag of dry dog food collected during routine retail surveillance. PulseNet, a national bacterial subtyping network, identified humans with Salmonella Infantis infection with the same genetic fingerprint as the dog food sample. CLINICAL FINDINGS--An outbreak investigation identified 53 ill humans infected with the outbreak strain during January 1 to July 5, 2012, in 21 states and 2 provinces in Canada; 20 (38%) were children ≤ 2 years old, and 12 of 37 (32%) were hospitalized. Of 21 ill people who remembered the dog food brand, 12 (57%) reported a brand produced at a plant in Gaston, SC. Traceback investigations also identified that plant. The outbreak strain was isolated from bags of dry dog food and fecal specimens obtained from dogs that lived with ill people and that ate the implicated dry dog food. TREATMENT AND OUTCOME--The plant was closed temporarily for cleaning and disinfection. Sixteen brands involving > 27,000 metric tons (> 30,000 tons) of dry dog and cat food were recalled. Thirty-one ill dogs linked to recalled products were reported through the FDA consumer complaint system. CLINICAL RELEVANCE-- A one-health collaborative effort on epidemiological, laboratory, and traceback investigations linked dry dog foods produced at a plant to illnesses in dogs and humans. More efforts are needed to increase awareness among pet owners, health-care professionals, and the pet food industry on the risk of illness in pets and their owners associated with dry pet foods and treats.

Validation of the ANSR Salmonella method for detection of Salmonella spp. in a variety of foods. Performance Tested Method 061203.

This study represents a proposal to extend the matrix claims for the ANSR Salmonella test, Performance Tested Method 061203. The test is based on the nicking enzyme amplification reaction (NEAR) isothermal nucleic acid amplification technology. The assay platform features simple instrumentation, minimal labor, and following a single-step 16-24 h enrichment (depending on sample type), an extremely short assay time of 30 min including sample preparation. Detection is real-time using fluorescent molecular beacon probes. ANSR Salmonella was originally validated for detection of Salmonella spp. in chicken carcass rinse, raw ground turkey, raw ground beef, hot dogs, and oat cereal, and on stainless steel, plastic, sealed concrete, ceramic tile, and rubber surfaces. The matrixes tested in this study include pet food, ice cream, soy flour, raw almonds, peanut butter, spinach, black pepper, raw frozen shrimp, cocoa powder, and pasteurized dried egg. In unpaired comparative testing there were no statistically significant differences in the number of positive results obtained with the ANSR and the reference culture methods. Enrichment for 16 h was effective for all commodities tested except ice cream, black pepper, dried pasteurized egg, and 375 g samples of dry pet food, for which enrichment for 24 h is indicated.

Pet Food Processing—Understanding Transformations in Starch during Extrusion and Baking

Pet foods contain a multitude of ingredients as they must satisfy all of a pet's daily nutritional requirements, including carbohydrates, protein, fat, and micronutrients. Processing dry pet foods typically involves baking or extrusion. Extrusion has a number of advantages over conventional batch methods: a single extruder can perform several functions, and a wide variety of products can be processed using the same equipment by altering the processing conditions, screw profile, die, and ingredients. Due to the nature of the baking process, baking is a much lower throughput process compared with extrusion. Starch is an important part of a typical dry pet food formulation and undergoes several important changes during processing that impact the digestibility, palatability, and physical attributes of the final product. Notable starch-related phenomena include gelatinization and amylose-lipid complexation. Proximate analysis of dry pet foods after baking or extrusion confirmed that final products were isonutr...

Rapid detection of Salmonella in pet food: design and evaluation of integrated methods based on real-time PCR detection.

Reducing the risk of Salmonella contamination in pet food is critical for both companion animals and humans, and its importance is reflected by the substantial increase in the demand for pathogen testing. Accurate and rapid detection of foodborne pathogens improves food safety, protects the public health, and benefits food producers by assuring product quality while facilitating product release in a timely manner. Traditional culture-based methods for Salmonella screening are laborious and can take 5 to 7 days to obtain definitive results. In this study, we developed two methods for the detection of low levels of Salmonella in pet food using real-time PCR: (i) detection of Salmonella in 25 g of dried pet food in less than 14 h with an automated magnetic bead-based nucleic acid extraction method and (ii) detection of Salmonella in 375 g of composite dry pet food matrix in less than 24 h with a manual centrifugation-based nucleic acid preparation method. Both methods included a preclarification step using a novel protocol that removes food matrix-associated debris and PCR inhibitors and improves the sensitivity of detection. Validation studies revealed no significant differences between the two real-time PCR methods and the standard U.S. Food and Drug Administration Bacteriological Analytical Manual (chapter 5) culture confirmation method.

The impact of home-prepared diets and home oral hygiene on oral health in cats and dogs

Many factors influence the oral health status of cats and dogs. The present study aimed to elucidate the influence of feeding home-prepared (HP) food v. commercial pet food on oral health parameters in these animals and to investigate the effect of home oral hygiene on oral health. The study surveyed 17,184 dogs and 6371 cats visiting over 700 Polish veterinary surgeries in 2006-7 during a Pet Smile activity organised by the Polish Small Animal Veterinary Association. All animals underwent conscious examinations to assess dental deposits, size of mandibular lymph nodes and gingival health. An oral health index (OHI) ranging from 0 to 8 was calculated for each animal by combining examination scores, where 0 indicates good oral health and 8 indicates poorest oral health. Information was collected on age, diet and home oral hygiene regimens. There was a significant effect of diet on the OHI (P < 0.001) whereby feeding the HP diet increased the probability of an oral health problem in both cats and dogs. There was a significant beneficial effect of feeding only commercial pet food compared with the HP diet when at least part of the diet was composed of dry pet food. Daily tooth brushing or the offering of daily dental treats were both effective in significantly reducing the OHI in both cats and dogs compared with those receiving sporadic or no home oral hygiene. Feeding only a dry diet was beneficial for oral health in cats and dogs. Tooth brushing and the offering of dental treats were very effective in maintaining oral health, provided they were practised daily.

Reveal® Salmonella 2.0 Test for Detection of Salmonella spp. in Foods and Environmental Samples

Reveal Salmonella 2.0 is an improved version of the original Reveal Salmonella lateral flow immunoassay and is applicable to the detection of Salmonella enterica serogroups A-E in a variety of food and environmental samples. A Performance Tested Method validation study was conducted to compare performance of the Reveal 2.0 method with that of the U.S. Department of Agriculture-Food Safety and Inspection Service or U.S. Food and Drug Administration/Bacteriological Analytical Manual reference culture methods for detection of Salmonella spp. in chicken carcass rinse, raw ground turkey, raw ground beef, hot dogs, raw shrimp, a ready-to-eat meal product, dry pet food, ice cream, spinach, cantaloupe, peanut butter, stainless steel surface, and sprout irrigation water. In a total of 17 trials performed internally and four trials performed in an independent laboratory, there were no statistically significant differences in performance of the Reveal 2.0 and reference culture procedures as determined by Chi-square analysis, with the exception of one trial with stainless steel surface and one trial with sprout irrigation water where there were significantly more positive results by the Reveal 2.0 method. Considering all data generated in testing food samples using enrichment procedures specifically designed for the Reveal method, overall sensitivity of the Reveal method relative to the reference culture methods was 99%. In testing environmental samples, sensitivity of the Reveal method relative to the reference culture method was 164%. For select foods, use of the Reveal test in conjunction with reference method enrichment resulted in overall sensitivity of 92%. There were no unconfirmed positive results on uninoculated control samples in any trials for specificity of 100%. In inclusivity testing, 102 different Salmonella serovars belonging to serogroups A-E were tested and 99 were consistently positive in the Reveal test. In exclusivity testing of 33 strains of non-salmonellae representing 14 genera, 32 were negative when tested with Reveal following nonselective enrichment, and the remaining strain was found to be substantially inhibited by the enrichment media used with the Reveal method. Results of ruggedness testing showed that the Reveal test produces accurate results even with substantial deviation in sample volume or device development time.

Reveal®Salmonella 2.0 Test for Detection of Salmonella spp. in Foods and Environmental Samples

Abstract Reveal Salmonella 2.0 is an improved version of the original Reveal Salmonella lateral flow immunoassay and is applicable to the detection of Salmonella enterica serogroups A–E in a variety of food and environmental samples. A Performance Tested MethodSM validation study was conducted to compare performance of the Reveal 2.0 method with that of the U.S. Department of Agriculture-Food Safety and Inspection Service or U.S. Food and Drug Administration/Bacteriological Analytical Manual reference culture methods for detection of Salmonella spp. in chicken carcass rinse, raw ground turkey, raw ground beef, hot dogs, raw shrimp, a ready-to-eat meal product, dry pet food, ice cream, spinach, cantaloupe, peanut butter, stainless steel surface, and sprout irrigation water. In a total of 17 trials performed internally and four trials performed in an independent laboratory, there were no statistically significant differences in performance of the Reveal 2.0 and reference culture procedures as determined by Chi-square analysis, with the exception of one trial with stainless steel surface and one trial with sprout irrigation water where there were significantly more positive results by the Reveal 2.0 method. Considering all data generated in testing food samples using enrichment procedures specifically designed for the Reveal method, overall sensitivity of the Reveal method relative to the reference culture methods was 99%. In testing environmental samples, sensitivity of the Reveal method relative to the reference culture method was 164%. For select foods, use of the Reveal test in conjunction with reference method enrichment resulted in overall sensitivity of 92%. There were no unconfirmed positive results on uninoculated control samples in any trials for specificity of 100%. In inclusivity testing, 102 different Salmonella serovars belonging to serogroups A–E were tested and 99 were consistently positive in the Reveal test. In exclusivity testing of 33 strains of non-salmonellae representing 14 genera, 32 were negative when tested with Reveal following nonselective enrichment, and the remaining strain was found to be substantially inhibited by the enrichment media used with the Reveal method. Results of ruggedness testing showed that the Reveal test produces accurate results even with substantial deviation in sample volume or device development time.

Evaluation of Applied Biosystems MicroSEQ® Real-Time PCR System for Detection of Salmonella spp. in Food

Real-time PCR methods for detecting foodborne pathogens offer the advantages of simplicity and quick time-to-results compared to traditional culture methods. In this study, the MicroSEQ real-time PCR system was evaluated for detection of Salmonella spp. in 10 different food matrixes following the AOAC Research Institute's Performance Tested Method validation program. In addition, the performance of the MicroSEQ system was evaluated for the detection of Salmonella in peanut butter as a part of the Emergency Response Validation Program sponsored by the AOAC Research Institute. The system was compared to the ISO 6579 reference method using a paired-study design for detecting Salmonella spp. in raw ground beef, raw chicken, raw shrimp, Brie cheese, shell eggs, cantaloupe, chocolate, black pepper, dry infant formula, and dry pet food. For the peanut butter study, the system was compared to the U.S. Food and Drug Administration's Bacteriological Analytical Manual procedures using an unpaired-study design. No significant difference in performance was observed between the MicroSEQ Salmonella spp. detection system and the corresponding reference methods for all 11 food matrixes. The MicroSEQ system detected all Salmonella strains tested, while showing good discrimination against detection of an exclusivity panel of 30 strains, with high accuracy.

House dust and storage mite contamination of dry dog food stored in open bags and sealed boxes in 10 domestic households

Dry pet food is a potential source of exposure to house dust and storage mite allergens in canine atopic dermatitis. This study evaluated contamination of house dust and dry dog food stored in paper bags, sealable plastic bags and sealable plastic boxes in 10 households for 90 days using Acarex(®) tests for guanine, a Der p 1 ELISA and mite flotation. Acarex(®) tests were negative in all the food samples but positive in all the house dust samples. The Der p 1 levels and mite numbers significantly increased in food from paper bags (P = 0.0073 and P = 0.02, respectively), but not plastic bags or boxes. Mite numbers and Der p 1 levels were 10-1000 times higher in house dust than the corresponding food samples (P < 0.0001). There were significant correlations between Der p 1 in house dust and food from the paper (P < 0.0001) and plastic bags (P = 0.003), and mite numbers in house dust and food from the paper bags (P = 0.0007). Bedding and carpets were significantly associated with Der p 1 levels in house dust (P = 0.015 and P = 0.01, respectively), and food from the paper (both P = 0.02) and plastic bags (P = 0.03 and P = 0.04, respectively). Mites were identified in six of 10 paper bag, three of 10 plastic bag, one of 10 plastic box and nine of 10 house dust samples. These comprised Dermatophagoides (54%), Tyrophagus (10%; all from food) and unidentified mites (36%). Storage of food in sealable plastic boxes largely prevented contamination for 3 months. Exposure to mites and mite proteins in all the stored food, however, appeared to be trivial compared with house dust.

Human Salmonella Infections Linked to Contaminated Dry Dog and Cat Food, 2006–2008

Human Salmonella infections associated with dry pet food have not been previously reported. We investigated such an outbreak of Salmonella Schwarzengrund and primarily affecting young children. Two multistate case-control studies were conducted to determine the source and mode of infections among case-patients with the outbreak strain. Study 1 evaluated household exposures to animals and pet foods, and study 2 examined risk factors for transmission among infant case-patients. Environmental investigations were conducted. Seventy-nine case-patients in 21 states were identified; 48% were children aged 2 years or younger. Case-households were significantly more likely than control households to report dog contact (matched odds ratio [mOR]: 3.6) and to have recently purchased manufacturer X brands of dry pet food (mOR: 6.9). Illness among infant case-patients was significantly associated with feeding pets in the kitchen (OR: 4.4). The outbreak strain was isolated from opened bags of dry dog food produced at plant X, fecal specimens from dogs that ate manufacturer X dry dog food, and an environmental sample and unopened bags of dog and cat foods from plant X. More than 23 000 tons of pet foods were recalled. After additional outbreak-linked illnesses were identified during 2008, the company recalled 105 brands of dry pet food and permanently closed plant X. Dry dog and cat foods manufactured at plant X were linked to human illness for a 3-year period. This outbreak highlights the importance of proper handling and storage of pet foods in the home to prevent human illness, especially among young children.

Multi-Analyst, Multi-Matrix Performance of the QuEChERS Approach for Pesticide Residues in Foods and Feeds Using HPLC/MS/MS Analysis with Different Calibration Techniques

Three different calibration approaches were applied in HPLC/MS/MS using electrospray ionization for the determination of 14 diverse pesticide residues at different levels in a variety of food matrixes. This study was conducted as part of a 4 day training course for 17 chemists to learn the "quick, easy, cheap, effective, rugged, and safe" (QuEChERS) approach to pesticide residue analysis in foods. The analysts were divided into four different teams for the analysis of 12 different matrixes (strawberries, plums, carrots, green peppers, milk, molasses, alfalfa oats, corn silage, dry pet food, soybean, almonds, and foliage). The acetate-buffered QuEChERS protocol gave excellent results in the spiked samples for all matrixes tested. The HPLC/MS/MS calibration techniques consisted of external standardization in solvent solutions, matrix-matching, and the echo-peak technique. Peak areas were normalized to an internal standard in all three approaches. Matrix effects were observed with the corn silage, carrot, and foliage extracts, but they were minimal or nonexistent in the other matrixes. Matrix-matching best compensated for matrix effects, but has logistical difficulties in real-world application and required extra sample preparation compared to the other approaches. The echo-peak technique reduced matrix effects but did not eliminate them.

VIDAS Salmonella (SLM) Assay Method EasySLM with ChromID Salmonella (SM2) Agar

A method modification study was conducted for the VIDAS Salmonella (SLM) assay (AOAC Performance Tested Method 020901) using the EasySLM method to validate a matrix extension for peanut butter. The VIDAS EasySLM method is a simple enrichment procedure compared to traditional Salmonella methods, requiring only pre-enrichment and a single selective enrichment media, Salmonella Xpress 2 (SX2) broth. SX2 replaces the two selective broths in traditional methods and eliminates the M broth transfer, incubation, and subsequent pooling of M broths prior to VIDAS assay. The validation study was conducted under the AOAC Research Institute Emergency Response Validation program. VIDAS SLM was compared to the U.S. Food and Drug Administration's Bacteriological Analytical Manual (FDA-BAM) method for detection of S. enterica ser. Typhimurium in peanut butter. All peanut butter samples were prepared, blind-coded, and shipped to the method developers' laboratory by Q Laboratories. In addition, Q Laboratories performed most probable number and reference method analyses on peanut butter samples. The VIDAS EasySLM ChromID Salmonella (SM2) Agar was previously validated in the Performance Tested Methods program for the detection of Salmonella in roast beef, raw ground pork, turkey, pork sausage, raw chicken breast, dry pet food, whole milk, ice cream, bagged spinach, shrimp (raw, peeled), raw cod, spent irrigation water, pecans, peanut butter, dry pasta, cake mix, ground black pepper, nonfat dry milk, liquid eggs, cantaloupe, and orange juice. In the matrix extension study for peanut butter, the VIDAS EasySLM method was shown to be equivalent to the appropriate reference culture procedure using both buffered peptone water pre-enrichment and the FDA-BAM lactose pre-enrichment in the two-step enrichment method with SX2 media. The current study extends the validation to include peanut butter.

Foodproof Salmonella Detection Kit

The foodproof Salmonella Detection Kit was previously validated in the Performance Tested Methods program for the detection of Salmonella species in a variety of foods, including milk powder, egg powder, coconut, cocoa powder, chicken breast, minced meat, sliced sausage, sausage, smoked fish, pasta, white pepper, cumin, dough, wet pet food, dry pet food, ice cream, watermelon, sliced cabbage, food dye, and milk chocolate. The method was shown to be equivalent to the U.S. Food and Drug Administration's Bacteriological Analytical Manual (FDA-BAM) and the U.S. Department of Agriculture, Food Safety and Inspection Service's Microbiology Laboratory Guidebook reference culture procedures. In the first Emergency Response Validation (ERV) extension study, peanut butter was inoculated with S. enterica. ser Typhimurium. For the low inoculation level (1.08 CFU/25 g), a Chi-square value of 2.25 indicated that there was no significant performance difference between the foodproof Salmonella Detection Kit and the FDA-BAM reference method. For high-level inoculation (11.5 CFU/25 g) and uninoculated control, there was 100% agreement between the methods. In the second ERV extension study, peanut butter was inoculated with S. enterica. ser Typhimurium. For both inoculation levels (0.1 and 0.5 CFU/25 g by most probable number), Chi-square values of 0 indicated that there was no significant performance difference between foodproof Salmonella Detection Kit and the FDA-BAM reference method.

Applicability of Pressurized Liquid Extraction for Melamine Analysis in Pet Foods with High-Performance Liquid Chromatography with Diode Array Detection

A pressurized liquid extraction (PLE) method was developed for melamine analysis in pet foods. The PLE method which utilized an accelerated solvent extraction (ASE®) system was also compared with sonication and polytron extraction methods. The parameters for the optimized PLE method were temperature (75 °C for wet pet food, 125 °C for dry pet food), pressure (1,500 psi), static time (10 min), flush volume (40%), purge time (1 min), and number of cycles (1). Recoveries obtained by PLE method were significantly higher (P ≤ 0.05) than those of sonication and polytron methods for dry pet food samples. For the analysis of adulterated pet foods, PLE resulted in the highest melamine content followed by sonication and polytron. Using PLE, samples fortified with melamine at 2.5 and 100 mg kg−1 resulted in recoveries ranging from 55% to 90% for wet samples and from 90% to 116% for dry samples. Low recovery rate from wet samples at low spike level (2.5 mg kg−1) may have been caused by co-aggregation of polysaccharide and protein with melamine due to low pH during solid-phase extraction cleanup. Limit of detection and limit of quantification values were 0.5 (mg kg−1) and 1.0 (mg kg−1) for dry samples. Overall, PLE had the best extraction efficiency compared to sonication and polytron, proving PLE to be a useful tool for melamine analysis of pet foods.

Ataxia and paralysis in cats in Australia associated with exposure to an imported gamma‐irradiated commercial dry pet food

Between June 2008 and March 2009, 87 cats in Australia developed symmetrical hindlimb ataxia, paraparesis, tetraparesis, paraplegia or tetraplegia in association with eating an imported, irradiated dry pet food. This communication reports the clinical signs and outcomes of those cats.

In this issue – September 2009

In this issue – September 2009

Study of the chemical and nutritional characteristics of commercial dog foods used as elimination diet for the diagnosis of canine food allergy

ReferencesHypoallergenic” pet foods are commercial dietary products for dogs and cats used as elimination diets for the diagnosis of adverse food reactions. Aim of this study was to compare chemical and nutritional characteristics of this kind of dog foods with regular maintenance diets. Twenty-nine dry pet foods (pellets) were collected and divided into classes on the basis of the type (H: hypoallergenic; R: regular), source of fat (with or without fish oil) and source of protein (with or without fish protein) used in their composition. Labels of the H pet foods identified 8 products (44%) with one protein in their formula, suggesting that only few commercial manufacturers concern about the number of protein sources included in their products. Samples of the two groups showed different chemical profiles with lower levels of protein, gross energy, phosphorus and better fatty acid profile (expressed as % of total fatty acids) for H products in comparison to R foods: PUFA, 38.91 vs 24.03, P<0.01; ω3, 5.70 vs 2.58, P<0.01; ω6, 33.22 vs 21.63, P<0.01; DHA, 2.85 vs 0.16, P<0.05; CLA, 0.24 vs 0.08, P<0.05, for H and R respectively. This study suggests that the differences observed in the fatty acids composition may be attributed to fish proteins addition, but not to fish oil, in H pet foods production.

Towards Understanding the Impacts of the Pet Food Industry on World Fish and Seafood Supplies

The status of wild capture fisheries has induced many fisheries and conservation scientists to express concerns about the concept of using forage fish after reduction to fishmeal and fish oil, as feed for farmed animals, particularly in aquaculture. However, a very large quantity of forage fish is being also used untransformed (fresh or frozen) globally for other purposes, such as the pet food industry. So far, no attempts have been made to estimate this quantum, and have been omitted in previous fishmeal and fish oil exploitation surveys. On the basis of recently released data on the Australian importation of fresh or frozen fish for the canned cat food industry, here we show that the estimated amount of raw fishery products directly utilized by the cat food industry equates to 2.48 million metric tonnes per year. This estimate, plus the previously reported global fishmeal consumption for the production of dry pet food suggest that 13.5% of the total 39.0 million tonnes of wild caught forage fish is used for purposes other than human food production. This study attempts to bring forth information on the direct use of fresh or frozen forage fish in the pet food sector that appears to have received little attention to this date and that needs to be considered in the global debate on the ethical nature of current practices on the use of forage fish, a limited biological resource.

Evaluation of storage mite contamination of commercial dry dog food

Storage mites may be considered important allergens in dogs with atopic dermatitis. High sensitization rates to Tyrophagus, Acarus, and Lepidoglyphus species have been reported in atopic dogs, and dry pet food has been suggested as a potential source of storage mite exposure. The aim of the present study was to evaluate commercial dry dog food for contamination with storage mites, and how storage time and conditions could influence the risk of contamination. Ten different premium commercial dry dog foods formulated for skin disorders were selected. Food bags were opened and stored for 6 weeks under two different environmental conditions. At different time points, samples from each bag were collected and analysed by microscopy, guanine test, storage mite-specific traps, and a modified flotation technique. On opening, two storage mites identified as Acarus siro were isolated from one of the 10 bags by flotation technique, indicating that storage mites can be present in packaged dry dog food bags. After 5 weeks of storage under environmental conditions optimal for mite growth (23.2 +/- 2.1 degrees C and 71 +/- 5.6% of relative humidity), mites were detected by microscopic observation in nine of the 10 diets. When mites were identified by the flotation technique, Tyrophagus spp. were found to be the most common contaminating species. These results show that dry dog food can be a suitable substrate for storage mite reproduction, and that environmental and storage conditions may influence food contamination and mite development.