Cereal Samples Research Articles

A Novel Immunoaffinity Column Based Metal-Organic Framework Deep Eutectic Solvents@Molecularly Imprinted Polymers Coupled with High Performance Liquid Chromatography – Fluorescence Detection for the Extraction of Aflatoxins Afb1, Afb2, Afg1 and Afg2 from Cereals Samples of Khuzestan Province of Iran

A Novel Immunoaffinity Column Based Metal-Organic Framework Deep Eutectic Solvents@Molecularly Imprinted Polymers Coupled with High Performance Liquid Chromatography – Fluorescence Detection for the Extraction of Aflatoxins Afb1, Afb2, Afg1 and Afg2 from Cereals Samples of Khuzestan Province of Iran

Multistate outbreak of Salmonella Mbandaka infections linked to sweetened puffed wheat cereal - United States, 2018.

In May of 2018, PulseNet, the national molecular subtyping network for enteric pathogens, detected a multistate cluster of illnesses caused by an uncommon molecular subtype of Salmonella serovar Mbandaka. A case was defined as an illness in a person infected with the outbreak strain of Salmonella Mbandaka with illness onset on or after 3 March 2018 and before 1 September 2018. One-hundred thirty-six cases from 36 states were identified; 35 hospitalisations and no deaths were reported. Ill people ranged in age from <1 year to 95 years (median: 57 years). When standardised questionnaires did not generate a strong hypothesis, opened-ended interviews were performed. Sixty-three of 84 (75%) ultimately reported consuming or possibly consuming a specific sweetened puffed wheat cereal in the week before illness onset. Environmental sampling performed at the cereal manufacturing facility yielded the outbreak strain. The outbreak strain was also isolated from open cereal samples from ill people's homes and from a sealed retail sample. Due to these findings, the brand owner of the product issued a voluntary recall of the cereal on 14 June 2018. Additional investigation of the manufacturing facility identified persistent environmental contamination with Salmonella Mbandaka that was closely genetically related to other isolates in the outbreak. This investigation highlights the ability of Salmonella to survive in low-moisture environments, and the potential for prolonged outbreaks linked to products with long shelf lives and large distribution areas.

Survey for Legume and Cereal Viruses in Libya

Abukraa, H., S.G. Kumari and F. Bshia. 2022. Survey for Legume and Cereal Viruses in Libya. Arab Journal of Plant Protection, 40(3): 222-230. https://doi.org/10.22268/AJPP-40.3.222230 A field survey was conducted during March and April 2010, to identify viral diseases affecting cereal and legume crops in different regions of Libya. A total of 3706 barley and wheat samples were collected randomly in addition to 187 symptomatic samples from 22 barley and 20 wheat fields. Moreover, 34 symptomatic legume samples were collected from two faba bean fields and one field each of of lentil, chickpea, Vicia sativa, Vicia ervillia and Vicia narbonesis. All samples were tested at ICARDA Virology Laboratory by tissue blot immunoassay (TBIA) using specific antibodies. Results of random cereal samples showed that Barley yellow dwarf virus-PAV (BYDVPAV) was more common (12.6%) in wheat and barley samples, followed by Wheat dwarf virus (WDV) (1.1%). In symptomatic samples, BYDV-PAV was also the most common (45 out of 187 samples tested were infected: 45/187), followed by WDV (23/187), Barley yellow striate mosaic virus (BYSMV) (40/187) and Maize streak virus (MSV) (7/187). In legume fields, Soybean dwarf virus (SbDV) 13 out of 34 samples tested were infected (13/34) was the most common, followed by Bean leafroll virus (BLRV) (12/34), Chickpea chlorotic stunt virus (CpCSV) (5/34) and Beet western yellows virus (BWYV) (4/34). Serological results were confirmed by amplification with polymerase chain reaction (PCR) using specific primers. This is the first report of WDV, MSV and BYSMV on cereal crops, and BLRV, BWYV, SbDV and CpCSV on legume crops in Libya. Keywords: Libya, Cereals, Legumes, Viruses, TBIA, PCR

A Novel Immunoaffinity Column Based Metal-Organic Framework Deep Eutectic Solvents@Molecularly Imprinted Polymers Coupled with High Performance Liquid Chromatography – Fluorescence Detection for Extraction of Aflatoxins Afb1, Afb2, Afg1 and Afg2 from Cereals Samples of Khuzestan Province of Iran

A Novel Immunoaffinity Column Based Metal-Organic Framework Deep Eutectic Solvents@Molecularly Imprinted Polymers Coupled with High Performance Liquid Chromatography – Fluorescence Detection for Extraction of Aflatoxins Afb1, Afb2, Afg1 and Afg2 from Cereals Samples of Khuzestan Province of Iran

The content of dietary fiber in cereal-based complementary foods

The article presents analytical data on the content of soluble and insoluble dietary fiber in complementary foods for children from 4 months and for pregnant women. 37 samples of cereals from various manufacturers and 4 samples of porridge for pregnant women were analyzed. These products were developed on the basis of buckwheat, wheat, rice, corn and oat flour, with or without the addition of milk powder, and with or without the addition of flavoring ingredients: apple, peach, banana, apricot, quince, pumpkin. As the results of the study showed, the total content of dietary fiber (DF) in the samples under study varied greatly from 0.6% («Malych» (Istra, rice flour) up to 21.6% («Heinz, Mom's porridge», based on buckwheat flour), which depended on the quantitative composition of complementary foods and the type of flour introduced, as well as on the availability of milk and flavor fillers. In the samples of cereals of the «Malych» series, with the same percentage of bookmarks in the recipe composition of cereal flour, the total content of DF was 0.6, 1.3 and 2.25% for rice, buckwheat and oat flour, respectively. In the samples of cereals of the «Heinz» series, the total DF content was 5.4% based on corn flour, 10.0% with rice, 10.5% with wheat and 16.3% each with buckwheat and oat flour. The addition of fruit additives to the recipe leads to an increase in the DF content in the product from 13.1% («Heinz» buckwheat flour with apple) to 18.6% («Heinz» buckwheat flour with quince). In a series of Heinz porridges based on oatmeal with various flavorings, the results of the study were: without filler - the total content of DF was 16.3%; with the addition of apple - 17.9% and with the addition of apricot and apple - 19.7%. In multi-cereal porridges, the total content of dietary fiber ranged from 4.4% «Porridge of seven cereals» to 10.1% «Milk porridge of three cereals with a banana».

Comparative analysis of mycotoxin content in grains of cereal crops

Purpose. To establish the presence and range of mycotoxins in feed grains widely used in farm animal feed. Methods. The content of mycotoxins was determined in grain samples of cereals (wheat, barley, corn, oats). Enzyme-linked immunosorbent assay using the Sunrise analyzer was used for determination. The method involves the use of test systems (manufactured by “Neogen”), which allow to determine six major mycotoxins: aflatoxin, deoxynivalenone, zearalenone, fumonisin, ochratoxin, T-2 toxin. The basis of this method is the attachment to the antibody enzyme label, which allows taking into account the result of the antigen-antibody reaction on detecting enzymatic activity or changing its level. The antibody is specific for a particular type of mycotoxin. Results. The most harmful of the known mycotoxins are aflatoxin, zearalenone, deoxynivalenone, T-2 toxin, ochratoxin, fuminisin. Determination of the first four was carried out in the feed grain of cereals (wheat, barley, oats, corn). The content of aflatoxin was within the range 0.017-0.21 mg/kg, an acceptable level in accordance with the standards (0.1 mg/kg). The amount of zearalenone ranged from 0.10 mg/kg to 3.51 mg/kg at an acceptable level of 3.0 mg/kg. As to deoxynivalenone, the content of this mycotoxin ranged from 0.52 mg/kg to 3.14 mg/kg at an acceptable level of 3.0 mg/kg. Contamination of feed grains with mycotoxin T-2 toxin is defined in the range 0.02-0.60 mg/kg at its permissible level 0.2 mg/kg. The maximum level of each of the mycotoxins was determined in corn grain, which is most often infected with molds. Currently, the only way to hold in check these hazardous substances is to control their content and make every effort to reduce their amount in feed and raw materials. Conclusion. Analyzing the data of the conducted researches, it can be stated that among all grain cereals corn is the most often affected by molds, so the content of mycotoxins in this feed is the highest. Determination of the content of aflatoxin, zearalenone, deoxynivalenol, T-2 toxin and other mycotoxins in grain and feed is aimed at preventing their negative impact on animals and humans.

Development of a biotinylated nanobody for sensitive detection of aflatoxin B1 in cereal via ELISA

Aflatoxin B1 (AFB1) contamination is a severe threat to food safety and human health, and requires continuous monitoring. In this study, we developed a biotin-streptavidin-amplified enzyme-linked immunosorbent assay (BA-ELISA) by using biotinylated nanobody Nb26 and streptavidin-conjugated polymerized horseradish peroxide (SA-PolyHRP) for sensitive and rapid detection of AFB1 in cereal. Under the optimal condition, the IC50 value of the BA-ELISA was improved to 0.21 ng mL−1 for AFB1, satisfying the requirement of detection limit in practical applications. The total assay time of our strategy is reduced to 50 min from 2 h in conventional competitive ELISA. Additionally, the BA-ELISA saves as much as 98% of the antibody in comparison to the previous classic ELISA. Our work also demonstrated an interesting phenomenon that the biotinylated Nb26 achieved better selectivity to AFB1, which could possibly result from the steric hindrance that interferes reaction between the Nb26 and the AFB1 analogs. Furthermore, the assay was used to detect AFB1 in two cereal samples, and the results were in good agreement with that obtained by high performance liquid chromatography. The developed BA-ELISA can be used for routine screening analysis of AFB1, and offers a promising strategy for measuring low concentrations of food contaminants.

A Novel Colorimetric Nano Aptasensor for Ultrasensitive Detection of Aflatoxin B1 Based on the Exonuclease III-Assisted Signal Amplification Approach.

The detection of aflatoxin B1 (AFB1) has recently garnered much attention on the issue of food safety. In this study, a novel and sensitive aptasensor towards AFB1 is proposed using an Exonuclease III (Exo III)-integrated signal amplification strategy. This reported sensing strategy is regulated by aptamer-functionalized nanobeads that can target AFB1; furthermore, complementary DNA (cDNA) strands can lock the immobilized aptamer strands, preventing the signal amplification function of Exo III in the absence of AFB1. The presence of AFB1 triggers the displacement of cDNA, which will then activate the Exo III-integrated signal amplification procedure, resulting in the generation of a guanine (G)-rich sequence to form a G-4/hemin DNAzyme, which can catalyze the substrate of ABTS to produce a green color. Using this method, a practical detection limit of 0.0032 ng/mL and a dynamic range of detection from 0.0032 to 50 ng/mL were obtained. Additionally, the practical application of the established sensing method for AFB1 in complex matrices was demonstrated through recovery experiments. The recovery rate and relative standard deviations (RSD) in three kinds of cereal samples ranged from 93.83% to 111.58%, and 0.82% to 7.20%, respectively, which were comparable with or better than previously reported methods.

Label-free ratiometric homogeneous electrochemical aptasensor based on hybridization chain reaction for facile and rapid detection of aflatoxin B1 in cereal crops

Aflatoxin B1 (AFB1) contamination has raised global concerns in agricultural and food industry; thus, sensitive, accurate and rapid AFB1 sensors are essential in many circumstances. Herein, we developed a label-free and immobilization-free ratiometric homogeneous electrochemical aptasensor based on hybridization chain reaction (HCR) for facile and rapid determination of AFB1. Methylene blue (MB) and ferrocene (Fc) were used as label-free probes to produce a response signal (IMB) and a reference signal (IFc) in solution phase, respectively. The ratio of IMB/IFc was used as a yardstick to quantify AFB1. HCR was exploited to enlarge the intensity of IMB as well as ratiometric signal. By combining label-free homogeneous assay and ratiometric strategy, the resulting aptasensor offered sensitive, rapid, and reliable determinations of AFB1 with a detection limit of 38.8 pg mL−1. The aptasensor was then used to determine AFB1 in cereal samples with comparable reliability as HPLC-MS.

Application of deep eutectic solvent-based extraction coupled with an S-CQD fluorescent sensor for the determination of pirimicarb in cereals

A novel deep eutectic solvent-based extraction and sulfur-doped carbon quantum dots (S-CQDs) serving as fluorescence probes to detect pirimicarb in cereals were established. The deep eutectic solvent was synthesized using choline chloride and butanediol, achieving direct and efficient extraction of pirimicarb residue in the cereals. The fluorescence quenching of S-CQDs was caused by the electrostatic interaction between the negatively charged S-CQDs and positively charged thiocholine, which was the hydrolysate of acetylthiocholine. The fluorescence of S-CQDs was enhanced as the activity of acetylcholinesterase (AChE) was inhibited by pirimicarb, achieving the detection of pirimicarb in the cereal samples. The limit of detection (LOD) was 0.006 μg mL−1. The recovery ranged from 96.6% to 108.2%. This extraction and detection method of pirimicarb based on an environmentally friendly DES and S-CQD fluorescent sensor maintains good stability and convenience, offering a promising strategy for extracting and testing harmful substances in food samples.

Evaluation of the Methyl and Inorganic Mercury in Infant Formula Milk and Cereals Samples: Estimated Risk Assessment in Children under 2.0 Years

Evaluation of the Methyl and Inorganic Mercury in Infant Formula Milk and Cereals Samples: Estimated Risk Assessment in Children under 2.0 Years

QuEChERS based analysis of multiple pesticides and phthalates in packaged food products

Pesticides and phthalates are toxic chemical compounds which could be present in our food items through their different usage. Some of these toxic analytes are present in traces and always a challenge to quantify for analytical chemist especially from complex food matrices. A simple, sensitive, fast and robust QuEChERS based method was developed, optimized and validated. The method showed good linearity in the range of 0.30–50.00 ng g−1 with (r2) ≥ 0.9855. The quantification limit was in the range of 0.0983–0.8934 ng g−1 with recovery and precision in the range of 71.1–112.5% and RSD < 7.9% for all the analytes. The combined and expanded uncertainty was also determined showing strong analytical significance of the method with negligible matrix interferences. The developed method was successfully used for the analysis of 12 packaged cereal samples (flavoured = 7 & unflavoured = 5). Partial least square discrimination analysis (PLS–DA) of Multivariate approach was used to find differences in profile of pesticides and phthalates between the flavoured and unflavoured cereal samples. We obtained three pesticides (S-BioAllethrin, Dieldrin, Chlorfenapyr) as most important discriminatory compounds for the two groups after comparing fold change (FC) and variable importance in projection (VIP) analysis. Univariate analysis of 8 pesticides obtained from FC analysis showed increased concentration in flavoured group than the unflavoured cereals. The proposed work will be of immense use for routine analysis of these toxic compounds to deal with food fraud and uncover associated risk to the consumer.

A Novel Multiplex Mycotoxin Surface-Enhanced Raman Spectroscopy Immunoassay Using Functional Gold Nanotags on a Silica Photonic Crystal Microsphere Biochip.

A novel multiplex mycotoxin surface-enhanced Raman spectroscopy (SERS) immunoassay was established for the first time on different artificial antigen-modified silica photonic crystal microspheres (SPCMs), which can be integrated into a biochip array to achieve multiplex detection using corresponding antibody-functionalized gold nanoparticles (AuNPs) as the SERS nanotag. The unique optical structure of SPCMs is helpful to find the detection spots easily, accommodate a large amount of probe molecules, and enhance the Raman signal intensity. Such enhancement was confirmed by the simulation result, showing the electric field enhancing effect in SPCMs with AuNPs being 7 times. A competitive SERS immunoassay was established using antigen-modified SPCMs and mycotoxins to compete for binding antibody-functionalized SERS nanotags, displaying broad linear detection ranges of 0.001-0.1 ng/mL for aflatoxin B1 (AFB1), 0.01-10 ng/mL for ochratoxin A (OTA), and 0.001-0.1 ng/mL for zearalenone (ZEN) and low detection limits of 0.82 pg/mL for AFB1, 1.43 pg/mL for OTA, and 1.00 pg/mL for ZEN. In the spiked cereal samples, recovery rates of the method were measured in the range of 70.35-118.04% for the three mycotoxins, which was in agreement with that of the traditional enzyme-linked immunosorbent assay method. The SERS immunoassay for mycotoxin detection also showed high specificity and good repeatability and reproducibility. The new microsphere-based SERS immunoassay biochip only requires a one-step reaction and overcomes the disadvantages of fluorescence and chemiluminescence background signals. The work paves the way for further developing SERS-based microsphere suspension arrays for new targets.

In vitro digestion characteristics of cereal protein concentrates as assessed using a pepsin-pancreatin digestion model

An alkaline extraction method has been used in many studies to extract total protein from cereal samples. Wheat bran protein concentrate (WBPC), oat bran protein concentrate (OBPC), and barley protein concentrate (BPC) were prepared by alkaline extraction and isoelectric precipitation to study their functional and nutritional properties. The three protein concentrates were hydrolysed by an in vitro pepsin-pancreatin digestion model. Their digestibility (%) and degree of hydrolysis (DH%) were evaluated, and SDS-PAGE electrophoresis was used to illustrate the protein and peptides patterns. The change of the particle sizes and the release of the essential amino acids was followed during the digestion process. The in vitro digestibility of WBPC, OBPC and BPC was 87.4%, 96.1% and 76.9%, respectively. The DH% of protein concentrates were between 50 and 60%. The change of the particle size distribution values Dv(50) was assumed to be related to protein aggregations during the digestion. The protein fractions were identified and the degradation during the digestion and were analysed by SDS-PAGE; the gels of WBPC and OBPC digestion showed virtually complete degradation whereas the intensive bands of undigested protein were presented for BPC. The generation of the free amino acids and short chain peptides were significantly higher at the end of the intestinal digestion compared to the stages of before and after gastric digestion. Higher content of the deficient amino acids such as lysine and threonine were found comparing to the level of deficient amino acids in cereal grains but does not meet the daily recommended intake.

Dietary Risk Assessment and Consumer Awareness of Mycotoxins among Household Consumers of Cereals, Nuts and Legumes in North-Central Nigeria.

This study characterized the health risks due to the consumption of mycotoxin-contaminated foods and assessed the consumer awareness level of mycotoxins in households in two north-central Nigerian states during the harvest and storage seasons of 2018. Twenty-six mycotoxins and 121 other microbial and plant metabolites were quantified by LC-MS/MS in 250 samples of cereals, nuts and legumes. Aflatoxins were detected in all food types (cowpea, maize, peanut and sorghum) except in millet. Aflatoxin B1 was the most prevalent mycotoxin in peanut (64%) and rice (57%), while fumonisin B1 occurred most in maize (93%) and beauvericin in sorghum (71%). The total aflatoxin concentration was highest in peanut (max: 8422 µg/kg; mean: 1281 µg/kg) and rice (max: 955 µg/kg; mean: 94 µg/kg), whereas the totals of the B-type fumonisins and citrinin were highest in maize (max: 68,204 µg/kg; mean: 2988 µg/kg) and sorghum (max: 1335 µg/kg; mean: 186 µg/kg), respectively. Citrinin levels also reached 51,195 µg/kg (mean: 2343 µg/kg) in maize. Aflatoxin and citrinin concentrations in maize were significantly (p < 0.05) higher during storage than at harvest. The estimated chronic exposures to aflatoxins, citrinin and fumonisins were high, resulting in as much as 247 new liver cancer cases/year/100,000 population and risks of nephrotoxicity and esophageal cancer, respectively. Children who consumed the foods were the most vulnerable. Mycotoxin co-occurrence was evident, which could increase the health risk of the outcomes. Awareness of mycotoxin issues was generally low among the households.

Prussian blue immunochromatography with portable smartphone-based detection device for zearalenone in cereals

In this study, we developed a prussian blue nanoparticles (PBNPs) immunochromatographic assay (ICA) integrated with smartphone-based detection device for ZEN in cereals. PBNPs, as probe labels, were synthesized with properties of controllable structure, environment friendliness, and high affinities to antibody (Ab). PBNPs-ICA quantitative analysis was performed with a hand-held smartphone-based device coupled with a user-friendly and self-programmed detection App. This integrated strategy demonstrated high sensitivity for ZEN with a cut-off value of 10 μg/kg, a detection limit of 0.12 μg/kg, a quantitation limit of 0.27 μg/kg, and recovery rates of 92.0%-105.0% and 88.0%-98.0% for maize and wheat, respectively. The results of 20 naturally contaminated cereal samples showed good correlation (R2＞0.99) between LC-MS/MS and developed system. Moreover, the stability experiment revealed that PBNPs-ICA maintained high stability and bioactivity against competitive antigen (Ag). The proposed strategy exhibited great potential for the rapid monitoring of mycotoxins or other small molecule hazards.

Development and Validation of an LC-MS/MS Based Method for the Determination of Deoxynivalenol and Its Modified Forms in Maize.

The Fusarium mycotoxin deoxynivalenol (DON) is a common contaminant of cereals and is often co-occurring with its modified forms DON-3-glucoside (D3G), 3-acetyl-DON (3ADON) or 15-acetyl-DON (15ADON). A stable-isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) based method for their determination in cereals was developed and validated for maize. Therefore, 13C-labelled D3G was enzymatically produced using 13C-DON and [13C6Glc]-sucrose and used as an internal standard (IS) for D3G, while uniformly 13C labelled IS was used for the other mycotoxins. Baseline separation was achieved for the critical peak pair DON/D3G, while 3ADON/15ADON could not be fully baseline separated after testing various reversed phase, fluorinated phase and chiral LC columns. After grinding, weighing and extracting the cereal samples, the raw extract was centrifuged and a mixture of the four 13C-labelled ISs was added directly in a microinsert vial. The subsequent analytical run took 7 min, followed by negative electrospray ionization and selected reaction monitoring on a triple quadrupole MS. Maize was used as a complex cereal model matrix for validation. The use of the IS corrected the occurring matrix effects efficiently from 76 to 98% for D3G, from 86 to 103% for DON, from 68 to 100% for 15ADON and from 63 to 96% for 3ADON.

Pesticides Contamination of Cereals and Legumes: Monitoring of Samples Marketed in Italy as a Contribution to Risk Assessment

The evaluation of cereal-based product contamination by pesticide residues is a topic of worldwide importance, and reliable analytical methods for official check analyses and monitoring studies are required for multi-residue analysis at trace levels. In this work, a validated multi-residual analytical method by gas-chromatography and tandem mass spectrometry coupled with a rapid QuEChERS procedure was used for the determination of 37 pesticides (pyrethroids, organophosphorus and organochlorine compounds) in 209 commercially available samples of cereals and 11 legumes, placed on the Italian market in 2018 and 2019, coming from different regions of Italy, eastern Europe, and some non-European countries. No pesticide traces were observed in the analyzed legume samples. A total of 18 cereal samples were found to be contaminated by at least one pesticide, with a concentration level higher than the corresponding quantification limit, but never exceeding the maximum level fixed in the European Regulations. This work is the first part of a surveillance study for pesticide control in food samples.

Limitations of currently available in vitro oestrogenicity bioassays for effect-based testing of whole foods as the basis for decision making

ABSTRACT The idea that previously unknown hazards can be readily revealed in complex mixtures such as foods is a seductive one, giving rise to the hope that data from effect-based assays of food products collected in market surveys is of suitable quality to be the basis for data-driven decision-making. To study this, we undertook a comparative study of the oestrogenicity of blinded cereal samples, both in a number of external testing laboratories and in our own facility. The results clearly showed little variance in the activities of 9 samples when using a single method, but great differences between the activities from each method. Further exploration of these findings suggest that the oestrogenic activity is likely an inherent part of the natural food matrix which the varying sample preparation methods are able to release and extract to differing degrees. These issues indicate the current poor suitability of these types of datasets to be used as the basis for consumer advice or food decision-making. Data quality must be improved before such testing is used in practice.

The use of protein binders and sorghum crisps as potential ingredients in a cereal bar for dogs

AbstractThis study aimed to evaluate the inclusion of different protein binders and sorghum crisps in cereal bars for dogs and their effect on sensory properties, product texture, and dog preference. Fifteen cereal bars were developed in which three crisp sources (rice crisp, white and red sorghum crisp) and five sources of binders (corn syrup, spray dried plasma, gelatin, albumin, and egg product) were evaluated. An interaction effect between binder and crisp sources was found for textural properties (p &lt;.05). A total of 103 volatile compounds were identified and semi‐quantified in the cereal bar samples, with aldehydes being the most represented. Unlike crisp source, protein binders played a major role on sensory properties and impacted the dog's preference. This study suggests that sorghum crisps and protein binders may be used in cereal bars for dogs; however, considerations regarding sensory attributes and dog's preference should be taken to maximize product acceptance.Practical applicationsThis is the first study to report information regarding the use of novel ingredients in a cereal bar application for dogs. The findings observed wherein provide a comprehensive understanding about product development and the impact of ingredients on final product quality, sensory properties, and animal preference. The methodologies and outcomes of our work can be directly translated to the pet food industry to aid in the development of dog treats.