Milk Spoilage Research Articles

Functional dairy products as a source of bioactive peptides and probiotics: current trends and future prospectives.

Milk is an incredibly healthy food world-wide. However, the 'lactase deficient' individuals cannot digest milk's carbohydrate lactose. A large part of the world population is depriving of highly beneficial milk proteins like casein, lactoalbumin, lactoglobulin, etc. due to lactose intolerance. Production of functional foods and bioactive peptides from milk with natural antioxidants and the addition of probiotics could be the best alternative to extend the use of milk functionalities. Among different probiotics, the lactic acid bacteria (LAB) like Lactobacillus delbrueckii sub sp. bulgaricus, Streptococcus thermophilus and some species of Bifidobacteria and their metabolites (paraprobiotics and postbiotics) have been given more preference to add in milk-derived functional foods. These species are generally considered as heat-tolerant, highly proteolytic, and peptidolytic towards milk proteins and they liberate smaller molecules of bioactive peptides during fermentation and other processes that stimulate the enzyme lactase to help people in digestion of milk carbohydrate lactose. Moreover, the incorporation of natural antioxidants in yoghurt and other dairy products prevents the rancidity of milk fat. The level of bioactive peptides produced in milk-derived functional foods can be determined by capillary zone electrophoresis, mass spectrometry, fractionation, and other modern assessment techniques. Commercial production of functional probiotic products with bioactive peptides could significantly contribute to reduce milk spoilage, enhance health benefits as well as the growth of the agro-processing industry.

Acid–base sensor based on sol–gel encapsulation of bromothymol blue in silica: application for milk spoilage detection

In this work, silica capsules containing bromothymol blue (BTB) were synthesized by a modified Stober sol–gel method. These silica capsules were dispersed into a high-density polyethylene (HDPE)/ethylene-vinyl acetate (EVA) polymeric matrix and then applied in milk spoilage detection tests. The effect of the KOH/BTB molar ratio used in the sol–gel synthesis on the structural, textural, and morphological characteristics of the resulting silica capsules was also investigated. Field emission gun scanning electron microscopy images of the silica capsules showed particles with spherical morphology and size ranging from 0.1 to 1 µm. All silica capsules presented a very low porosity and an encapsulated BTB content in the range of 12–37 µmol g−1. The increase of the KOH/BTB molar ratio resulted in an increase in the relative amount of six-fold siloxane rings and a decrease in the BET surface area of the silica capsules. Spoilage kinetics of milk at room and fridge temperature were much different, as evidenced by pH milk measurements. The milk storage temperature and the KOH content in the silica capsules affected the intensity and rate of blends color change. Blends (HDPE/EVA/silica capsules) with lower KOH content in the capsules showed faster kinetics and higher intensities of color change during the milk spoilage at both tested temperatures. These results showed the potential of the HDPE/EVA/silica capsules as a base material to a sensor packaging, which allows the verification of the milk quality by the end-users.

Riboflavin as an internal marker for spoilage and adulteration detection in milk

Milk is a common consumable in daily life due to its nutritional values. Ensuring milk’s integrity and authenticity is a prime task for researchers and food industries by providing solutions to prevent spoilage and adulteration. We present a robust and reliable optical method to ensure milk quality through its constituent riboflavin as an internal biomarker. Riboflavin is a widely present constituent in several food matrices. This research demonstrates the characteristic fluorescence of riboflavin for checking spoilage and urea adulteration in real-time. The proposed method can even detect and quantify high urea adulteration levels up to 80 mM (i.e., eight times permissible standard value) with a LOD value of 9.3 mM. The linearity (0–80 mM) and high R2 value (0.98, 0.93) of riboflavin’s fluorescence in pure and milk solutions, respectively present this strategy closely associated with fate of milk samples in terms of spoilage and adulteration. Thus, this optical method of riboflavin biosensing in real-time is intuitive and conclusive for determining milk quality.

Octahedral Cuprous Oxide Decorated Flexible Reduced Graphene Oxide Paper for Food Sensing Application

AbstractThis work presents a simple method to fabricate an octahedral cuprous oxide (Cu2O) decorated two‐dimensional (2D) flexible rGOP electrode with filtration and electrodeposition strategies. The characteristic of the Cu2O/rGOP electrodes was recorded by SEM, EDX, XPS, XRD, and Raman spectroscopy. The results clearly showed that Cu2O was successfully electrodeposited on the surface of rGOP by controlling the electrodeposition potential without the introduction of any template or surfactant. The electrochemical characterizations of the Cu2O/rGOP exhibited high electrocatalytic activity toward the reduction of H2O2. The linear detection range for the Cu2O/rGOP flexible sensor was 5.0 μM to 5.5 mM, with a limit of detection of 1.27 μΜ. Subsequently, the developed flexible rGOP sensor was extended for H2O2 detection in milk samples for avoiding milk spoilage. Such judicial preparation of rGOP as a sensing device will certainly pave the way for various other sensing applications including environmental and biomedical applications.

Longitudinal Study of the Bulk Tank Milk Microbiota Reveals Major Temporal Shifts in Composition.

Introduction of microbial contaminations in the dairy value chain starts at the farm level and the initial microbial composition may severely affect the production of high-quality dairy products. Therefore, understanding the farm-to-farm variation and longitudinal shifts in the composition of the bulk tank milk microbiota is fundamental to increase the quality and reduce the spoilage and waste of milk and dairy products. In this study, we performed a double experiment to study long- and short-term longitudinal shifts in microbial composition using 16S rRNA gene amplicon sequencing. We analyzed milk from 37 farms, that had also been investigated two years earlier, to understand the stability and overall microbial changes over a longer time span. In addition, we sampled bulk tank milk from five farms every 1–2 weeks for up to 7 months to observe short-term changes in microbial composition. We demonstrated that a persistent and farm-specific microbiota is found in bulk tank milk and that changes in composition within the same farm are mostly driven by bacterial genera associated with mastitis (e.g., Staphylococcus and Streptococcus). On a long-term, we detected that major shift in milk microbiota were not correlated with farm settings, such as milking system, number of cows and quality of the milk but other factors, such as weather and feeding, may have had a greater impact on the main shifts in composition of the bulk tank milk microbiota. Our results provide new information regarding the ecology of raw milk microbiota at the farm level.

Antimicrobial effects of a bioactive glycolipid on spore-forming spoilage bacteria in milk

The growth of psychrotolerant aerobic spore-forming bacteria during refrigerated storage often results in the spoilage of fluid milk, leading to off-flavors and curdling. Because of their low toxicity, biodegradability, selectivity, and antimicrobial activity over a range of conditions, glycolipids are a novel and promising intervention to control undesirable microbes. The objective of this study was to determine the efficacy of a commercial glycolipid product to inhibit spore germination, spore outgrowth, and the growth of vegetative cells of Paenibacillus odorifer, Bacillus weihenstephanensis, and Viridibacillus arenosi, which are the predominant spore-forming spoilage bacteria in milk. For spore germination and outgrowth assays, varying concentrations (25-400 mg/L) of the glycolipid product were added to commercial UHT whole and skim milk inoculated with ∼4 log10 spores/mL of each bacteria and incubated at 30°C for 5 d. Inhibition of spore germination in inoculated UHT whole milk was only observed for V. arenosi, and only when glycolipid was added at 400 mg/L. However, concentrations of 400 and 200 mg/L markedly inhibited the outgrowth of vegetative cells from spores of P. odorifer and B. weihenstephanensis, respectively. No inhibition of spore germination or outgrowth was observed in inoculated UHT skim milk for any strain at the concentrations tested (25 and 50 mg/L). The effect of glycolipid addition on vegetative cell growth in UHT whole and skim milk when inoculated with ∼4 log10 cfu/mL of each bacteria was also determined over 21 d of storage at 7°C. Glycolipid addition at 50 mg/L was bactericidal against P. odorifer and B. weihenstephanensis in inoculated UHT skim milk through 21 d of storage, whereas 100 mg/L was needed for similar control of V. arenosi. Concentrations of 100 and 200 mg/L inhibited the growth of vegetative cells of B. weihenstephanensis and P. odorifer, respectively, in inoculated UHT whole milk, whereas 200 mg/L was also bactericidal to B. weihenstephanensis. Additional studies are necessary to identify effective concentrations for the inhibition of Viridibacillus spp. growth in whole milk beyond 7 d. Findings from this study demonstrate that natural glycolipids have the potential to inhibit the growth of dairy-spoilage bacteria and extend the shelf life of milk.

Colorimetric indicator based on purple tomato anthocyanins and chitosan for application in intelligent packaging

Intelligent colorimetric indicator films were prepared to monitor freshness/spoilage of milk and fish by incorporating purple tomato anthocyanin (PTA) into chitosan (CS) matrix via solution casting method with PTA concentration (w/w, based on CS) of 10%, 30%, and 50%, respectively. The pH-response, UV absorption, Swelling Index, and the mechanical properties of CS/PTA films were determined. It was found that the color of the original CS/PTA films became darker with an improvement of PTA content and expressed well pH-sensitivity. With increasing of pH, the color of the CS/PTA films exposed to pH = 3–11 solutions became darker and the change in color of the CS/10% PTA film was the most discernable. The tensile strength and Young's modulus of the CS/PTA film was much lower than that of CS film, however, the elongation at breaking and Swelling Index were both improved by adding PTA. The intelligent films with 10% PTA changed their color during progressive spoilage of milk or fish, revealing their potential application for monitoring food freshness/spoilage.

Pattern formation of dried droplets of milk during different processes and classifying them using artificial neural networks

Two methods, one experimental and one based on artificial intelligence/deep learning (AI/DL) are explored in this paper for possible identification of adulteration, coagulation and spoilage of dairy milk. The unique patterns formed by dairy-milk colloids under three conditions – dilution, coagulation and staling and observed using confocal laser scanning microscopy are reported here. The patterns act as a fingerprint to the various stages of coagulation/dilution/spoilage of milk colloids. A deep-learning algorithm is then applied to the patterns thus obtained, and the results of AI/DL based identification of patterns is reported here.

Digital analysis of polydiacetylene quality tags for contactless monitoring of milk

The incorporation of colorimetric sensors as quality indicators in food packaging is an exciting new area of research that could improve food management. The standard approach, however, demands a reliable interface between the sensor and the food and risks food contamination which is a significant consumer concern. To overcome this challenge, herein, we develop a polydiacetylene/phospholipid agarose-based sensor that encapsulates milk in the hydrogel matrix during synthesis. The chemical recognition of free fatty acids, a product of microbial spoilage of the encapsulated milk, induces a gradual blue to red color change in the sensor. We demonstrate that the new composite material exhibits the same spoilage kinetics as regular liquid milk (digital colorimetric response 28 ± 1% and 27 ± 3% respectively), indicating the agarose does not preserve the milk ingredients nor inhibit the detection mechanism of the polydiacetylene sensors. As a result, this sensor can be attached to the external surface of food packaging to provide an indirect indication of food quality without the need for contact with the milk product. The quality tags we present can be “switched” on and off using dehydration and rehydration, removing the need for in situ manufacturing and allowing storage before use. We show that the quality tags produce a similar digital colorimetric response of 21 ± 2% to indicate milk spoilage after rehydration. The color change of the quality tags could not be analyzed using absorption spectroscopy, the standard technique for polydiacetylenes, due to the opacity that milk imparts on the sample. To solve this problem, we develop digital colorimetric analysis software using the Python programming language to describe the extent of color change in polydiacetylene materials and develop a new metric termed the Digital Colorimetric Response that describes polydiacetylene response with excellent linearity (R2 = 0.96). The software is programmed to employ statistical cleaning techniques that automatically remove image noise and outliers based on a pixel’s grayscale Z-score. This new approach to sensor design increases practicality and could be extended to the contactless quality monitoring of other foods, medicines and other products whose safety or quality is jeopardized with direct sensor contact.

Milk Spoilage Detection by Impedance Measurement

Milk Spoilage Detection by Impedance Measurement

Corrigendum to: 'First ever report on the antimicrobial activities of some selected mangrove halophytes of Sundarban against milk spoilage microflora' [Braz. J. Biol. Sci. (2017), vol. 4, no. 8, p. 273-292

The first author reports an error in the original version of this article. In the Figure 2 some of the names of the mangroves have been incorrectly assigned to their respective pictures.

Cattle milk processing and marketing characteristics among agro-pastoralists in North West Nigeria

The study was aimed at analysing the fundamental characteristics of milk collection, processing and marketing among agro-pastoralists in North West Nigeria. A three stage (multi-stage) sampling technique was used for the study. The first stage is the purposive selection of four States (Jigawa, Kaduna, Kano and Katsina States purposively selected based on the high concentration of agro-pastoralists). In the second stage, two Local Government Areas (LGA) from each of the State were purposively selected. The third stage involved the use of snowball sampling method for the selection of communities where agropastoralist were interviewed using structured questionnaire. Two communities were selected per LGA. Data were collected from 157 respondents from the target populatuion out of sampled agro-pastoral households. The survey instrument was pretested and subjected to face validity. The data obtained were analysed using descriptive statistics of frequency counts and percentages. The average length of time for milk storage before processing was 1 hour and 41 minutes. Pasteurization was the method of choice used to prevent milk spoilage by 63.8% of respondents in all the States considered in this study. Almost all (91.2%) of the respondents market their milks. Spoilage of milk during the period before processing was reported as the highest constraint to milk production, processing and marketing in all the four States. Inadequate equipment and milk loss during processing were also reported as constraints by 18.9 and 14.7%, respectively. Low quality of milk, especially during dry season, also constitute a constraint to about 4% of the respondents. The agro-pastoralists in North West Nigeria are engaged in peasantry profitable ventures of milk processing and marketing. Most of them are not members of groups and cooperatives. Their volume of trade in milk processing and marketing can be greatly enhanced if they are organized into productive groups along the value chain. Past efforts at organizing milk processors and marketers into groups and cooperatives need to be revisited by relevant agencies of government and non-government organizations.

Phenotypic and Genotypic Investigation of Two Representative Strains of Microbacterium Species Isolated From Micro-Filtered Milk: Growth Capacity and Spoilage-Potential Assessment.

The microbiota that spoil long-life micro-filtered milk generally includes species of the genus Microbacterium. The metabolic properties of this of microorganisms that could potentially modify the quality of micro-filtered milk are still unexplored when compared to better-known microorganisms, such as the spore-forming Bacillus and Paenibacillus spp., and Gram-negative contaminants, such as species of the genera Pseudomonas and Acinetobacter. In this preliminary study, two strains of Microbacterium (M. lacticum 18H and Microbacterium sp. 2C) isolated from micro-filtered milk were characterized in depth, both phenotypically and genotypically, to better understand their role in long-term milk spoilage. The study highlights the ability of these strains to produce high cell numbers and low acidification in micro-filtered milk under storage and shelf-life conditions. Phenotypic analyses of the two Microbacterium sp. isolates revealed that both strains have low proteolytic and lipolytic activity. In addition, they have the ability to form biofilms. This study aims to be a preliminary investigation of milk-adapted strains of the Microbacterium genus, which are able to grow to high cellular levels and perform slight but not negligible acidification that could pose a potential risk to the final quality of micro-filtered milk. Furthermore, M. lacticum 18H and Microbacterium sp. 2C were genotypically characterized in relation to the characteristics of interest in the milk environment. Some protein-encoding genes involved in lactose metabolism were found in the genomes, such as β-galactosidase, lactose permease, and L-lactate dehydrogenase. The phenotypically verified proteolytic ability was supported in the genomes by several genes that encode for proteases, peptidases, and peptide transferases.

Application of pH Indicator Label based on Beetroot Color for Determination of Milk Freshness

Introduction: Applying of a new indicator in food packaging can be effective to inform consumers about the freshness and quality of the products. Material and Methods: In the current study, a new milk freshness label was investigated containing beetroot color and multi layers of polystyrene. The label characteristics were investigated by estimating color number, release test, and scanning electron microscopy (SEM). The total bacterial count, pH, lactic acid concentration in milk, and label color number were measured using standard plate count, pH meter, titration of acidity, color analysis software, and UV spectrophotometry on days 0, 3, 4, 5, 6, and 7 at refrigerator temperature (4 ± 0.2°C). Results: The label reacted to total bacterial count and pH changes with a visible color during milk spoilage. A positive correlation was found between the label color changes, total bacterial count, and pH. The color of label turned from dark red to light brown, which was related to the chemical changes and bacterial count of milk. Conclusion: According to this simple, visible, and affordable label, the shelf life of pasteurized milk was estimated as five days.

Dairy antioxidant system

A review of dairy antioxidants was done based on local and international sources. Oxidative processes of fat are the main cause of chemical spoilage of milk and dairy products. Lipid oxidation is associated mainly with the amount of unsaturated fatty acids in the shells of fat globules and plasma phospholipids, as well as with the amount of free fat. The products of oxidative reactions at different stages of lipid oxidation with atmospheric oxygen are fatty acid hydroperoxides, peroxides, aldehydes, ketones, hydroxy acidswhich not only worsen the organoleptic characteristics of milk, but some of them are toxic substances. The antioxidant ability of milk is due to vitamins A, C, E, carotenoids, enzymes, lactoferin, sulfur-containing amino acids. One of the most powerful natural antioxidants is ascorbic acid, which is able to absorb superoxide anion radicals, alkoxyl radicals, superoxide, iron oxide, nitric oxide. Group E vitaminsare described, being the primary fat-soluble antioxidants. Among this group of substances, the most biologically active form is ?-tocopherol. The contribution of carotenoids to antioxidant protection has been determined. Beta carotene is considered as a preventive antioxidant, as it can block singlet oxygen which ishighly toxic. The mechanisms of antioxidant action of various enzymes are disclosed. Xanthioxidase has been shown to oxidize various aldigides and purine bases to the corresponding acids. Lactoperoxidase catalyzes the oxidation of thiocyanate with hydrogen peroxide to form thiociagen. Catalase oxidizes hydrogen peroxide to form water and molecular oxygen. Glutathione peroxidase is capable of rapidly removing hydrogen peroxide and other peroxides. The mechanism of the antioxidant action of lactoferrin is described which is based on iron the chelation by it. The functions of sulfur-containing amino acids due to the binding of free radicals are disclosed. Determining the total antioxidant activity can be a biomarker of its biological value, which will allow to choose the most perspective areas of milk processing..

Detection of spore forming Paenibacillus macerans in raw milk

Paenibacillus macerans can cause spoilage of milk during extended storage. However, the natural milk microbiota interferes with the enumeration of Paenibacillus species in raw milk. In this study, a qualitative SYBR Green real-time PCR assay based on the groEL gene was developed for detecting P. macerans (PMassay) in raw milk and compared with one designed for total Paenibacillus detection (TPassay). The specificity of the PMassay was confirmed against a panel of dairy-related spore forming isolates. In the presence of background DNA substituted up to 95%, P. macerans DNA could still be detected by the PMassay although interference occurred as non-target DNA substitution increased. The PMassay was sensitive (detection limit of 2 log CFU/ml in milk) and specific as non-P. macerans isolates gave a Ct > 30. After enrichment of raw milk for 7 days at 37 °C in Reinforced Clostridial Medium with D-cycloserine (RCM-D) under anaerobiosis, Paenibacillus was detected in 10 of the 16 raw milk samples tested. Enrichment in RCM-D yielded about 0.5 to 5.8 log CFU/ml total Paenibacillus and 0.3 to 4.6 log CFU/ml P. macerans in the samples. The assay could be useful in commercial settings, allowing a sensitive detection of P. macerans.

Constraints faced by the dairy farmers in production and marketing of milk in northern dry zone of Karnataka

The present study investigated production and marketing related constraints of dairy farming practice in Northern Dry Zone of Karnataka. The study covered 240 sample households selected randomly from four districts of Northern Dry Zone of Karnataka. About 66 per cent of dairy farmers belongs to marginal (32.50%) and small (33.75%) category. The study also revealed that the 93.75 per cent, 91.66 per cent and 87.91 per cent dairy farmers were facing high cost of feed and fodder, non-availability of quality feed round the year and susceptibility of crossbred animals to disease were the major production related problem, respectively. It was also noted that, the distant location of milk collection center (96%), low price of milk (92.50%), inadequate availability of regular market (91.25%), spoilage of milk due to poor hygiene and storage problem while carrying milk to procurement centre (83.33%) were the major constraints in marketing of milk. Thus efforts should be focused on making availability of quality feed round the year along with the improvement of marketing infrasture is win-win situation for development of dairy sector in the Northern Dry Zone of Karnataka.

Anthocyanin-based sensors derived from food waste as an active use-by date indicator for milk

Active use-by date (AUBD) or freshness indicators hold great potential to reduce food waste. Herein, we develop an anthocyanin AUBD indicator that is capable of discriminating between fresh, spoiling, and spoiled milk. The sensor undergoes a visible blue to purple to pink color change in response to lactic acid, which is an indicator of microbial spoilage in milk. Anthocyanin is cast into a range of materials and the composite’s suitability to monitor pH changes (pH 6.8 fresh milk vs pH 4.0 spoiled milk) is assessed. Of the materials studied, an anthocyanin-agarose film is nominated as the optimum materials with the best colorimetric performance. We introduce a new method to quantify anthocyanin color change by measuring red chromatic shift by digital analysis. The anthocyanin sensors will provide a real-time indication of actual milk quality, surpassing the function of traditional date marking tools that provide an indication of the expected shelf life.

Short communication: Dynamic changes in bacterial diversity during the production of powdered infant formula by PCR-DGGE and high-throughput sequencing

Microorganisms such as thermophilic and psychrotrophic bacteria cause spoilage of milk and milk products [e.g., powdered infant formula (PIF)], mainly because they produce heat-stable extracellular enzymes. However, the dynamic changes in microbial diversity during PIF production are still not well understood. We used denaturing gradient gel electrophoresis (DGGE) and high-throughput sequencing (HTS) to investigate bacterial community structure and distribution during the major stages of PIF production: raw milk, pasteurization, mixing, evaporation, and spray-drying. Our PCR-DGGE analysis indicated that Lactobacillus and Pseudomonas spp. were the dominant bacteria at the raw milk and pasteurization stages; Lactococcus, Streptococcus, Enterococcus, and Lactobacillus spp. were abundant during mixing, evaporation, and spray-drying. Our HTS analysis showed that Pseudomonas had an abundance of 96.79% at the raw milk stage. Lactobacillus, Streptococcus, Thermus, Acinetobacter, and Bacteroides spp. were most common after pasteurization. The index of bacterial diversity was highest at the evaporation stage, suggesting a high potential risk of microbial contamination. The results from DGGE and HTS were consistent in reflecting changes in dominant flora, but different in reflecting the richness of bacterial communities present during PIF production: HTS revealed a much higher richness of bacterial species than DGGE. Our findings from DGGE and HTS showed that psychrophilic and thermophilic bacteria were the main flora present during PIF production: psychrophilic bacteria were mainly Pseudomonas spp. and thermophilic bacteria were mainly Lactobacillus, Streptococcus, and Bacillus spp. To our knowledge, this is the first study to report dynamic changes in microbial communities during PIF production. Our results provide insight into bacterial communities and identify potential contamination sources that could serve as a guide for reducing microbial risk.

Milking time hygiene interventions on dairy farms reduce spore counts in raw milk

Spore-forming bacteria, such as Paenibacillus spp. and Bacillus spp., can survive HTST pasteurization in spore form and affect the quality of dairy products (e.g., spoilage in fluid milk). With the demand for higher quality finished products that have longer shelf lives and that can be distributed further and to new markets, dairy processors are becoming interested in obtaining low-spore raw milk supplies. One method to reduce spores in the dairy system will require disrupting the transmission of spores from environmental locations, where they are often found at high concentrations (e.g., manure, bedding), into bulk tank raw milk. Previous research has suggested that cow hygiene factors (e.g., udder hygiene, level of spores in milk from individual cows) are important for the transmission of spores into bulk tank raw milk, suggesting that one potential strategy to reduce spores in bulk tank milk should target cow hygiene in the parlor. To that end, we conducted a study on 5 New York dairy farms over a 15-mo period to evaluate the effect of a combination of intervention strategies, applied together, on the levels of aerobic spores in bulk tank raw milk. The combination of interventions included (1) training milking staff to focus on teat-end cleaning during milking preparation, and (2) implementing changes in laundered towel preparation (i.e., use of detergent, chlorine bleach, and drying). Study design involved collecting bulk tank raw milk samples for a week before and a week after initiating the combination of interventions (i.e., training on the importance of teat-end cleaning and towel treatment). Observations on teat-end condition, udder hygiene scores, and number of kickoffs during milking were also collected for 24 h before and after implementation of the interventions. A total of 355 bulk tank raw milk samples were collected with mean mesophilic and thermophilic spore counts of 2.1 and 2.4 cfu/mL, respectively, before interventions were applied, and 1.6 and 1.5 cfu/mL, respectively, after the interventions were applied. These reductions represent decreases of 37 and 40% in bulk tank raw milk mesophilic spores and thermophilic spores, respectively. Importantly, spore reductions were observed during each of the 3 visits once the interventions were applied, and the largest reduction in spores was recorded for the first sampling after training the milking staff. Further, when a higher proportion of very rough teat ends was observed, bulk tank milk thermophilic spore counts were significantly higher. The intervention strategies tested here represent easy-to-execute cleaning strategies (e.g., focusing on teat-end hygiene and towel washing procedures) that can reduce bulk tank raw milk spore levels. Future studies should validate the effect of on-farm interventions for reduced spore raw milk on corresponding processed product quality and will need to verify the effects of these small changes on product shelf life.