Evaluating Paenibacillus odorifer for its potential to reduce shelf life in reworked high-temperature, short-time fluid milk products.

Rework is a common practice used in the dairy industry as a strategy to help minimize waste from processing steps or errors that might otherwise render the product unsaleable. Dairy processors may rework their high-temperature, short-time (HTST) fluid milk products up to code date (21 d) at a typical dilution rate of ≤20% rework into ≥80% fresh raw milk. Bacterial spores present in raw milk that can survive pasteurization and grow at refrigeration temperatures are often responsible for milk spoilage. However, the potential impact of growth and thermal resistance of organisms in reworked product has not been investigated. Our objective was to characterize growth, sporulation, and thermal resistance of Paenibacillus odorifer under conditions representative of extreme storage conditions (time and temperature) of reduced fat (2%) and chocolate milk to evaluate whether product containing rework would have a reduced shelf life. Commercial UHT-pasteurized 2% milk and chocolate milk were independently inoculated with 4 strains of P. odorifer at 1 to 2 log cfu/mL and stored at 4°C and 7°C for 30 d. Changes in P. odorifer cell densities were determined by standard serial dilution with spread plating on tryptic soy agar with yeast extract and incubation at 25°C for 48 h. Spore counts were determined following thermal treatment at 80°C for 12 min. Thermal resistance of a cocktail of P. odorifer in milk was determined after treatments at 63°C for 30 min and 72°C for 15 s. Strains of P. odorifer grew rapidly at 7°C and reached a maximum cell density of ~8 log cfu/g in both 2% and chocolate milk within 12 d. All strains grew more slowly at 4°C and had not reached maximum cell density by 21 d. With extreme temperature abuse (25°C, 24 h), P. odorifer will sporulate in milk; however, thermally resistant subpopulations, including spores, did not develop in milk at 4°C until after stationary phase was achieved (>24 d). Vegetative cells of P. odorifer were verified to be sensitive to pasteurization (>7 log reduction); therefore, P. odorifer would not be expected to contribute to reduced shelf life of fluid milk products containing rework, even with extended storage before rework.

Microbial and chemical composition of Cheddar cheese supplemented with prebiotics from pasteurized milk to aging.

Microbial and chemical properties of cheese is crucial in the dairy industry to understand their effects on cheese quality. Microorganisms within this fat, protein, and water matrix are largely responsible for physiochemical characteristics and associated quality. Prebiotics can be used as an energy source for lactic acid bacteria in cheese by altering the microbial community and provide the potential for value-added foods, with a more stable probiotic population. This research focuses on the addition of fructooligosaccharides (FOS) or inulin to the Cheddar cheese-making process to evaluate the effects on microbial and physicochemical composition changes. Laboratory-scale Cheddar cheese produced in 2 replicates was supplemented with 0 (control), 0.5, 1.0, and 2.0% (wt/wt) of FOS or inulin using 18 L of commercially pasteurized milk. A total of 210 samples (15 samples per replicate of each treatment) were collected from cheese-making procedure and aging period. Analysis for each sample were performed for quantitative analysis of chemical and microbial composition. The prevalence of lactic acid bacteria (log cfu/g) in Cheddar cheese supplemented with FOS (6.34 ± 0.11 and 8.99 ± 0.46; ± standard deviation) or inulin (6.02 ± 0.79 and 9.08 ± 1.00) was significantly higher than the control (5.84 ± 0.27 and 8.48 ± 0.06) in whey and curd, respectively. Fructooligosaccharides supplemented cheeses showed similar chemical properties to the control cheese, whereas inulin-supplemented cheeses exhibited a significantly higher moisture content than FOS and the control groups. Streptococcus and Lactococcus were predominant in all cheeses and 2% inulin and 2% FOS-supplemented cheeses possessed significant amounts of nonstarter lactic acid bacteria found to be an unidentified group of Lactobacillaceae, which emerged after 90 d of aging. In conclusion, this study demonstrates that prebiotic supplementation of Cheddar cheese results in differing microbial and chemical characteristics.

Shifts of microbiota during cheese production: impact on production and quality.

The high-throughput DNA sequencing (HTS) method is used to identify microbes in cheese and their potential functional properties. The technique can be applied to the microbiota of the cheese processing environment, raw milk, curd, whey, and starter cultures, and be used to improve the quality, safety, and other physicochemical properties of the final product. The HTS method is also utilized to study the microbiota shift of different types of cheeses during processing, as the composition and functional properties of the microbiome provide unique characteristics to different cheeses. Although there are several reviews that focused on microbiota of various types of cheeses, this review focuses on evaluating the microbiota shift of different types of cheese production and highlights key bacteria in each step of the processing as well as microbiota of various types of cheeses. KEY POINTS: • High-throughput sequencing can be applied to identify microbiota in cheese. • Microbiota in cheese is changed during making process and aging. • Starter culture plays an important role to establish microbiota in cheese.

Assessment of overall microbial community shift during Cheddar cheese production from raw milk to aging.

Cheese is a fermented dairy product that is made from animal milk and is considered to be a healthy food due to its available nutrients and potential probiotic characteristics. Since the microbes in the cheese matrix directly contribute to the quality and physicochemical properties of cheese, it is important to understand the microbial properties of cheese. In this study, Cheddar cheeses produced on three different dates at the Arbuthnot Dairy Center at Oregon State University were collected to determine the microbial community structure. A total of 773,821 sequencing reads and 271 amplicon sequence variants (ASVs) were acquired from 108 samples. Streptococcus and Lactococcus were observed as the most abundant ASVs in the cheese, which were used as the starter lactic acid bacteria (SLAB). Escherichia coli was detected in the raw milk; however, it was not detected after inoculating with SLAB. According to an alpha diversity analysis, SLAB inoculation decreased the microbial richness by inhibiting the growth of other bacteria present in the milk. A beta diversity analysis showed that microbial communities before the addition of SLAB clustered together, as did the samples from cheese making and aging. Non-starter lactic acid bacteria (NSLAB) were detected 15 weeks into aging for the June 6th and June 26th produced cheeses, and 17 weeks into aging for the cheese produced on April 26th. These NSLAB were identified as an unidentified group of Lactobacillaceae. This study characterizes the changes in the Cheddar cheese microbiome over the course of production from raw milk to a 6-month-aged final product. KEY POINTS: • 271 ASVs were acquired from cheese production from raw milk to 6-month aging. • Addition of SLAB changed the microbial diversity during Cheddar cheese making procedure. • NSLAB were detected more than 15 weeks after aging. Graphical Abstract.

Microbial communities of a variety of cheeses and comparison between core and rind region of cheeses.

Understanding the microbial community of cheese is important in the dairy industry, as the microbiota contributes to the safety, quality, and physicochemical and sensory properties of cheese. In this study, the microbial compositions of different cheeses (Cheddar, provolone, and Swiss cheese) and cheese locations (core, rind, and mixed) collected from the Arbuthnot Dairy Center at Oregon State University were analyzed using 16S rRNA gene amplicon sequencing with the Illumina MiSeq platform (Illumina, San Diego, CA). A total of 225 operational taxonomic units were identified from the 4,675,187 sequencing reads generated. Streptococcus was observed to be the most abundant organism in provolone (72 to 85%) and Swiss (60 to 67%), whereas Lactococcus spp. were found to dominate Cheddar cheese (27 to 76%). Species richness varied significantly by cheese. According to alpha diversity analysis, porter-soaked Cheddar cheese exhibited the highest microbial richness, whereas smoked provolone cheese showed the lowest. Rind regions of each cheese changed color through smoking and soaking for the beverage process. In addition, the microbial diversity of the rind region was higher than the core region because smoking and soaking processes directly contacted the rind region of each cheese. The microbial communities of the samples clustered by cheese, indicated that, within a given type of cheese, microbial compositions were very similar. Moreover, 34 operational taxonomic units were identified as biomarkers for different types of cheese through the linear discriminant analysis effect size method. Last, both carbohydrate and AA metabolites comprised more than 40% of the total functional annotated genes from 9 varieties of cheese samples. This study provides insight into the microbial composition of different types of cheese, as well as various locations within a cheese, which is applicable to its safety and sensory quality.

Information Retrieval in Food Science Research II: Accounting for Relevance When Evaluating Database Performance.

The impetus for this study is the limited amount of information on performance characteristics of food science-encompassing online bibliographic databases. Database usage is an important element in modern research because a comprehensive understanding of current knowledge is essential for effective, unbiased hypothesis formulation and testing. Six databases commonly recommended by academic libraries for information retrieval in the food sciences (Academic Search Premier [ASP], Agricultural Online Access [AGRICOLA], CAB Direct, Food Science and Technology Abstracts [FSTA], PubMed, and Web of Science [WoS]) were compared in a case study based on the research topic "in vitro bile acid binding properties of dietary lignin." A complex string of search terms was used for citation retrieval, and predetermined inclusion/exclusion criteria were used to determine the relevancy of retrieved articles. Searching WoS and CAB Direct returned the greatest number of relevant articles, followed closely by FSTA, PubMed, and AGRICOLA. FSTA and AGRICOLA returned the highest ratios of relevant-to-irrelevant articles, followed closely by CAB Direct. None of the databases, when used alone, recovered all of the relevant articles identified in the study; WoS indexed the highest percentage of relevant articles identified (WoS = 10; total = 19). WoS also had the highest number of relevant articles that were unique to any one database. The thoroughness of searching the complete group of databases was tested by comparison of retrieved citations with those found in relevant review articles, revealing the need for testing overall inclusivity. PRACTICAL APPLICATION: Comprehensive online information retrieval is the most efficient means of accessing current knowledge. Awareness of current knowledge is essential for effective, unbiased decision making in private sector and academic/government-based research endeavors. Thus, online database usage is an essential element of modern food science research. This paper provides tangible examples of the performance characteristics of online bibliographic databases commonly recommended for information retrieval in the food sciences. The paper is written so as to aid the reader in making decisions with respect to database usage for the recovery of topic-relevant peer-reviewed articles germane to their area of research.

Camembert-type cheese quality and safety implications in relation to the timing of high-pressure processing during aging.

Bloomy rind cheeses, including Brie, Camembert, and related varieties, are at high risk of contamination by environmental pathogens during manufacture and ripening. This risk is particularly high during ripening due to open-air exposure of the product. Currently, no kill step is applied after manufacture or post ripening to control food safety risks associated with Listeria monocytogenes contamination. Instead, cheesemakers must rely on sanitation and environmental monitoring to reduce this risk. High-pressure processing (HPP) is a nonthermal food-processing technology that can effectively reduce bacterial contaminants with minimal impact on the organoleptic properties of various foods. The objective of this study was to evaluate HPP as a potential intervention to maintain Camembert cheese quality and reduce risk associated with L. monocytogenes. Timing of HPP treatments (3, 11, and 45 d after manufacture) was based on the growth of L. monocytogenes during Camembert cheese ripening. High-pressure processing treatment of fully ripened cheeses (45 d) resulted in destruction of the surface mold, which caused browning and yellowing of the cheese rind. Applying HPP treatment earlier in the ripening process (11 d) resulted in a similar degradation of cheese appearance, which did not improve with continued ripening. Applying HPP treatment shortly after production (3 d; before the surface flora developed) delayed the development of the cheese rind and the textural ripening of the cheese. This early treatment time also resulted in free whey being expelled from the cheese, creating a firmer body. Applying HPP 11 d after manufacture resulted in >5 log reduction of L. monocytogenes at 450 and 550 MPa with holding times of 10 min. Although HPP was effective at reducing L. monocytogenes associated with bloomy rind cheeses, the quality deterioration would be unacceptable to consumers. Cheesemakers must continue to emphasize sanitation and environmental monitoring to reduce the risk of L. monocytogenes in bloomy rind cheeses.

Volatile aroma composition of distillates produced from fermented sweet and acid whey.

Lactose within whey can be fermented and distilled to produce a potable distilled spirit. The aim of this study was to determine if acid and sweet whey types can be fermented and distilled using similar processes and to investigate differences in volatile aroma compounds for the 2 distillates. Fermentation and distillation of the 2 whey types progressed in a similar manner, using Kluyveromyces marxianus for the initial fermentation and a glass still fitted with a Vigreux column for the subsequent distillation. Ethanol content of the wash (fermented whey) varied considerably following each fermentation and ranged from 1.2 and 2.0% (wt/wt) with no clear trend between acid and sweet whey samples. Volatile aroma compounds were extracted using headspace solid-phase microextraction and identified via gas chromatography-mass spectrometry. Acid and sweet whey distillates contained unique volatile aromatic compounds, and significant differences in compound peak areas were observed. These differences may have an effect upon the organoleptic qualities of spirits produced from whey; therefore, whey source may be an important factor when fermenting and distilling whey.

Influence of cheese-making recipes on the composition and characteristics of Camembert-type cheese.

Bloomy rind cheeses, including Camembert and related varieties, can be produced using alternative processes that vary based on milk preacidification, cutting, curd handling, and ripening parameters. Modification of these parameters creates distinct cheeses such as lactic curd, stabilized curd, and hybrids of the two. The objective of this study was to determine the influence of 5 Camembert-type cheese recipes on the composition and characteristics during ripening. Five varieties of Camembert-type cheese were produced: (1) lactic curd, (2) sweet curd, (3) washed curd, (4) solubilized curd, and (5) stabilized curd. Cheeses were aged at 13°C for 10 d, during the mold growth phase, and 7°C from d 11 until 50. Key quality metrics including texture development, pH (center and surface), and color were monitored throughout shelf-life. Compositional evaluation (d 5; fat, protein, moisture, salt, and minerals) grouped cheeses into 3 categories: (1) lactic curd, (2) sweet and washed curd, and (3) solubilized and stabilized curd. The lactic curd and stabilized curd were consistently the most different varieties for composition and quality metrics. Moisture content of Camembert-type varieties ranged from 53.15 to 57.99%, Ca ranged from 0.23 to 0.45%, and P ranged from 0.21 to 0.40%. All varieties followed the expected pH evolution on the rind and in the paste with the pH of the rind reaching 7 by d 10, and paste pH reaching 7 between 35 and 50 d. The displacement of the paste (distance traveled upon cutting) for the lactic curd was the greatest among the 5 varieties, reaching an average of 27 ± 1.9 mm (mean ± standard error) after 50 d of ripening and 60 min of flow time. The stabilized curd on the other hand traveled the shortest distance, reaching an average of 4 ± 0.4 mm at the same time point. Browning, considered a defect in mold-ripened cheeses, was observed in all varieties, but was most substantial for lactic curd (lightness, L*, decreased from 87.19 to 68.58). Based on these quality metrics the shelf-life of these recipes was estimated with the lactic curd having the shortest, and the stabilized curd having the longest. Examining Camembert-type cheese quality metrics for these 5 varieties can assist cheesemakers during recipe formulation and selection of cheese-making practices to achieve optimum product quality.

Information Retrieval in Food Science Research: A Bibliographic Database Analysis.

The aim of the present research was to ascertain the importance of electronic bibliographic database selection and multiple database usage during the information retrieval phase of research in the food sciences. Six commonly recommended databases were compared with respect to overall journal coverage and journal overlap. Databases were also evaluated with respect to coverage of food science-based journals and the extent of article coverage therein. A case study approach, focused on bile acid/dietary fiber interactions, was used to illustrate the ramifications of database selection/usage when dealing with specific research topics. Databases differed with respect to the breadth of disciplines covered, the total number of journals indexed, the number of food science discipline-specific journals indexed, and the number of articles included per indexed journal. All of the databases contained citations that were unique to the given database. The data resulting from the case study provide an example of the extent to which relevant information may be missed if pertinent databases are not mined. In the present case, over half of the articles retrieved on the focus research topic were unique to a single database. The combined data from this study point to the importance of thoughtful database selection and multiple database usage when comprehensively assessing knowledge in the food sciences. PRACTICAL APPLICATION: This paper provides insights into article database usage for food science-relevant information retrieval. Online information retrieval is an efficient way to assess current knowledge in any of the food science disciplines. Acquired knowledge in turn is the underpinning of effective problem solving; whether it be private sector- or academic/government-based research.

Fermentation and distillation of cheese whey: Carbon dioxide-equivalent emissions and water use in the production of whey spirits and white whiskey.

Whey disposal can be both an environmental and economic challenge for artisanal creameries. Lactose in whey can be fermented to produce ethanol and subsequently distilled. The objective of this study was to use a process-based life cycle analysis to compare carbon dioxide-equivalent (CO2e) emissions and water usage associated with the artisanal or craft production of clear, unaged spirits using whey or malted barley as fermentation substrate. Differences in production were assessed based on key process differences: energy used, water used, distillation by-product disposal, and mass of CO2 produced during fermentation. For this study, whey was assumed removed from the artisanal creamery waste stream. Quantifiable differences were evaluated per 750-mL (45% alcohol by volume) functional unit and expressed as mass-equivalent CO2 emissions (kg of CO2e) and mass of water (kg) used. The CO2e emissions and water usage were quantified using published data, thermodynamic calculations, and mass-balance calculations for a hypothetical distillation system. The process-based life cycle analysis estimated that distillation of fermented whey reduced overall CO2e emissions by 8.4 kg per functional unit and required 0.44 kg less water added into the production process compared with production of a similar clear, unaged spirit using malted barley as substrate. Our preliminary analysis suggests that conversion to distilled whey spirit is a more environmentally responsible approach compared with landfill disposal of whey.

Short communication: Snapshot of industry milk hauling practices in the western United States.

The Pasteurized Milk Ordinance (PMO) mandates milk hauling sanitation and operational practices; however, the use of vague language (i.e., "as needed") and gaps in processes lead to variability in industry practices. Our aim was to characterize industry milk hauling practices and identify areas that may be an unexplained source of contamination in the dairy processing continuum, and communicate this information with industry to cultivate best practices. The objectives of this study were to (1) survey industry hauling sanitation and operation practices in the Pacific Northwest region of the United States, and (2) quantify microbial populations [aerobic plate count (APC), lactic acid bacteria, coliforms] on the internal surfaces of transfer hoses (tanker and receiving bay) to determine their potential contribution to the microbiological quality of raw milk. Eleven facilities (78% response rate) participated in our survey. All facilities surveyed were compliant with the PMO; however, overall milk reception layout, sanitation practices, and routine maintenance greatly varied between facilities. Farm hose samples (n = 115) had significantly higher microbial loads (APC: mean 4.7 log cfu/100 cm2; median 5.1 log cfu/cm2) than receiving hose samples (n = 57; APC: mean: 2.1 log cfu/100 cm2; median 1.9 log cfu/100 cm2). Microbial populations on transfer hose surfaces did not correlate with time since last cleaning for either tanker or receiving bay hoses. Microbial content of farm hoses is likely to reflect the microbial quality of the previous milk transferred through the hose, making on-farm management practices the primary consideration to maintain low microbiological counts downstream. Upon arrival at the processor, 10% of farm hoses were missing caps. Although this did not correlate with elevated microbiological counts, uncapped farm hoses are exposed to the farm environment, provide opportunity for contamination, and are in violation of the PMO. Through observations made during our studies, manual cleaning procedures appear to be a major weakness in hauling practices and need more attention. Recognizing and communicating variability and areas of weakness allows industry to elevate their hauling sanitation and operational practices to maintain optimum milk microbiological quality.

Effect of leaving milk trucks empty and idle for 6 h between raw milk loads.

The US Pasteurized Milk Ordinance (PMO) allows milk tanker trucks to be used repeatedly for 24 h before mandatory clean-in-place cleaning, but no specifications are given for the length of time a tanker can be empty between loads. We defined a worst-case hauling scenario as a hauling vessel left empty and dirty (idle) for extended periods between loads, especially in warm weather. Initial studies were conducted using 5-gallon milk cans (pilot-scale) as a proof-of-concept and to demonstrate that extended idle time intervals could contribute to compromised raw milk quality. Based on pilot-scale results, a commercial hauling study was conducted through partnership with a Pacific Northwest dairy co-op to verify that extended idle times of 6 h between loads have minimal influence on the microbiological populations and enzyme activity in subsequent loads of milk. Milk cans were used to haul raw milk (load 1), emptied, incubated at 30°C for 3, 6, 10, and 20 h, and refilled with commercially pasteurized whole milk (load 2) to measure cross-contamination. For the commercial-scale study, a single tanker was filled with milk from a farm known to have poorer quality milk (farm A, load 1), emptied, and refilled immediately (0 h) or after a delay (6 h) with milk from a farm known to have superior quality milk (farm B, load 2). In both experiments, milk samples were obtained from each farm's bulk tank and from the milk can or tanker before unloading. Each sample was microbiologically assessed for standard plate count (SPC), lactic acid bacteria (LAB), and coliform counts. Selected isolates were assessed for lipolytic and proteolytic activity using spirit blue agar and skim milk agar, respectively. The pilot-scale experiment effectively demonstrated that extended periods of idle (>3 h) of soiled hauling vessels can significantly affect the microbiological quality of raw milk in subsequent loads; however, extended idle times of 6 h or less would not measurably compromise milk quality in subsequent loads in commercial tankers. Current tanker sanitation practices appear to be sufficient for maintaining raw milk SPC, LAB, and coliform levels, which are important measures of milk quality.

Evidence of terroir in milk sourcing and its influence on Cheddar cheese.

The concept of local food is rapidly gaining importance within the United States. The foundation of local food is terroir, which links a food to its production environment. The purpose of this study was to investigate evidence of terroir in milk sourcing and its influence on Cheddar cheese flavor. Specifically, the study was designed to assess if consumers could differentiate between Cheddar cheeses made with milk from different dairy farms. Milk from 5 locations, including single dairy farms and commingled sites, was collected from around the state of Oregon. Using raw and pasteurized counterparts of the milk, Cheddar cheese was made and aged. At 5 and 9mo into aging, Cheddar cheese consumers were asked to group the samples based on perceived similarity/dissimilarity of cheese flavor. Grouping data were subjected to multidimensional scaling and subsequent cluster analysis. Results at 5mo into aging revealed that cheeses made by milk originating from different farms (80km apart) within the same region were perceived as different, whereas cheeses made with milk from neighboring farms (5km apart) were grouped together, irrespective of heat treatment (i.e., raw vs. pasteurized). Cheeses made with commingled milk from different regions grouped together. At 9mo of aging, in contrast, a clear separation of perceived flavor was present between the pasteurized and raw cheese samples, whereas the effect of milk sourcing was less pronounced. These data suggest that the geographical location of the milk source has an effect on the flavor of Cheddar cheese, but that the practices of milk commingling and heat treatment likely reduce the effect of geographical location, particularly as cheese ages.

Short communication: Microbial quality of raw milk following commercial long-distance hauling.

Hauling is a critical part of the commercial milk supply chain, yet very few studies have aimed to understand its effect on raw milk quality. This study focused on the effect of extended-duration tanker use during hauling on raw milk quality at a commercial facility. Standard tanker use [cleaned-in-place (CIP) once per 24h] served as a control and an incremental between-load water rinse with sanitizer treatment (RS) was evaluated to mitigate any effect from extended duration hauling. During this study, 1 commercial truck with 2 trailers was monitored for 10d. The truck collected milk at a large dairy farm, transported the milk to a manufacturing facility, and then returned to the same farm for a second load. Each round-trip journey took between 10 and 12h, allowing for 2 loads per 24-h use period. Following the second delivery, the truck was cleaned by CIP treatment starting a new treatment day. Producer samples were collected from the raw milk bulk tank on the farm before loading milk into the tanker. The same milk was sampled directly out of the tanker truck before unloading at the manufacturer. Effect on individual bacteria count, thermophilic spore count, and preliminary incubation count was quantified through common industry tests. Surface sponge swabs were also used to monitor tanker sanitation and the efficacy of cleaning treatments. Results did not identify a negative effect on raw milk quality due to extended duration hauling. Whereas the addition of RS did not provide any measurable quality benefits for the microbial milk quality, swab results demonstrated that the RS treatment was able to reduce surface bacteria in the tanker, although not to the same level as the full CIP treatment. Based on this study, current CIP practices for long distance milk hauling appear to be effective in mitigating any measurable effect on raw milk quality.

A decision-making tool to determine economic feasibility and break-even prices for artisan cheese operations.

Artisan cheese makers lack access to valid economic data to help them evaluate business opportunities and make important business decisions such as determining cheese pricing structure. The objective of this study was to utilize an economic model to evaluate the net present value (NPV), internal rate of return, and payback period for artisan cheese production at different annual production volumes. The model was also used to determine the minimum retail price necessary to ensure positive NPV for 5 different cheese types produced at 4 different production volumes. Milk type, cheese yield, and aging time all affected variable costs. However, aged cheeses required additional investment for aging space (which needs to be larger for longer aging times), as did lower yield cheeses (by requiring larger-volume equipment for pasteurization and milk handling). As the volume of milk required increased, switching from vat pasteurization to high-temperature, short-time pasteurization was necessary for low-yield cheeses before being required for high-yield cheeses, which causes an additional increase in investment costs. Because of these differences, high-moisture, fresh cow milk cheeses can be sold for about half the price of hard, aged goat milk cheeses at the largest production volume or for about two-thirds the price at the lowest production volume examined. For example, for the given model assumptions, at an annual production of 13,608kg of cheese (30,000 lb), a fresh cow milk mozzarella should be sold at a minimum retail price of $27.29/kg ($12.38/lb), whereas a goat milk Gouda needs a minimum retail price of $49.54/kg ($22.47/lb). Artisan cheese makers should carefully evaluate annual production volumes. Although larger production volumes decrease average fixed cost and improve production efficiency, production can reach volumes where it becomes necessary to sell through distributors. Because distributors might pay as little as 35% of retail price, the retail price needs to be higher to compensate. An artisan cheese company that has not achieved the recognition needed to achieve a premium price may not find distribution through distributors profitable.

Effect of commercial hauling practices and tanker cleaning treatments on raw milk microbiological quality.

Consolidation of the US milk industry has led to use of tankers for up to 24 h in between thorough cleanings. As the heavy use of tankers has not been previously studied, the effect of this form of hauling on raw milk quality is unknown. This study focused on the effect of frequent tanker use during hauling on raw milk quality at a commercial facility. Standard tanker use (cleaned-in-place once per 24 h) served as our control and incremental cleaning treatments (water rinse after each load, water rinse after each load with a sanitizer treatment after 12 h, and 12 h of sanitizer treatment) were added to the study to understand if any effect could be mitigated by more frequent cleaning. Producer samples were collected from the farm before loading milk into the tanker as well as sampling the same milk directly out of the tanker truck before unloading at the manufacturer. The study was repeated at 2 different dairy manufacturing facilities, once during the summer and once during the winter. Milk quality was quantified through industry-relevant microbiological tests: individual bacteria count, thermophilic spore count, and preliminary incubation count. Within the study we defined a negative effect on milk quality as a statistically significant difference between the tanker and producer samples in any of the 3 microbial tests conducted between treatments. Results from the study showed no clear effect due to hauling in individual bacteria count, thermophilic spore count, or preliminary incubation counts. There was also no difference in milk quality between the 2 plants, suggesting that neither season nor location affected our results in the standard use variable. As we did not see a negative effect on milk quality in the standard use variable, the addition of cleaning treatments did not appear to provide any clear benefit. Tanker surface swabs and ATP swabs were also used to monitor tanker sanitation and the efficacy of cleaning treatments. Both surface and ATP swabs revealed differences between cleaning efficacy at the 2 facilities. Although the differences in efficacy did not influence tanker milk quality within our study, variability in sanitation may provide a source of contamination that could negatively affect raw milk quality in other areas. Based on this study, current hauling practices appear to be effective in mitigating any measurable effect on raw milk quality; however, further investigation is needed before making industry-wide recommendations.

Start-up and operating costs for artisan cheese companies.

Lack of valid economic data for artisan cheese making is a serious impediment to developing a realistic business plan and obtaining financing. The objective of this study was to determine approximate start-up and operating costs for an artisan cheese company. In addition, values are provided for the required size of processing and aging facilities associated with specific production volumes. Following in-depth interviews with existing artisan cheese makers, an economic model was developed to predict costs based on input variables such as production volume, production frequency, cheese types, milk types and cost, labor expenses, and financing. Estimated values for start-up cost for processing and aging facility ranged from $267,248 to $623,874 for annual production volumes of 3,402 kg (7,500 lb) and 27,216 kg (60,000 lb), respectively. First-year production costs ranged from $65,245 to $620,094 for the above-mentioned production volumes. It is likely that high start-up and operating costs remain a significant entry barrier for artisan cheese entrepreneurs.

The Mycobacterium avium subsp. paratuberculosis MAP3464 gene encodes an oxidoreductase involved in invasion of bovine epithelial cells through the activation of host cell Cdc42.

Mycobacterium avium subsp. paratuberculosis infection of cattle takes place through the intestinal mucosa. To identify M. avium subsp. paratuberculosis genes associated with the invasion of bovine epithelial cells in vitro, we screened a library of transposon mutants. Several mutants of M. avium subsp. paratuberculosis were identified which invaded Madin-Darby bovine kidney (MDBK) epithelial cells less efficiently than wild-type (wt) M. avium subsp. paratuberculosis. The deltaOx mutant had the transposon located in the MAP3464 gene, a putative oxidoreductase gene whose expression is upregulated upon bacterial contact with MDBK cells. Complete restoration of invasion comparable to that for the wt bacterium was achieved by introducing a copy of the complete oxidoreductase operon into the deltaOx mutant. Immunoprecipitation and Western blot analysis indicated that wt M. avium subsp. paratuberculosis activates Cdc42 and RhoA pathways of internalization 15 and 60 min after infection of the host cell, respectively. The deltaOx mutant, however, failed to activate the Cdc42 pathway. To determine whether an M. avium subsp. paratuberculosis protein delivered to the host cell mediates the entry of the wt bacterium by activation of the Cdc42 pathway, affinity precipitation of active Cdc42 from MDBK-infected cells followed by mass spectrometry was carried out. We identified a 17-amino-acid bacterial peptide associated with the Cdc42 of cells infected with wt M. avium subsp. paratuberculosis but not with the deltaOx mutant. The sequence of the peptide matches MAP3985c, a hypothetical protein, possibly functioning as a putative Cdc42 effector. These findings reveal a novel signaling pathway activated during M. avium subsp. paratuberculosis entry that links the product of MAP3464 gene to activation of Cdc42 in the host cell.

The ability of Mycobacterium avium subsp. paratuberculosis to enter bovine epithelial cells is influenced by preexposure to a hyperosmolar environment and intracellular passage in bovine mammary epithelial cells.

Mycobacterium avium subsp. paratuberculosis is the cause of Johne's disease in cattle and other ruminants. M. avium subsp. paratuberculosis infection of the bovine host is not well understood; however, it is assumed that crossing the bovine intestinal mucosa is important in order for M. avium subsp. paratuberculosis to establish infection. To examine the ability of M. avium subsp. paratuberculosis to infect bovine epithelial cells in vitro, Madin-Darby bovine kidney (MDBK) epithelial cells were exposed to M. avium subsp. paratuberculosis. It was observed that bacteria can establish infection and replicate within MDBK cells. M. avium subsp. paratuberculosis also has been reported to infect mammary tissue and milk, and we showed that M. avium subsp. paratuberculosis infects bovine mammary epithelial cells (MAC-T cell line). Using polarized MAC-T cell monolayers, it was also determined that M. avium subsp. paratuberculosis crosses apical and basolateral surfaces with approximately the same degree of efficiency. Because M. avium subsp. paratuberculosis can be delivered to the naïve host by milk, it was investigated whether incubation of M. avium subsp. paratuberculosis with milk has an effect on invasion of MDBK cells. M. avium subsp. paratuberculosis exposed to milk entered epithelial cells with greater efficiency than M. avium subsp. paratuberculosis exposed to broth medium or water (P < 0.01). Growth of M. avium subsp. paratuberculosis within MAC-T cells also resulted in augmented ability to subsequently infect bovine MDBK cells (P < 0.001). Microarray analysis of intracellular M. avium subsp. paratuberculosis RNA indicates the increased transcription of genes which might be associated with an invasive phenotype.