Formation of guaiacol in chocolate milk by the psychrotrophic bacterium Rahnella aquatilis

AIMS: The aim of this study was to identify the causative agent of a smoky/phenolic taint in refrigerated full cream chocolate milk. METHODS AND RESULTS: Microbiological examination of spoiled and unspoiled milk samples from the same processor showed high numbers of the psychrotrophic coliform Rahnella aquatilis in the spoiled samples only. Gas chromatography/mass spectrometry (GC/MS) was used to identify and quantify the taint compound as guaiacol (2-methoxyphenol) in the spoiled milk. Challenge studies in UHT chocolate and white milks inoculated with the isolate and incubated at 4-5 degrees C and 8-9 degrees C for 6 d showed the production of guaiacol in chocolate milk only, which was confirmed and quantified by GC/MS. CONCLUSIONS: The results indicate that if present in refrigerated chocolate milk, Rah. aquatilis can produce guaiacol within the expected shelf-life of the product, even without temperature abuse. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report that the coliform Rah. aquatilis can produce guaiacol in refrigerated chocolate milk products.     

The effects of low fat chocolate milk on postexercise recovery in collegiate athletes

Spaccarotella, KJ and Andzel, WD. The effects of low fat chocolate milk on postexercise recovery in collegiate athletes. J Strength Cond Res 25(12): 3456-3460, 2011-Drinking chocolate milk between exercise sessions may improve recovery. The purpose of this study was to examine the effects of low fat chocolate milk vs. a carbohydrate-electrolyte beverage (CE) on recovery between preseason practice sessions among 5 male and 8 female Division III soccer players. The study used a randomized crossover design: between morning and afternoon practices, athletes received either an amount of chocolate milk that provided 1 g carbohydrate per kilogram body weight or an equal volume of CE (mean volume of 615 ± 101 ml). After their afternoon practice, they completed a shuttle run to fatigue. Data were analyzed using the Wilcoxon paired rank-sign test (for shuttle run time) and the paired samples t-test (for dietary intake). No significant differences in run time were reported for the group. For the men only, there was a trend of increased time to fatigue with chocolate milk compared with the CE (exact p = 0.03). Low fat chocolate milk may therefore be as good as a CE at promoting recovery between training sessions during preseason.     

Chocolate Milk Consequences: A Pilot Study Evaluating the Consequences of Banning Chocolate Milk in School Cafeterias

Objectives

Currently, 68.3% of the milk available in schools is flavored, with chocolate being the most popular (61.6% of all milk). If chocolate milk is removed from a school cafeteria, what will happen to overall milk selection and consumption?

Methods

In a before-after study in 11 Oregon elementary schools, flavored milk–which will be referred to as chocolate milk–was banned from the cafeteria. Milk sales, school enrollment, and data for daily participation in the National School Lunch Program (NSLP) were compared year to date.

Results

Total daily milk sales declined by 9.9% (p<0.01). Although white milk increased by 161.2 cartons per day (p<0.001), 29.4% of this milk was thrown away. Eliminating chocolate milk was also associated with 6.8% fewer students eating school lunches, and although other factors were also involved, this is consistent with the notion of psychological reactance.

Conclusions

Removing chocolate milk from school cafeterias may reduce calorie and sugar consumption, but it may also lead students to take less milk overall, drink less (waste more) of the white milk they do take, and no longer purchase school lunch. Food service managers need to carefully weigh the costs and benefits of eliminating chocolate milk and should consider alternative options that make white milk more convenient, attractive, and normal to choose.     

Transformation of Bacillus subtilis in chocolate milk: evidence for low frequency of establishment of cells transformed under non-selective conditions

Transformation of naturally competent Bacillus subtilis with plasmid was carried out in chocolate milk without antibiotics. Transformed cells were enumerated during the entire growth phase in chocolate milk. When DNA was added to aliquots of a batch culture after different times of incubation, transformation events were detected at all different growth stages. When DNA was added to a batch culture together with the inoculum, transformed cells were detected at the onset of exponential growth. However, apparently no or only limited growth of these transformed cells was observed. To clarify, whether the limitation of growth was due to suppression by non-transformed cells, different proportions of B. subtilis cells either carrying or not carrying the plasmid were mixed and inoculated into chocolate milk without antibiotic. Our results indicate that suppression appears to be of minor importance. Instead, plasmid-bearing cells appear to suffer from a prolonged lag-phase. However, the failure to exhibit significant growth of cells which had taken up the plasmid in chocolate milk appears to be due to failure of these cells to establish themselves as permanently transformed under non-selective conditions.     

Evidence for natural transformation of Bacillus subtilis in foodstuffs

The effect of foodstuffs on the natural transformation of Bacillus subtilis was investigated. As examples of complex food matrices milk with various fat contents as well as chocolate milk were used. The frequencies of transformation varied with the fat content and ranged between 3.8×10⁻⁴ and 1.4×10⁻³. Highest frequencies of about 3×10⁻³ were observed in chocolate milk with 1.5% fat. Development of competence was observed in chocolate milk, resulting in maximal transformation frequencies upon incubation for 10–12 h at 37°C.     

Research of a holiday kind: A clinical trial gone awry: the Chocolate Happiness Undergoing More Pleasantness (CHUMP) study

The randomized controlled trial is the “gold standard” for evaluating the benefits and harms of interventions. The Chocolate Happiness Undergoing More Pleasantness (CHUMP) study was designed to compare the effects of dark chocolate, milk chocolate and normal chocolate consumption on happiness. Although the intention-to-treat analysis showed that participants who received either dark or milk chocolate were happier than those who received no additional chocolate, the actual-consumption analysis showed that there were no differences between any of the groups. The reason for this result is that many participants switched groups mid-study because of their personal chocolate preferences. Although the CHUMP study was pleasurable, it demonstrated the difficulties associated with performing a truly blinded clinical trial.The randomized controlled trial is the “gold standard” for testing the beneficial and harmful effects of interventions. There has been a growing concern in the literature about the potential for bias during randomization of participants in clinical trials.^1–5 I designed the Chocolate Happiness Undergoing More Pleasantness (CHUMP) study to compare the effects of dark chocolate, milk chocolate and “normal” chocolate consumption on happiness. The CHUMP study was a double-blinded clinical trial, and it demonstrated the difficulties associated with performing a truly blinded clinical trial.     

Evaluation of three newly developed direct plating media to enumerate Listeria monocytogenes in foods

LiCl-phenylethanol-moxalactam Agar (LPMA), ARS Modified McBride Agar, and Modified Vogel Johnson Agar were compared with previously tested plating media in the enumeration of Listeria monocytogenes from pasteurized whole milk, chocolate ice cream mix, Brie cheese, and raw cabbage. LPMA was most suitable for analyzing Brie cheese and cabbage. Gum base-nalidixic acid-tryptone-soya medium (previously tested) was most suitable for analyzing milk and chocolate ice cream mix.     

Evaluation of three newly developed direct plating media to enumerate Listeria monocytogenes in foods.

LiCl-phenylethanol-moxalactam Agar (LPMA), ARS Modified McBride Agar, and Modified Vogel Johnson Agar were compared with previously tested plating media in the enumeration of Listeria monocytogenes from pasteurized whole milk, chocolate ice cream mix, Brie cheese, and raw cabbage. LPMA was most suitable for analyzing Brie cheese and cabbage. Gum base-nalidixic acid-tryptone-soya medium (previously tested) was most suitable for analyzing milk and chocolate ice cream mix.     

WHEELCHAIR-MODIFIED SENSORY BOOTHS DO NOT INFLUENCE EVALUATION OF CHOCOLATE PUDDING

The objective of this study was to demonstrate that the dimensions of a sensory booth modified to comply with the Americans with Disabilities Act of 1990 (ADA) guidelines have no impact on consumer evaluations of chocolate pudding. One set of booths each were constructed per ASTM and ADA guidelines. Control pudding was prepared according to package directions with 2% fat milk; other treatments were modified with red food coloring, sugar or nonfat milk. Twenty-four panelists evaluated instant chocolate puddings in directional difference and hedonic tests in each type of booth. Booth type and test date had no effect on judgments.     

Effect of Added Moisture on the Heat Resistance of Salmonella anatum in Milk Chocolate

The heat resistance of Salmonella anatum in milk chocolate at a processing temperature (71 C) was greatly decreased by adding 1 to 4% moisture.     

FLAVOR MODIFICATION OF MILK CHOCOLATE BY CONCHING IN A TWIN-SCREW, CO-ROTATING, CONTINUOUS MIXER

Short-time continuous conching of chocolate offers significant economic advantages over traditional long-time batch methods. High-shear continous mixers can affect conching in a matter of minutes as compared to hours for traditional systems. Milk chocolate conched in two twin-screw, co-rotating, continous mixers operating in series was compared by a variety of sensory methods to chocolate conched by a batch method. A significant difference (P < 0.05) in flavor was found between chocolates conched at 60C by the continuous and batch methods, yet there was no preference for either chocolate. Chocolate conched by the batch method (23 h at 60C) had stronger caramel flavor (P < .10) than chocolate conched continuously, but there were no significant differences (P < .10) in sweet, chocolate or milk flavor. In the continuous system, caramel flavor generally increased with conching temperature and residence time; although, at the highest temperature (95C) and the longest residence time (7.5 min) caramel flavor decreased. Increasing the temperature of continuous conching from 70 to 90C produced chocolate significantly (P < .10) more like chocolate conched in a batch system for 21.5 h at 60C.     

Analysis of cocoa products for ochratoxin A and aflatoxins

Eighty-five samples of cocoa products sampled in Canada were analysed for ochratoxin A (OTA) and aflatoxins in 2011–2012. Inclusion of the aflatoxins in this survey required additional method development. Chocolate was extracted with methanol–water plus NaCl, while for cocoa two successive extractions with methanol and methanol–water were made. Extracts were cleaned on an AflaOchra immunoaffinity column (IAC). Determination was by reversed phase high performance liquid chromatography (HPLC). Detection of the aflatoxins was with a post-column photochemical reactor and of OTA by fluorescence detection. Mean limits of quantification (LOQ) of chocolate and cocoa powders were 0.16 ng/g (OTA) and 0.07 ng/g (aflatoxin B₁), respectively. Survey results showed that the incidences of OTA above the LOQ in natural cocoa were 15/15 (mean 1.17 ng/g), 20/21 for alkalized cocoa (mean 1.06 ng/g), 9/9 for baking chocolate (mean 0.49 ng/g), 20/20 for dark chocolate (mean 0.39 ng/g), 7/10 for milk chocolate (mean 0.19 ng/g), 5/5 for cocoa liquor (mean 0.43 ng/g), and 0/5 for cocoa butter. These results confirm our previous work with OTA. In the same samples, incidences of aflatoxin B₁ above the LOQ were 14/15 for natural cocoa (mean 0.86 ng/g), 20/21 for alkalized cocoa (mean 0.37 ng/g), 7/9 for baking chocolate (mean 0.22 ng/g), 16/20 for dark chocolate (mean 0.19 ng/g), 7/10 for milk chocolate (mean 0.09 ng/g), 4/5 for cocoa liquor (mean 0.43 ng/g), and 0/5 for cocoa butter. Both aflatoxins and OTA were confirmed by HPLC-MS/MS when OTA or aflatoxin levels found were above 2 ng/g in cocoa.     

Selenium content of milk and milk products of Turkey. II

Selenium content of 1028 milk and milk products of Turkey are presented in this study. The selenium content of human milk (colostrum, transitional, and mature milk), various kinds of milk [cow, sheep, goat, buffalo, paper boxes (3%, 1.5%, 0.012% fat), bottled milk, condensed milk (10% fat), mineral added milk (1.6%), and banana, strawberry, and chocolate milk] and milk products (kefir, yogurt, Ayran, various cheese, coffee cream, ice cream, butter, margarine, milk powder, and fruit yogurt) in Turkey were determined by a spectrofluorometric method. The selenium levels of cow milks collected from 57 cities in Turkey were also determined. Selenium levels in cow milk varied with geographical location in Turkey and were found to be lowest for Van and highest for Aksaray. The results [milk (cow, sheep, goat, buffalo and human) and milks products] were compared with literature data from different countries.     

Two-Phase Slug Flow Heat Exchanger for Microbial Thermal Inactivation Research

A continuous two-phase (air-liquid), slug flow, tubular heat exchanger was developed for microbial thermal inactivation research to give exposure times and temperatures in the range of high-temperature, short-time milk pasteurization as well as heat-treated sample volumes of at least 2 ml. The use of air to compartmentalize the liquid in the capillary tubing prevented the development of laminar flow, which enabled precise identification of the residence time of the fastest flowing particles in the heating, holding, and cooling sections of the instrument. Residence time distributions were quantitated by measuring the degree of spreading of radioactive tracers for water, whole milk, chocolate milk, cream, and ice-cream mix with holding temperatures from 50 to 72 C, holding times from 2 to 60 sec, and heating and cooling times of 1.7 sec each. For a holding time of 60 sec and a fastest particle velocity of 10.2 cm/sec, the velocity ratios of the fastest moving particle to the median particle were 1.05, 1.05, 1.10, and 1.13 for whole milk, chocolate milk, cream, and ice-cream mix, respectively. With shorter holding times, these velocity ratios were even closer to unity. These velocity ratios indicated that the instrument would be an effective tool for thermal inactivation research on microorganisms suspended in homogeneous fluids with a viscosity of 15 centipoises or less at the exposure temperature.     

CONFIDENCE ELLIPSES APPLIED TO THE COMPARISON OF SENSORY PROFILES

Our experiment involved seven panels and six chocolates – five dark chocolates and one milk chocolate. The aim of the study was to compare the sensory profiles of the chocolates. A natural question to ask is “Did the panelists detect any differences among the five dark chocolates or did they systematically contrast them with the milk chocolate?” The scatter plot of the chocolates obtained by principal component analysis was useless to answer that question, because of the proximity of the points. To overcome that, we used confidence ellipses calculated using bootstrap. The originality of the study lies in the fact that we applied those ellipses to hierarchical multiple factor analysis (HMFA): among the seven panels, six were composed of trained professionals and the last one was composed of untrained students, and through that method, we managed to compare the two types of panels and balance the role of each trained panel. HMFA provides in a single scatter plot a representation of the six chocolates for each panel, the trained panels and all the panels. Confidence ellipses around each chocolate show that the combined panels – the six trained panels and also the untrained panel – differentiate the five dark chocolates. They also show how much larger the untrained panel's variability is than that of the trained panels, and how comparable are the trained panels' variability to each other.     

Studies on the Thermal Destruction of Escherichia coli in Milk and Milk Products

Escherichia coli (ATCC no. 9637) was shown to have mean D values (slope of destruction rate curve) of 28.2, 5.1, 1.3, 0.00195, 0.00100, 0.00055, 0.00029, and 0.00016 min in milk at temperatures of 125, 130, 135, 168, 170, 172, 174, and 176 F, respectively. The mean D values for this organism in chocolate milk were 32.2, 10.4, 2.6, 0.00265, 0.00133, 0.00069, 0.00035, and 0.00028 min at respective temperatures of 125, 130, 135, 168, 170, 172, 174, and 176 F. Mean D values of 34.4, 10.0, 3.5, 0.00093, 0.00080, 0.00068, 0.00043, and 0.00036 min were found in cream for respective temperatures of 125, 130, 135, 168, 170, 172, 174, and 176 F. In ice cream mix the organism was found to have mean D values of 39.3, 15.2, 5.1, 0.00147, 0.00120, 0.00078, 0.00070, and 0.00053 min for temperatures of 125, 130, 135, 170, 172, 174, 176, and 178 F, respectively. The slopes of the thermal death time curves were found to be 10.2, 10.2, 10.0, and 10.3 F for this organism in milk, chocolate milk, 40% fat cream, and ice cream mix, respectively.     

Thermal Resistance of Certain Oncogenic Viruses Suspended in Milk and Milk Products

Thermal destruction rate curves were determined for adenovirus 12, reovirus 1, and herpes simplex virus in sterile milk, raw milk, raw chocolate milk, and raw ice cream mix. At 40 to 60 C, the curves were asymptotic to the base line. At 65 C, which is near the pasteurization standard, the curves approached a first-order reaction. Thermal resistance studies, by means of in vivo assays, of Moloney and Rauscher leukemia viruses and Moloney and Rous sarcoma viruses indicated that Rous sarcoma was the most resistant. A comparison of the 12D processes of Rous sarcoma virus, reovirus 1, adenovirus 12, and herpes simplex virus in ice cream mix (the most protective of the suspending menstrua studied) with the U.S. Public Health Service pasteurization standard indicated an adequate safety factor in current pasteurization practices.     

Chrysosporium species, potential spoilage organisms of chocolate

Standard methods of analysing foods for the presence of moulds are inadequate for thedetection of genera such as Chrysosporium which do not grow at the high wateractivities of most mycological media. The use of malt, yeast, 50% glucose agar (MY50G) insealed containers as an enrichment medium allowed time for germination and growth ofheat-stressed spores. Three Chrysosporium spp., C. xerophilum Pitt, C. inops (Carmichael) and C. farinicola (Burnside) Skou, were isolated fromcommercial chocolate bars with a water activity (a _w ) of approximately0·28. Chrysosporium inops was isolated from commercial milk crumb and anew Chrysosporium sp. was isolated from Ghanaian cocoa beans. In chocolates madeby coating MY50G agar (a_w = 0·89) with chocolate (a_w = 0·27) containing C. inops arthroconidia, two types of deterioration were seenafter storage. The first was fat bloom due to recrystallization of the cocoa butter on the outer andinner chocolate surface. The second was growth of C. inops which occurred on theinside chocolate surface adjacent to the MY50G agar filling and on the outside surface afterholding at 92% equilibrium relative humidity (erh) for 12 d. There was some evidence that C. inops could grow on the outside of chocolates held at 5·7% erh after 4months' storage at 25 °C. The appearance of the white fungal growth was not unlikefat bloom to the naked eye but was clearly different with the electron microscope.     

Comparison of bacterial counts obtained from naturally contaminated foods by means of Stomacher and blender

Four Regional Health Protection Branch laboratories each compared aerobic colony counts obtained after "stomaching" and blending, for a minimum of 10 samples in each of the seven food groups: dry pastas; chocolate and cocoa powders; frozen entrees (macaroni and cheese, chow mein, chop suey, fried rice, seafood casseroles, and Salisbury steak); nonfat dry milk; shrimp and crabmeats; spices; and breakfast sausages. Overall, counts obtained after using the Stomacher were equivalent to or higher than counts obtained after using the blender in 73% of the comparisons (alpha = 0.05). Where differences existed, counts obtained after using the Stomacher tended to be higher than counts obtained after using the blender from milk powder and lower from sausage. Aerobic colony counts from these foods are not unacceptably biased when obtained by Stomacher.     

NUTRITIONAL, PHYSICAL AND SENSORY CHARACTERISTICS OF VARIOUS CHOCOLATE-FLAVORED PEANUT–SOY BEVERAGE FORMULATIONS

Mixture design and pilot‐plant scale processing protocol were developed to obtain the best chocolate‐flavored peanut–soy beverage formulation. Twenty‐eight formulations were evaluated for nutritional (lysine content), physical (viscosity [η], visual stability index [VSI]) and nine sensory (consumer liking) attributes. Lysine contents (mg/g protein) (44.1–57.1) were close to the reference (51.0) and in the desirable range observed for other peanut‐based beverages. Higher viscosity indicated lower consumer liking, but lower viscosity resulted in lower VSI. Formulation ?8 having 43.9% peanut, 36.3% soy protein isolate (SPI) and 19.8% chocolate syrup had the highest consumer liking and the best balance of physical properties (η = 41.5 mPa·s; VSI = 0.99). As compared to commercial chocolate milk (6 = slightly like to 7 = moderately like), it was rated higher for appearance (7.0), color (6.8) and sweetness (6.4). Two SPI formulations (?6 and ?7) and a soy flour formulation (?14) were rated highest for aroma (6.2), color (7.0) and flavor (6.0), respectively.