High temperature, short time pasteurization temperatures inversely affect bacterial numbers during refrigerated storage of pasteurized fluid milk

The grade A Pasteurized Milk Ordinance specifies minimum processing conditions of 72°C for at least 15 s for high temperature, short time (HTST) pasteurized milk products. Currently, many US milk-processing plants exceed these minimum requirements for fluid milk products. To test the effect of pasteurization temperatures on bacterial numbers in HTST pasteurized milk, 2% fat raw milk was heated to 60°C, homogenized, and treated for 25 s at 1 of 4 different temperatures (72.9, 77.2, 79.9, or 85.2°C) and then held at 6°C for 21 d. Aerobic plate counts were monitored in pasteurized milk samples at d 1, 7, 14, and 21 postprocessing. Bacterial numbers in milk processed at 72.9°C were lower than in milk processed at 85.2°C on each sampling day, indicating that HTST fluid milk-processing temperatures significantly affected bacterial numbers in fluid milk. To assess the microbial ecology of the different milk samples during refrigerated storage, a total of 490 psychrotolerant endospore-forming bacteria were identified using DNA sequence-based subtyping methods. Regardless of processing temperature, >85% of the isolates characterized at d 0, 1, and 7 postprocessing were of the genus Bacillus, whereas more than 92% of isolates characterized at d 14 and 21 postprocessing were of the genus Paenibacillus, indicating that the predominant genera present in HTST-processed milk shifted from Bacillus spp. to Paenibacillus spp. during refrigerated storage. In summary, 1) HTST processing temperatures affected bacterial numbers in refrigerated milk, with higher bacterial numbers in milk processed at higher temperatures; 2) no significant association was observed between genus isolated and pasteurization temperature, suggesting that the genera were not differentially affected by the different processing temperatures; and 3) although typically present at low numbers in raw milk, Paenibacillus spp. are capable of growing to numbers that can exceed Pasteurized Milk Ordinance limits in pasteurized, refrigerated milk.     

Effect of pasteurization on survival of Mycobacterium paratuberculosis in milk

Mycobacterium paratuberculosis (Mptb) is the causative agent of Johne's disease of ruminant animals including cattle, goats, and sheep. It has been suggested that this organism is associated with Crohn's disease in humans, and milk is a potential source of human exposure to this organism. A total of 18, including 7 regular batch and 11 high temperature short time (HTST) pasteurization experiments, were conducted in this study. Raw milk or ultra-high temperature pasteurized milk samples were spiked at levels of 10(3), 10(5), and 10(7) cfu of Mptb/ml. Escherichia coli and Mycobacterium bovis BCG strains at 10(7) cfu/ml were used as controls. Pasteurization experiments were conducted using time and temperature standards specified in the Canadian National Dairy Code: regular batch pasteurization method: 63 degrees C for 30 min, and HTST method: 72 degrees C for 15 s. The death curve of this organism was assessed at 63 degrees C. No survivors were detected after 15 min. Each spiked sample was cultured in Middlebrook 7H9 culture broth and Middlebrook 7H11 agar slants. Samples selected from 15 experiments were also subjected to BACTEC culture procedure. Survival of Mptb was confirmed by IS900-based PCR of colonies recovered on slants. No survivors were detected from any of the slants or broths corresponding to the seven regular batch pasteurization trials. Mptb survivors were detected in two of the 11 HTST experiments. One was by both slant and broth culture for the sample spiked to 10(7) cfu/ml of Mptb, while the other was detected by BACTEC for the sample spiked to 10(5) cfu/ml. These results indicate that Mptb may survive HTST pasteurization when present at > or = 10(5) cfu/ml in milk. A total of 710 retail milk samples collected from retail store and dairy plants in southwest Ontario were tested by nested IS900 PCR for the presence of Mptb. Fifteen percent of these samples (n = 110) were positive. However, no survivors were isolated from the broth and agar cultures of 44 PCR positive and 200 PCR negative retail milk samples. The lack of recovery of live Mptb from the retail milk samples tested may be due to either the absence of live Mptb in the retail milk samples tested or the presence of low number of viable Mptb which were undetected by the culture method used in this study.     

The survival of foot-and-mouth disease virus in raw and pasteurized milk and milk products

The Foot-and-Mouth Disease virus (FMDV) is not a public health threat, but it is highly contagious to cloven-footed animals. The virus is shed into milk up to 33 h before there are apparent signs of the disease in dairy cows, and, in extreme cases, signs of disease may not appear for up to 14 d. During this time, raw milk can serve as a vector for spread of the disease both at the farm and during transport to the processing plant by milk tanker. Raw milk and milk products fed to animals have the potential to cause infection, but the potential for pasteurized milk products to cause infection is largely unknown. Current minimum pasteurization standards may not be adequate to eliminate FMDV in milk completely. The purpose of this paper is to assess the literature on the thermal resistance of FMDV in milk and milk products, to identify the risks associated with ingestion of pasteurized products by animals, and to lay a strategy to prevent the spread of FMDV from contaminated milk.     

不同巴氏杀菌处理对绵羊乳热加工特性的影响

目的探究绵羊液态乳的热加工特性。方法以牛乳为对照,分别通过低温长时杀菌((65±2)℃/30 min)、高温短时杀菌(85±2)℃/15 s)和超巴氏杀菌((121±2)℃/5 s)对绵羊乳进行杀菌处理,分析3种巴氏杀菌方式对绵羊乳pH值、黏度、蛋白沉淀量、酪蛋白粒径、zeta电位的影响,评价其热稳定性。结果相比于未处理组,3种巴氏杀菌处理均会显著增加绵羊乳pH值,超巴氏杀菌处理会引起绵羊乳黏度及蛋白沉淀量显著增大(P0.05);在高温短时和超巴氏杀菌处理下,绵羊乳酪蛋白粒径显著增加(P0.05),分别达到185.33 nm和242.70 nm。不同巴氏杀菌绵羊乳间Zeta电位无显著性差异;在同样处理条件下,牛乳黏度、蛋白沉淀量及酪蛋白粒径变化程度均小于绵羊乳。结论绵羊乳热加工特性较牛乳脆弱,加工温度易引起品质劣变,低温长时杀菌方式对绵羊乳的理化特性影响最小。     

不同巴氏杀菌处理对绵羊乳总蛋白及乳清蛋白二级结构的影响

目的: 分析不同巴氏杀菌处理下绵羊乳蛋白的结构变化,同时以牛乳为对照,比较二者差异性。 方法 :分别以低温长时巴氏杀菌（65 ℃ 30 min）、高温短时巴氏杀菌（85 ℃ 15 s）和超巴氏杀菌（121 ℃ 5 s）对绵羊乳、牛乳进行杀菌处理,探究巴氏杀菌处理对绵羊乳和牛乳总蛋白及乳清蛋白二级结构的影响。 结果: 3种巴氏杀菌处理均会影响绵羊乳总蛋白和乳清蛋白二级结构,改变乳蛋白原空间结构;相比总蛋白,乳清蛋白二级结构稳定性受热处理影响更大（P＜0.05,超巴氏杀菌处理对绵羊乳总蛋白与乳清蛋白二级结构影响最大,乳清蛋白变性程度最大。绵羊乳与牛乳在3种巴氏杀菌处理方式下蛋白二级结构的变化规律略有差异。 结论: 3种巴氏杀菌处理下低温长时巴氏杀菌绵羊乳蛋白结构及组成变化最小,且绵羊乳总蛋白与乳清蛋白二级结构的变化规律与牛乳略有不同,在绵羊乳液态奶加工时应引起足够重视。     

Evaluation of calf milk pasteurization systems on 6 Pennsylvania dairy farms

Waste milk has been fed to calves for many years, but concerns with bacterial contamination as well as possible transmission of diseases have discouraged widespread use of this feed. Pasteurization of waste milk is one option to reduce management risk while utilizing a valuable, low-cost, liquid feed source for calves. However, many farms currently pasteurizing waste milk lack a system to adequately monitor the efficiency of the process. A study was carried out to evaluate 6 on-farm pasteurization systems, including high-temperature, short-time pasteurizers and low-temperature, batch pasteurizers. Milk samples were taken pre- and postpasteurization as well as from the calf buckets and immediately frozen for later bacterial culture. Samples were collected twice daily for 15 d. Milk samples were examined for standard plate count (SPC), coagulase-negative staphylococci count, environmental streptococci count, coliform count, gram-negative noncoliform count, Streptococcus agalactiae count, and Staphylococcus aureus count. Before pasteurization, 68% of the samples had SPC <20,000 cfu/mL, and 39% of samples contained <100 cfu/mL of coliform bacteria. After pasteurization, 96% of samples had SPC <20,000 cfu/mL, and 92% had coliform counts <100 cfu/mL. Bacteria counts were significantly reduced by pasteurization, and pasteurized milk contained acceptable numbers of bacteria in >90% of samples. These results indicate that pasteurization can be very effective in lowering bacterial contamination of milk. However, bacteria numbers significantly increased after pasteurization and, in some cases, bacteria counts in milk fed to calves were similar to prepasteurization levels. Milk handling after pasteurization was identified as an important issue on the farms studied.     

巴氏杀菌中性乳饮料防腐保鲜研究

研究了巴氏杀菌中性乳饮料的防腐保鲜技术。采用以鱼精蛋白为主要成分,结合其他防腐剂的优点,复配出中性食品复合防腐剂(NFCP),并应用于巴氏杀菌甜牛乳饮料中。结果表明,在经过100℃,30min杀菌的甜牛乳饮料中,加入质量分数为0.9%的中性食品复合防腐剂,能起到较好的防腐保鲜效果,保质期可达到6个月以上。     

Effect of waste milk pasteurization on fecal shedding of Salmonella in preweaned calves

The objective of the current research was to determine if pasteurization of nonsaleable waste milk influences fecal Salmonella concentrations and prevalence, or antimicrobial susceptibility and serotype of the cultured isolates. Holstein dairy calves (n = 211) were housed on a single commercial dairy in the southwestern United States and randomly allotted to be fed either pasteurized (PWM; n = 128 calves) or nonpasteurized waste milk (NPWM; n = 83 calves). Fecal samples were collected via rectal palpation or from freshly voided, undisturbed fecal pats, weekly during the first 4 wk of the animal's life and then again at weaning. Eight total collections were made and 1,117 fecal samples cultured for Salmonella. One isolate from each culture-positive fecal sample was preserved for antimicrobial susceptibility screening and serotyping. Sixty-nine percent of the fecal samples were culture positive for Salmonella with no difference due to treatment (67.7 and 69% Salmonella positive for PWM and NPWM treatments, respectively). Few fecal samples (178/1,117; 15.9%) contained Salmonella concentrations above the limit of detection (～1 cfu/g of feces) with concentrations ranging from 1.0 to 6.46 cfu (log₁₀)/g of feces. Concentration was not affected by treatment. Seventeen different serotypes were identified, the majority of which were Montevideo and Anatum. A greater percentage of Typhimurium (87 vs. 13%), Muenchen (88 vs. 12%), and Derby (91 vs. 9%) were recovered from calves fed PWM compared with NPWM-fed calves. Conversely, Newport (12.5 vs. 86%), Bredeney (22.2 vs. 77.8%), and Muenster (12.5 vs. 87.5%) were lower in PWM compared with NPWM treatments. The majority (66.7%) of isolates were susceptible to all of the antibiotics examined. Results from this one commercial dairy suggest that milkborne Salmonella is not an important vector of transmission in dairy neonates, nor does pasteurization of waste milk influence fecal shedding of this pathogen. Caution should be used, however, when extrapolating results to other farms as Salmonella contamination of milk on farm is well documented. The potential benefits of pasteurization in disease prevention outweigh the potential risks of feeding a nonpasteurized product and warrants incorporation into any calf-rearing program using nonsaleable waste milk for feeding young dairy neonates.     

辐照和巴氏杀菌对番茄果浆品质及微生物生长情况的影响

为了探索辐照剂量和巴氏杀菌对番茄果浆的品质和杀菌效果的影响,对番茄果浆进行1、2、3kGy的辐照和巴氏杀菌处理,分别考察处理后的番茄果浆在25℃下的总酸、总糖、pH、花青素、VC含量及大肠菌群数、菌落总数和霉菌总数的变化情况,由此得到最适宜的处理方式.结果表明:辐照处理和巴氏杀菌处理对番茄果浆均起到了较好的杀菌效果,且...     

均质及杀菌条件对花生奶稳定性的影响

通过分析产品的稳定系数R，油脂析出率，离心沉淀率及粒度分布，研究均质及杀菌条件对花生奶稳定性的影响。结果表明，花生奶在温度65～70℃，压力25MPa下均质2次，再经121℃杀菌15min后产品的稳定性最佳。     

Possible implications of milk pasteurization on the manufacture and sensory quality of ripened cheese

Cheese made from raw milk represents an important proportion of the traditional cheeses, particularly in South European countries. Besides destruction of pathogenic bacteria, the most significant changes in milk relevant to cheesemaking, which are induced by pasteurization are:

• a partial elimination of the milk microorganisms which may grow in cheese during ripening,

• a partial or total activation or inhibition of the plasmin/plasminogen complex, cathepsin D, lipoprotein lipase and alkaline phosphatase. Enzymes from psychrotrophic bacteria, acid phosphatase and xanthine oxidase, which may be active during ripening, withstand pasteurization.,

• a slight (7%) denaturation of serum proteins and little or no modification of the cheesemaking properties (coagulation, acidification by lactic acid bacteria).

From experimental work carried out on several cheese varieties, comparing pasteurized or microfiltered milk and raw milk cheeses, it was found that facultatively heterofermentative lactobacilli, Micrococcaceae, enterococci, and propionibacteria in Swiss-type cheese, are found at higher levels in raw milk cheese. The main biochemical modification of cheese during ripening concerns the nature and extent of proteolysis. Although there is no clear trend in the breakdown of _s1- and β-caseins, milk pasteurization leads to a significant decrease of the amount of small peptides and free amino acids and to different HPLC profiles. Experiments carried out with sensory analysis show that, in all cases, pasteurized or microfiltered milk cheeses have received lower flavour intensity scores than raw milk cheeses. From this review, it is concluded that the indigenous milk microflora, with its diversity of species and strains, appears to be mainly responsible of the specific sensory properties of raw milk cheeses.     

巴氏杀菌对原料羊乳卫生质量的影响

本文就巴氏杀菌条件对原料羊乳和贮存过程中巴氏杀菌羊乳的菌落总数和大肠菌群变化规律进行研究。不同污染程度的原料羊乳分别在63℃/30min、72℃/30s和85℃/15s下处理,乳样中菌落总数和大肠菌群变化差异极为显著（p<0.05）。随着巴氏杀菌强度的增大,污染程度相同乳样的巴氏杀菌效率依次增高,污染程度不同乳样的巴氏杀菌效率却依次降低。巴氏杀菌后轻度污染和中度污染的乳样在5℃贮存7d后,其菌落总数和大肠菌群变化不显著（p>0.05）,均未超过巴氏杀菌乳国标GB19645-2010;重度污染的巴氏杀菌羊乳乳样在5℃贮存7d之后其菌落总数和大肠菌群已经显著超过巴氏杀菌乳国标GB19645-2010。因此原料羊乳在生产和贮存运输过程中要避免被微生物所污染,轻度和中度污染乳样的巴氏杀菌条件应选63℃/30min,贮存温度和时间应该控制在5℃/7d以内,以提高巴杀羊乳的品质和货架期。      

Effects of heating temperatures and addition of reconstituted milk on the heat indicators in milk

The objective of the present study was to evaluate the effect of heating temperatures and reconstituted milk on heat treatment indicators in milk by comparing the heat damage between raw milk and raw milk plus reconstituted milk (composite milk). The contents of lactulose, furosine, beta-lactoglobulin, and lactoperoxidase were determined after the heat indicators were heated to 65 to 115 °C for 15 s both in raw milk and composite milk. In the raw milk, the lactulose and furosine contents increased with increased heating temperature, while the beta-lactoglobulin content and lactoperoxidase activity decreased. The lactulose and furosine contents were increased after the addition of reconstituted milk. The reconstituted milk also significantly (P < 0.05) reduced the concentration of beta-lactoglobulin in the milk. Both heat treatment and an addition of reconstituted milk decreased the lactoperoxidase activity significantly (P < 0.05), and the lactoperoxidase activity was undetectable at 85 °C. The ratios of lactulose to furosine in pasteurized milk were higher than that in composite pasteurized milk. It is concluded that lactulose, furosine, and beta-lactoglobulin are suitable indicators of high heat pasteurization or raw milk, while lactoperoxidase may be used in monitoring mild heat pasteurization. Practical Application: Adequate heat treatment is necessary to destroy the microbes in raw milk. However, excessive heat treatment can result in inactivation of active compounds or loss of nutrients. The present study showed that the concentrations of lactulose, furosine, beta-lactoglobulin, and the activity of lactoperoxidase are sensitive to processing temperature and can serve as indicators of milk pasteurization.     

动力作用作为冷杀菌方法的可行性初探

通过鲜牛奶经纳米超高压均质机处理，取样，测定大肠菌群菌落数和细菌总数，结果表明，动力作用对牛乳中大肠菌群有致死效果；细菌总数最多可以降低3个对数单位，实验初步证实了动力杀菌作为一种冷杀菌的可行性。     

Efficacy of dairy on-farm high-temperature,short-time pasteurization of milk on the viability of Mycobacterium avium ssp. paratuberculosis

Feeding pasteurized milk to suckling calves is a popular practice used increasingly on dairy farms. Waste milk is frequently fed to calves because of its high nutritional value and economic benefits compared to milk replacement products. However, one of the disadvantages of feeding waste milk is the potential for exposure to a high number of bacterial contaminants, which may lead to serious illnesses or infections in calves. One of these contaminants is Mycobacterium avium ssp. paratuberculosis (MAP), the causative agent of Johne's disease (paratuberculosis). The transmission and distribution of paratuberculosis in dairy herds occurs mostly through the feeding newborn calves with contaminated colostrum or milk, because this age group is believed to be most susceptible to infection. To reduce the risk of transmission of pathogens, on-farm pasteurization of milk has become increasingly popular. In this study, we analyzed the efficacy of a new commercial high-temperature, short-time pasteurizer (73.5°C for 20 to 25 s) in terms of MAP inactivation under experimental on-farm conditions. The pasteurizer uses a newly developed steam-heating technique, allowing for the pasteurization of the transition milk without clumping. In 3 independent trials, we spiked fresh raw milk samples to a level of 10⁷ or 10⁴ viable MAP cells/mL before pasteurization. We examined the thermal inactivation and viability of MAP using culture and a D29 bacteriophage-based assay. To verify the identity and number of MAP cells, we also performed PCR assays. Pasteurization of the inoculated milk (10⁷ and 10⁴ MAP cells/mL) resulted in a remarkable reduction in viable MAP cells. The mean inactivation rate of MAP ranged from 0.82 to 2.65 log₁₀ plaque-forming units/mL, depending on the initial MAP amount inoculated and the addition of conservative agents to the pasteurized milk. Nevertheless, approximately 10³ MAP cells/mL remained viable and could be transferred to calves after high-temperature, short-time pasteurization of milk.     

A theoretical note on estimating the number of recoverable spores from survival curves having an ‘activation shoulder’

A ubiquitous feature of the survival curves of heat-treated Bacillus stearothermophilus and several other bacilli spores is the existence of a prominent ‘activation shoulder’. The shape of the survival curve alone, however, is insufficient to establish whether the observed initial increase in the number of ‘recoverable’ or ‘retrievable’ spores is due to heat activation, improved dispersion (‘de-clumping’) or both. But regardless of the mechanism, one can estimate a lower bound of the total recoverable spores using a crude empirical mathematical model. Estimates based on published data obtained with this model show that there can be 5–8 folds more recoverable spores in a heat-treated sample than the initial count would indicate, but this requires direct experimental verification. If correct, the model could serve as a tool to interpret the various possible shapes of ‘activation shoulders’ and to quantify the rate at which bacterial spores become ‘recoverable’.     

Invited review: The application of alkaline phosphatase assays for the validation of milk product pasteurization

Standard practices for indirectly assessing the pasteurization status of milk products are primarily based on the thermal inactivation kinetics of the endogenous milk enzyme, alkaline phosphatase (ALP). This assessment provides an invaluable, if not required, tool for both regulatory and in-house process control and validation. Endogenous milk ALP manifests a slightly higher heat resistance than the pathogenic microflora upon which pasteurization time and temperature requirements are based. Hence, ALP activity is recognized and accepted as the method of choice for the rapid validation of milk product pasteurization. However, ALP assays have notable limitations that must be understood if they are to be administered and interpreted correctly and the results are to be applied judiciously. Issues such as the reactivation of heat-denatured ALP and the presence of both heat-stable and -labile microbial ALP are addressed. A discussion of ALP in the milk of nonbovine species is presented based on the limited literature available. Some discussion of research involving alternative pasteurization indicators also is presented. This article is intended to summarize the pertinent details of the ALP assay for dairy products (noting the basis and limitations of various methods) and the processing, handling, and known compositional factors that influence the assay results.     

Use of hydrogen peroxide detection strips to determine the extent of pasteurization in whole milk

Commercially available hydrogen peroxide (H₂O₂) detection strips were shown to be effective in determining known levels of H₂O₂ in milks where the lactoperoxidase system (LPS) had been inactivated. In addition, the strips were evaluated for determination of residual H₂O₂ in LPS-activated milks, following a range of heat treatments. The detection of residual H₂O₂ corresponded with results of zero lactoperoxidase activity. Hence, the detection strips offer a simple and accurate method for detecting the absence of lactoperoxidase, and therefore can be used to determine whether the milk has been subjected to overpasteurization. The technique provides a more convenient method than the standard procedure based on oxidation of 1,4-phenylenediamine.     

原料乳生产过程中微生物污染的来源追溯研究

为研究原料乳微生物污染的来源,2011-2012年间,研究者对北京市原料乳产地的牛乳、牛体涂抹、挤乳设备涂抹等及生活用水、卧床土壤、饲料等进行了大量采样分析和调查,并针对采集样品中分离出的目的菌株进行了生化鉴定及分析.研究结果表明,该地区原料乳中的特征病原菌为金黄色葡萄球菌,对后期加工乳品质影响较大的微生物种类为嗜冷菌和芽孢杆菌属.在此基础上,研究者对原料乳和其他来源样品中的这三种菌利用16S rRNA分子生物学技术建立了目的菌株来源系统发育树,以此来进行基因序列比对分析,确定了原料乳中这三种菌的主要来源,为有效减少和控制该地区原料乳污染提供了参考.     

Effect of different colored filters on photooxidation in pasteurized milk

The effect of different colored filters and atmospheres on photooxidation and quality in milk was studied. Pasteurized bovine milk (3.9% fat) was packed in 2 different atmospheres (air and N₂) and exposed to light for 20 h at 4°C under 8 transparent filters with different light transmission properties. The following transparent, noncolored, and colored filters based on polyethylene terephthalate (PET) were used: noncolored (PET), noncolored with 2 different UV-block regions, yellow, green, amber, orange, and red. Control samples were stored in darkness and in a carton. Sensory evaluation showed off flavors significantly increased in milk stored under all filters compared with the control samples. Variation in atmosphere resulted in significant differences in formation of rancid flavor in milk stored under different filters. Milk samples stored in N₂ underwent the most sensory deterioration under orange and red filters, whereas milk samples stored in air were most deteriorated under noncolored filters. According to the oxidation compounds measured by gas chromatography, milk samples stored under noncolored and orange filters were highly oxidized, whereas red, green, and amber filters offered better protection against photooxidation. Fluorescence spectroscopy was used to examine the degradation of photosensitizers (riboflavin, protoporphyrin, and chlorophyllic compounds) in the milk samples. Degradation of protoporphyrin and chlorophyllic compounds in N₂ correlated well with sensory properties related to photooxidation (R² = 0.75-0.95). The study indicates that protoporphyrin and chlorophyllic compounds were effective photosensitizers in milk. To avoid photooxidation in milk, it is therefore important to protect it against light from the UV spectrum as well as light from the entire visible region.