Survival of Mycobacterium paratuberculosis and preservation of immunoglobulin G in bovine colostrum under experimental conditions simulating pasteurization

OBJECTIVE: To determine whether Mycobacterium paratuberculosis could survive in colostrum after pasteurization. Additionally, this study investigated the effect pasteurization had on IgG concentration in colostrum. ANIMALS: Colostrum samples were collected from cattle (beef and dairy) owned by the state of Ohio. PROCEDURE: Colostrum was divided into aliquots and inoculated with variable concentrations of M paratuberculosis (ATCC No. 19698: 10(4), 10(3), and 10(2) colony-forming units/ml). Half the samples at each concentration were subjected to pasteurization temperatures (63 C) for 30 minutes and the remainder were kept at approximately 20 to 23 C. All samples were incubated (Herrold's egg yolk medium with and without mycobactin J) and observed for growth during the next 16 weeks. Additionally, the IgG concentration of colostrum was determined by radioimmunoassay before and after pasteurization. Samples that coagulated at pasteurization temperatures were mechanically resuspended before measurement of IgG concentration. RESULTS: Growth of M paratuberculosis was retarded but not eliminated by pasteurization. Growth was observed in all unpasteurized samples incubated on Herrold's egg yolk medium with mycobactin J but in only 2 of 18 pasteurized samples similarly cultured. Growth from pasteurized samples appeared 5 to 9 weeks after growth was observed from nonpasteurized samples. Mean colostral IgG concentration was 44.4 g/L in nonpasteurized samples and 37.2 g/L in pasteurized samples, a decrease of 12.3%. High-quality colostrum (> 48 g of IgG/L) had a significantly greater loss of IgG concentration than did colostrum of lesser quality (P = 0.002). CONCLUSIONS: Pasteurization lessened, but did not eliminate, growth of M paratuberculosis from experimentally inoculated colostrum samples. Pasteurization resulted in a significant decrease in colostral IgG concentration but not to an unmanageable level that would preclude the colostrum's use for passive transfer of immunity. CLINICAL RELEVANCE: Colostrum is macrophage rich and may serve as a source of M paratuberculosis infection to calves. Pasteurization of colostrum may lessen the risk of infection, but will not totally eliminate M paratuberculosis.     

Thermal inactivation of Mycobacterium paratuberculosis in milk

Thermal inactivation of Mycobacterium paratuberculosis, a suspected human pathogen, was determined in ultrahigh-temperature whole milk. Three strains of M. paratuberculosis were examined for survival at temperatures from 55 to 75 degrees C using a submerged glass capillary tube method. Clumped and declumped suspensions of the cultures were used to determine the rate of heat inactivation and survival at pasteurization temperatures. Methods for declumping M. paratuberculosis included the use of glass beads, vortexing, and passing the cells through a 26-gauge needle. The latter procedure was found to be superior over other methods and did not affect the viability of cells. Capillary tubes filled with milk containing 4 x 10(6) to 3 x 10(7) CFU/ml were heated at temperatures ranging from 55 to 75 degrees C. At 55 degrees C, minimal thermal inactivation was observed for clumped and declumped cells. At 58 degrees C, thermal inactivation ranging from 0.3 to 0.7 log reduction was observed for both clumped and declumped suspensions. D values at 60 degrees C ranged from 8.6 to 11 min and 8.2 to 14.1 min for clumped and declumped cells, respectively. At 63 degrees C, the D values ranged from 2.7 to 2.9 and 1.6 to 2.5 min for clumped and declumped cells, respectively. Survival of M. paratuberculosis at initial levels ranging from 44 to 10(5) CFU/ml at pasteurization treatment (63 degrees C for 30 min and 72 degrees C for 15 s) was also determined. No survivors were observed after incubating plates for up to 4 months on Middlebrook 7H11 agar and up to 2 months on Herrold's egg yolk medium. The sensitivity of the plating method was 1 CFU/250 microliters. These results demonstrate that low levels of M. paratuberculosis, as might be found in raw milk, will not survive pasteurization treatments.     

Effect of pasteurization on infectivity of Cryptosporidium parvum oocysts in water and milk

Cryptosporidium parvum is a major cause of diarrheal disease in humans and has been identified in 78 other species of mammals. The oocyst stage, excreted in feces of infected humans and animals, has been responsible for recent waterborne outbreaks of human cryptosporidiosis. High temperature and long exposure time have been shown to render oocysts (suspended in water) noninfectious, but for practical purposes, it is important to know if high-temperature--short-time conditions (71.7 degrees C for 15 s) used in commercial pasteurization are sufficient to destroy infectivity of oocysts. In this study, oocysts were suspended in either water or whole milk and heated to 71.7 degrees C for 15, 10, or 5 s in a laboratory-scale pasteurizer. Pasteurized and nonpasteurized (control) oocysts were then tested for the ability to infect infant mice. No mice (0 of 177) given 10(5) oocysts pasteurized for 15, 10, or 5 s in either water or milk were found to be infected with C. parvum on the basis of histologic examination of the terminal ileum. In contrast, all (80 of 80) control mice given nonpasteurized oocysts were heavily infected. These data indicate that high-temperature--short-time pasteurization is sufficient to destroy the infectivity of C. parvum oocysts in water and milk.     

Gamma-glutamyltransferase as a marker for the pasteurization of raw milk

The degree and rate of inactivation of gamma-glutamyltransferase in raw cow's milk by heating at 50, 60, 70, and 80 degrees C for 1, 2, 3, 5, 10, 15, 20, 25, and 30 min were measured to evaluate the suitability of this enzyme as a marker for the pasteurization of milk. The enzymes alkaline phosphatase and lactate dehydrogenase were also measured under similar conditions for comparison. The patterns of heat inactivation of gamma-glutamyltransferase and alkaline phosphatase were similar, with only a minimal inactivation of the enzymes at 50 degrees C. The rate of inactivation increased as a result of increasing temperatures and time. A complete inactivation of both enzymes was seen at 70 degrees C after 10 min and at 80 degrees C after 1 min. Lactate dehydrogenase showed a higher heat resistance with almost complete inactivation at 70 degrees C for 30 min, and compete inactivation at 80 degrees C for 3 min. No activities of these enzymes were found in commercially pasteurized or heat-treated milk. The levels of gamma-glutamyltransferase in raw milk were between 8 and 10% higher than those of alkaline phosphatase and lactate dehydrogenase, making it more sensitive and accurate as a testing marker. It seems that gamma-glutamyltransferase may serve as a good pasteurization marker. Furthermore, the simplicity of testing and the availability of commercial kits for testing by both wet and dry chemistry make it an attractive choice, especially because dry chemistry procedures overcome the difficulties originating from the turbidity of milk, which interferes with spectrophotometric procedures.     

Automated control and monitoring of thermal processing using high temperature, short time pasteurization

High temperature, short time pasteurization was used to evaluate a computer-based system for controlling the pasteurization process, acquiring data, and monitoring records. Software was used for the control of hot water temperature, flow rate through the centrifugal timing pump, and diversion of under-processed product. Three types of control strategies were conducted: single loop, cascade, and multivariable. The single loop control strategy showed the most rapid responses to temperature changes, but the temperature response curve was slowest to return to its set point. The cascade control strategy showed slower recoveries to temperature changes, but the temperature response curve was smoother. The multivariable control strategy responded slightly faster than the cascade control strategy, and the temperature response curve was slightly smoother than the cascade control strategy. The multivariable control strategy was able to control the flow diversion valve by the use of a lethality controller. The data acquisition system, used to monitor the data obtained from the high temperature, short-time pasteurization system, was within +/- 0.1 degree C of the temperature recorded by the safety thermal limit recorder. Reliability was determined by examining the changes in the position of the flow diversion valve to identify process deviations and by comparing the changes to the event marker on circular charts. The data acquisition system was an effective alternative for monitoring the completeness of data.     

Food poisoning. Causes, remedies, and prevention

Food producers and consumers must continue to take precautions against foodborne diseases, and early diagnosis and appropriate treatment of these illnesses are essential. Food products and water can become contaminated with microorganisms and toxins that make people ill, and the very young, the elderly, and immunocompromised individuals are especially susceptible. Education of healthcare providers, food handlers, and the public is critical in reducing the incidence and spread of foodborne illness. Changes in eating habits and lifestyle and increased availability of both domestic and imported foods have made food hazards a more complex public health issue. Although most foodborne illnesses can be avoided by safe food handling procedures (see box on page 134), risk reduction is very important at every step from source to table. A concerted effort is needed to ensure continuing safety of the food supply in the United States while also assuring access to a wide variety of healthful foods. Time will tell whether consumers will accept irradiation of meats. If accepted, irradiation could rank in importance with pasteurization of milk and chlorination of water as a public health measure.     

Mycobacterium paratuberculosis: a potential food-borne pathogen?

Mycobacterium paratuberculosis commonly infects dairy cattle, leading to Johne's disease, which is also known as paratuberculosis. The infection is chronic progressive, and incurable. As the infection progresses, excretion of M. paratuberculosis in feces and milk occurs, and the bacterium spreads through the blood to multiple internal organs. Consequently, raw products originating from cattle may harbor M. paratuberculosis. Thermal treatments, such as pasteurization, are commonly relied on to kill food-borne bacterial pathogens that can infect humans. The small number of studies conducted to determine the thermal resistance of M. paratuberculosis suggest that it is less susceptible to destruction by heat killing than are milkborne zoonotic bacterial pathogens such as Listeria spp. or Mycobacterium bovis. Published reports concerning the thermal resistance of M. paratuberculosis in milk are reviewed herein, and key issues concerning the efficacy of pasteurization for elimination of M. paratuberculosis from milk are summarized.     

Virus inactivation and prevalence of GBV-C in haemophiliacs

A new putative hepatitis virus has recently been discovered and termed GB virus C (GBV-C). We investigated the prevalence of this virus among 50 haemophiliacs treated with non-virus-inactivated clotting factor concentrates prior to 1985 and 21 haemophiliacs treated exclusively with virus-inactivated clotting factor concentrates. In the first group the prevalence of GBV-C based on PCR and ELISA was 46%. In the second group the prevalence of GBV-C was similar to that of healthy blood donors (5%). We therefore conclude that GBV-C is reliably inactivated by modern virucidal methods such as vapour heating, pasteurization and treatment with solvent/detergent mixtures.     

A novel esterase from Saccharomyces carlsbergensis, a possible function for the yeast TIP1 gene

An extracellular esterase was isolated from the brewer's yeast, Saccharomyces carlsbergensis. Inhibition by diisopropyl fluorophosphate shows that the enzyme has a serine active site. By mass spectrometry, the molecular weight of the enzyme was 16.9 kDa. The optimal pH for activity was in the range of four to five. Esterase activity was found in beer before pasteurization, and a low level of activity was still present after pasteurization. Caprylic acid, which is present in beer, competitively inhibited the esterase. The substrate preference towards esters of p-nitrophenol indicated that the enzyme prefers esters of fatty acids from four to 16 carbon atoms. The esterase has lipolytical activity; olive oil (C-18:1), which is a classical substrate for lipase, was hydrolysed. N-terminal sequence analysis of the esterase yielded a sequence which was identical to the deduced amino acid sequence of the S. cerevisiae TIP1 gene. The esterase preparation did not appear to contain significant amounts of other proteins than Tip1p, indicating that the TIP1 gene is the structural gene for the esterase.     

Bioretention Impact on Runoff Temperature in Trout Sensitive Waters

A study was conducted in western North Carolina, along the southeastern extent of the U.S. trout populations, to examine the effect of bioretention areas on runoff temperature. Four bioretention areas were monitored during the summers of 2006 and 2007. It was found that smaller bioretention areas, with respect to the size of their contributing watershed, were able to significantly reduce both maximum and median water temperatures between the inlet and outlet. The proportionately larger bioretention areas were only able to significantly reduce maximum water temperatures between the inlet and outlet; however, these systems showed evidence of substantial reductions in outflow quantity, effectively reducing the thermal impact. Despite temperature reductions, effluent temperatures still posed a potential threat to coldwater streams during the peak summer months. During the summer months, effluent temperatures were generally coolest at the greatest soil depths, supporting evidence of an optimum drain depth between 90 and 120 cm. The ability of bioretention areas to reduce storm-water temperature and flows supports their application to reduce the thermal impacts of urban storm-water runoff.     

堆存温度对半水磷石膏胶凝性能影响

半水磷石膏（HPG）长时间堆存状态下会出现固结现象,其胶凝性能也相应下降。以室内HPG结晶水检测和单轴压缩试验为基础,通过设定4种不同堆存温度,分别为20,40,60和80 ℃,探究不同堆存温度作用下HPG试样结晶水质量分数变化和堆存后制备的充填胶凝材料（HCM）抗压强度发展规律,并采用扫描电镜等微观分析手段研究堆存温度对其强度影响机制。结果表明,堆存温度对HPG胶凝性能影响显著,高的堆存温度会加快HPG试样中的自由水转变为结晶水速率,而且会抑制堆存后制备的HCM强度发展。采用数据标准化对不同堆存温度作用后的试样抗压强度作出预测,被证实与实测值较吻合。微观分析发现,堆存温度主要影响体系的过饱和度,而使不同堆存温度作用后制备的HCM微观形态表现差异。      

Comparison of responses of men to immersion in circulating water at 40.0 and 41.5 degrees C

BACKGROUND: The recommended maximum water temperature for public hot tubs has been set at 40.0 degrees C, but no research has been published on human immersion in hot water at higher temperatures. HYPOTHESIS: We hypothesized that thermoregulatory and cardiovascular responses at two water temperatures would be proportional to the water:blood temperature gradients. METHODS: Six healthy men were immersed for 21 min in circulating hot water at 40.0 and 41.5 degrees C in separate trials in random order 1-3 wk apart. Measurements included heart rate, systolic BP, esophageal, rectal, and non-immersed skin temperatures, sweat rate, and perceived comfort. RESULTS: The rise in all body temperatures, sweat rate, and heart rate were significantly greater in the 41.5 vs. 40.0 degrees C water. Peak esophageal temperatures were 38.3 +/- 0.2 degrees C vs. 37.8 +/- 0.03 degrees C, peak sweat rates were 0.48 +/- 0.05 vs. 0.32 +/- 0.03 kg x m(-2) x h(-1), and peak heart rates were 123 +/- 7 vs. 108 +/- 5 bpm, respectively. Systolic BPs followed different patterns of response in each trial, whereas diastolic pressures were not different between trials. Comfort at each level of immersion was reduced during the 41.5 degrees C trial compared with the 40.0 degrees C in excess of that predicted by difference in esophageal temperature between the trials. CONCLUSIONS: These results suggest that risks of hyperthermia or adverse cardiovascular effects in hot tubs may not be greater in water above 40.0 degrees C unless perceptual judgment is impaired. Hypotension when standing to exit the tub occurred in both trials and may represent a potential hazard to hot tub use.     

板坯动态二冷增量型PID控制研究

为解决连铸拉速工况变化时因使用一级水表配水控制所引起的铸坯表面温度波动的问题,建立了考虑铸坯纵向导热的板坯传热瞬态模型.通过Gleeble试验获得钢的热塑性,制定了铸坯的目标温度.采用增量型PID控制方法开发了动态二冷控制软件Visual Cast.结果表明:与传统的一级水表相比,当拉速发生变化时VisualCast能...     

Superionic and metallic states of water and ammonia at giant planet conditions

The phase diagrams of water and ammonia were determined by constant pressure ab initio molecular dynamic simulations at pressures (30 to 300 gigapascal) and temperatures (300 to 7000 kelvin) of relevance for the middle ice layers of the giant planets Neptune and Uranus. Along the planetary isentrope water and ammonia behave as fully dissociated ionic, electronically insulating fluid phases, which turn metallic at temperatures exceeding 7000 kelvin for water and 5500 kelvin for ammonia. At lower temperatures, the phase diagrams of water and ammonia exhibit a superionic solid phase between the solid and the ionic liquid. These simulations improve our understanding of the properties of the middle ice layers of Neptune and Uranus.     

Physical and mathematical models of steel flow and heat transfer in a tundish heated by plasma

Temperature control of liquid steel by plasma heating is physically and mathematically modeled. A dimensionless plasma heating number is employed for scaling up of heating operations between a steam jet for water model and a plasma prototype. Overall responses of step-input temperatures in steel are fairly well predicted by the physical model. Fluid-flow structure and thermal fields, in terms of dimensionless temperatures, of steel are different from those of water. Two positions of plasma in a tundish were studied, centered and off-centered. The second position decreased the heating response of a plasma, although a more homogeneous thermal field was obtained in regard to the first position. Consequently, a centered position offered a faster thermal response than the off-centered one. Flow controllers decreased the efficiency of plasma due to the formation of stagnant zones located in their proximity, which were not active enough to exchange momentum and heat with the bulk flow.     

Experimental study of boiling heat transfer during subcooled water jet impingement on flat steel surface

Abstract

The growth in demand for high quality metal alloys has placed considerable emphasis on the type of cooling methods used in manufacturing processes, in particular, the production of highly tailored steel through controlled cooling on the runout table. The present study focuses on the heat transfer (cooling of hot rolled steel strips) on a runout table. The purpose of the study was to develop an efficient experimental method and collect temperature data under conditions similar to those that occur during industrial runout table conditions in a steelmill. Surface and internal temperatures were measured during transient cooling of a flat, upward facing fixed steel plate cooled by a highly subcooled single, circular, free surface jet of water. Measurements were made at stagnation and several streamwise distances from the stagnation point. A numerical, finite difference model was applied to calculate the surface heat flux using measured temperatures. The effect of water flowrate and subcooling on the overall heat transfer with emphasis on the maximum heat flux is discussed.     

Solar disinfection of drinking water contained in transparent plastic bottles: characterizing the bacterial inactivation process

A series of experiments is reported to identify and characterize the inactivation process in operation when drinking water, heavily contaminated with a Kenyan isolate of Escherichia coli, is stored in transparent plastic bottles that are then exposed to sunlight. The roles of optical and thermal inactivation mechanisms are studied in detail by simulating conditions of optical irradiance, water turbidity and temperature, which were recorded during a series of solar disinfection measurements carried out in the Kenyan Rift Valley. Optical inactivation effects are observed even in highly turbid water (200 ntu) and at low irradiances of only 10 mW cm-2. Thermal inactivation is found to be important only at water temperatures above 45 degrees C, at which point strong synergy between optical and thermal inactivation processes is observed. The results confirm that, where strong sunshine is available, solar disinfection of drinking water is an effective, low cost method for improving water quality and may be of particular use to refugee camps in disaster areas. Strategies for improving bacterial inactivation are discussed.     

Ovarian development and parity determination in Culiseta melanura (Diptera: Culicidae)

Eastern equine encephalomyelitis virus (EEE) is perpetuated in a maintenance cycle that involves Culiseta melanura (Coquillett) as the enzootic vector and passerine birds as the amplifying hosts. Amplification of virus in any given year requires an influx of nulliparous Cs. melanura in the presence of susceptible avian hosts. We conducted laboratory experiments at constant temperatures from 10 to 34 degrees C to develop thermal heat summation models to predict emergence in nature. Embryonic development progresses slowly at 10 degrees C, and the time to eclosion decreased significantly as temperatures increased to 28 degrees C. High temperature were lethal and eggs failed to hatch at 32 degrees C. The thermal minimum (t0) for embryonic development was 9.38 degrees C, and 38.46 degrees-days (DD) were required for egg hatch. The time for larval development decreased with increasing temperatures. Cs. melanura larvae develop in subterranean habitats (crypts) where water temperatures remain below 20 degrees C throughout the summer. Under controlled conditions, egg hatch to emergence took 8 mo at 10 degrees C, 3 mo at 16 degrees C, and 1 mo at 22 degrees C. The thermal minimum for larval development (t0) was 8.5 degrees C, and 467.29 DD were required from eclosion to adult emergence. Our findings indicate that Cs. melanura is well suited to develop in cold water crypts where the larvae are collected most frequently. The mosquito appears to be bivoltine in the northeast with an overwintering generation of larvae that emerges as a spring brood of adults and a summer generation of larvae that emerges in fall. Higher than normal water temperatures hasten development of the summer generation and increase the probability for amplification of EEE by bringing large numbers of nulliparous mosquitoes into contact with recrudescing virus for subsequent transfer to the growing population of susceptible juveniles. Water temperatures in the crypts also may regulate the northern limit for virus amplification each year.     

Thermal denaturation of ribonuclease A characterized by water 17O and 2H magnetic relaxation dispersion

Water oxygen-17 and deuteron nuclear magnetic relaxation dispersion (NMRD) measurements were used to characterize ribonuclease A (RNase A) in the course of thermal denaturation at pH 2 and 4. The structure and dynamics of the protein were probed by specific long-lived water molecules, by the short-lived surface hydration, and by labile side-chain hydrogens. The NMRD data show that native RNase A contains at least three water molecules with a mean residence time of 8 ns at 27 degreesC and an activation enthalpy of ca. 40 kJ mol-1. These water molecules are identified with some or all of six ordered water molecules partly buried in surface pockets in the crystal structure of RNase A. The loss of the 17O dispersion at higher temperatures demonstrates that, in the thermally denatured protein, these surface pockets are either not present or undergoing large structural fluctuations on a subnanosecond time scale. The relaxation dispersion step vanishes monotonically and essentially in concert with the CD denaturation curves, thus ruling out the existence of equilibrium intermediates with a substantial amount of non-native and long-lived hydration water. The NMRD data show that thermally denatured RNase A has a relatively compact but highly flexible structure. The global solvent exposure and the hydrodynamic volume of the denatured protein are much less than for maximally unfolded disulfide-intact RNase A. The NMRD data show that thermal denaturation is accompanied by a large reduction of the mean-square orientational order parameter of side-chain O-H bonds, implying that, in the denatured state, these side chains sample a wide distribution of conformational states on a subnanosecond time scale.