Predictive model for inactivation of Campylobacter spp. by heat and high hydrostatic pressure

Campylobacter represents one of the leading causes of foodborne enteritis. Poultry and its products frequently transmit the pathogen. The objective of the present study was to model predictively the short-term inactivation of Campylobacter in a ready-to-eat poultry product to develop an economic high-pressure treatment. We inactivated baroresistant strains of Campylobacter jejuni and Campylobacter coli, grown to stationary phase on nutrient agar and inoculated in poultry meat slurry, by heat and high hydrostatic pressure. Incubation at ambient pressure at 70 degrees C for 1 min and at 450 MPa at 15 degrees C for 30 s inactivated more than 6 log CFU of this foodborne pathogen per ml of poultry meat slurry. Thermal and pressure inactivation kinetics of C. coli and C. jejuni in poultry meat slurry were accurately described by a first-order kinetic model. A mathematical model was developed from 10 to 65 degrees C and from ambient to 500 MPa that predicts the reduction in numbers of Campylobacter in response to the combination of temperature, pressure, and treatment time. We suggest the high-pressure treatment of foods to avoid health risks caused by Campylobacter. The nonthermal short-term treatment of the examined food model system represents a successful step to an economic high-pressure procedure.     

Microassay for measuring thermal inactivation of H5N1 high pathogenicity avian influenza virus in naturally infected chicken meat

A precise, reproducible microassay was developed to measure thermal inactivation of high pathogenicity avian influenza (HPAI) virus in chicken meat. Small pieces of breast or thigh meat (0.05 g) from chickens infected with A/chicken/Pennsylvania/1370/1983 (H5N2) (PA/83) or A/chicken/Korea/ES/2003 (H5N1) (Korea/03) HPAI viruses were tested for inactivation in the heating block of a thermocycler. Korea/03 infected thigh and breast meat had higher virus concentrations (10(6.8) and 10(5.6) mean embryo infectious doses [EID(50)]/g, respectively) compared to PA/83 infected thigh and breast meat (10(2.8) and 10(2.3) EID(50)/g, respectively). The samples were ran through a ramp-up cycle from 25 to 70 degrees C, and meat samples were removed and examined for virus infectivity at 30, 40, 50, 60 and 70 degrees C, and after treatment for 1, 5, 10, 30 and 60 s at 70 degrees C. The reduction in virus infectivity titers was dependent on virus concentration and no HPAI virus was isolated after 1 s of treatment at 70 degrees C. A change in coloration from pink-tan to white was associated with a loss in recovery of infectious virus. The microassay provided a predictable and reproducible method to measure thermal inactivation of HPAI virus in chicken meat.     

Thermal inactivation of avian influenza and Newcastle disease viruses in chicken meat

Avian influenza viruses (AIV) and Newcastle disease viruses (NDV) of high pathogenicity cause severe systemic disease with high mortality in chickens and can be isolated from the meat of infected chickens. Although AIV and NDV strains of low pathogenicity are typically not present in chicken meat, virus particles in respiratory secretions or feces are possible sources of carcass contamination. Because spread of AIV and NDV is associated with movement of infected birds or their products, the presence of these viruses in chicken meat is cause for concern. This study presents thermal inactivation data for two viruses of high pathogenicity in chickens (AIV strain A/chicken/Pennsylvania/1370/1983 and NDV strain APMV-1/ chicken/California/S0212676/2002) and two viruses of low pathogenicity in chickens (AIV strain A/chicken/Texas/298313/ 2004 and NDV strain APMV-1/chicken/Northern Ireland/Ulster/1967). Under the conditions of the assay, high-pathogenicity AIV was inactivated more slowly in meat from naturally infected chickens than in artificially infected chicken meat with a similar virus titer. In contrast, high-pathogenicity NDV was inactivated similarly in naturally and artificially infected meat. Linear regression models predicted that the current U.S. Department of Agriculture-Food Safety and Inspection Service time-temperature guidelines for cooking chicken meat to achieve a 7-log reduction of Salmonella also would effectively inactivate the AIV and NDV strains tested. Experimentally, the AIV and NDV strains used in this study (and the previously studied H5N1 high-pathogenicity AIV strain A/chicken/Korea/ES/2003) were effectively inactivated in chicken meat held at 70 or 73.9 degrees C for less than 1 s.     

Effect of high hydrostatic pressure processing on freely suspended and bivalve-associated T7 bacteriophage

The effectiveness of hydrostatic pressure processing (HPP) for inactivating viruses has been evaluated in only a limited number of studies, and most of the work has been performed with viruses freely suspended in distilled water. In this work, HPP inactivation of freely suspended and shellfish-associated bacteriophage T7 was studied. T7 was selected in hopes that it could serve as a model for animal virus behavior. Clams (Mercenaria mercenaria) and oysters (Crassostrea virginica) were homogeneously blended separately and inoculated with bacteriophage T7. The inoculated bivalve meat and the freely suspended virus samples were subjected to HPP under the following conditions: 2, 4, and 6 min at 241.3, 275.8, and 344.7 MPa pressure and temperatures of 29.4 to 35, 37.8 to 43.3, and 46.1 to 51.7 degrees C. Reductions of 7.8 log PFU (100% inactivation) were achieved for freely suspended T7 at 344.7 MPa for 2 min at 37.8 to 43.3 degrees C. At 46.1 to 51.7 degrees C, T7 associated with either clams or oysters was inactivated at nearly 100% (>4 log PFU) at all pressure levels and durations tested. These results indicate that T7 is readily inactivated by HPP under the proper conditions, may be made more susceptible to HPP by mixing with shellfish meat, and may serve as a viable model for the response of several animal viruses to HPP.     

蛋类禽流感病毒检测报告

了解丽水市禽蛋标本禽流感病毒的感染状况。方法 对随机抽样的690只鸡、鸭、鸽蛋及126份农贸市场外环境对照标本采用荧光定量PCR方法检测禽流感病毒H7N9、H5和H9亚型核酸。结果 690只禽蛋标本H7N9、H5及H9亚型禽流感病毒核酸检测均为阴性,126份外环境对照标本H7N9亚型禽流感病毒核酸检测均为阴性,H5和H9亚型阳性标本各2份,阳性率为3.17%。结论 禽蛋中H7N9、H5及H9亚型禽流感病毒污染率低,但丽水市禽间存在H5和H9亚型禽流感的传播。各有关部门应加强合作,采取切实有效措施,防止人感染禽流感。     

Effect of small, acid-soluble proteins on spore resistance and germination under a combination of pressure and heat treatment

To find the range of pressure required for effective high-pressure inactivation of bacterial spores and to investigate the role of alpha/beta-type small, acid-soluble proteins (SASP) in spores under pressure treatment, mild heat was combined with pressure (room temperature to 65 degrees C and 100 to 500 MPa) and applied to wild-type and SASP-alpha-/beta- Bacillus subtilis spores. On the one hand, more than 4 log units of wild-type spores were reduced after pressurization at 100 to 500 MPa and 65 degrees C. On the other hand, the number of surviving mutant spores decreased by 2 log units at 100 MPa and by more than 5 log units at 500 MPa. At 500 MPa and 65 degrees C, both wild-type and mutant spore survivor counts were reduced by 5 log units. Interestingly, pressures of 100, 200, and 300 MPa at 65 degrees C inactivated wild-type SASP-alpha+/beta+ spores more than mutant SASP-alpha-/beta- spores, and this was attributed to less pressure-induced germination in SASP-alpha-/beta- spores than in wild-type SASP-alpha+/beta+ spores. However, there was no difference in the pressure resistance between SASP-alpha+/beta+ and SASP-alpha-/beta- spores at 100 MPa and ambient temperature (approximately 22 degrees C) for 30 min. A combination of high pressure and high temperature is very effective for inducing spore germination, and then inactivation of the germinated spore occurs because of the heat treatment. This study showed that alpha/beta-type SASP play a role in spore inactivation by increasing spore germination under 100 to 300 MPa at high temperature.     

Temperature-assisted pressure inactivation of Listeria monocytogenes in turkey breast meat

Ready-to-eat turkey breast meat samples were surface-inoculated with a five-strain cocktail of Listeria monocytogenes cultures to a final concentration of approximately 10(7) CFU/g. The inoculated meat samples were vacuum-packaged and pressure treated at 300 MPa for 2 min, 400 MPa for 1 min, and 500 MPa for 1 min at initial sample temperatures of 1, 10, 20, 30, 40, 50, and 55 degrees C. L. monocytogenes was most resistant to pressure at temperatures between 10 and 30 degrees C. As temperature decreased below 10 degrees C or increased over 30 degrees C, its pressure sensitivity increased. This enhanced inactivation effect was more pronounced when meat samples were treated at higher temperature than at lower temperature. For example, a 1-min treatment of 500 MPa at 40 degrees C reduced the counts by 3.8 log(10), while at 1 and 20 degrees C the same treatment reduced counts by 1.4 and 0.9 log(10), respectively (P<0.05). The survival curves of L. monocytogenes were obtained at 300 MPa and 55 degrees C, 400 MPa and 50 degrees C, and 500 MPa and 40 degrees C. With increasing treatment time, the three survival curves showed a rapid initial drop in bacteria counts with a diminishing inactivation rate or tailing effect. The survival data were fitted with a linear and a nonlinear, Weibull, models. The Weibull model consistently produced better fit to the survival data than the linear model.     

Inactivation of Escherichia coli O157:H7 in orange juice using a combination of high pressure and mild heat

The effect of high pressure on the survival of a pressure-resistant strain of Escherichia coli O157:H7 (NCTC 12079) in orange juice was investigated over the pH range 3.4 to 5.0. The pH of commercial, sterile orange juice was adjusted to 3.4, 3.6, 3.9, 4.5, or 5.0. The juice was then inoculated with 10(8) CFU ml(-1) of E. coli O157:H7. The inoculated orange juice was subjected to pressure treatments of 400, 500, or 550 MPa at 20 degrees C or 30 degrees C to determine the conditions that would give a 6-log10 inactivation of E. coli O157:H7. A pressure treatment of 550 MPa for 5 min at 20 degrees C produced this level of kill at pH 3.4, 3.6, 3.9, and 4.5 but not at pH 5.0. Combining pressure treatment with mild heat (30 degrees C) did result in a 6-log10 inactivation at pH 5.0. Thus, the processing conditions (temperature and time) must be considered when pressure-treating orange juice to ensure microbiological safety.     

Response of four types of coliphages to high hydrostatic pressure

Pressure inactivation of four types of coliphages, varphiX 174 (ssDNA virus), MS2 (ssRNA virus), lambda imm434 (dsDNA virus) and T4 (dsDNA virus), was studied to evaluate their potential as human enteric viral surrogates for use in validation of commercial pressure processing treatments. Phage varphiX 174 demonstrated an unexpected high resistance to pressure with no more than 1-log(10) reduction observed following exposures to 350-600 MPa. There was no greater than 1-log(10) reduction below 500 MPa for MS2 in modified phosphate-buffered saline, but a 3.3-log(10) reduction was observed for MS2 pressurized at 600 MPa. Coliphages lambda imm434 and T4 were relatively sensitive to pressure in demonstrating inactivation at 350 MPa. At 21 degrees C, lambda imm434 was inactivated in modified phosphate-buffered saline or Dulbecco's Modified Eagle's Medium plus 5% fetal bovine sera by at least 7.5-log(10) when exposed to 400 MPa for 5 min. Treatment at 450 MPa for 5 min was necessary to obtain a log(10) reduction of 6-7 for T4.     

A Chromatographic Strip for Rapid Semi-quantitative Detection of H5 Subtype Avian Influenza Viruses in Poultry

An immunochromatographic strip was developed for rapid semi-quantitative of the H5 subtype of avian influenza viruses (H5 AIVs) in poultry. A specific monoclonal antibody 1C4 for H5 AIV hemagglutinin (HA) was produced and conjugated with colloidal gold as the detecting probes for the immunochromatographic strips, and a polyclonal antibody against H5N1 was used to firm the test line. Different from the traditional strips, four test lines with a same capture complex of different concentrations were designed on the nitrocellulose membrane. By contrast with the HA tests and the simulated infection experiment, the immune strip was proved to be specific for the detection of H5 AIV within 10 min and had the character to reflect the viruses infection degree in different organs of the infected poultry. It should be useful for diagnosing and semi-quantitative analysis of the H5 subtype of AIV.     

Thermal inactivation of H5N1 high pathogenicity avian influenza virus in naturally infected chicken meat

Thermal inactivation of the H5N1 high pathogenicity avian influenza (HPAI) virus strain A/chicken/Korea/ES/2003 (Korea/03) was quantitatively measured in thigh and breast meat harvested from infected chickens. The Korea/03 titers were recorded as the mean embryo infectious dose (EID50) and were 10(8.0) EID50/g in uncooked thigh samples and 10(7.5) EID50/g in uncooked breast samples. Survival curves were constructed for Korea/03 in chicken thigh and breast meat at 1 degrees C intervals for temperatures of 57 to 61 degrees C. Although some curves had a slightly biphasic shape, a linear model provided a fair-to-good fit at all temperatures, with R2 values of 0.85 to 0.93. Stepwise linear regression revealed that meat type did not contribute significantly to the regression model and generated a single linear regression equation for z-value calculations and D-value predictions for Korea/03 in both meat types. The z-value and the upper limit of the 95% confidence interval for the z-value were 4.64 and 5.32 degrees C, respectively. From the lowest temperature to the highest, the predicted D-values and the upper limits of their 95% prediction intervals (conservative D-values) for 57 to 61 degrees C were 241.2 and 321.1 s, 146.8 and 195.4 s, 89.3 and 118.9 s, 54.4 and 72.4 s, and 33.1 and 44.0 s. D-values and conservative D-values predicted for higher temperatures were 0.28 and 0.50 s for 70 degrees C and 0.041 and 0.073 s for 73.9 degrees C. Calculations with the conservative D-values predicted that cooking chicken meat according to current U.S. Department of Agriculture Food Safety and Inspection Service time-temperature guidelines will inactivate Korea/03 in a heavily contaminated meat sample, such as those tested in this study, with a large margin of safety.     

Inactivation of hepatitis A virus and a calicivirus by high hydrostatic pressure

Potential application of high hydrostatic pressure processing (HPP) as a method for virus inactivation was evaluated. A 7-log10 PFU/ml hepatitis A virus (HAV) stock, in tissue culture medium, was reduced to nondetectable levels after exposure to more than 450 MPa of pressure for 5 min. Titers of HAV were reduced in a time- and pressure-dependent manner between 300 and 450 MPa. In contrast, poliovirus titer was unaffected by a 5-min treatment at 600 MPa. Dilution of HAV in seawater increased the pressure resistance of HAV, suggesting a protective effect of salts on virus inactivation. RNase protection experiments indicated that viral capsids may remain intact during pressure treatment, suggesting that inactivation was due to subtle alterations of viral capsid proteins. A 7-log10 tissue culture infectious dose for 50% of the cultures per ml of feline calicivirus, a Norwalk virus surrogate, was completely inactivated after 5-min treatments with 275 MPa or more. These data show that HAV and a Norwalk virus surrogate can be inactivated by HPP and suggest that HPP may be capable of rendering potentially contaminated raw shellfish free of infectious viruses.     

Thermal inactivation of avian influenza virus and Newcastle disease virus in a fat-free egg product

High-pathogenicity avian influenza (HPAI) virus, low-pathogenicity avian influenza (LPAI) virus, virulent Newcastle disease virus (vNDV) and low-virulent Newcastle disease virus (lNDV) can be present on the eggshell surface, and HPAI viruses and vNDV can be present in the internal contents of chicken eggs laid by infected hens. With the increase in global trade, egg products could present potential biosecurity problems and affect international trade in liquid and dried egg products. Therefore, the generation of survival curves to determine decimal reduction times (D(T)-values) and change in heat resistance of the viruses (z(D)-value) within fat-free egg product could provide valuable information for development of risk reduction strategies. Thermal inactivation studies using A/chicken/Pennsylvania/1370/83 (H5N2) HPAI virus resulted in D(55)-, D(56)-, D(56.7)-, D(57)-, D(58)-, and D(59)-values of 18.6, 8.5, 3.6, 2.5, 0.4, and 0.4 min, respectively. The z(D)-value was 4.4 °C. LPAI virus A/chicken/New York/13142/94 (H7N2) had D(55)-, D(56.7)-, D(57)-, D(58)-, D(59)-, and D(60)-values of 2.9, 1.4, 0.8, 0.7, 0.7, and 0.5 min, respectively, and a z-value of 0.4 °C. vNDV avian paramyxoviruses of serotype 1 (AMPV-1)/chicken/California/212676/2002 had D(55)-, D(56)-, D(56.7)-, D(57)-, D(58)-, and D(59)-values of 12.4, 9.3, 6.2, 5, 3.7, and 1.7 min, respectively. The z(D)-value was 4.7 °C. lNDV AMPV-1/chicken/United States/B1/1948 had D(55)-, D(57)-, D(58)-, D(59)-, D(61)-, and D(63)-values of 5.3, 2.2, 1.1, 0.55, 0.19, and 0.17 min, respectively, and a z(D)-value of 1.0 °C. Use of these data in developing egg pasteurization standards for AI and NDV-infected countries should allow safer trade in liquid egg products.     

High-pressure processing applied to cooked sausages: bacterial populations during chilled storage

Vacuum-packaged cooked sausages were pressurized at 500 MPa for 5 or 15 min at mild temperature (65 degrees C) and later stored at 2 and 8 degrees C for 18 weeks. Counts of aerobic mesophiles and psychrotrophs, lactic acid bacteria, enterobacteria, Baird-Parker microbiota, and Listeria spp. were determined 1 day and 3, 6, 9, 12, 15, and 18 weeks after treatment and compared with those of cooked sausages treated at 80 to 85 degrees C for 40 min. Pressurization generated reductions of about 4 log CFU/g in psychrotrophs and lactic acid bacteria. Enterobacteria and Listeria proved the most pressure sensitive; insignificant or no growth was detected throughout the study. Heat treatment inactivated psychrotrophs and enterobacteria similarly to pressure treatment. Listeria monocytogenes and enterotoxigenic Staphylococcus aureus were not found in treated samples. In general, there was no significant difference in counts of any bacterial populations either among treatments or between storage temperatures. High-pressure processing at mild temperature is an effective preservation method that can replace heat pasteurization applied to some cooked meat and poultry products after packaging.     

Inactivation of hepatitis A virus by high-pressure processing: the role of temperature and pressure oscillation

Inactivation of hepatitis A virus (HAV) in Dulbecco's modified Eagle medium with 10% fetal bovine serum was studied at pressures of 300, 350, and 400 MPa and initial sample temperatures of -10, 0, 5, 10, 20, 30, 40, and 50 degrees C. Sample temperature during pressure application strongly influenced the efficiency of HAV inactivation. Elevated temperature (> 30 degrees C) enhanced pressure inactivation of HAV, while lower temperatures resulted in less inactivation. For example, 1-min treatments of 400 MPa at -10, 20, and 50 degrees C reduced titers of HAV by 1.0, 2.5, and 4.7 log PFU/ml, respectively. Pressure inactivation curves of HAV were obtained at 400 MPa and three temperatures (-10, 20, and 50 degrees C). With increasing treatment time, all three temperatures showed a rapid initial drop in virus titer with a diminishing inactivation rate (or tailing effect). Analysis of inactivation data indicated that the Weibull model more adequately fitted the inactivation curves than the linear model. Oscillatory high-pressure processing for 2, 4, 6, and 8 cycles at 400 MPa and temperatures of 20 and 50 degrees C did not considerably enhance pressure inactivation of HAV as compared with continuous high-pressure application. These results indicate that HAV exhibits, unlike other viruses examined to date, a reduced sensitivity to high pressure observed at cooler treatment temperatures. This work suggested that slightly elevated temperatures are advantageous for pressure inactivation of HAV within foods.     

Inactivation of Bacillus cereus spores in milk by mild pressure and heat treatments

The objective of this work was to study the germination and subsequent inactivation of Bacillus cereus spores in milk by mild hydrostatic pressure treatment. In an introductory experiment with strain LMG6910 treated at 40 degrees C for 30 min at 0, 100, 300 and 600 MPa, germination levels were 1.5 to 3 logs higher in milk than in 100 mM potassium phosphate buffer (pH 6.7). The effects of pressure and germination-inducing components present in the milk on spore germination were synergistic. More detailed experiments were conducted in milk at a range of pressures between 100 and 600 MPa at temperatures between 30 and 60 degrees C to identify treatments that allow a 6 log inactivation of B. cereus spores. The mildest treatment resulting in a 6 log germination was 30 min at 200 MPa/40 degrees C. Lower treatment pressures or temperatures resulted in considerably less germination, and higher pressures and temperatures further increased germination, but a small fraction of spores always remained ungerminated. Further, not all germinated spores were inactivated by the pressure treatment, even under the most severe conditions (600 MPa/60 degrees C). Two possible approaches to achieve a 6 log spore inactivation were identified, and validated in three additional B. cereus strains. The first is a single step treatment at 500 MPa/60 degrees C for 30 min, the second is a two-step treatment consisting of pressure treatment for 30 min at 200 MPa/45 degrees C to induce spore germination, followed by mild heat treatment at 60 degrees C for 10 min to kill the germinated spores. Reduction of the pressurization time to 15 min still allows a 5 log inactivation. These results illustrate the potential of high-pressure treatment to inactivate bacterial spores in minimally processed foods.     

畜禽鲜肉保鲜技术研究进展

畜禽原料肉是消费者常用的主要食物来源,传统畜禽肉以屠宰后直接销售的热鲜肉和在－18 ℃以下贮存的冷冻肉为主,随着禽流感、猪瘟等动物性流感的爆发和人们对食品质量安全要求的提高,畜禽肉的销售逐渐向冷藏生鲜肉形式转变。本文根据畜禽宰后肉品质变化机理和品质评价指标,综述了当前畜禽鲜肉的主要保鲜技术和发展趋势,旨在为其技术开展进一步研究和应用提供理论参考,推动畜禽养殖业和加工业的发展。     

Survival and growth of Yersinia enterocolitica strains inoculated in skimmed milk treated with high hydrostatic pressure

Four human pathogenic strains of Yersinia enterocolitica (serotypes O:1, O:3, O:8, and O:9) were inoculated (7-8 log CFU/ml) in UHT skimmed milk and treated at 300, 400, and 500 MPa for 10 min at 20 degrees C, and then kept at 8 degrees C to assess their evolution for 15 days. Treatments at 400 and 500 MPa caused the highest lethality, generally reaching counts below detection level (1 CFU/ml) in the culture media. At 300 MPa, the most baroresistant serotypes were O:3 and O:8. After 15 days of storage at 8 degrees C, Y. enterocolitica showed growth over 8 log (CFU/ml) in all treatments. Kinetic study of microbial inactivation in skimmed milk was performed with serotype O:8 at 300 MPa, showing a tailing after 35 min of pressure treatment.     

High pressure-induced inactivation of Qbeta coliphage and c2 phage in oysters and in culture media

High pressure (HP) treatment inactivates bacteria in shellfish, but its effects on viruses in shellfish have not yet been determined, although viral illness is frequently associated with shellfish consumption. The aim of this study was to investigate the baroresistance of two bacteriophage viruses, Qbeta coliphage and c2 phage, in oysters and in culture media. High numbers (>or=10(7) ml(-1) or g(-1)) of both phages were obtained in culture media and in oysters. Samples were HP treated at 200-800 MPa at 20 degrees C for up to 30 min. Little or no inactivation of either phage was observed in oysters or in culture media after treatment at 相似文献   

Resistance of poliovirus to inactivation by high hydrostatic pressures

Suspensions of poliovirus, a typical enteric virus, were subjected to high hydrostatic pressures of 200 MPa for 15 min, 400 MPa for 15 min, 600, MPa for 15 min, and 600 MPa for 1 h. Suspensions of respiratory adenovirus (a dissimilar virus type) were treated likewise for comparison. Whereas adenoviruses were inactivated at pressures of 400 MPa and above, poliovirus showed no statistically significant inactivation by hydrostatic pressures up to 600 MPa for 1 h. This finding may have implications for the use of high pressure in food processing, as poliovirus has physical similarities to significant foodborne pathogenic enteric viral agents such as hepatitis A virus.