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Multiple hurdles to commercial translation of next generation active food packaging technologies have been responsible for few successes among a sea of potentially promising innovations over the past 30 years. The development process involves numerous critical points originating at the basic research level, continuing through the raw material, package, and food manufacturing supply chains, and ending with consumer retail acceptance and purchase, which may fail to advance active packages into commercial markets. Complex regulatory and environmental sustainability concerns present a distinct global challenge to new technology transfer that will continue to directly impact economic growth and limit growth potential of next generation active food technologies. Understanding the identified challenges of active packaging technology commercial translation within the defined areas of academic research, manufacturing, regulatory (safety and environmental), and consumer acceptance will lead to systems based solutions for increasing availability of these technologies within a global marketplace.

Mycoplasma cynos is recognized as an emerging causative pathogen of canine infectious respiratory disease (CIRD) worldwide. We developed a new open-source real-time PCR (rtPCR) assay for M. cynos that performs well under standard rtPCR conditions. Primers and probes were designed to target the M. cynos tuf gene. Reaction efficiencies for the M. cynos tuf gene assay on 2 platforms were based on amplification of standard curves spanning 8 orders of magnitude: ABI 7500 platform, 94.3–97.9% ( r2 ≥ 0.9935); QuantStudio OpenArray platform, 119.1–122.5% ( r2 = 0.9784). The assay performed very well over a range of template input, from 109 copies to the lower limit of quantification at 4 copies of the M. cynos genome on the ABI 7500 platform. Diagnostic performance was estimated by comparison with an in-house legacy assay on clinical specimens as well as testing isolates that were characterized previously by intergenic spacer region (ISR) sequencing. Exclusivity was established by testing 12 other Mycoplasma species. To substantiate the high specificity of the M. cynos tuf gene assay, sequence confirmation was performed on ISR PCR amplicons obtained from clinical specimens. One ISR amplicon sequence revealed M. mucosicanis rather than M. cynos. The complete protocol of the newly developed M. cynos tuf assay is provided to facilitate assay harmonization.

Food wasted due to food spoilage remains a global challenge to the environmental sustainability and security of food supply. In food manufacturing, post-processing contamination of food can occur due to persistent bacterial biofilms, which can be resistant to conventional cleaning and sanitization. The objective was to characterize the efficacy of a polymeric coating in reducing

Changes in distribution patterns and demand for increased food quality have resulted in a desire to improve the shelf life of nonsterile dairy products. Refrigerated shelf life extension typically requires, at a minimum, reductions in the growth rate of spoilage microorganisms and subsequent product deterioration. Reducing initial bacterial loads, increasing pasteurization regimes, and reducing postprocessing contamination have all been employed with measured success. The use of antimicrobial additives has been discouraged primarily due to labeling requirements and perceived toxicity risks. Carbon dioxide (CO2) is a naturally occurring milk component and inhibitory toward select dairy spoilage microorganisms; however, the precise mechanism is not fully understood. CO2 addition through modified atmosphere packaging or direct injection as a cost-effective shelf life extension strategy is used commercially worldwide for some dairy products and is being considered for others as well. New CO2 technologies are being developed for improvements in the shelf life, quality, and yield of a diversity of dairy products, including raw and pasteurized milk, cheeses, cottage cheese, yogurt, and fermented dairy beverages. Here we present a comprehensive review of past and present research related to quality improvement of such dairy products using CO2.

Two-dimensional protein electrophoresis (2-DE) has undergone many technical improvements in the past 30 years, resulting in an analytical method that is unparalleled in the resolution of complex protein mixtures and capable of quantifying changes in protein expression from a wide variety of tissues and samples. The technique has been applied in many studies of neurologic disease to identify changes in spot patterns that correlate with disease. The true power of the technique emerges when it is coupled to state-of-the-art methods in mass spectrometry, which enable identification of the protein or proteins contained within a spot of interest on a 2-DE map. Investigators have successfully applied the technique to gain improved understanding of neurologic disease mechanisms in humans and in animal models and to discover biomarkers that are useful in the clinical setting. An important extension to these efforts that has not been realized thus far is the desire to profile changes in protein expression that result from therapy to help relate disease-modifying effects at the molecular level with clinical outcomes. Here we review the major advances in 2-DE methods and discuss specific examples of its application in the study of neurologic diseases.

The effect of pressurized (50 MPa) CO2 as a nonthermal process for bacterial reduction in raw skim milk was examined using a unique pressurized continuous flow system. The lethal effects of subcritical and super-critical CO2 applied at different temperatures and pressures toward total native psychrotrophic microbial populations, total inoculated Pseudomonas fluorescens, and total inoculated spore populations were studied and compared. Pressures between 10.3 and 48.3 MPa; temperatures of 15, 30, 35, and 40 degrees C; and CO2 concentrations of 0, 3, 66, and 132 g/kg of milk were studied. For both native populations and inoculated P. fluorescens, greater total microbial lethality was observed under supercritical CO2 conditions than under subcritical CO2 conditions. At 30 degrees C, there was no effect on total microbial lethality of increasing pressure up to 20.7 MPa with either 66 or 132 g/kg of CO2; at 35 degrees C, there was a positive relationship between pressure and lethality at CO2 levels of 132 g/kg, but no relationship at 66 g/kg of CO2. For total microbial populations and P. fluorescens, CO2 applied at 132 g/kg at 30 degrees C and pressures of 10.3 to 20.7 MPa resulted in an average standard plate count reduction of 3.81 and 2.93 log, respectively; at 35 degrees C and 20.7 MPa, maximum reductions achieved were 5.36 and 5.02 log, respectively. For both total microbial populations and inoculated P. fluorescens, CO2 exhibited a greater overall lethal effect at 132 g/kg than at 66 g/kg and a greater effect at 35 degrees C than at 30 degrees C. At 24.1 and 48.3 MPa and 40 degrees C, microbial lethality in raw aged milk treated with 3 g/kg of CO2 was not significantly different than that observed for uncarbonated milk; lethality achieved in milk treated with 132 g/kg of CO2 was significantly higher than that achieved in these 2 low-level CO2 treatments. No treatment studied had any significant impact on spore populations. Our work shows that, using the studied system, pressurized CO2 results in greater microbial lethality in milk above critical temperatures than below and suggests that a critical concentration threshold level of CO2 is required for lethal effects. Our work also suggests that supercritical CO2 processing in a continuous flow system can achieve reductions in some microbial populations equal to or better than that typically achieved during high-temperature, short-time pasteurization.

The effects of holding raw milk under carbon dioxide pressures of 68 to 689 kPa at temperatures of 5, 6.1, 10, and 20°C on the indigenous microbiota were investigated. These pressure-temperature combinations did not cause precipitation of proteins from the milk. Standard plate counts from treated milks demonstrated significantly lower growth rate compared with untreated controls at all temperatures, and in some cases, the treatment was microcidal. Raw milk treated with CO 2 and held at 6.1°C for 4 d exhibited reduced bacterial growth rates at pressures of 68, 172, 344, and 516 kPa; and at 689 kPa, demonstrated a significant loss of viability in standard plate count assays. The 689-kPa treatment also reduced gram-negative bacteria and total Lactobacillus spp. The time required for raw milk treated at 689 kPa and held at 4°C to reach 4.30 log 10 cfu/mL increased by 4 d compared with untreated controls. Total coliform counts in the treated milk were maintained at 1.95 log 10 cfu/mL by d 9 of treatment, whereas counts in the control significantly increased to 2.61 log 10 cfu/mL by d 4 and 2.89 log 10 cfu/mL by d 9. At d 8, Escherichia coli counts had not significantly changed in treated milk, but significantly increased in the control milk. Thermoduric bacteria counts after 8 d were 1.32 log 10 cfu/mL in treated milk and 1.98 log 10 cfu/mL in control milk. These data indicated that holding raw milk at low CO 2 pressure reduces bacterial growth rates without causing milk protein precipitation. Combining low CO 2 pressure and refrigeration would improve the microbiological quality and safety of raw milk and may be an effective strategy for shipping raw single strength or concentrated milk over long distances.

In total, 181 streptococci-like bacteria isolated from intramammary infections (IMI) were submitted by a veterinary clinic to Quality Milk Production Services (QMPS, Cornell University, Ithaca, NY). The isolates were characterized by sequence analysis, and 46 Lactococcus lactis ssp. lactis and 47 Lactococcus garvieae were tested for susceptibility to 17 antibiotics. No resistant strains were found for β-lactam antibiotics widely used in clinical practice (penicillin, ampicillin, and amoxicillin), and all minimum inhibitory concentrations (MIC) were far from the resistance breakpoints. Eight strains had MIC intermediate to cefazolin. The random amplification of polymorphic DNA (RAPD)-PCR fingerprint patterns showed a slightly higher heterogeneity for Lc. lactis ssp. lactis isolates than for Lc. garvieae isolates.

Lactococcus species are counted among a large and closely related group of environmental streptococci and streptococci-like bacteria that include bovine mastitis pathogenic Streptococcus, Enterococcus, and Aerococcus species. Phenotypic and biochemical identification methods can be inaccurate and unreliable for species within this group, particularly for Lactococcus spp. As a result, the incidence of Lactococcus spp. on the farm may have been historically underreported and consequently little is known about the clinical importance of this genus as a mastitis pathogen. We used molecular genetic identification methods to accurately differentiate 60 environmental streptococci and streptococci-like bacteria isolated from cows with high somatic cell count and chronic intramammary infection (IMI; >2 somatic cell scores above 4) among 5 geographically distinct farms in New York and Minnesota that exhibited an observed increase in IMI. These isolates were phenotypically identified as Streptococcus uberis and Streptococcus spp. Genetic methods identified 42 isolates (70%) as Lactococcus lactis ssp. lactis, including all 10 isolates originally phenotypically identified as Streptococcus uberis. Antibiotic inhibition testing of all Lc. lactis ssp. lactis showed that 7 isolates were resistant to tetracycline. In the present study, a predominance of Lc. lactis ssp. lactis was identified in association with chronic, clinical bovine IMI among all 5 farms and characterized antimicrobial resistance for treatment therapies. Routine use by mastitis testing labs of molecular identification methods for environmental streptococci and streptococci-like bacteria can further define the role and prevalence of Lc. lactis ssp. lactis in association with bovine IMI and may lead to more targeted therapies.

Background Streptococcus canis is an important opportunistic pathogen of dogs and cats that can also infect a wide range of additional mammals including cows where it can cause mastitis. It is also an emerging human pathogen. Results Here we provide characterization of the first genome sequence for this species, strain FSL S3-227 (milk isolate from a cow with an intra-mammary infection). A diverse array of putative virulence factors was encoded by the S. canis FSL S3-227 genome. Approximately 75% of these gene sequences were homologous to known Streptococcal virulence factors involved in invasion, evasion, and colonization. Present in the genome are multiple potentially mobile genetic elements (MGEs) [plasmid, phage, integrative conjugative element (ICE)] and comparison to other species provided convincing evidence for lateral gene transfer (LGT) between S. canis and two additional bovine mastitis causing pathogens (Streptococcus agalactiae, and Streptococcus dysgalactiae subsp. dysgalactiae), with this transfer possibly contributing to host adaptation. Population structure among isolates obtained from Europe and USA [bovine = 56, canine = 26, and feline = 1] was explored. Ribotyping of all isolates and multi locus sequence typing (MLST) of a subset of the isolates (n = 45) detected significant differentiation between bovine and canine isolates (Fisher exact test: P = 0.0000 [ribotypes], P = 0.0030 [sequence types]), suggesting possible host adaptation of some genotypes. Concurrently, the ancestral clonal complex (54% of isolates) occurred in many tissue types, all hosts, and all geographic locations suggesting the possibility of a wide and diverse niche. Conclusion This study provides evidence highlighting the importance of LGT in the evolution of the bacteria S. canis, specifically, its possible role in host adaptation and acquisition of virulence factors. Furthermore, recent LGT detected between S. canis and human bacteria (Streptococcus urinalis) is cause for concern, as it highlights the possibility for continued acquisition of human virulence factors for this emerging zoonotic pathogen.

Mammalian orthoreoviruses induce apoptosis in vivo and in vitro ; however, the specific mechanism by which apoptosis is induced is not fully understood. Recent studies have indicated that the reovirus outer capsid protein μ1 is the primary determinant of reovirus-induced apoptosis. Ectopically expressed μ1 induces apoptosis and localizes to intracellular membranes. Here we report that ectopic expression of μ1 activated both the extrinsic and intrinsic apoptotic pathways with activation of initiator caspases-8 and -9 and downstream effector caspase-3. Activation of both pathways was required for μ1-induced apoptosis, as specific inhibition of either caspase-8 or caspase-9 abolished downstream effector caspase-3 activation. Similar to reovirus infection, ectopic expression of μ1 caused release into the cytosol of cytochrome c and smac/DIABLO from the mitochondrial intermembrane space. Pancaspase inhibitors did not prevent cytochrome c release from cells expressing μ1, indicating that caspases were not required. Additionally, μ1- or reovirus-induced release of cytochrome c occurred efficiently in Bax −/− Bak −/− mouse embryonic fibroblasts (MEFs). Finally, we found that reovirus-induced apoptosis occurred in Bax −/− Bak −/− MEFs, indicating that reovirus-induced apoptosis occurs independently of the proapoptotic Bcl-2 family members Bax and Bak.

Inhibition of bacterial growth by dissolved carbon dioxide (CO2) has been well established in many foods including dairy foods. However, the effects of dissolved CO2 on specific growth parameters such as length of lag phase, time to maximum growth rate, and numbers of organisms at the stationary phase have not been quantified for organisms of concern in milk. The effect of dissolved CO2 concentrations of 0.6 to 61.4 mM on specific bacterial growth parameters in raw or single organism inoculated sterile milk was determined at 15 degrees C by conductance. Commingled raw or sterile milks were amended to a final concentration of 0.5 mg/ml each of urea and arginine HCl. Sterile milks were inoculated singly with one of six different microorganisms to a final concentration of approximately 10(2) to 10(3) cfu/ml; raw milk was adjusted to a final indigenous bacterial population of approximately 10(3) cfu/ml. Conductivity of the milk was recorded every 60 s over 4 to 5 d in a circulating apparatus at 15 degrees C. Conductivity values were fit to Gompertz equations and growth parameters calculated. Conductance correlated with plate counts and was satisfactory for monitoring microbial growth. Data fit the Gompertz equation with high correlation (R2 = 0.96 to 1.00). In all cases, dissolved CO2 significantly inhibited growth of raw milk bacteria, influencing lag, exponential, and stationary growth phases as well as all tested monocultures.

The effects of the addition of 11.9 mM CO2 on the growth of Bacillus cereus spores inoculated into sterile homogenized whole milk at 101 and 106 spores/ml and stored at 6.1 degrees C, was examined weekly for 35 d. Colony-forming units from CO2 treated inoculated milks decreased over 35 d at a rate similar to that of untreated inoculated milk, as defined by linear regression. Plate counts for treated and control milks inoculated at 10(1) cfu/ml were not significantly different on sampling d 0, 14, 21, and 28. Plate counts at d 7 were significantly different and counts at d 35 were at undetectable levels for both treated and control milks. Plate counts for milk inoculated at 10(6) cfu/ml were not significantly different on d 0, 28, and 35; they were significantly different on d 7, 14, and 21. There was no consistency as to whether the control or test milks were higher in counts on days when the differences were significant. Added CO2 reduced the pH of the milk from an average value of 6.61 to an average value of 6.31; however, this drop did not correlate with changes in any other parameter measured. These data suggest that moderate levels of CO2 do not enhance the outgrowth of B. cereus spores over long-term storage and do not increase the risk of foodborne illness due to the organism.

Les lipides des graines entieres de soja sont resistantes a l'oxydation a part certains lipides polaires. Les graines broyees et l'huile sont plus sensibles a l'oxydation

In view of their possible role in oxidative deterioration of seeds and pollen, organic free radicals were measured by electron spin resonance in embryonic axes and cotyledons of soybean [Glycine max (L.) Merr], embryo and endosperm fractions of corn [Zea mays L.] and pollen of cattail [Typha latifolia L.]. A pronounced decline in the radical signal ensued when hydration increased above about 7% (wet weight basis) in both the seed materials and in pollen. Moderate hydration of the soybean axis followed by drying led to a small decrease in organic free radicals compared to untreated material, especially if the desiccation step was performed under nitrogen. In a comparison of soybeans of various ages under normal storage, organic free radical levels in the axis showed little or no increase with age. In marked contrast, over 5 days of accelerated aging at 40°C and near-saturating humidity, organic radical levels approximately doubled in the axis. This pronounced increase in free radical content was not associated with a decrease in the proportion of polyunsaturated fatty acids. The data suggest that hydration of seed and pollen causes a release of free radicals from the trapped state.

The temperature limits for germination in Typha latifolia pollen lie within the range 4-40 degrees C. These limits correlate at the low-temperature end with the 'crystallization' of endogenous triacylglycerols and on the high-temperature end with the 'melting' of a gel-like lipid component in intact pollen. X-ray diffraction analysis was used to structurally characterize and to trace the latter gel-like lipid from the intact pollen through a range of pollen lipid fractions. We tentatively identify this component as a fatty acyl sterol ester and present evidence that it resides in the exine of the pollen grain. Its thermotropic behavior is insensitive to pollen hydration. The possibility of interpreting a crystalline lipid phase as being membrane-derived when in fact it originates from contaminating non-membranous neutral lipid is discussed. The total lipid content of T. latifolia pollen is 123 mg/g dry weight, of which 37% is polar lipid. The neutral lipid consists primarily of triacylglycerols and of the aforementioned sterol ester, which represents 0.34% (w/w) of pollen dry weight. The polar lipid fraction has phosphatidylcholine, phosphatidylethanolamine and phosphatidic acid as major components with lesser amounts of phosphatidylglycerol and phosphatidylinositol. Palmitic (16:0) and linoleic (18:2) acids, in a 1:2 molar ratio, constitute the major fatty acids of both polar and neutral lipid fractions with lesser amounts of linolenic (18:3), oleic (18:1) and stearic (18:0) acid in evidence.