Can the monitoring of animal welfare parameters predict pork meat quality variation through the supply chain (from farm to slaughter)?

The objective of this study was to assess the relationship between the animal welfare conditions evaluated through the supply chain and pork quality variation. A total of 4,680 pigs from 12 farms-5 animal welfare improved raising system (AWIRS) and 7 conventional raising system (CON) farms-were assessed from farm to slaughter through a comprehensive audit protocol merging the European Welfare Quality, the Canadian Animal Care Assessment, and American Meat Institute audit guide criteria. At the abattoir, a subsample of 1,440 pigs (120 pigs/farm) was randomly chosen out of 24 loads (2 farms per wk) transported by 2 drivers (driver A and driver B) for the assessment of stunning effectiveness, carcass bruises, blood lactate levels, and meat quality traits. Meat quality was assessed in the longissimus lumborum (LL) muscle 24 h postmortem by measuring ultimate pH (pHu), color (L*, a*, and b*), and drip loss. Data were analyzed by the MIXED, GLIMMIX, and NAPAR1WAY procedures of SAS. Spearman correlations were calculated to determine the relationship between audit scores and meat quality traits. Better animal welfare conditions, as showed by greater final scores for good housing (GHo; = 0.001) and good health ( = 0.006) principles, were recorded at AWIRS farms. Pigs from AWIRS farms handled by driver B displayed a greater percentage of turning back ( = 0.01) and slips ( < 0.001) during unloading and a greater ( = 0.02) frequency of falls in the stunning chute. A greater ( = 0.02) reluctance to move at loading was found in CON pigs loaded by driver A compared with driver B, whereas a greater ( < 0.001) reluctance to move was found in these pigs at unloading when they were unloaded by driver B. Drip loss was higher ( = 0.003) and pale, soft, and exudative pork percentage was greater ( < 0.001) in the LL muscle of the heavier AWIRS pigs. The GHO principle was best correlated with pHu ( = -0.75, = 0.01) and Minolta L* value ( = 0.87, < 0.001) of the LL muscle. Overall, drip loss variation in the LL muscle was correlated with the frequency of slips at unloading ( = 0.63, = 0.001) and in the restrainer area ( = 0.74, < 0.001). The results of this study showed that the quality of the raising system and truck driver skills as assessed by animal welfare audit protocols are important sources of variation in the behavioral response of pigs to preslaughter handling and may affect pork quality variation. However, the different live weight between CON and AWIRS pigs may have biased the meat quality results in this study.

Hand-held lactate analyzer as a tool for the real-time measurement of physical fatigue before slaughter and pork quality prediction.

The objectives of this study were to assess the relationship between blood lactate variation measured at the plant, and pork quality variation on a large sample size and under commercial preslaughter handling conditions. A total of 600 pigs were randomly chosen on arrival at a commercial slaughter plant and blood samples taken from the ear vein at unloading (UN), after lairage (LA), in the restrainer (RE; before stunning) and at exsanguination (EX) were analysed for lactate content using a Lactate Scout Analyzer (LSA). In order to have a large range of measures, pigs were distributed into two groups; one kept in lairage overnight (G1) and the other for 2 to 3 h (G2) before slaughter. Meat quality was assessed in the Longissimus thoracis (LT), Semimembranosus (SM) and Adductor (AD) muscles by measuring the pH 30 min postmortem (pH1) and at 24 h postmortem (pHu), the colour and the drip loss. Blood lactate levels did not differ between G1 and G2 (P>0.05). A reduced muscle lactate and glucose contents (P=0.02 and P=0.004, respectively) resulting in a lower (P<0.001) glycolytic potential (GP) was observed in the LT muscle of G1 pigs when compared with G2 loins. In the LT muscle of G1 pigs, the lower GP resulted in an increased pHu (r=-0.67; P<0.001), decreased drip loss (r=0.57; P<0.001) and darker colour (r=0.50; P<0.001) compared with G2. In both G1 and G2 pigs, the lower GP was correlated to higher pHu value in the SM and AD muscles (r=-0.73; P<0.001). The greatest correlation was observed in G2 between blood lactate levels at LA and pHu value of the SM and AD muscles (r=0.46 and r=0.44, respectively; P<0.001 for both muscles). The second greatest correlation was found between blood lactate levels at EX and pH1 value in the SM muscle in both groups (r=-0.37 and r=-0.41, respectively; P<0.001 for both groups). Based on the results of this study, it appears that blood lactate levels, as measured by the LSA, reliably reflect the physiological response of pigs to perimortem stress and may help explain the variation in pork quality.

Whole-genome transcriptional analysis of Escherichia coli during heat inactivation processes related to industrial cooking.

Escherichia coli K-12 was grown to the stationary phase, for maximum physiological resistance, in brain heart infusion (BHI) broth at 37°C. Cells were then heated at 58°C or 60°C to reach a process lethality value \[\mathbf{\left(}{{\mathit{F}}^{\mathit{o}}}_{\mathbf{70}}^{\mathbf{10}}\mathbf{\right)} \] of 2 or 3 or to a core temperature of 71°C (control industrial cooking temperature). Growth recovery and cell membrane integrity were evaluated immediately after heating, and a global transcription analysis was performed using gene expression microarrays. Only cells heated at 58°C with F(o) = 2 were still able to grow on liquid or solid BHI broth after heat treatment. However, their transcriptome did not differ from that of bacteria heated at 58°C with F(o) = 3 (P value for the false discovery rate [P-FDR] > 0.01), where no growth recovery was observed posttreatment. Genome-wide transcriptomic data obtained at 71°C were distinct from those of the other treatments without growth recovery. Quantification of heat shock gene expression by real-time PCR revealed that dnaK and groEL mRNA levels decreased significantly above 60°C to reach levels similar to those of control cells at 37°C (P < 0.0001). Furthermore, despite similar levels of cell inactivation measured by growth on BHI media after heating, 132 and 8 genes were differentially expressed at 71°C compared to 58°C and 60°C at F(o) = 3, respectively (P-FDR < 0.01). Among them, genes such as aroA, citE, glyS, oppB, and asd, whose expression was upregulated at 71°C, may be worth investigating as good biomarkers for accurately determining the efficiency of heat treatments, especially when cells are too injured to be enumerated using growth media.

Effects of trailer design on animal welfare parameters and carcass and meat quality of three Pietrain crosses being transported over a long distance.

This study aimed at evaluating the effects of trailer design on stress responses and meat quality traits of 3 different pig crosses: 50% Pietrain breeding with halothane (HAL)(Nn) (50Nn); 50% Pietrain breeding with HAL(NN) (50NN); and 25% Pietrain breeding with HAL(NN) genotype (25NN). Over a 6-wk period, pigs (120 pigs/crossbreed) were transported for 7 h in either a pot-belly (PB) or flat-deck (FD) trailer (10 pigs/crossbreed(-1)·trailer(-1)·wk(-1)). Temperature (T) and relative humidity (RH) were monitored in each trailer. Behaviors during loading and unloading, time to load and unload, and latency to rest in lairage were recorded, whereas a sub-population of pigs (4 pigs/crossbreed(-1)·trailer(-1)·wk(-1)) was equipped with gastro-intestinal tract (GIT) temperature monitors. Blood samples were collected at exsanguination for measurement of cortisol, creatine kinase (CK), lactate, haptoglobin, and Pig-MAP concentrations. Meat quality data were collected at 24 h postmortem from the LM and semimembranosus (SM) and adductor (AD) muscles of all 360 pigs. Greater T were recorded in the PB trailer during transportation (P = 0.006) and unloading (P < 0.001). Delta GIT temperature was greater (P = 0.01) in pigs unloaded from the PB. At loading, pigs tended to move backwards more (P = 0.06) when loaded on the FD than the PB trailer. At unloading, an interaction was found between trailer type and crossbreed type, with a greater (P < 0.01) frequency of overlaps in 50NN and 25NN pigs and slips/falls in 50Nn and 50NN pigs from the FD than the PB trailer. Cortisol concentrations at slaughter were greater (P = 0.02) in pigs transported in the PB than FD trailer. Greater lactate concentrations were found in 50Nn and 50NN pigs (P = 0.003) and greater CK concentrations (P < 0.001) in 50Nn pigs. As expected, 50Nn pigs produced leaner (P < 0.001) carcasses, with greater (P = 0.01) dressing percentages, as well as lower (P < 0.001) ultimate pH values and greater (P < 0.001) drip loss percentages in the LM and greater (P = 0.002) drip losses and a paler color (greater L* values, P = 0.02) in the SM than 50NN pigs. When used for long distance transportation under controlled conditions, the PB trailer produced no detrimental effects on animal welfare or pork quality. Pigs with 50% Pietrain crossbreeding appear to be more responsive to transport stress, having the potential to produce acceptable carcass and pork quality, provided pigs are free of the HAL gene.

Mycobacterium avium subsp. paratuberculosis in dairy products, meat, and drinking water.

Mycobacterium avium subsp. paratuberculosis (Map) is the cause of Johne's disease, a chronic infection of the gut, in ruminant animals that provide milk and/or meat for human consumption. Map also may be involved in Crohn's disease and type 1 diabetes in humans. Although the role of Map in human diseases has not been established, minimizing the exposure of humans to the organism is considered desirable as a precautionary measure. Infected animals can shed Map in feces and milk, and the organism can become disseminated in tissues remote from the gut and its associated lymph nodes. The presence of at least some Map in raw milk and meat and in natural waters is likely, but the numbers of Map in those foods and waters should be reduced through cooking or purification. The available information relating to Map in milk and dairy products, meats, and drinking water is reviewed here for assessment of the risks of exposure to Map from consumption of such foods and water.

Shelf life of pork from five different quality classes.

A total of 117 loins were selected on the cutting line at 24h post-mortem to study the long term shelf life (35 days, 4 degrees C) of vacuum packaged pork from five different quality classes (PSE: pale, soft, exudative; PFN: pale, firm, non-exudative; RSE: red, soft, exudative; RFN: red, firm, non-exudative; and DFD: dark, firm, dry). The microbial load at 0 d was not significantly different (P>0.05) among the pork quality classes, indicating that the initial microflora was influenced by the dressing conditions at the plant, not by the meat quality class. But after 35 d of storage, total aerobic mesophilic and presumptive lactic acid bacteria counts were higher (P<0.05) in DFD pork due to its higher ultimate pH. RSE was the second quality class most susceptible to spoilage, whereas PFN, RFN and PSE pork had similar microbial loads. Further research is needed to elucidate the causes of the shorter shelf life in RSE pork.

Prevalence on beef carcasses of Mycobacterium avium subsp. paratuberculosis DNA.

Fifty samples were collected from each of skinned and dressed carcasses, from each of culled beef breeding cows and fed beef cattle <18 months old at two beef packing plants A and B, and from culled dairy cows at a packing plant C. The 450 samples were collected by swabbing an area of about 1000 cm2 in the anal region of each carcass. DNA extracted from each swab was tested for the IS900 and F57 sequences of the Mycobacterium avium subsp. paratuberculosis (MAP) genome by two stage, nested polymerase chain reaction (PCR) procedures. An internal amplification control (IAC) was detected in 45 or more of each group of 50 DNA preparations. IS900 and F57 were detected in some IAC-positive preparations from all and all but one of the groups of carcasses, respectively. Of the IAC-positive preparations in each group, between 6 and 54% were positive for IS900, and between 4 and 20% were positive for F57. When preparations were tested by single stage, quantitative PCR procedures, IS900 was detected in two samples but F57 was detected in none. The MAP DNA on carcasses was probably derived from small numbers of MAP from the environment that contaminated the animals' hides.

Shiga toxin production by sausage-borne Escherichia coli O157:H7 in response to a post-processing in vitro digestion challenge.

In order to study the effects of the fermentation-drying procedure and subsequent in vitro digestion on Shiga toxins (Stx) production by Escherichia coli O157:H7, dry sausages were inoculated during the formulation step with pure cultures of strains 5-1 and ATCC 43895. The inoculated sausages were submitted to a minimum (30 min, pH between 3.1 and 3.5) or a maximum (120 min at stepwise adjusting the pH downward) gastric challenge followed by a 240-min pancreatic challenge at pH 8.0 and 37 degrees C. Production of toxins by the overnight culture controls, assessed using the Vero cell assay, was dependent on the pathogen cell concentration. The effect of cell concentration was not relevant in sausage samples and data showed: (a) higher Stx production in contaminated sausage samples than in overnight cultures; (b) the lowest Stx levels were detected with undigested sausage samples; (c) the maximum gastric challenge enhanced Stx production, compared to minimally digested and undigested samples. Reverse transcriptase polymerase chain reaction (RT-PCR) performed on extracts from inoculated, digested (4.5-6 h process) and undigested sausages produced amplicons for both stx1 and stx2 mRNA, suggesting that post-stress expression of stx genes had occurred. Our data suggest that sub-lethal stresses imposed by the fermentation-drying procedure and subsequent digestion of ingested food (i.e. contaminated sausages) may affect the degree to which the surviving E. coli O157:H7 cells express their virulence in vivo.

Effect of feed texture, meal frequency and pre-slaughter fasting on carcass and meat quality, and urinary cortisol in pigs.

Carcass and meat quality traits, and urinary cortisol variation was studied in 96 barrows assigned to the following treatments: feed texture (FT; mash vs. pellets), meal frequency (MF; 2 vs. 5 meals per day) and fasting time (F; 4, 14 and 24h) according to a 2×2×3 factorial design. Pigs fed mash, receiving feed five times a day and fasted for 24h before slaughter had lower carcass dressing yield (P<0.001). A higher (P<0.05) bruise score was found on carcasses from pigs fasted for 14 and 24h and fed either pelleted or mashed feed five times per day. The pH(u) value in the Longissimus muscle increased (P<0.05) with increasing fasting time, whereas in the Adductor muscle it was higher (P<0.05) in pigs fed with pellets in two meals per day and fasted for 24h. Urinary cortisol tended to be higher in pigs fasted for 14h compared to those fasted for 4 (P=0.10) and 24h (P=0.06). The results of this study show a significant influence of pellet feeding on carcass yield in fasted pigs, while the effects of pre-slaughter fasting time on meat quality traits were limited.

Combined effect of ascorbic acid and gamma irradiation on microbial and sensorial characteristics of beef patties during refrigerated storage.

The present study was undertaken to evaluate the effect of ascorbic acid concentrations (0.03 to 0.5%) and irradiation doses (0.5 to 4 kGy) on microbial growth, color coordinates (L, a, and b), and sensory characteristics (taste and odor) of beef patties during storage at 4 +/- 1 degrees C. Ascorbic acid was also compared to citric acid at a similar pH value in order to differentiate the effects of ascorbic acid from those of pH reduction. Results showed significant reduction (p< or = 0.05) of aerobic plate counts (APCs) and total coliforms, and a significant interaction (p< or = 0.05) between ascorbic acid and irradiation dose was observed. The irradiation treatment had detrimental effects on redness, yellowness, and hue angle values of meat. However, incorporation of ascorbic acid into the meat before irradiation resulted in significant (p< or = 0.05) stabilization of color parameters. The color improvement obtained with ascorbic acid was not related to the pH reduction. Also, no significant detrimental effect on taste or odor was found in irradiated samples containing ascorbic acid.

Description of a simple detection assay for in situ production of bacteriocin on meat.

Using a modification of the agar diffusion assay, in situ bacteriocin production on meat was analyzed using cooked meat medium (CMM) and sterile pork tissue (lean and fat) with Carnobacterium piscicola LV17 as the producer and Carnobacterium divergens LV13 as the indicator strains. Contrary to what is observed in APT broth, bacteriocin production by C. piscicola LV17 occurred with growth at low inoculum levels (< or =10(4) CFU/cm2 or g of meat) on disks (10 cm2) of pork fat tissue (pH 6.58) and on CMM particles (pH 7.0) but not on disks of lean tissue (pH 5.61). The assays described in this study do not required sophisticated equipment and would be useful to study bacteriocin production on meat products stored under various conditions.

Adaptation of Lactobacillus alimentarius to environmental stresses.

Lactobacillus alimentarius BJ33 has been tested for its biopreservative capacities to improve quality and safety in many meat products. The combination of different preservatives such as NaCl, glucono-delta-lactone and citric acid with this protective culture during the manufacture of sausages represent an interesting alternative to control microbial spoilage and to extend product shelf life. The use of these preservatives may also limit the growth of L. alimentarius. In this study, the sublethal doses of these preservatives were determined and tested in combination to verify if the organism was able to adapt to these stresses. The sublethal doses of gluconic acid, citric acid, and NaCl were 100-110 mM, 50-55 mM and 8%, respectively. When the culture was first grown in MRS broth containing citric acid (50 or 55 mM) or gluconic acid (100 or 110 mM) and then transferred in MRS broth containing NaCl (8%), only limited growth was observed (O.D.(600 nm) = 0.2-0.3) after 6 days at 30 degrees C. However, when the culture was first grown in NaCl and then transferred in MRS broth containing gluconic or citric acid, growth was observed after 1 day (O.D.(600 nm) = 0.4-0.5) and after 5 days an O.D.(600 nm) of 0.8 was reached. Cell filamentation was also observed under electron microscopy when cells were grown for 2 days in presence of gluconic and citric acid at their sublethal doses and with a combination of 18 mM gluconic acid and 37 mM citric acid, but cellular elongation was not observed with cultures exposed to 8% NaCl. These results suggest that two different adaptation mechanisms are induced in L. alimenatrius when treated with organic acids and NaCl.

Shelf life of ground poultry meat stored under modified atmosphere.

The shelf life of ground chicken and turkey meat packaged under a modified atmosphere containing O2 and a high level of CO2 (62% CO2, 8% O2, and 30% N2; gas-1), or a gas mixture without O2 (20% CO2 and 80% N2; gas-2) was evaluated for 20 d at 1 C. Meat packaged under gas-2 maintained a higher a* value (redness) throughout the experiment and generally had a more appealing color than the meat packaged using gas-1. Microbial populations were assessed after 8, 12, and 15 d of storage. Total aerobic mesophilic counts were higher in chicken meat than in turkey throughout storage. Coliforms and Escherichia coli counts were lower in meat packaged under gas-1. After 15 d of storage at 1 C, Brochothrix thermosphacta was detected only in ground chicken meat packaged using gas-2. Meat packaged under the gas mixtures tested had similar counts for presumptive pseudomonads, Staphylococcus aureus, and lactic acid bacteria. These results indicate that an appropriate gas mixture can maintain a desirable color in ground poultry meat but offers no guarantees with respect to the microbial profile of meat.

Transcriptional analysis and regulation of carnobacteriocin production in Carnobacterium piscicola LV17.

Bacteriocin production by Carnobacterium piscicola LV17 (carnobacteriocin, Cbn) depends on the level of inoculation when grown in liquid medium. With an inoculum of > or = 10(6) colony-forming units per ml (cfu/ml), bacteriocin production is observed during exponential growth, whereas with < or = 10(4) cfu/ml no bacteriocin is detected even when the culture has reached stationary phase. Using pure bacteriocins, it was demonstrated that bacteriocin production is autoregulated. To understand how bacteriocin production is regulated at the molecular level, cell-free supernatant from a bacteriocin-producing culture was added to fresh medium at 1% (v/v) together with a non-producing inoculum (10(4) cfu/ml), to induce bacteriocin production (induced culture). Northern analysis revealed major transcripts of 0.35, 1.5 and 1 kb for carnobacteriocins A, B2 and BM1, respectively, indicating that regulation of bacteriocin production by inoculum size occurs at the transcriptional level. Primer extension demonstrated that transcription initiated from the same promoters with the induced culture as with the positive control (culture inoculated at 10(7) cfu/ml). Quantitative phosphorimager analysis of the primer extension products indicated that cbnA transcript was more abundant than cbnB2 or cbnBM1.

Effect of Feeding Lactic Acid Bacteria and Fermented Milk on Specific and Nonspecific Immune Reponses of Mice Infected With Klebsiella pneumoniae AD-1.

The diets of six groups of weaned mice were supplemented with ultra high temperature (UHT) milk containing a washed suspension of lactic acid bacteria (mixture of 8 strains) or with UHT milk fermented by the same strains and heat-treated or not. Control groups received physiological saline or UHT milk only. The mice were infected intranasally by Klebsiella pneumoniae AD-1 on the 13th d of feeding. The effect on the immune system (specific and nonspecific) before and after infection was evaluated by measuring the phagocytosis of alveolar macrophages (using zymosan particles) and by measuring of total immunoglobulin G and A levels in serum and in pulmonary fluid (using the enzyme-linked immunosorbent assay method). Postinfection survival was 0.7 d longer for mice receiving fermented milk than for the saline control group. The percent phagocytosis did not vary significantly, while serum immunoglobulin G levels differed between mice fed fermented milk and those fed bacterial suspensions in unfermented milk. Fermentation appears to be essential for the beneficial effects on the immune system and survival time; this effect no longer occurs after pasteurization of fermented milk.