B. W. Sheldon

Efficacy of nisin-coated polymer films to inactivate Salmonella Typhimurium on fresh broiler skin.

Nisin is an antimicrobial peptide produced by the food-grade microorganism Lactococcus lactis subsp. lactis. This peptide inhibits many gram-positive bacteria, and when combined with chelating agents it inhibits gram-negative bacteria such as Salmonella sp. The efficacy of packaging films treated with nisin-containing formulations to reduce Salmonella contamination of fresh broiler drumstick skin and increase the refrigerated shelf life was investigated. Three films (5.1 cm2) of varying hydrophobicities (polyvinyl chloride [PVC], linear low density polyethylene, nylon) were coated with one of three liquid formulations (pH 3.5 to 3.8) composed of 100 microg/ml nisin and varying concentrations of citric acid, EDTA, and Tween 80. The treated films were applied either wet or dry to 5.1-cm2 broiler drumstick skin samples inoculated with a nalidixic acid-resistant (NAr) strain of Salmonella Typhimurium. After incubation at 4 degrees C for 24 h the populations of surviving Salmonella TyphimuriumNAr organisms were recovered from the skin and film samples using a rinse procedure and enumerated on brain heart infusion agar containing 800 ppm NA. Log reductions (untreated versus treated skin) in Salmonella TyphimuriumNAr populations ranged from 0.4 to 2.1. Treatment formulation compositions and wet versus dry treatment application also influenced the extent of kill. The shelf life of refrigerated broiler drumsticks was extended by 0.6 to 2.2 days following a 3-min immersion in a nisin-containing treatment solution and subsequent storage in a foam tray pack containing a nisin-treated PVC overwrap and a nisin-treated absorbent tray pad. These findings demonstrated that Salmonella Typhimurium and spoilage microorganism populations on the surface of fresh broiler skin and drumsticks can be significantly reduced using immersion treatments, absorbent tray pads, and packaging films treated with nisin-containing formulations.

Inhibition of Salmonella on poultry skin using protein- and polysaccharide-based films containing a nisin formulation

The objective of this study was to examine the use of protein- arid polysaccharide-based films containing bacteriocin formulations for inhibiting salmonellae on fresh broiler skin. The lethality of the films containing a nisin-based formulation was determined against Salmonella Typhimurium-contaminated broiler drumstick skin samples coated with the film. In the first study, varying concentrations of nisin (0, 100, 300, and 500 microg/ml) plus 3% citric acid, 5.0 mM EDTA, and 0.5% Tween 80 were incorporated into 0.5% calcium alginate films at a 20% level (wt/wt) and then applied to Salmonella TyphimuriumNAr-contaminated skin samples (log10 5.0) at a 1:2 weight ratio (film:skin). Salmonella TyphimuriumNAr skin population reductions ranged from 1.98 to 3.01 log cycles after a 72-h exposure at 4 degrees C. In comparison to the 0- and 100-microg/ml nisin concentrations, significantly greater population reductions were achieved at nisin concentrations of 300 and 500 microg/ml. In related studies, the 500-degreesg/ml nisin formulation was incorporated into 0.75 and 1.25% agar gels and applied to contaminated broiler drumstick skin samples (log10 7.0). Salmonella TyphimuriumNAr skin population reductions following a 96-h exposure at 4 degrees C were 1.8-(1.25% agar gel) and 4.6-log cycles (0.75% agar gel). These results demonstrated that the inclusion of nisin-based treatments into either calcium alginate or agar gels that were subsequently applied to contaminated broiler drumstick skin yielded significant Salmonella TyphimuriumNAr population reductions ranging between 1.8 to 4.6 log cycles after 72 to 96 h of exposure at 4 degrees C. The level of kill was affected by film type and gel concentration (i.e., gel network formation), exposure time, and nisin concentration.

Antioxidant properties of individual phospholipids in a salmon oil model system

The antioxidant properties of phospholipids (PL) in a refined salmon oil model system were measured by determining changes in the 2-thiobarbituric acid number and decreases in the ratio of docosahexaenoic acid (DHA)/palmitic acid (22:6/16:0) of a fish oil system incubated at 180°C for up to 3 h. The more phosphatidylcholine (PC) added to the oil system, the higher the oxidative stability obtained. The order of effectiveness of commercial phospholipids in inhibiting oxidation and the loss of polyunsaturated fatty acids was as follows: sphingomyelin (SPH)=lysophosphatidylcholine (LPC)=phosphatidylcholine (PC)=phosphatidylethanolamine (PE)>phosphatidylserine (PS)>phosphatidylinositol (PI)>phosphatidylglycerol (PG)>control salmon oil. Nitrogen containing PL, including PE, PC, LPC and SPH, were equally effective in exerting greater antioxidant properties than PS, PG and PI. The inverse relationship observed between the oxidation index (C22:6/C16:0) and color intensity for treatments following 2 h of heating suggests that Maillard-type reaction products may have contributed to the oxidative stability of PL-supplemented fish oils.

A rapid method for determining the oxidation of n-3 fatty acids

The stability of unsaturated fatty acids to oxidation was monitored by following gas chromatographic (GC) analyses of headspace volatiles in comparison to changes in polyunsaturated fatty acids (PUFA) and increases in malonaldehydevia the 2-thiobarbituric (TBA) assay. Pure standards of linoleic acid (Lo) and n-3 fatty acids [eicosapentaenoic (EPA) and docosahexaenoic acid (DHA)] were added to headspace vials, equilibrated in air for 10 min, followed by heating at 80°C in teflon-capped vials for different time intervals. Headspace analysis showed increases in acetaldehyde, propenal, and propanal, corresponding to the oxidation of n-3 fatty acids, whereas hexanal production corresponded to losses of linoleic acid. The analysis of propanal by GC-headspace after only five minutes of heating appeared to be the most effective method of monitoring the oxidation of n-3 fatty acids, as indicated by correlations between TBA values and loss of PUFA. The oxidation of Lo, EPA and DHA appeared to be a function of the number of double bonds. Correlations between PUFA depletion, TBA values and volatile formation indicate that under the prescribed conditions of this experiment, GC-headspace analysis of propanal and pentane/hexanal is an excellent method for following the oxidation of selected n-3 fatty acids and linoleic acid.

Immersion heat treatments for inactivation of Salmonella enteritidis with intact eggs

The effects of water‐bath immersion heat treatments on the inactivation of Salmonellaenteritidis within intact shell eggs were evaluated. Six pooled strains of Salm. enteritidis (ca 3×108 cfu, inoculated near the centre of the yolk) were completelyinactivated within 50–57·5 min at a bath temperature of 58°C and within 65–75min at 57°C (an 8·4 to 8·5‐D process per egg). Following the initial 24 to35‐min come‐up period, semilogarithmic survivor curves obtained at 58 and 57°C yieldedapparent decimal reduction times (D‐values) of 4·5 and 6·0 min, respectively.Haugh unit values increased during heating, while yolk index and albumen pH values wereunaffected. Albumen clarity and functionality were affected by the thermal treatments; therefore,extended whip times would be required for meringue preparation using immersion‐heated eggwhites. Immersion‐heated shell eggs could provide Salmonella‐free ingredients for thepreparation of a variety of minimally‐cooked foods of interest to consumers and foodserviceoperators.

Effects of phospholipids on lipid oxidation of a salmon oil model system

Total lipid (TL), neutral lipid (NL), and phospholipid (PL) fractions were extracted from bluefish (Pomatomus saltatrix) white and dark muscle with skin. The effects of each fraction on the oxidative stability of a refined salmon oil model system was measured by monitoring changes in the 2-thiobarbituric acid assay and decreases in the ratio of docosahexaenoic acid (DHA) to palmitic acid (C22:6/C16:0) following incubation at 55°C or 180°C. Phospholipid fractions at 2.5% and 5.0% (wt/wt) of oil improved the oxidative stability of oils incubated at both temperatures compared to controls, TL- and NL-supplemented oils at similar concentrations. Phospholipid fractions exhibiting antioxidant properties contained an average of 34% DHA as compared to only 15% in the NL and TL fractions.

Influence of Housing System, Grain Type, and Particle Size on Salmonella Colonization and Shedding of Broilers Fed Triticale or Corn-Soybean Meal Diets

Salmonella colonization in poultry may be influenced by grain type and particle size. Broilers reared either in nonlitter cage-based housing or in a conventionally floored litter house from 0 to 42 d were assigned to 1 of 4 dietary treatments: 1) ground corn-soybean meal (C, 560 microm), 2) coarsely ground corn-soybean meal (CC, >1,700 microm), 3) ground triticale-soybean meal (T, 560 microm), or 4) whole triticale-soybean meal (WT). A 4-strain cocktail of Salmonella enterica was orally gavaged into each chick at placement. Growth performance, cecal and fecal Salmonella populations, gizzard and proventriculus pH, intestinal size, jejunum histomorphometry, and carcass yields were measured. Broilers responded differently to the dietary treatments according to the housing system used. At 42 d, birds reared on litter and fed ground grain had greater BW than those fed coarse grain (2.87 vs. 2.71 kg), whereas cage-reared broilers fed ground triticale were heavier than those fed corn (2.75 vs. 2.64 kg). Broilers raised on litter had a better feed conversion ratio than those raised in cages (1.71 vs. 1.81 g/g). Independent of the housing system, relative eviscerated carcass weights of birds fed T and C were heavier than those of CC- and WT-fed broilers (762 vs. 752 g/kg). Generally, the jejunum villus area and mucosal depth were larger, whereas the small intestine was lighter and shorter in broilers raised on litter. Relative gizzard weights of broilers raised on litter and fed the coarser diets were heavier than those of broilers reared in cages and fed finely ground diets. Feeding whole or coarsely ground grains decreased cecal Salmonella populations in 42-d-old broilers (3.8, 3.9, 4.4, and 4.4 log most probable number/g for CC, WT, C, and T, respectively). Additionally, 42-d-old broilers reared on litter had lower cecal Salmonella populations than those in cages (3.8 vs. 4.4 log most probable number/g). In conclusion, as a feed ingredient, triticale is a good alternative to corn, resulting in improved BW and reduced Salmonella colonization. Broilers raised on litter may have achieved lower cecal Salmonella populations than caged birds because access to litter may have modulated the intestinal microflora by increasing competitive exclusion microorganisms, which discouraged Salmonella colonization.

Efficacy of Optimized Nisin-Based Treatments to Inhibit Salmonella typhimurium and Extend Shelf Life of Broiler Carcasses†

Nisin is an antimicrobial peptide produced by Lactococcus lactis subsp. lactis . Nisin exhibits a broad spectrum of inhibitory activity against gram-positive microorganisms such as Listeria monocytogenes and Clostridium botulinum . In previous studies, a method was developed using nisin in combination with food-grade chelating agents to inactivate Salmonella species and other gram-negative bacteria. The objectives of this study were to determine the efficacy of several optimized nisin-containing preparations for reducing the population of Salmonella typhimurium NAR on broiler drumstick skin and whole drumsticks and extending the shelf life of broiler drumsticks. In previous studies, a simplex algorithm was used to optimize the biocidal activity of nisin towards Salmonella by adjusting formula pH and adding varying concentrations of chelating agents (EDTA, citric acid) and a surfactant (Tween 20). From these studies, four optimal treatments were identified. These were tested more extensively in the present study. Significant reductions in viable S. typhimurium NAR populations on broiler drumstick skin were achieved and ranged from 3.1 to 4.9 log10 cycles following immersion for 30 min at 25°C. The inhibitory activities of these four treatments against S. typhimurium NAR-contaminated drumsticks were also compared to treatments with 20 ppm chlorine. Numbers of survivors following a 30-min dip ranged from <10 to 2.57 × 101 organisms per ml of skin rinse for the nisin formulations versus 1.32 × 102 organisms per ml on the chlorine-treated drumsticks. In other studies, the shelf life of refrigerated broiler drumsticks was extended by 1.5 to 3 days following immersion for 30 min in one of the optimized nisin-containing treatments in comparison to drumsticks dipped in sterile distilled, deionized water.

Thermal Resistance of Salmonella spp. and Listeria monocytogenes in Liquid Egg Yolk and Egg White

Decimal reduction times (D values) were determined for Salmonella spp. and Listeria monocytogenes (five pooled strains per pathogen) in raw liquid egg yolk (pH 6.3) and liquid egg white (pH 8.2 versus 9.1) by using a low-volume (0.05 ml per sample) immersed sealed-glass capillary-tube procedure. For Salmonella , D values ranged from 0.087 min (at 62.2°C) to 0.28 min (at 60°C) in yolk. and from 1.00 min (at 58.3°C) to 7.99 min (at 55.1° C) in egg white (pH 8.2). For Listeria , D values ranged from 0.58 min (at 62.2°C) to 1.34 min (at 60°C) in yolk, and from 2.41 min (at 58.3°C) to 7.59 min (at 55. 1°C) in egg white (pH 9.1). Mean ZD values for Salmonella ranged from 3.54 to 4.33°C; for Listeria , ZD values ranged from 6.06 to 9.43°C. In egg white, the heat sensitivity of both pathogens was enhanced at pH 9.1, although this trend was more evident for Salmonella spp. than for L. monocytogenes over the temperature range tested. The results indicate that USDA-prescribed minimal pasteurization requirements for liquid egg yolk (equivalent to 3.9- to 22.1-D processes, on the basis of the present study) would be far more lethal to the Salmonella and L. monocytogenes strains tested than would the corresponding thermal processes for liquid egg white (equivalent to 0.7- to 2.2-D processes).

Effects of a Direct-Fed Microbial (Primalac) on Turkey Poult Performance and Susceptibility to Oral Salmonella Challenge

A study was conducted to determine 1) the effect of a dietary direct-fed microbial (DFM) on turkey poult performance, 2) the effect of a DFM on a Salmonella challenge, and 3) the effect of feed processing on the efficacy of the dietary DFM. Day-of-hatch Large White female poults were placed in 2 rooms in 2 Petersime batteries per room. Twelve pens of 7 birds each were used in each battery (24 pens per room, 336 birds total). One of 4 dietary feed treatments was assigned to each pen (6 pens per room for each diet). One room housed non-Salmonella-challenged poults, and the other room housed poults challenged with a 1-mL oral gavage of Salmonella (10(10) cfu/mL). A single batch of starter ration was split into 4 parts and used to provide 4 dietary treatments: 1) mash feed with no DFM (M), 2) mash feed with DFM (Primalac; 0.9 kg/tonne of feed, MD), 3) pelleted (20-s steam conditioning at 80 degrees C) and crumbled feed with no DFM (C), and 4) pelleted and crumbled feed with DFM (CD). Feed and deionized, distilled water were provided ad libitum. Data were collected and analyzed separately for each room. Mortality was recorded for each pen on a daily basis and totaled by week and for the 3-wk period. Individual BW and feed consumption, by pen, were measured weekly. Weekly and cumulative BW gains and feed to gain ratios (F:G) were calculated. Liver, spleen, total and lower intestinal tract weights, intestinal length, and most-probable-number Salmonella populations were determined for one randomly selected bird per pen. Feeding processed feed resulted in improved BW and F:G. Feeding the DFM improved 3-wk cumulative F:G in birds not gavaged and reduced relative intestinal weight in birds gavaged. Salmonella populations were reduced 1 log by feeding DFM. Dietary DFM improved bird performance, reduced Salmonella populations, and was not affected by feed processing.