Influence of commercial laying hen housing systems on the incidence and identification of Salmonella and Campylobacter.

The housing of laying hens is important for social, industrial, and regulatory aspects. Many studies have compared hen housing systems on the research farm, but few have fully examined commercial housing systems and management strategies. The current study compared hens housed in commercial cage-free aviary, conventional cage, and enriched colony cage systems. Environmental and eggshell pool samples were collected from selected cages/segments of the housing systems throughout the production cycle and monitored for Salmonella and Campylobacter prevalence. At 77 wk of age, 120 hens per housing system were examined for Salmonella and Campylobacter colonization in the: adrenal glands, spleen, ceca, follicles, and upper reproductive tract. All isolates detected from environmental swabs, eggshell pools, and tissues were identified for serotype. Two predominant Salmonella were detected in all samples:S.Braenderup andS.Kentucky.Campylobacter coli and C. jejuni were the only Campylobacter detected in the flocks. Across all housing systems, approximately 7% of hens were colonized with Salmonella, whereas >90% were colonized with Campylobacter Salmonella Braenderup was the isolate most frequently detected in environmental swabs (P<0.0001) and housing system impacted Salmonella spp. shedding (P<0.0001).Campylobacter jejuni was the isolate most frequently found in environmental swabs (P<0.01), while housing system impacted the prevalence of C. coli and jejuniin ceca (P<0.0001). The results of this study provide a greater understanding of the impact of hen housing systems on hen health and product safety. Additionally, producers and academia can utilize the findings to make informed decisions on hen housing and management strategies to enhance hen health and food safety.

Prevalence and Serogroup Diversity of Salmonella for Broiler Neck Skin, Whole Carcass Rinse, and Whole Carcass Enrichment Sampling Methodologies following Air or Immersion Chilling.

The purpose of this study was to evaluate neck skin (NS), whole carcass rinse (WCR), and whole carcass enrichment (WCE) sampling procedures for Salmonella isolation and serogroup identification from the same broiler chicken carcass treated with air or immersion chilling. Commercially processed and eviscerated broiler carcasses were collected from a commercial processing plant, individually bagged, and transported to the pilot processing plant. In experiment 1, carcasses were air chilled to 4°C. In experiment 2, carcasses were immersion chilled with or without chlorine. After air chilling, Salmonella was detected on 78% of NS and 89% of WCE samples. Only one Salmonella serogroup was detected from each of 13 Salmonella-positive NS samples, and two serogroups were detected on 1 Salmonella-positive NS sample. Only one Salmonella serogroup was detected from each of 13 Salmonella-positive WCE samples, and two serogroups were detected from 3 Salmonella-positive WCE samples. After immersion chilling without chlorine, Salmonella was detected on 38% of NS, 45% of WCR, and 100% of WCE samples. Without chlorine, the 15 Salmonella-positive NS samples included 14 samples with one serogroup and 1 sample with two serogroups. Only one Salmonella serogroup was detected from WCR samples after immersion chilling. Of 40 Salmonella-positive WCE samples, 23 had a one, 14 had two, and 3 had three Salmonella serogroups. After immersion chilling with chlorine, Salmonella was detected on 35% of NS, 0% of WCR, and 90% of WCE samples. With chlorine, the 14 Salmonella-positive NS samples included 11 samples with one serogroup and 3 samples with two serogroups. No Salmonella serogroups were detected from WCR samples after immersion chilling with 20 mg/liter free chlorine. The 36 Salmonella-positive WCE samples included 21 samples with one serogroup and 15 samples with two serogroups. NS and WCE sampling methodologies yielded similar prevalence and serogroup diversity after air chilling. However, after immersion chilling with or without chlorine, WCE sampling yielded significantly higher (α ≤ 0.05) prevalence and serogroup diversity than either NS or WCR sampling methodologies.

Microbiological impact of three commercial laying hen housing systems.

Hen housing for commercial egg production continues to be a societal and regulatory concern. Controlled studies have examined various aspects of egg safety, but a comprehensive assessment of commercial hen housing systems in the US has not been conducted. The current study is part of a holistic, multidisciplinary comparison of the diverse aspects of commercial conventional cage, enriched colony cage, and cage-free aviary housing systems and focuses on environmental and egg microbiology. Environmental swabs and eggshell pools were collected from all housing systems during 4 production periods. Total aerobes and coliforms were enumerated, and the prevalence of Salmonella and Campylobacter spp. was determined. Environmental aerobic and coliform counts were highest for aviary drag swabs (7.5 and 4.0 log cfu/mL, respectively) and enriched colony cage scratch pad swabs (6.8 and 3.8 log cfu/mL, respectively). Aviary floor and system wire shell pools had the greatest levels of aerobic contamination for all eggshell pools (4.9 and 4.1 log cfu/mL, respectively). Hens from all housing systems were shedding Salmonella spp. (89-100% of manure belt scraper blade swabs). The dry belt litter removal processes for all housing systems appear to affect Campylobacter spp. detection (0-41% of manure belt scraper blade swabs) considering detection of Campylobacter spp. was much higher for other environmental samples. Aviary forage area drag swabs were 100% contaminated with Campylobacter spp., whereas enriched colony cage scratch pads had a 93% positive rate. There were no differences in pathogen detection in the shell pools from the 3 housing systems. Results indicate egg safety is enhanced when hens in alternative housing systems use nest boxes. Additionally, current outcomes indicate the use of scratch pads in hen housing systems needs to be more thoroughly investigated for effects on hen health and egg safety.

Sampling naturally contaminated broiler carcasses for Salmonella by three different methods.

Postchill neck skin maceration (NSM) and whole-carcass rinsing (WCR) are frequently used methods to detect salmonellae from processed broilers. These are practical, nondestructive methods, but they are insensitive and may result in false negatives (20 to 40%). Neck skin samples comprise only 4% of the skin from the broiler carcass by weight, while WCR will not detect firmly attached Salmonella organisms and only 7.5% of the rinsate is utilized. Whole-carcass enrichment (WCE) involves incubation of the whole carcass overnight in a preenrichment broth and can recover as few as 8 inoculated Salmonella cells per carcass. The objective of this study was to use NSM, WCR, and WCE sampling to detect naturally occurring Salmonella from the same commercially processed broiler either prechill or postchill. Ten carcasses were obtained prechill and another 10 postchill on each of two replicate days from each of two commercial processing plants. From each carcass, 8.3 g of neck skin was sampled, and then the carcass was rinsed with 400 ml of 1% buffered peptone water. Thirty milliliters was removed and incubated (WCR), and the remaining 370 ml of broth and the carcass were incubated at 37°C for 24 h (WCE). Overall, Salmonella organisms were detected on 21, 24, and 32 of 40 prechill carcasses by NSM, WCR, and WCE, respectively, while 2, 2, and 19 of 40 postchill carcasses were positive by the respective methods. Prechill carcasses were 64% (77 of 120) positive for Salmonella, while postchill carcasses were 19% (23 of 120) positive. Commercial processing reduced the positive-sample prevalence by 45%. Salmonella organisms were detected on 20% (24 of 120) of the samples from plant 1 and 63% (76 of 120) of the carcasses from plant 2. This study demonstrates significant differences in the results for Salmonella prevalence among sampling methods both before and after immersion chilling, as well as between processing plants on days that samples were taken.

Comparison of shell bacteria from unwashed and washed table eggs harvested from caged laying hens and cage-free floor-housed laying hens.

These studies evaluated the bacterial level of unwashed and washed shell eggs from caged and cage-free laying hens. Hy-Line W-36 White and Hy-Line Brown laying hens were housed on all wire slats or all shavings floor systems. On the sampling days for experiments 1, 2, and 3, 20 eggs were collected from each pen for bacterial analyses. Ten of the eggs collected from each pen were washed for 1 min with a commercial egg-washing solution, whereas the remaining 10 eggs were unwashed before sampling the eggshell and shell membranes for aerobic bacteria and coliforms (experiment 1 only). In experiment 1, the aerobic plate counts (APC) of unwashed eggs produced in the shavings, slats, and caged-housing systems were 4.0, 3.6, and 3.1 log(10) cfu/mL of rinsate, respectively. Washing eggs significantly (P < 0.05) reduced APC by 1.6 log(10) cfu/mL and reduced the prevalence of coliforms by 12%. In experiment 2, unwashed eggs produced by hens in triple-deck cages from 57 to 62 wk (previously housed on shavings, slats, and cages) did not differ, with APC ranging from 0.6 to 0.8 log(10) cfu/mL. Washing eggs continued to significantly reduce APC to below 0.2 log(10) cfu/mL. In experiment 3, the APC for unwashed eggs were within 0.4 log below the APC attained for unwashed eggs in experiment 1, although hen density was 28% of that used in experiment 1. Washing eggs further lowered the APC to 0.4 to 0.7 log(10) cfu/mL, a 2.7-log reduction. These results indicate that shell bacterial levels are similar after washing for eggs from hens housed in these caged and cage-free environments. However, housing hens in cages with manure removal belts resulted in lower APC for both unwashed and washed eggs (compared with eggs from hens housed in a room with shavings, slats, and cages).

Comparison between rinse and crush-and-rub sampling for aerobic bacteria recovery from broiler hatching eggs after sanitization.

This study compared surface and deep eggshell aerobic bacteria recovered by the rinse and crush-and-rub sampling methods for commercial hatching eggs after treatment with sanitizers. Eggs were arranged into 5 treatments consisting of no treatment, water, and 3 sanitizers. The sanitizers were H(2)O(2), phenol, and Q(4)B (a compound chemical containing 4 quaternary ammoniums and 1 biguanide moiety). Eggs were sprayed according to treatment and allowed to dry for 1 h before sampling. To collect samples for the eggshell rinse, each egg was massaged in a plastic bag with 20 mL of saline. Eggshells were then aseptically opened and their contents were discarded before being individually crushed into 50-mL centrifuge tubes containing 20 mL of saline. Aerobic bacteria were enumerated on Petrifilm after 48 h of incubation at 37°C. Aerobic bacteria recovered (log(10) cfu/mL) from the eggshell rinse were highest and similar for the no-treatment (4.0) and water (3.7) groups, lower for the phenol (3.2) and H(2)O(2) (3.1) groups, and lowest for the Q(4)B (2.4) group. Aerobic bacteria levels with the crush-and-rub method were similar for the no-treatment (2.5) and water (2.3) groups, lower for the phenol (1.6) group, intermediate for the H(2)O(2) (1.2) group, and lowest for the Q(4)B (0.9) group. The overall correlation between the rinse and crush-and-rub sampling methods for individual egg aerobic bacteria counts was r = 0.71. The correlation within each treatment revealed the following r values: no treatment, 0.55; water, 0.72; H(2)O(2), 0.67; phenol, 0.73; and Q(4)B, 0.38. A second experiment was designed to further examine the lower aerobic bacterial levels recovered by the crush-and-rub method (for previously rinsed eggs) than the levels recovered in the initial eggshell rinse sample. Eggs were either rinsed and then crushed and rubbed, or they were only crushed and rubbed without a prior rinse. Results confirmed a significant decrease (1.5 log(10) cfu/mL) in bacteria levels between the initial rinse (4.4) and the subsequent crush and rub (2.9) for the same eggshell. For the crush-and-rub eggs with no previous rinsing, the bacteria recovery level (3.9) was not significantly different from levels for the rinse method. Therefore, either the rinse or crush-and-rub sampling methods can be used to recover similar levels of eggshell aerobic bacteria.

Plant factors influencing enzyme retting of fiber and seed flax.

Retting, which is the microbial activity through which bast fibers are released from nonfiber tissues, is the limiting factor in flax processing. The objective of this work is to identify chemical and structural characteristics in a variety of fiber and seed flax types that influence enzyme retting in a recently developed method. Analyses of flax retted in a series of tests, including two enzyme rettings in some cases, indicated that lignin did not limit the separation of fibers from shive and showed that pectinases in enzyme-retting mixtures could ret fiber and seed flax. However, mature stems, such as that in flax produced for seed, had greater amounts of cutin and wax in the cleaned fiber product, suggesting that the cuticle could be a greater antiquality factor in seed versus fiber flax. With seed flax, the fraction of finer fibers produced during retting was significantly lower than with fiber flax. Results indicated that enzyme retting could be used to obtain flax fibers from seed flax stem residues and add value to this agricultural material.

Identification and retting efficiencies of fungi isolated from dew-retted flax in the United States and europe.

Seven strains of filamentous fungi and one yeast were isolated from flax that was dew retted in the United States. These filamentous fungi were subcultured to purity and identified, and six appear not to have been reported earlier as isolates from dew-retted flax. Five of the purified U.S. strains, two fungi isolated from flax that was dew retted in Europe, and a laboratory culture of Aspergillus sojae were tested for their ability to ret flax stems. The monocultures were evaluated for the degree of retting, fiber strength, dry weight loss, and tactile response (i.e., feel of softness) as reflected in the retted fiber. Structural modifications of representative samples of the retted flax were assessed by scanning electron microscopy. All of the filamentous fungi were able to carry out some retting, whereas the isolated yeast could not. All organisms produced pectinases when they were cultivated in shake flasks on ball-milled flax as the sole carbon source. Some fungi also produced cellulases, mannanases, and xylanases. Rhizomucor pusillus and Fusarium lateritium were noteworthy as retting organisms by their high level of pectinase activity, ability to attack noncellulosic cell types without attacking cellulose, capacity to penetrate the cuticular surface of the stem, and efficient fiber release from the core. The results indicated that these organisms deserve further study as potential organisms for retting of bast fibers in industrial applications.

Alterations in structure, chemistry, and biodegradability of grass lignocellulose treated with the white rot fungi Ceriporiopsis subvermispora and Cyathus stercoreus.

The white rot fungi Ceriporiopsis subvermispora FP-90031-sp and Cyathus stercoreus ATCC 36910 were evaluated for their ability to delignify Bermuda grass (Cynodon dactylon) stems and improve biodegradability. Compositional and structural alterations in plant cell walls effected by the fungi were determined by nuclear magnetic resonance spectroscopy, gas chromatography of alkali-treated residues, microspectrophotometry, and electron microscopy. Contaminating bacteria and fungi, which grew from unsterilized Bermuda grass stems, did not alter the improvement in grass biodegradability by either of the fungi from that of gas-sterilized stems. The biodegradation of stems by ruminal microorganisms, after treatment for 6 weeks with C. subvermispora or C. stercoreus, was improved by 29 to 32% and by 63 to 77%, respectively; dry weight losses caused by pretreatment with the fungi were about 20% over that in untreated, control stems. Both fungi preferentially removed aromatics to carbohydrates, and C. subvermispora removed proportionately more guaiacyl units than did C. stercoreus. Substantial amounts of ester-linked p-coumaric and ferulic acids were removed by both fungi, and about 23 and 41% of total aromatics (determined after 4 M NaOH direct treatment) were removed from the plant biomass after incubation with C. subvermispora and C. stercoreus, respectively. UV absorption microspectrophotometry indicated that ester-linked phenolic acids were totally removed from the parenchyma cell walls, and these cells were readily and completely degraded by both fungi. However, aromatic constituents were only partially removed from the more recalcitrant sclerenchyma cell walls, resulting in variation in electron density and random digestion pits after incubation with fiber-degrading bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)

p-Coumaroyl and feruloyl arabinoxylans from plant cell walls as substrates for ruminal bacteria.

Growth of the ruminal bacteria Ruminococcus flavefaciens FD1, Selenomonas ruminantium HD4, and Butyrivibrio fibrisolvens 49 was limited by ester-linked feruloyl and p-coumaroyl groups. The limitation of growth on phenolic acid-carbohydrate complexes varied with individual bacteria and appeared to be influenced by ability to hydrolyze carbohydrate linkages.

Preservation of ruminal bacterium capsules by using lysine in the electron microscopy fixative.

Ruminal bacteria from axenic cultures of Ruminococcus flavefaciens FD1, Butyrivibrio fibrisolvens 49, and bacterial types from the ruminal ecosystem that were fixed with 50 mM lysine (l-lysine hydrochloride) added to glutaraldehyde had better-preserved capsules and extracellular material than bacteria fixed without lysine.

Physical degradation of lignified stem tissues by ruminal fungi.

Ruminal bacteria or fungi were selected by the addition of cycloheximide or streptomycin and penicillin, respectively, to ruminal fluid, and the weakening and degradation of lignified tissues in alfalfa and Bermuda grass stems by these treatments and whole ruminal fluid were evaluated in vitro. Dry weight loss in alfalfa was similar for whole ruminal fluid and streptomycin-penicillin treatment, whereas that with streptomycin-penicillin treatment was significantly higher (P

Mixed fungal populations and lignocellulosic tissue degradation in the bovine rumen.

Anaerobic fungi in ruminal fluid from cows eating Bermuda grass hay plus a grain and minerals supplement were evaluated for diversity in sporangial morphotypes and colony growth patterns and for the degradation of various lignocelluloses. In selective cultures containing streptomycin and penicillin, an active population of ruminal fungi colonized leaf blades and degraded fiber at rates and extents almost equal to that of the total ruminal population. Three major sporangial morphotypes were consistently observed on leaf blades: oval, globose, and fusiform. Fungal colonies representing three distinct growth types consistently developed in anaerobic roll tubes inoculated with strained ruminal fluid. Sporangial morphotypes could not be matched to colony types due to multiple sporangial forms within a colony. Under identical growth conditions, one type exhibited a monocentric growth pattern, while two types exhibited polycentric growth patterns previously unreported in ruminal fungi. Mixed ruminal fungi in selective cultures or in digesta taken directly from the rumen produced a massive clearing of the sclerenchyma. Quantitation of tissue areas in cross sections by light microscopic techniques showed that fungal incubations resulted in significant (P = 0.05) increases in sclerenchyma degradation compared to whole ruminal fluid incubations. The mestome cell wall was at times penetrated and partially degraded by fungi; the colonization was less frequent and to a lesser degree than with the sclerenchyma. Conversely, ruminal bacteria were not observed to degrade the mestome sheath. Phenolic monomers at 1 mM concentrations did not stimulate to a significant (P = 0.05) extent the dry weight loss or fungal colonization of leaf blades; at 10 mM concentrations cinnamic and benzoic acids were toxic to ruminal fungi.

Degradation of bermuda and orchard grass by species of ruminal bacteria.

Fiber degradation in Bermuda grass and orchard grass was evaluated gravimetrically and by scanning and transmission electron microscopy after incubation with pure cultures of rumen bacteria. Lachnospira multiparus D-32 was unable to degrade plant cell wall components. Butyrivibrio fibrisolvens 49 degraded 6 and 14.9% of the fiber components in Bermuda grass and orchard grass, respectively, and Ruminococcus albus 7 degraded 11.4% orchard grass fiber but none in Bermuda grass. Both B. fibrisolvens and R. albus lacked capsules, did not adhere to fiber, and degraded only portions of the more easily available plant cell walls. R. flavefaciens FD-1 was the most active fiber digester, degrading 8.2 and 55.3% of Bermuda and orchard grass fiber, respectively. The microbe had a distinct capsule and adhered to fiber, especially that which is slowly degraded, but was able to cause erosion and disorganization of the more easily digested cell walls, apparently by extracellular enzymes. Results indicated that more digestible cell walls could be partially degraded by enzymes disassociated from cellulolytic and noncellulolytic bacteria, and data were consistent with the hypothesis that the more slowly degraded plant walls required attachment. Microbial species as well as the cell wall architecture influenced the physical association with and digestion of plant fiber.

Photosynthesis, Morphology, Leaf Anatomy, and Cytogenetics of Hybrids between C(3) and C(3)/C(4)Panicum Species.

The Laxa group of the Panicum genus contains species which have CO(2) exchange and anatomical characteristics intermediate to C(3) and C(4) photosynthetic types (C(3)/C(4)), and also species characterized as C(3). Hybrids were made between two of the C(3)/C(4) species and two C(3) species. Carbon dioxide exchange and morphological, leaf anatomical, and cytogenetic characteristics of F(1) hybrids between Panicum milioides Nees. ex Trin (C(3)/C(4)) and P. laxum Mez. (C(3)), P. spathellosum Doell (C(3)/C(4)) and P. boliviense Hack. (C(3)), and P. spathellosum and P. laxum were studied. There were no consistent differences in apparent photosynthesis, although two of the three hybrids had higher net CO(2) uptake than the C(3) parent. Values of inhibition of apparent photosynthesis by 21% O(2), CO(2) loss in the light, and CO(2) compensation concentration for the hybrids were between those of the parents. All three hybrids showed leaf anatomical traits, especially organelle quantities in the bundle sheath cells, between those of their respective parents. Linear regression of CO(2) compensation concentration on the percentage of mitochondria and chloroplasts in vascular bundle sheaths of the parents and hybrids gave correlation coefficients of -0.94. This suggests that the reduction in CO(2) loss in the C(3)/C(4) species, and to a lesser degree in the F(1) hybrids, was due to development of organelles and perhaps a higher proportion of leaf photorespiration in bundle sheaths. The overall morphology of the hybrids was so different from the parents that they could be described as new taxonomic forms. The chromosomes in the hybrids were mainly unpaired or paired as bivalents indicating possible homology between some parental genomes.

Enclosure of mitochondria by chloroplasts.

In Panicum species of the Laxa group, some of which have characteristics intermediate to C(3) and C(4) photosynthesis species, some mitochondria in leaf bundle sheath cells are surrounded by chloroplasts when viewed in profile. Serial sectioning of leaves of one Laxa species, Panicum schenckii Hack, shows that these mitochondria are enclosed by chloroplasts. Complete enclosure rather than invagination also is indicated by absence of two concentric chloroplast membranes surrounding the mitochondrial profiles.