Search PubMed⌕ Search

Biomedical subjects

N A Cox

Publications and source records attributed to N A Cox.

At least 37 records · Page 2Linked to original sources

Reducing airborne pathogens, dust and Salmonella transmission in experimental hatching cabinets using an electrostatic space charge system.

Electrostatic charging of particles in enclosed spaces has been shown to be an effective means of reducing airborne dust. Dust generated during the hatching process has been strongly implicated in Salmonella transmission, which complicates the cleaning and disinfecting processes for hatchers. Following two preliminary trials in which dust reduction was measured, four trials were conducted to evaluate the effectiveness of an electrostatic space charge system (ESCS) on the levels of total aerobic bacteria (TPC), enterobacteriaceae (ENT), and Salmonella within an experimental hatching cabinet. The ESCS was placed in a hatching cabinet that was approximately 50% full of 18-d-old broiler hatching eggs. The ESCS operated continuously to generate a strong negative electrostatic charge throughout the cabinet through hatching, and dust was collected in grounded trays containing water and a degreaser. An adjacent hatching cabinet served as an untreated control. Air samples from hatchers were collected daily, and sample chicks from each hatcher were grown out to 7 d of age for cecal analysis in three of the trials. The ESCS significantly (P < 0.05) reduced TPC and ENT by 85 to 93%. Dust concentration was significantly reduced (P < 0.0001) during the preliminary trials with an average reduction of 93.6%. The number of Salmonella per gram of cecal contents in birds grown to 7 d of age was significantly (P < 0.001) reduced by an average log10 3.4 cfu/g. This ionization technology is relatively inexpensive and could be used to reduce airborne bacteria and dust within the hatching cabinet.

Air Microbiology↗

Molecular subtype analyses of Campylobacter spp. from Arkansas and California poultry operations.

Campylobacter isolates from diverse samples within broiler production and processing environments were typed by using flaA short variable region DNA sequence analysis. Sixteen flocks from four different farms representing two broiler producers in Arkansas and California were analyzed. Fourteen of the flocks (87.5%) were Campylobacter-positive; two remained negative throughout the 6-week rearing period. In general, multiple clones were present within a flock. Additionally, clones found within a flock were also present on the final product, although the diversity of Campylobacter spp. on the final product appeared to be reduced relative to that observed within the flock. Comparison of clones between flocks on the same farm revealed that some clones of Campylobacter persisted in multiple flocks. Furthermore, some clones were identified across the two farms that were under the same management. In two sampling periods, environmental isolates were positive for Campylobacter prior to flock shedding. Environmental samples associated with five additional flocks were positive for Campylobacter concomitantly with recovery of Campylobacter from the birds. Analysis of the environmental isolates that were positive prior to flock shedding demonstrated that in some instances the environmental isolates possessed genotypes identical to those of isolates originating from the flock, while in other cases the environmental isolates possessed genotypes that were distantly related to isolates obtained from the flock. Analyses of environmental isolates that tested positive concurrently with the positive isolates from the flocks demonstrated varied results; in some instances the environmental isolates possessed genotypes identical to those of isolates originating from the flock, while in other cases the environmental isolates possessed genotypes that were distantly related to isolates obtained from the flock. These data suggest that the external environment may contribute to Campylobacter contamination during poultry production and processing. However, environmental contamination with Campylobacter does not appear to be the sole contributing factor.

Animals↗

Serotype tracking of Salmonella through integrated broiler chicken operations.

The widespread presence of Salmonella in all phases of broiler chicken production and processing is well documented. However, little information is available to indicate the identity and movement of specific serotypes of Salmonella through the different phases of an integrated operation. In this study, samples were collected from the breeder farm, from the hatchery, from the previous grow-out flock, from the flock during grow-out, and from carcasses after processing. Salmonella were recovered from 6, 98, 24, 60, and 7% of the samples, respectively, in the first trial and from 7, 98, 26, 22, and 36% of the samples, respectively, in the second trial. Seven different serotypes were identified in the first trial, and 12 different serotypes were identified in the second trial. For both trials there was poor correlation between the serotypes found in the breeder farms and those found in the hatchery. This finding and the fact that similar serotypes were found in the hatchery in both trials suggests that there was an endemic population of Salmonella in the hatchery. An association between the serotypes found in the hatchery and those found on the final processed carcasses was observed in both trials. This study confirms that a successful intervention program for broiler production operations must be multifaceted, with one component being disinfection in the hatchery.

Animals↗

Serotyping and ribotyping of Salmonella using restriction enzyme PvuII.

The subtyping and identification of bacterial pathogens throughout food processing and production chains is useful to the new hazard analysis critical control point-based food safety plans. Traditional manual serotyping remains the primary means of subtyping Salmonella isolates. Molecular biology techniques, however, offer the promise of more rapid and sensitive subtyping of Salmonella. This study evaluates the potential of restriction enzyme PvuII, followed by probing with the rRNA operon from Escherichia coli, to generate serotype-specific DNA fingerprints. A total of 32 identified serotypes were found with an overall agreement in 208 of the 259 (80%) isolates tested between U.S. Department of Agriculture serotype identification and riboprint serotype identification. Many of the isolates that did not correlate were serotype identified as Salmonella Montevideo, which indicates that for this serotype, there are multiple ribotypes. When Salmonella Montevideo isolates were not included, the ribotype identification agreed with serotyping in 207 of the 231 (90%) isolates. The primary outcome of any ribotyping procedure is to give distinct ribotype patterns. This extensive poultry epidemiological study demonstrates that, in addition to ribotype patterns, the identification of isolates to known serotypes provides the investigator with additional information that can be more useful than traditional epidemiology and isolate identification studies.

Animals↗

Comparison of mucosal competitive exclusion and competitive exclusion treatment to reduce Salmonella and Campylobacter spp. colonization in broiler chickens.

Control of Salmonella spp. during the earliest phases of broiler production may provide the best opportunity to reduce human pathogens on processed broiler carcasses. Application of the "Nurmi concept" has been demonstrated to be an effective means in reducing Salmonella colonization among broiler chicks. In 1989, Aho et al. developed a competitive exclusion (CE) culture for control of Salmonella spp., whereas a mucosal competitive exclusion culture (MCE) developed in the United States was originally created to control Campylobacter colonization (Stern et al., 1995). The major differences in the two patents were the higher level of anaerobic culture required, the degree of epithelial scraping and washing of the ceca, media used for subculturing, and the culture incubation temperatures (35 C vs. 42 C). The CE and MCE were compared for efficacy in reducing Salmonella and Campylobacter colonization in broiler chicks. Nine adult birds (three for each of three replicates) were slaughtered, and each of a bird's paired ceca were used to produce corresponding antagonistic microflora, which were administered to day-of-hatch chicks. The chicks (a total of 210) were challenged 24 h later with Salmonella and Campylobacter and were killed 1 wk later, and levels of the pathogens were determined. Ninety CE-treated birds were significantly more colonized by Salmonella typhimurium than those 90 chicks treated with the MCE microflora (3.97 log 10 cfu/g cecal contents vs. 1.25 log 10 cfu/g cecal contents). Also, Campylobacter spp. colonization of these birds was significantly higher for CE-treated birds when compared with MCE-treated birds (6.96 log 10 cfu/g cecal contents vs. 5.03 log 10 cfu/g cecal contents). These results can be useful in developing intervention strategies to reduce chicken colonization by Salmonella and Campylobacter.

Animals↗

Detection of Campylobacter spp. in ceca and crops with and without enrichment.

The purpose of this experiment was to determine how sampling method (direct plating or enrichment) affected the rate of Campylobacter spp. isolation from crop and cecal samples. In four separate trials, 32 New York-dressed broiler carcasses were obtained from commercial plants (n = 128). Crops and ceca were removed aseptically, direct plated, and enriched. Samples were direct-plated on Campy-Cefex plates that were incubated at 42 C for 36 to 48 h under a microaerobic atmosphere (5% O2, 10% CO2, 85% N2). After direct plating, samples were enriched in Bolton broth at 37 C for 4 h and 42 C for 20 h under a microaerobic atmosphere before plating onto Campy-Cefex plates. Campylobacter spp. was detected in 95.3% of direct-plated crop samples and 99.2% of enriched crop samples. Campylobacter spp. was detected in 100% of direct-plated cecal samples and 63.3% of enriched cecal samples. All 128 crop and cecal samples were positive for the organism by one or both methods. Mean counts of Campylobacter spp. were 3.6 log10 cfu/g of crop sample and 6.8 log10 cfu/g of cecal sample. For these two sample types, both of which tend to be contaminated with many viable cells, direct plating is sufficient for isolation of Campylobacter. Direct plating also provides an estimate of contamination level. Enrichment of cecal samples resulted in a decreased rate of detection and did not allow estimation of numbers of Campylobacter. The large numbers of non-Campylobacter species that inhabit the intestinal tract may out-compete Campylobacter during enrichment, confounding detection.

Animals↗

Sources and movement of Salmonella through integrated poultry operations: a multistate epidemiological investigation.

The prevalence of Salmonella from numerous sources in 32 integrated broiler operations of high- and low-performing broiler houses was characterized from four states across four seasons. Previous studies of Salmonella in broilers have been limited in scope, offering only a snapshot of pathogen prevalence as seen on a small number of individual farms. Twenty-six different sample types were collected from the hatchery to the end of processing, and Salmonella was found in all sample types. A total of 10,740 samples were analyzed for Salmonella, and 973 (9.1%) of these samples, including 49 of 798 (6.1%) carcass rinse samples, were Salmonella positive. Hatchery transport pads (389 of 765, 50.8%), flies (28 of 150, 18.7%), drag swabs (57 of 402, 14.2%), and boot swabs (20 of 167, 12%) were samples from which Salmonella was most frequently isolated. Thirty-six different serotypes were identified, and the most frequently encountered serotypes were Salmonella Senftenberg, Salmonella Thompson, and Salmonella Montevideo. Determining critical contaminating sources and following the movement of Salmonella through integrated poultry operations will help researchers and the industry develop practical intervention strategies.

Animals↗

Distribution of Campylobacter spp. in selected U.S. poultry production and processing operations.

A study was conducted of 32 broiler flocks on eight different farms, belonging to four major U.S. producers. The farms were studied over I complete calendar year. Overall, 28 (87.5%) of the flocks became Campylobacter positive, and only four (12.5%) remained negative throughout the 6- to 8-week rearing period. In the majority of flocks, sampled every 2 weeks throughout production, Campylobacter-positive fecal and cecal samples were not detected until 4 to 8 weeks of age. In only six of the flocks were environmental samples found to be positive before shedding of Campylobacter was detected in the birds. Even in some of the Campylobacter-negative flocks, contamination of the rearing environment was positive for Campylobacter but did not result in the birds subsequently excreting the organism. These findings are discussed in relation to U.S. husbandry practices and present uncertainty about sources of Campylobacter infection for poultry flocks. Birds were often transported to the processing plant in coops that were already contaminated with Campylobacter, and the organisms were sometimes found in samples of scald water and chill water. After chilling, the proportions of Campylobacter-positive carcasses from different producers ranged from 21.0 to 40.9%, which is lower than in other studies, and possible reasons are considered.

Age Factors↗

Difficulty in recovering inoculated Campylobacter jejuni from dry poultry-associated samples.

We inoculated 5 cm2 of clean chick pads, 5 g of clean pine shavings, and fresh unsanitized broiler breeder eggshell halves with a cell suspension of Campylobacter jejuni in physiological saline. Inoculation levels were 10(2), 10(3), or 10(4) cells per sample. The samples were allowed to remain at room temperature for 15, 30, or 60 min before addition of enrichment broth. When chick pad samples were inoculated with 102 cells, by 15 min 40% of the samples had detectable levels of Campylobacter, and by 30 to 60 min Campylobacter could be detected in only 20% of the samples. With samples of pine shavings, only 25% of those inoculated with 103 cells were positive for Campylobacter after 15 min and only 5% were positive for Campylobacter after 30 min. When 104 cells were inoculated onto litter, Campylobacter was recovered from 20% of the samples at 15 min and 15% of the samples after 30 min. Eggshells were also found to be a harsh environment. When the inoculum was 102 at 15 min, 8 of 10 samples were positive for Campylobacter but at 60 min only 10% of the samples remained positive for Campylobacter. The current cultural methods may not be adequate for recovering low numbers of Campylobacter from dry samples. Campylobacter may be present but culturally undetectable in the commercial hatchery and hatchery environment.

Animals↗

Salmonella penetration of egg shells and proliferation in broiler hatching eggs--a review.

The presence of salmonellae in fertile broiler hatching eggs has been clearly identified as a critical control point in the salmonellae contamination of broiler chickens. This paper reviews the published research studies on a) the penetration and proliferation of salmonellae in hatching eggs, b) the consequences of this contamination on the contamination of the final product, and c) the egg's defenses against invading salmonellae. A better understanding of the material in this review paper will assist poultry researchers and the poultry industry in continuing to make progress in reducing and eliminating salmonellae from fertile hatching eggs, hatcheries, and breeder flocks.

Animals↗

Efficacy of an herbal extract on the microbiological quality of broiler carcasses during a simulated chill.

Protecta II, an herbal extract on an NaCl carrier, was evaluated in a 30-min, 1 C simulated chill for its effectiveness of lowering microbial counts on broiler carcasses. Eighteen broiler carcasses were obtained from a local processing plant after final wash but before chill, placed into an insulated container, and transported to the research facility for treatment. Six plant run controls (PRC) were immediately bagged on return to the pilot plant, and a whole-carcass rinse was performed. The remaining carcasses were subjected to a 30-min chill (1 C) in tap water or a 2% solution of Protecta II, (n = 6 per treatment). After treatment, carcasses were rinsed with tap water and subjected to the whole-carcass rinse procedure. All rinse diluents were microbiologically analyzed for total aerobes, coliforms, generic Escherichia coli, and Campylobacter. Six replications were analyzed on 6 different d for a total 36 carcasses per treatment and 36 PRC. The PRC carcasses had 3.7, 2.5, 2.1, and 2.0 log10 cfu/mL for total aerobes, coliforms, generic E. coli, and Campylobacter. Water treatment significantly reduced counts (2.6, 1.4, 0.7, and 0.9 log10 cfu/mL, respectively) when compared with the PRC. Protecta II treatment significantly reduced counts (P < 0.01) even further to counts of 0.06, 0.04, 0.01, and 0.00 log10 cfu/mL for total aerobes, coliforms, Campylobacter, and E. coli, respectively. Detectable levels of the monitored organisms were 1 cell/mL (log10 0) for the E. coli, coliforms, and total counts and 10 cells/mL (log10 1) for the Campylobacter. Microbial counts for carcasses treated with Protecta II would be considered too low to be detected (<1 cell/mL).

Animals↗

Commercial field trial evaluation of mucosal starter culture to reduce Salmonella incidence in processed broiler carcasses.

A series of four paired-house studies was conducted in Arkansas, Alabama, and Georgia (two farms) to determine the efficacy of Mucosal Starter Culture (MSC) in eliminating or reducing salmonellae in broiler chickens. Randomly designated chicks were treated twice with MSC. First they were sprayed with an MSC solution using a spray vaccination cabinet in the hatchery, and then they received MSC in the first drinking water at the growing house. Chicks were grown in identically constructed and equipped paired houses managed by the same grower. At the end of grow-out, broilers were tested for the presence of salmonellae on the farm and during processing. In three trials where no hatchery salmonellae were found, less salmonellae were found on MSC-treated chickens compared to untreated chickens. On the farm at the end of grow-out, salmonellae were detected in 54 of 150 untreated control chickens compared to 40 of 180 MSC-treated chickens. In the processing plant, significantly (P < or = 0.05) more salmonellae were detected on prechill untreated control carcasses (23 of 180) compared to MSC-treated carcasses (12 of 180) and on untreated postchill processed carcasses (9 of 180) compared to MSC-treated carcasses (0 of 180). In one trial where appreciable (28% of egg shell samples) salmonellae was found before treatment with the MSC, more salmonellae were found in the treated birds than in the control birds both on the farm and after processing. These data confirm that when salmonellae levels were controlled in the hatchery, a significant reduction in the salmonellae was found on processed broiler carcasses treated with MSC and that this reduction in salmonellae was carried through processing to the final processed carcass, thus potentially reducing consumer exposure to salmonellae.

Animals↗

Eggshell membrane structure and penetration by Salmonella typhimurium.

Eggshell membrane was removed from 10 broiler hatching eggs at approximately monthly intervals through the productive life of a commercial flock. A piece of membrane (2 by 2 cm) was used to cover an opening in an apparatus designed to test penetration by Salmonella Typhimurium. The membrane was placed between two chambers as the only means of liquid transfer. The chamber above the membrane was filled with a suspension of Salmonella Typhimurium cells. Sampling of the bottom chamber provided cultural evidence of the penetration by a marker strain of Salmonella Typhimurium. Samples were drawn at 15 min and at 1, 3, 5, 7, 12, 24, and 48 h. Following the cultural penetration experiment, the same pieces of membrane were removed and stained for microscopic examination. Membrane structure was examined with the use of a confocal laser scanning microscope. Each image consisted of a composite of eight 1-microm optical slices showing the fibers making up the outer surface of the membrane. These images were transferred to an image analysis software package that allowed the measurement of the interfiber area. No clear correlation could be made between the average size of all interfiber areas or the total interfiber area measured in the outer 8 microm of the outer membrane and the ability of Salmonella Typhimurium to penetrate the same piece of eggshell membrane.

Animals↗

Mucosal competitive exclusion to reduce Salmonella in swine.

A mucosal competitive exclusion culture has been shown to reduce or eliminate Salmonella spp. in poultry. Using similar techniques, a mucosal competitive exclusion culture from swine (MCES) was produced from the cecum of a 6-week-old pig. Suckling pigs were inoculated with 5 ml of MCES by oral gavage within 6 h postfarrowing (PF) and again at 24 h PE All pigs were challenged with 10(3) CFU of Salmonella Choleraesuis at 48 h PF by intranasal instillation, including pigs from two sows that had not been given MCES. Clinical signs and rectal swabs were monitored daily, and pigs were allowed to suckle throughout the experiment. All pigs underwent necropsy on day 7 PF, and presence of Salmonella was determined in both qualitative (10 tissues) and quantitative (two tissues) samples. Clinical signs were inapparent in all pigs throughout the experiment. Recovery of Salmonella from rectal swabs was variable. However, 28% of the gut tissues were positive from the MCES-treated pigs versus 79% from the control pigs. A 2- to 5-log10 reduction of Salmonella in the cecal contents or ileocolic junction was observed in the MCES-treated pigs when compared with the controls. These data indicate that use of MCES may be a useful approach for control of Salmonella.

Animals↗

Effect of Salmonella in young chicks on competitive exclusion treatment.

The potential of Salmonella contamination in hatching cabinets to 1) generate seeder chicks and 2) interfere with the efficacy of competitive exclusion (CE) treatments was investigated in six experiments. Hatchery-generated seeder: chicks were produced by hatching in the same hatcher with inoculated eggs (immersed in a 1.0 x 10(5) cfu/mL suspension of Salmonella typhimurium or inoculated with 10(5) to 10(6) cells on the air cell membrane). Salmonella spread through the hatching cabinet to chicks hatching from uninoculated eggs in adjacent trays. In two experiments, Salmonella was isolated from 31 and 100% of the chick rinses after the birds were held in groups for 3 d in isolation units. When these hatchery-generated seeder chicks were stocked in floor pens at a 1:10 ratio with uncontaminated contact chicks, the pen environment became contaminated to the extent that greater than 50% of the contact chicks became contaminated. In two experiments with only one to three inoculated eggs per 200 egg hatching cabinet, 98% of uninoculated chicks were intestinally colonized with Salmonella after the birds were held 1 wk in isolation cabinets. This hatchery-acquired Salmonella substantially reduced the effectiveness of subsequent CE treatments to prevent Salmonella colonization of the young chicks. These studies demonstrate that control of Salmonella in hatching cabinets is critically important for control of Salmonella in broilers.

Animals↗

Effect of yeast-supplemented feed on Salmonella and Campylobacter populations in broilers.

The effect of the yeast, Saccharomyces boulardii, on experimental cecal colonization of broilers with Salmonella typhimurium and Campylobacter jejuni was investigated. Duplicate pens of broiler chicks were given ad libitum access to a standard feed supplemented with no yeast (control), or 1 g (1x), or 100 g (100x) dried S. boulardii/kg feed. All chicks except negative controls were challenged on Day 4 with 3.2 x 10(8) cfu S. typhimurium and 6.5 x 10(8) cfu C. jejuni by oral gavage. After 3 wk, the broilers were euthanatized and ceca were aseptically removed and analyzed for Salmonella and Campylobacter. Frequency of Salmonella colonization was significantly (P < 0.05) reduced due to yeast treatment. Of the positive control birds, 70% were colonized with Salmonella; whereas only 20 and 5% of the 1x and 100x yeast-treated birds were colonized. Mean number of Salmonella per gram of ceca and contents were log 1.64, 0.35, and 0.15, respectively, for the control, 1x, and 100x yeast-treated birds. Campylobacter colonization was not significantly affected by yeast treatment. Similar results were obtained from a second trial conducted in larger isolation floor pens.

Animal Feed↗

Eggshell characteristics and penetration by Salmonella through the productive life of a broiler breeder flock.

Egg weight, specific gravity, conductance, and ability of Salmonella to penetrate the shell and membranes were determined for hatching eggs from a commercial broiler breeder flock. Thirty unsanitized eggs were sampled on Weeks 29, 34, 39, 42, 48, 52, and 56 of flock age for specific gravity and conductance. An additional 10 intact eggs were inoculated with Salmonella by a temperature differential immersion method for 1 min. Eggs were then emptied of contents and filled with a selective medium that allowed visualization of Salmonella growth on the inside of the shell and membrane complex. Over the 27-wk sampling period, egg weight increased from 56 to 66 g and was positively correlated with hen age (r = 0.96, P < 0.05). However, neither specific gravity (ranging from 1.077 to 1.082) nor eggshell conductance (ranging from 14.7 to 17.9 mg weight loss/d per torr) showed any clear trend throughout the life of the flock despite the increase in egg weight. Conductance values were not correlated with specific gravity. The number of eggs positive for Salmonella penetration after 24 h incubation showed a general upward trend with flock age; however, penetration frequency and hen age were not found to be significantly correlated (P > 0.05). No relationship was found between egg specific gravity, conductance, or egg weight and the likelihood of Salmonella to penetrate the eggshell. Because shell characteristics did not change over time and the penetration patterns did vary, it is likely that factors other than specific gravity and conductance were involved in the penetration of eggshells by Salmonella.

Animals↗

Effect of cloacal plugging on microbial recovery from partially processed broilers.

Experiments were performed to test the contribution of bacteria contained in the intestinal tract of broilers at the beginning of processing to counts on the exterior of modified New York-dressed carcasses. Thirty-two birds were processed for each of seven replications. Within each replication, batches of four birds were electrocuted, scalded, and picked, with batches alternating between treatment and control groups. Treated birds were cloacally plugged with rayon fiber tampons prior to electrocution to prevent escape of intestinal contents during scalding and picking. Control birds were processed in the same manner, except that cloacal plugs were inserted immediately after defeathering to reduce escape of intestinal contents during sampling. Gram-negative enteric bacteria and Campylobacter spp. were enumerated on carcasses by whole carcass rinse procedure and in cecal contents. Counts were converted to log10 and subjected to analysis of variance. Cecal levels of Gram-negative enterics were significantly higher for plugged birds, but there was not a significant difference between levels of cecal Campylobacter spp. between treatment groups. Plugging before electrocution resulted in significantly lower levels (2.5 vs 3.0 log10 cfu/mL) of Campylobacter spp. and Gram-negative enteric bacteria (3.0 vs 3.4 log10 cfu/mL) in carcass rinses of treatment birds than in those of controls. All carcasses were positive for Gram-negative enterics. Cloacal plugging resulted in significantly lower incidence of Campylobacter spp. carcass contamination as determined by chi-square. Intestinal carriage of both campylobacters and Gram-negative enteric bacteria appears to influence the microbial quality of the carcass during processing.

Analysis of Variance↗