Vertical Stratification of Ammonia in a Broiler House

The broiler industry is not immune to the problematic nature of NH₃ production in animal rearing facilities. Though the headlines of today focus on environmental impact considerations, the detrimental effects of the house air quality on farmers and birds remain considerable for industry viability. This research investigated the vertical stratification of in-house NH₃ combined with sampling position down the center of the house and with different NH₃ analysis technologies. The results indicated that reuse of litter and house ventilation correlate to trends in NH₃ concentration at particular measurement heights. When tunnel ventilation was primary, NH₃ concentrations decreased vertically with increasing distance from the litter surface. However, with lower outside temperatures, little ventilation, and a stagnant house atmosphere, no concentration gradient was evident. The work also demonstrated significant variability among professionally calibrated instruments and traditionally used pull tubes. Characterization of interior air quality of broiler houses should consider sampling height to effectively address bird exposure.     

Risk Factors Associated with Detection of Salmonella in Broiler Litter at the Time of New Flock Placement

In this study, we investigated risk factors associated with the probability to detect Salmonella in samples of litter collected within 2 h prior to new flock placement in 76 grow‐out houses on 38 conventional broiler farms located in the US states of Mississippi, Alabama and Texas. We evaluated characteristics of location and layout of the farm; area adjacent to and surrounding the house; house construction; condition and type of equipment in the house; litter management and other production, sanitation, visitation and biosecurity practices; non‐broiler animal species on the farm; and weather conditions on the 3 days leading up to flock placement. Logistic regression was used to model the relationships between probability to detect Salmonella in litter and potential risk factors. In the screening process, each risk factor was evaluated as a single fixed effects factor in a multilevel model that accounted for variability among the sampled farms and their production complexes and companies. Of almost 370 risk factors screened, 24 were associated with the probability to detect Salmonella in litter. These were characteristics of the surroundings of the house, house construction and conditions, litter management, length of downtimes between flocks in the house, biosecurity and farm location. After investigation of collinearity between these variables and building of models for important risk factor categories, the list of candidate variables for the final model was refined to eight factors. The final model demonstrated that a higher probability of detecting Salmonella in litter was strongly associated with the use of wood to construct the base of the walls or to cover the inside of the broiler house foundation, and with the use of fresh wood shavings to top‐dress or completely replace the litter between flocks.     

Impact of added sand on the recovery of Salmonella,Campylobacter, Escherichia coli,and coliforms from prechill and postchill commercial broiler carcass halves

Salmonella and Campylobacter are often associated with raw poultry products and continue to be leading causes of food-borne gastroenteritis in the United States. As a result, the presence of these organisms on broiler carcasses is monitored on a routine basis. Abrasive rinsing methods (e.g., adding glass beads) have been shown to increase the level of bacteria recovered from carcasses or carcass parts. The objective of this study was to evaluate the addition of sand to the rinse on bacterial enumeration and the prevalence of Salmonella and Campylobacter recovered from broiler carcasses. During each of 4 replications, 6 prechill and 6 postchill broiler carcasses were collected from a commercial processing plant. All carcasses were split along the dorso-ventral midline. Carcass halves were rinsed in buffered peptone water, whereas the companion half was rinsed in buffered peptone water with sterile sand added. All carcass halves were rinsed for 1 min and the rinsate was collected. Salmonella, coliforms, and Escherichia coli were enumerated and the prevalence of Salmonella and Campylobacter was determined. Salmonella and Campylobacter were isolated from 17 and 50% of the carcass halves, respectively. There was no significant (P > 0.05) difference in Salmonella or Campylobacter prevalence from carcass halves rinsed with or without sand. The addition of sand to the rinse had no effect on the number of Salmonella, coliforms, or E. coli recovered from prechill or postchill carcass halves. These results show that adding sand to the rinse liquid did not improve the recovery of bacteria present on the carcass in either moderate (2.6 log₁₀ cfu/mL rinsate) or low numbers (<3 cfu/mL of rinsate).     

Proof of concept for eradication of vancomycin resistant Enterococcus faecium from broiler farms

Background

Vancomycin resistant enterococci (VRE) in Swedish broiler production has been shown to persist at farms between batches. The aim of this study was therefore to determine the possibility to eliminate VRE by disinfection of compartments in broiler houses as a proof of concept.

Findings

VRE could not be detected in environmental samples from the disinfected test compartments in the broiler houses but was detected in environmental samples from the control compartments. The proportion of broilers colonized with VRE decreased in both the test and the control compartments.

Conclusions

The results are promising and show that the occurrence of VRE in broiler houses can be reduced by adequate cleaning and disinfection with a combination of steam and formaldehyde.     

Detection of multidrug resistant Escherichia coli in the ovaries of healthy broiler breeders with emphasis on extended-spectrum β-lactamases producers

In the last few years, antimicrobial resistant (AMR) Escherichia coli have been detected in newborn chickens suggesting their vertical transmission from breeding birds to their offspring. However, little is known about the presence of AMR E. coli in the reproductive organs of broiler breeders. The aim of this study was to investigate the presence of E. coli in the ovaries of healthy broiler breeders and to study their antimicrobial resistance. Samples from broiler breeders (n = 80) collected from 80 different broiler breeder flocks were included in this study. Antibiotic susceptibility testing was performed using disk diffusion method according to Clinical and Laboratory Standards Institute guidelines. Minimal inhibitory concentrations (MICs) of five antimicrobial agents were determined by Etest. PCR and sequencing were used to detect the bla_ESBL genes. E. coli were detected in the ovaries of thirty seven out of 80 (46.25%) sampled flocks. High levels of resistance to various first-line antimicrobial agents were recorded in E. coli isolates. This study showed that 89.18% of E. coli isolates were multidrug resistant (MDR). Furthermore, MDR extended-spectrum β-lactamases (ESBL)-producing E. coli were detected in the ovaries of four different broiler breeder flocks. Molecular characterization revealed that three isolates harboured bla_CTX-M-1 gene and one isolate expressed bla_SHV-12 gene. In addition, one bla_CTX-M-1 -producing E. coli co-harboured the bla_TEM-1 gene. These findings would contribute to a better epidemiological understanding of MDR E. coli for improve existing preventive strategies in order to reduce the dissemination of antimicrobial resistance in the broiler production system.     

On-line brush and spray washers to lower numbers of Campylobacter and Escherichia coli and presence of Salmonella on broiler carcasses during processing

It is unclear how effective different types of broiler carcass wash steps are in lowering the presence or numbers of pathogenic bacteria. We tested for individual and combined effectiveness of 5 separate on-line wash steps applied between bleed-out and chilling in a commercial broiler processing plant. Carcasses were sampled directly before and after each wash step: pre-scald brush washer, post-feather pick (New York dressed) spray washer, inside/outside spray washer, postevisceration brush washer, and final prechill spray washer. Carcasses were examined for numbers of Campylobacter and Escherichia coli and presence of Salmonella using standard cultural methods. Overall, numbers of Campylobacter were lowered from log 2.58 to 1.15 cfu/mL of carcass rinse, but no single wash step caused a significant decrease. Overall, Salmonella prevalence was decreased from 80 to 24%; however, no wash step caused a significant decrease by itself. The 5 wash steps in series lowered E. coli numbers from log 4.60 to 2.69 cfu/mL; the New York-dressed spray wash and the postevisceration brush washer each had a significant effect on E. coli. When examined separately, the benefit of broiler carcass wash steps may not be evident. However, when combined with overall processing, wash steps can be effective to lessen bacterial contamination on carcasses and be useful for pathogen control. Additional studies are necessary to maximize the effectiveness of carcass washers.     

Risk factors for Salmonella persistence after cleansing and disinfection in French broiler-chicken houses

A prospective survey was carried out in 86 broiler houses in western France to identify risk factors for Salmonella persistence in French broiler houses. The Salmonella status of the house after cleansing and disinfection, was assessed from gauze-swab samples taken from the walls, feeders, ventilation system and bedding (analysed with classical bacteriological methods). Thirty three (38%) houses had at least one contaminated sample and were classified as Salmonella-contaminated houses in the logistic regression. The absence of a terminal disinfection and a disinfection procedure performed by the farm staff rather than a contractor were positively related to the Salmonella contamination of the house after cleansing and disinfection. The risk for Salmonella persistence after decontamination was increased if rodents were observed by the farmer, if a large part of the access area to the house was accessible to trucks, and if a disease leading to a treatment occurred in the previous flock.     

Persistence of cellulitis-associated Escherichia coli DNA fingerprints in successive broiler chicken flocks

Avian cellulitis in broiler chickens is primarily caused by Escherichia coli. Previous research found that the E. coli isolates of cellulitis origin were unique to each ranch, suggesting that these E. coli were endemic within the ranch environment. To test the hypothesis that the E. coli associated with cellulitis are endemic in the litter of the broiler house, we designed a study to determine whether E. coli DNA fingerprints associated with cellulitis persist over successive flocks that are grown in the same house. In addition, we assessed the impact of different cleaning and disinfection strategies on this persistence. Two broiler houses were followed on each of five farms over 3-4 flocks. A total of 353 E. coli isolates from cellulitis lesions were analyzed in this study, and 314 of these isolates (89%) were DNA fingerprinted by PFGE. In each ranch, there were several DNA fingerprint patterns that were present over successive flocks, regardless of the cleaning and disinfection strategy utilized. Isolates persisted as long as 191 days, implying that these E. coli are capable of persisting in the broiler house environment for long periods of time. In addition, these E. coli isolates were associated with cellulitis lesions in successive flocks. Thus, the isolates of E. coli that are associated with cellulitis in broiler chickens appear to be endemic in the litter environment of the broiler house.     

A systematic review-meta-analysis of chilling interventions and a meta-regression of various processing interventions for Salmonella contamination of chicken

Context

The results of individual studies investigating the efficacy of chilling and other processing interventions on Salmonella prevalence or concentration in broiler chicken carcasses are inconsistent or contradictory.

Objective

Determine efficacy of chilling on reducing Salmonella prevalence or concentration on broiler carcasses using systematic review-meta-analysis, and explore sources of heterogeneity among studies investigating various processing interventions through meta-regression.

Data sources

A comprehensive search included electronic search in six databases, manual search of reference lists of topic-related articles, and consultation with five topic experts to assure that all relevant intervention research was identified.

Study inclusion

Primary intervention research, published in English, encompassing control, challenge, cohort, or before-and-after study designs investigating the efficacy of any chilling or other processing interventions on Salmonella prevalence or concentration in broiler chicken carcasses.

Risk of bias assessment and data extraction

Data pertaining to study methodology and reported results, chilling or other processing intervention parameters, populations sampled and outcomes measured were assessed for methodological soundness and extracted by two independent reviewers using pretested checklists.

Results

Random-effects meta-analyses of immersion chilling with chlorine (n = 9 trials), acetic acid (n = 16) and potable water (n = 13) trended towards reductions in the odds or log₁₀ CFU/ml of Salmonella. Significant heterogeneity (P-value ≤ 0.1 and I² > 25%) precluded the reporting of pooled summary effect estimates. Meta-regression of all processing interventions indicated that serotype, disinfectant type and treatment time and pH were significantly associated with studies reporting reductions in concentration while study design, population sampled, study setting, publication date, intervention and disinfectant type, and treatment pH were significantly associated with studies reporting reductions in prevalence. Methodological and reporting flaws were consistently observed in relevant intervention research as well as a lack of studies conducted under commercial conditions and using Salmonella concentration outcomes.

Conclusions

Chilling may be effective at reducing Salmonella concentration and prevalence, but significant heterogeneity precluded reporting of pooled summary effect estimates for many chilling interventions. Investigations into potential sources of heterogeneity among all processing interventions found that the use of other chemical disinfectants, such as organic acids and surfactants might result in larger reductions in Salmonella contamination than more commonly utilized oxidizing agents like chlorine.     

Antibiogram,virulotyping and genetic diversity of Escherichia coli and Salmonella serovars isolated from diarrheic calves and calf handlers

Antimicrobial resistance profile of E. coli and Salmonella serovars isolated from diarrheic calves and handlers in Egypt is unknown due to the absence of monitoring. Therefore, this study aimed to determine the virulence, genetic and antimicrobial resistance profiles of E. coli and Salmonella serovars associated with diarrhea in calves and handlers in intensive dairy farms in Egypt. A total of 36 bacterial strains (20 E. coli and 16 Salmonella) were isolated from fecal samples of 80 diarrheic Holstein dairy calves (10 E. coli and 13 Salmonella) and hand swabs of 35 handlers (10 E. coli and 3 Salmonella) in two intensive dairy farms in Sharkia Governate in Egypt. E. coli strains belonged to six different serogroups and O114:K90 was the most prevalent serogroup (30%). However, Salmonella strains were serotyped into four different serogroups and S. Kiel was the most prevalent serotype (50%). Thirteen (65%) E. coli isolates were harbouring either stx2, eaeA and/or astA virulence-associated genes. However, stn and spvC virulence genes were detected in 2 (12.5%) and 4 (25%) of Salmonella isolates, respectively. E. coli isolates showed marked resistance to ampicillin (75%), while Salmonella strains exhibited high resistance to amikacin (100%), gentamicin (93.75%) and tobramycin (87.5%). Results of the present study showed that E. coli and Salmonella serovars isolated from diarrheic calves and handlers in intensive dairy farms in Egypt exhibited resistance to multiple classes of antimicrobials, which may pose a public health hazard. Thus, the continuous monitoring of antimicrobial resistance is necessary for both humans and veterinary medicine to decrease the economic losses caused by antimicrobial-resistant strains in animals as well as the zoonotic risk.     

A Longitudinal Study of Salmonella Infection in Different Types of Turkey Flocks in Great Britain

Salmonella is, after Campylobacter, the most reported zoonotic pathogen in the EU. Poultry are a common source of infection to humans, and turkey flocks are commonly colonized with the organism. We investigated the prevalence and risk factors of Salmonella infection in 179 houses in 60 holdings representative of turkey meat and breeder production in Great Britain. From each holding, up to four houses were chosen, and two consecutive flocks per house were sampled/tested for Salmonella to investigate the persistence, elimination and introduction of Salmonella in consecutive crops. At the first sampling, the overall flock‐level Salmonella prevalence was 32.8% and 8.9% for meat and breeding flocks respectively. There was a higher prevalence of Salmonella in flocks in the rearing stage than in the fattening and breeding stages. At the first sampling, the flock‐level prevalence of Salmonella was 26.8% (95% CI: 20.7–33.7%), while the prevalence level in the subsequent flock was 20.5% (95% CI: 13.6–29.7%). No houses were positive for any of the EU‐regulated serovars. The most commonly encountered serovars were S. Kottbus and S. Kedougou. Carry‐over of infection was observed in 44.8% of the positive houses, and introduction of new infection occurred in 8.4% of houses. Data from the questionnaires and auditing of all holdings and houses were combined and used to calculate adjusted farm‐ and house‐adjusted risk factors. Significant risk factors were feed from a source other than a national compounder (OR = 2.4), feeder type other than pan feeders (OR = 2.4) and hygiene practices other than terminal cleaning and disinfection using power‐washing with sanitizer and anteroom with boot change (OR = 2.8). The study discusses the main challenges currently faced by the industry to control Salmonella in turkey production.     

Occurrence of foodborne bacteria in Alberta feedlots

The occurrence of generic Escherichia coli, E. coli O157, Salmonella, and Campylobacter in cattle manure, beef carcasses, catch basin water, and soils receiving manure application was determined in 21 Alberta feedlots. In cattle manure, generic E. coli (98%, 2069/2100) and Campylobacter (76%, 1590/2100) were frequently detected; E. coli O157 (7%, 143/2100) and Salmonella (1%, 20/2100) were less frequently detected. Samples from beef carcasses in the cooler following Hazard Analysis Critical Control Point interventions yielded only 1 isolate each of generic E. coli and Campylobacter (1/1653) and no Salmonella (0/1653). Catch basin water specimens were positive for generic E. coli in both the spring (62%, 13/21) and the fall (52%, 11/21). Other bacteria were detected only in the spring water specimens, including E. coli O157 (29%, 6/21), Salmonella (5%, 1/21), and Campylobacter (52%, 11/21). Generic E. coli was frequently isolated from soil specimens (30%, 27/88), but E. coli O157 was not found in soil samples obtained in the spring and was only occasionally detected in the fall samples (9%, 3/32). Salmonella were occasionally found in the soil specimens collected in the spring (3%, 2/56), but not in the fall season (0/32). Campylobacter jejuni was frequent in cattle manure (66%, 1070/1623), but rare in carcass and environmental samples. E. coli O157 and Salmonella were rarely detected in cattle or the environment. Generic E. coli and Salmonella were rarely detected on carcasses.     

Studies on Salmonella from Floor Litter of 60 Broiler Chicken Houses in Nova Scotia

Fifteen floor litter samples were collected from each of 60 broiler chicken houses in Nova Scotia. Salmonella was recovered from 270 litter samples (30%) in 33 of 60 (55%) houses. All isolates were serotyped. Salmonella heidelberg and S. infantis were the most common of the 13 serotypes found. Isolates of selected serotypes were phage typed. Most isolates were susceptible to all common antimicrobial agents.     

Spread of extended-spectrum beta-lactamase producing Escherichia coli isolates in Swedish broilers mediated by an incI plasmid carrying blaCTX-M-1

Background

The already high and increasing occurrence of extended-spectrum beta-lactamases (ESBL) producing Escherichia coli in European broiler populations is of concern due to the fact that third and fourth generation cephalosporins are deemed critically important in human medicine. In Sweden 34% of the broilers carry ESBL/pAmpC producing E. coli in their gut, despite the absence of a known selection pressure such as antimicrobial usages. The aim of the current study was to characterise a selection of E. coli strains carrying the bla_CTX-M-1, to determine if the spread was due to a specific clone.

Findings

Ten isolates carrying bla_CTX-M-1 from Swedish broilers belonged to eight different multi-locus sequence types with three isolates belonging to ST155. The ST155 isolates were identical as assessed by PFGE. The bla_CTX-M-1 was in all isolates carried on a plasmid of replicon type incI, which also transferred resistance to tetracycline and sulfamethoxazole.

Conclusion

The occurrence of ESBL-producing E. coli in the Swedish broilers is not due to the emergence of a single clone, but rather the spread of a specific incI plasmid carrying bla_CTX-M-1.     

The carry-over of Campylobacter isolates between sequential poultry flocks

The carry-over of Campylobacter strains from one flock to a subsequent flock in the same broiler house has been studied using molecular epidemiological techniques. In all, 524 Campylobacter strains, isolated from two sequential broiler flocks from 60 broiler houses, were typed by restriction fragment polymorphism of the polymerase chain reaction (PCR) product of the flaA and flaB genes (fla typing). Selected strains were also typed using pulsed field gel electrophoresis (PFGE). By fla typing, 15 (21%) of the 60 houses with Campylobacter-positive sequential flocks had identical genotypes. In 10 (16% overall) of these houses the strains were also identical by PFGE. The difference in PFGE patterns in the strains from the three remaining houses may be indicative of genetic instability. Overall, these results suggest that carry-over from one flock to a subsequent flock in the same house is a relatively infrequent event and, therefore, that routine broiler house cleansing and/or disinfection is largely adequate to eliminate Campylobacter contamination. An alternative explanation of the low level carry-over is a persistent source or reservoir, external to the environment of the broiler houses.     

Concentration and prevalence of Escherichia coli O157 and Salmonella serotypes in sheep during slaughter at two Australian abattoirs

Objective This study aimed to determine the presence and concentration of Escherichia coli O157 and Salmonella spp. on fleece, faeces and carcases of sheep during slaughter. Procedure Faeces, fleece and pre-chill carcase samples were collected from 164 sheep slaughtered at two Australian abattoirs. The presence of E. coli O157 and Salmonella spp. were determined by use of automated immunomagnetic separation (AIMS) with enumeration by use of the ‘most probable number’ (MPN) method. Results Escherichia coli O157 was isolated from 5% of faeces, 3% of fleeces and 0.6% of pre-chill carcases. The mean log₁₀ count of E. coli O157 positive faecal samples was 2.32 MPN/g, but counts on fleeces and carcases were below the countable limit (−1 log₁₀ MPN/cm²). Salmonella spp. were isolated from 20% of faeces, 13% of fleeces and 1.3% of pre-chill carcases. The mean log₁₀ count of Salmonella spp. in faeces was 1.43 MPN/g and on fleece was −0.24 MPN/cm², but counts on carcases were below the countable limit (−1 log₁₀ MPN/cm²). Conclusion The prevalence and concentration of pathogens were low in the sheep tested in this study, indicating a low risk of human infection from products derived from these animals.     

Systematic evaluation of in-house broiler litter windrowing effects on production benefits and environmental impact

In-house windrowing of broiler litter between flocks has been adopted by producers to reduce pathogens and improve litter quality before chick placement. In this study, 5 consecutive windrow trials were conducted in commercial broiler houses for their effect on litter bacterial populations, organic matter stabilization, cumulative ammonia emissions, and nutrient transformation and compared with litter conditioning (tilling) in adjacent houses. No significant reduction ofClostridium spp. andEscherichia coli populations was found in windrowed litter from d 0 to 7. No significant difference of 7-d mortality was found between windrow and nonwindrow houses. The windrowed house resulted in better foot quality than the nonwindrowed house from 1 of 3 scored flocks. Water-soluble phosphorus increased in both windrowed and nonwindrowed litter; therefore, appreciable biotic and abiotic activity occurred in litter with both treatments after flocks were removed. Overall, no negative effect of windrow treatments on litter quality for agronomic applications was observed. Both the control and windrow treatments resulted in a decrease in litter moisture content (2 to 5%) likely beneficial to bird health conditions. High ammonia emissions persisted after windrow spreading; therefore, a need may exist for an extended period of ventilation or a litter amendment as crucial before chick placement. Litter amendment at a low dose was effective in lowering ammonia concentrations after windrowing and was more economical comparing to operating fans in winter conditions.     

Integrated farm management to prevent Salmonella Enteritidis contamination of eggs

Salmonella Enteritidis in contaminated eggs is a public health hazard that may cause hospitalization or death in the elderly, infants, and individuals with impaired immune systems. Prevention of Salmonella Enteritidis infection of laying hens is an essential first step in reducing Salmonella Enteritidis outbreaks in humans. Multiple interventions at several stages during egg production can combine to reduce numbers of infected chickens and keep egg contamination to low levels. Every effort should be made to exclude Salmonella Enteritidis from egg production premises by implementing effective biosecurity measures, stocking the farm with Salmonella Enteritidis-free replacement pullets, controlling rodent and insect vectors, and denying wild birds and pets access to chicken houses. Diligent cleaning and disinfection of chicken houses before introduction of a new flock will minimize environmental exposure and indirect horizontal transmission of multiple pathogens, including Salmonella Enteritidis. Increased resistance of chickens to intestinal colonization by Salmonella Enteritidis can be attained by the use of probiotics, prebiotics, and synbiotics. Laying hens should be immunized with live and killed vaccines to stimulate mucosal and systemic immunity and reduce the prevalence of Salmonella Enteritidis-contaminated eggs. Shell eggs should be refrigerated as soon as possible after laying to keep Salmonella Enteritidis cells at low levels in any contaminated eggs. Comprehensive Salmonella Enteritidis-control programs have proven to be successful in reducing the incidence of Salmonella Enteritidis infections in both egg-laying flocks and humans.     

Airborne Gram‐Negative Bacterial Flora in Animal Houses

The concentration and the species composition of airborne gram‐negative bacteria were studied in four cattle houses, one pig house and one poultry barn. On average only between 0.02 and 5.2 % of the total number of culturable aerobic bacteria were identified as gram‐negative bacteria. Obligate anaerobic gram‐negative bacteria were not isolated at all. In the airborne gram‐negative bacterial flora the following bacterial families dominated: the Enterobacteriaceae, the Pseudomonadaceae and the Neisseriaceae. Within the family of the Enterobacteriaceae the species Escherichia coli and Enterobacter agglomerans were predominant. In animal houses using straw as bedding material Ent. agglomerans was most frequent, whereas in animal houses without litter E. coli was mainly found. Airborne Neisseriaceae were isolated very frequently in cow barns with Acinetobacter lowffii as the primary species. Airborne Pseudomonadaceae were found in high concentrations during periods of high air humidity. The results presented may also give some indications on the origin and sources of airborne endotoxins in animal housing.     

The effect of environmental poultry samples on the pH of typical Salmonella pre-enrichment and enrichment media following incubation

The first step to detect Salmonella in feed and other dry contaminated samples is a pre-enrichment broth that can assist in the recovery of small numbers of stressed Salmonella cells. A previous study demonstrated that incubation of feed and feed ingredients in commonly used pre-enrichment media resulted in a low pH that injured or killed the Salmonella. The objective of this study was to determine which environmental samples and pre-enrichment and selective broths could interfere with the accuracy of Salmonella detection by allowing pH to drop. Samples were collected from commercial poultry operations. Triplicate 10-g subunits were dispensed into sterile 18-oz Whirl Pak bags and 90 mL of each of the media [lactose broth (LB), buffered peptone water (BPW), Universal Pre-enrichment (UP), minimal salts (M-9), tetrathionate broth (TT) and Rappaport-Vassiliadis broth (RV)] were added to the bags and incubated 24 h at 37°C (or 42°C for RV and TT). The pH was measured after incubation. With turkey litter, fluff, and eggshells, regardless of media, the pH never went below 5.7. Regardless of sample type, the lowest pH was 6.1, 6.2, and 6.4 for UP, M-9, and BPW, respectively. For the pre-enrichment medium commonly used in the US, (LB) the pH dropped to 4.7 to 4.9 with broiler litter and 4.2 for boot covers used in turkey or broiler houses. Many researchers testing these sample types are unaware of this potential for pH change during pre-enrichment and may be underestimating the presence of Salmonella.