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Thermal resistance of desiccation-adapted Salmonella Senftenberg 775W was compared with that of Enterococcus faecium NRRL B-2354 in aged broiler litter. Aged broiler litter with 20, 30, and 40% moisture contents was inoculated separately with desiccation-adapted Salmonella Senftenberg 775W and E. faecium NRRL B-2354 at ca. 5 to 6 log CFU/g and then heat treated at 75, 85, and 150°C. At all tested temperatures, desiccation-adapted E. faecium NRRL B-2354 was more heat resistant than desiccation-adapted Salmonella Senftenberg 775W ( P < 0.05). During the treatments at 75 and 85°C, E. faecium NRRL B-2354 in aged broiler litter with all moisture contents was reduced by 2.89 to 4.12 log and was above the detection limit of direct plating (1.30 log CFU/g), whereas Salmonella Senftenberg 775W could not be detected by enrichment (>5-log reduction) during holding time at these temperatures. At 150°C, E. faecium NRRL B-2354 in aged broiler litter with 20 and 30% moisture contents was still detectable by enrichment after heat exposure for up to 15 min, whereas Salmonella Senftenberg 775W in aged broiler litter with all moisture contents could not be detected throughout the entire treatment. Our results revealed that E. faecium NRRL B-2354 can be used as a surrogate for Salmonella to validate the thermal processing of poultry litter by providing a sufficient safety margin. This study provides a practical tool for poultry litter processors to evaluate the effectiveness of their thermal processing.

This study evaluated the effectiveness of bacteriophage treatment for reducing Salmonella attachment and biofilms on hard surfaces. Bacteriophages (n = 6) were selected for bacteriophage treatment based on host ranges against Salmonella isolates (n = 10) obtained from rendering plants. The effectiveness of bacteriophage treatment (104-108 PFU/mL) was initially evaluated against strong Salmonella biofilm formers in 96-well microplate. Then, the bacteriophage treatment (109 PFU/mL) was applied for 7 d to reduce Salmonella attached to the stainless steel surfaces under laboratory and greenhouse conditions. The inhibition of biofilm formation and reduction of pre-formed biofilm in 96-well microplate with bacteriophage treatment reached up to 90 and 66%, respectively. Under laboratory conditions, bacteriophage treatment reduced up to 2.9 and 3.0 log CFU/cm2 of attachment and slightly formed biofilm of selected top 10 Salmonella strains and an avirulent Salmonella Typhimurium strain 8243, respectively, as compared with reductions of 3.4, 1.4, and 3.0 log CFU/cm2 of S. Typhimurium strain 8243 in summer, fall/winter, and spring seasons under greenhouse conditions, respectively. Clearly, bacteriophages were effective on reducing Salmonella attachment and biofilms formed on hard surfaces under both laboratory and greenhouse conditions. The use of bacteriophages on hard surfaces may have merits in reducing the likelihood of finished rendered products being recontaminated with Salmonella in rendering plants., (© 2017 Poultry Science Association Inc.)

A microbiological investigation on Salmonella contamination was conducted in two U.S. rendering plants to investigate the potential cross-contamination of Salmonella in the rendering processing environment. Sampling locations were predetermined at the areas where Salmonella contamination may potentially occur, including raw materials receiving, crax (rendered materials before grinding process) grinding, and finished meal loading-out areas. Salmonella was either enumerated directly on xylose lysine Tergitol 4 agar plates or enriched in Rappaport-Vassiliadis and tetrathionate broths. The presumptive Salmonella isolates were confirmed using CHROMagar plating and latex agglutination testing and then characterized using pulsed-field gel electrophoresis, serotyping, and biofilm-forming determination. Among 108 samples analyzed, 79 (73%) samples were Salmonella positive after enrichment. Selected Salmonella isolates (n = 65) were assigned to 31 unique pulsed-field gel electrophoresis patterns, with 16 Salmonella serotypes, including Typhimurium and Mbandaka, identified as predominant serotypes and 10 Salmonella strains determined as strong biofilm formers. Our results indicated that the raw materials receiving area was the primary source of Salmonella and that the surfaces surrounding crax grinding and finished meal loading-out areas harbor Salmonella in biofilms that may recontaminate the finished meals. The same Salmonella serotypes found in both raw materials receiving and the finished meal loading-out areas suggested a potential of cross-contamination between different areas in the rendering processing environment.

Broiler chicken litter was kept as a stacked heap on a poultry farm, and samples were collected up to 9 months of storage. Chicken litter inoculated with desiccation-adapted Salmonella cells was heat-treated at 75, 80, 85, and 150°C. Salmonella populations decreased in all these samples during heat treatment, and the inactivation rates became lower in chicken litter when storage time was extended from 0 to 6 months. There was no significant difference (P > 0.05) in thermal resistance of Salmonella in 6- and 9-month litter samples, indicating that a threshold for thermal resistance was reached after 6 months. Overall, the thermal resistance of Salmonella in chicken litter was affected by the storage time of the litter. The changes in some chemical, physical, and microbiological properties during storage could possibly contribute to this difference. Moisture and ammonia could be two of the most significant factors influencing the thermal resistance of Salmonella cells in chicken litter. Our results emphasize the importance of adjusting time and temperature conditions for heat processing chicken litter when it is removed from the chicken house at different time intervals., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

During the early phases of aerobic composting of animal manures, pathogens are inactivated primarily from the accumulation of heat produced by indigenous microbial activity. When compost materials are not exposed to these lethal temperatures, the required holding time needed to obtain a pathogen-free product that may be applied to fields is unknown. Consequently, a series of studies examined whether the carbon amendment (wheat straw, peanut hulls, rice hulls, and pine needles) added to animal manures affected survival of either Salmonella or E. coli O157:H7 during storage of compost mixtures at sublethal temperatures (20 to 40°C). Pathogens consistently survived for longer periods of time in compost mixtures prepared with pine needles than compost mixtures prepared with either of the other three carbon amendments. Pathogen inactivation in wheat straw- or peanut hull-amended compost mixtures was dependent on the target pathogen, moisture level, and storage temperature. Moisture levels in wheat straw-amended compost mixtures stored at 40°C had no effect on inactivation of E. coli O157:H7. In contrast, wheat straw-amended mixtures stored at 30 to 35°C and equilibrated to suboptimal moisture contents (30 to 40%) were less effective for inactivating pathogens compared with drier (25% moisture) or moister (60% moisture) mixtures. In peanut hull-amended compost mixtures, inactivation of E. coli O157:H7 was affected minimally by moisture levels, whereas Salmonella survival increased as the moisture level was decreased. The different inactivation responses of Salmonella and E. coli O157:H7 in compost mixtures prepared with wheat straw or peanut hulls and equilibrated to different moisture levels suggest that there are different mechanisms for inactivation. Hence, developing reliable guidelines relying on time-temperature for holding of compost mixtures at sublethal temperatures will be challenging and, perhaps, not possible.

The effectiveness of a two-step heat treatment for eliminating desiccation-adapted Salmonella spp. in aged chicken litter was evaluated. The aged chicken litter with 20, 30, 40, and 50% moisture contents was inoculated with a mixture of four Salmonella serotypes for a 24-h adaptation. Afterwards, the inoculated chicken litter was added into the chicken litter with the adjusted moisture content for a 1-h moist-heat treatment at 65 °C and 100% relative humidity inside a water bath, followed by a dry-heat treatment in a convection oven at 85 °C for 1 h to the desired moisture level (<10-12%). After moist-heat treatment, the populations of Salmonella in aged chicken litter at 20 and 30% moisture contents declined from ≈6.70 log colony-forming units (CFU)/g to 3.31 and 3.00 log CFU/g, respectively. After subsequent 1-h dry-heat treatment, the populations further decreased to 2.97 and 2.57 log CFU/g, respectively. Salmonella cells in chicken litter with 40% and 50% moisture contents were only detectable by enrichment after 40 and 20 min of moist-heat treatment, respectively. Moisture contents in all samples were reduced to <10% after a 1-h dry-heat process. Our results demonstrated that the two-step heat treatment was effective in reducing >5.5 logs of desiccation-adapted Salmonella in aged chicken litter with moisture content at or above 40%. Clearly, the findings from this study may provide the chicken litter processing industry with an effective heat treatment method for producing Salmonella-free chicken litter.

Two separate studies were conducted to address the condition and the type of feedstocks used during composting of dairy manure. In each study, physical (temperature), chemical (ammonia, volatile acids, and pH), and biological (Salmonella, Listeria monocytogenes, and Escherichia coli O157:H7) parameters were monitored during composting in bioreactors to assess the degree to which they were affected by the experimental variables and, ultimately, the ability of the chemical and physical parameters to predict the fate of pathogens during composting. Compost mixtures that contained either aged dairy manure or pine needles had reduced heat generation; therefore, pathogen reduction took longer than if fresh manure or carbon amendments of wheat straw or peanut hulls were used. Based on regression models derived from these results, ammonia concentration, in addition to heat, were the primary factors affecting the degree of pathogen inactivation in compost mixtures formulated to an initial carbon-nitrogen (C:N) ratio of 40:1, whereas, the pH of the compost mixture along with the amount of heat exposure were most influential in compost mixtures formulated to an initial C:N ratio of 30:1. Further studies are needed to validate these models so that additional criteria in addition to time and temperature can be used to evaluate the microbiological safety of composted manures.

Thermal inactivation of desiccation-adapted Salmonella spp. in aged chicken litter was investigated in comparison with that in a nonadapted control to examine potential cross-tolerance of desiccation-adapted cells to heat treatment. A mixture of four Salmonella serovars was inoculated into the finished compost with 20, 30, 40, and 50% moisture contents for a 24-h desiccation adaptation. Afterwards, the compost with desiccation-adapted cells was inoculated into the aged chicken litter with the same moisture content for heat treatments at 70, 75, 80, 85, and 150°C. Recovery media were used to allow heat-injured cells to resuscitate. A 5-log reduction in the number of the desiccation-adapted cells in aged chicken litter with a 20% moisture content required >6, >6, ∼4 to 5, and ∼3 to 4 h of exposure at 70, 75, 80, and 85°C, respectively. As a comparison, a 5-log reduction in the number of nonadapted control cells in the same chicken litter was achieved within ∼1.5 to 2, ∼1 to 1.5, ∼0.5 to 1, and <0.5 h at 70, 75, 80, and 85°C, respectively. The exposure time required to obtain a 5-log reduction in the number of desiccation-adapted cells gradually became shorter as temperature and moisture content were increased. At 150°C, desiccation-adapted Salmonella cells survived for 50 min in chicken litter with a 20% moisture content, whereas control cells were detectable by enrichment for only 10 min. Our results demonstrated that the thermal resistance of Salmonella in aged chicken litter was increased significantly when the cells were adapted to desiccation. This study also validated the effectiveness of thermal processing being used for producing chicken litter free of Salmonella contamination.

This study analyzed various organic fertilizers for indicator microorganisms, pathogens, and antibiotic-resistant Escherichia coli, and evaluated the growth potential of E. coli O157:H7 and Salmonella in fertilizers. A microbiological survey was conducted on 103 organic fertilizers from across the United States. Moisture content ranged from approximately 1% to 86.4%, and the average pH was 7.77. The total aerobic mesophiles ranged from approximately 3 to 9 log colony-forming units (CFU)/g. Enterobacteriaceae populations were in the range of <1 to approximately 7 log CFU/g, while coliform levels varied from <1 to approximately 6 log CFU/g. Thirty samples (29%) were positive for E. coli, with levels reaching approximately 6 log CFU/g. There were no confirmed positives for E. coli O157:H7, Salmonella, or Listeria monocytogenes. The majority of E. coli isolates (n=73), confirmed by glutamate decarboxylase (gad) PCR, were from group B1 (48%) and group A (32%). Resistance to 16 antibiotics was examined for 73 E. coli isolates, with 11 isolates having resistance to at least one antibiotic, 5 isolates to ≥ 2 antibiotics, and 2 isolates to ≥ 10 antibiotics. In the presence of high levels of background aerobic mesophiles, Salmonella and E. coli O157:H7 grew approximately 1 log CFU/g within 1 day of incubation in plant-based compost and fish emulsion-based compost, respectively. With low levels of background aerobic mesophiles, Salmonella grew approximately 2.6, 3.0, 3.0, and 3.2 log CFU/g in blood, bone, and feather meals and the mixed-source fertilizer, respectively, whereas E. coli O157:H7 grew approximately 4.6, 4.0, 4.0, and 4.8 log CFU/g, respectively. Our results revealed that the microbiological quality of organic fertilizers varies greatly, with some fertilizers containing antibiotic resistant E. coli and a few supporting the growth of foodborne pathogens after reintroduction into the fertilizer.

Our results revealed that a 7-log reduction of Salmonella can be achieved by exposing fresh chicken litter for 80.5 to 100.8, 78.4 to 93.1, and 44.1 to 63 min at 70, 75, and 80°C, respectively, depending on initial moisture contents. However, the aged chicken litter requires more heat treatment.

Thermal resistance of heat-shocked Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes was compared with that of non-heat-shocked (control) strains in finished dairy compost. A three-strain mixture of each pathogen was heat shocked at 47.5°C for 1 h and inoculated into the compost at a final concentration of 10(7) CFU/g. The inoculated compost was placed inside an environmental chamber set at 50, 55, or 60°C with humidity at ca. 70%. The heat-shocked E. coli O157:H7, Salmonella, and L. monocytogenes survived better (P < 0.05) at 50°C, with reductions of 2.7, 3.2, and 3.9 log CFU/g within 4 h compared with reductions of 3.6, 4.5, and 5.1 log CFU/g, respectively, in control cultures. The heat-shocked cultures of E. coli O157:H7, Salmonella, and L. monocytogenes had 1.2-, 1.9-, and 2.3-log reductions, respectively, within 1 h at 55°C, whereas the corresponding control cultures had 4-, 5.6-, and 4.8-log reductions, respectively. At 60°C, a rapid population reduction was observed during the come-up time of 14 min in control cultures of E. coli O157:H7, Salmonella, and L. monocytogenes with 4.9-, 4.8-, and 2.3-log reductions, respectively, compared with 2.6-, 2.4-, 1.7-log reductions, respectively, in heat-shocked cultures. L. monocytogenes survival curves for all three temperatures had extensive tailing. The double Weibull distribution model was a good fit for the survival curves of pathogens, with differences in the shape parameter of heat-shocked and control cultures. Our results suggest that the heat-shocked pathogens may have extended survival at lethal temperatures attained during the composting process.

The objectives of this study were to determine the current status of bacterial contamination in rendered animal products and to analyze Salmonella and enterococci isolates from the samples. One hundred and fifty samples were provided by various rendering companies across the United States, including the following meal types: feather, meat, meat and bone, meat and bone from poultry, poultry, and blood meals. The average pH of the meals ranged from 6.16 to 7.36, and the moisture content ranged from 1.9% to 11.5%. The total bacterial counts were in the range of 1.7 to 6.68 log10 CFU/g, with the highest in blood meal and the lowest in meat meal. Enterococcus species were detected in 81.3% of the samples and accounted for up to 54% of the total bacterial counts in some samples. Both blood meal and feather meal were more contaminated (P < 0.05) with enterococci than other meal types, although all blood meals were from a single company. Salmonella was detected in 8.7% of the samples. Escherichia coli was not detected in any of the samples, but coliforms were detected in four samples. Among enterococci isolates, three were vancomycin resistant. Thirteen serotypes of Salmonella displayed 16 pulsed-field gel electrophoresis patterns. Pulsed-field gel electrophoresis analysis has indicated that Salmonella contamination was not persistent in the plant environment over time. The D-values for the Salmonella isolates at 55, 60, and 65 degrees C were in the ranges of 9.27-9.99, 2.07-2.28, and 0.35-0.40 min, respectively. These results suggest that the presence of Salmonella in the finished products may be due to postprocessing contamination. This study has also revealed that the rendering industry has microbiologically improved its products since earlier studies were conducted.

Aerobic composting is a common management practice to inactivate pathogens in manure; however, additional research on the role of compost composition in pathogen inactivation is needed. The objective of this study was therefore to determine the effect of the carbon:nitrogen (C:N) ratio and the presence of ammonium sulfate on inactivation of Salmonella spp. in cow manure-based mixtures composted in a bioreactor under controlled conditions. Compost preparations with an initial C:N ratio of 20:1 required a maximum of 4 days of storage before Salmonellae were inactivated by 7 log(10), whereas preparations with C:N ratios of 30:1 and 40:1 C:N required more than 5 and 7 days of storage, respectively. The pH values of both the 20:1 and 30:1 C:N preparations decreased during the onset of composting before increasing to >8. In contrast, pH values of 40:1 C:N preparations increased immediately to >8, generally within the first day of storage. Maximum temperatures observed in 20:1 C:N preparations for inactivation of pathogens were less than 50 degrees C, and the cumulative heat exposure required for pathogen inactivation in 20:1 C:N preparations was 15-fold less than in 40:1 C:N preparations. Supplementation of compost mixtures with 0.08% ammonium sulfate resulted in slightly higher temperatures; however, these higher temperatures did not translate into more rapid rates of pathogen inactivation.

Pathogenic microorganisms may survive the composting process in low numbers and subsequently regrow to high levels under favorable conditions. The objective of this study was to investigate the regrowth potential of Salmonella spp., Escherichia coli O157:H7, and Listeria monocytogenes in dairy-based composts under different environmental conditions. Water extract of commercially available dairy compost was used as a model system. Cocktails of five rifampin-resistant strains of each pathogen previously grown in reduced nutrient media (1/2 or 1/10 strength of tryptic soy broth, TSB) were inoculated into water extract of compost of different ratios (1:2,1:5, and 1:10, w/v), and then stored at 35 degrees C or 22 degrees C for 7 days. The strains exhibiting greatest survival or regrowth were identified by pulsed-field gel electrophoresis (PFGE). At 22 degrees C, both E. coli O157:H7 and L. monocytogenes multiplied in all compost extracts, whereas Salmonella spp. regrew in both 1:2 and 1:5 compost extracts but not in 1:10. For all three pathogens, incubation at 22 degrees C provides better conditions for regrowth than at 35 degrees C. Both Salmonella and E. coli O157:H7 previously adapted to nutrient-limited broth (1/10 strength of TSB) regrew in compost extracts to higher populations than the control cultures grown previously in full strength of TSB. In the absence of indigenous microorganisms, all three pathogens regrew even in the most diluted sterile compost extract (1:10) with growth potentials ranging from 2.30 to 3.59 log CFU/ml. In nonsterile compost extract with ca. 5 log CFU/ml of background microorganisms, all three pathogens regrew only in the most concentrated compost extract (1:2) with much less population increases ranging from 0.70 to 1.43 log CFU/ml. Compost extract samples of all ages supported the regrowth of both Salmonella and E. coli O157:H7 with population increases ranging from 0.95 to 2.32 log CFU/ml. The PFGE patterns for E. coli O157:H7 isolates from sterile compost extracts matched with either the spinach outbreak strain or an avirulent B6914 strain. These results demonstrated that compost extract of dairy-based compost contained sufficient nutrients for pathogen regrowth. Cultures previously adapted to low nutrient media regrew to higher populations than control cultures; however, indigenous microflora suppressed the pathogen regrowth in compost extract, especially at 35 degrees C.

The objective of this study was to assess and differentiate wild-caught South Carolina (SC) shrimps from imported shrimps on the basis of microbiological analysis. Seven wild-caught SC shrimp and 13 farm-raised imported shrimp samples were analyzed. Total plate counts from wild-caught shrimp samples ranged from 4.3 to 7.0 log10 CFU/g, whereas counts from imported shrimp samples ranged from 3.2 to 5.7 log10 CFU/g. There was no difference (P > 0.05) between total bacterial counts of wild-caught SC shrimp and farm-raised imported shrimp. However, the percentages of bacteria with reduced susceptibility towards ceftriaxone and tetracycline were higher (P < 0.05) for farm-raised shrimp than for wild-caught samples. Salmonella spp. detected only in one farm-raised sample was resistant to ampicillin, ceftriaxone, gentamicin, streptomycin, and trimethoprim. Vibrio vulnificus was detected in both wild-caught and farm-raised shrimp samples; however, only the isolate from farm-raised shrimp was resistant to nalidixic acid and trimethoprim. Escherichia coli detected in one wild-caught sample was resistant to ampicillin. Both Listeria spp. and Salmonella spp. were absent with wild-caught SC samples. Therefore, the presence of more ceftriaxone- and tetracycline-resistant bacteria and the observed antimicrobial resistance phenotypes of isolates from the imported shrimp may reflect the possible use of antibiotics in raising shrimp in those countries.

Compost is organic material that has been degraded into a nutrient-stabilized humus-like substance through intense microbial activity, which can provide essential plant nutrients (nitrogen, phosphorus) to aid in the growth of fruits and vegetables. Compost can be generated from animal waste feedstocks; these can contain human pathogens, which can be inactivated through the heat and microbial competition promoted during the composting process. Outbreaks of infections caused by bacterial pathogens such as Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes on fruit and vegetable commodities consumed raw emphasize the importance of minimizing the risk of pathogenic contamination on produce commodities. This review article investigates factors that affect the reduction and survival of bacterial foodborne pathogens during the composting process. Interactions with indigenous microorganisms, carbon:nitrogen ratios, and temperature changes influence pathogen survival, growth, and persistence in finished compost. Understanding the mechanisms of pathogen survival during the composting process and mechanisms that reduce pathogen populations can minimize the risk of pathogen contamination in the cultivation of fruits and vegetables. [ABSTRACT FROM AUTHOR]

Abstract: This study investigated the survival of heat-shocked (HS) and non-heat-shocked (NHS) Escherichia coli O157:H7 and Salmonella enterica Typhimurium when co-composting dairy manure and vegetable wastes in a field setting. In the summer, HS E. coli O157:H7 and Salmonella survived for 7 and 2days longer at the surface and bottom locations of the compost heaps, respectively, than NHS cultures. Both HS and NHS E. coli O157:H7 and Salmonella were detectable in all compost samples for more than 60days in the winter. The results indicate that composting dairy manure with vegetable wastes under sub-optimal conditions may allow extended survival of pathogens in the heap at low ambient temperature. Analysis of covariance revealed that the heat-shock treatment may have induced cross-resistance to desiccation, allowing extended survival of HS E. coli O157:H7 and Salmonella at the surface of the compost heaps during the summer. [Copyright &y& Elsevier]

Abstract: The fate of manure-borne pathogen surrogates (gfp-labeled Escherichia coli O157:H7 and Listeria innocua and avirulent Salmonella Typhimurium) in the field was monitored at both sub-surface (30cm from surface) and surface sites of static composting piles (3.5-m base diameter) composed of chicken litter and peanut hulls. Despite exposure to elevated temperatures, Salmonella was detected by enrichment culture in sub-surface samples following 14days of composting. In surface samples, pathogen surrogates were detected in the summer after 4days of composting by enrichment culture only, whereas E. coli O157:H7 and L. innocua remained detectable by direct plating (>2log10 cfu/g) up to 28days in piles composted during the fall and winter. All three types of bacteria remained detectable by enrichment culture in surface samples composted for 56days during the winter. [Copyright &y& Elsevier]