Cardiac Magnetic Resonance Studies in a Large Animal Model That Simulates the Cardiac Abnormalities of Human Septic Shock.

BACKGROUND: Septic shock is associated with increases in end-diastolic volume (EDV) and decreases in ejection fraction that reverse within 10 days. Nonsurvivors do not develop EDV increases. The mechanism is unknown. METHODS AND RESULTS: Purpose-bred beagles (n=33) were randomized to receive intrabronchial Staphylococcus aureus or saline. Over 96 hours, cardiac magnetic resonance imaging and echocardiograms were performed. Tissue was obtained at 66 hours. From 0 to 96 hours after bacterial challenge, septic animals versus controls had significantly increased left ventricular wall edema (6%) and wall thinning with loss of mass (15%). On histology, the major finding was nonocclusive microvascular injury with edema in myocytes, the interstitium, and endothelial cells. Edema was associated with significant worsening of biventricular ejection fractions, ventricular-arterial coupling, and circumferential strain. Early during sepsis, (0-24 hours), the EDV decreased; significantly more in nonsurvivors (ie, greater diastolic dysfunction). From 24 to 48 hours, septic animals' biventricular chamber sizes increased; in survivors significantly greater than baseline and nonsurvivors, whose EDVs were not different from baseline. Preload, afterload, or heart rate differences did not explain these differential changes. CONCLUSIONS: The cardiac dysfunction of sepsis is associated with wall edema. In nonsurvivors, at 0 to 24 hours, sepsis induces a more severe diastolic dysfunction, further decreasing chamber size. The loss of left ventricular mass with wall thinning in septic survivors may, in part, explain the EDV increases from 24 to 48 hours because of a potentially reparative process removing damaged wall tissue. Septic cardiomyopathy is most consistent with a nonocclusive microvascular injury resulting in edema causing reversible systolic and diastolic dysfunction with more severe diastolic dysfunction being associated with a decreased EDV and death.

Cardiac Magnetic Resonance Studies in a Large Animal Model that Simulates the Cardiac Abnormalities of Human Septic Shock.

Background: Septic shock, in humans and in our well-established animal model, is associated with increases in biventricular end diastolic volume (EDV) and decreases in ejection fraction (EF). These abnormalities occur over 2 days and reverse within 10 days. Septic non-survivors do not develop an increase in EDV. The mechanism for this cardiac dysfunction and EDV differences is unknown. Methods: Purpose-bred beagles randomized to receive intrabronchial Staphylococcus aureus (n=27) or saline (n=6) were provided standard ICU care including sedation, mechanical ventilation, and fluid resuscitation to a pulmonary arterial occlusion pressure of over 10mmHg. No catecholamines were administered. Over 96h, cardiac magnetic resonance imaging, echocardiograms, and invasive hemodynamics were serially performed, and laboratory data was collected. Tissue was obtained at 66h from six septic animals. Results: From 0-96h after bacterial challenge, septic animals vs. controls had significantly increased left ventricular wall edema (6%) and wall thinning with loss of mass (15%) which was more pronounced at 48h in non-survivors than survivors. On histology, edema was located predominantly in myocytes, the interstitium, and endothelial cells. Edema was associated with significantly worse biventricular function (lower EFs), ventricular-arterial coupling, and circumferential strain. In septic animals, from 0-24h, the EDV decreased from baseline and, despite cardiac filling pressures being similar, decreased significantly more in non-survivors. From 24-48h, all septic animals had increases in biventricular chamber sizes. Survivors biventricular EDVs were significantly greater than baseline and in non-survivors, where biventricular EDVs were not different from baseline. Preload, afterload, or HR differences did not explain these differential serial changes in chamber size. Conclusion: Systolic and diastolic cardiac dysfunction during sepsis is associated with ventricular wall edema. Rather than differences in preload, afterload, or heart rate, structural alterations to the ventricular wall best account for the volume changes associated with outcome during sepsis. In non-survivors, from 0-24h, sepsis induces a more severe diastolic dysfunction, further decreasing chamber size. The loss of left ventricular mass with wall thinning in septic survivors may, in part explain, the EDV increases from 24-48h. However, these changes continued and even accelerated into the recovery phase consistent with a reparative process rather than ongoing injury.

Management Practices Affecting Lesser Mealworm Larvae (Alphitobius diaperinus) Associated Microbial Community in a Broiler House and After Relocating With the Litter Into Pastureland.

Lesser mealworms are often found infesting production houses used to raise broiler chickens. Previous studies have investigated pathogenic microorganisms associated with the larvae, but a more thorough study relating total microbiome changes due to management procedures and flock rotations was needed. Additionally, there is a question of what microbiota are transferred into the environment when the litter, in which larvae reside, is piled in pastureland for use as fertilizer and where interactions with the soil and other fauna can occur. This study chronicled, by the 16S rRNA sequencing, the bacterial community profile of larvae in a broiler grow-out house synchronizing to when birds were added to and removed from the house over 2.5 years. The profile was found to be relatively constant despite 11 flock rotations and management disruptions, specifically litter cleanout procedures and the addition of new birds or bedding. In contrast, once removed from the controlled broiler house environment and placed into open pastureland, the substantial microbial diversity brought with the larvae showed greater fluctuation in structure with environmental conditions, one of which was rainfall. Surprisingly, these larvae survived at least 19 weeks, so the potential for moving larval-associated microbes into the environment needs further assessment to minimize the risk of relocating foodborne pathogens and also to assess those bacteria-generating metabolites that have benefits to plant growth when using the litter as a fertilizer. The characterization of their microbiome is the first step to investigating the influences of their microbes on the manmade and environmental ecosystems.

Adult Alphitobius diaperinus Microbial Community during Broiler Production and in Spent Litter after Stockpiling.

The facilities used to raise broiler chickens are often infested with litter beetles (lesser mealworm, Alphitobius diaperinus). These beetles have been studied for their carriage of pathogenic microbes; however, a more comprehensive microbiome study on these arthropods is lacking. This study investigated their microbial community in a longitudinal study throughout 2.5 years of poultry production and after the spent litter, containing the mealworms, was piled in pastureland for use as fertilizer. The mean most abundant phyla harbored by the beetles in house were the Proteobacteria (39.8%), then Firmicutes (30.8%), Actinobacteria (21.1%), Tenericutes (5.1%), and Bacteroidetes (1.6%). The community showed a modest decrease in Firmicutes and increase in Proteobacteria over successive flock rotations. The beetles were relocated within the spent litter to pastureland, where they were found at least 19 weeks later. Over time in the pastureland, their microbial profile underwent a large decrease in the percent of Firmicutes (20.5%). The lesser mealworm showed an ability to survive long-term in the open environment within the spent litter, where their microbiome should be further assessed to both reduce the risk of transferring harmful bacteria, as well as to enhance their contribution when the litter is used as a fertilizer.

Characterization of Salmonella enterica isolates from turkeys in commercial processing plants for resistance to antibiotics, disinfectants, and a growth promoter.

Salmonella enterica isolates from turkeys in two commercial processing plants (1 and 2) were characterized for susceptibility to antibiotics, disinfectants, and the organoarsenical growth promoter, 4-hydroxy-3-nitrophenylarsonic acid (3-NHPAA, roxarsone), and it's metabolites, NaAsO(2) (As(III)) and Na(2)HAsO(4) • 7H(2)O (As(V)). The 130 Salmonella serovars tested demonstrated a low incidence of resistance to the antibiotics gentamicin (GEN), kanamycin (KAN), sulfamethoxazole (SMX), streptomycin (STR), and tetracycline (TET). Isolates resistant to antibiotics were most often multidrug resistant. Serovars Hadar and Typhimurium were resistant to KAN, STR, and TET and GEN, SMX, and STR, respectively. All isolated Salmonella serovars were resistant to the disinfectant chlorhexidine with minimum inhibitory concentrations (MICs; 1-8 µg/mL), and they were susceptible to triclosan and benzalkonium chloride. The didecyldimethylammonium chloride component was the most active ammonium chloride tested. No cross-resistance was observed between antibiotics and disinfectants. The MICs for 3-NHPAA (4096 µg/mL) were consistent between processing Plant 1 and Plant 2, but MICs for the 3-NHPAA metabolites (As(III) and As(V)) were higher in Plant 1 than in Plant 2. In Plant 1, 76% of the isolates had MICs >256 µg/mL for As(III) and 92% of the isolates had MICs >1024 µg/mL for As(V). In Plant 2, all of the isolates had MICs ≤256 µg/mL for As(III) and 90% of the isolates had MICs ≤1024 µg/mL for As(V). Only 4 Salmonella serovars were isolated from Plant 1, but 10 serovars were isolated from Plant 2. S. enterica serovar Derby from Plant 1 was highly resistant to As(III) and As(V) with MICs >1024 and >8192 µg/mL, respectively, suggesting previous exposure to high arsenic metabolite concentrations. These levels may have been high enough to kill other Salmonella serovars, thus possibly explaining the lack of serovar diversity observed in Plant 1. The application of a growth promoter may affect the serovar diversity in treated birds.

Poultry litter and the environment: Microbial profile of litter during successive flock rotations and after spreading on pastureland.

With the increasing demand for broiler meat, a thorough evaluation of the microbial community within the broiler houses and sites where litter is deposited is critical to animal and environmental wellbeing. However not much is known in this arena, so our work evaluates the litter bacterial microbial community within a house over a 2.5 year period through 11 flock rotations, a partial and a total cleanout, and the subsequent deposition of the litter as fertilizer on pastureland. The effects of both time and management practices correlated with alterations of the litter microbial community. The cleanout practices and introduction of new bedding had minimal influence on the house microbial community once it was established, which generally showed a consistent increase in the proportion of Actinobacteria and a decrease in Firmicutes over the 11 flock rotations. Analysis of the bacterial profile at the genus level gave increased resolution, revealing changes during the first and second flock rotation and after the total cleanout. The disturbance of the partial cleanout seemed to be buffered by the supporting conditions within the house while the total cleanout showed a small, but significant influence. The pastureland deposition of litter, however, was affected by time and abiotic factors that changed the litter microbial community structure weekly. The stockpiled litter had an increase in the phyla Actinobacteria and the class Bacilli that commonly have microbes utilizing nitrogen and decaying materials, in comparison to Native soil. Further, the soil beneath where the litter was stored for 20 weeks, lost diversity, indicating a possible effect of the litter stockpiling on environmental quality at that site. How management practices affect the composition of the microbial community within the litter of the broiler house is of interest in terms of bird health and environmentally for future utilization of spent litter.

Characterization of planktonic and biofilm communities of day-of-hatch chicks cecal microflora and their resistance to Salmonella colonization.

Recent concerns about the use of antimicrobials in food animals have increased interest in the microbial ecology and biofilms within their gastrointestinal tract. This work used a continuous-flow chemostat system to model the microbial community within the ceca from day-of-hatch chicks and its ability to resist colonization by Salmonella enterica serovar Typhimurium. We characterized the biofilm and planktonic communities from five cultures by using automated ribotyping. Eight species from six different genera were identified. Overall, the planktonic communities were more diverse, with 40% of the cultures containing four or more bacterial species. Eighty percent of the biofilm communities contained only one or two species of bacteria. Enterococcus faecalis was the only species isolated from all communities. None of the resulting microbial communities was able to resist colonization by S. enterica serovar Typhimurium. This is the first study to provide a molecular-based characterization of the biofilm and planktonic communities found in day-of-hatch chicken cecal microflora cultures.

How Management Practices Within a Poultry House During Successive Flock Rotations Change the Structure of the Soil Microbiome.

The microbiome within a poultry production house influences the attainment of physiologically strong birds and thus food safety and public health. Yet little is known about the microbial communities within the house and the effects on the soil microbes onto which the houses are placed; nor the effects of management practices on their equilibrium. This study looked at the soil bacterial microbiome before a broiler house was constructed, then through 11 flock rotations (2.5 years) that included a partial clean-out and a total clean-out within the management regimen. Major shifts were observed, occurring at the taxonomic class level, related to the introduction of bedding and birds on top of the soil. The partial clean-out of litter did not change the soil bacterial community in any substantial way, only prompting a temporary increase in some genera; however, the total litter clean-out caused a major increase in a cohort of Actinobacteria. The underlying soil contained bacteria beneficial for poultry metabolism, such as Lactobacillus, Faecalibacterium, Bacteriodes, and Ruminococcus. Additionally, management practices affected the class structure of the soil bacterial community beneath the poultry house. The scheduling of these practices should be leveraged to exploit maintenance of beneficial bacteria that maximize microbiome contributions to bird production processes, while minimizing possible antibiotic-resistant bacteria and environmental effects.

Planktonic and biofilm community characterization and Salmonella resistance of 14-day-old chicken cecal Microflora-derived continuous-flow cultures.

This study evaluated the composition of gastrointestinal bacterial communities in birds during an age in which their susceptibility to Salmonella is highly diminished. One of the challenges to developing probiotics is to develop an efficacious culture of minimal diversity that includes bacteria that are vital contributors to protection from pathogens, but excludes unnecessary species. This study used in vitro continuous-flow culture techniques to test the ability of mixed bacterial cultures acquired from in vivo sources, to resist colonization by a marker Salmonella enterica serovar Typhimurium, and then characterized the constituents of both biofilm and planktonic communities by biochemical, phenotypic, and molecular methods. These cultures, initiated from 14-day-old chicks, were all able to restrict colonization by Salmonella in an average of 10 days. Eighteen species of bacteria from 10 different genera were characterized. However, each culture contained a mixture of only 11 species, which included lactic acid bacteria. Biofilms contained less than 50% of the species found in the planktonic communities. Although not adults, the diversity of microbes within the cecal cultures from 14-day-old birds represents a community complex enough to oppose colonization by a nonindigenous bacteria in vitro. These results describe bacterial mixtures containing less diversity than in previously described avian protective cultures.

External surface disinfection of the lesser mealworm (Coleoptera: Tenebrionidae).

Understanding the dynamics of movement of bacteria within the environment and between species is crucial to unraveling the epidemiology of bacterial diseases and to developing biosecurity measures to prevent dissemination. Many arthropods, some beneficial and some detrimental, inhabit poultry houses. The lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), is a pest commonly found in poultry litter that can harbor pathogens involved in both human and animal health issues. Current farm management practices perpetuate persistent infestations contributing to the dispersal of beetles and pathogens. To study the dissemination of bacteria by this beetle, we require the ability to differentiate internal from external sources of bacteria carried by the beetle. In this study, we tested previously described methods to externally disinfect beetles and found disinfectant efficacies between 40 and 98%. The irregular surface of the insect posed a challenge to cleansing procedures because the surface offered many recesses able to sequester bacteria. Complete bacterial disinfection was achieved with a serial treatment of ethanol and hydrogen peroxide or hydrogen peroxide/peracetic acid.

Dereplication by automated ribotyping of a competitive exclusion culture bacterial isolate library.

Concerns over the development of antibiotic-resistant bacteria within the food animal industry have intensified the search for natural approaches to the prevention and treatment of bacterial diseases. Competitive exclusion cultures are the foundation of a disease-management strategy based on the use of benign bacterial strains to prevent the establishment of pathogenic bacteria within a specific host. Differentiation of phenotypically ambiguous isolates is a critical step in establishing a manageable library of bacteria for use in the development of defined competitive exclusion cultures. We used automated ribotyping techniques to dereplicate a large collection of phenotypically ambiguous isolates from a continuous-flow competitive exclusion culture. A total of 157 isolates were screened following an EcoRI restriction enzyme digestion. The 157 isolates were resolved into 23 ribogroups, which represents an 85% reduction in the number of isolates in the bacterial isolate library. Seventy-six percent of the isolates fit into one of five ribogroups. This work demonstrated that automated ribotyping is an effective and efficient tool for dereplication of diverse bacterial isolate libraries.

Poultry litter and the environment: Physiochemical properties of litter and soil during successive flock rotations and after remote site deposition.

The U.S. broiler meat market has grown over the past 16 years and destinations for U.S. broiler meat exports expanded to over 150 countries. This market opportunity has spurred a corresponding increase in industrialized poultry production, which due to the confined space in which high numbers of animals are housed, risks accumulating nutrients and pollutants. The purpose of this research was to determine the level of pollutants within poultry litter and the underlying soil within a production facility; and to explore the impact of spent litter deposition into the environment. The study follows a production facility for the first 2.5 years of production. It monitors the effects of successive flocks and management practices on 15 physiochemical parameters: Ca, Cu, electrical conductivity, Fe, K, Mg, Mn, moisture, Na, NO3(-)/N, organic matter, P, pH, S, and Zn. Litter samples were collected in-house, after clean-outs and during stockpiling. The soil before house placement, after the clean-outs and following litter stockpiling was monitored. Management practices markedly altered the physiochemical profiles of the litter in-house. A canonical discriminant analysis was used to describe the relationship between the parameters and sampling times. The litter profiles grouped into five clusters corresponding to time and management practices. The soil in-house exhibited mean increases in all physiochemical parameters (2-297 fold) except Fe, Mg, %M, and pH. The spent litter was followed after deposition onto a field for use as fertilizer. After 20 weeks, the soil beneath the litter exhibited increases in EC, Cu, K, Na, NO3(-)/N, %OM, P, S and Zn; while %M decreased. Understanding the impacts of industrialized poultry farms on the environment is vital as the cumulative ecological impact of this land usage could be substantial if not properly managed to reduce the risk of potential pollutant infiltration into the environment.

Disinfectant and Antimicrobial Susceptibility Profiles of the Big Six Non-O157 Shiga Toxin-Producing Escherichia coli Strains from Food Animals and Humans.

The disinfectant and antimicrobial susceptibility profiles of 138 non-O157 Shiga toxin-producing Escherichia coli strains (STECs) from food animals and humans were determined. Antimicrobial resistance (AMR) was moderate (39.1% of strains) in response to 15 antimicrobial agents. Animal strains had a lower AMR prevalence (35.6%) than did human strains (43.9%) but a higher prevalence of the resistance profile GEN-KAN-TET. A decreasing prevalence of AMR was found among animal strains from serogroups O45 > O145 > O121 > O111 > O26 > O103 and among human strains from serogroups O145 > O103 > O26 > O111 > O121 > O45. One animal strain from serogroups O121 and O145 and one human strain from serogroup O26 had extensive drug resistance. A high prevalence of AMR in animal O45 and O121 strains and no resistance or a low prevalence of resistance in human strains from these serogroups suggests a source other than food animals for human exposure to these strains. Among the 24 disinfectants evaluated, all strains were susceptible to triclosan. Animal strains had a higher prevalence of resistance to chlorhexidine than did human strains. Both animal and human strains had a similar low prevalence of low-level benzalkonium chloride resistance, and animal and human strains had similar susceptibility profiles for most other disinfectants. Benzyldimethylammonium chlorides and C10AC were the primary active components in disinfectants DC&R and P-128, respectively, against non-O157 STECs. A disinfectant FS512 MIC ≥ 8 µg/ml was more prevalent among animal O121 strains (61.5%) than among human O121 strains (25%), which may also suggest a source of human exposure to STEC O121 other than food animals. Bacterial inhibition was not dependent solely on pH but was correlated with the presence of dissociated organic acid species and some undissociated acids.

Canonical discrimination of the effect of a new broiler production facility on soil chemical profiles as related to current management practices.

The effect dirt-floored broiler houses have on the underlying native soil, and the potential for contamination of the ground water by leaching under the foundation, is an understudied area. This study examines alterations in fifteen quantitative soil parameters (Ca, Cu, electrical conductivity, Fe, K, Mg, Mn, Na, NO3, organic matter, P, pH, S, soil moisture and Zn) in the underlayment of a newly constructed dirt-floored broiler house over the first two years of production (Native through Flock 11). The experiment was conducted near NW Robertson County, Texas, where the native soil is a fine, smectitic thermic Udertic Paleustalfs and the slopes range from zero to three percent. Multiple samples were collected from under each of three water and three feed lines the length of the house, in a longitudinal study during February 2008 through August 2010. To better define the relationship between the soil parameters and sampling times, a canonical discriminant analysis approach was used. The soil profiles assembled into five distinctive clusters corresponding to time and management practices. Results of this work revealed that the majority of parameters increased over time. The management practices of partial and total house clean-outs markedly altered soil profiles the house underlayment, thus reducing the risk of infiltration into the ground water near the farm. This is important as most broiler farms consist of several houses within a small area, so the cumulative ecological impact could be substantial if not properly managed.

Differential nitric oxide production by chicken immune cells.

Nitric oxide is a rapidly reacting free radical which has cytotoxic effects during inflammatory responses and regulatory effects as a component of signal transduction cascades. We quantified the production of nitrite, a stable metabolite of nitric oxide, in chicken heterophils, monocytes and macrophages after stimulation by IFNgamma, LPS and killed bacteria. Our results demonstrate a differential production of nitrite over 72 h by chicken peripheral blood heterophils, monocytes and the chicken macrophage cell line (HD11). HD11 cells produced an average of 10 fold more nitrite in comparison to monocytes and 30 fold more than heterophils upon stimulation. This production could be inhibited by S-methylisothiourea indicating that the inducible nitric oxide synthase enzyme was participating in the pathway leading to nitrite production.

Campylobacter prevalence in lactating dairy cows in the United States.

The objective of the present study was to determine the prevalence of intestinal Campylobacter in lactating dairy cows from various regions of the United States. Participating commercial dairy farms were chosen at random and were part of a national survey to determine E. coli O157:H7 and Salmonella prevalence in dairy cows. Farms had no previous history of Campylobacter problems. Fecal samples were collected rectally from 720 cows on farms in the northeast (four farms), in the desert southwest (three farms), and in the Pacific west (two farms). A minimum of 60 fecal samples per visit were collected from each farm. Thirty isolates were analyzed using the RiboPrinter Microbial Characterization System to obtain ribosomal RNA patterns. Twenty isolates were tentatively identified as Campylobacter jejuni, two as Campylobacter coli, three as Campylobacter spp., and five as unknown. Individual single-visit farm prevalence ranged from 0 to 10%. The disk diffusion method, employing 11 antibiotics, was used to test the antibiotic sensitivities of 27 of the isolates. Eight isolates were resistant to two or more antibiotics, 13 isolates were resistant to one antibiotic, and 6 were totally susceptible. Under the conditions of this study, the authors conclude that Campylobacter prevalence in lactating dairy cows in the United States is low, there is no difference in prevalence on the basis of geographical location, the predominant species is C. jejuni, and that the majority of these isolates are sensitive to antibiotics.

Cecal colonization of chicks by bovine-derived strains of Campylobacter.

Campylobacter jejuni and Campylobacter coli strains were isolated from feces of dairy cattle at farms with no known problem due to campylobacteria. Farms were located in the northeast, desert southwest, and Pacific west. Twenty isolates were identified by ribotyping with a RiboPrinter. The ability of these bovine isolates to colonize the ceca of chicks was determined by challenge inoculation and reisolation of the challenge strain from the ceca at 1 and 2 wk after challenge. Isolates recovered from chick ceca were examined by ribotyping to assure they matched the challenge strain. One hundred percent of the bovine-derived challenge strains were capable of colonizing chicks. These results indicate that dairy cattle may be asymptomatic Campylobacter carriers and potential sources of campylobacteria contamination of poultry facilities.

Disinfectant and antibiotic susceptibility profiles of Escherichia coli O157:H7 strains from cattle carcasses, feces, and hides and ground beef from the United States.

The disinfectant and antibiotic susceptibility profiles of 344 Escherichia coli O157:H7 strains from cattle carcasses, feces, and hides and ground beef from the United States were determined. A low prevalence of antibiotic resistance was observed (14%). The highest prevalences of resistance were to sulfisoxazole (10.5%), tetracycline (9.9%), streptomycin (7%), and chloramphenicol (4.9%). Four strains were resistant to eight antibiotics (two strains from ground beef and one strain each from hide and preevisceration carcass swabs of cull cattle at harvest). Pulsed-field gel electrophoresis analysis of the E. coli O157:H7 strains revealed two major groups (designated 1 and 2) composed of 17 and 20 clusters, respectively. Clusters 1A, 1B, 1C, and 1G.1 were associated with multidrug-resistant strains. There was no observed correlation between disinfectant resistance and antibiotic resistance. Sixty-nine (20%) of the 344 strains were resistant to chlorhexidine or benzalkonium chloride or the MICs of benzyldimethyldodecylammonium chloride were elevated. Inducible resistance was observed at elevated concentrations of antibiotics (1.4%) and disinfectants (6.1%). The highest rate of disinfectant inducible resistance was to OdoBan, quaternary ammonium chlorides, and the surface disinfectants F25, FS512, and MG, which are used in dairies, restaurants, and food processing plants. High MICs (1,024 to 4,096 m g/ml) of acetic, lactic, and citric acids were found. The decreasing order of acid potency based on molar MICs (MICs(molar)) was acetic, citric, and lactic acid. The correlation of the concentration of dissociated organic acids and MICs(molar) strongly suggests that the observed inhibition of E. coli O157:H7 was primarily due to dissociated forms of the acids.

Destruction of single-species biofilms of Escherichia coli or Klebsiella pneumoniae subsp. pneumoniae by dextranase, lactoferrin, and lysozyme.

The aim of this work was to determine the destructive activity of dextranase, lactoferrin, and lysozyme, against single species biofilms composed of either Klebsiella pneumoniae subsp. pneumoniae or Escherichia coli using the MBEC Assay. Luminescence measurements based on quantitation of the ATP present were used to determine the amount of biofilm elimination and correlated with quantity of live bacteria present in the sample. The data were analyzed employing a two-way ANOVA and Bonferroni post-test. Treatments resulted in percentage reductions of E. coli biofilms ranging from 73 to 98%. Lactoferrin (40 microg/ml) produced a significantly higher-percentage reduction than lysozyme (10 microg/ml) (P < 0.05), no other significant differences occurred. Similar treatments resulted in percentage reductions of K. pneumoniae subsp. pneumoniae biofilms ranging from 51 to 100%. Dextranase treatments produced a significantly lower percentage reduction than all other materials (P < 0.05), no other significant differences occurred. No material was capable of complete destruction of both single species biofilms; however, low concentrations of lactoferrin and lysozyme each removed 100% of the K. pneumoniae subsp. pneumoniae biofilm. Low concentrations of lactoferrin or lysozyme might be beneficial to prevent biofilm formation by K. pneumoniae subsp. pneumoniae.

Evaluation of Salmonella movement through the gut of the lesser mealworm, Alphitobius diaperinus (Coleoptera: Tenebrionidae).

AIMS: The lesser mealworm, Alphitobius diaperinus is an important poultry pest prevalent during production that is capable of vectoring pathogens. This study was undertaken to determine the gut transit time of Salmonella for biosecurity risk analysis of pathogen dispersal into the environment. METHODS: Adult and larval A. diaperinus were exposed to two concentrations of a fluorescently labeled Salmonella enterica for 15, 30, and 60 min time periods then externally disinfected to evaluate internal transfer of Salmonella. The insects were monitored every 30 min over 4 h and evacuated frass (feces) processed for the marker Salmonella. The minimum time monitored was 45 min (15 exposure+30 min time point), and the maximum was 5 h (60 exposure+4 h time point). RESULTS: Adults treated with 10(6) or 10(8) colony-forming units (cfu)/mL, which produced Salmonella positive frass within the 5 h experimental time, displayed a mean gut transit time of 144.4 min (range 90-270 min) and 186.3 min (range 120-300 min), respectively. Larvae treated with 10(6) or 10(8) cfu/mL displayed a mean gut transit time of 172.5 min (range 120-300 min) and 131.7 min (range 60-300 min), respectively. SIGNIFICANCE AND IMPACT OF STUDY: Understanding the sources and contribution of reservoir populations of pathogens in poultry production operations is important for development of biosecurity measures to mitigate their transfer. A. diaperinus are prevalent in production operations and difficult to suppress. Management standards accept the reutilization of litter in which insects survive between flock rotations. Removing litter and spreading it onto nearby fields results in the inadvertent dispersal of beetles. Few studies demonstrating the specific bacterial dispersal capacities of these insects have been performed. This study determined that Salmonella acquired internally, commonly transits the gut, allowed the insect to disperse viable pathogenic bacteria within 2-3 h.