Chronic obstructive pulmonary disease and asthma-associated Proteobacteria, but not commensal Prevotella spp., promote Toll-like receptor 2-independent lung inflammation and pathology.

Recent studies of healthy human airways have revealed colonization by a distinct commensal bacterial microbiota containing Gram-negative Prevotella spp. However, the immunological properties of these bacteria in the respiratory system remain unknown. Here we compare the innate respiratory immune response to three Gram-negative commensal Prevotella strains (Prevotella melaninogenica, Prevotella nanceiensis and Prevotella salivae) and three Gram-negative pathogenic Proteobacteria known to colonize lungs of patients with chronic obstructive pulmonary disease (COPD) and asthma (Haemophilus influenzae B, non-typeable Haemophilus influenzae and Moraxella catarrhalis). The commensal Prevotella spp. and pathogenic Proteobacteria were found to exhibit intrinsic differences in innate inflammatory capacities on murine lung cells in vitro. In vivo in mice, non-typeable H. influenzae induced severe Toll-like receptor 2 (TLR2)-independent COPD-like inflammation characterized by predominant airway neutrophilia, expression of a neutrophilic cytokine/chemokine profile in lung tissue, and lung immunopathology. In comparison, P. nanceiensis induced a diminished neutrophilic airway inflammation and no detectable lung pathology. Interestingly, the inflammatory airway response to the Gram-negative bacteria P. nanceiensis was completely TLR2-dependent. These findings demonstrate weak inflammatory properties of Gram-negative airway commensal Prevotella spp. that may make colonization by these bacteria tolerable by the respiratory immune system.

Evaluation of Steam-Ultrasound Decontamination on Naturally Contaminated Broilers through the Analysis of Campylobacter, Total Viable Count, and Enterobacteriaceae.

ABSTRACT: This study aimed to evaluate the decontamination effects of steam-ultrasound application, through specially designed nozzles installed inside a constructed machine, with a capacity of 10,500 birds per h on naturally contaminated broilers. Using three different skin-sampling areas-back, breast, and neck skin-microbial analysis of Campylobacter, Enterobacteriaceae, and total viable count (TVC) was performed before and after steam-ultrasound treatment. In total, 648 skin samples were analyzed for Campylobacter, and 216 samples were analyzed for Enterobacteriaceae and TVC. Results showed Campylobacter reductions (P < 0.001) of 0.8, 1.1, and 0.7 log, analyzed from back, breast, and the neck skin samples, respectively. Furthermore, reductions of Enterobacteriaceae (P < 0.001) by 1.6, 1.9, and 1.1 log and reductions of TVC (P < 0.001) by 2.0, 2.4, and 1.3 log were found on back, breasts, and neck, respectively. Campylobacter levels were evaluated after 8 days of refrigeration at 4°C in control and steam-ultrasound-treated broilers to determine contamination stability in a small 12-sample trial. The results showed no changes in reductions during refrigeration, indicating that reduced Campylobacter numbers remained stable in treated broilers. This study showed significant bacterial reduction was achieved in three different broiler surface areas at a slaughter speed of 10,500 birds per h at temperatures more than 80°C. The rapid treatment of less than 1.5-s exposure time inside the chamber makes this technology potentially suitable for modern and fast poultry processing lines.

Combined steam-ultrasound treatment of 2 seconds achieves significant high aerobic count and Enterobacteriaceae reduction on naturally contaminated food boxes, crates, conveyor belts, and meat knives.

Food contact surfaces require rigorous sanitation procedures for decontamination, although these methods very often fail to efficiently clean and disinfect surfaces that are visibly contaminated with food residues and possible biofilms. In this study, the results of a short treatment (1 to 2 s) of combined steam (95°C) and ultrasound (SonoSteam) of industrial fish and meat transportation boxes and live-chicken transportation crates naturally contaminated with food and fecal residues were investigated. Aerobic counts of 5.0 to 6.0 log CFU/24 cm(2) and an Enterobacteriaceae spp. level of 2.0 CFU/24 cm(2) were found on the surfaces prior to the treatment. After 1 s of treatment, the aerobic counts were significantly (P < 0.0001) reduced, and within 2 s, reductions below the detection limit (<10 CFU) were reached. Enterobacteriaceae spp. were reduced to a level below the detection limit with only 1 s of treatment. Two seconds of steam-ultrasound treatment was also applied on two different types of plastic modular conveyor belts with hinge pins and one type of flat flexible rubber belt, all visibly contaminated with food residues. The aerobic counts of 3.0 to 5.0 CFU/50 cm(2) were significantly (P < 0.05) reduced, while Enterobacteriaceae spp. were reduced to a level below the detection limit. Industrial meat knives were contaminated with aerobic counts of 6.0 log CFU/5 cm(2) on the handle and 5.2 log CFU/14 cm(2) on the steel. The level of Enterobacteriaceae spp. contamination was approximately 2.5 log CFU on the handle and steel. Two seconds of steam-ultrasound treatment reduced the aerobic counts and Enterobacteriaceae spp. to levels below the detection limit on both handle and steel. This study shows that the steam-ultrasound treatment may be an effective replacement for disinfection processes and that it can be used for continuous disinfection at fast process lines. However, the treatment may not be able to replace efficient cleaning processes used to remove high loads of debris.

Combined steam and ultrasound treatment of broilers at slaughter: a promising intervention to significantly reduce numbers of naturally occurring campylobacters on carcasses.

Steam or hot water decontamination treatment of broiler carcasses is hampered by process limitations due to prolonged treatment times and adverse changes to the epidermis. In this study, a combination of steam with ultrasound (SonoSteam®) was investigated on naturally contaminated broilers that were processed at conventional slaughter speeds of 8,500 birds per hour in a Danish broiler plant. Industrial-scale SonoSteam equipment was installed in the evisceration room, before the inside/outside carcass washer. The SonoSteam treatment was evaluated in two separate trials performed on two different dates. Numbers of naturally occurring Campylobacter spp. and TVC were determined from paired samples of skin excised from opposite sides of the breast of the same carcass, before and after treatments. Sampling was performed at two different points on the line: i) before and after the SonoSteam treatment and ii) before the SonoSteam treatment and after 80 min of air chilling. A total of 44 carcasses were examined in the two trials. Results from the first trial showed that the mean initial Campylobacter contamination level of 2.35 log10 CFU was significantly reduced (n=12, p<0.001) to 1.40 log10 CFU after treatment. A significant reduction (n=11, p<0.001) was also observed with samples analyzed before SonoSteam treatment (2.64 log10 CFU) and after air chilling (1.44 log10 CFU). In the second trial, significant reductions (n=10, p<0.05) were obtained for carcasses analyzed before (mean level of 2.23 log10 CFU) and after the treatment (mean level of 1.36 log10 CFU). Significant reductions (n=11, p<0.01) were also found for Campylobacter numbers analyzed before the SonoSteam treatment (2.02 log10 CFU) and after the air chilling treatment (1.37 log10 CFU). The effect of air chilling without SonoSteam treatment was determined using 12 carcasses pre- and postchill. Results showed insignificant reductions of 0.09 log10 from a mean initial level of 2.19 log10 CFU. Numbers of TVC before treatments ranged between 3.47 and 4.79 log10 CFU. In all cases, TVC was significantly (p<0.001, n=45 in each trial) reduced by approximately 0.7 log10 CFU. An authorized sensory panel at the Danish Veterinary and Food Administration concluded that broiler carcasses treated with SonoSteam were acceptable for purchase. These conclusions were based on organoleptic differences (smell, skin/meat consistency, texture and color) of treated vs. untreated carcasses. Results obtained from this study suggest that steam-ultrasound treatment of carcasses at broiler processing plants can significantly reduce numbers of Campylobacter on naturally contaminated broilers.