Modelling-based identification of factors influencing campylobacters in chicken broiler houses and on carcasses sampled after processing and chilling.

AIMS: To identify production and processing practices that might reduce Campylobacter numbers contaminating chicken broiler carcasses. METHODS AND RESULTS: The numbers of campylobacters were determined on carcass neck skins after processing or in broiler house litter samples. Supplementary information that described farm layouts, farming conditions for individual flocks, the slaughterhouse layouts and operating conditions inside plants was collected, matched with each Campylobacter test result. Statistical models predicting the numbers of campylobacters on neck skins and in litter were constructed. Carcass microbial contamination was more strongly influenced by on-farm production practices compared with slaughterhouse activities. We observed correlations between the chilling, washing and defeathering stages of processing and the numbers of campylobacters on carcasses. There were factors on farm that also correlated with numbers of campylobacters in litter. These included bird gender, the exclusion of dogs from houses, beetle presence in the house litter and the materials used to construct the house frame. CONCLUSIONS: Changes in farming practices have greater potential for reducing chicken carcass microbial contamination compared with processing interventions. SIGNIFICANCE AND IMPACT OF THE STUDY: Routine commercial practices were identified that were correlated with lowered numbers of campylobacters. Consequently, these practices are likely to be both cost-effective and suitable for adoption into established farms and commercial processing.

Two-photon absorption measurements of deep UV transmissible materials at 213 nm.

We report on two-photon absorption measurements at 213 nm of deep UV transmissible media, including LiF, MgF2, CaF2, BaF2, sapphire (Al2O3), and high-purity grades of fused-silica (SiO2). A high-stability 24 ps Nd:YAG laser operating at the 5th harmonic (213 nm) was used to generate a high-intensity, long-Rayleigh-length Gaussian focus inside the samples. The measurements of the fluoride crystals and sapphire indicate two-photon absorption coefficients between 0.004 and 0.82 cm/GW. We find that different grades of fused silica performed near identically for two-photon absorption; however, there are differences in linear losses associated with purity. A low two-photon absorption cross section is measured for MgF2, making it an ideal material for the propagation of high-intensity deep UV lasers.

Site-dependent changes in structure and function of lapine articular cartilage 4 weeks after anterior cruciate ligament transection.

OBJECTIVE: The aim of this study was to investigate the site-dependent changes in the structure and function of articular cartilage in the lapine knee joint at a very early stage of osteoarthritis (OA), created experimentally by anterior cruciate ligament transection (ACLT). METHODS: Unilateral ACLT was performed in eight mature New Zealand white rabbits. ACL transected and contralateral (C-L) joints were prepared for analysis at 4 weeks after ACLT. Three rabbits with intact joints were used as a control group (CNTRL). Femoral groove, medial and lateral femoral condyles, and tibial plateaus were harvested and used in the analysis. Biomechanical tests, microscopy and spectroscopy were used to determine the biomechanical properties, composition and structure of the samples. A linear mixed model was chosen for statistical comparisons between the groups. RESULTS: As a result of ACLT, the equilibrium and dynamic moduli were decreased primarily in the femoral condyle cartilage. Up to three times lower moduli (P < 0.05) were observed in the ACLT group compared to the control group. Significant (P < 0.05) proteoglycan (PG) loss in the ACLT joint cartilage was observed up to a depth of 20-30% from the cartilage surface in femoral condyles, while significant PG loss was confined to more superficial regions in tibial plateaus and femoral groove. The collagen orientation angle was increased (P < 0.05) up to a cartilage depth of 60% by ACLT in the lateral femoral condyle, while smaller effects, but still significant, were observed at other locations. The collagen content was increased (P < 0.05) in the middle and deep zones of the ACLT group compared to the control group samples, especially in the lateral femoral condyle. CONCLUSION: Femoral condyle cartilage experienced the greatest structural and mechanical alterations in very early OA, as produced by ACLT. Degenerative alterations were observed especially in the superficial collagen fiber organization and PG content, while the collagen content was increased in the deep tissue of femoral condyle cartilage. The current findings provide novel information of the early stages of OA in different locations of the knee joint.

To fast or feed: an alternative life history for anadromous brook trout Salvelinus fontinalis overwintering within a harbour.

The seasonal feeding pattern of sea-run brook trout Salvelinus fontinalis was studied from November to May 2010-2012 in Antigonish Harbour, Nova Scotia, Canada (45° 38' N; 61° 55' W). Sixty-three S. fontinalis (mean ± s.d. fork length = 330 ± 70 mm and mass = 536 ± 351 g) captured had fed predominantly on fishes (Fundulidae and Gasterosteidae). Percentage of empty stomachs was highest during autumn (18%) and winter (22%) and lowest in spring (7%). Stomach fullness increased from autumn to a maximum during winter, relating to near-zero body temperatures which may have effectively stopped gastric evacuation. Although feeding occurred during winter (December to March), consumption rates were calculated as negative values, and subsequently returned to positive values in spring (April to May). The over-winter life-history strategy of this sea-run S. fontinalis population appears to be a feeding marine migration in which fish continually increase body condition, representing an alternative to the more common overwintering strategy of starvation in fresh water until spring.

Spinal cord vascular degeneration impairs duloxetine penetration.

Introduction: Chronic pain is a prevalent physically debilitating health-related morbidity. Frontline analgesics are inadequate, providing only partial pain relief in only a proportion of the patient cohort. Here, we explore whether alterations in spinal cord vascular perfusion are a factor in reducing the analgesic capability of the noradrenaline reuptake inhibitor, duloxetine. Method: An established rodent model of spinal cord vascular degeneration was used. Endothelial-specific vascular endothelial growth factor receptor 2 knockout mouse was induced via hydroxytamoxifen administered via intrathecal injection. Duloxetine was administered via intraperitoneal injection, and nociceptive behavioural testing was performed in both WT and VEGFR2KO mice. LC-MS/MS was performed to explore the accumulation of duloxetine in the spinal cord in WT and VEGFR2KO mice. Results: Spinal cord vascular degeneration leads to heat hypersensitivity and a decline in capillary perfusion. The integrity of noradrenergic projections (dopa - hydroxylase labelled) in the dorsal horn remained unaltered in WT and VEGFR2KO mice. There was an association between dorsal horn blood flow with the abundance of accumulated duloxetine in the spinal cord and analgesic capacity. In VEGFR2KO mice, the abundance of duloxetine in the lumbar spinal cord was reduced and was correlated with reduced anti-nociceptive capability of duloxetine. Discussion: Here, we show that an impaired vascular network in the spinal cord impairs the anti-nociceptive action of duloxetine. This highlights that the spinal cord vascular network is crucial to maintaining the efficacy of analgesics to provide pain relief.

Quantitative Determination of Campylobacter on Broilers along 22 United Kingdom Processing Lines To Identify Potential Process Control Points and Cross-Contamination from Colonized to Uncolonized Flocks.

ABSTRACT: As part of a program to reduce numbers of the human pathogen Campylobacter on retail chickens, 22 broiler processing lines, representing more than 90% of UK production, were characterized by enumerating Campylobacter on pooled neck skins after exsanguination, scalding, defeathering, evisceration, crop removal, inside-outside washing, and air-chilling stages of processing. Sixteen of the processing lines investigated showed significant (P < 0.05) reductions in Campylobacter numbers because of carcass scalding. However, in all of these lines, the following defeathering stage caused a significant increase in Campylobacter contamination that effectively negated the reductions caused by scalding. On four processing lines, primary chilling also caused a significant reduction in numbers of Campylobacter. On three lines, there was a significant microbiological benefit from inside-outside washing. The stages where Campylobacter numbers were reduced require further investigation to determine the specific mechanisms responsible so that the observed pathogen reductions can be optimized and then more widely implemented. The transfer of up to 4 log CFU Campylobacter per g of neck skin from a colonized flock to a following uncolonized flock was observed. Cross-contamination was substantial and still detectable after 5,000 carcasses from an uncolonized flock had been processed. Numbers of Campylobacter recovered from the uncolonized flocks were highest on the first of the uncolonized birds to pass along the line, and in general, the numbers declined as more uncolonized birds were processed. Air sampling recovered low numbers at the processing stages monitored, indicating that airborne transmission was unlikely to be the primary transfer mechanism operating for cross-contamination between flocks.

Epidemiology and outcomes of primary sclerosing cholangitis: an Australian multicentre retrospective cohort study.

BACKGROUND AND AIMS: Little is known regarding the epidemiology and outcomes of patients with primary sclerosing cholangitis (PSC) in Australia. We, therefore, evaluated the epidemiology and clinical outcomes of PSC in a large cohort of Australian patients and compared these to the general population. METHODS: We conducted a multicentre, retrospective cohort study of PSC patients at nine tertiary liver centers across three Australian states, including two liver transplant centers. RESULTS: A total of 413 PSC patients with 3,285 person-years of follow-up were included. Three hundred and seventy-one (90%) patients had large duct PSC and 294 (71%) had associated inflammatory bowel disease. A total of 168 (41%) patients developed cirrhosis (including 34 at the time of PSC diagnosis) after a median of 15.8 (95% CI 12.4, NA) years. The composite endpoint of death or liver transplantation occurred in 49 (12%) and 78 (19%) patients, respectively, with a median transplant-free survival of 13.4 (95% CI 12.2-15) years. Compared to the general population, PSC accounted for a 240-fold increased risk of development of cholangiocarcinoma (CCA) and CCA-related death. CCA risk was increased with older age of PSC diagnosis, presence of dominant stricture and colectomy. Compared to same-aged counterparts in the general population, PSC patients who were diagnosed at an older age or with longer disease duration had reduced relative survival. CONCLUSION: In this large retrospective cohort study of PSC patients in Australia, increased age and time from diagnosis was associated with increased mortality and morbidity particularly from CCA and development of cirrhosis, necessitating need for liver transplant.

Restoring the selectivity of Bolton broth during enrichment for Campylobacter spp. from raw chicken.

AIMS: When isolating Campylobacter spp. from retail raw chicken using BS EN ISO 10272-1:2006, contaminants frequently cause overgrowth on mCCDA plates. Therefore, these organisms proliferate in the enrichment medium, Bolton broth, indicating a lack of selectivity in this medium. This study sought to characterize the contaminant flora and to devise a modified Bolton broth to inhibit their growth. METHODS AND RESULTS: Contaminants (n=30) from separate samples were identified and antibiotic resistances determined. Most (93%) were extended spectrum ß-lactamase (ESBL) producing Escherichia coli, able to hydrolyse the cefoperazone present in Bolton broth and mCCDA. To inhibit these organisms, original formulation Bolton broth was supplemented with potassium clavulanate, at three concentrations, and recoveries of campylobacters from raw chicken were determined. Using standard Bolton broth, only 49% of samples (n=104) yielded campylobacters, but supplementation with 2 mg l(-1) potassium clavulanate increased this significantly (P<0.05), with 91% of samples positive. CONCLUSIONS: Potassium clavulanate can restore the selectivity of Bolton broth when isolating Campylobacter spp. from raw chicken. SIGNIFICANCE AND IMPACT OF THE STUDY: Raw chicken is often contaminated with the pathogen Campylobacter, but the ISO methodology for its detection is becoming compromised by the increasing presence of antibiotic-resistant bacteria. A simple modification ensures effective detection of this pathogen.

Pyrosequencing and characterization of immune response genes from the American dog tick, Dermacentor variabilis (L.).

Ticks continue to be a threat to animal and human health, and new and novel control strategies are needed for ticks and tick-borne pathogens. The characterization of the tick-pathogen interface and the tick immune response to microbial infections is fundamental toward the formulation of new control strategies for ticks and the pathogens they transmit. Our overall hypothesis for this research is that the tick immune system manages the maintenance of pathogens. Therefore, discovery of tick immune response genes may provide targets for novel control strategies directed toward reducing vector competency and pathogen transmission. In these studies, 454 pyrosequencing, a high-throughput genomic sequencing method was used to discover tick genes expressed in response to bacterial and fungal infections. Expressed sequence tags (ESTs) were analysed from Dermacentor variabilis ticks that had been injected with bacteria (Escherichia coli, Bacillus subtilis, Micrococcus luteus) or fungi (Saccharomyces cerevisiae and Candida albicans) and ticks that were naturally infected with the intracellular bacterium, Anaplasma marginale. By this approach, ESTs were assembled into 5995 contigs. Contigs fell into the five main functional categories of metabolism, genetic information processing, environmental information processing, cellular processes and human diseases. We identified more than 30 genes that are likely to encode for proteins involved in tick immune function. We further analysed by reverse transcriptase PCR (RT-PCR) the expression of 22 of these genes in each of our bacterial or fungal treatment groups and found that seven were up-regulated. Up-regulation of these seven genes was confirmed for bacterial, but not fungal treatment by quantitative PCR (qPCR). One of these products was novel, encoding a new tick defensin. Our results clearly demonstrate the complexities of the tick immune system and mark new directions for further study and characterization of proteins that modulate microbial infections in the American dog tick.

Detecting Climate Change due to Increasing Carbon Dioxide.

The observed interannual variability of temperature at 60 degrees N has been investigated. The results indicate that the surface warming due to increased carbon dioxide which is predicted by three-dimensional climate models should be detectable now. It is not, possibly because the predicted warming is being delayed more than a decade by ocean thermal inertia, or because there is a compensating cooling due to other factors. Further consideration of the uncertainties in model predictions and of the likely delays introduced by ocean thermal inertia extends the range of time for the detection of warming, if it occurs, to the year 2000. The effects of increasing carbon dioxide should be looked for in several variables simultaneously in order to minimize the ambiguities that could result from unrecognized compensating cooling.

Use of conductimetry to rapidly determine relative stress sensitivity in Salmonella isolates.

AIMS: To compare conventional plate counting and indirect conductimetry as techniques for ranking the resistance of Salmonella spp. to processing stressors. METHODS AND RESULTS: Forty Salmonella isolates were subjected to three separate stressors used in food processing; irradiation, heat and high hydrostatic pressure (HHP). Total viable counts (TVC) using conventional plate counts and time to detection (TTD) using indirect conductimetry were determined. A significant negative correlation between TVC and TTD was seen with irradiation (P < 0.01) and heat (P < 0.05) but not HHP. CONCLUSIONS: For a group of salmonellas, indirect conductimetry can rapidly determine a ranking of isolate sensitivity to irradiation and heat. However, for HHP, the results indicated that conventional plate counting alone cannot be used to determine sensitivity. SIGNIFICANCE AND IMPACT OF THE STUDY: The resistance of micro-organisms to processing systems must be ranked to allow the selection of appropriate isolates for process validation. TTD measurements allow rapid screening of salmonellas to rank isolates for resistance to irradiation and heat stress. However, following HHP, the TVC of survivors is independent of the time required for growth to a set cell density and therefore it cannot be used as the sole measure of relative stress resistance.

Factors affecting the recovery of Campylobacter spp. from retail packs of raw, fresh chicken using ISO 10272-1:2006.

AIMS: This study sought to determine the most effective protocol for the detection of Campylobacter spp. in retail packs of fresh, raw chicken based on ISO 10272-1:2006. METHODS AND RESULTS: Three sample preparation protocols were studied; two based on excision and one combining excision with a rinse of the remaining sample. Enrichment cultures were incubated both in closed bottles and microaerobically, and sub-cultured at 24 and 48 h. Packs of chicken (110) were analysed and only two yielded no Campylobacter spp. Subculturing enrichment broths at 24 h gave the same prevalence as at 48 h, P > 0.4 but microaerobic incubation yielded approximately 50% more positive samples than did incubation in closed bottles. Sampling based on excision plus rinsing gave the highest Campylobacter prevalence (92.7%). CONCLUSIONS: To isolate Campylobacter spp. from retail packs of chicken, enrichment cultures must be incubated in a microaerobic atmosphere and sub-cultured at 24 h and, possibly, 48 h. Sampling packs by excision plus rinsing maximized recoveries. SIGNIFICANCE AND IMPACT OF THE STUDY: ISO 10272-1:2006 permits the use of inefficient protocols which markedly underestimate the true prevalence of Campylobacter spp. in retail, fresh chicken. Equivalent results could be obtained 24 h earlier, with consequent savings. Its revision is essential.

Consequences of Using Two Types of Skin Samples from Chilled Chicken Broiler Carcasses To Measure the Degree of Contamination by Campylobacter spp.

HIGHLIGHTS: Campylobacter levels on chicken neck and breast skin were compared. Neck skin was significantly more contaminated (P < 0.05) than breast skin. No relationship between the two skin types was found for Campylobacter levels. A UK government reduction target for highly contaminated chicken was not achieved.

Use of Hugh and Leifson's medium as a simple screening test to aid in the differentiation of Arcobacter spp. from background flora during their isolation from foodstuffs.

AIMS: To investigate the suitability of Hugh and Leifson's medium (HLM) as the basis of a simple screening test to differentiate between contaminants and Arcobacter spp. during their isolation from foodstuffs. METHODS AND RESULTS: Characterized Arcobacter spp. were obtained from recognized culture collections. Wild-type isolates of Arcobacter spp. and contaminants were obtained using published isolation protocols. Retail packs of red meats were used as the source of the isolates. Eighteen defined Arcobacter spp. gave no reaction on HLM, as did 10 local wild-type isolates. Overall 163 contaminants were studied for oxidative reactions on HLM and 86% of isolates demonstrated this property. CONCLUSIONS: HLM can usefully serve as a simple and effective screening test to differentiate between Arcobacter spp. and contaminants. SIGNIFICANCE AND IMPACT OF THE STUDY: Arcobacter isolation procedures are still being developed, and no effective diagnostic media currently exist. Rapidly excluding most contaminants can markedly increase the efficiency of isolation procedures by removing the need for extensive biotyping or the requirement to isolate DNA and conduct PCR tests.

Anomalous material-dependent transport of focused, laser-driven proton beams.

Intense lasers can accelerate protons in sufficient numbers and energy that the resulting beam can heat materials to exotic warm (10 s of eV temperature) states. Here we show with experimental data that a laser-driven proton beam focused onto a target heated it in a localized spot with size strongly dependent upon material and as small as 35 µm radius. Simulations indicate that cold stopping power values cannot model the intense proton beam transport in solid targets well enough to match the large differences observed. In the experiment a 74 J, 670 fs laser drove a focusing proton beam that transported through different thicknesses of solid Mylar, Al, Cu or Au, eventually heating a rear, thin, Au witness layer. The XUV emission seen from the rear of the Au indicated a clear dependence of proton beam transport upon atomic number, Z, of the transport layer: a larger and brighter emission spot was measured after proton transport through the lower Z foils even with equal mass density for supposed equivalent proton stopping range. Beam transport dynamics pertaining to the observed heated spot were investigated numerically with a particle-in-cell (PIC) code. In simulations protons moving through an Al transport layer result in higher Au temperature responsible for higher Au radiant emittance compared to a Cu transport case. The inferred finding that proton stopping varies with temperature in different materials, considerably changing the beam heating profile, can guide applications seeking to controllably heat targets with intense proton beams.

Salmonella in slaughter pigs in Northern Ireland: prevalence and use of statistical modelling to investigate sample and abattoir effects.

A cross-sectional survey of pigs at slaughter in Northern Ireland was undertaken to determine the overall prevalence of Salmonella infection. In total 513 pigs were sampled across four abattoirs, with Salmonella spp. isolated from the caecal contents of 31.4% (95% confidence interval [CI] 27.4%-35.4%) and from 40.0% (95% CI 35.8%-44.3%) of swabs taken from the surface of carcasses post-evisceration. Two serovars, S. Typhimurium and S. Derby, were predominant and accounted for 52% and 35% respectively, of isolates from caecal contents. Antimicrobial resistance was most common amongst isolates of S. Typhimurium with 63.9% multiresistant compared to 10.8% of S. Derby isolates and 8.0% of other Salmonella spp. The proportion of pigs showing serological evidence of infection was significantly lower, with 11.5% (95% CI 8.9%-14.6%) and 10.1% (95% CI 7.7%-13.1%) of meat-juice samples giving positive and suspect reactions, respectively. The ratio of caecal positive to serologically positive animals is higher than in a number of other studies and may suggest recent infection, such as infection occurring during transport or lairage, in a proportion of animals. Statistical (logistic regression) modelling was used to investigate the association between the risk of Salmonella on carcasses and the isolation of Salmonella from caecal contents, and/or the serological status of the animal, while adjusting for other possible explanatory and confounding variables such as abattoir, season, day and time of sampling. The occurrence of Salmonella in caecal contents (odds ratio [OR] 2.39; 95% CI 1.52-3.77) or a suspect/positive serological reaction (OR 2.15; 95% CI 1.28-3.61) were both independently associated with the occurrence of Salmonella on carcasses in homebred, but interestingly not in imported animals. In most multivariable models there were also significant differences in carcass contamination between seasons with the highest odds of carcass contamination occurring in the April to June quarter and the lowest in the October to December quarter. Differences between sampling days were also evident with the highest odds of carcass contamination at the end of the week (Fridays) and the lowest at the start of the week (Mondays). These associations, after adjusting for the caecal or serological result, would suggest the occurrence of abattoir effects, such varying residual levels of abattoir contamination, which are independent of the individual pig status.

Predictors of maternal mortality and near-miss maternal morbidity.

OBJECTIVE: To identify risk factors for life-threatening maternal outcomes. STUDY DESIGN: Hospital charts were reviewed for cases of maternal mortality or near-miss and for controls overmatched 1:3. Significant risk factors were identified through simple and best subsets multiple logistic regression. RESULT: Eight cases of mortality and 69 near-miss cases were found. Significant risk factors with their odds ratios and 95% confidence intervals are: age 35 to 39 years (2.3, 1.2 to 4.4) and >39 years (5.1, 1.8 to 14.4); African-American race (7.4, 2.5 to 22.0) and Hispanic ethnicity (4.2, 1.3 to 13.2); chronic medical condition (2.7, 1.5 to 4.8); obesity (3.0, 1.7 to 5.3); prior cesarean (5.2, 2.8 to 9.8) and gravidity (1.2, 1.1 to 1.5 per pregnancy). In multivariable logistic regression, race remained significant while controlling for other significant factors and markers of socioeconomic status. CONCLUSION: Some risk factors can be modified through medical care, education or social support systems. Racial disparity in outcome is confirmed and is unexplained by traditional risk factors.

Total body irradiation, fludarabine, melphalan, and allogeneic hematopoietic stem cell transplantation for advanced pediatric hematologic malignancies.

We evaluated the efficacy and toxicity of adding 9 Gy of total body irradiation (TBI), in three single daily fractions of 3 Gy, to the reduced intensity regimen of fludarabine 30 mg/m2 i.v. x 4 days and melphalan 140 mg/m2 i.v. x 1 day in advanced pediatric hematologic malignancies. Twenty-two acute lymphoblastic leukemia (ALL), six acute myeloid leukemia (AML), and one non-Hodgkin lymphoma patients were transplanted. Of these, 13 were beyond second remission, and five had prior hematopoietic stem cell transplant (HSCT). Twenty-one donors were unrelated, of which 19 were from cord blood (CB) units. Three of the eight related donors were genotypically disparate. Oral mucositis and diarrhea were the most common toxicities. Twenty-seven patients achieved neutrophil engraftment (median 16 days), and 23 had platelet engraftment (median 42 days). One patient had primary graft failure. Seven patients died of non-relapse causes in the first 100 days. With a median follow-up of 52 months, seven of 22 ALL, five of six AML, and one of one lymphoma patients are alive and in remission. The regimen of TBI, fludarabine, and melphalan allows the engraftment of allogeneic hematopoietic stem cells (including mismatched CB). It was fairly well tolerated in pediatric patients, even for second transplants. Its efficacy requires further evaluation.

Evaluation of the use of conductimetry for the rapid and precise measurement of Salmonella spp. growth rates.

The growth rates of 14 Salmonella serovars in tryptone soy broth plus yeast extract (TSBYE) were estimated using conventional plating techniques and indirect conductimetry using a Don Whitley RABIT system. Both methods gave identical results for the maximum specific growth rate (mumax) P>0.05. However, using the conductimetric method, mumax for a single serovar was determined in less than 7 h, whereas the conventional method required an additional 24 h. In addition, the conductimetric method was considerably more precise, much less labour-intensive and required the use of considerably less consumables. Using conductimetry, a trained operator could accurately determine mumax for 24 serovars in 3 working days, but only one serovar using the conventional plate counting technique. Hence, the use of conductimetry can markedly increase the precision and accuracy of mumax determinations by allowing a very significant increase in the number of results obtained and in their precision. The data generated will allow the development of better mathematical growth models. The method can also be used to compare growth media and conditions and hence rapidly optimise detection protocols for this pathogen.