Immunomodulatory drugs in sepsis: a systematic review and meta-analysis.

Dysregulation of the host immune response has a central role in the pathophysiology of sepsis. There has been much interest in immunomodulatory drugs as potential therapeutic adjuncts in sepsis. We conducted a systematic review and meta-analysis of randomised controlled trials evaluating the safety and clinical effectiveness of immunomodulatory drugs as adjuncts to standard care in the treatment of adults with sepsis. Our primary outcomes were serious adverse events and all-cause mortality. Fifty-six unique, eligible randomised controlled trials were identified, assessing a range of interventions including cytokine inhibitors; anti-inflammatories; immune cell stimulators; platelet pathway inhibitors; and complement inhibitors. At 1-month follow-up, the use of cytokine inhibitors was associated with a decreased risk of serious adverse events, based on 11 studies involving 7138 patients (RR (95%CI) 0.95 (0.90-1.00), I2 = 0%). The only immunomodulatory drugs associated with an increased risk of serious adverse events were toll-like receptor 4 antagonists (RR (95%CI) 1.18 (1.04-1.34), I2 = 0% (two trials, 567 patients)). Based on 18 randomised controlled trials, involving 11,075 patients, cytokine inhibitors reduced 1-month mortality (RR (95%CI) 0.88 (0.78-0.98), I2 = 57%). Mortality reduction was also shown in the subgroup of 13 randomised controlled trials that evaluated anti-tumour necrosis factor α interventions (RR (95%CI) 0.93 (0.87-0.99), I2 = 0%). Anti-inflammatory drugs had the largest apparent effect on mortality at 2 months at any dose (two trials, 228 patients, RR (95%CI) 0.64 (0.51-0.80), I2 = 0%) and at 3 months at any dose (three trials involving 277 patients, RR (95%CI) 0.67 (0.55-0.81), I2 = 0%). These data indicate that, except for toll-like receptor 4 antagonists, there is no evidence of safety concerns for the use of immunomodulatory drugs in sepsis, and they may show some short-term mortality benefit for selected drugs.

Practitioners' perspectives on health in Strategic Environmental Assessment of spatial planning policies in Scotland.

OBJECTIVES: Local authorities in Scotland are required to produce a Local Development Plan (LDP), which allocates sites for development and sets policies to guide decisions on planning applications. As part of this, local authorities must undertake a Strategic Environmental Assessment (SEA). This is a structured assessment of likely environmental impacts, which includes human health. This study explores how SEA practitioners and SEA consultation authorities consider health. STUDY DESIGN: Qualitative study design using eight in-depth semi-structured interviews. METHODS: Individual interviews were carried out with SEA practitioners from six local authority areas in Scotland and two SEA consultation authorities. Interviews were recorded, transcribed and analysed thematically. RESULTS: Respondents articulated a broad perspective on health, but this was not reflected in SEA practice. Barriers to considering health more fully in SEA included low confidence in assessing health, limited partnership working with public health professionals and the lack of a consultation authority able to cover all aspects of health. Respondents valued partnership work between public health and planning professionals. CONCLUSION: This study suggests recent work in Scotland to increase understanding of the role of spatial planning to influence health has been successful. However, further work is required to expand this to include links between spatial planning and health inequalities. SEA in Scotland does not currently support holistic consideration of health and health inequalities. Strong partnership working between public health and other sectors can increase understanding of links with health and create healthy places.

Conventional oxygen therapy versus CPAP as a ceiling of care in ward-based patients with COVID-19: a multi-centre cohort evaluation.

BACKGROUND: Continuous positive airway pressure (CPAP) therapy is commonly used for respiratory failure due to severe COVID-19 pneumonitis, including in patients deemed not likely to benefit from invasive mechanical ventilation (nIMV). Little evidence exists demonstrating superiority over conventional oxygen therapy, whilst ward-level delivery of CPAP presents practical challenges. We sought to compare clinical outcomes of oxygen therapy versus CPAP therapy in patients with COVID-19 who were nIMV. METHODS: This retrospective multi-centre cohort evaluation included patients diagnosed with COVID-19 who were nIMV, had a treatment escalation plan of ward-level care and clinical frailty scale ≤ 6. Recruitment occurred during the first two waves of the UK COVID-19 pandemic in 2020; from 1st March to May 31st, and from 1st September to 31st December. Patients given CPAP were compared to patients receiving oxygen therapy that required FiO2 ≥0.4 for more than 12 hours at hospitals not providing ward-level CPAP. Logistic regression modelling was performed to compare 30-day mortality between treatment groups, accounting for important confounders and within-hospital clustering. FINDINGS: Seven hospitals provided data for 479 patients during the UK COVID-19 pandemic in 2020. Overall 30-day mortality was 75.6% in the oxygen group (186/246 patients) and 77.7% in the CPAP group (181/233 patients). A lack of evidence for a treatment effect persisted in the adjusted model (adjusted odds ratio 0.84 95% CI 0.57-1.23, p=0.37). 49.8% of patients receiving CPAP-therapy (118/237) chose to discontinue it. INTERPRETATION: No survival difference was found between using oxygen alone or CPAP to treat patients with severe COVID-19 who were nIMV. A high patient-initiated discontinuation rate for CPAP suggests a significant treatment burden. Further reflection is warranted on the current treatment guidance and widespread application of CPAP in this setting. FUNDING: L Pearmain is supported by the MRC (MR/R00191X/1). TW Felton is supported by the NIHR Manchester Biomedical Research Centre.

Effect of Feeding a Postbiotic Derived from Saccharomyces cerevisiae Fermentation as a Preharvest Food Safety Hurdle for Reducing Salmonella Enteritidis in the Ceca of Layer Pullets.

ABSTRACT: Salmonella Enteritidis is responsible for a significant proportion of foodborne salmonellosis in the United States and continues to be attributable to table eggs despite increased federal oversight. Technologies, including feed additives, continue to be evaluated for preharvest application and their potential food safety benefits. Diamond V Original XPC, a Saccharomyces cerevisiae fermentation-based postbiotic (SCFP), was evaluated for its effectiveness in reducing Salmonella Enteritidis (SE) colonization in young layer pullets. A total of 40 day-old Hy-Line W-36 layer pullets were equally divided and randomly assigned to one of two dietary treatments, with SCFP or without SCFP (PCON), and orally gavaged on day 28 with SE at 106 CFU/mL. Another 20 day-old pullets were fed the same control feed without SCFP and blank inoculated on day 28 with 1 mL of sterile phosphate-buffered saline to serve as a negative control. Qualitative and quantitative analyses of cecal contents for Salmonella were performed for all birds on day 32. The prevalence of SE in the ceca of all directly challenged birds was 100%; however, the SE concentration in birds fed SCFP diet (3.35 log CFU/g) was significantly lower (P < 0.0001) than that of the PCON birds not fed SCFP (4.49 log CFU/g). The proportion of birds with enumerable SE concentrations was lower in SCFP-fed pullets (57.9%) than in the PCON pullets (95.0%). These data suggest that inclusion of SCFP in the diet may aid in the reduction of SE within the ceca of commercial laying hens and could serve as an additional preharvest food safety hurdle.

Antibiotic-resistant Staphylococcus aureus strains of swine origin: molecular typing and susceptibility to oregano (Origanum vulgare L.) essential oil and maqui (Aristotelia chilensis (Molina) Stuntz) extract.

AIM: The molecular typing and the susceptibility of Staphylococcus aureus strains of swine origin to antibiotics, oregano (Origanum vulgare L.) essential oil (EO) and Chilean blackberry maqui (Aristotelia chilensis (Molina) Stuntz) extract were determined. METHODS AND RESULTS: Twenty S. aureus strains of swine origin were subjected to molecular typing, of which six strains were selected for antimicrobial susceptibility testing. The epsilon test (Etest) was used to determine the antibiotic susceptibility. The susceptibility to natural antimicrobials (NAs): oregano EO, maqui extract, thymol (Thy) and carvacrol (Carv), was carried out using the disk diffusion method. The S. aureus strains were genetically diverse. All strains were resistant to at least one class of antibiotic, and two strains were multidrug-resistant. The minimum inhibitory concentration of oregano EO, Thy and Carv was 0·01-0·04%. Maqui extract did not show antistaphylococcal activity. CONCLUSIONS: Natural antimicrobials extracted from oregano have an inhibitory activity against S. aureus strains from swine origin, with no effect using maqui extract. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information about the characteristics of S. aureus strains of swine origin, and about the potential use of NAs from oregano to enhance the control of antibiotic-resistant S. aureus strains in the pork supply chain.

Changes in the prevalence of Salmonella serovars associated swine production and correlations of avian, bovine and swine-associated serovars with human-associated serovars in the United States (1997-2015).

As Salmonella enterica is an important pathogen of food animals, surveillance programmes for S. enterica serovars have existed for many years in the United States. Surveillance programmes serve many purposes, one of which is to evaluate alterations in the prevalence of serovars that may signal changes in the ecology of the target organism. The primary aim of this study was to evaluate changes in the proportion of S. enterica serovars isolated from swine over a near 20-year observation period (1997-2015) using four longitudinal data sets from different food animal species. The secondary aim was to evaluate correlations between changes in S. enterica serovars frequently recovered from food animals and changes in S. enterica serovars associated with disease in humans. We found decreasing proportions of S. enterica serovar Typhimurium, serovar Derby and serovar Heidelberg and increasing proportions of S. enterica serovar 4,[5],12:i:-, serovar Infantis and serovar Johannesburg in swine over time. We also found positive correlations for the yearly changes in S. enterica serovar 4,[5],12:i:-, serovar Anatum and serovar Johannesburg between swine and human data; in S. enterica Worthington between avian and human data; and in S. enterica serovar 4,[5],12:i:- between bovine and human data. We found negative correlations for the yearly changes in S. enterica serovar 4,[5],12:i:- and serovar Johannesburg between avian and human data.

An approach to treating a patient with a HeartMate II™ left ventricularassist device in a multiplace hyperbaric chamber: a case report.

INTRODUCTION: Left-ventricular assist devices (LVAD) are becoming a common therapy for end-stage heart failure. These devicesare not tested for pressurization in a hyperbaricchamber by the manufacturer. In this article, we present an approach to modify the power supply in order to safely treat a patient with an LVAD. MATERIALS AND METHODS: Our patient had a HeartMateII™ LVAD and presented for hyperbaric oxygen treatments for severe radiation cystitis. In order to modify this patient's equipment to be compliant with NFPA6 safety standards we made several modifications. In brief, this included eliminating the usage of lithium-ion batteries, modifying the cord to be compatible with Fink chamber outlets, and enclosing the power module in a nitrogen purge. We then used a mock circulatory system to test our modifications and make sure the LVAD continued to have appropriate flow rates. We then conducted training for staff and developed a disaster plan should the LVAD fail at any point. RESULTS: Once we felt comfortable with the modifications and had a plan developed should any problems arise, we then proceeded to treat our patient in the hyperbaric chamber. He successfully underwent 44 hyperbaric treatments for radiation cystitis without complications. CONCLUSION: This case is the second reported patient in the literature with an LVAD that was successfully treated in a multiplace hyperbaric chamber. As LVADs become increasingly popular to manage heart failure, more patients with these devices will present for hyperbaric treatments. With a few modifications, an LVAD patient can be safely and successfully treated in a hyperbaric chamber.

First Complete Genome Sequences of Zika Virus Isolated from Febrile Patient Sera in Ecuador.

Here, we present the complete genome sequences of two Zika virus (ZIKV) strains, EcEs062_16 and EcEs089_16, isolated from the sera of febrile patients in Esmeraldas City, in the northern coastal province of Esmeraldas, Ecuador, in April 2016. These are the first complete ZIKV genomes to be reported from Ecuador.

The potential application of a biomarker approach for the investigation of low-calorie sweetener exposure.

Low-calorie sweeteners (LCS) are commonly used as sugar substitutes in the diet to provide a desired sweet taste without increased energy intake. The number of LCS available on the market has increased considerably over the years and despite extensive evaluation of their safety prior to approval, debate continues around the effects of consumption on health. In Europe, Member States are obligated to monitor exposure to LCS and methods currently used tend to rely on self-reported dietary intake data alongside LCS concentrations in products. However, the acquisition of accurate data can be costly in terms of resources and time and are inherently imprecise. Although LCS are intensely sweet, they are chemically diverse and a limitation of many studies investigating the health effects of consumption is that they often fail to discern intakes of individual LCS. An approach which objectively assesses intakes of individual LCS would therefore allow robust investigations of their possible effects on health. Biomarker approaches have been utilised for the objective investigation of intakes of a range of dietary components and the feasibility of any such approach depends upon its validity as well as its applicability within the target population. This review aims to provide an overview of current understanding of LCS intake and explore the possibility of implementing a biomarker approach to enhance such understanding. Several commonly used LCS, once absorbed into the body, are excreted via the kidneys; therefore a urinary biomarker approach may be possible for the investigation of short-term exposure to these compounds.

The continued emergence of hantaviruses: isolation of a Seoul virus implicated in human disease, United Kingdom, October 2012.

Following a suspected case of hantavirus in a patientsuffering from acute kidney injury, rodents fromthe patient's property in Yorkshire and the Humber,United Kingdom (UK) were screened for hantaviruses.Hantavirus RNA was detected via RT-PCR in two Rattusnorvegicus. Complete sequencing and phylogeneticanalysis established the virus as a Seoul hantavirus,which we have provisionally designated as strainHumber. This is the first hantavirus isolated from wildrodents in the UK and confirms the presence of a pathogenicSeoul virus in Europe.

Sequencing and phylogenetic characterisation of a fatal Crimean - Congo haemorrhagic fever case imported into the United Kingdom, October 2012.

A patient with fever, and haemorrhagic symptoms was admitted to a hospital in Glasgow on 2 October 2012. Since he had returned from Afghanistan, serum samples were sent for diagnosis at the Rare and Imported Pathogens Laboratory, where a real-time reverse transcriptase-PCR diagnosis of Crimean ­ Congo haemorrhagic fever was made within 3 hrs after receipt of the sample. Hereafter the patient was transferred to a high-security infectious diseases unit in London but died on 6 October.

Methicillin-resistant Staphylococcus pseudintermedius isolated from canine pyoderma in North China.

AIMS: To determine the prevalence of carriage of methicillin-resistant Staphylococcus pseudintermedius (MRSP) among dogs with pyoderma from two small animal hospitals in North China during a 21-month period and to characterize these isolates. METHODS AND RESULTS: Swabs were taken from 260 dogs with pyoderma, and the staphylococcal species isolated and methicillin resistance were confirmed phenotypically and genotypically. The identified MRSP isolates were characterized by multilocus sequence typing (MLST), spa typing, staphylococcal cassette chromosome (SCC) mec typing, testing for susceptibility to nine antimicrobial agents and SmaI-digested pulsed-field gel electrophoresis. Thirty-three (12·7%) dogs were positive for MRSP. The most prevalent genotypes detected among MRSP were ST71(MLST)-t06(spa)-II-III(SCCmec) (n = 22, 66·7%), followed by ST5-t19 (n = 8, 24·2%), ST126-III(n = 2, 6·1%) and ST6-t02-V (3·0%). All MRSP isolates showed extended resistance to tested antimicrobial agents. Eight different SmaI patterns were observed in 21 typeable MRSP isolates. CONCLUSIONS: Clinical isolates of MSRP isolated from dogs in North China belonged to two major clonal lineages ST71 and ST5. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first report on MRSP from canine pyoderma in China. Further surveillance study is needed to gain more detailed data concerning this major clinical challenge in veterinary medicine.

Repeated therapeutic dosing selects macrolide-resistant Campylobacter spp. in a turkey facility.

AIMS: This study assessed the effects of the therapeutic use of Tylan® in a large-scale turkey production facility on the selection of macrolide-resistant Campylobacter. METHODS AND RESULTS: A flock of production turkeys (c. 30,000 birds) was followed from brooding to slaughter, and the effects of macrolide application was assessed in one half of the flock from finishing stage to final product and compared against the control barn where no macrolide was used. Overall, Campylobacter prevalence in turkeys was almost 100% by 4 weeks of age. When Campylobacter prevalence was assessed in relation to treatment, high levels of macrolide resistance were evident in this group following treatment, with Campylobacter coli becoming the dominant strain type. Over time, and in the absence of a selection agent, the population of resistant strains decreased suggesting that there was a fitness cost associated with macrolide resistance carriage and persistence. Macrolide resistance was detected in the control barn at a very low level (four isolates recovered during the study), suggesting that the creation or selection of macrolide-resistant Campylobacter was correlated with the treatment regime used. Molecular analysis of a selection of macrolide-resistant Campylobacter recovered was assessed using PCR, RFLP and sequence analysis of the 23S rRNA. The majority of isolates displaying high-level macrolide resistance (>256 µg ml(-1)) possessed an A2075G transition mutation in the 23S rRNA and the CmeABC efflux pump. CONCLUSIONS: These studies suggest that macrolide resistance can be promoted through the application of treatment during the grow-out phase and once established in a production facility has the potential to persist and be transferred to final product. SIGNIFICANCE AND IMPACT OF THE STUDY: The study highlights the prudent use of antimicrobials in treatment of disease in poultry. Of significance is the presence of macrolide-resistant Campylobacter in poultry production and finished product as a consequence of macrolide usage.

Aequorin variants with improved bioluminescence properties.

The photoprotein aequorin has been widely used as a bioluminescent label in immunoassays, for the determination of calcium concentrations in vivo, and as a reporter in cellular imaging. It is composed of apoaequorin (189 amino acid residues), the imidazopyrazine chromophore coelenterazine and molecular oxygen. The emission characteristics of aequorin can be changed by rational design of the protein to introduce mutations in its structure, as well as by substituting different coelenterazine analogues to yield semi-synthetic aequorins. Variants of aequorin were created by mutating residues His16, Met19, Tyr82, Trp86, Trp108, Phe113 and Tyr132. Forty-two aequorin mutants were prepared and combined with 10 different coelenterazine analogues in a search for proteins with different emission wavelengths, altered decay kinetics and improved stability. This spectral tuning strategy resulted in semi-synthetic photoprotein mutants with significantly altered bioluminescent properties.

First Report of the Oak Wilt Fungus, Ceratocystis fagacearum, in New York State.

Ceratocystis fagacearum (T.W. Bretz) J. Hunt is considered to be the most destructive vascular pathogen of oaks (Fagaceae: Quercus spp.) in the eastern, north-central, and south-central United States. (1,2,4). All red oak species (subgenus Quercus: section Lobatae) are highly susceptible to C. fagacearum, and infected trees typically die within 3 months of first symptom expression. However, members of the white oak group (subgenus Quercus: section Quercus) are moderately to highly resistant to C. fagacearum and rarely die from oak wilt (1,3). In early August of 2008, we received branch samples taken from wilting red oaks (Q. rubra L.) growing in a residential neighborhood in Scotia, NY (Schenectady County). The endoconidial state of the oak wilt fungus, Thielaviopsis quercina (B.W. Henry) A.E. Paulin, T.C. Harr. & McNew, was consistently isolated from the xylem in the branches. The cultures were identified based on hyphal and conidial morphology on acidified potato dextrose agar (aPDA) as well as sequences of the internal transcribed spacer (ITS) (GenBank Accession No. FJ347031) and large subunit (LSU) (GenBank Accession No. FJ347030) regions of nuclear ribosomal DNA (rDNA). The nucleotide identities for the ITS and LSU sequences were a precise match, 99 and 100%, to rDNA sequences (GenBank Accession Nos. AF043598 and AF222483, respectively) of other isolates of C. fagacearum. According to the homeowners at the site, 12 red oaks have died during the last 3 years, and each tree died within one growing season after oak wilt-like symptoms were noted. In a brief survey of nearby properties in late August of 2008, we found 12 additional trees that were either expressing crown symptoms of oak wilt (e.g., premature leaf casting, bronzing of leaf margins, and water-soaked leaves) or were standing dead and within close proximity (5 to 10 m) to symptomatic trees. Branch samples from four of the symptomatic trees revealed limited (spotted) or no vascular discoloration; however, C. fagacearum was isolated from each suspect tree on aPDA. Remnants of gray mycelial mats and associated pressure cushions were observed beneath the bark of one standing dead oak. The sweet fruit-like odor characteristic of the oak wilt fungus was immediately evident once the bark overlying the mats was removed. Prior to this discovery, the Susquehanna River in north-central Pennsylvania was considered to be the northeastern limit for oak wilt occurrence in the United States (2,4). To our knowledge, this is the first report of the fungus from New York and expands the known range of C. fagacearum to the northeast by at least 300 km, supporting the hypothesis that the range of this fungus continues to expand via animal vectors and/or human activities (2). An isolate of C. fagacearum from New York has been deposited at the Centraalbureau voor Schimmelcultures (CBS 123913). References: (1) D. N. Appel. Ann. Rev. Phytopathol. 33:103, 1995. (2) J. Juzwik et al. Ann. Rev. Phytopathol. 46:13, 2008. (3) W. L. MacDonald et al. European oaks-susceptible to oak wilt? Page 131 in: Shade Tree Wilt Diseases. C. L. Ash, ed. The American Phytopathological Society, St. Paul, MN, 2001. (4) USDA Forest Service. Oak Wilt Distribution. Northeast Area, State and Private Forestry, St. Paul, MN. Online publication, 2005.

Tracking epidemic Chikungunya virus into the Indian Ocean from East Africa.

The largest documented outbreak of Chikungunya virus (CHIKV) disease occurred in the Indian Ocean islands and India during 2004-2007. The magnitude of this outbreak led to speculation that a new variant of the virus had emerged that was either more virulent or more easily transmitted by mosquito vectors. To study this assertion, it is important to know the origin of the virus and how the particular strain circulating during the outbreak is related to other known strains. This study genetically characterized isolates of CHIKV obtained from Mombasa and Lamu Island, Kenya, during 2004, as well as strains from the 2005 outbreak recorded in Comoros. The results of these analyses demonstrated that the virus responsible for the epidemic that spread through the Indian Ocean originated in coastal Kenya during 2004 and that the closest known ancestors are members of the Central/East African clade. Genetic elements that may be responsible for the scope of the outbreak were also identified.

Spectral tuning of photoproteins by partnering site-directed mutagenesis strategies with the incorporation of chromophore analogs.

Aequorin and obelin are photoproteins whose calcium controlled bioluminescent light emission is used for labeling in assays, for the determination of calcium concentrations in vivo, and as a reporter in cellular imaging. Both of these photoproteins emit blue light from a 2-hydroperoxycoelenterazine chromophore, which is non-covalently bound in the hydrophobic core of the proteins. In an effort to produce aequorin and obelin variants with improved analytical properties, such as alternative emission colors and altered decay kinetics, seven mutants of aequorin and obelin were prepared and combined with 10 different coelenterazine analogs. These semi-synthetic photoprotein mutants exhibited shifts in bioluminescent properties when compared with wild-type proteins. The bioluminescent parameters determined for these semi-synthetic photoprotein mutants included specific activity, emission spectra and decay half-life time. This spectral tuning strategy resulted in semi-synthetic photoprotein mutants that had significantly altered bioluminescent properties. The largest emission maxima shift obtained was 44 nm, and the largest decay half-life difference was 23.91 s.

Characterization of antimicrobial susceptibility and virulence genes of Salmonella serovars collected at a commercial turkey processing plant.

AIMS: To determine the antimicrobial susceptibility profiles, distribution of class 1 integrons, virulence genes and genes encoding resistance to tetracycline (tetA, tetC, tetD and tetE) and streptomycin (strA, strB and aadA1) in Salmonella recovered from turkeys. METHODS AND RESULTS: The antimicrobial susceptibility of 80 isolates was determined using National Antimicrobial Resistance Monitoring System. The distribution of resistance genes, class 1 integrons and virulence genes was determined using PCR. Resistances to tetracycline (76 x 3%) and streptomycin (40%) were common. Sixty-two (77 x 5%) isolates displayed resistance against one or more antimicrobials and 33 were multi-drug resistant. tetA was detected in 72 x 5% of the isolates, while tetC, tetD and tetE were not detected. The strA and strB genes were detected in 73 x 8% of the isolates. Two isolates possessed class 1 integrons of 1 kb in size, containing the aadA1 gene conferring resistance to streptomycin and spectinomycin. Fourteen of the virulence genes were detected in over 80% of the isolates. CONCLUSIONS: This study shows that continuous use of tetracycline and streptomycin in poultry production selects for resistant strains. The Salmonella isolates recovered possess significant ability to cause human illness. SIGNIFICANCE AND IMPACT OF THE STUDY: Information from this study can be employed in guiding future strategies for the use of antimicrobials in poultry production.

Characterization of antimicrobial resistant Escherichia coli isolated from processed bison carcasses.

AIM: To determine the phenotypic and genotypic antimicrobial susceptibility profiles of Escherichia coli from bison carcasses. METHODS AND MATERIALS: The antimicrobial resistance of 138 E. coli isolates recovered from processed bison carcasses was determined by using the National Antimicrobial Resistance Monitoring System panels, polymerase chain reaction assays, plasmid analysis and conjugation studies. RESULTS: Resistance to 14 of the 16 antimicrobials was observed. Twenty-three (16.7%) isolates displayed resistance to at least one antimicrobial agent. The most prevalent resistances were to tetracycline (13.0%), sulfamethoxazole (7.9%) and streptomycin (5.8%). No resistance was observed to amikacin and ciprofloxacin. Further analysis of 23 antimicrobial-resistant E. coli isolates showed the presence of resistance genes corresponding to their phenotypic profiles. Results of conjugation studies carried out showed most isolates tested were able to transfer their resistance to recipients. CONCLUSION: This study indicated that multidrug-resistant E. coli isolates are present in bison. However, the resistance rate is lower than that reported in other meat species. SIGNIFICANCE AND IMPACT OF THE STUDY: The beneficial effects of antimicrobial-free feeding practice in bison may be promoting a reduction in the prevalence of antimicrobial resistance in commensal flora of bison.

Detection of Iss and Bor on the surface of Escherichia coli.

AIMS: To confirm the presence of Iss and Bor on the outer membrane of Escherichia coli using Western blots of outer membrane protein (OMP) preparations and fluorescence microscopy, and explore the use of fluorescence microscopy for the detection of avian pathogenic E. coli (APEC) and diagnosis of avian colibacillosis. METHODS AND RESULTS: Knockout mutants of iss and bor were created using a one-step recombination of target genes with PCR-generated antibiotic resistance cassettes. Anti-Iss monoclonal antibodies (Mabs) that cross-react with Bor protein were used to study the mutants relative to the wild-type organism. These Mabs were used as reagents to study OMP preparations of the mutants with Western blotting and intact E. coli cells with fluorescence microscopy. Iss and Bor were detected in Western blots of OMP preparations of the wild type. Also, Iss was detected on Deltabor mutants, and Bor was detected on Deltaiss mutants. Iss and Bor were also detected on the surface of the intact, wild-type cells and mutants using fluorescence microscopy. CONCLUSIONS: These results demonstrate that Bor and Iss are exposed on E. coli's outer membrane where they may be recognized by the host's immune system. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first report confirming Iss' location in the outer membrane of an E. coli isolate. Such surface exposure has implications for the use of these Mabs for APEC detection and colibacillosis control.