Antimicrobial-resistant Enterobacteriaceae recovered from companion animal and livestock environments.

Antimicrobial-resistant bacteria represent an important concern impacting both veterinary medicine and public health. The rising prevalence of extended-spectrum beta-lactamase (ESBL), AmpC beta-lactamase, carbapenemase (CRE) and fluoroquinolone-resistant Enterobacteriaceae continually decreases the efficiency of clinically important antibiotics. Moreover, the potential for zoonotic transmission of antibiotic-resistant enteric bacteria increases the risk to public health. Our objective was to estimate the prevalence of specific antibiotic-resistant bacteria on human contact surfaces in various animal environments. Environmental surface samples were collected from companion animal shelters, private equine facilities, dairy farms, livestock auction markets and livestock areas of county fairs using electrostatic cloths. Samples were screened for Enterobacteriaceae expressing AmpC, ESBL, CRE or fluoroquinolone resistance using selective media. Livestock auction markets and county fairs had higher levels of bacteria expressing both cephalosporin and fluoroquinolone resistance than did equine, dairy, and companion animal environments. Equine facilities harboured more bacteria expressing cephalosporin resistance than companion animal shelters, but less fluoroquinolone resistance. The regular use of extended-spectrum cephalosporins in livestock populations could account for the increased levels of cephalosporin resistance in livestock environments compared to companion animal and equine facilities. Human surfaces, as well as shared human and animal surfaces, were contaminated with resistant bacteria regardless of species environment. Detecting these bacteria on common human contact surfaces suggests that the environment can serve as a reservoir for the zoonotic transmission of antibiotic-resistant bacteria and resistance genes. Identifying interventions to lower the prevalence of antibiotic-resistant bacteria in animal environments will protect both animal and public health.

Gestation length and racing performance in 115 Thoroughbred foals with incomplete tarsal ossification.

BACKGROUND: Incomplete ossification of the cuboidal bones of the carpus and tarsus in foals has the potential for significant consequences including chronic lameness and decreased athletic ability. OBJECTIVES: To determine if the degree of ossification of the cuboidal bones is associated with gestational length and if the diagnosis of incomplete ossification is a predictor of performance in Thoroughbred racehorses. STUDY DESIGN: Retrospective cohort study. METHODS: The medical records of Thoroughbred foals less than 90 days of age from 1994 to 2011 were examined and records containing tarsal radiographs identified. Radiographs of the tarsus were examined for signs of incomplete ossification and those that were incompletely ossified graded on a scale of 1-4 using a modification of a previously reported index, with Grade 1 being the least ossified and Grade 4 being the most. Gestational length was determined by examining breeding records and foaling dates reported to the Jockey Club. Race records for 2- and 3-year-old affected foals and their maternal siblings were obtained and compared. RESULTS: Foals with Grades 1 and 2 ossification were usually premature (gestation length <325 days), but Grades 3 and 4 were not. Foals with Grades 2 and 3 ossification were significantly less likely to race than their maternal siblings and Grades 1, 2, 3 and 4 foals earned less money. MAIN LIMITATIONS: A larger sample size of foals with Grade 1 ossification would increase the power of the study. Foals radiographed at an older age may have had lower ossification scores if radiographed earlier. CONCLUSIONS: Incomplete ossification, especially Grades 1 and 2, is associated with a short gestation length. Foals with Grades 2 and 3 incomplete ossification were less likely to race and Grades 1, 2 and 3 earned around $30,000 less than their maternal siblings.

Genotypic and epidemiologic characterization of extended-spectrum cephalosporin resistant Salmonella enterica from US beef feedlots.

In the US, nontyphoidal Salmonellae are a common foodborne zoonotic pathogen causing gastroenteritis. Invasive Salmonella infections caused by extended-spectrum cephalosporin resistant (ESCR) phenotypes are more likely to result in treatment failure and adverse health outcomes, especially in severe pediatric Salmonella infections where the extended-spectrum ß-lactams are the therapy of choice. To examine the genetic and epidemiologic characteristics of ESCR Salmonellae which may enter the food chain, we characterized 44 ceftiofur-resistant Salmonella isolates from the National Animal Health Monitoring System (NAHMS) 2011 beef cattle feedlot health and management study. As part of the NAHMS Feedlot 2011 study, 5050 individual fecal samples from 68 large (1000+ head capacity) feedlots were cultured for Salmonella spp. The resulting 460 positive samples yielded 571 Salmonella isolates with 44 (8%) expressing an AmpC ß-lactamase phenotype. These phenotypic blaCMY-2Salmonella isolates represented 8 serotypes, most commonly S. Newport (n=14, 32%), S. Typhimurium (n=13, 30%), and S. Reading (n=5, 11%), followed by S. Dublin, S. Infantis, S. Montevideo, S. Rough O:i;v:1;7, and S. Uganda. Carriage of the blaCMY-2 gene was confirmed for all isolates expressing an AmpC ß-lactamase phenotype by PCR. Additionally, all 44 isolates were shown to carry the blaCMY-2 gene on a large IncA/C plasmid, a gene/plasmid combination which has been previously reported in multiple species. Other plasmids, including IncN, FIC, and FIIA, were also detected in some isolates. Cattle fed chlortetracycline were less likely to be positive for a blaCMY-2Salmonella isolate in their enteric flora compared to those not receiving chlortetracycline during the feeding period. Carriage of blaCMY-2 was more prevalent in Salmonella isolates originating from lighter weight cattle, cattle fed tylosin and dairy breeds. Our characterization of the NAHMS Feedlot 2011 study Salmonella isolates with ESCR phenotype shows that while other cephalosporin resistance mechanisms have been reported in US cattle, specific serotypes harboring blaCMY-2 on IncA/C plasmids may be the dominant resistance genotype.

Prevalence of AmpC- and Extended-Spectrum ß-Lactamase-Harbouring Enterobacteriaceae in Faecal Flora of a Healthy Domestic Canine Population.

In order to estimate the prevalence of AmpC- and ESBL ß-lactamase-producing Enterobacteriaceae in the faecal flora of a healthy domestic canine population, faecal samples were obtained from healthy dogs receiving routine parasitology screening at the Ohio State University Veterinary Medical Center, between January 2013 and April 2013. Samples were screened for the presence of AmpC and ESBL ß-lactamase phenotypes, and the clinically important genotypes, blaCMY and blaCTX-M , were confirmed via conventional PCR. Minimum inhibitory concentrations were determined for isolates and plasmids were characterized. Two hundred and twelve canine faecal samples were screened, of which 30 harboured isolates carrying the AmpC blaCMY , representing 14.2% of the population (95% CI: 9.4-18.9%). Nine samples harboured isolates that carried the ESBL blaCTX-M , representing 4.2% of the population (95% CI: 1.5-7.0%). Isolates containing blaCMY harboured multiple plasmid replicon types, while isolates containing blaCTX-M harboured few plasmid replicon types. Our results suggest that domestic dogs may serve as a reservoir for extended-spectrum cephalosporin resistance genes for other domestic animal populations as well as for their human companions. This represents a potential veterinary and public health risk that warrants further investigation and continued surveillance to ascertain the nature and extent of the risk. The high level of diversity of plasmid content among isolates harbouring blaCMY suggests broader dissemination relative to blaCTX-M isolates.

Rhodium and silicon system: II. Rhodium silicide formation.

Detailed characterizations of rhodium/silicon films prepared by co-deposition using magnetron sputtering have been carried out on silicon substrates at room temperature up to 900 degrees C. The properties of the films were investigated using XPS/UPS, XRD, SIMS, SEM and AFM techniques. It should be emphasized that XPS/UPS measurements are carried out without breaking the vacuum to avoid any contamination of the film. Up to 500 degrees C an interdiffusion between the oxidized silicon wafer and the deposited Rh/Si film occurred leading to hole formation in the entire film at 900 degrees C. Diffraction patterns for the compounds Rh(2)Si, Rh(5)Si(3), RhSi and Rh(3)Si(4) were measured. Upon annealing the covalent character is increased and for the samples forming the compound RhSi the valence band structure is markedly changed. Depth profiling (XPS and SIMS) reveals a stable composition in the bulk of the film. For these measurements the silicon-rich alloy in the interfacial layer is probably an effect of sputtering, by implanting the Rh atoms into the silicon substrate. A previously reported negative shift for the compound Rh(5)Si(3) could be connected to the sample preparation, as sputtering of the surface is reducing the silicon content and inducing a glassy state. For the first phase Rh(2)Si formed on the rhodium-rich side the shift in binding energy is unclear, for all the other compounds encountered in this work a positive shift relative to pure rhodium was found.

Optical characterization of alignment and effective refractive index in carbon nanotube films.

We have characterized the thickness and effective refractive index of carbon nanotube forests by fitting reflectance measurements in the visible and near infrared ranges. The measurements were performed with polarized light. An effective medium layer consisting of a mixture of graphite and air was used to simulate the nanotube film. The proposed model accurately described the behaviour of the reflected s-polarized component (Rs), which allowed for the precise determination of the thickness and porosity of the films, in very good agreement with SEM measurements of film thickness. The p-polarized component (Rp), on the other hand, could not be described in terms of the developed model. In badly aligned samples, where there is a mixture of Rs and Rp behaviour, the model fails to fit the Rs component as well. This effect can therefore be taken as an indirect indication of lack of alignment in the samples.

Evaluation of the interface between one-bottle bonding agents and dentin by cryopreparation and low-temperature scanning electron microscopy (LTSEM). A pilot study on perfused dentinal samples.

OBJECTIVES: Dentin bonding samples are generally observed in the high vacuum chamber of an electron microscope only after completion of the bonding procedure fixation, dehydration and drying of the specimens. The purpose of this study was to observe the effect of each step in the application of various one-bottle dentin bonding agents (DBA) using cryopreparation followed by low-temperature scanning electron microscopy (LTSEM). METHODS: Prime&Bond 2.1 (P&B 2.1), Coltène Experimental (Exp.) and Syntac Single Component (Syntac SC) were applied onto perfused dentin of extracted, human third permanent molar teeth. After acid-etching, rinsing and drying, and following the application of the respective resins, the teeth were fractured at random and plunge-frozen in liquid nitrogen before examination under LTSEM. As this method preserves the state of the treated surface at the moment when it is frozen, each step of the application of the DBA can be observed as if it were a 'real-time' procedure. RESULTS: Acid-etching the dentin resulted in the removal of the smear layer for all materials tested. Those one-bottle DBAs which recommend the application of two consecutive resin layers (P&B 2.1 and Syntac SC) showed incomplete saturation of the dentinal surface after application of the first layer. With Syntac SC the incomplete saturation was more pronounced than with P&B 2.1. The use of Exp. resulted in a much more homogeneous coverage of the dentin despite only one layer of resin having been applied. CONCLUSION: It is concluded that cryo-preparation followed by LTSEM appears to be a method which allows each stage of the application of DBA to be evaluated.

Trichorhinophalangeal syndrome type III.

Trichorhinophalangeal syndrome (TRPS) type III is a newly defined clinical entity. This symptom complex is inherited as an autosomal dominant trait and clinically characterized by growth retardation, craniofacial abnormalities, severe brachydactyly and sparse hair. In addition, absence of mental retardation and cartilaginous exostoses are required for the diagnosis of TRPS III. To further delineate this newly recognized entity, we report on a patient from a Turkish family segregating TRPS III in 7 family members. The patient had a very short stature (147 cm, < 3rd standard deviation), a thin upper lip and a prominent lower lip, a pear-shaped nose, stubby fingers and toes with cone-shaped epiphyses and sparse scalp hair. Scanning electron microscopy findings and results of energy-dispersive X-ray microanalysis are presented in such a patient for the first time.

Oxidation of midazolam and triazolam by human liver cytochrome P450IIIA4.

The metabolism of midazolam and triazolam to their 1'-hydroxy and 4-hydroxy metabolites was studied in microsomes of 15 human livers. The formation of both metabolites was inhibited by more than 90% by an antiserum directed against a pregnenolone 16 alpha-carbonitrile-inducible cytochrome P450 (P450PCN1) of rat liver. Moreover, midazolam hydroxylase activity was immunoprecipitated from solubilized human microsomes with polyclonal antibodies against rat P450PCN1 and the closely related human isozyme P450NF. A close correlation was observed between the amount of protein detected in immunoblots with these antibodies and the midazolam or triazolam hydrxylase activity. The formation of both metabolites of midazolam was inhibited by triacetyloleandomycin, a known inhibitor of cytochromes P450 of the IIIA family. Direct evidence that P450IIIA4 catalyzes the metabolism of midazolam was provided through the use of cDNA-directed expression. Monkey COS cells transfected with human P450PCN1 cDNA were able to catalyze both the 1'- and the 4-hydroxylation of midazolam. We conclude that the metabolism of midazolam and triazolam in human liver is predominantly mediated by cytochrome P450IIIA4. Two of 15 human livers expressed a second immunoreactive microsomal protein of higher apparent Mr and were more active in midazolam 1'-hydroxylation. Our data also provide evidence that the marked interindividual variation in the response to these widely used benzodiazepine drugs is due to variable hepatic metabolism.

High-performance liquid chromatographic assays for bufuralol 1'-hydroxylase, debrisoquine 4-hydroxylase, and dextromethorphan O-demethylase in microsomes and purified cytochrome P-450 isozymes of human liver.

Bufuralol, debrisoquine, and dextromethorphan are three prototype substrates of the common genetic deficiency of oxidative drug metabolism in man known as debrisoquine/sparteine-type polymorphism. We describe assays for the in vitro metabolism of (+)- and (-)-bufuralol, debrisoquine, and dextromethorphan in human liver microsomes and reconstituted purified cytochrome P-450 isozymes. These assays combine nonextractive sample preparation by precipitation of protein with perchloric acid with reversed-phase inorganic ion-pair HPLC and fluorescence detection. The minimal detectable levels of the major metabolites formed are 1'-hydroxybufuralol, 0.1 ng/ml; 4-hydroxydebrisoquine, 0.8 ng/ml; and dextrorphan, 0.1 ng/ml. Formation of these metabolites is linear for at least 45 min and between 1 and 100 micrograms of microsomal protein. Comparative kinetic analysis of the three monooxygenase reactions in human liver microsomes revealed an apparent biphasicity of (+)- and (-)-bufuralol 1'-hydroxylation and dextromethorphan O-demethylation but monophasic formation of 4-hydroxydebrisoquine in the substrate concentration range (less than 1 mM) studied. These data, in combination with those obtained by purified human cytochrome P-450 isozymes indicate the involvement of the same enzyme in the metabolism of all three substrates investigated. However, additional and distinct activities contribute to the metabolism of (+)- and (-)-bufuralol and dextromethorphan.