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1.
Foodborne Pathog Dis ; 15(6): 377-385, 2018 06.
Article in English | MEDLINE | ID: mdl-29638171

ABSTRACT

Campylobacter spp. can be pathogenic to humans and often harbor antimicrobial resistance genes. Data on resistance in relation to fluoroquinolone use in beef cattle are scarce. This cross-sectional study of preharvest cattle evaluated Campylobacter prevalence and susceptibility to nalidixic acid and ciprofloxacin in feedlots that previously administered a fluoroquinolone as primary treatment for bovine respiratory disease. Twenty fresh fecal samples were collected from each of 10 pens, in each of five feedlots, 1-2 weeks before harvest. Feces were cultured for Campylobacter using selective enrichment and isolation methods. Genus and species were confirmed via PCR. Minimum inhibitory concentrations (MICs) of ciprofloxacin and nalidixic acid were determined using a micro-broth dilution method and human breakpoints. Antimicrobial use within each pen was recorded. Data were analyzed using generalized linear mixed-models (prevalence) and survival analysis (MICs). Overall, sample-level prevalence of Campylobacter was 27.2% (272/1000) and differed significantly among feedlots (p < 0.01). Campylobacter coli was the most common species (55.1%; 150/272), followed by Campylobacter hyointestinalis (42.6%; 116/272). Within-pen prevalence was not significantly associated with the number of fluoroquinolone treatments, sex, body weight, or metaphylaxis use, but was associated with the number of days cattle were in the feedlot (p = 0.03). The MICs for the majority of Campylobacter isolates were above the breakpoints for nalidixic acid (68.4%; 175/256) and for ciprofloxacin (65.6%; 168/256). Distributions of MICs for nalidixic acid (p ≤ 0.01) and ciprofloxacin (p ≤ 0.05) were significantly different among feedlots, and by Campylobacter species. However, fluoroquinolone treatments, sex, body weight, days on feed, and metaphylaxis were not significantly associated with MIC distributions within pens. We found no evidence that the number of fluoroquinolone treatments within feedlot pens significantly affected the within-pen fecal prevalence or quinolone susceptibilies of Campylobacter in feedlots that used a fluoroquinolone as primary treatment for bovine respiratory disease.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Campylobacter Infections/veterinary , Campylobacter/drug effects , Cattle Diseases/epidemiology , Enrofloxacin/therapeutic use , Quinolones/pharmacology , Animals , Anti-Bacterial Agents/pharmacology , Campylobacter/isolation & purification , Campylobacter Infections/drug therapy , Campylobacter Infections/epidemiology , Campylobacter Infections/microbiology , Cattle , Cattle Diseases/drug therapy , Cattle Diseases/microbiology , Cross-Sectional Studies , Drug Resistance, Bacterial , Kansas/epidemiology , Prevalence , Texas/epidemiology
2.
Foodborne Pathog Dis ; 14(10): 600-607, 2017 10.
Article in English | MEDLINE | ID: mdl-28768141

ABSTRACT

The study objective was to determine effects of fluoroquinolone metaphylaxis on fecal prevalence of Salmonella and Campylobacter and fecal prevalence of quinolone-resistant Salmonella and Campylobacter in feedlot cattle. On Day 0, cattle (n = 288) at risk for bovine respiratory disease (BRD) were randomly assigned to either a nontreated control pen (12 pens) or a fluoroquinolone-treated (enrofloxacin; Baytril® 100) pen (12 pens). Rectal fecal samples were collected from cattle on days 0, 7, 14, 21, and 28. Feces were cultured for Salmonella enterica and Campylobacter spp. using enrichment and selective isolation methods, and confirmed by serology and PCR. Susceptibilities to nalidixic acid and ciprofloxacin were determined using microbroth dilution methods. Data analyses were performed using linear mixed models. Overall, Salmonella sp. and Campylobacter spp. were recovered from 10.2% (139/1,364) and 12.4% (170/1,364) of the fecal samples, respectively. Campylobacter species included hyointestinalis, jejuni, and coli. Neither Salmonella sp. nor Campylobacter spp. prevalence was significantly impacted by fluoroquinolone treatment (p = 0.80, p = 0.61, respectively). However, Salmonella prevalence differed between study weeks (p < 0.01) with prevalence decreasing over time. Before treatment, 98.9% (91/92) of Salmonella isolates were susceptible to nalidixic acid and ciprofloxacin. All Salmonella recovered posttreatment (n = 43) were susceptible to both antimicrobials. The majority of Campylobacter spp. recovered before treatment were resistant to nalidixic acid (23/35; 65.7%) and ciprofloxacin (21/35; 60.0%). There was no significant treatment by week interaction (p = 0.85) or treatment effects (p = 0.61) on the posttreatment prevalence of Campylobacter resistance. There was, however, a significant week effect (p = 0.05), with Campylobacter resistance prevalence decreasing over time. In this 28-day study, we found no evidence that a fluoroquinolone used for metaphylaxis significantly impacts fecal prevalence of Salmonella sp. or Campylobacter spp. or the fecal prevalence of nalidixic acid or ciprofloxacin resistance.


Subject(s)
Campylobacter Infections/veterinary , Campylobacter/drug effects , Cattle Diseases/epidemiology , Foodborne Diseases/prevention & control , Salmonella Infections, Animal/epidemiology , Salmonella enterica/drug effects , Animals , Anti-Infective Agents/pharmacology , Campylobacter/isolation & purification , Campylobacter Infections/epidemiology , Campylobacter Infections/microbiology , Cattle , Cattle Diseases/microbiology , Drug Resistance, Bacterial , Feces/microbiology , Fluoroquinolones/pharmacology , Foodborne Diseases/microbiology , Humans , Microbial Sensitivity Tests , Prevalence , Quinolones/pharmacology , Random Allocation , Salmonella Infections, Animal/microbiology , Salmonella enterica/isolation & purification
3.
BMC Genomics ; 18(1): 78, 2017 01 13.
Article in English | MEDLINE | ID: mdl-28086803

ABSTRACT

BACKGROUND: 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is the most potent congener of the dioxin class of environmental contaminants. Exposure to TCDD causes a wide range of toxic outcomes, ranging from chloracne to acute lethality. The severity of toxicity is highly dependent on the aryl hydrocarbon receptor (AHR). Binding of TCDD to the AHR leads to changes in transcription of numerous genes. Studies evaluating the transcriptional changes brought on by TCDD may provide valuable insight into the role of the AHR in human health and disease. We therefore compiled a collection of transcriptomic datasets that can be used to aid the scientific community in better understanding the transcriptional effects of ligand-activated AHR. RESULTS: Specifically, we have created a datasets package - TCDD.Transcriptomics - for the R statistical environment, consisting of 63 unique experiments comprising 377 samples, including various combinations of 3 species (human derived cell lines, mouse and rat), 4 tissue types (liver, kidney, white adipose tissue and hypothalamus) and a wide range of TCDD exposure times and doses. These datasets have been fully standardized using consistent preprocessing and annotation packages (available as of September 14, 2015). To demonstrate the utility of this R package, a subset of "AHR-core" genes were evaluated across the included datasets. Ahrr, Nqo1 and members of the Cyp family were significantly induced following exposure to TCDD across the studies as expected while Aldh3a1 was induced specifically in rat liver. Inmt was altered only in liver tissue and primarily by rat-AHR. CONCLUSIONS: Analysis of the "AHR-core" genes demonstrates a continued need for studies surrounding the impact of AHR-activity on the transcriptome; genes believed to be consistently regulated by ligand-activated AHR show surprisingly little overlap across species and tissues. Until now, a comprehensive assessment of the transcriptome across these studies was challenging due to differences in array platforms, processing methods and annotation versions. We believe that this package, which is freely available for download ( http://labs.oicr.on.ca/boutros-lab/tcdd-transcriptomics ) will prove to be a highly beneficial resource to the scientific community evaluating the effects of TCDD exposure as well as the variety of functions of the AHR.


Subject(s)
Environmental Pollutants/pharmacology , Gene Expression Profiling , Gene Expression Regulation/drug effects , Polychlorinated Dibenzodioxins/pharmacology , Transcriptome , Animals , Cell Line , Computational Biology/methods , Female , Gene Expression Profiling/methods , Humans , Male , Mice , Rats , Software , Web Browser
4.
Arch Toxicol ; 91(1): 325-338, 2017 Jan.
Article in English | MEDLINE | ID: mdl-27136898

ABSTRACT

2,3,7,8 Tetrachlorodibenzo-p-dioxin (TCDD) is an aromatic, long-lived environmental contaminant. While the pathogenesis of TCDD-induced toxicity is poorly understood, it has been shown that the aryl hydrocarbon receptor (AHR) is required. However, the specific transcriptomic changes that lead to toxic outcomes have not yet been identified. We previously identified a panel of 33 genes that respond to TCDD treatment in two TCDD-sensitive rodent species. To identify genes involved in the onset of hepatic toxicity, we explored 25 of these in-depth using liver from two rat strains: the TCDD-resistant Han/Wistar (H/W) and the TCDD-sensitive Long-Evans (L-E). Time course and dose-response analyses of mRNA abundance following TCDD insult indicate that eight genes are similarly regulated in livers of both strains of rat, suggesting that they are not central to the severe L-E-specific TCDD-induced toxicities. The remaining 17 genes exhibited various divergent mRNA abundances between L-E and H/W strains after TCDD treatment. Several genes displayed a biphasic response where the initial response to TCDD treatment was followed by a secondary response, usually of larger magnitude in L-E liver. This secondary response was most often an exaggeration of the original TCDD-induced response. Only cytochrome b5 type A (microsomal) (Cyb5a) had equivalent TCDD sensitivity to the prototypic AHR-responsive cytochrome P450, family 1, subfamily a, polypeptide 1 (Cyp1a1), while six genes were less sensitive. Four genes showed an early inter-strain difference that was sustained throughout most of the time course (atypical chemokine receptor 3 (Ackr3), collagen, type XVIII, alpha 1 (Col18a1), Cyb5a and glutamate dehydrogenase 1 (Glud1)), and of those genes examined in this study, are most likely to represent genes involved in the pathogenesis of TCDD-induced hepatotoxicity in L-E rats.


Subject(s)
Carcinogens, Environmental/toxicity , Chemical and Drug Induced Liver Injury/metabolism , Gene Expression Regulation/drug effects , Liver/drug effects , Polychlorinated Dibenzodioxins/toxicity , RNA, Messenger/metabolism , Animals , Animals, Outbred Strains , Carcinogens, Environmental/administration & dosage , Chemical and Drug Induced Liver Injury/enzymology , Collagen Type VIII/agonists , Collagen Type VIII/antagonists & inhibitors , Collagen Type VIII/genetics , Collagen Type VIII/metabolism , Cytochromes b5/antagonists & inhibitors , Cytochromes b5/chemistry , Cytochromes b5/genetics , Cytochromes b5/metabolism , Dose-Response Relationship, Drug , Drug Resistance , Female , Gene Expression Profiling , Glutamate Dehydrogenase , Kinetics , Liver/enzymology , Liver/metabolism , Male , Mice, Inbred C57BL , Polychlorinated Dibenzodioxins/administration & dosage , Rats, Long-Evans , Receptors, CXCR/agonists , Receptors, CXCR/antagonists & inhibitors , Receptors, CXCR/genetics , Receptors, CXCR/metabolism , Receptors, Glutamate/chemistry , Receptors, Glutamate/genetics , Receptors, Glutamate/metabolism
5.
Foodborne Pathog Dis ; 13(6): 303-8, 2016 06.
Article in English | MEDLINE | ID: mdl-26990393

ABSTRACT

Salmonella is an important foodborne pathogen and antimicrobial resistance can be a human health concern. The objectives of this cross-sectional study were to (1) determine the prevalence and quinolone susceptibility of Salmonella in feces of preharvest commercial feedlot cattle and (2) determine if the prevalence and susceptibility of Salmonella isolates were associated with previous fluoroquinolone use within pens. Five feedlots in western Kansas and Texas were selected based on their use of a commercially licensed fluoroquinolone for initial treatment of bovine respiratory disease (BRD). Twenty pen floor fecal samples were collected from each of 10 pens from each feedlot during early summer of 2012. Salmonella isolation was performed and microbroth dilution was used to determine susceptibility of isolates to nalidixic acid and ciprofloxacin. Prior antimicrobial treatment data were retrieved from feedlots' operational data. Generalized linear mixed models were used to assess associations between Salmonella prevalence and the number of fluoroquinolone treatments within pens while taking into consideration cattle demographic and management factors, as well as the hierarchical structure of the data. Overall, cumulative fecal prevalence of Salmonella was 38.0% (380/1000), but prevalence varied significantly (p < 0.01) among the five feedlots: 0.5% (1/200), 17.5% (35/200), 37.0% (74/200), 58.5% (117/200), and 76.5% (153/200). Salmonella serogroups included C1 (49.3%), E (36.4%), C2 (13.8%), and D (0.6%). There was no significant association (p = 0.52) between Salmonella prevalence and the frequency of fluoroquinolone treatments within a pen. All Salmonella isolates (n = 380) were susceptible to ciprofloxacin, while one isolate exceeded the human breakpoint (≥32 µg/mL) for nalidixic acid. In conclusion, Salmonella fecal prevalence in preharvest cattle was highly variable among feedlots. Nearly all Salmonella isolates were susceptible to quinolones, despite the fact that a fluoroquinolone was used as the primary therapeutic antimicrobial to treat BRD in these feedlot populations.


Subject(s)
Cattle Diseases/microbiology , Fluoroquinolones/pharmacology , Respiratory Tract Infections/veterinary , Salmonella Infections, Animal/microbiology , Salmonella/drug effects , Abattoirs , Animal Husbandry , Animals , Cattle , Cattle Diseases/drug therapy , Cattle Diseases/epidemiology , Cross-Sectional Studies , Drug Resistance, Bacterial , Feces/microbiology , Female , Fluoroquinolones/therapeutic use , Food Microbiology , Kansas/epidemiology , Male , Meat , Microbial Sensitivity Tests/veterinary , Prevalence , Respiratory Tract Infections/drug therapy , Salmonella/isolation & purification , Salmonella Infections, Animal/drug therapy , Salmonella Infections, Animal/epidemiology , Texas/epidemiology
6.
Toxicol Appl Pharmacol ; 274(3): 445-54, 2014 Feb 01.
Article in English | MEDLINE | ID: mdl-24355419

ABSTRACT

Despite several decades of research, the complete mechanism by which 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and other xenobiotic agonists of the aryl hydrocarbon receptor (AHR) cause toxicity remains unclear. While it has been shown that the AHR is required for all major manifestations of toxicity, the specific downstream changes involved in the development of toxic phenotypes remain unknown. Here we examine a panel of 13 genes that are AHR-regulated in many species and tissues. We profiled their hepatic mRNA abundances in two rat strains with very different sensitivities to TCDD: the TCDD-sensitive Long-Evans (Turku/AB; L-E) and the TCDD-resistant Han/Wistar (Kuopio; H/W). We evaluated doses ranging from 0 to 3000µg/kg at 19h after TCDD exposure and time points ranging from 1.5 to 384h after exposure to 100µg/kg TCDD. Twelve of 13 genes responded to TCDD in at least one strain, and seven of these showed statistically significant inter-strain differences in the time course analysis (Aldh3a1, Cyp1a2, Cyp1b1, Cyp2a1, Fmo1, Nfe2l2 and Nqo1). Cyp2s1 did not respond to TCDD in either rat strain. Five genes exhibited biphasic responses to TCDD insult (Ahrr, Aldh3a1, Cyp1b1, Nfe2l2 and Nqo1), suggesting a secondary event, such as association with additional transcriptional modulators. Of the 12 genes that responded to TCDD during the dose-response analysis, none had an ED50 equivalent to that of Cyp1a1, the most sensitive gene in this study, while nine genes responded to doses at least 10-100 fold higher, in at least one strain (Ahrr (L-E), Aldh3a1 (both), Cyp1a2 (both), Cyp1b1 (both), Cyp2a1 (L-E), Inmt (both), Nfe2l2 (L-E), Nqo1 (L-E) and Tiparp (both)). These data shed new light on the association of the AHR target genes with TCDD toxicity, and in particular the seven genes exhibiting strain-specific differences represent strong candidate mediators of Type-II toxicities.


Subject(s)
Gene Expression Regulation/drug effects , Liver/drug effects , Polychlorinated Dibenzodioxins/toxicity , Receptors, Aryl Hydrocarbon/metabolism , Animals , Aryl Hydrocarbon Hydroxylases/genetics , Aryl Hydrocarbon Hydroxylases/metabolism , Cytochrome P-450 CYP1A2 , Cytochrome P-450 CYP1B1 , Cytochrome P450 Family 2 , Cytochromes/genetics , Cytochromes/metabolism , Dose-Response Relationship, Drug , Liver/metabolism , Male , NAD(P)H Dehydrogenase (Quinone)/genetics , NAD(P)H Dehydrogenase (Quinone)/metabolism , NF-E2-Related Factor 2/genetics , NF-E2-Related Factor 2/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Rats, Long-Evans , Rats, Wistar , Steroid Hydroxylases/genetics , Steroid Hydroxylases/metabolism , Xenobiotics/toxicity
7.
RNA ; 19(1): 51-62, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23169800

ABSTRACT

Profiling of mRNA abundances with high-throughput platforms such as microarrays and RNA-seq has become an important tool in both basic and biomedical research. However, these platforms remain prone to systematic errors and have challenges in clinical and industrial applications. As a result, it is standard practice to validate a subset of key results using alternate technologies. Similarly, clinical and industrial applications typically involve transitions from a high-throughput discovery platform to medium-throughput validation ones. These medium-throughput validation platforms have high technical reproducibility and reduced sample input needs, and low sensitivity to sample quality (e.g., for processing FFPE specimens). Unfortunately, while medium-throughput platforms have proliferated, there are no comprehensive comparisons of them. Here we fill that gap by comparing two key medium-throughput platforms--NanoString's nCounter Analysis System and ABI's OpenArray System--to gold-standard quantitative real-time RT-PCR. We quantified 38 genes and positive and negative controls in 165 samples. Signal:noise ratios, correlations, dynamic range, and detection accuracy were compared across platforms. All three measurement technologies showed good concordance, but with divergent price/time/sensitivity trade-offs. This study provides the first detailed comparison of medium-throughput RNA quantification platforms and provides a template and a standard data set for the evaluation of additional technologies.


Subject(s)
High-Throughput Nucleotide Sequencing/methods , RNA, Messenger/analysis , Real-Time Polymerase Chain Reaction/methods , Animals , Gene Expression Profiling/methods , High-Throughput Nucleotide Sequencing/economics , High-Throughput Nucleotide Sequencing/instrumentation , Male , Oligonucleotide Array Sequence Analysis/methods , Polychlorinated Dibenzodioxins/toxicity , RNA, Messenger/drug effects , Rats , Rats, Long-Evans , Rats, Wistar , Real-Time Polymerase Chain Reaction/economics , Real-Time Polymerase Chain Reaction/instrumentation , Reproducibility of Results , Signal-To-Noise Ratio
8.
Toxicol Appl Pharmacol ; 251(2): 119-29, 2011 Mar 01.
Article in English | MEDLINE | ID: mdl-21215274

ABSTRACT

The dioxin congener 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) causes a wide range of toxic effects in rodent species, all of which are mediated by a ligand-dependent transcription-factor, the aryl hydrocarbon receptor (AHR). The Han/Wistar (Kuopio) (H/W) strain shows exceptional resistance to many TCDD-induced toxicities; the LD50 of > 9600 µg/kg for H/W rats is higher than for any other wild-type mammal known. We previously showed that this resistance primarily results from H/W rats expressing a variant AHR isoform that has a substantial portion of the AHR transactivation domain deleted. Despite this large deletion, H/W rats are not entirely refractory to the effects of TCDD; the variant AHR in these animals remains fully competent to up-regulate well-known dioxin-inducible genes. TCDD-sensitive (Long-Evans, L-E) and resistant (H/W) rats were treated with either corn-oil (with or without feed-restriction) or 100 µg/kg TCDD for either four or ten days. Hepatic transcriptional profiling was done using microarrays, and was validated by RT-PCR analysis of 41 genes. A core set of genes was altered in both strains at all time points tested, including CYP1A1, CYP1A2, CYP1B1, Nqo1, Aldh3a1, Tiparp, Exoc3, and Inmt. Outside this core, the strains differed significantly in the breadth of response: three-fold more genes were altered in L-E than H/W rats. At ten days almost all expressed genes were dysregulated in L-E rats, likely reflecting emerging toxic responses. Far fewer genes were affected by feed-restriction, suggesting that only a minority of the TCDD-induced changes are secondary to the wasting syndrome.


Subject(s)
Dioxins/toxicity , Drug Resistance, Multiple/drug effects , Environmental Pollutants/toxicity , Gene Expression Profiling , Liver/drug effects , Polychlorinated Dibenzodioxins/toxicity , Animals , Dose-Response Relationship, Drug , Drug Resistance, Multiple/physiology , Gene Expression Profiling/methods , Liver/physiology , Male , Rats , Rats, Long-Evans , Rats, Wistar , Species Specificity
9.
Chem Commun (Camb) ; (29): 3679-81, 2005 Aug 07.
Article in English | MEDLINE | ID: mdl-16027909

ABSTRACT

Readily prepared imidazole-based boronium ions form stable, hydrophobic, room-temperature ionic liquids (RTIL) with unique electronic and spectroscopic characteristics.

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