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Frontiers in Microbiology 2014Antibiotics are used in livestock and poultry production to treat and prevent disease as well as to promote animal growth. Carbadox is an in-feed antibiotic that is...
Antibiotics are used in livestock and poultry production to treat and prevent disease as well as to promote animal growth. Carbadox is an in-feed antibiotic that is widely used in swine production to prevent dysentery and to improve feed efficiency. The goal of this study was to characterize the effects of carbadox and its withdrawal on the swine gut microbiota. Six pigs (initially 3-weeks old) received feed containing carbadox and six received unamended feed. After 3-weeks of continuous carbadox administration, all pigs were switched to a maintenance diet without carbadox. DNA was extracted from feces (n = 142) taken before, during, and following (6-week withdrawal) carbadox treatment. Phylotype analysis using 16S rRNA sequences showed the gradual development of the non-medicated swine gut microbiota over the 8-week study, and that the carbadox-treated pigs had significant differences in bacterial membership relative to non-medicated pigs. Enumeration of fecal Escherichia coli showed that a diet change concurrent with carbadox withdrawal was associated with an increase in the E. coli in the non-medicated pigs, suggesting that carbadox pre-treatment prevented an increase of E. coli populations. In-feed carbadox caused striking effects within 4 days of administration, with significant alterations in both community structure and bacterial membership, notably a large relative increase in Prevotella populations in medicated pigs. Digital PCR was used to show that the absolute abundance of Prevotella was unchanged between the medicated and non-medicated pigs despite the relative increase shown in the phylotype analysis. Carbadox therefore caused a decrease in the abundance of other gut bacteria but did not affect the absolute abundance of Prevotella. The pending regulation on antibiotics used in animal production underscores the importance of understanding how they modulate the microbiota and impact animal health, which will inform the search for antibiotic alternatives.
PubMed: 24959163
DOI: 10.3389/fmicb.2014.00276 -
MBio Aug 2017Carbadox is a quinoxaline-di--oxide antibiotic fed to over 40% of young pigs in the United States that has been shown to induce phage DNA transduction ; however, the...
Carbadox is a quinoxaline-di--oxide antibiotic fed to over 40% of young pigs in the United States that has been shown to induce phage DNA transduction ; however, the effects of carbadox on swine microbiome functions are poorly understood. We investigated the longitudinal effects of carbadox on swine gut microbial gene expression (fecal metatranscriptome) and phage population dynamics (fecal dsDNA viromes). Microbial metagenome, transcriptome, and virome sequences were annotated for taxonomic inference and gene function by using FIGfam (isofunctional homolog sequences) and SEED subsystems databases. When the beta diversities of microbial FIGfam annotations were compared, the control and carbadox communities were distinct 2 days after carbadox introduction. This effect was driven by carbadox-associated lower expression of FIGfams ( = 66) related to microbial respiration, carbohydrate utilization, and RNA metabolism ( < 0.1), suggesting bacteriostatic or bactericidal effects within certain populations. Interestingly, carbadox treatment caused greater expression of FIGfams related to all stages of the phage lytic cycle 2 days following the introduction of carbadox ( ≤0.07), suggesting the carbadox-mediated induction of prophages and phage DNA recombination. These effects were diminished by 7 days of continuous carbadox in the feed, suggesting an acute impact. Additionally, the viromes included a few genes that encoded resistance to tetracycline, aminoglycoside, and beta-lactam antibiotics but these did not change in frequency over time or with treatment. The results show decreased bacterial growth and metabolism, prophage induction, and potential transduction of bacterial fitness genes in swine gut bacterial communities as a result of carbadox administration. FDA regulations on agricultural antibiotic use have focused on antibiotics that are important for human medicine. Carbadox is an antibiotic not used in humans but frequently used on U.S. pig farms. It is important to study possible side effects of carbadox use because it has been shown to promote bacterial evolution, which could indirectly impact antibiotic resistance in bacteria of clinical importance. Interestingly, the present study shows greater prophage gene expression in feces from carbadox-fed animals than in feces from nonmedicated animals 2 days after the initiation of in-feed carbadox treatment. Importantly, the phage genetic material isolated in this study contained genes that could provide resistance to antibiotics that are important in human medicine, indicating that human-relevant antibiotic resistance genes are mobile between bacteria via phages. This study highlights the collateral effects of antibiotics and demonstrates the need to consider diverse antibiotic effects whenever antibiotics are being used or new regulations are considered.
Topics: Animal Feed; Animals; Anti-Infective Agents; Bacteriophages; Carbadox; Drug Resistance, Microbial; Feces; Gastrointestinal Microbiome; Gene Expression Profiling; Metagenome; Prophages; Sus scrofa; Swine; Transcription, Genetic
PubMed: 28790203
DOI: 10.1128/mBio.00709-17 -
Antibiotics (Basel, Switzerland) Jul 2020Antibiotics and pharmacological zinc supplementation were commonly used as growth promoters for several decades in the swine industry before being limited because of...
Antibiotics and pharmacological zinc supplementation were commonly used as growth promoters for several decades in the swine industry before being limited because of public health and environmental concerns. Further, the physiological and metabolic responses associated with their growth promotion effects are unclear. To characterize these responses induced by pharmacological zinc supplementation (2500 mg/kg) and carbadox (55 mg/kg), 192 post-weaning pigs were fed basal and test diets for 43 days. Compared with basal, pharmacological zinc and carbadox independently improved growth performance. Pharmacological zinc increased gastric mucosa thickness compared with basal zinc, while carbadox increased intestinal villus:crypt ratio compared with non-carbadox. Pharmacological zinc and carbadox independently reduced interleukin (IL)-1β concentration compared with basal zinc and non-carbadox. Pharmacological zinc increased IL-1RA:IL-1 ratio by 42% compared with basal zinc, while carbadox tended to increase the IL-10 and IL10:IL-12 ratio compared with non-carbadox. Carbadox increased fecal concentrations of histidine and lysine compared with non-carbadox. The independent effect of pharmacological zinc and carbadox on morphology and nutrient metabolism, and their shared effect on immunity may contribute to the additive effect on growth promotion. These results further confirmed the concept that growth promotion is multifactorial intervention. Therefore, elucidating growth-promoting effects and searching for alternatives should include wide-spectrum evaluation.
PubMed: 32751572
DOI: 10.3390/antibiotics9080463 -
Frontiers in Veterinary Science 2021This study investigated the use of carbadox in the diet of nursery pigs. Ten pens of weanling piglets were assigned to 2 treatments: one containing carbadox and another...
This study investigated the use of carbadox in the diet of nursery pigs. Ten pens of weanling piglets were assigned to 2 treatments: one containing carbadox and another without it. From days 21 to 35 of age, the first group of piglets was fed carbadox at 55 mg/kg of diet; followed by 27.5 mg/kg from days 36 to 49; and 0 mg/kg from days 50 to 63. The second group of pigs was fed a control diet without carbadox from days 21 to 63 of age. On days 35, 49, and 63, fecal samples were collected directly from the rectum of 2 piglets in each pen, and the samples were subjected to microbial DNA sequencing and metagenomic functional analysis using the 16S rRNA gene. Feed conversion from days 21 to 63 was improved ( = 0.04) in the group of piglets fed carbadox. Faith's phylogenetic diversity was similar ( = 0.89) for both groups of piglets on day 35, but it was diminished ( = 0.01) in the carbadox-fed group on day 49; however, following the complete removal of carbadox from their diets, this microbial diversity index was once again found to be similar ( = 0.27) in both groups on day 63. Likewise, abundances of , and were all similar between the two groups ( ≥ 0.40) on day 35, but were smaller in the carbadox group ( ≤ 0.05) on day 49; however, on day 63, abundances of all these genera were once again similar ( ≥ 0.29). Metabolic pathways involved in cellular growth, death, and genetic information processing (translation) were found to be similarly expressed in the microbiota of piglets from both groups on day 35 ( ≥ 0.52), but decreased in the carbadox group on day 49 ( ≤ 0.05), and were similar again in both groups on day 63 ( ≥ 0.51). These results revealed that feeding carbadox to piglets during the first 4 weeks after weaning significantly affected their fecal microbiotas; however, 2 weeks after the removal of carbadox, those changes tended to disappear, indicating that the shifts were carbadox-dependent.
PubMed: 33778032
DOI: 10.3389/fvets.2021.601394 -
Journal of Analytical Methods in... 2018This paper presents a convenient and sensitive LC-MS/MS method for the simultaneous determination of carbadox and olaquindox residues, including desoxyolaquindox (DOLQ),...
This paper presents a convenient and sensitive LC-MS/MS method for the simultaneous determination of carbadox and olaquindox residues, including desoxyolaquindox (DOLQ), desoxycarbadox (DCBX), quinoxaline-2-carboxylic acid (QCA), 3-methyl-quinoxaline-2-carboxylic acid (MQCA), and the glycine conjugates of QCA and MQCA (namely, QCA-glycine and MQCA-glycine, resp.) in swine muscle and liver tissues. Tissue samples were extracted with 2% metaphosphoric acid in 20% methanol and cleaned up by solid-phase extraction (SPE) on a mixed-mode anion-exchange column (Oasis MAX). Analysis was performed on a C column by detection with mass spectrometry in the multiple reaction monitoring (MRM) mode. The limits of detection (LODs) of the six analytes were determined to be 0.01 g·kg to 0.25 g·kg, and the limits of quantification (LOQs) were 0.02 g·kg to 0.5 g·kg. The total recoveries of the six analytes in all tissues were higher than 79.1% with the RSD% less than 9.2%. The developed method can determine the real residue level of QCA and MQCA, whether they are present in free form or as glycine conjugates in tissues, together with the carcinogenic desoxy metabolites DCBX and DOLQ with high recovery. Therefore, this method was suitable for routine analysis of residue control programmes and the residue depletion study of CBX and OLQ on swine.
PubMed: 30079259
DOI: 10.1155/2018/2834049 -
Translational Animal Science Apr 2021A total of 360 weanling pigs (DNA 200 × 400; initially 9.7 ± 0.23 kg BW) were used in a 21-d experiment with 6 pigs/pen, 10 replicate pens/treatment, and 2 separate...
A total of 360 weanling pigs (DNA 200 × 400; initially 9.7 ± 0.23 kg BW) were used in a 21-d experiment with 6 pigs/pen, 10 replicate pens/treatment, and 2 separate nursery rooms, each with 30 pens. Pigs were weighed and allotted to pens based on BW in a completely randomized block design to one of six treatment diets: 1) Negative control (no organic acids or antibiotics) and the control with 2) 0.25% acidifier A; 3) 0.3% acidifier B; 4) 0.5% acidifier C); 5) 50 g/ton carbadox; and 6) 400 g/ton chlortetracycline (). Upon weaning, a common diet with no antibiotics or additives was fed for 21 d (Phases 1 and 2; days -21 to 0), followed by a 21-d experimental period (Phase 3; days 0 to 21) where treatment diets were fed. Pigs and feeders were individually weighed on a weekly basis to calculate average daily gain (), average daily feed intake (), and feed efficiency (). Data were analyzed using the PROC GLIMMIX procedure of SAS (v 9.4, SAS Inst., Cary, NC) with pen as the experimental unit, treatment as a fixed effect, and room as a random effect. Dietary treatment had a significant impact ( < 0.05) on ADG, ADFI, and G:F each week and for the overall experimental period (days 0 to 21). Specifically, from days 0 to 7, pigs fed CTC had increased ( = 0.001) ADG compared with those fed acidifier B, acidifier C, and carbadox, whereas pigs fed the negative control and acidifier A diets were intermediate. Additionally, pigs fed the CTC diet had improved ( = 0.0002) ADFI when compared with all other treatments. From days 7 to 14 and days 14 to 21, pigs fed the carbadox diet had decreased ( < 0.0001) ADG compared with all other treatments. During the overall period (days 0 to 21), pigs fed diets containing carbadox had reduced ADG and ADFI ( < 0.0001), whereas pigs fed CTC had improved ( < 0.0001) ADG compared with all other treatments. Additionally, blood parameters, fecal consistency, and fecal microbial populations were analyzed on a subset of pigs ( = 5 pigs/treatment). Dietary treatment significantly affected ( < 0.05) concentrations of protein, globulin, phosphorus, alkaline phosphatase, and sorbitol dehydrogenase in the blood. Treatment also significantly impacted ( = 0.0005) fecal score but did not affect ( = 0.59) fecal microbial growth from days 0 to 21. In summary, CTC continues to be a valuable additive to improve performance in the nursery. Further investigation surrounding the efficacy of dietary acidifiers as antibiotic alternatives is warranted given inconclusive evidence in this study.
PubMed: 34085026
DOI: 10.1093/tas/txab040 -
Journal of Animal Science and... May 2022Our previous study has shown that supplementation of trace amounts of antibiotic exacerbated the detrimental effects of enterotoxigenic E. coli (ETEC) infection and...
BACKGROUND
Our previous study has shown that supplementation of trace amounts of antibiotic exacerbated the detrimental effects of enterotoxigenic E. coli (ETEC) infection and delayed the recovery of pigs that may be associated with modified metabolites and metabolic pathways. Therefore, the objective of this study was to explore the impacts of trace levels of antibiotic (carbadox) on host metabolic profiles and colon microbiota of weaned pigs experimentally infected with ETEC F18.
RESULTS
The multivariate analysis highlighted a distinct metabolomic profile of serum and colon digesta between trace amounts of antibiotic (TRA; 0.5 mg/kg carbadox) and label-recommended dose antibiotic (REC; 50 mg/kg carbadox) on d 5 post-inoculation (PI). The relative abundance of metabolomic markers of amino acids, carbohydrates, and purine metabolism were significantly differentiated between the TRA and REC groups (q < 0.2). In addition, pigs in REC group had the highest (P < 0.05) relative abundance of Lactobacillaceae and tended to have increased (P < 0.10) relative abundance of Lachnospiraceae in the colon digesta on d 5 PI. On d 11 PI, pigs in REC had greater (P < 0.05) relative abundance of Clostridiaceae compared with other groups, whereas had reduced (P < 0.05) relative abundance of Prevotellaceae than pigs in control group.
CONCLUSIONS
Trace amounts of antibiotic resulted in differential metabolites and metabolic pathways that may be associated with its slow responses against ETEC F18 infection. The altered gut microbiota profiles by label-recommended dose antibiotic may contribute to the promotion of disease resistance in weaned pigs.
PubMed: 35527278
DOI: 10.1186/s40104-022-00703-5 -
Talanta Nov 2015Carbadox (CBX) and olaquindox (OLA) were used in poultry and swine feed for growth promotion, to improve feed efficiency and increase the rate of weight gain. However,...
Carbadox (CBX) and olaquindox (OLA) were used in poultry and swine feed for growth promotion, to improve feed efficiency and increase the rate of weight gain. However, the use of these agents in feedingstuffs was prohibited because of concerns about their toxicity. Regulatory laboratories are required to have suitably validated analytical methods to ensure compliance with the ban. A quantitative and confirmatory method for determining the presence of CBX and OLA in poultry and swine feed by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed, optimized, and validated. The analytes extraction was performed with a mixture of water and acetonitrile (1:1v/v) and cleanup with hexane and C18 (dispersive phase). The method was evaluated by the following parameters: specificity, linearity, matrix effect, decision limits (CCα), detection capability (CCβ), accuracy, precision, limits of detection (LoD), limits of quantification (LoQ) and measurement uncertainty. The validated method presented a broad linear study range and no significant matrix effect. The limit of detection (LoD) was defined at 9 μg kg(-1) for CBX and 80 μg kg(-1) for OLA, and the limit of quantification (LoQ) was defined at 12 μg kg(-1) and 110 μg kg(-1) for CBX and OLA, respectively. The accuracy of the method was adequate for CBX and OLA. The recovery values found in the repeatability conditions were 99.41% for CBX and 104.62% for OLA. Under intralaboratory reproducibility conditions, the values were 98.63% for CBX and 95.07% for OLA. It was concluded that the performance parameters demonstrated total method adequacy for the detection and quantification of CBX and OLA in poultry and swine feedingstuffs.
Topics: Animal Feed; Animals; Carbadox; Chromatography, High Pressure Liquid; Food Analysis; Food Contamination; Poultry; Quinoxalines; Swine; Tandem Mass Spectrometry
PubMed: 26452885
DOI: 10.1016/j.talanta.2015.07.029 -
Molecules (Basel, Switzerland) Aug 2019A hybrid nanocomposite consisting of hydroxylated multi-walled carbon nanotubes (MWCNTs-OH) and cube mesoporous carbon (CMK-8) was applied in this study to construct an...
Novel Electrochemical Sensor Fabricated for Individual and Simultaneous Ultrasensitive Determination of Olaquindox and Carbadox Based on MWCNT-OH/CMK-8 Hybrid Nanocomposite Film.
A hybrid nanocomposite consisting of hydroxylated multi-walled carbon nanotubes (MWCNTs-OH) and cube mesoporous carbon (CMK-8) was applied in this study to construct an MWCNT-OH/CMK-8/gold electrode (GE) electrochemical sensor and simultaneously perform the electro-reduction of olaquindox (OLA) and carbadox (CBX). The respective peak currents of CBX and OLA on the modified electrode increased by 720- and 595-fold relative to the peak current of GE. The performances of the modified electrode were investigated with electrochemical impedance spectroscopy, cyclic voltammetry, and differential pulse voltammetry. Then, the modified electrodes were used for the individual and simultaneous determination of OLA and CBX. The fabricated sensor demonstrated a linear response at 0.2-500 nmol/L in optimum experimental conditions, and the detection limits were 104.1 and 62.9 pmol/L for the simultaneous determination of OLA and CBX, respectively. As for individual determination, wide linear relationships were obtained for the detected OLA with levels of 0.05-500 nmol/L with LOD of 20.7 pmol/L and the detected CBX with levels of 0.10-500 nmol/L with LOD of 50.2 pmol/L. The fabricated sensor was successfully used in the independent and simultaneous determination of OLA and CBX in spiked pork samples.
Topics: Carbadox; Electrochemical Techniques; Electrodes; Gold; Membranes, Artificial; Nanocomposites; Nanotubes, Carbon; Quinoxalines; Spectrum Analysis
PubMed: 31443345
DOI: 10.3390/molecules24173041 -
Metabolites Jan 2021This study explored the metabolomic profiles in ileal mucosa and colon digesta in response to enterotoxigenic F18 (ETEC) infection and dietary use of probiotics and...
This study explored the metabolomic profiles in ileal mucosa and colon digesta in response to enterotoxigenic F18 (ETEC) infection and dietary use of probiotics and low-dose antibiotics. Weaned pigs ( = 48, 6.17 ± 0.36 kg body weight) were randomly allotted to one of four treatments. Pigs in the negative control (NC) were fed a basal diet without ETEC challenge, whereas pigs in the positive control (PC), antibiotic, and probiotic groups were fed the basal diet, basal diet supplemented with 50 mg/kg of carbadox, or 500 mg/kg of , respectively, and orally challenged with ETEC F18. All pigs were euthanized at day 21 post-inoculation to collect ileal mucosa and colon digesta for untargeted metabolomic profiling using gas chromatography coupled with time-of-flight mass spectrometry. Multivariate analysis highlighted a more distinct metabolomic profile of ileal mucosa metabolites in NC compared to the ETEC-challenged groups. The relative abundance of 19 metabolites from the ileal mucosa including polyamine, nucleotide, monosaccharides, fatty acids, and organic acids was significantly different between the NC and PC groups ( < 0.1). In colon digesta, differential metabolites including 2-monoolein, lactic acid, and maltose were reduced in the carbadox group compared with the probiotics group. In conclusion, several differential metabolites and metabolic pathways were identified in ileal mucosa, which may suggest an ongoing intestinal mucosal repair in the ileum of ETEC-challenged pigs on day 21 post-inoculation.
PubMed: 33573321
DOI: 10.3390/metabo11020081