Swine growth promotion with antibiotics or alternatives can increase antibiotic resistance gene mobility potential.

Even though the use of antibiotics for food-producing animals may contribute to the emergence of antimicrobial resistance, antibiotics are still used as growth promoters. Due to consumer and regulatory pressures, the use of alternatives to antibiotics as growth promoters is increasing, thus more information is needed on their capability to disseminate antimicrobial resistance compared to antibiotics. We investigated the impacts of carbadox (antibiotic), copper sulfate and zinc oxide (metals) and mushroom powder (natural product) on the pig fecal resistome and microbiome. Antibiotic resistance gene (ARG) and mobile genetic element (MGE) abundances were measured using a high-throughput qPCR array with 382 primer pairs. Bacterial community composition was determined by 16S rRNA gene sequencing. More ARGs co-occurred with MGEs in the growth promoter group samples than in the control group samples. Community composition could not be linked to resistome in the growth promoter group samples, indicating a potential decoupling of ARGs and phylogeny. Additionally, machine-learning methods aided in defining the community and resistome differences in response to treatments. Since increased ARG mobility potential was the primary response to the dietary additives used in this study, we suggest that ARG mobility should be considered when designing antimicrobial use policies and antimicrobial resistance surveillances.

Bacillus subtilis: a potential growth promoter in weaned pigs in comparison to carbadox.

The study was conducted to investigate the efficacy of a probiotic Bacillus subtilis strain on growth performance, diarrhea, systemic immunity, and intestinal health of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli and to compare the efficacy of B. subtilis with that of carbadox. Weaned pigs (n = 48, 6.17 ± 0.36 kg body weight [BW]) were individually housed in disease containment rooms and randomly allotted to one of four dietary treatments: negative control (NC, control diet without E. coli challenge), positive control (PC, control diet with E. coli challenge), and supplementation of 50 mg/kg of carbadox (antibiotic growth promotor [AGP]) or 2.56 × 109 CFU/kg of B. subtilis probiotics (PRO). The experiment lasted for 28 d with 7 d before and 21 d after the first E. coli inoculation. Fecal and blood samples were collected on days 0, 3, 7, 14, and 21 post inoculation (PI) to analyze ß-hemolytic coliforms and complete blood cell count, respectively. Diarrhea score was recorded daily for each pig to calculate the frequency of diarrhea. All pigs were euthanized at day 21 PI to collect jejunal and ileal mucosa for gene expression analysis. Pigs in AGP had greater (P < 0.05) BW on days 7, 14, and 21 PI than pigs in PC and PRO groups. Supplementation of PRO enhanced pigs' BW on day 21 PI compared with the PC. Escherichia coli F18 challenge reduced (P < 0.05) average daily gain (ADG) and feed efficiency from day 0 to 21 PI, while supplementation of carbadox or PRO enhanced ADG and feed efficiency in E. coli F18-challenged pigs from day 0 to 21 PI. Pigs in AGP and PRO groups had reduced (P < 0.05) frequency of diarrhea throughout the experiment and fecal ß-hemolytic coliforms on day 7 PI than pigs in the PC. Pigs in PRO had greater (P < 0.05) gene expression of CLDN1 in jejunal mucosa than pigs in the PC. Supplementation of carbadox or PRO reduced (P < 0.05) the gene expression of IL6 and PTGS2 in ileal mucosa of E. coli-infected pigs compared with pigs in the PC. Pigs in the PRO group had lower (P < 0.05) white blood cell number and neutrophil count, and serum haptoglobin concentration on day 7 PI, and less (P < 0.05) monocyte count on day 14 PI, compared with PC. In conclusion, supplementation of probiotic B. subtilis could enhance disease resistance and promote the growth performance of weaned pigs under disease challenge conditions. The potential mechanisms include but not limited to enhanced gut barrier integrity and local and systemic immune responses of weaned pigs.

Discovery of novel antigiardiasis drug candidates.

Giardiasis is a severe intestinal parasitic disease caused by Giardia lamblia, which inflicts many people in poor regions and is the most common parasitic infection in the United States. Current standard care drugs are associated with undesirable side effects, treatment failures, and an increasing incidence of drug resistance. As follow-up to a high-throughput screening of an approved drug library, which identified compounds lethal to G. lamblia trophozoites, we have determined the minimum lethal concentrations of 28 drugs and advanced 10 of them to in vivo studies in mice. The results were compared to treatment with the standard care drug, metronidazole, in order to identify drugs with equal or better anti-Giardia activities. Three drugs, fumagillin, carbadox, and tioxidazole, were identified. These compounds were also potent against metronidazole-resistant human G. lamblia isolates (assemblages A and B), as determined in in vitro assays. Of these three compounds, fumagillin is currently an orphan drug used within the European Union to treat microsporidiosis in immunocompromised individuals, whereas carbadox and tioxidazole are used in veterinary medicine. A dose-dependent study of fumagillin in a giardiasis mouse model revealed that the effective dose of fumagillin was ∼ 100-fold lower than the metronidazole dose. Therefore, fumagillin may be advanced to further studies as an alternative treatment for giardiasis when metronidazole fails.

Controlling Salmonella infection in weanling pigs through water delivery of direct-fed microbials or organic acids: Part II. Effects on intestinal histology and active nutrient transport.

The objective of this study was to evaluate the effects of water-delivered, direct-fed microbials (DFM) or organic acids on intestinal morphology and active nutrient absorption in weanling pigs after deliberate Salmonella infection. Pigs (n = 88) were weaned at 19 ± 2 d of age and assigned to 1 of the following treatments, which were administered for 14 d: 1) control diet; 2) control diet + DFM (Enterococcus faecium, Bacillus subtilis, and Bacillus licheniformis) in drinking water at 10(9) cfu/L for each strain of bacteria; 3) control diet + organic acid-based blend (predominantly propionic, acetic, and benzoic acids) in drinking water at 2.58 mL/L; and 4) control diet + 55 mg/kg carbadox. Pigs were challenged with 10(10) cfu Salmonella enterica var Typhimurium 6 d after commencement of treatments. Pigs (n = 22/d) were harvested before Salmonella challenge and on d 2, 4, and 8 after challenge. Duodenal, jejunal, and ileal mucosal tissues were sampled for measurement of villus height and crypt depth. Jejunal tissue was sampled for determination of active nutrient absorption in modified Ussing chambers. Duodenal villus height was greater in pigs fed in-feed antibiotic before infection (P < 0.05). Jejunal crypts were deeper in DFM- and acid-treated pigs on d 4 after infection compared with all other treatments (P < 0.05). Salmonella infection resulted in a linear decrease in phosphorus (P < 0.001) and glucose (P < 0.05) active transport, and an increase (P < 0.001) in glutamine uptake immediately after challenge. Salmonella infection reduced basal short-circuit current (I(sc)); however, water-delivered DFM or organic acid treatments caused greater basal I(sc) on d 2 after challenge than did carbadox. Carbachol-induced chloride ion secretion was greatest in negative control pigs before infection (P < 0.01) and DFM-treated pigs (P < 0.05) after infection. In conclusion, both the DFM and acidification treatments induced increases in basal active ion movement and jejunal crypt depth, which could be interpreted as responses consistent with increased Salmonella pathology, but none of the additives markedly affected intestinal absorptive and secretory function in response to Salmonella challenge.

Controlling Salmonella infection in weanling pigs through water delivery of direct-fed microbials or organic acids. Part I: effects on growth performance, microbial populations, and immune status.

Pigs (n = 88) weaned at 19 ± 2 d of age were used in a 14-d study to evaluate the effects of water-delivered direct-fed microbials (DFM) or organic acids on growth, immune status, Salmonella infection and shedding, and intestinal microbial populations after intranasal inoculation of Salmonella Typhimurium (10(10) cfu/pig). Pigs were challenged with Salmonella 6 d after commencement of water treatments. Treatments were 1) control diet; 2) control diet + DFM (Enterococcus faecium, Bacillus subtilis, and Bacillus licheniformis) in drinking water at 10(9) cfu/L for each strain of bacteria; 3) control diet + an organic acid-based blend (predominantly propionic, acetic, and benzoic acid) in drinking water at 2.58 mL/L; and 4) control diet + 55 mg/kg of carbadox. Serum samples were taken on d 6, 8, 10, and 14 for determination of tumor necrosis factor α (TNFα) concentrations. Fecal samples were taken on d 0, 5, 7, and 11 for determination of Salmonella shedding and enumeration of coliforms. Pigs were euthanized on d 6, 8, 10, and 14. Intestinal and cecal tissue and digesta and mesenteric lymph nodes were sampled and analyzed for Salmonella. Duodenal, jejunal, and ileal mucosal scrapings were sampled for measurement of mucosal TNFα concentrations. Water delivery of DFM prevented a decline in ADG on d 2 to 6 postchallenge compared with the negative control (P < 0.05). Coliform counts tended to be greater (P = 0.09) in the cecum of the DFM treatment group on d 2 postinfection compared with the negative control and acid treatment groups. However, Salmonella prevalence in the feces, gastrointestinal tract, or lymph nodes was not affected by water delivery of acids or DFM. Serum and mucosal TNFα concentrations were not affected by treatment throughout the study with the exception of ileal concentrations on d 4 postchallenge, which were greater in the negative control group compared with all other treatments (P < 0.05). The in-feed antibiotic was the only treatment that reduced Salmonella prevalence and this was localized to the cecum on d 8 postinfection. In conclusion, the DFM and organic acid treatments used in this study offered little or no benefits to pigs infected with Salmonella and should not be considered under the constraints of this study as viable alternatives to in-feed antibiotics in a pathogen challenge situation.

Antibiotics in feed induce prophages in swine fecal microbiomes.

UNLABELLED: Antibiotics are a cost-effective tool for improving feed efficiency and preventing disease in agricultural animals, but the full scope of their collateral effects is not understood. Antibiotics have been shown to mediate gene transfer by inducing prophages in certain bacterial strains; therefore, one collateral effect could be prophage induction in the gut microbiome at large. Here we used metagenomics to evaluate the effect of two antibiotics in feed (carbadox and ASP250 [chlortetracycline, sulfamethazine, and penicillin]) on swine intestinal phage metagenomes (viromes). We also monitored the bacterial communities using 16S rRNA gene sequencing. ASP250, but not carbadox, caused significant population shifts in both the phage and bacterial communities. Antibiotic resistance genes, such as multidrug resistance efflux pumps, were identified in the viromes, but in-feed antibiotics caused no significant changes in their abundance. The abundance of phage integrase-encoding genes was significantly increased in the viromes of medicated swine over that in the viromes of nonmedicated swine, demonstrating the induction of prophages with antibiotic treatment. Phage-bacterium population dynamics were also examined. We observed a decrease in the relative abundance of Streptococcus bacteria (prey) when Streptococcus phages (predators) were abundant, supporting the "kill-the-winner" ecological model of population dynamics in the swine fecal microbiome. The data show that gut ecosystem dynamics are influenced by phages and that prophage induction is a collateral effect of in-feed antibiotics. IMPORTANCE: This study advances our knowledge of the collateral effects of in-feed antibiotics at a time in which the widespread use of "growth-promoting" antibiotics in agriculture is under scrutiny. Using comparative metagenomics, we show that prophages are induced by in-feed antibiotics in swine fecal microbiomes and that antibiotic resistance genes were detected in most viromes. This suggests that in-feed antibiotics are contributing to phage-mediated gene transfer, potentially of antibiotic resistance genes, in the swine gut. Additionally, the so-called "kill-the-winner" model of phage-bacterium population dynamics has been shown in aquatic ecosystems but met with conflicting evidence in gut ecosystems. The data support the idea that swine fecal Streptococcus bacteria and their phages follow the kill-the-winner model. Understanding the role of phages in gut microbial ecology is an essential component of the antibiotic resistance problem and of developing potential mitigation strategies.

Growth performance and gastrointestinal microbial ecology responses of piglets receiving Saccharomyces cerevisiae fermentation products after an oral challenge with Escherichia coli (K88).

The effects of Saccharomyces cerevisiae fermentation products (YFP) on growth performance and gastrointestinal (GIT) microbial ecology in 90 weanling pigs orally challenged with Escherichia coli K88(+) (ETEC) were investigated. The YFP were an original YFP product (XPC) and a water-suspendable yeast fermentation prototype (WSYFP) from a commercial company. Treatments consisted of a negative control (NC, no in-feed or in-water additive), carbadox (AB, 55 mg of carbadox/kg of feed), XPC (in feed, 0.2%), and WSYFP (in water, 0.5, 1, or 2 g/pig per day), and each was allotted to 5 pens (3 pigs/pen). The diets met the 1998 NRC specifications. Pigs were acclimated to treatments for a 7-d period before an ETEC challenge. On d 8, blood was collected from pigs to determine the baseline packed cell volume (PCV) measurement, and pigs were orally challenged with ETEC. At various time points postchallenge, blood samples were taken, performance measures and fecal consistency scores were recorded, and gut digesta and tissue samples were taken to evaluate GIT morphology, microbial ecology, and metabolites. Preplanned contrasts were used for comparison. Pigs receiving YFP had greater ADFI than NC pigs on d 3 (424 vs. 378 g/d; P = 0.01) and d 7 (506 vs. 458 g/d; P = 0.03) postchallenge. This effect of YFP on ADFI was similar to that of AB on d 3, but pigs receiving AB ate more (576 vs. 506 g/d; P = 0.03) at d 7 than pigs receiving YFP. Pigs exhibited reduced (P < 0.001) PCV upon ETEC challenge; however, pigs receiving additives sustained a greater (P < 0.05) PCV at 72 h compared with the NC group. Compared with the NC pigs, pigs receiving YFP showed a smaller (P < 0.05) number of ileal mucosa adherent ETEC and prevalence of the order Enterobacteriales in the ileal digesta, which corresponded to less (5.09 vs. 6.97 mg/dL; P = 0.03) colonic ammonia on d 7 postchallenge. Most of the indices for ileal digesta bacterial richness and diversity were greater (P < 0.01) for YFP pigs compared with NC pigs. However, results also indicated that the influence of YFP on the piglet intestinal microenvironment might differ when given in feed or water during ETEC challenge. In conclusion, pigs receiving YFP showed a better appetite in the presence of ETEC, which, together with the greater ileal digesta bacteria richness and diversity and decreased ETEC adhering to the mucosa and reduced colonic ammonia, indicates a healthier GIT environment.

The effects of dietary additives on faecal levels of Lactobacillus spp., coliforms, and Escherichia coli, and faecal prevalence of Salmonella spp. and Campylobacter spp. in US production nursery swine.

AIMS: In the United States, carbadox and copper sulfate are growth promoters commonly used in combination in nursery swine diets. Our aim was to determine how selected dietary additives affect selected bacterial populations and pathogens in nursery swine, and compare to larch extract, which contains potential antibacterial activities. METHODS AND RESULTS: Piglets were weaned and sorted into one of the four treatments: (i) basal diet without antimicrobials; (ii) basal diet with carbadox + copper sulfate; (iii) basal diet + 1000 ppm larch extract; or (iv) basal diet + 2000 ppm larch extract. Diets were fed for a 4-week period after weaning. In both trials, the carbadox + copper sulfate group consumed more feed over the 4-week period relative to the other three diet groups (P < 0.05), but did not gain significantly more weight. Faecal shedding of Salmonella spp. was not affected by dietary supplement in either trial, but faecal shedding of Campylobacter spp. was the lowest for the carbadox + copper sulfate diet. In faecal samples collected at the end of each trial, Lactobacillus spp. cell counts for the basal and larch extract diets were nearly 1.0 log(10) g(-1) faeces greater (P < 0.05) than the carbadox + copper sulfate group, whereas the coliforms and Escherichia coli were nearly 1.0 log(10) g(-1) faeces lower (P < 0.05). CONCLUSIONS: Compared to basal fed animals, supplementation with carbadox + copper sulfate significantly altered faecal E. coli, coliform bacteria and Lactobacillus spp. Larch extract has no benefit up to 0.2% of diet in regard to pathogen shedding, whereas carbadox + copper sulfate decreased faecal shedding of Campylobacter spp. SIGNIFICANCE AND IMPACT OF THE STUDY: Current swine management practices in the United States may be beneficial to managing Campylobacter spp. shedding in nursery swine, but also result in significant changes in the resident gastrointestinal microflora.

Effect of supplementing zinc oxide and biotin with or without carbadox on nursery pig performance.

A 28-d nursery experiment was conducted to evaluate the effects of supplementing zinc oxide and biotin with or without a feed-grade antimicrobial agent (carbadox) on nursery pig performance, and plasma and fecal Zn concentrations. One hundred ninety-two crossbred pigs (initial BW = 5.94 +/- 0.03 kg; age = 17 +/- 2 d) were weaned and allotted to 1 of 8 dietary treatments based on BW, sex, and ancestry in a randomized complete block design (3 pigs/pen and 8 replications). Dietary treatments consisted of supplementation of ZnO at 0 or 3,000 mg/kg, d-biotin at 0 or 440 microg/kg, and carbadox at 0 or 55 mg/kg of diets in a 2 x 2 x 2 factorial arrangement of treatments. Phase 1 (d 0 to 14) and phase 2 (d 14 to 28) nursery diets were fed in meal form. Fecal samples were collected weekly, and blood samples were collected at d 0, 14, and 28 to determine fecal and plasma Zn concentrations, respectively. The basal diet contained 165 mg/kg of Zn as ZnSO(4) and 220 microg/kg biotin as d-biotin. Pigs supplemented with 440 microg/kg of d-biotin, independent of antibiotic and ZnO additions, had greater overall ADG (P = 0.02) than pigs fed no supplemental d-biotin postweaning. Overall ADG, ADFI, and G:F were not affected when pigs were supplemented with 3,000 mg/kg of Zn as ZnO or 55 mg/kg of carbadox. When pigs were fed 55 mg/kg of carbadox without supplemental biotin, plasma Zn concentration was less, whereas when biotin and carbadox were supplemented to nursery pig diets, plasma Zn concentrations did not decrease as with feeding carbadox alone (biotin x carbadox, P < 0.001). During wk 2, pigs fed 3,000 mg/kg of Zn as ZnO and 440 microg/kg of d-biotin had greater fecal Zn concentrations than pigs fed diets with only 3,000 mg/kg of Zn as ZnO (Zn x biotin, P = 0.04). In addition, pigs supplemented with 3,000 mg/kg of Zn as ZnO in combination with carbadox and d-biotin had greater fecal Zn concentrations compared with pigs fed diets containing no additional Zn during wk 2 (Zn x biotin x carbadox, P = 0.04). Fecal Zn concentrations increased over time (P < 0.001), and pigs fed supplemental ZnO had greater (P < 0.001) fecal Zn concentrations. These results indicate that feeding nursery pigs 440 microg/kg of d-biotin improved ADG, independent of zinc and carbadox supplementation, as well as increasing Zn excretion.

Collateral effects of antibiotics: carbadox and metronidazole induce VSH-1 and facilitate gene transfer among Brachyspira hyodysenteriae strains.

Brachyspira hyodysenteriae is an anaerobic spirochete and the etiologic agent of swine dysentery. The genome of this spirochete contains a mitomycin C-inducible, prophage-like gene transfer agent designated VSH-1. VSH-1 particles package random 7.5-kb fragments of the B. hyodysenteriae genome and transfer genes between B. hyodysenteriae cells. The chemicals and conditions inducing VSH-1 production are largely unknown. Antibiotics used in swine management and stressors inducing traditional prophages might induce VSH-1 and thereby stimulate lateral gene transfer between B. hyodysenteriae cells. In these studies, VSH-1 induction was initially detected by a quantitative real-time reverse transcriptase PCR assay evaluating increased transcription of hvp38 (VSH-1 head protein gene). VSH-1 induction was confirmed by detecting VSH-1-associated 7.5-kb DNA and VSH-1 particles in B. hyodysenteriae cultures. Nine antibiotics (chlortetracycline, lincomycin, tylosin, tiamulin, virginiamycin, ampicillin, ceftriaxone, vancomycin, and florfenicol) at concentrations affecting B. hyodysenteriae growth did not induce VSH-1 production. By contrast, VSH-1 was detected in B. hyodysenteriae cultures treated with mitomycin C (10 microg/ml), carbadox (0.5 microg/ml), metronidazole (0.5 microg/ml), and H(2)O(2) (300 microM). Carbadox- and metronidazole-induced VSH-1 particles transmitted tylosin and chloramphenicol resistance determinants between B. hyodysenteriae strains. The results of these studies suggest that certain antibiotics may induce the production of prophage or prophage-like elements by intestinal bacteria and thereby impact intestinal microbial ecology.

Effect of sodium butyrate on growth performance and response to lipopolysaccharide in weanling pigs.

Two experiments were conducted to determine the effects of dietary sodium butyrate on growth performance and response to Escherichia coli lipopolysaccharide (LPS) in weanling pigs. In a 28-d experiment, 180 pigs (initial BW 6.3 kg) were fed 0, 0.05, 0.1, 0.2, or 0.4% sodium butyrate, or 110 mg/kg of dietary tylosin. There was no effect of dietary sodium butyrate or tylosin on overall G:F, but there was a linear trend (P < 0.07) toward decreased ADFI and ADG as levels of sodium butyrate increased. In a second 28-d experiment, 108 pigs (initial BW 6.3 kg) were assigned to 1 of 3 dietary treatments: 1) no antibiotics, 2) 0.2% sodium butyrate, or 3) 55 mg/kg of carbadox. On d 14, a subset of pigs from the no-antibiotic and butyrate treatment groups was challenged with E. coli LPS or injected with sterile saline in a 2 x 2 factorial arrangement (+/-LPS challenge; +/-dietary butyrate; n = 6 pigs/treatment group). Four hours after LPS challenge, blood samples were obtained, and samples of LM, liver, and ileum were collected for gene expression analysis. Serum samples were analyzed for IL-6, tumor necrosis factor alpha (TNFalpha), alpha(1)-acid glycoprotein, cortisol, IGF-I, insulin, and metabolites. The relative abundance of tissue cytokine and IGF-I mRNA was measured by real-time PCR. Feeding diets containing sodium butyrate or carbadox did not alter ADG or ADFI compared with pigs fed the control diet. From d 0 to 14, pigs fed diets containing 0.2% sodium butyrate had decreased (P < 0.05) ADG and tended (P < 0.06) to have decreased G:F compared with animals fed diets containing carbadox. Challenge with LPS increased (P < 0.05) serum cytokines and cortisol and decreased (P < 0.05) serum glucose and triglycerides. Injection with LPS increased (P < 0.05) the relative abundance of hepatic IL-6 and TNFalpha mRNA, increased (P < 0.05) LM TNFalpha mRNA content, and decreased (P < 0.05) IGF-I mRNA in LM. For serum cortisol, there was an interaction (P < 0.05) between dietary butyrate and LPS. The increase in serum cortisol attributable to LPS was greater (P < 0.05) in pigs fed butyrate than in pigs fed the control diet. There tended (P < 0.10) to be an interaction between LPS and diet and for butyrate to increase the relative abundance of IL-6 mRNA in LM. Carbadox did not alter cytokine or IGF-I mRNA or serum metabolites, but did decrease (P < 0.05) serum TNFalpha. These data indicate that dietary sodium butyrate does not enhance growth performance, but may regulate the response to inflammatory stimuli in weanling pigs.

Effects of water and diet acidification with and without antibiotics on weanling pig growth and microbial shedding.

Two 5-wk experiments were conducted to determine the effects of water and diet acidification with and without antibiotics on weanling pig growth performance and microbial shedding. In Exp. 1, 204 pigs (19.2 d of age) were used in a 3 x 2 factorial, with 3 dietary treatments fed with or without water acidification (2.58 mL/L of a propionic acid blend; KEM SAN, Kemin Americas, Des Moines, IA). Dietary treatments were: 1) control, 2) control + 55 ppm of carbadox (CB), and 3) dietary acid [DA; control + 0.4% organic acid-based blend (fumaric, lactate, citric, propionic, and benzoic acids; Kemin Americas)] on d 0 to 7 followed by 0.2% inorganic acid-based blend (phosphoric, fumaric, lactic, and citric acids; Kemin Americas) on d 7 to 34. In Exp. 2, 210 pigs (average 18.3 d of age) were fed 1 of 3 dietary treatments: 1) control, 2) control + 55 ppm of CB, and 3) control + 38.6 ppm of tiamulin + 441 ppm of chlortetracycline on d 0 to 7 followed by 110 ppm of chlortetracycline on d 7 to 35 (TC) with or without dietary acidification (same as Exp. 1) in a 3 x 2 factorial arrangement of treatments. For both experiments, the pigs were allotted based on genetics, sex, and initial BW [5.5 kg (Exp. 1) or 5.6 kg (Exp. 2)]. Pigs were housed at 6 or 7 (Exp. 1) and 7 (Exp. 2) pigs/pen. Treatments were fed in 3 phases: d 0 to 7, 7 to 21, and 21 to 35 (34 d, Exp. 1). Fecal grab samples were collected from 3 pigs/pen on d 6, 20, and 33 for measurement of pH and Escherichia coli. During phase 3 and overall in Exp. 1, pigs fed CB had greater (P < 0.001) ADG (overall ADG, 389 vs. 348, and 348 g/d, respectively), ADFI (P < 0.007, 608 vs. 559, and 554 g/d, respectively), and d 34 BW (P < 0.001, 18.8 vs. 17.3, and 17.3 kg, respectively) than pigs fed NC and DA. Phase 3 ADG was improved (P < 0.01) by water acidification across all diets. In Exp. 2, pigs fed CB and TC had greater ADG (P < 0.004; 315 and 303 vs. 270 g/d, respectively), ADFI (P < 0.01), and d 35 BW (P < 0.002; 16.7 and 16.2 vs. 15.1 kg, respectively) than pigs fed NC. There was a tendency (P < 0.08) for an improvement in ADG when DA was added to the NC or TC, but decreased ADG when DA was added to CB.

Effects of Acid LAC and Kem-Gest acid blends on growth performance and microbial shedding in weanling pigs.

Weanling pigs with mean initial BW of 6.04 kg (Exp.1) and 5.65 kg (Exp. 2) and mean age at weaning of 18.2 d (Exp. 1) and 17.7 d (Exp. 2) were used in two 5-wk experiments (Exp. 1, n = 180; Exp. 2, n = 300) to evaluate the effects of an organic acid blend (Acid LAC, Kemin Americas Inc., Des Moines, IA) and an inorganic/organic acid blend (Kem-Gest, Kemin Americas Inc.) on weanling pig growth performance and microbial shedding. In Exp. 1, the 5 dietary treatments were 1) negative control, 2) diet 1 + 55 ppm carbadox, 3) diet 1 + 0.4% Acid LAC, 4) diet 1 + 0.2% Kem-Gest, 5) diet 1 + 0.4% Acid LAC and 0.2% Kem-Gest. In Exp. 2, the 6 dietary treatments were diets 1 through 4 corresponding to Exp. 1, plus 5) sequence 1: 0.4% Acid LAC for 7 d followed by 0.2% Kem-Gest for 28 d, and 6) sequence 2: 0.2% Kem-Gest for 7 d followed by 0.4% Acid LAC for 28 d. Pigs were housed at 6 (Exp. 1) or 10 (Exp. 2) pigs/pen. Treatments were fed throughout the experiment in 3 phases: d 0 to 7, d 7 to 21, and d 21 to 35. In Exp. 1, there were no differences (P > 0.05) in ADG, ADFI, or G:F among the dietary treatments at any time during the study. In Exp. 2, throughout the study, pigs fed carbadox (diet 2) and sequence 1 (diet 5) diets had the greatest ADG (d 0 to 35; 262, 294, 257, 257, 292, and 261 g/d, diets 1 through 6, respectively; P < 0.05), greater ADFI than all other acid treatments (P < 0.05), and tended to have greater ADFI than diet 1 (P < 0.10). Fecal pH, Escherichia coli concentrations, and Salmonella presence were determined at d 6, 20, and 34 for Exp. 1, and on d 32 for Exp. 2. For both experiments, there was no effect of treatment on the presence of fecal Salmonella (P > 0.10) at any sampling time. In Exp. 1, fecal E. coli concentrations for pigs fed the carbadox (P < 0.05) diet were greater than for pigs fed the combination diet with 0.4% Acid LAC and 0.2% Kem-Gest on d 34, and the pigs fed the negative control diet tended (P < 0.10) to have greater fecal E. coli concentrations than those fed the combination diet on d 34. In Exp. 2, fecal pH of pigs fed sequence 1 tended to be greater than fecal pH of pigs fed diet 1, diet 4, or sequence 2 (P < 0.10), but there was no dietary effect on fecal E. coli. In Exp. 1, growth performance of pigs fed the Acid LAC and Kem-Gest diets was similar to each other and to that of the carbadox-fed pigs. Adding the combination of 0.4% Acid LAC and 0.2% Kem-Gest to nursery pig diets reduced ADFI and pig growth rate. In Exp. 2, pigs fed the acid sequence of Acid LAC-Kem-Gest had similar growth performance to pigs fed carbadox, and this novel dietary acid sequence may have merit as a replacement for antibiotics in the nursery phase.

Antibacterials that are used as growth promoters in animal husbandry can affect the release of Shiga-toxin-2-converting bacteriophages and Shiga toxin 2 from Escherichia coli strains.

Antibiotics are commonly used as growth promoters in animal husbandry worldwide. This practice has been linked to the emergence of particular antibiotic-resistant bacteria, and is now controversial. In this study, the ability of growth-promoting antibiotics to induce Shiga toxin (Stx)-converting bacteriophages from Stx-producing Escherichia coli (STEC) strains was investigated. Subinhibitory concentrations of the antibacterial growth promoters olaquindox, carbadox, tylosin and monensin were used for induction experiments. The amount of mature Stx-converting phage particles released from induced and non-induced cultures was determined, and the production of Stx was simultaneously measured by ELISA. Whereas the quinoxaline-1,4-dioxide-type antibiotics olaquindox and carbadox enhanced the release of Stx-converting phage particles from STEC cells, tylosin and monensin decreased phage induction. The production of Stx increased or decreased simultaneously with the amount of free phages. The results of this study show that particular antibacterial growth promoters can induce Stx phages. In vivo induction of Stx phages from lysogenic STEC may increase the amount of free phages in the intestine and therefore may contribute to the spread of STEC and development of new STEC pathotypes.

Incidence of antibiotic resistance in fecal Escherichia coli isolated from commercial swine farms.

Sows and pigs from 10 commercial swine farms were sampled to determine patterns of resistance of fecal Escherichia coli to five commonly used antibiotics. Before testing, farms were categorized as high or low antibiotic use based on interviews with the respective producers. On each farm, fecal swabs were obtained from five sows at 7 d postpartum and from five pigs from each sow at 7, 35, and 63 d of age. A total of 6,296 E. coli isolates from 750 pig fecal samples and a total of 462 E. coli isolates from 50 sow fecal samples were tested against apramycin, carbadox, gentamicin, neomycin, and oxytetracycline using a standardized disk diffusion test. Percentage of resistant organisms was compared between pigs at the various stages of growth, between sows and their respective pigs, and between farms of high and low antibiotic use. Incidence of resistance was greater (P < .05) in pigs at 35 or 63 d of age compared with 7 d of age for most antibiotics, and patterns differed between high-use and low-use farms. Resistance differed (P < .05) among E. coli from pigs compared to their respective sows for apramycin and carbadox on low-use farms, whereas incidence of resistance on high-use farms differed (P < .05) between sows and pigs for apramycin and oxytetracycline. These data indicate that patterns of antibiotic resistance are dependent on age of pig and level of antibiotic use.

Sensitivity of strains of Serpulina hyodysenteriae isolated in Hungary to chemotherapeutic drugs.

The sensitivity of 332 strains of Serpulina hyodysenteriae isolated in Hungary between 1978 and 1992 was tested against seven chemotherapeutic drugs frequently used for the treatment of swine dysentery, and the changes in the patterns of resistance were also monitored. All the strains remained sensitive to carbadox, with minimum inhibitory concentrations (MIC) of only 0.05 to 0.40 microgram/ml at present. The susceptibility of the strains to dimetridazole has gradually decreased, but about half of the strains are still sensitive, with large numbers of "moderately sensitive' strains; the MIC values varied within wide limits (0.1 to 50 micrograms/ml). Most of the strains were resistant to tylosin, with MIC values from 0.1 to 100 micrograms/ml. The number of strains resistant to lincomycin has gradually increased, but about half of the strains remain sensitive; the MIC values ranged from 0.2 to 100 micrograms/ml. Recently, tiamulin has proved the most effective antibiotic, but some resistant strains have already emerged (MIC values 0.05 to 50 micrograms/ml). Monensin was good for the prevention of swine dysentery, but resistance may evolve quickly; the MIC values ranged from 0.4 to 25 micrograms/ml. For sedecamycin, the MIC values (6.25 to 100 micrograms/ml) were much higher than expected.

Effects of carbadox, copper, or Yucca shidigera extract on growth performance and visceral weight of young pigs.

Two identical trials, each with 128 crossbred weanling barrows (6.8 +/- .2 and 7.6 +/- .2 kg for Trials 1 and 2, respectively), were conducted. In each trial, pigs were allowed to consume ad libitum a 16% CP corn-soybean meal basal diet (B), B + 55 ppm of carbadox, B + 250 ppm of copper (Cu), or B + 125 ppm of Yucca shidigera extract for 56 d (four pens/diet; eight pigs/pen). At d 56 of the test, eight pigs/diet (two pigs/pen) were slaughtered for carcass and viscera measurements. Data of carbadox treatment in Trial 1 are excluded from this paper because of an error in mixing of the diet. In Trial 1, during the test period from d 0 to 28, pigs fed Cu had greater (P < .05) ADG and ADFI than those fed B, whereas pigs fed yucca extract had higher ADFI (P < .05) but similar ADG (P > .05) compared with those fed B. During the period from 29 to 56 d, ADG was similar among pigs fed different diets (P > .05) but ADFI was affected (P < .05) by diet (yucca extract > Cu > B). No differences (P > .05) among diets were detected for gain/feed (G/F) and visceral weights expressed as a percentage of slaughter BW. In Trial 2, during the first 28 d, the ADG, ADFI, and G/F responses of pigs to Cu, yucca extract, and B were similar to those observed in Trial 1.(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of carbadox on fasting oxygen consumption by portal vein-drained organs and by the whole animal in growing pigs.

Fasting O2 consumption by the whole animal (W) and by portal vein-drained organs (PVDO) during the 24- to 30-h postprandial period were measured in seven growing pigs (36.1 +/- 2.3 kg) with catheters chronically placed in the hepatic portal vein, ileal vein, and carotid artery trained to consume 1.2 kg of a 16% CP corn soybean meal basal diet (B) once daily. The pigs were placed individually into an open-circuit, indirect calorimeter and connected to an arteriovenous (A-V) O2 difference analyzer for hourly simultaneous measurements of O2 consumption by W and PVDO. The PVDO O2 consumption was calculated by multiplying the A-V O2 difference by the portal vein blood flow rate derived from constant infusion of a p-aminohippuric acid solution into the ileal vein. After the initial series of hourly measurements, four pigs remained on the B diet and three pigs were fed a B + 55 ppm carbadox diet. Seven days later, the second series of measurements was made. In pigs fed the diet with carbadox added, the hourly W O2 consumptions were not different (P greater than .05) between the initial and second series and averaged 7.5 mL.min-1.kg of BW-1. However, the A-V O2 differences (mL/dL) were reduced (P less than .05) from 4.6 to 4.0 at 24 h, 4.8 to 4.0 at 25 h, and 4.6 to 4.0 at 29 h postprandial and the fractions of W O2 consumption used by PVDO (percentage) were reduced (P less than .05) from 28.6 to 21.6 at 26 h and 25.2 to 18.2 at 27 h postprandial.(ABSTRACT TRUNCATED AT 250 WORDS)

Novel method for measuring growth of Treponema hyodysenteriae and its application for monitoring susceptibility of clinical isolates to antimicrobial agents.

A microdilution assay method was developed to determine MICs of antibiotics for Treponema hyodysenteriae and to estimate the bacterial burden of mice colonized by this swine pathogen. Hemolysis of bovine erythrocytes in broth was correlated with growth of the spirochete, and growth inhibition by drugs could be determined by measuring hemolysis. MICs of 10 antibiotics for eight clinical isolates of T. hyodysenteriae measured by this method were similar to values obtained by an agar dilution method and to values reported in the literature. This method was also used to demonstrate the elimination of T. hyodysenteriae from the ceca of infected mice following oral treatment with carbadox. The procedure described in this paper provides a rapid and simple method for determining MICs of antibiotics for T. hyodysenteriae and may be adapted for the purpose of determining the presence of this veterinary pathogen in cecal contents of experimentally infected mice.