Comparative genomics of Enterococcus spp. isolated from bovine feces.

BACKGROUND: Enterococcus is ubiquitous in nature and is a commensal of both the bovine and human gastrointestinal (GI) tract. It is also associated with clinical infections in humans. Subtherapeutic administration of antibiotics to cattle selects for antibiotic resistant enterococci in the bovine GI tract. Antibiotic resistance genes (ARGs) may be present in enterococci following antibiotic use in cattle. If located on mobile genetic elements (MGEs) their dissemination between Enterococcus species and to pathogenic bacteria may be promoted, reducing the efficacy of antibiotics. RESULTS: We present a comparative genomic analysis of twenty-one Enterococcus spp. isolated from bovine feces including Enterococcus hirae (n = 10), Enterococcus faecium (n = 3), Enterococcus villorum (n = 2), Enterococcus casseliflavus (n = 2), Enterococcus faecalis (n = 1), Enterococcus durans (n = 1), Enterococcus gallinarum (n = 1) and Enterococcus thailandicus (n = 1). The analysis revealed E. faecium and E. faecalis from bovine feces share features with human clinical isolates, including virulence factors. The Tn917 transposon conferring macrolide-lincosamide-streptogramin B resistance was identified in both E. faecium and E. hirae, suggesting dissemination of ARGs on MGEs may occur in the bovine GI tract. An E. faecium isolate was also identified with two integrative conjugative elements (ICEs) belonging to the Tn916 family of ICE, Tn916 and Tn5801, both conferring tetracycline resistance. CONCLUSIONS: This study confirms the presence of enterococci in the bovine GI tract possessing ARGs on MGEs, but the predominant species in cattle, E. hirae is not commonly associated with infections in humans. Analysis using additional complete genomes of E. faecium from the NCBI database demonstrated differential clustering of commensal and clinical isolates, suggesting that these strains may be specifically adapted to their respective environments.

Dose-response effects of dietary pequi oil on fermentation characteristics and microbial population using a rumen simulation technique (Rusitec).

The effect of increasing the concentration of commercial pequi (Caryocar brasiliense) oil on fermentation characteristics and abundance of methanogens and fibrolityc bacteria was evaluated using the rumen simulation technique (Rusitec). In vitro incubation was performed over 15 days using a basal diet consisting of ryegrass, maize silage and concentrate in equal proportions. Treatments consisted of control diet (no pequi oil inclusion, 0 g/kg DM), pequi dose 1 (45 g/kg DM), and pequi dose 2 (91 g/kg DM). After a 7 day adaptation period, samples for fermentation parameters (total gas, methane, and VFA production) were taken on a daily basis. Quantitative real time PCR (q-PCR) was used to evaluate the abundance of the main rumen cellulolytic bacteria, as well as abundance of methanogens. Supplementation with pequi oil did not reduce overall methane production (P = 0.97), however a tendency (P = 0.06) to decrease proportion of methane in overall microbial gas was observed. Increasing addition of pequi oil was associated with a linear decrease (P < 0.01) in dry matter disappearance of maize silage. The abundance of total methanogens was unchanged by the addition of pequi oil, but numbers of those belonging to Methanomassiliicoccaceae decreased in liquid-associated microbes (LAM) samples (P < 0.01) and solid-associated microbes (SAM) samples (P = 0.09) respectively, while Methanobrevibacter spp. increased (P < 0.01) only in SAM samples. Fibrobacter succinogenes decreased (P < 0.01) in both LAM and SAM samples when substrates were supplemented with pequi oil. In conclusion, pequi oil was ineffective in mitigating methane emissions and had some adverse effects on digestibility and selected fibrolytic bacteria.

Inclusion of glycerol in forage diets increases methane production in a rumen simulation technique system.

We hypothesised that the inclusion of glycerol in the forage diets of ruminants would increase the proportion of propionate produced and thereby decrease in vitro CH4 production. This hypothesis was examined in the present study using a semi-continuous fermentation system (rumen simulation technique) fed a brome hay (8·5 g) and maize silage (1·5 g) diet with increasing concentrations (0, 50, 100 and 150 g/kg DM) of glycerol substituted for maize silage. Glycerol linearly increased total volatile fatty acids production (P<0·001). Acetate production was quadratically affected (P=0·023) and propionate and butyrate production was linearly increased (P<0·001). Glycerol linearly increased (P=0·011) DM disappearance from hay and silage. Crude protein disappearance from hay was not affected (P=0·789), but that from silage was linearly increased (P<0·001) with increasing glycerol concentrations. Neutral-detergent fibre (P=0·040) and acid-detergent fibre (P=0·031) disappearance from hay and silage was linearly increased by glycerol. Total gas production tended to increase linearly (P=0·061) and CH4 concentration in gas was linearly increased (P<0·001) by glycerol, resulting in a linear increase (P<0·001) in mg CH4/g DM digested. Our hypothesis was rejected as increasing concentrations of glycerol in a forage diet linearly increased CH4 production in semi-continuous fermenters, despite the increases in the concentrations of propionate. In conclusion, this apparent discrepancy is due to the more reduced state of glycerol when compared with carbohydrates, which implies that there is no net incorporation of electrons when glycerol is metabolised to propionate.

Partial replacement of an artificial nectar diet with native browse for feather-tail gliders (Acrobates pygmaeus) in captivity.

Captive-bred feather-tail gliders (Acrobates pygmaeus) housed at Taronga Zoo have had a long history of eye cholesterol plaques that may be associated with a largely sugar-based diet such as artificial nectar. The gliders also have prolonged periods of reduced activity when they are not visible in exhibits. This may be due to the ad libitum supply of an energy rich feed and reduced need to forage. This study examined behavioral and physiological changes associated with supplementing the high sugar-based diet with two species of native browse. The experiment was conducted over two consecutive periods of 3 weeks and consisted of two treatment groups: one group was offered the artificial nectar only, while the other group was offered the artificial nectar supplemented with a variety of native flowers. Live weight was recorded weekly. There was no change (P > 0.10) in artificial nectar intake with the supplementation of native browse in the diet. Blood metabolites (cholesterol, triglycerides, glucose) tested for the two groups had no differences (P > 0.10) between treatments. Upon examination, there were no signs of tooth decay or cholesterol plaques in all animals throughout the experiment. Feed intake and behavior were recorded via sensor cameras. There was an increase (P < 0.05) in the daily foraging activity of gliders supplemented with native flowers compared to gliders fed the artificial nectar alone. In conclusion, supplementing to provide a more native diet to A. pygmaeus enhanced their natural foraging behavior, suggesting that it may result in long-term improvements in their health.

Effects of cinnamon leaf, oregano and sweet orange essential oils on fermentation and aerobic stability of barley silage.

BACKGROUND: Silage additives are marketed with the primary aim of improving the fermentation and/or aerobic stability of silage. The objective of this study was to evaluate the impact of three different essential oils (EOs; cinnamon leaf (CIN), oregano (ORE) and sweet orange (SO)) on the fermentation characteristics and stability of barley silage. Chopped whole-plant barley (Hordem vulgare L.) forage was ensiled either untreated (0 mg kg⁻¹ dry matter (DM)) or treated with CIN, ORE or SO (37.5, 75 and 120 mg kg⁻¹ DM). RESULTS: Moulds were not detected in any treatments, including the control, after 7 days of air exposure. All EOs at a concentration of 120 mg kg⁻¹ silage DM decreased (P = 0.001) yeast populations in comparison with the control during air exposure. Net gas, methane and ammonia concentrations in vitro did not differ among treatments. Changes in volatile fatty acid concentrations were small, and in situ data showed no changes in DM and neutral detergent fibre digestion rates for CIN, ORE or SO at concentrations up to 120 mg kg⁻¹ DM. CONCLUSION: The findings from this study show that a concentration of 120 mg EO kg⁻¹ DM decreased yeast counts during aerobic stability tests. However, all EO treatments had minimal effects on data from in vitro and in situ incubations.

Fermentation characteristics of corn-, triticale-, and wheat-based dried distillers' grains with solubles in barley-based diets determined using continuous and batch culture systems.

BACKGROUND: Dried distillers' grains with solubles (DDGS) are becoming increasingly available and popular as an alternative livestock feed. This study used continuous and batch culture techniques to compare the in vitro fermentation characteristics of diets containing corn-, triticale- or wheat-based DDGS at 200 g kg(-1) diet dry matter (DM) against a barley grain-based control diet. RESULTS: In continuous fermentation of wheat DDGS diet, total volatile fatty acid (VFA) concentration was decreased by 15.7% across sampling times and NH(3) concentration was quadrupled compared with control, whereas fermentations of corn- and triticale-DDGS diets were similar to the barley-based control. In batch cultures, corn DDGS differed from control only in increased culture pH. Compared with control diet, triticale DDGS yielded lower total gas production (140.94 versus 148.78 mL g(-1) DM) and in vitro DM digestibility (IVDMD; 0.509 versus 0.535). Wheat DDGS diet yielded decreased total gas production (123.49 mL g(-1) DM) and IVDMD (0.468), as well as total VFA production (105.84 versus 134.20 mmol L(-1)) and substantially increased ammonia concentration (151.61 versus 58.34 mg L(-1)) and acetate:propionate ratio (2.94 versus 1.11). CONCLUSION: Corn- and triticale- DDGS diets exhibited fermentation characteristics similar to the barley based control diet, consistent with in vivo findings that these diets yielded no adverse effects on production. In vitro ruminal fermentation of wheat DDGS diet differed significantly from control in several aspects including 2.6 to 6X higher ammonia concentrations.

Humic Substances Alter Ammonia Production and the Microbial Populations Within a RUSITEC Fed a Mixed Hay - Concentrate Diet.

Humic substances are a novel feed additive which may have the potential to mitigate enteric methane (CH4) production from ruminants as well as enhance microbial activity in the rumen. The aim of this study was to examine the effects of humic substances on fermentation characteristics and microbial communities using the rumen stimulation technique (RUSITEC). The experiment was conducted as a completely randomized design with 3 treatments duplicated in 2 runs (a 15-day period each run) with 2 replicates per run. Treatments consisted of a control diet (forage:concentrate; 60:40) without humic substances or humic substances added at either 1.5 g/d or 3.0 g/d. Dry matter disappearance, pH, fermentation parameters and gas production were measured from day 8 to 15. Samples for microbial profiling were taken on day 5, 10, and 15 using the digested feed bags for solid- associated microbes (SAM) and fermenter fluid for liquid- associated microbes (LAM). The inclusion of humic substances had no effect (P ≥ 0.19) on DM disappearance, pH or the concentrations of VFA. The production of NH3 was linearly decreased (P = 0.04) with increasing levels of humic substances in the diet. There was no effect (P ≥ 0.43) of humic substances on total gas, CO2 or CH4 production. The number of OTUs was significantly reduced in the 3.0 g/d treatment compared to the control on d 10 and 15; however, the microbial community structure was largely unaffected (P > 0.05). In the SAM samples, the genera Lachnospiraceae XPB1014 group, Succiniclasticum, and Fibrobacter were reduced in the 3.0 g/d treatment and Anaeroplasma, Olsenella, and Pseudobutyrivibrio were increased on day 5, 10, and 15. Within the LAM samples, Christensenellaceae R-7 and Succiniclasticum were the most differentially abundant genera between the control and 3.0 g/d HS treatment samples (P < 0.05). This study highlights the potential use of humic substances as a natural feed additive which may play a role in nitrogen metabolism without negatively affecting the ruminal microbiota.

Tucumã Oil Shifted Ruminal Fermentation, Reducing Methane Production and Altering the Microbiome but Decreased Substrate Digestibility Within a RUSITEC Fed a Mixed Hay - Concentrate Diet.

Tucumã oil is sourced from the fruit pulp of the tucumã tree and contains high concentrations of unsaturated fatty acids and carotenoids. Due to these properties it may have the potential to decrease enteric methane (CH4) from ruminants when included in the diet. The objective of this study was to determine the effect of oil mechanically extracted from the fruit pulp of tucumã on fermentation characteristics, CH4 production and the microbial community using the rumen stimulation technique. Treatments consisted of a control diet (forage:concentrate; 70:30), and tucumã oil included at 0.5 or 1.0% (v/v). Addition of tucumã oil linearly decreased (P < 0.01) dry matter disappearance. Total gas (mL/d) and carbon dioxide (CO2) production (mL/d, mL/g DM) were unaffected (P ≥ 0.36) to increasing addition of tucumã oil where 0.5% (v/v) of Tucumã oil numerically increased both variables. Acetate and butyrate percentages of total VFA were linearly decreased (P ≤ 0.01) and propionate and valerate percentages of total VFA were linearly increased (P < 0.01) by increasing concentrations of tucumã oil added to the substrate. The ratio of acetate to propionate was linearly decreased (P < 0.01) with increasing concentration of tucumã oil. Methane production (mL/d) was linearly decreased (P = 0.04) with increasing addition of tucumã oil to the substrate. Tucumã oil reduced the bacterial richness and diversity when included at 1.0% (v/v) in both solid- and liquid- associated microbes. The abundance of the genera Fibrobacter and Rikenellaceae RC9 gut group were decreased and Pyramidobacter, Megasphaera, Anaerovibrio, and Selenomonas were enriched by the addition of 1.0% tucumã oil. In conclusion, tucumã oil resulted in the favorable shift in fermentation products away from acetate toward propionate, decreasing the production of CH4 when tucumã oil was included at 1.0% (v/v), however, substrate digestibility was also inhibited. The rumen microbiota was also altered by the addition of tucumã oil.

Comparison of antimicrobial resistance genes in feedlots and urban wastewater.

The use of antibiotics in livestock production in North America and possible association with elevated abundance of detectable antimicrobial resistance genes (ARG) is a growing concern. Real-time, quantitative PCR (RT-qPCR) was used to determine the relative abundance and diversity of ARG in fecal composite and catch basin samples from 4 beef feedlots in Alberta. Samples from a surrounding waterway and municipal wastewater treatment plants were also included to compare the ARG profile of urban environments and fresh water with that of feedlots. The relative abundance of 18 resistance genes across 5 antibiotic families including sulfonamides, tetracyclines, macrolides, fluoroquinolones, and ß-lactams was examined. Sulfonamide, fluoroquinolone, and ß-lactam resistance genes predominated in wastewater treatment samples, while tetracycline resistance genes predominated in cattle fecal composite samples. These results reflect the types of antibiotic that are used in cattle versus humans, but other factors such as co-selection of ARG and variation in the composition of bacterial communities associated with these samples may also play a role.

En Amérique du Nord, l'utilisation des antibiotiques dans la production du bétail et l'association possible avec une abondance élevée détectable de gènes de résistance aux antimicrobiens (GRA) est une préoccupation grandissante. Une épreuve d'amplification en chaîne par la polymérase quantitative en temps réel a été utilisée afin de déterminer l'abondance relative et la diversité des GRA dans des échantillons composites de fèces et de bassin de rétention de quatre parcs d'engraissement de bovins en Alberta. Des échantillons d'un cours d'eau avoisinant et de l'usine municipale de traitement des eaux usées ont également été inclus afin de comparer le profil des GRA provenant d'un milieu urbain et d'eau fraîche à celui des parcs d'engraissement. L'abondance relative de 18 gènes de résistance issus de cinq familles d'antibiotiques incluant les sulfonamides, les tétracyclines, les macrolides, les fluoroquinolones, et les ß-lactames fut examinée. Les gènes de résistance aux sulfonamides, aux fluoroqunolones, et aux ß-lactames prédominaient dans les échantillons d'eaux usées, alors que les gènes de résistance à la tétracycline étaient prédominants dans les échantillons composites de fèces des bovins. Ces résultats reflètent les types d'antibiotiques qui sont utilisés chez les bovins versus les humains, mais d'autres facteurs tels que la co-sélection de GRA et la variation dans la composition des communautés bactériennes associées à ces échantillons peuvent également jouer un rôle.(Traduit par Docteur Serge Messier).

Unconventional Vegetable Oils for a Reduction of Methanogenesis and Modulation of Ruminal Fermentation.

The objective of this study was to evaluate the use of vegetable oils from plants grown in Brazil, first using the in vitro batch culture, and then evaluating the oil with methane (CH4) reducing potential in an in vivo experiment. The in vitro experiment was conducted as a completely randomized design using the seven contrasting oils. Treatments consisted of a control and 3 increasing concentrations (0, 1, 2, and 5% v/v) of oil added to a tifton 85 hay samples. All vegetable oils linearly decreased (P < 0.01) gas production after 24 h of incubation, with the greatest reduction when 5% of oil was included into the diet. Açaí and buriti had no effect of CH4 (% or mL/g DM incubated) however carrot, macaúba, basil, passionflower, and pequi oil all linearly decreased (P < 0.01) CH4 production with increasing inclusion rate of oil. Pequi oil resulted in the largest decrease in CH4 production (mL/g DM incubated) after 24 h of in vitro incubation. The objective of the in vivo experiment was to evaluate the effects of pequi oil on nutrient digestibility, CH4 production, and rumen fermentation parameters in wethers fed a hay-based diet. The experiment was conducted as a 2 × 2 Latin Square design using 4 Dorper wethers (63.4 ± 1.46 kg body weight). There were 2 experimental periods of 21 d each, with d 1-14 used for diet adaptation and d 15-21 for measurements and collections. The treatments consisted of a control diet and pequi oil fed at 70 g per animal per day. The addition of pequi oil to the diet had no effect on feed intake or the digestibility of nutrients, however there was a numerical decrease in the population of cellulolytic bacteria. There was a tendency (P = 0.06) for pequi oil addition to decrease CH4 production (g/d) by 17.5%. From this study, we can conclude that pequi oil may be used as a suitable oil for reducing CH4 production from ruminants, with no negative effects on intake or digestibility.

The Type of Forage Substrate Preparation Included as Substrate in a RUSITEC System Affects the Ruminal Microbiota and Fermentation Characteristics.

In vitro fermentation systems such as the rumen simulation technique (RUSITEC) are frequently used to assess dietary manipulations in livestock, thereby limiting the use of live animals. Despite being in use for nearly 40 years, improvements are continually sought in these systems to better reflect and mimic natural processes in ruminants. The aim of this study was to evaluate the effect of forage preparation, i.e., frozen minced (FM) and freeze-dried and ground (FDG), on the ruminal microbiota and on fermentation characteristics when included as a substrate in a RUSITEC system. A completely randomized design experiment was performed over a 15-day period, with 7 days of adaptation and an 8-day experimental period. Fermentation parameters (total gas, CH4, and volatile fatty acid production) were analyzed on a daily basis over the experimental period and the archaeal and bacterial microbiota (liquid-associated microbes [LAM] and solid-associated microbes [SAM] was assessed at 0, 5, 10, and 15 days using high-throughput sequencing of the 16S rRNA gene. Results from this study suggested a tendency (P = 0.09) of FM treatment to increase daily CH4 (mg/d) production by 16.7% when compared with FDG treatment. Of the major volatile fatty acids (acetate, propionate, and butyrate), only butyrate production was greater (P = 0.01) with FM treatment compared with FDG substrate. The archaeal and bacterial diversity and richness did not differ between the forage preparations, although feed particle size of the forage had a significant effect on microbial community structure in the SAM and LAM samples. The Bacteroidetes phylum was more relatively abundant in the FM substrate treatment, while Proteobacteria was enriched in the FDG treatment. At the genus-level, Butyrivibrio, Prevotella, and Roseburia were enriched in the FM substrate treatment and Campylobacter and Lactobacillus in the FDG substrate treatment. Evidence from this study suggests that forage preparation affects CH4 production, butyrate production, and the structure of the rumen microbiota during in vitro fermentation.

Incubation Temperature, But Not Pequi Oil Supplementation, Affects Methane Production, and the Ruminal Microbiota in a Rumen Simulation Technique (Rusitec) System.

Lipid supplementation is a promising strategy for methane mitigation in cattle and has been evaluated using several different lipid sources. However, limited studies have assessed the effect of temperature on methane emissions from cattle and changes in incubation temperature have also not been extensively evaluated. The aim of this study was to evaluate the combined effect of pequi oil (high in unsaturated fatty acids) and incubation temperature on fermentation characteristics and microbial communities using the rumen simulation technique. A completely randomized experiment was conducted over a 28-day period using a Rusitec system. The experiment was divided into four periods of 7 days each, the first of which was a 7-day adaptation period followed by three experimental periods. The two treatments consisted of a control diet (no pequi oil inclusion) and a diet supplemented with pequi oil (1.5 mL/day) which increased the dietary fat content to 6% (dry matter, DM-basis). Three fermenter vessels (i.e., replicates) were allocated to each treatment. In the first experimental period, the incubation temperature was maintained at 39°C, decreased to 35°C in the second experimental period and then increased again to 39°C in the third. Pequi oil was continuously supplemented during the experiment. Microbial communities were assessed using high-throughput sequencing of the archaeal and bacterial 16S rRNA gene. Methane production was reduced by 57% following a 4°C decrease in incubation temperature. Supplementation with pequi oil increased the dietary fat content to 6% (DM-basis) but did not affect methane production. Analysis of the microbiota revealed that decreasing incubation temperature to 35°C affected the archaeal and bacterial diversity and richness of liquid-associated microbes, but lipid supplementation did not change microbial diversity.

Effects of Continuously Feeding Diets Containing Cereal Ergot Alkaloids on Nutrient Digestibility, Alkaloid Recovery in Feces, and Performance Traits of Ram Lambs.

Allowable limits for cereal ergot alkaloids in livestock feeds are being re-examined, and the objective of this study was to compare nutrient digestibility, growth performance and carcass characteristics of ram lambs fed a range of alkaloid concentrations, including the maximum currently allowed in Canada (2 to 3 ppm). Four pelleted diets were fed: control, with no added alkaloids; 930; 1402; and 2447 ppb alkaloids based on total R and S epimers. Eight ram lambs (30.0 ± 3.1 kg) were used to examine the impacts of dietary treatments on nutrient digestibility and alkaloid recovery from feces. Concentrations of dietary alkaloids evaluated did not affect nutrient digestibility or N metabolism. Excepting ergocornine and ergocryptine, recovery of alkaloids in feces varied among periods, suggesting that individual lambs may differ in their ability to metabolize ergocristine, ergometrine, ergosine, ergotamine and their S epimers. In a second experiment, ram lambs (n = 47, 30 ± 8 kg) were randomly assigned to a diet and weighed weekly until they achieved a slaughter weight of ≥ 45 kg (average 9 weeks; range 6 to 13 weeks). Intake of DM did not differ (p = 0.91) among diets, although lambs fed 2447 ppb alkaloids had a lower (p < 0.01) ADG than did lambs receiving other treatments. The concentration of serum prolactin linearly declined (p < 0.01) with increasing alkaloids. Feeding 2447 ppb total alkaloids negatively impacted growth, while feeding 1402 ppb did not harm growth performance, but reduced carcass dressing percentage. Due to different concentrations of alkaloids affecting growth and carcass characteristics in the present study, determining allowable limits for total dietary alkaloids will require a better understanding of impacts of alkaloid profiles and interactions among individual alkaloids.

Draft Genome Sequence of an Enterococcus thailandicus Strain Isolated from Bovine Feces.

Here, we report the first draft genome sequence of Enterococcus thailandicus isolated from the feces of feedlot cattle in Southern Alberta.

Effect of in-feed administration and withdrawal of tylosin phosphate on antibiotic resistance in enterococci isolated from feedlot steers.

Tylosin phosphate is a macrolide commonly administered to cattle in North America for the control of liver abscesses. This study investigated the effect of in-feed administration of tylosin phosphate to cattle at subtherapeutic levels and its subsequent withdrawal on macrolide resistance using enterococci as an indicator bacterium. Fecal samples were collected from steers that received no antibiotics and steers administered tylosin phosphate (11 ppm) in-feed for 197 days and withdrawn 28 days before slaughter. Enterococcus species isolated from fecal samples were identified through sequencing the groES-EL intergenic spacer region and subject to antimicrobial susceptibility testing, identification of resistance determinants and pulsed-field gel electrophoresis profiling. Tylosin increased (P < 0.05) the proportion of ery(R) and tyl(R) enterococci within the population. Just prior to its removal, the proportion of ery(R) and tyl(R) resistant enterococci began decreasing and continued to decrease after tylosin was withdrawn from the diet until there was no difference (P > 0.05) between treatments on d 225. This suggests that antibiotic withdrawal prior to slaughter contributes to a reduction in the proportion of macrolide resistant enterococci entering the food chain. Among the 504 enterococci isolates characterized, Enterococcus hirae was found to predominate (n = 431), followed by Enterococcus villorum (n = 32), Enterococcus faecium (n = 21), Enterococcus durans (n = 7), Enterococcus casseliflavus (n = 4), Enterococcus mundtii (n = 4), Enterococcus gallinarum (n = 3), Enterococcus faecalis (n = 1), and Enterococcus thailandicus (n = 1). The diversity of enterococci was greater in steers at arrival than at exit from the feedlot. Erythromycin resistant isolates harbored the erm(B) and/or msrC gene. Similar PFGE profiles of ery(R) E. hirae pre- and post-antibiotic treatment suggest that increased abundance of ery(R) enterococci after administration of tylosin phosphate reflects selection for strains that were already present within the gastrointestinal tract of cattle at arrival.