Anti-mycotoxin feed additives: effects on metabolism, mycotoxin excretion, performance, and total tract digestibility of dairy cows fed artificially multi-mycotoxin-contaminated diets.

The aim of this study was to evaluate the effects of different anti-mycotoxin feed additives on the concentration of mycotoxins in milk, urine, and blood plasma of dairy cows fed artificially multi-mycotoxin-contaminated diets. Secondarily, performance, total-tract apparent digestibility of nutrients, and blood parameters were evaluated. Twelve multiparous cows (165 ± 45 d in milk, 557 ± 49 kg body weight, and 32.1 ± 4.57 kg/d milk yield at the start of the experiment) were blocked according to parity, milk yield, and days in milk and used in a 4 × 4 Latin square design experiment with 21-d periods, where the last 7 d were used for sampling and data analysis. Treatments were: 1) Mycotoxin group (MTX), basal diet (BD) without anti-mycotoxin feed additives; 2) Hydrated sodium calcium aluminosilicate (HSCA), HSCA added to the BD at 25g/cow/d; 3) Mycotoxin deactivator 15 (MD15), MD (Mycofix® Plus, dsm-firmenich) added to the BD at 15 g/cow/d; and 4) Mycotoxin deactivator 30 (MD30), MD added to the BD at 30 g/cow/d. Cows from all treatments were challenged with a blend of mycotoxins containing 404 µg aflatoxins B1 (AFB1), 5,025 µg deoxynivalenol (DON), 8,046 µg fumonisins (FUM), 195 µg T2 toxin (T2), and 2,034 µg of zearalenone (ZEN) added daily to the BD during the last 7 d of each period. Neither performance (milk yield and composition) nor nutrient digestibility was affected by treatments. All additives reduced aflatoxin M1 (AFM1) concentration in milk, whereas MD15 and MD30 group had lower excretion of AFM1 in milk than HSCA. DON, FUM, T2, or ZEN were not detected in milk of MD15 and MD30. Concentrations in milk of DON, FUM, T2, and ZEN were similar between MTX and HSCA. Except for AFM1, none of the analyzed mycotoxins were detected in urine of MD30 group. Comparing HSCA to MD treatments, the concentration of AFM1 was greater for HSCA, whereas MD30 was more efficient at reducing AFM1 in urine than MD15. AFM1, DON, FUM, and ZEN were not detected in the plasma of cows fed MD30, and DON was also not detected in MD15 group. Plasma concentration of FUM was lower for MD15, similar plasma FUM concentration was reported for HSCA and MTX. Plasma concentration of ZEN was lower for MD15 than MTX and HSCA. Serum concentrations of haptoglobin and hepatic enzymes were not affected by treatments. Blood concentration of sodium was lower in HSCA compared with MD15 and MD30 groups. In conclusion, the mycotoxin deactivator proved to be effective in reducing the secretion of mycotoxins in milk, urine, and blood plasma, regardless of the dosage. This reduction was achieved without adverse effects on milk production or total-tract digestibility in cows fed multi-mycotoxin-contaminated diets over a short-term period. Greater reductions in mycotoxin secretion were observed with full dose of MD.

Increasing doses of carbohydrases: Effects on rumen fermentation, nutrient digestibility, and performance of mid-lactation cows.

The objective of this study was to evaluate the effects of exogenous enzymes on nutrient intake and digestibility, rumen fermentation, and productivity of mid-lactating cows. Experiment 1 was designed to test increasing doses [0, 0.5, 1.0, or 1.5 g/kg of dry matter (DM)] of a combination of 2 enzyme products with xylanase and ß-glucanase activities (Ronozyme Wx and Ronozyme VP, respectively; DSM Nutritional Products) on rumen fermentation and total apparent digestibility. Enzyme combinations had a ratio of endo-1,3(4)-ß-glucanase to endo-1,4-ß-xylanase of 8:2 (wt/wt). For experiment 1, 8 rumen cannulated lactating cows were used into a double 4 × 4 Latin square design experiment with 14 d of diet adaptation and 7 d of sampling. Despite no differences in feed intake, carbohydrases linearly increased neutral detergent fiber digestibility. Treatments marginally affected rumen fermentation, where a linear trend for lower rumen pH and a linear trend for greater isobutyrate concentration were observed with increasing enzyme dose. A trend for lower rumen NH3-N concentration was observed for cows receiving carbohydrases in comparison with control group. When comparing all enzyme treatments against control group, cows fed enzymes tended to produce more 3.5% fat-corrected milk (FCM), produced more milk fat, and had greater blood glucose concentration. Experiment 2 evaluated 3 doses (0, 0.5, or 0.75 g/kg of DM) of the same combination of enzyme products on performance of cows (n = 36) in a complete randomized block (n = 12) design. Cows received treatments for 9 wk. No interaction effects between treatments and time were observed for all variables assessed in this study. In agreement with experiment 1, no differences were detected for feed intake, but cows fed the enzyme products tended to produce more 3.5% FCM and milk fat compared with control. In addition, cows fed enzymes exhibited greater efficiency of FCM production (FCM ÷ DM intake) compared with control. No differences were detected for intake and productivity when comparing the 2 doses of carbohydrases. In summary, the enzyme products tested in this study may improve feed efficiency due to greater milk fat concentration.

Nutrient digestibility, ruminal fermentation, and milk yield in dairy cows fed a blend of essential oils and amylase.

Two experiments were carried out to evaluate a blend of essential oils (EO) combined with amylase as an alternative to ionophores and its potential for reducing the use of antibiotics in the dairy industry. In experiment 1, 8 rumen-cannulated Holstein cows (576 ± 100 kg of body weight, 146 ± 35 d in milk, and 35.1 ± 4.0 kg/d of milk yield at the start of the experiment) were assigned to a 4 × 4 Latin square experiment with 21-d periods to determine the influence of feed additives on total apparent digestibility of nutrients, ruminal fermentation, N utilization, microbial protein synthesis, blood glucose and urea concentrations, and milk yield and composition in dairy cows. Treatment sequences assigned to cows in each block included no feed additives (control; CON); monensin (MON) added at 13 mg/kg of diet dry matter (DM); a blend of EO supplemented at 44 mg/kg of diet DM; and EO treatment combined with α-amylase at 330 kilo novo units/kg of diet DM (EOA). Differences among treatments were studied using orthogonal contrasts as follows: CON versus feed additives (MON, EO, and EOA), MON versus EO and EOA, and EO versus EOA. No differences were detected in nutrient intake and digestibility in cows. In general, feed additives decreased ruminal NH3-N concentration of cows, notably when diet was supplemented with MON. Furthermore, feed additives increased ruminal concentrations of acetate, butyrate, and branched-chain fatty acids. Cows fed treatments containing EO and EOA exhibited lower pH, higher NH3-N, and a trend to greater total volatile fatty acid concentration in the ruminal fluid compared with cows fed MON. Treatments containing EO increased ruminal butyrate concentration compared with MON. No treatment × time interaction effect was observed on ruminal fermentation measurements. Cows fed diets supplemented with feed additives had greater efficiency of N transfer into milk (milk N:N intake), whereas cows fed EOA exhibited greater N transfer into milk than those fed EO. Treatments had no effect on milk yield and composition, but feed additives increased the milk yield efficiency (milk yield divided by dry matter intake), whereas treatments containing EO had similar milk yield efficiency compared with MON. For experiment 2, 30 multiparous Holstein cows (574 ± 68 kg of body weight, 152 ± 54 d in milk, and 30.9 ± 4.1 kg/d of milk yield at the start of the experiment) were enrolled to a randomized complete block design experiment. The MON, EO, and EOA treatments were randomly assigned to cows within blocks (n = 10), and feed additives were provided throughout a 9-wk period. No differences were found in nutrient intake and digestibility, but cows fed EOA tended to exhibit greater dry matter intake than those fed EO. Blood metabolites and milk production were not affected by treatments. However, cows fed MON or EOA had greater milk protein content than those cows fed treatments containing EO. Feeding EO with or without amylase had similar response to feeding MON in terms of feed intake and milk yield, with a small negative effect on milk protein yield when feeding EO alone. Feed additives increased the concentrations of acetate, butyrate, and branched-fatty acids in ruminal fluid, whereas treatments containing EO had greater ruminal butyrate and NH3-N concentrations. Therefore, either EO or EOA can replace MON in diets of dairy cows while maintaining performance.

Effect of exogenous amylase on lactation performance of dairy cows fed a high-starch diet.

Exogenous amylase supplementation can increase starch and fiber digestibility in lactating dairy cows. We evaluated the effect of exogenous amylase supplementation on diets with high starch concentration (32% of dry matter). Twenty-eight Holstein cows (171 ± 80 d in milk, 4 primiparous) received a standard diet for 14 d and then a treatment for 63 d, in a covariate-adjusted randomized block design with repeated measures over time. Treatments were amylase [0.5 g of Ronozyme RumiStar (DSM Nutritional Products, Basel, Switzerland) per kg of total mixed ration dry matter] or control. The diets contained (% of dry matter): 39.4% corn silage, 11.2% rehydrated and ensiled mature corn grain, and 11.7% finely ground mature corn. Amylase increased milk yield (32.3 vs. 33.0 kg/d) and reduced dry matter intake (20.7 vs. 19.7 kg/d), increasing feed efficiency (1.52 vs. 1.63). Amylase also increased milk lactose synthesis (1.49 vs. 1.56 kg/d) and plasma glucose concentration (59.3 vs. 68.6 mg/dL). Secretions of milk fat and protein did not differ. Although milk urea N did not differ, amylase reduced the concentration of urea N in blood, suggesting an increase in ruminal starch degradation. However, the total-tract apparent digestibility of starch (96.3% of intake) and neutral detergent fiber (44.4% of intake), ruminal fermentation profile, and microbial yield estimated by urinary allantoin excretion did not differ. Cows fed amylase sorted in favor of long feed particles and against short particles, had shorter chewing activity (780 vs. 699 min/d), and had fewer meals per day (11.5 vs. 9.7). Amylase improved the feed efficiency of lactating cows fed a high-starch diet; the enzyme increased milk yield and reduced intake.

Effect of selected feed additives to improve growth and health of dairy calves.

The aim was to evaluate the effect of different feed additives on intake, performance, and fecal consistency index (FCI) of dairy calves from 6-60 d of age and its residual effect 15 d after weaning. Fifty Holstein calves (38 ± 1.0 kg BW) were fed 5 L/d of milk plus starter feed until weaning, and corn silage and concentrate after weaning. The treatments were: control (CON), monensin (MON; 30 mg/kg of starter), probiotic E. faecium (PROB; 70 mg/kg of starter), essential oils (EO; 300 mg/kg of starter), or PROB + EO (EOPROB). Fecal score and dry matter intake (DMI) were measured daily, and animals were weighed every 15 d. A DNA extraction from feces was performed to identify the presence of microorganisms (E. coli, Hafnia, Shiguella, Lactobacillus spp, Enterococcus spp, and Enterococcus faecium NCIMB 10415) by PCR. Two 72-h digestibility trials were performed at days 20-28 and 50-56, by total fecal collection. The DMI before weaning was greater for EO (903.0 g/d) compared with MON (794.3 g/d) and EOPROB (783.1 g/d). The FCI decreased during pre-weaning for EO and MON. Average daily gain (ADG) and feed efficiency (FE) did not differ among treatments before weaning. After weaning, DMI and FCI did not differ among treatments. The EO had greater ADG (917.5 g/d) compared with CON (615.8 g/d) and PROB (592.6 g/d). The FE improved with EO (0.72 g/g) over CON (0.36 g/g), MON (0.49 g/g), and PROB (0.36 g/g). The PCR results showed absence of E. faecium NCIMB 10415 in animals fed PROB and CON. Animals fed PROB had greater intake of CP and NDF than animals fed EOPROB. The EO can be added to the dairy calf ration to improve fecal score and increase DMI. The pre-weaning FCI decrease with MON and increase with PROB.

Minimum inhibitory concentrations of cephalosporin compounds and their active metabolites for selected mastitis pathogens.

OBJECTIVE: To compare the minimum inhibitory concentration (MIC) of cephapirin and ceftiofur with MICs of their active metabolites (desacetylcephapirin and desfuroylceftiofur) for selected mastitis pathogens. SAMPLE: 488 mastitis pathogen isolates from clinically and subclinically affected cows in commercial dairy herds in Wisconsin. PROCEDURES: Agar dilution was used to determine MICs for Staphylococcus aureus (n = 98), coagulase-negative staphylococci (99), Streptococcus dysgalactiae (97), Streptococcus uberis (96), and Escherichia coli (98). RESULTS: All S aureus isolates were susceptible to cephapirin and ceftiofur. Most coagulase-negative staphylococci were susceptible to cephapirin and ceftiofur. For E coli, 50 (51.0%; cephapirin) and 93 (94.95%; ceftiofur) isolates were susceptible to the parent compounds, but 88 (89.8%) were not inhibited at the maximum concentration of desacetylcephapirin. All S dysgalactiae isolates were susceptible to ceftiofur and cephapirin, and consistent MICs were obtained for all compounds. Most S uberis isolates were susceptible to cephapirin and ceftiofur. Of 98 S aureus isolates classified as susceptible to ceftiofur, 42 (42.9%) and 51 (52%) were categorized as intermediate or resistant to desfuroylceftiofur, respectively. For 99 coagulase-negative staphylococci classified as susceptible to ceftiofur, 45 (45.5%) and 17 (17.2%) isolates were categorized as intermediate or resistant to desfuroylceftiofur, respectively. For all staphylococci and streptococci, 100% agreement in cross-classified susceptibility outcomes was detected between cephapirin and desacetylcephapirin. No E coli isolates were classified as susceptible to desacetylcephapirin. CONCLUSIONS AND CLINICAL RELEVANCE: Differences in inhibition between parent compounds and their active metabolites may be responsible for some of the variation between clinical outcomes and results of in vitro susceptibility tests.

Risk factors associated with the antimicrobial resistance of Staphylococcus aureus isolated from bovine mastitis / Fatores de risco associados com a resistência antimicrobiana de Staphylococcus aureus isolados da mastite bovina

The objective of this study was to evaluate herd management practices and mastitis treatment procedures as risk factors associated with Staphylococcus aureus antimicrobial resistance. For this study, 13 herds were selected to participate in the study to evaluate the association between their management practices and mastitis treatment procedures and in vitro antimicrobial susceptibility. A total of 1069 composite milk samples were collected aseptically from the selected cows in four different periods over two years. The samples were used for microbiological culturing of S. aureus isolates and evaluation of their antimicrobial susceptibility. A total of 756 samples (70.7%) were culture-positive, and S. aureus comprised 27.77% (n=210) of the isolates. The S. aureus isolates were tested using the disk-diffusion susceptibility assay with the following antimicrobials: ampicillin 10mg; clindamycin 2μg; penicillin 1mg; ceftiofur 30μg; gentamicin 10mg; sulfa-trimethoprim 25μg; enrofloxacin 5μg; sulfonamide 300μg; tetracycline 30μg; oxacillin 1mg; cephalothin 30μg and erythromycin 5μg. The variables that were significantly associated with S. aureus resistance were as follows: the treatment of clinical mastitis for ampicillin (OR=2.18), dry cow treatment for enrofloxacin (OR=2.11) and not sending milk samples for microbiological culture and susceptibility tests, for ampicillin (OR=2.57) and penicillin (OR=4.69). In conclusion, the identification of risk factors for S. aureus resistance against various mastitis antimicrobials is an important information that may help in practical recommendations for prudent use of antimicrobial in milk production.(AU)

Objetivou-se com este estudo avaliar os fatores de risco associados às práticas de manejo e tratamento de mastite e a resistência aos antimicrobianos de Staphylococcus aureus isolados de vacas com mastite. Foram selecionados para o presente estudo 13 rebanhos localizados na região de Pirassununga/SP. Foi aplicado um questionário contendo informações para o levantamento de fatores de risco relacionados à resistência aos antimicrobianos e às práticas de manejo e tratamento de mastite. Após a seleção dos rebanhos e aplicação dos questionários, foram utilizados 210 isolados de S. aureus de amostras compostas de leite coletadas durante 24 meses, em quatro períodos, para realização dos testes de resistência. Os antimicrobianos testados foram: ampicilina 10µg, clindamicina 2µg, penicilina 1µg, eftiofour 30µg, gentamicina 10µg, sulfatrimetropin 25µg, enrofloxacina 5µg, sulfonamida 300µg, tetraciclina 30µg, oxacilina 1µg, cefalotina 30µg e eritromicina 5µg. As variáveis que foram significativamente associadas à resistência de S. aureus foram: o tratamento da mastite clínica para ampicilina (OR = 2,18), o tratamento da vaca seca para enrofloxacina (OR=2,11), e o não envio de amostras de leite para a cultura microbiológica e testes de sensibilidade, para ampicilina (OR=2,57) e penicilina (OR=4,69). Em conclusão, a identificação dos fatores de risco para a resistência S. aureus frente aos principais agentes antimicrobianos, utilizados para tratamento da mastite, pode auxiliar o estabelecimento do uso prudente de antimicrobianos na produção de leite.(AU)

Risk factors associated with the antimicrobial resistance of Staphylococcus aureus isolated from bovine mastitis / Fatores de risco associados com a resistência antimicrobiana de Staphylococcus aureus isolados da mastite bovina

The objective of this study was to evaluate herd management practices and mastitis treatment procedures as risk factors associated with Staphylococcus aureus antimicrobial resistance. For this study, 13 herds were selected to participate in the study to evaluate the association between their management practices and mastitis treatment procedures and in vitro antimicrobial susceptibility. A total of 1069 composite milk samples were collected aseptically from the selected cows in four different periods over two years. The samples were used for microbiological culturing of S. aureus isolates and evaluation of their antimicrobial susceptibility. A total of 756 samples (70.7%) were culture-positive, and S. aureus comprised 27.77% (n=210) of the isolates. The S. aureus isolates were tested using the disk-diffusion susceptibility assay with the following antimicrobials: ampicillin 10mg; clindamycin 2μg; penicillin 1mg; ceftiofur 30μg; gentamicin 10mg; sulfa-trimethoprim 25μg; enrofloxacin 5μg; sulfonamide 300μg; tetracycline 30μg; oxacillin 1mg; cephalothin 30μg and erythromycin 5μg. The variables that were significantly associated with S. aureus resistance were as follows: the treatment of clinical mastitis for ampicillin (OR=2.18), dry cow treatment for enrofloxacin (OR=2.11) and not sending milk samples for microbiological culture and susceptibility tests, for ampicillin (OR=2.57) and penicillin (OR=4.69). In conclusion, the identification of risk factors for S. aureus resistance against various mastitis antimicrobials is an important information that may help in practical recommendations for prudent use of antimicrobial in milk production.

Objetivou-se com este estudo avaliar os fatores de risco associados às práticas de manejo e tratamento de mastite e a resistência aos antimicrobianos de Staphylococcus aureus isolados de vacas com mastite. Foram selecionados para o presente estudo 13 rebanhos localizados na região de Pirassununga/SP. Foi aplicado um questionário contendo informações para o levantamento de fatores de risco relacionados à resistência aos antimicrobianos e às práticas de manejo e tratamento de mastite. Após a seleção dos rebanhos e aplicação dos questionários, foram utilizados 210 isolados de S. aureus de amostras compostas de leite coletadas durante 24 meses, em quatro períodos, para realização dos testes de resistência. Os antimicrobianos testados foram: ampicilina 10µg, clindamicina 2µg, penicilina 1µg, eftiofour 30µg, gentamicina 10µg, sulfatrimetropin 25µg, enrofloxacina 5µg, sulfonamida 300µg, tetraciclina 30µg, oxacilina 1µg, cefalotina 30µg e eritromicina 5µg. As variáveis que foram significativamente associadas à resistência de S. aureus foram: o tratamento da mastite clínica para ampicilina (OR = 2,18), o tratamento da vaca seca para enrofloxacina (OR=2,11), e o não envio de amostras de leite para a cultura microbiológica e testes de sensibilidade, para ampicilina (OR=2,57) e penicilina (OR=4,69). Em conclusão, a identificação dos fatores de risco para a resistência S. aureus frente aos principais agentes antimicrobianos, utilizados para tratamento da mastite, pode auxiliar o estabelecimento do uso prudente de antimicrobianos na produção de leite.