Comparison of antibiotic resistance of udder pathogens in dairy cows kept on organic and on conventional farms

There has been a rapid rise in the emergence of multi-drug-resistant pathogens in the past 10 to 15 yr and some bacteria are now resistant to most antimicrobial agents. Antibiotic use is very restricted on Swiss organic dairy farms, and a purely prophylactic use, such as for dry cow mastitis prevention, is forbidden. A low prevalence of antibiotic resistance in organic farms can be expected compared with conventional farms because the bacteria are infrequently or not exposed to antibiotics. The occurrence of antibiotic resistance was compared between mastitis pathogens (Staphylococcus aureus, nonaureus staphylococci, Streptococcus dysgalactiae, Streptococcus uberis) from farms with organic and conventional dairy production. Clear differences in the percentage of antibiotic resistance were mainly species-related, but did not differ significantly between isolates from cows kept on organic and conventional farms, except for Streptococcus uberis, which exhibited significantly more single resistances (compared with no resistance) when isolated from cows kept on organic farms (6/10 isolates) than on conventional farms (0/5 isolates). Different percentages were found (albeit not statistically significant) in resistance to ceftiofur, erythromycin, clindamycin, enrofloxacin, chloramphenicol, penicillin, oxacillin, gentamicin, tetracycline, and quinupristin-dalfopristin, but, importantly, none of the strains was resistant to amoxicillin-clavulanic acid or vancomycin. Multidrug resistance was rarely encountered. The frequency of antibiotic resistance in organic farms, in which the use of antibiotics must be very restricted, was not different from conventional farms, and was contrary to expectation. The antibiotic resistance status needs to be monitored in organic farms as well as conventional farms and production factors related to the absence of reduced antibiotic resistance in organic farms need to be evaluated.     

Production effects and bioavailability of N-acetyl-l-methionine in lactating dairy cows

Two experiments were conducted to investigate the production effects of N-acetyl-l-methionine (NALM; experiment 1) and to estimate its bioavailability (BA) and rumen escape (RE; experiment 2), respectively, in lactating dairy cows. In experiment 1, 18 multiparous Holstein cows were used in a replicated, 3 × 3 Latin square design experiment with three 28-d periods. Treatments were (1) basal diet estimated to supply 45 g/d digestible Met (dMet) or 1.47% of metabolizable protein (MP; control), (2) basal diet top-dressed with 32 g/d of NALM to achieve dMet supply of 2.2% of MP, and (3) basal diet top-dressed with 56 g/d of NALM to achieve dMet supply of 2.6% of MP. The NALM treatments supplied estimated 17 and 29 g/d dMet from NALM, respectively, based on manufacturer's specifications. In experiment 2, 4 rumen-cannulated lactating Holstein cows were used in a 4 × 4 Latin square design experiment with four 12-d periods. A 12-d period for baseline data collection and 4 d for determination of RE of NALM preceded the Latin square experiment. For determination of RE, 30 g of NALM were dosed into the rumen simultaneously with Cr-EDTA (used as a rumen fluid kinetics marker) and samples of ruminal contents were collected at 0 (before dosing), 1, 2, 4, 6, 8, 10, 14, 18, and 24 h after dosing. Rumen escape of NALM was calculated using the estimated passage rate based on the measured Cr rate of disappearance. Bioavailability of abomasally dosed NALM was determined using the area under the curve of plasma Met concentration technique. Two doses of l-Met (providing 7.5 and 15 g of dMet) and 2 doses of NALM (11.2 and 14.4 g of dMet) were separately pulse-dosed into the abomasum of the cows and blood was collected from the jugular vein for Met concentration analysis at 0 (before dosing), 1, 2, 4, 6, 8, 10, 12, 14, 18, and 24 h after dosing. Supplementation of NALM did not affect DMI, milk yield, feed efficiency, or milk protein and lactose concentrations and yields in experiment 1. Milk fat concentration and energy-corrected milk yield decreased linearly with NALM dose. Plasma Met concentration was not affected by NALM dose. The estimated relative BA of abomasally dosed NALM (experiment 2) was 50% when dosed at 14.4 g/cow (11.2 g/d dMet from NALM) and 24% when dosed at 28.8 g/cow (14.4 g/d dMet from NALM). The estimated RE of NALM was 19% based on the measured k_p of Cr at 11%/h. The total availability of ingested NALM was estimated at 9.5% for the lower NALM dose when taking into account RE (19%) and bioavailability in the small intestine (50%). Overall, NALM supplementation to mid-lactation dairy cows fed a MP-adequate basal diet below NRC (2001) recommendations (45 g/d or 1.47% Met of MP) decreased milk fat and energy-corrected milk yields but did not affect milk or milk true protein yields. Further evaluation of BA of NALM at different doses is warranted. In addition, intestinal conversion of NALM to Met needs additional investigation to establish a possible saturation of the enzyme aminoacylase I at higher NALM doses.     

Quantifying body water kinetics and fecal and urinary water output from lactating Holstein dairy cows

Reliable estimates of fresh manure water output from dairy cows help to improve storage design, enhance efficiency of land application, quantify the water footprint, and predict nutrient transformations during manure storage. The objective of the study was to construct a mechanistic, dynamic, and deterministic mathematical model to quantify urinary and fecal water outputs (kg/d) from individual lactating dairy cows. The model contained 4 body water pools: reticulorumen (Q_RR), post-reticulorumen (Q_PR), extracellular (Q_EC), and intracellular (Q_IC). Dry matter (DM) intake, dietary forage, DM, crude protein, acid detergent fiber and ash contents, milk yield, and milk fat and protein contents, days in milk, and body weight were input variables to the model. A set of linear equations was constructed to determine drinking, feed, and saliva water inputs to Q_RR and fractional water passage from Q_RR to Q_PR. Water transfer via the rumen wall was subjected to changes in Q_EC and total water input to Q_RR. Post-reticulorumen water passage was adjusted for DM intake. Metabolic water production and respiratory cutaneous water losses were estimated with functions of heat production in the model. Water loss in urine was driven by absorbed N left after being removed via milk. Model parameters were estimated simultaneously using observed fecal and urinary water output data from lactating Holstein cows (n = 670). The model was evaluated with data that were not used for model development and optimization (n = 377). The observations in both data sets were related to thermoneutral conditions. The model predicted drinking water intake, fecal, urinary, and total fresh manure water output with root mean square prediction errors as a percentage of average values of 18.1, 15.6, 30.6, and 14.6%, respectively. In all cases, >97% of the prediction error was due to random variability of data. The model can also be used to determine saliva production, heat and metabolic water production, respiratory cutaneous water losses, and size of major body water pools in lactating Holstein cows under thermoneutral conditions.     

Long-term effects of feeding monensin on methane production in lactating dairy cows

The objective of this study was to determine the long-term effects of feeding monensin on methane (CH₄) production in lactating dairy cows. Twenty-four lactating Holstein dairy cows (1.46 ± 0.17 parity; 620 ± 5.9 kg of live weight; 92.5 ± 2.62 d in milk) housed in a tie-stall facility were used in the study. The study was conducted as paired comparisons in a completely randomized design with repeated measurements in a color-coded, double-blind experiment. The cows were paired by parity and days in milk and allocated to 1 of 2 treatments: 1) the regular milking cow total mixed ration (TMR) with a forage-to-concentrate ratio of 60:40 (control TMR; placebo premix) vs. a medicated TMR (monensin TMR; regular TMR + 24 mg of Rumensin Premix/kg of dry matter) fed ad libitum. The animals were fed and milked twice daily (feeding at 0830 and 1300 h; milking at 0500 and 1500 h) and CH₄ production was measured prior to introducing the treatments and monthly thereafter for 6 mo using an open-circuit indirect calorimetry system. Monensin reduced CH₄ production by 7% (expressed as grams per day) and by 9% (expressed as grams per kilogram of body weight), which were sustained for 6 mo (mean, 458.7 vs. 428.7 ± 7.75 g/d and 0.738 vs. 0.675 ± 0.0141, control vs. monensin, respectively). Monensin reduced milk fat percentage by 9% (3.90 vs. 3.53 ± 0.098%, control vs. monensin, respectively) and reduced milk protein by 4% (3.37 vs. 3.23 ± 0.031%, control vs. monensin, respectively). Monensin did not affect the dry matter intake or milk yield of the cows. These results suggest that medicating a 60:40 forage-to-concentrate TMR with 24 mg of Rumensin Premix/kg of dry matter is a viable strategy for reducing CH₄ production in lactating Holstein dairy cows.     

Invited review: Antibiotic treatment of metritis in dairy cows: A systematic approach

Acute puerperal metritis (APM) is an acute systemic illness with fever ≥39.5°C and signs of toxemia due to an infection of the uterus occurring within 21 d after parturition. Because of the infectious nature of APM, antibiotics are considered beneficial for its treatment. Each use of an antimicrobial drug, however, is associated with selective pressure for the emergence of resistant bacteria. Hence, there is a significant need to encourage prudent use of antibiotics and alternative therapies to antibiotics. Therefore, the objective of this study was to systematically review the current literature on treatment of APM. A comprehensive and systematic literature search was conducted utilizing the PubMed and CAB Abstracts databases to identify literature focusing on the antibiotic therapy of puerperal metritis in the cow. After application of specific exclusion criteria, 21 publications comprising 23 trials remained for final evaluation. Data extraction revealed that the majority of the studies (n = 19) were attributable to the highest evidence level. Of 21 studies controlled, 11 had an untreated group and 3 a positive control group. The majority of the studies (n = 17) applied ceftiofur for the treatment of APM. Concerning the efficacy of ceftiofur, 7 studies observed clinical improvement, whereas none found improved reproductive performance. Fewer than half of the studies (n = 10) performed a bacteriological examination and only 4 implemented an antibiotic susceptibility test. Also, 3 studies (13.0%) described a self-cure rate per se. Little attention was given to the issue of bacterial resistance (n = 3), the need for reducing the application of antibiotics (n = 2), or guidelines for prudent use of antibiotics (n = 1). Our findings demonstrate that implementation of bacteriological examinations, sensitivity testing, and determination of minimum inhibitory concentrations, as well as reporting and discussion of critical issues (e.g., self-cure rates, resistance, prudent drug use), were suboptimal. On the other hand, the quality of studies on the treatment of APM was good, as indicated by evidence level 1. Nevertheless, more high-quality research considering self-cure rates is necessary to address critical issues related to APM and crucial to the dairy industry, such as resistance, prudent use of antibiotics, animal welfare, and cost-benefit ratios.     

Dietary cation-anion difference effects on performance and acid-base status of lactating dairy cows: a meta-analysis

A meta-analysis was conducted to examine potential empirical relationships between dietary cation-anion difference (DCAD) (Na + K - Cl) and the response of lactating dairy cows. The database was developed from 12 studies published between 1984 and 1997 that included a total of 17 trials, 69 dietary treatments, and 230 cows. Results indicated that DCAD affected performance of lactating dairy cows. Maximum milk yield and feed intake were reached when DCAD was 34 and 40 meq/100 g of feed dry matter, respectively. Blood pH and HCO3 concentrations increased with DCAD, indicating an improved acid-base balance of lactating dairy cows. Changes in urinary pH and urinary excretion of Na, K, and Cl were consistent with varying DCAD, thus dietary acidity or alkalinity. The effects of DCAD were likely mediated via modification of acid-base status in the cows.     

Feeding dairy cows bakery by-products enhanced nutrient digestibility,but affected fecal microbial composition and pH in a dose-dependent manner

We reported recently that adding bakery by-products (BP) to the diets of dairy cows up to 30% improved performance and rumen pH, but caused major shifts in the nutrient profile and availability, likely modifying nutrient degradation patterns throughout the gastrointestinal tract. The aim of this study was to investigate the effects of the gradual replacement of cereals by BP on the apparent total-tract digestibility (ATTD), the fermentation patterns, and the microbial community in feces of dairy cows. Twenty-four mid-lactating Simmental cows (149 ± 22.3 days in milk, 756 ± 89.6 kg of initial body weight) were fed a total mixed ration ad libitum (fresh feed was offered twice per day) containing a 50:50 ratio of forage to concentrate (dry matter basis) throughout the experiment. The trial lasted 5 wk, whereby the first week was used for baseline measurements, in which all cows received the same diet, without BP. Cows were then randomly allocated into 3 groups differing in the BP content of diets (0% BP, 15% BP, and 30% BP on a DM basis) and fed for 4 wk. Fecal samples were taken for analysis of pH, volatile fatty acids (VFA), and 16S rRNA gene sequencing. The inclusion of BP resulted in an increase of ether extract and sugars, and a reduction of starch and neutral detergent fiber in the diet. Feeding BP linearly increased the ATTD of almost all nutrients resulting in up to 2 kg more digestible organic matter intake (DOMI). Increasing BP level up to 30% increased fecal total VFA concentration and decreased the pH. The proportion of butyrate in feces increased linearly, but the proportion of all other VFA was not affected by BP-feeding. The richness and diversity indices of the fecal microbiota linearly declined by the inclusion of BP. The cellulolytic phyla Fibrobacteres decreased, whereas amylolytic phyla, such as Proteobacteria, increased. Overall, results showed that feeding BP linearly increased ATTD and DOMI, but impaired fecal microbial diversity and pH. In the interest of the optimization of BP inclusion in the dairy cows' feeding, a dietary level between 15 to 30% of BP might be a better compromise than 30% in terms of an enhanced DOMI and performance with still lowered risk of hindgut dysbiosis, but this will require further investigations.     

Short communication: the effects of histidine-supplemented drinking water on the performance of lactating dairy cows

An experiment was conducted to test the hypothesis that a sufficient proportion of histidine (His) included in the drinking water of lactating cows bypasses the rumen to have an effect on milk synthesis. Eight dairy cows (45 ± 15 d in milk) were given either 0 or 2.5 g/L of His in the drinking water in a crossover design of two 7-d periods. Cows were offered a corn and alfalfa silage-based total mixed ration for ad libitum intake. Water was provided ad libitum to each cow in an individual automatic drinking vessel with a flow meter attached. Water intake tended to increase from 85.1 to 92.1 L/d when His was added. Concentrations of His in plasma samples collected on the last day of each period tended to increase from 14.6 to 21.6 μM, corresponding to an estimated 0.4% bypass of the imbibed histidine. Other amino acid concentrations in plasma were not affected by His supplementation. Milk yield increased by 1.7 L/d with His treatment, lactose yield increased by 90 g/d, and there were tendencies for protein yield to increase, fat percentage to decrease, and protein to fat ratio to increase. An improvement in postruminal histidine flow can influence milk production and composition but the proportion of imbibed water that bypasses the rumen will have to be increased to take advantage of drinking water as a vehicle to transfer His postruminally.     

Prevalence of mastitis pathogens and their resistance against antimicrobial agents in dairy cows in Brandenburg, Germany

The primary objective of this study was to determine management practices concerning mastitis in Brandenburg, Germany, the prevalence of mastitis pathogens in dairy cows, and their resistance to selected antimicrobial agents. A further objective was to study the potential effect of parity and stage of lactation on the resistance of Staphylococcus aureus isolates against ampicillin. Milk samples for microbiological culture were collected from 4 groups of clinically healthy cows (first lactation, >1 lactation, >50 d in milk, and >250 d in milk; 8 cows/group) in 80 dairy herds. Resistance of gram-positive pathogens against 6 antimicrobial agents was tested using the broth microdilution method. Mastitis pathogens were isolated from 26.4% of the milk samples. Coagulase-negative staphylococci (CNS, 9.1% of quarters) and Corynebacterium bovis (7.3%) were the pathogens most frequently isolated. Among the major pathogens, Staph. aureus (5.7%) and Streptococcus uberis (1.0%) had the highest prevalence. Streptococcus agalactiae was isolated in samples from 29% of the herds. Although the prevalence of most pathogens was higher in older cows, the prevalence of CNS was higher in primiparous cows. Results of the mastitis control questionnaire showed that cows with clinical mastitis were transferred to a sick cow pen in 70% of the herds. Cephalosporins were the drug of first choice for treatment of clinical mastitis cases followed by fixed combinations of antimicrobial agents, β-lactamase-resistant penicillins, and penicillin. Most farmers treated cows 3 to 4 times per case. Cloxacillin, alone or in combination, and penicillin were most often used for dry-cow therapy. Antimicrobial resistance of the pathogens was within the range of other reports. Resistance of Staph. aureus to ampicillin increased significantly during the first lactation. Further research is required to determine the factors that lead to the selection of Staph. aureus strains that are resistant to ampicillin during the first lactation.     

Long-term effects of feeding monensin on milk fatty acid composition in lactating dairy cows

The objective of this study was to determine the long-term effects of feeding monensin on milk fatty acid (FA) profile in lactating dairy cows. Twenty-four lactating Holstein dairy cows (1.46 ± 0.17 parity; 620 ± 5.9 kg of live weight; 92.5 ± 2.62 d in milk) housed in a tie-stall facility were used in the study. The study was conducted as paired comparisons in a completely randomized block design with repeated measurements in a color-coded, double blind experiment. The cows were paired by parity and days in milk and allocated to 1 of 2 treatments: 1) the regular milking cow total mixed ration (TMR) with a forage-to-concentrate ratio of 60:40 (control TMR; placebo premix) vs. a medicated TMR [monensin TMR; regular TMR + 24 mg of Rumensin Premix per kg of dry matter (DM)] fed ad libitum. The animals were fed and milked twice daily (feeding at 0830 and 1300 h; milking at 0500 and 1500 h). Milk samples were collected before the introduction of treatments and monthly thereafter for 6 mo and analyzed for FA composition. Monensin reduced the percentage of the short-and medium-chain saturated FA 7:0, 9:0, 15:0, and 16:0 in milk fat by 26, 35, 19, and 6%, respectively, compared with the control group. Monensin increased the percentage of the long-chain saturated FA in milk fat by 9%, total monounsaturated FA by 5%, total n-6 polyunsaturated FA (PUFA) by 19%, total n-3 PUFA by 16%, total cis-18:1 by 7%, and total conjugated linoleic acid (CLA) by 43% compared with the control group. Monensin increased the percentage of docosahexaenoic acid (22:6n-3), docosapentaenoic acid (22:5n-3), and cis-9, trans-11 CLA in milk fat by 19, 13, and 43%, respectively, compared with the control. These results suggest that monensin was at least partly effective in inhibiting the biohydrogenation of unsaturated FA in the rumen and consequently increased the percentage of n-6 and n-3 PUFA and CLA in milk, thus enhancing the nutritional properties of milk with regard to human health.     

Effect of milking frequency on the behavior and productivity of lactating dairy cows

The objective of this study was to determine the effect of milking frequency on the behavioral patterns and productivity of lactating dairy cows. Twelve freestall-housed, lactating Holstein dairy cows (7 primiparous and 5 multiparous) were exposed to each of 2 treatments (over 21-d periods) in a replicated crossover design. Treatments were milking frequency of 2×/d (at 0600 and 1800 h) or 3×/d (at 0600, 1400, and 2200 h). Milk production, feeding, lying, and rumination behavior were monitored for each animal for the last 7 d of each treatment period. Milk samples were collected for the last 3 d of each period for milk component analysis. The results indicated that cows milked 3×/d produced 2.9 kg/d more milk than those milked 2×/d. Primiparous cows consumed 3.9 kg/d less dry matter (DM) than did multiparous cows. The extra time (14.6 min/d) required for milking 3×/d altered the distribution of cow behavioral activity throughout the day. Although this did not affect total daily lying or rumination time, we observed a tendency for cows milked 2×/d to spend less time (224.6 vs. 237.5 min/d) feeding and, thus, those cows tended to consume their feed at a faster rate (0.13 vs. 0.12 kg of DM/min) than cows milked 3×/d. For multiparous cows, the increase in feeding activity was facilitated through having longer (40.1 vs. 36.8 min/meal) and numerically larger meals (4.8 vs. 4.6 kg of DM/meal) when milked 3×/d. Alternatively, primiparous cows consumed smaller (2.9 vs. 3.2 kg of DM/meal) and more frequent meals (9.1 vs. 7.7 meals/d) throughout the day when milked 3×/d, resulting in a tendency for greater DM intake (24.7 vs. 23.6 kg/d) compared with primiparous cows milked 2×/d. These results indicate that under 3×/d milking schedules, primiparous cows will positively adjust their feeding behavior to achieve similar production increases as multiparous cows. In summary, milking 3×/d can be used to improve production; however, greater milking frequency elicits varying effects on the behavior of primiparous and multiparous cows, suggesting that grouping and management of cows based on parity may be beneficial.     

Symposium review: The uterine microbiome associated with the development of uterine disease in dairy cows

Until 2010, our knowledge of the uterine microbiome in cows that developed uterine disease relied almost exclusively on culture-dependent studies and mostly included cows with clinical endometritis (i.e., with purulent uterine discharge). Those studies consistently found a strong positive correlation between Trueperella pyogenes and clinical endometritis, whereas other pathogens such as Escherichia coli, Fusobacterium necrophorum, Prevotella melaninogenica, and Bacteroides spp. were also commonly cocultured. In contrast, Streptococcus spp., Staphylococcus spp., and Bacillus spp. were usually isolated from healthy cows. Starting in 2010, culture-independent studies using PCR explored the microbiome of cows with metritis and clinical endometritis, and observed that E. coli was a pioneer pathogen that predisposed cows to infection with F. necrophorum, which was strongly associated with metritis, and to infection with T. pyogenes, which was strongly associated with clinical endometritis. Starting in 2011, culture-independent studies using metagenomic sequencing expanded our knowledge of the uterine microbiome. It has been shown that cows have bacteria in the uterus even before calving, they have an established uterine microbiome within 20 min of calving, and that the microbiome structure is identical between cows that develop metritis and healthy cows until 2 d postpartum, after which the bacterial structure of cows that developed metritis deviates in favor of greater relative abundance of Bacteroidetes and Fusobacteria and lesser relative abundance of Proteobacteria and Tenericutes. The shift in the uterine microbiome in cows that develop metritis is characterized by a loss of heterogeneity and a decrease in bacterial richness. At the genus level, Bacteroides, Porphyromonas, and Fusobacterium have the strongest association with metritis. At the species level, we observed that Bacteroides pyogenes, Porphyromonas levii, and Helcococcus ovis were potential emerging uterine pathogens. Finally, we have shown that the hematogenous route is a viable route of uterine infection with uterine pathogens. Herein, we propose that metritis is associated with a dysbiosis of the uterine microbiota characterized by decreased richness, and an increase in Bacteroidetes and Fusobacteria, particularly Bacteroides, Porphyromonas, and Fusobacterium.     

Rumen fermentation and production effects of Origanum vulgare L. leaves in lactating dairy cows

A lactating cow trial was conducted to study the effects of dietary addition of oregano leaf material (Origanum vulgare L.; OV; 0, control vs. 500 g/d) on ruminal fermentation, methane production, total tract digestibility, manure gas emissions, N metabolism, organoleptic characteristics of milk, and dairy cow performance. Eight primiparous and multiparous Holstein cows (6 of which were ruminally cannulated) were used in a crossover design trial with two 21-d periods. Cows were fed once daily. The OV material was top-dressed and mixed with a portion of the total mixed ration. Cows averaged 80 ± 12.5 d in milk at the beginning of the trial. Rumen pH, concentration of total and individual volatile fatty acids, microbial protein outflow, and microbial profiles were not affected by treatment. Ruminal ammonia-N concentration was increased by OV compared with the control (5.3 vs. 4.3 mM). Rumen methane production, which was measured only within 8 h after feeding, was decreased by OV. Intake of dry matter (average of 26.6 ± 0.83 kg/d) and apparent total tract digestibly of nutrients did not differ between treatments. Average milk yield, milk protein, lactose, and milk urea nitrogen concentrations were unaffected by treatment. Milk fat content was increased and 3.5% fat-corrected milk yield tended to be increased by OV, compared with the control (3.29 vs. 3.12% and 42.4 vs. 41.0 kg/d, respectively). Fat-corrected (3.5%) milk feed efficiency and milk net energy for lactation (NE_L) efficiency (milk NE_L ÷ NE_L intake) were increased by OV compared with the control (1.64 vs. 1.54 kg/kg and 68.0 vs. 64.4%, respectively). Milk sensory parameters were not affected by treatment. Urinary and fecal N losses, and manure ammonia and methane emissions were unaffected by treatment. Under the current experimental conditions, supplementation of dairy cow diets with 500 g/d of OV increased milk fat concentration, feed and milk NE_L efficiencies, and tended to increase 3.5% fat-corrected milk yield. The sizable decrease in rumen methane production with the OV supplementation occurred within 8 h after feeding and has to be interpreted with caution due to the large within- and between-animal variability in methane emission estimates. The OV was introduced into the rumen as a pulse dose at the time of feeding, thus most likely having larger effect on methane production during the period when methane data were collected. It is unlikely that methane production will be affected to the same extent throughout the entire feeding cycle.     

Effects of a combination of feed additives on methane production, diet digestibility, and animal performance in lactating dairy cows

Two experiments were conducted to assess the effects of a mixture of dietary additives on enteric methane production, rumen fermentation, diet digestibility, energy balance, and animal performance in lactating dairy cows. Identical diets were fed in both experiments. The mixture of feed additives investigated contained lauric acid, myristic acid, linseed oil, and calcium fumarate. These additives were included at 0.4, 1.2, 1.5, and 0.7% of dietary dry matter, respectively (treatment ADD). Experimental fat sources were exchanged for a rumen inert source of fat in the control diet (treatment CON) to maintain isolipidic rations. Cows (experiment 1, n = 20; experiment 2, n = 12) were fed restricted amounts of feed to avoid confounding effects of dry matter intake on methane production. In experiment 1, methane production and energy balance were studied using open-circuit indirect calorimetry. In experiment 2, 10 rumen-fistulated animals were used to measure rumen fermentation characteristics. In both experiments animal performance was monitored. The inclusion of dietary additives decreased methane emissions (g/d) by 10%. Milk yield and milk fat content tended to be lower for ADD in experiment 1. In experiment 2, milk production was not affected by ADD, but milk fat content was lower. Fat- and protein-corrected milk was lower for ADD in both experiments. Milk urea nitrogen content was lowered by ADD in experiment 1 and tended to be lower in experiment 2. Apparent total tract digestibility of fat, but not that of starch or neutral detergent fiber, was higher for ADD. Energy retention did not differ between treatments. The decrease in methane production (g/d) was not evident when methane emission was expressed per kilogram of milk produced. Feeding ADD resulted in increases of C12:0 and C14:0 and the intermediates of linseed oil biohydrogenation in milk in both experiments. In experiment 2, ADD-fed cows tended to have a decreased number of protozoa in rumen fluid when compared with that in control cows. Total volatile fatty acid concentrations were lower for ADD, whereas molar proportions of propionate increased at the expense of acetate and butyrate.     

Susceptibility of Escherichia coli isolated from uteri of postpartum dairy cows to antibiotic and environmental bacteriophages. Part I: Isolation and lytic activity estimation of bacteriophages

The objective of this study was to isolate bacteriophages from environmental samples of 2 large commercial dairy farms using Escherichia coli isolated from the uteri of postpartum Holstein dairy cows as hosts. A total of 11 bacteriophage preparations were isolated from manure systems of commercial dairy farms and characterized for in vitro antimicrobial activity. In addition, a total of 57 E. coli uterine isolates from 5 dairy cows were phylogenetically grouped by triplex PCR. Each E. coli bacterial host from the uterus was inoculated with their respective bacteriophage preparation at several different multiplicities of infections (MOI) to determine minimum inhibitory MOI. The effect of a single dose (MOI = 10²) of bacteriophage on the growth curve of all 57 E. coli isolates was assessed using a microplate technique. Furthermore, genetic diversity within and between the different bacteriophage preparations was assessed by bacteriophage purification followed by DNA extraction, restriction, and agarose gel electrophoresis. Phylogenetic grouping based on triplex PCR showed that all isolates of E. coli belonged to phylogroup B1. Bacterial growth was completely inhibited at considerably low MOI, and the effect of a single dose (MOI = 10²) of bacteriophage preparations on the growth curve of all 57 E. coli isolates showed that all bacteriophage preparations significantly decreased the growth rate of the isolates. Bacteriophage preparation 1230-10 had the greatest antimicrobial activity and completely inhibited the growth of 71.7% (n = 57) of the isolates. The combined action of bacteriophage preparations 1230-10, 6375-10, 2540-4, and 6547-2, each at MOI = 10², had the broadest spectrum of action and completely inhibited the growth (final optical density at 600 nm ≤0.1) of 80% of the E. coli isolates and considerably inhibited the growth (final optical density at 600 nm ≤0.2) of 90% of the E. coli isolates. Restriction profile analysis demonstrated that all 4 phage preparations contained bacteriophages that were genetically distinct from each other according to the banding pattern of the fragments. The combination of several different bacteriophages can improve the spectrum of action, and the results of this study suggest that bacteriophages 1230-10, 6375-10, 2540-4, and 6547-2 should be used in combination as a cocktail.     

The effect of internal teat sealant products (Teatseal and Orbeseal) on intramammary infection,clinical mastitis,and somatic cell counts in lactating dairy cows: A meta-analysis

The objective of this study was to assess the efficacy of internal teat sealant products containing bismuth subnitrate (Teatseal and Orbeseal; Pfizer Animal Health, West Ryde, Australia) when used alone, or in the presence of antibiotic dry cow therapy (ADCT), before or at drying off on the incidence of new intramammary infections (IMI), clinical mastitis, and milk somatic cell count (SCC) during lactation. The literature search identified 18 English-language publications on the use of Teatseal in dairy cattle. A total of 12 studies with 17 subtrials or comparisons including 13 positive control subtrials (internal teat sealant and ADCT vs. ADCT) and 4 negative control subtrials (internal teat sealant vs. untreated) examining IMI were included in the analysis. Internal teat sealants, alone or in the presence of ADCT, reduced the risk of acquiring new IMI after calving by 25% [risk ratio (RR) = 0.75; 95% confidence interval (CI): 0.67 to 0.83]. Internal teat sealants reduced the risk of IMI by 73% compared with untreated cows (RR = 0.27; 95% CI: 0.13 to 0.55). The results of both meta-analyses of IMI, with positive and negative controls, were heterogeneous [I² (a statistic that describes the proportion of total variation in study effect estimates that is due to heterogeneity) = 65.4 and 92.1%]. No farm or cow factors studied significantly contributed to the heterogeneity of the results. A total of 16 studies (21 subtrials), including 14 positive control subtrials and 7 negative control subtrials, examining clinical mastitis were included in the analysis. Internal teat sealants alone and in the presence of ADCT reduced the risk of clinical mastitis after calving in lactating cows by 29% (RR = 0.71; 95% CI: 0.62 to 0.82), and 48% (RR = 0.52; 95% CI: 0.37 to 0.75), respectively. The results of the meta-analysis on clinical mastitis with positive controls were homogeneous (I² = 33.6%), whereas the results of studies with negative controls were heterogeneous (I² = 60.4%). No farm or cow factors studied that had sufficient data to evaluate significantly contributed to the heterogeneity of the results. The estimated linear score (LS) of milk SCC after calving in published studies (n = 3) and for studies that provided raw data (n = 2), was significantly lower for cattle treated with internal teat sealants and ADCT in 3 studies than for cattle treated with internal teat sealants only. The estimated LS of pooled raw data of 3 studies from 32 herds showed that the LS of cows treated with internal teat sealant and ADCT was not significantly different than those treated with ADCT only. This study found that the application of internal teat sealants in the presence of ADCT or the use of internal teat sealants alone at dry off significantly reduced the incidence of IMI and clinical mastitis in lactating dairy cows compared with respective control groups. Further studies are needed to investigate the effect of internal teat sealants on postpartum milk SCC in lactating dairy cows.     

The effects of varying gossypol intake from whole cottonseed and cottonseed meal on lactation and blood parameters in lactating dairy cows

Effects of varying amounts of gossypol from whole Upland cottonseed (WCS) and cottonseed meal (CSM) were evaluated in 40 midlactation Holstein cows. After 14 d of pretreatment, cows were assigned to 1 of the 5 treatments for 84 d: control (no gossypol), 931 mg/kg total gossypol (TG) and 850 mg/kg free gossypol (FG) from WCS (moderate TG and high FG); 924 mg/kg TG and 91 mg/kg FG from CSM (moderate TG and low FG), 945 mg/kg TG and 479 mg/kg FG with equal amounts of TG from WCS and CSM (moderate TG and FG), or 1894 mg/kg TG and 960 mg/kg FG with equal amounts of TG from WCS and CSM (high TG and FG). Concentrations of plasma gossypol (PG) and its isomers were directly proportional to FG intake. Concentrations of PG reached a plateau after 28 d on treatment, and they were highest in cows receiving a diet with high TG and FG. Erythrocyte fragility differed among treatments and increased with increasing FG intake. Plasma gossypol returned to negligible concentrations 28 d after withdrawal of cottonseed products from the high TG and FG diet. Serum vitamin A was similar among treatments, but vitamin E increased with increasing FG intake. Serum enzymes were generally unaffected by treatments, but urea N increased in diets higher in TG and FG. Intake of dry matter was higher for the diet high in TG and FG than for the control diet, but was similar for other treatments. Cows receiving the high TG and FG diet produced more milk and 3.5% fat-corrected milk, with no changes in milk composition. Feeding a diet containing 1894 mg/kg TG and 960 mg/kg FG for 84 d increased PG concentrations and erythrocyte fragility and resulted in minor changes in blood metabolites and enzymes, but no detrimental effect on lactation performance was observed. Indicators of liver, kidney, and muscle cell viability suggest that the higher amounts of gossypol consumed in this study had only minor effects on those tissues in lactating dairy cows.     

Type of cottonseed and level of gossypol in diets of lactating dairy cows: effects on lactation performance and plasma gossypol

Objectives were to determine the effects of feeding whole linted Upland (WUP) and cracked Pima cottonseed (BUPCP) to lactating cows on plasma gossypol concentrations and lactation performance. Holstein cows (n = 813) from three commercial dairies were assigned to one of two diets starting at 13+/-11 d in milk (DIM) for a 170-d experimental period. Cottonseed was included at 10% of the diet dry matter, and treatments consisted of replacing WUP with a 1:2 blend of BUPCP. Blood was collected from all cows at 61 and 91 DIM and from a subset of 56 cows on one dairy at 10, 33, 61, 91, 120, and 152 DIM. Intakes of free gossypol increased 32% in cows receiving the BUPCP diet, and concentrations of total gossypol (TG), as well as the proportion of TG as minus (-) isomer in plasma, were higher for cows fed BUPCP than WUP. Plasma TG increased linearly with increasing DIM, but cows fed the BUPCP, especially multiparous cows, had a higher increase in plasma TG over time. Yields of milk and milk components did not differ between the two treatments, although, yields of milk and milk components were greater in cows with higher plasma TG. Replacement of WUP cottonseed with BUPCP cottonseed increased plasma gossypol, but dry matter intake and yields of milk and milk components were not affected.     

Discrepancies among healthy,subclinical mastitic,and clinical mastitic cows in fecal microbiome and metabolome and serum metabolome

Mastitis is generally considered a local inflammatory disease caused by the invasion of exogenous pathogens and resulting in the dysbiosis of microbiota and metabolites in milk. However, the entero-mammary pathway theory may establish a possible link between some endogenous gut bacteria and the occurrence and development of mastitis. In the current study, we attempted to investigate differences in the gut microbiota profile and metabolite composition in gut and serum from healthy cows and those with subclinical mastitis and clinical mastitis. Compared with those of healthy cows, the microbial community diversities in the feces of cows with subclinical mastitis (SM) and clinical mastitis (CM) were lower. Lower abundance of Bifidobacterium, Romboutsia, Lachnospiraceae_NK3A20_group, Coprococcus, Prevotellaceae_UCG-003, Ruminococcus, and Alistipes, and higher abundance of the phylum Proteobacteria and the genera Escherichia-Shigella and Streptococcus were observed in CM cows. Klebsiella and Paeniclostridium were significantly enriched in the feces of SM cows. Several similarities were observed in feces and serum metabolites in mastitic cows. Higher levels of proinflammatory lipid products (20-trihydroxy-leukotriene-B4, 13,14-dihydro-15-keto-PGE₂, and 9,10-dihydroxylinoleic acids) and lower levels of metabolites involved in secondary bile acids (deoxycholic acid, 12-ketolithocholic acid), energy (citric acid and 3-hydroxyisovalerylcarnitine), and purine metabolism (uric acid and inosine) were identified in both SM and CM cows. In addition, elevated concentrations of IL-1β, IL-6, tumor necrosis factor-α and decreased concentrations of glutathione peroxidase and superoxide dismutase were detected in the serum of SM and CM cows. Higher serum concentrations of triglyceride and total cholesterol and lower concentrations of high-density lipoproteins in mastitic cows might be related to changes in the gut microbiota and metabolites. These findings suggested a significant difference in the profile of feces microbiota and metabolites in cows with different udder health status, which might increase our understanding of bovine mastitis.     

Selenium supplementation of lactating dairy cows: effect on selenium concentration in blood, milk, urine, and feces

The objectives were to determine effects of graded levels of selenized yeast derived from a specific strain of Saccharomyces cerevisiae (CNCM I-3060) on animal performance and in selenium concentrations in the blood, milk, feces, and urine of dairy cows compared with sodium selenite; and to provide preliminary data on the proportion of selenium as selenomethionine in the milk and blood. Twenty Holstein cows were used in a 5 × 5 Latin square design study in which all cows received the same total mixed rations, which varied only in source or concentration of dietary selenium. There were 5 experimental treatments. Total dietary selenium of treatment 1, which received no added selenium, was 0.15 mg/kg of dry matter, whereas values for treatments 2, 3, and 4, derived from selenized yeast, were 0.27, 0.33, and 0.40 mg/kg of dry matter, respectively. Treatment 5 contained 0.25 mg of selenium obtained from sodium selenite/kg of dry matter. There were no significant treatment effects on animal performance, and blood chemistry and hematology showed few treatment effects. Regression analysis noted significant positive linear effects of increasing dietary selenium derived from selenized yeast on selenium concentrations in the milk, blood, urine, and feces. In addition, milk selenium results indicated improved bioavailability of selenium from selenized yeast, compared with sodium selenite. Preliminary analyses showed that compared with sodium selenite, the use of selenized yeast increased the concentration of selenomethionine in the milk and blood. There was no indication of adverse effects on cow health associated with the use of selenized yeast.