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The objective for this study was to evaluate the supplementation with different non-fibrous carbohydrate (NFC) sources for grazing dairy cows in diets with high or low concentrations of rumen undegradable protein (RUP). Twelve multiparous cows averaging 30kg/d of milk production and 45±23 days in milk in a 4x4 Latin square design with 2 × 2 factorial arrangement were used. The difference in NFC content of the diets was achieved using ground corn (GC) or dried citrus pulp (DCP). Dietary RUP was altered by the partial replacement of soybean meal (LRUP) by roasted whole soybean and corn gluten meal (HRUP). Greater dry matter intake (DMI) was observed in DCP treatments compared to GC treatments (19.2 and 17.7kg/day of DM, respectively). Milk fat concentration was lower for LRUP treatments (3.4%) when compared to HRUP (3.6%). The milk protein concentration was greater for GC treatments (2.9%) in relation to DCP treatments (2.8%). Lower milk ureic nitrogen was observed when DCP diets were fed compared to those with GC (17.8 and 21.2mg/dL, respectively). The use of DCP as CNF source increased dry matter intake as well as milk fat concentration, although no differences in milk production were observed for RUP level.

Publisher Copyright: © 2021 American Dairy Science Association The objective of this experiment was to investigate the effect of dietary levels of digestible histidine (dHis) and MP on lactational performance and plasma and muscle concentrations of free AA in dairy cows. A randomized block design experiment was conducted with 48 Holstein cows, including 20 primiparous, averaging (±SD) 103 ± 22 d in milk and 45 ± 9 kg/d milk yield at the beginning of the experiment. A 2-wk covariate period preceded 12 experimental wk, of which 10 wk were for data and sample collection. Experimental treatments were (1) MP-adequate (MPA) diet with 2.1% dHis of MP (MPA2.1), (2) MPA with 3.0% dHis (MPA3.0), (3) MP-deficient (MPD) diet with 2.1% dHis (MPD2.1), and (4) MPD with 3.0% dHis (MPD3.0). Actual dHis supply was estimated at 64, 97, 57, and 88 g/d, respectively. Diets supplied MP at 110% (MPA) and 96% (MPD) of NRC 2001 dairy model requirements calculated based on DMI and production data during the experiment. Dry matter intake and milk yield data were collected daily, milk samples for composition and blood samples for AA analysis were collected every other week, and muscle biopsies at the end of covariate period, and during wk 12 of the experiment. The overall DMI was not affected by dHis or MP level. Milk yield tended to be increased by 3.0% dHis compared with 2.1% dHis. Milk true protein concentration and yield were not affected by treatments, whereas milk urea nitrogen concentration was lower for MPD versus the MPA diet. Milk fat concentration was lower for MPD versus MPA. There was a MP × dHis interaction for milk fat yield and energy-corrected milk; milk fat was lower for MPD3.0 versus MPD2.1, but similar for cows fed the MPA diet regardless of dHis level whereas energy-corrected milk was greater for MPA3.0 versus MPA2.1 but tended to be lower for MPD3.0 versus MPD2.1. Plasma His concentration was greater for cows fed dHis3.0, and concentration of sum of essential AA was greater, whereas carnosine, 1-Methyl-His and 3-Methyl-His concentrations were lower for cows fed MPA versus MPD diet. Muscle concentration of His was greater for cows fed dHis3.0 treatment. The apparent efficiency of His utilization was increased at lower MP and His levels. Overall, cows fed a corn silage-based diet supplying MP at 110% of NRC (2001) requirements tended to have increased ECM yield and similar milk protein yield to cows fed a diet supplying MP at 96% of requirements. Supplying dHis at 3.0% of MP (or 86 and 96 g/d, for MPD3.0 and MPA3.0, respectively) tended to increase milk yield and increased plasma and muscle concentrations of His but had minor or no effects on other production variables in dairy cows.

The objective of this study was to investigate the effects of supplementation of an exogenous enzyme preparation (EEP) on performance, total-tract digestibility of nutrients, plasma AA profile, and milk fatty acids composition in lactating dairy cows fed a reduced-starch diet compared with a normal-starch diet (i.e., positive control). Forty-eight Holstein cows (28 primiparous and 20 multiparous) were enrolled in a 10-wk randomized complete block design experiment with 16 cows per treatment. Treatments were as follows: (1) normal-starch diet (control) containing (% dry matter basis) 24.8% starch and 33.0% neutral detergent fiber (NDF), (2) reduced-starch diet (RSD) containing 18.4% starch and 39.1% NDF, or (3) RSD supplemented with 10 g/cow per day of an EEP (ENZ). The EEP contained amylolytic and fibrolytic activities and was top-dressed on the total mixed ration at the time of feeding. Compared with normal-starch diet, dry matter intake and milk and energy-corrected milk (ECM) yields were lower (on average by 7.1, 9.5, and 7.2%, respectively) for cows on the RSD treatments. Concentrations, but not yields, of milk fat and total solids were increased by RSD. Energy-corrected milk feed efficiency did not differ among treatments. Total-tract digestibility of NDF tended to increase by RSD treatments. Plasma AA concentrations were not affected by treatment, except that of 3-methylhistidine was increased by ENZ, compared with RSD. Blood glucose concentration tended to be lower in cows on the RSD treatments, but ENZ increased glucose and tended to increase insulin concentrations at 4 h after feeding when compared with RSD. Cows on the RSD treatments had decreased concentrations of de novo fatty acids and tended to have increased concentrations of preformed fatty acids in milk. Overall, decreasing dietary starch concentration by 26% decreased dry matter intake, milk, and ECM yields, but ECM feed efficiency was not different among treatments. The negative effects of reducing dietary starch on production were not attenuated by the EEP.

Microbial syntrophy (obligate metabolic mutualism) is the hallmark of energy-constrained anaerobic microbial ecosystems. For example, methanogenic archaea and fermenting bacteria coexist by interspecies hydrogen transfer in the complex microbial ecosystem in the foregut of ruminants; however, these synergistic interactions between different microbes in the rumen are seldom investigated. We hypothesized that certain bacteria and archaea interact and form specific microbial cohorts in the rumen. To this end, we examined the total (DNA-based) and potentially metabolically active (cDNA-based) bacterial and archaeal communities in rumen samples of dairy cows collected at different times in a 24 h period. Notably, we found the presence of distinct bacterial and archaeal networks showing potential metabolic interactions that were correlated with molar proportions of specific volatile fatty acids (VFAs). We employed hypothesis-driven structural equation modeling to test the significance of and to quantify the extent of these relationships between bacteria-archaea-VFAs in the rumen. Furthermore, we demonstrated that these distinct microbial networks were host-specific and differed between cows indicating a natural variation in specific microbial networks in the rumen of dairy cows. This study provides new insights on potential microbial metabolic interactions in anoxic environments that have broader applications in methane mitigation, energy conservation, and agricultural production.

This study's aim is to evaluate the substitution of soybean meal with a yeast-derived protein (YDP) in the diet of F1 Holstein (Bos Taurus) x Gyr (Bos indicus) dairy cows under rotational grazing (Guinea Grass, Panicum maximum cv. Mombaca) and to analyze the effects on milk yield, milk solids percentage, blood parameters, and feed efficiency (FE). Sixteen Holstein X Gyr (F1) multiparous cows with an average of 30 ± 1.8 kg/d (Mean ± SD) of milk, 489 ± 22 kg of body weight, and 62 ± 26 days in milk (DIM), at the beginning of the study, were assigned to four squares based on their DIM, milk yield, and parity. The experimental diets were based on the substitution of soybean meal with a YDP in equal proportions: treatment YDP0 (without a YDP), treatment YDP150 (substitution of 150 g of soybean meal with a YDP), treatment YDP300 (substitution of 300 g of soybean meal with a YDP), and treatment YDP450 (substitution of 450 g of soybean meal with a YDP). The substitution of soybean meal with a YDP in the diets of dairy cows under rotational grazing linearly decreased milk and solids production and affected the neutral detergent fiber (NDF) intake, without affecting the total feed intake. No difference was found in the microbial protein synthesis between treatments. FE tended to be lower at high inclusions of the YDP.

The aim of this study was to evaluate the frequency of isolation of agents causing subclinical mastitis in a herd and to estimate production losses associated with SCC> 200,000cells/mL. Three 7-day interval microbiological cultures were performed in all lactating animals from the same farm that was evaluated from June to July. To evaluate the negative and positive isolation frequencies between the lactation phases, a Chi-square test was performed. Simple linear regressions were performed to evaluate the lactation curve of animals grouped by pathogens isolated from negative cows in the microbiological culture and with SCC 200.000 cél./mL. Três cultivos microbiológicos intervalados por sete dias foram realizados em todos os animais em lactação da propriedade avaliada, no período de junho a julho. Para avaliar as frequências de isolamento negativo e positivo entre as fases da lactação, foi realizado um teste de qui-quadrado. Foram realizadas regressões lineares simples para avaliar a curva de lactação dos animais agrupados por patógenos isolados em relação a vacas negativas na cultura microbiológica e com CCS < 200.000 cél./mL. Os coeficientes das regressões foram utilizados para estimar as perdas de produção entre os grupos. Vacas com resultado negativo na microbiologia, mas com CCS ≥ 200.000 cél./mL, positivas na microbiologia com CCS < 200.000 cél./mL e positivas com CCS ≥ 200.000 cél./mL, produziram por dia, respectivamente, -3,5; -0,5 e -4,27kg de leite em relação às vacas negativas com CCS < 200.000 cél./mL. Vacas infectadas com células leveduriformes, Staphylococcus coagulase negativa, Staphylococcus aureus e Streptococcus ambientais produziram, respectivamente, -3,42; -0,5; -0,168 e -2,5kg de leite, comparadas a vacas negativas com CCS < 200.000 cél./mL. A CCS, indicativa de reação inflamatória, encontra-se diretamente associada às perdas de produção de leite, assim como a presença do microrganismo na glândula mamária.

Rumination time is a relevant marker for health evaluation. The use of rumination monitoring systems can decrease labor costs when measuring rumination time. The objectives of this study were to: i) validate the Hr-Tag rumination monitoring system for estimating rumination time in Zebu calves during the weaning period and ii) to evaluate the Hr-Tag capacity to identify differences in daily rumination patterns during the weaning period. Thirty-two Gyr calves averaging 69 ± 7.6 kg body weight and 77.0 ± 4.0 days were used. Calves were fed 4.7 ± 0.5 L/day of whole milk and ad libitum commercial textured starter ration up to 83 days old (pre-weaning period). During the weaning period (84–86 days old) the same starter was used, but corn silage was also available and only half the milk dose was offered. After weaning (87–92 days old, post-weaning period) only corn silage and starter were used. For the Hr-Tag system validation, the rumination time obtained by Hr-Tag was compared with visual observations at four 2 h intervals (0600 to 0800, 0800 to 1000, 1400 to 1600, and 1600 to 1800 h) at pre- and post-weaning. Two observers were used in all visual evaluations. To compare daily rumination pattern at pre-weaning, weaning and post-weaning, only the Hr-Tag data was used. Rumination time values between observers were highly correlated (ρ = 0.956; P

The aim of this study was to identify possible effects of different vaccination strategies (concomitantly or not) against brucellosis and clostridia on intake, performance, feeding behavior, blood parameters, and immune responses of dairy heifers calves. Fifty heifers calves were enrolled [38 Gyr (Zebu, Bos taurus indicus) and 12 5/8 Holstein × Gyr]. At 120 d of age, animals were randomly distributed among 3 groups: B (n = 18), vaccinated against brucellosis; C (n = 14), vaccinated against clostridia and CB (n = 18), vaccinated concomitantly for both. Rectal and thermographic temperatures were evaluated on days −1, 0, 1, 2, 3, 5, 7,10, 14, and 28 relatives to the vaccination day. Feed and water intake, body weight (BW), and feeding behavior were monitored daily by an electronic feeding system. Blood was sampled on days 0, 3, 7, 14, and 28, relative to the vaccination day for determination of glucose and β -hydroxybutyrate (BHBA) concentrations. Blood sampled on day 0 (prevaccination) and on days 28 and 42 were used to evaluate the immune response against Brucella abortus and clostridia. There was an increase in rectal temperature between the first and the third day postvaccination in the 3 groups. The thermography revealed an increase of local temperature for 7 d on groups B and CB. Group C had increased local temperature for a longer period, lasting for up to 14 d. Dry mater intake was reduced for groups B and CB, but no alteration was observed for group C. No alterations regarding initial BW, final BW, average daily weight gain, and feed efficiency were observed. No differences were observed for the 3 vaccination groups for blood parameters throughout the evaluation period. The concomitant vaccination against brucellosis and clostridia led to lower neutralizing antibody titers against epsilon toxin of Clostridium perfringens and botulinum toxin type C of C. botulinum (C > CB > B). When cellular proliferation assay and serological tests to B. abortus were evaluated, no differences were observed between groups B and CB. The present results indicate that the concomitant vaccination against brucellosis and clostridia has no relevant impact on the intake, performance, and feeding behavior of dairy calves. However, the concomitant vaccination of vaccines against these 2 pathogens impacts animal immunity against clostridial infections.

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 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 kp 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.

This study investigated the effects of an amylase-enabled corn silage on lactational performance, enteric CH4 emission, and rumen fermentation of lactating dairy cows. Following a 2-wk covariate period, 48 Holstein cows were blocked based on parity, days in milk, milk yield (MY), and CH4 emission. Cows were randomly assigned to 1 of 2 treatments in an 8-wk randomized complete block design experiment: (1) control corn silage (CON) from an isogenic corn without α-amylase trait and (2) Enogen hybrid corn (Syngenta Seeds LLC) harvested as silage (ECS) containing a bacterial transgene expressing α-amylase (i.e., amylase-enabled) in the endosperm of the grain. The ECS and CON silages were included at 40% of the dietary dry matter (DM) and contained, on average, 43.3 and 41.8% DM and (% DM) 36.7 and 37.5% neutral detergent fiber, and 36.1 and 33.1% starch, respectively. Rumen samples were collected from a subset of 10 cows using the ororuminal sampling technique on wk 3 of the experimental period. Enteric CH4 emission was measured using the GreenFeed system (C-Lock Inc.). Dry matter intake (DMI) was similar between treatments. Compared with CON, MY (38.8 vs. 40.8 kg/d), feed efficiency (1.47 vs. 1.55 kg of MY/kg of DMI), and milk true protein (1.20 vs. 1.25 kg/d) and lactose yields (1.89 vs. 2.00 kg/d) were increased, whereas milk urea nitrogen (14.0 vs. 12.7 mg/dL) was decreased, with the ECS diet. No effect of treatment on energy-corrected MY (ECM) was observed, but a trend was detected for increased ECM feed efficiency (1.45 vs. 1.50 kg of ECM/kg of DMI) for cows fed ECS compared with CON-fed cows. Daily CH4 emission was not affected by treatment, but emission intensity was decreased with the ECS diet (11.1 vs. 10.3 g/kg of milk, CON and ECS, respectively); CH4 emission intensity on ECM basis was not different between treatments. Rumen fermentation, apart from a reduced molar proportion of butyrate in ECS-fed cows, was not affected by treatment. Apparent total-tract digestibility of nutrients and urinary and fecal nitrogen excretions, apart from a trend for increased DM digestibility by ECS-fed cows, were not affected by treatment. Overall, ECS inclusion at 40% of dietary DM increased milk, milk protein, and lactose yields and feed efficiency, and tended to increase ECM feed efficiency but had no effect on ECM yield in dairy cows. The increased MY with ECS led to a decrease in enteric CH4 emission intensity, compared with the control silage.

Asparagopsis taxiformis (AT) is a source of multiple halogenated compounds and, in a limited number of studies, has been shown to decrease enteric CH4 emission in vitro and in vivo. Similarly, oregano has been suggested as a potential CH4 mitigating agent. This study consisted of 2 in vitro and 2 in vivo experiments. Experiment (Exp.) 1 was aimed at establishing the effect of AT on CH4 emission in vitro. Two experiments (Exp. 2 and 3) with lactating dairy cows were conducted to determine the antimethanogenic effect of AT and oregano (Exp. 3) in vivo. Another experiment (Exp. 4) was designed to investigate stability of bromoform (CHBr3) in AT over time. In Exp. 3, 20 Holstein cows were used in a replicated 4 × 4 Latin square design with four 28-d periods. Treatments were basal diet (control) or basal diet supplemented with (dry matter basis) 0.25% AT (LowAT), 0.50% AT (HighAT), or 1.77% oregano (Origanum vulgare L.) leaves. Enteric gas emissions were measured using the GreenFeed system (C-Lock Inc., Rapid City, SD), and rumen samples were collected for fermentation analysis using the ororuminal technique. In Exp.1 (in vitro), relative to the control, AT (at 1% dry matter basis, inclusion rate) decreased CH4 yield by 98%. In Exp. 3, HighAT decreased average daily CH4 emission and CH4 yield by 65% and 55%, respectively, in experimental periods 1 and 2, but had no effect in periods 3 and 4. The differential response to AT among experimental periods was likely a result of a decrease in CHBr3 concentration in AT over time, as observed in Exp. 4 (up to 84% decrease in 4 mo of storage). In Exp. 3, H2 emission was increased by AT and, as expected, the proportion of acetate in the total volatile fatty acids in the rumen was decreased and those of propionate and butyrate were increased by HighAT compared with the control. Compared with the control, HighAT decreased dry matter intake, milk yield, and energy-corrected milk yield in Exp. 3. Milk composition was not affected by treatment, except lactose percentage and yield were decreased by HighAT. Concentrations of iodine and bromide in milk were increased by HighAT compared with the control. Milk CHBr3 concentration and its organoleptic characteristics were not different between control and HighAT. Oregano had no effect on CH4 emission or lactational performance of the cows in Exp. 3. Overall, AT included at 0.50% in the ration of dairy cows can have a large mitigation effect on enteric CH4 emission, but dry matter intake and milk production may also decrease. There was a marked decrease in the CH4 mitigation potential of AT in the second half of Exp. 3, likely resulting from CHBr3 decay over time.

This study examined the effect of 3-nitrooxypropanol (3-NOP), an investigational substance, on enteric methane emission, milk production, and composition in Holstein dairy cows. Following a 3-wk covariate period, 48 multi- and primiparous cows averaging (± standard deviation) 118 ± 28 d in milk, 43.4 ± 8 kg/d milk yield, and 594 ± 57 kg of body weight were blocked based on days in milk, milk yield, and enteric methane emission and randomly assigned to 1 of 2 treatment groups: (1) control, no 3-NOP, and (2) 3-NOP applied at 60 mg/kg feed dry matter. Inclusion of 3-NOP was through the total mixed ration and fed for 15 consecutive weeks. Cows were housed in a freestall barn equipped with a Calan Broadbent Feeding System (American Calan Inc., Northwood, NH) for monitoring individual dry matter intake and fed ad libitum once daily. Enteric gaseous emissions (methane, carbon dioxide, and hydrogen) were measured using 3 GreenFeed (C-Lock Inc., Rapid City, SD) units. Dry matter intake, cow body weight, and body weight change were not affected by 3-NOP. Compared with the control group, 3-NOP applied at 60 mg/kg feed dry matter decreased daily methane emission, emission yield, and emission intensity by 26, 27, and 29%, respectively. Enteric emission of carbon dioxide was not affected, and hydrogen emission was increased 6-fold by 3-NOP. Administration of 3-NOP had no effect on milk and energy-corrected milk yields and feed efficiency, increased milk fat and milk urea nitrogen concentrations, and increased milk fat yield but had no other effects on milk components. Concentration of C6:0 and C8:0 and the sum of saturated fatty acids in milk fat were increased by 3-NOP. Total trans fatty acids and the sum of polyunsaturated fatty acids were decreased by 3-NOP. In this experiment, 3-NOP decreased enteric methane daily emission, yield, and intensity without affecting dry matter intake and milk yield, but increased milk fat in high-producing dairy cows.

This study investigated the effects of feeding solvent-extracted canola meal (CM), extruded soybean meal (ESBM), or solvent-extracted soybean meal (SSBM) on an equivalent crude protein basis on performance, plasma AA profiles, enteric gas emissions, milk fatty acids, and nutrient digestibility in lactating dairy cows. Fifteen Holstein cows (95 ± 20 d in milk) were used in a replicated 3 × 3 Latin square design experiment with 3 periods of 28 d each. Treatments were 3 diets containing 17.1% CM, 14.2% ESBM, or 13.6% SSBM (dry matter basis). Vegetable oil was added (canola oil for CM or soybean oil for SSBM) to equalize the ether extract concentration of the diets. Rumen-protected Met was supplemented targeting digestible Met supply of 2.2% of metabolizable protein in all diets. Canola meal increased dry matter intake (DMI) by 5.9 and 8.9% in comparison with ESBM and SSBM, respectively. Milk urea nitrogen was lowest in CM, followed by SSBM, and was highest for ESBM. No differences were observed in feed efficiency, energy-corrected milk yield, and milk composition or component yields among treatments. Cows fed CM emitted less enteric CH4 per kg of DMI compared with both ESBM and SSBM, but CH4 emission intensity (CH4 per kg of energy-corrected milk) was similar among treatments. In summary, replacement of ESBM or SSBM with CM, on an equal crude protein basis, in the diet of lactating dairy cows enhanced DMI, but yields of energy-corrected milk and milk components and feed efficiency were similar among treatments.

The objective of this experiment was to compare ruminal fluid samples collected through rumen cannula (RC) or using an oral stomach tube (ST) for measurement of ruminal fermentation and microbiota variables. Six ruminally cannulated lactating Holstein cows fed a standard diet were used in the study. Rumen samples were collected at 0, 2, 4, 6, 8, and 12 h after the morning feeding on two consecutive days using both RC and ST techniques. Samples were filtered through two layers of cheesecloth and the filtered ruminal fluid was used for further analysis. Compared with RC, ST samples had 7% greater pH; however, the pattern in pH change after feeding was similar between sampling methods. Total volatile fatty acids (VFA), acetate and propionate concentrations in ruminal fluid were on average 23% lower for ST compared with RC. There were no differences between RC and ST in VFA molar proportions (except for isobutyrate), ammonia and dissolved hydrogen (dH2) concentrations, or total protozoa counts, and there were no interactions between sampling technique and time of sampling. Bacterial ASV richness was higher in ST compared with RC samples; however, no differences were observed for Shannon diversity. Based on Permanova analysis, bacterial community composition was influenced by sampling method and there was an interaction between sampling method and time of sampling. A core microbiota comprised of Prevotella, S24-7, unclassified Bacteroidales and unclassified Clostridiales, Butyrivibrio, unclassified Lachnospiraceae, unclassified Ruminococcaceae, Ruminococcus, and Sharpea was present in both ST and RC samples, although their relative abundance varied and was influenced by an interaction between sampling time and sampling method. Overall, our results suggest that ruminal fluid samples collected using ST (at 180 to 200 cm depth) are not representative of rumen pH, absolute values of VFA concentrations, or bacterial communities >2 h post-feeding when compared to samples of ruminal fluid collected using RC. However, ST can be a feasible sampling technique if the purpose is to study molar proportions of VFA, protozoa counts, dH2, and ammonia concentrations., Frontiers in Microbiology, 11, ISSN:1664-302X

The objective of this study was to evaluate whether pre-weaning heifer calves divergent for residual feed intake (RFI) or residual feed intake and body weight gain (RIG) exhibit differences in thermography, blood, and ruminal parameters. Thirty-two Gyr heifer calves were enrolled in a 63-d trial and classified into 2 feed efficiency (FE) groups based on RFI and RIG (mean ± 0.5 SD). The groups were classified as high efficiency (HE) RFI (HE RFI, n = 9), HE RIG (HE RIG, n = 10), low efficiency (LE) RFI (LE RFI, n = 10), and LE RIG (LE RIG, n = 11). The amount of whole milk provided for each calf was calculated based on their metabolic weight at birth (42% x BW0.75). The liquid diet was divided into two meals at 0700 and 1400 h. The total solid diet (TSD) was composed of 92% concentrate and 8% of Tifton 85 hay chopped in 5-cm lengths, as fed. Intake was measured daily. Blood concentrations of insulin, beta hydroxybutyrate, urea, and glucose, and ruminal pH, N-NH3, and volatile fatty acids (VFA) were evaluated at 14, 28, 42, 56, and 70 days of age. Thermal images of the calves were taken with an infrared camera (FLIR T420, FLIR Systems Inc., Wilsonville, OR) on d 56 (±3) at 0600 h, before the morning feeding. Total VFA concentration and propionate as % of total VFA were 24.2% and 22.2% lower in HE RFI compared to LE RFI calves, respectively. On the other hand, acetate as % of total VFA was 10.6% greater in HE RFI than LE RFI calves. Blood urea concentration tended to be greater in LE RFI than HE RFI calves. High efficiency HE RIG tended to have 6.8% greater acetate and 15.4% lower propionate as % of total VFA than LE RIG. Blood insulin concentration was greater and blood glucose tended to be greater for LE RIG than HE RIG group. Low efficiency RIG group had greater left rib, left flank, and anus surface temperature measured by infrared thermography than the HE RIG group. Differences in ruminal fermentation do not seem to be associated with pre-weaning calves efficiency, while differences in protein metabolism seem to affect RFI during this phase. Infrared thermography appears to be correlated to RIG in pre-weaning heifer calves.

The aims of this study were (1) to assess if there is phenotypical divergence for feed efficiency (FE) during the preweaning phase; (2) if FE is correlated with heat production (HP) measured by the face mask method or (3) by surface skin temperature via thermography, and (4) whether these methods are applicable to preweaned calves. Holstein × Gyr heifer calves (n = 36, birth body weight = 32.4 ± 6.6 kg) were enrolled and on trial between 4 and 12 wk of age and were classified into 2 residual feed intake (RFI) and residual body weight gain (RG) groups: high efficiency (HE; RFI, n = 10; and RG, n = 9) and low efficiency (LE; RFI, n = 10; and RG, n = 8). Calves were fed milk (6 L/d) and solid feed (95% starter and 5% chopped Tifton 85 hay, as fed). Growth was monitored weekly and feed intake (milk and solid feed) daily, during the whole period. Gas exchanges (O2 consumption and production of CO2 and CH4) were obtained using a face mask at 45 ± 5 d of age and HP was estimated. Maximum temperatures were measured at 7 sites with an infrared camera at 62 ± 7 d of age. There was divergence in RFI and RG. Respectively, HE and LE calves had RFI of -0.14 and 0.13 kg/d, and RG of 0.05 and -0.07 kg/d. Dry matter intake was 15% lower in HE-RFI compared with LE-RFI, but no differences were observed in average daily weight gain. Within the RG test, no differences were observed in dry matter intake or average daily gain. The HE-RFI calves consumed less O2 (L/d) and produced less CO2 (L/d). Heart rate and HP were lower for HE-RFI calves compared with LE-RFI. Residual feed intake was correlated with HP (r = 0.48), O2 consumption (r = 0.48), CO2 production (r = 0.48), and heart rate (r = 0.40). No differences were observed in HP and gas exchanges between RG groups. Methane production was null in both groups. Eye temperature measured by thermography was 0.5°C greater in HE-RG than LE-RG calves. Differences in skin temperature between HE and LE calves were not observed at the other sites. These results support the hypothesis that calves are divergent for RFI, RG, and FE during preweaning and divergence tests are applicable during this phase. The face mask method described here is a useful tool for estimating differences in HP among phenotypically divergent RFI calves. Eye temperature measured by infrared thermography may have potential to screen phenotypically divergent RG calves.

We aimed to determine the effects of partially replacing conventional solvent-extracted soybean meal by amino resin-treated soybean meal in the concentrate supplement of lactating dairy cows grazing Guinea grass (Panicum maximum cv. Mombaca) on dry matter intake, milk production and composition, feed and nitrogen use efficiency, and nutrient digestibility. Eighteen Holstein x Gyr crossbred multiparous cows (79 ± 38 days in milk and 37.4 ± 4.69 kg/d of milk yield) were used in a replicated 3 × 3 Latin square design study with three experimental periods of 21 d The solvent extract soybean meal of the control diet (CON) was replaced by 1.0 (TSBM1.0) and 1.5 (TSBM1.5) kg as-fed basis of amino resin-treated soybean meal (29 and 43% as dry-matter basis, respectively) in the treatment diets. Dry matter intake, milk casein, protein, and fat contents, feed efficiency, nitrogen use efficiency, and nutrient digestibility did not differ among treatments (P > 0.05). The metabolizable protein was not increased by the treatments; however, cows fed with TSBM1.5 tended to produce 1.2 kg/d more milk (P = 0.09) and 64.3 g/d more lactose (P = 0.06) than CON cows. Although the supply of metabolizable protein was not affected by treatments, increasing rumen undegradable protein supply by 43% by replacing conventional solvent-extracted soybean meal with amino resin-treated soybean meal tended to increase milk production of high producing grazing cows.

The present study aimed to evaluate the use of an internal dry period teat seal containing bismuth subnitrate (Teatseal®, Zoetis®, Florham Park, Nova Jersey, USA) associated with a long-acting cloxacilin preparation (Orbenin® Extra dry cow, Zoetis®, Florham Park, Nova Jersey, USA), in preventing new infections during the dry-off and early postpartum period. A total of 150 Holstein cows (average production of 9,000 kg of milk per lactation), with four functional udder quarters without clinical mastitis was included in the study. All animals were dried-off 60 days before the expected calving date. Two teats positioned diagonal-contralaterally received only dry cow antibiotic, control group C (n=300) and the other two teats, treatment group T (n=300) received dry cow antibiotic and infusion with an internal teat seal. Data from SCC variable were transformed by log base-10 transformation. Duncan’s test was used accepting 5% as the level of statistical significance. The occurrence of intramammary infection (IMI) and chronicity rate, and frequency of microorganisms isolated at drying and immediately postpartum in teats of group C and group T were evaluated using a non-parametric Chi-square Test, accepting 10% as the statistical significance level. There was a decrease in the occurrence of new infections in the early postpartum in cows which the sealant was used (C=19.6%, T=11.4%). In the postpartum period, Gram-negative bacteria were isolated from 16 teats in C and seven in T. The greatest reduction was observed for Escherichia coli (8 vs 1) in group T. There was no effect using the internal sealant on the frequency of isolation of environmental Streptococus. The use of sealant reduced the prevalence of subclinical mastitis cows between drying-off and the early postpartum period (C=51% versus T=42%) and resulted in a lower somatic cell count (SCC) in the treatment group when compared with the control group (T=1,073x103, C=1,793x103). The use of the internal teat seal combined with dry cow antibiotic is effective in the prevention of IMI during the dry period and early lactation and results in the reduction of SCC in immediate postpartum period. The treatment is effective in reducing infection between dry-off and the immediate postpartum caused by major and minor pathogens. However, no effect on infections caused by contagious pathogens was observed. RESUMO: Objetivou-se avaliar o uso de um selante interno de tetos contendo subnitrato de bismuto (Teatseal®, Zoetis®, Florham Park, Nova Jersey, EUA) associado a um antibiótico de longa ação contendo cloxacilina (Orbenin® Extra dry cow, Zoetis®, Florham Park, Nova Jersey, EUA) em prevenir novas infecções durante o período seco e pós-parto imediato. Foram utilizadas 150 vacas Holandesas (produção média de 9,000 kg de leite por lactação), com os quatro quartos mamários funcionais e sem mastite clínica, que foram secas 60 dias antes da data prevista para o parto. O teto constituiu a unidade experimental. O grupo controle (C) foi representado por dois tetos diagonais-contralaterais (n=300), que receberam somente o antibiótico de vaca seca. Os outros dois tetos (n=300) constituíam o grupo tratado (T) e recebiam o antibiótico de vaca seca associado ao selante interno. Dados da variável contagem de células somáticas (CCS) passaram por transformação logarítmica na base 10 e foram submetidos ao teste de Duncan, aceitando-se nível de 5% de significância estatística. A ocorrência de infecções intramamárias (IIM), taxa de cronicidade e frequência de microrganismos isolados na secagem e imediatamente pós-parto nos tetos do grupo controle e tratados com selante interno foram avaliados utilizando o teste não paramétrico qui-quadrado, aceitando 10% de nível de significância. Houve redução na ocorrência de novas infecções no pós-parto imediato nos animais em que se utilizou o selante (C=19,6%, T=11,4%). No período pós-parto, foram isolados microrganismos Gram-negativo de 16 tetos no C e sete no T. A maior redução ocorreu para Escherichia coli, (8 versus 1) no grupo T. Não houve efeito da utilização do selante interno na frequência de isolamento de Streptococcus ambientais. O uso do selante reduziu a prevalência de mastite subclínica das vacas no pós-parto imediato (C=51% versus T=42%) e resultou em menor CCS no grupo tratado (T = 1.073 x 103) quando comparado com o grupo controle (C=1.793x103). O uso do selante interno combinado com antibiótico de vacas secas é eficaz na prevenção de infecções intramamárias no período seco e início de lactação e resulta na redução da CCS no período pós-parto imediato. O tratamento é eficaz na redução de infecção entre a secagem e o pós-parto imediato causada por patógenos maiores e menores. Entretanto, não tem efeito sobre infecções causadas por patógenos contagiosos.

The objective of this study was to assess the control feasibility of Streptococcus agalactiae subclinical mastitis and its impact in reducing somatic cell counts (SCC) of bulk milk tank. During the lactation, 33 cows positive for Streptococcus agalactiae in microbiological culture were treated with three applications of intramammary antimicrobial containing 100.000 UI of penicillin and 150 mg of novobiocin, in an interval of 12 hours. Individual and bulk tank milk samples were collected for somatic cell count (SCC) analysis and milk yield was measured before and after treatment. Statistical analyses were performed with SAS System, version 9.2. All cows treated against S. agalactiae had microbiological cure and reduction of SCC, indicating the effectiveness of treatment. SCC in bulk tank was reduced from 829,000 to 513,000 cells/ml.

The objectives of this study were: 1) to classify animals into groups of high and low feed efficiency using two feed efficiency indexes (Residual feed intake (RFI) and residual feed intake and body weight gain (RIG)), and 2) to evaluate if pre-weaning heifer calves divergent for feed efficiency indexes exhibit differences in performance, body measurements, digestibility, energy partitioning, and nitrogen partitioning. A total of 32 Gyr heifer calves were enrolled in a 63-d trial and classified into two feed efficiency (FE) groups based on RFI and RIG (mean ± 0.5 SD). The groups were classified as high efficiency (HE) RFI (HE RFI, n = 9; HE RIG, n = 10), and low efficiency (LE) RFI (LE RFI, n = 10; LE RIG, n = 11). The remaining animals were classified as intermediate (n = 13 (RFI) and n = 11 (RIG)). HE and LE calves had RFI values of-0.052 and 0.049 kg/d (P < 0.05), respectively. The HE RFI group consumed 8.9% less solid diet than the LE RFI group. HE RFI animals exhibited an increased digestibility of crude protein and ether extract and tended to have greater total dry and organic matter digestibility. LE RFI animals had greater gross energy and nitrogen intake, though greater fecal losses resulted in a tendency to reduce energy and nitrogen use efficiency. HE and LE calves had RIG values of 0.080 and -0.077kg/d (P ≤ 0.01), respectively. HE RIG animals exhibited greater average daily gain (9.4%), body weight (BW), and heart girth, though HE RIG group exhibited narrower hip width. HE RIG animals tended to have greater ether extract digestibility but greater methane losses (% of gross energy). HE RFI in pre-weaning heifers seems to be related to differences in digestibility. Divergent animals for RIG during the assessed phase appear to differ in body measurements, which may be related to differences in the composition of the gain.

The recent advances in sensor technology have allowed accurate predictions of estrus events using animal behavior information. Behavioral variables generated by electronic feed and water bins have not been explored as potential predictors for estrus detection. The objectives of this study were: (i) to evaluate the effect of estrus expression on feed intake and animal behavior (feeding and drinking) and (ii) to develop and evaluate predictive approaches to detect estrus expression using electronic feed and water bins data. Feed intake, animal behavior, and estrus events were measured in 57 Holstein × Gyr heifers (374 ± 21.2 kg and 22.6 ± 0.60 months). Previous to each estrus event, the following covariates were computed: total feed intake (FI, as-fed basis), number of visits at the feed bins (VF) and water bins (VW), time spent eating (TE), and time spent drinking water (TD). Three predictive approaches were evaluated: Generalized Linear Models (GLM), Artificial Neural Network (ANN), and Random Forest (RF). Twelve covariate sets were established to investigate: (ii.a) the prediction quality for estrus detection when long (−174 to 0 h) or short (−24 to 0 h) time series were used as predictors (6 h of time window, with estrus event at 0 h); (ii.b) the ability of machine learning algorithms to predict estrus 6 and 12 h in advance; and (ii.c) the predictive quality for estrus detection when only feeding and drinking behavior data (without intake variables) were included as predictors. The predictive approaches were evaluated through Leave-One-Out Cross-validation. Estrus events altered feeding and drinking behavior patterns, and feed intake. ANN, RF, and GLM presented similar accuracies within covariate sets. There was no benefit of using longer time series for estrus detection. Earlier detection of estrus event (6 and 12 h in advance) reduced model accuracy compared to predictions performed at 0 h. However, ANN and RF showed accuracy values ranging between 75.7% and 96.5%, which indicates a great potential for early estrus detection. The exclusion of feed intake data of the covariate sets did not reduce the accuracy, sensitivity, and specificity of the models for estrus detection. These findings suggest that behavioral data can early predict estrus events, which could be incorporated in sensor technologies capable of generating behavioral information, such as electronic bins, wearable sensors, and computer vision systems.

RESUMO Contagem de células somáticas (CCS) e produção de leite de vacas mestiças Holandês x Gir foram utilizadas para avaliação da perda de produção de leite estimada por duas metodologias, baseadas na CCS média da lactação total encerrada ou na CCS e nas pesagens de leite em diferentes estágios de lactação. Nas primíparas consideradas com mastite subclínica (CCS ≥ 200.000 células mL-1), a perda de produção na lactação total, avaliada pela CCS média da lactação, foi de 814kg. Para as multíparas, não houve diferença na produção de leite entre vacas com e sem MSC com base na CCS média da lactação. A estimativa avaliada por meio de testes mensais de CCS e pesagens de leite em diferentes estágios de lactação, balanceada pela prevalência de mastite subclínica do mês foi de 917,9kg para primíparas e 1178,9kg para multíparas. A contraposição da produção de leite com a CCS em diferentes estágios da lactação evidenciou maior perda de produção em relação à observação da CCS média da lactação. A perda de produção de leite de vacas baseada na CCS média de toda a lactação foi subestimada quando comparada à contraposição da produção de leite com CCS em diferentes estágios da lactação. ABSTRACT Somatic cell count (SCC) and milk production of crossbred Holstein x Gir were used to evaluate the estimated loss of milk production two methodologies, based on the average SCC of total closed lactation, or average SCC and weighing milk at different stages of lactation. In primiparous with SCC mL ≥200,000 cells-1 production loss in the total lactation, evaluated by the average lactation SCC, was 814 kilograms. For multiparous, there was no difference in milk production between cows with and without SCM based on the SCC media of lactation. The estimate of lost production assessed through monthly testing of SCC and milk weighing at different stages of lactation, balanced by the prevalence of subclinical mastitis, was 917.9kg for primiparous and 1178.9kg for multiparous. The estimative of milk production with CCS at different stages of lactation showed greater loss of milk production when compared to the estimative based on the average SCC throughout the period of lactation. The milk production loss method based on the average somatic cell count of the whole lactation was underestimated when compared to the method based on average SCC in different stages of lactation.

The aim of this study was to evaluate the effects of increasing the total solids (TS) content of liquid feed, by adding increasing amounts of milk replacer powder to whole milk, on age at puberty and postweaning performance, glucose metabolism, and mammary fat pad of dairy heifers. Crossbreed Holstein × Gyr heifers (n = 58) were distributed into 4 different treatments during the preweaning period. Treatments consisted of liquid feeds with TS content of 12.5 (actual TS = 13.5 ± 0.53%; n = 15), 15.0 (actual TS = 16.1 ± 0.03%; n = 15), 17.5 (actual TS = 18.2 ± 0.14%; n = 13), or 20.0% (actual TS = 20.4 ± 0.24%; n = 15), which were provided up to 59 d old. From 60 to 89 d old, the animals received the same starter offered during the preweaning period in addition to corn silage ad libitum starting at 70 d old. From 90 to 104 d old, the animals were adapted to a total mixed ration. At 105 d old, the animals were distributed in 4 paddocks equipped with electronic feed and water bins and were fed the same total mixed ration ad libitum. A period of adaptation to the electronic feed bins occurred from 105 to 119 d old. Feed and water intake and body weight and body frame development were assessed until puberty. Mammary gland evaluations were performed monthly by ultrasonography from 120 d of age until puberty onset. Puberty onset was determined as plasma progesterone concentration greater than 1 ng/mL in 2 consecutive samples collected 7 d apart. The date of puberty onset was recorded as the collection day of the first of these samples. A glucose tolerance test was performed at 280 d of age. The increased TS content of the liquid feed fed during the preweaning period did not affect dry matter intake, performance, age at puberty, glucose metabolism, or mammary gland fat pad at later stages of rearing process. Conversely, hip width and heart girth increased linearly up to 150 and 240 d of age, respectively, as a result of the increased TS concentration of the liquid feed during the preweaning period.

RESUMO: Aproximadamente 75% das doenças em vacas leiteiras acontecem no primeiro mês após o parto e esses problemas têm sua origem associada ao sistema imune e ao consumo de alimentos duas a três semanas antes do parto. Doenças relacionadas ao trato reprodutivo como, por exemplo, retenção de placenta pode afetar a eficiência reprodutiva assim como a produção de leite. Os efeitos das doenças nos processos fisiológicos dos animais se transformam em impactos econômicos passíveis de serem mensurados. Objetivou-se avaliar o impacto econômico da retenção de placenta em um rebanho composto de 900 vacas em lactação. O diagnóstico de retenção de placenta foi definido como presença da placenta 24 horas após o parto. Foi utilizado um banco de dados no estudo, do qual foram extraídas as seguintes informações: ano de parição, época de parição, ordem de lactação, presença da placenta 24 horas após o parto, e número de doses de sêmen por gestação. Para cálculo do impacto econômico foram considerados os custos diretos (tratamento, mão de obra, redução da produção de leite e descarte de leite durante o período de tratamento) e indiretos (aumento do período de serviço, aumento do número de doses de sêmen e aumento do risco de descarte). Os valores médios relacionados aos custos foram obtidos junto ao veterinário responsável pela propriedade com referência ao ano de 2009 para que os cálculos fossem realizados. O custo total por ocorrência de retenção de placenta em vacas primíparas no período de chuva e seca foi de US$51,8 e em vacas multíparas foi de US$70,6 e US$87,9 nas épocas de seca e de chuvas, respectivamente. A retenção de placenta apresentou custo para a propriedade no período avaliado de US$8.878,0 ou 19.666 litros de leite. A retenção de placenta apresentou impacto econômico importante na propriedade leiteira nas condições avaliadas. ABSTRACT: Approximately 75% of diseases in dairy cows occur in the first month after parturition and these problems have their origin associated with the immune system and food consumption two to three weeks before parturition. Diseases related to the reproductive tract, such as retained placenta, can affect reproductive efficiency as well as milk production. The effects of diseases on the physiological processes of animals become economic impacts that can be measured. The objective of this study was to evaluate the economic impact of retained placenta on a herd of 900 lactating cows. The diagnosis of placental retention was defined as presence of the placenta 24 hours postpartum. A database was used in the study, from which the following information was extracted: placenta presence 24 hours after calving, year of calving, calving season, and number of semen doses per gestation. In order to calculate the economic impact, the direct costs (treatment, labor, reduction of milk production and milk discharge during the treatment period) and indirect costs (increase in the service period, increase in the number of semen doses and increase of the risk of disposal). The average values related to the costs were obtained from the veterinarian responsible for the property with reference to the year 2009 for the calculations to be carried out. The total cost per occurrence of placenta retention in primiparous cows in the rainy and dry season was US$51.8 and in multiparous cows was US$70.6 and US$87.9 in times of drought and rain, respectively. Retention of placenta presented a cost for the property in the evaluated period of US$8,878.0 or 19,666 liters of milk. Retention of placenta presented a significant economic impact on milk production under the conditions evaluated.