Articles in Press

Short communication: Comparison of pH, volatile fatty acids, and microbiome of rumen samples from preweaned calves obtained via cannula or stomach tube - Corrected Proof
M. Terré, L. Castells, F. Fàbregas, A. Bach

ABSTRACT: The objective of this study was to compare rumen samples from young dairy calves obtained via a stomach tube (ST) or a ruminal cannula (RC). Five male Holstein calves (46 ± 4.0 kg of body weight and 11 ± 4.9 d of age) were ruminally cannulated at 15 d of age. Calves received 4 L/d of a commercial milk replacer (25% crude protein and 19.2% fat) at 12.5% dry matter, and were provided concentrate and chopped oats hay ad libitum throughout the study (56 d). In total, 29 paired rumen samples were obtained weekly throughout the study in most of the calves by each extraction method. These samples were used to determine pH and volatile fatty acids (VFA) concentration, and to quantify Prevotella ruminicola and Streptococcus bovis by quantitative PCR. Furthermore, a denaturing gradient gel electrophoresis was performed on rumen samples harvested during wk 8 of the study to determine the degree of similarity between rumen bacteria communities. Rumen pH was 0.30 units greater in ST compared with RC samples. Furthermore, total VFA concentrations were greater in RC than in ST samples. However, when analyzing the proportion of each VFA by ANOVA, no differences were found between the sampling methods. The quantification of S. bovis and P. ruminicola was similar in both extraction methods, and values obtained using different methods were highly correlated (R2 = 0.89 and 0.98 for S. bovis and P. ruminicola, respectively). Fingerprinting analysis showed similar bacteria band profiles between samples obtained from the same calves using different extraction methods. In conclusion, when comparing rumen parameters obtained using different sampling techniques, it is recommended that VFA profiles be used rather than total VFA concentrations, as total VFA concentrations are more affected by the method of collection. Furthermore, although comparisons of pH across studies should be avoided when samples are not obtained using the same sampling method, the comparison of fingerprinting of a bacteria community or a specific rumen bacterium is valid.

Efficacy of extended cefquinome treatment of clinical Staphylococcus aureus mastitis - Corrected Proof
J.M. Swinkels, P. Cox, Y.H. Schukken, T.J.G.M. Lam

ABSTRACT: Clinical Staphylococcus aureus mastitis is difficult to cure. Extended antimicrobial treatment is often advocated as a practical approach to improve cure rates; however, scientific evidence of this hypothesis is lacking. A multi-centered, nonblinded, randomized, positive-controlled clinical trial was conducted in 5 European countries—France, Hungary, Italy, the Netherlands, and the United Kingdom—to study the efficacy of an extended intramammary cefquinome treatment (5 d) compared with a standard intramammary cefquinome treatment (1.5 d) of Staph. aureus clinical mastitis. Least squares means estimates of bacteriological cure during lactation were 34% [standard error (SE) = 9.9%] for the standard treatment group and 27% (SE = 8.4%) for the extended treatment group. In the final model, extended therapy was not significantly better. The only factor predicting bacteriological cure was pretreatment cow somatic cell count (SCC). Cows with >250,000 cells/mL in milk before treatment were less likely to cure. Least squares means of clinical cure during lactation was 60% (SE = 19%) for the standard treatment group and 82% (SE = 12%) for the extended treatment group. In the final model, clinical cure after extended treatment was significantly better. Pretreatment cow udder firmness predicted clinical cure. Firm udders were less likely to cure clinically. Irrespective of treatment regimen, new infection rates with pathogens other than Staph. aureus were higher (42%) after bacteriological cure than after nonbacteriological cure (22%) and cured cows had a significantly lower SCC. In conclusion, independent of the treatment protocol, cows with an SCC <250,000 cells/mL before treatment showed a higher probability of bacteriological cure. It appears that successful treatment of clinical Staph. aureus mastitis with cefquinome is associated with an increased number of new infections with coagulase-negative staphylococci. Extended treatment improved clinical, but not bacteriological, cure rates compared with the standard treatment. These results indicate that extending treatment of clinical Staph. aureus mastitis with cefquinome should not be recommended.

Feasibility of using automatic milking system data from commercial herds for genetic analysis of milkability - Corrected Proof
C. Carlström, G. Pettersson, K. Johansson, E. Strandberg, H. Stålhammar, J. Philipsson

ABSTRACT: The objective of this study was to investigate how useful data from automatic milking systems used in commercial herds are for genetic analysis of milkability traits. Data were available from 4,968 Swedish Holstein and Swedish Red cows over a span of 5 yr (2004–2009) from 19 herds. The analyzed milkability traits were average flow rate, box time, milking interval, and number of milkings per day. Variance components were estimated for genetic, permanent environmental, and residual effects in first and later (second and third) lactations, and were used for estimation of heritabilities and repeatablilites. The experiences of the data quality and editing procedures showed that almost half of the data and about a quarter of the cows had to be excluded from the analyses due to incomplete or inconsistent information. However, much more data are available than is needed for accurate genetic parameter estimations. For the genetic analysis, a repeatability animal model was used that included the fixed effects of herd, year and season, lactation month, and milk yield. The repeatability coefficients were at a high level: highest for average flow rate, with estimates between 0.8 and 0.9. The estimated heritability coefficients were in the range of 0.37 to 0.48, 0.21 to 0.44, 0.09 to 0.26, and 0.02 to 0.07 for average flow rate, box time, milking interval, and number of milkings, respectively. The results from the present study unraveled large genetic variation in milkability traits. The genetic parameter estimates were well in agreement with previous studies of milkability, which proves the feasibility of using data from automatic milking systems for genetic analysis.

What do preweaned and weaned calves need in the diet: A high fiber content or a forage source? - Corrected Proof
M. Terré, E. Pedrals, A. Dalmau, A. Bach

ABSTRACT: The objective of this study was to determine whether the improvement of performance of young calves associated with the supplementation of chopped grass hay reported in some studies is due to an increase in the total neutral detergent fiber (NDF) content of the consumed diet or to the provision of chopped grass hay. Sixty-three Holstein calves [9 ± 4.4 d old; mean ± standard deviation (SD)] were randomly distributed in 4 treatments resulting from the combination of 2 levels of NDF content of a pelleted starter and the supply or absence of forage provision: low-NDF starter (18%) with or without chopped oat hay, and high-NDF starter (27%) with or without chopped oat hay. All animals were fed the same milk replacer (21% crude protein and 19.2% fat) at the rate of 4 L/d at 15% dry matter from d 1 to 34, and 2 L/d at 15% dry matter from d 35 to 42 (weaning). The study finished 2 wk after weaning. Body weight was measured weekly and individual calf starter and hay intake was recorded daily. On d 50, blood samples were drawn 2 h after the morning concentrate offer to determine serum glucose and insulin concentrations. On d 52, samples of ruminal fluid were obtained via an esophageal tube, and pH was measured immediately. During the preweaning period, pelleted starter intake was similar among treatments, but average daily gain tended to be greater in low- than in high-NDF treatments (0.69 vs. 0.63 ± 0.020 kg/d, respectively; mean ± SD). However, during the 2 wk after weaning, supplementation of forage improved pelleted starter intake and average daily gain without affecting the gain-to-feed ratio. Probably, the greater pelleted starter intake observed in forage-supplemented calves was mainly due to the greater ruminal pH found in forage-supplemented calves compared with forage-deprived calves (5.81 vs. 5.05 ± 0.063, respectively). Blood insulin-to-glucose ratio was greater in forage-supplemented compared with unsupplemented calves [mean ± SD; 6.53 vs. 4.24 ± 0.125 insulin (ng/L)-to-glucose (mg/dL) ratio, respectively]. In conclusion, a low-NDF pelleted starter is recommended during the preweaning period, and the provision of chopped hay is necessary right after weaning to improve calf performance.

Physical properties of pizza Mozzarella cheese manufactured under different cheese-making conditions - Corrected Proof
V. Banville, P. Morin, Y. Pouliot, M. Britten

ABSTRACT: The effect of manufacturing factors on the shredability and meltability of pizza Mozzarella cheese was studied. Four experimental cheeses were produced with 2 concentrations of denatured whey protein added to milk (0 or 0.25%) and 2 renneting pH values (6.4 or 6.5). The cheeses were aged 8, 22, or 36 d before testing. Shredability was assessed by the presence of fines, size of the shreds, and adhesion to the blade after shredding at 4, 13, or 22°C. A semi-empirical method was developed to measure the matting behavior of shreds by simulating industrial bulk packaging. Rheological measurements were performed on cheeses with and without a premelting treatment to assess melt and postmelt cheese physical properties. Lowering the pH of milk at renneting and aging the cheeses generally decreased the fines production during shredding. Adding whey protein to the cheeses also altered the fines production, but the effect varied depending on the renneting and aging conditions. The shred size distribution, adhesion to the blade, and matting behavior of the cheeses were adversely affected by increased temperature at shredding. The melting profiles obtained by rheological measurements showed that better meltability can be achieved by lowering the pH of milk at renneting or aging the cheese. The premelted cheeses were found to be softer at low temperatures (<40°C) and harder at high temperatures (>50°C) compared with the cheeses that had not undergone the premelting treatment. Understanding and controlling milk standardization, curd acidification, and cheese aging are essential for the production of Mozzarella cheese with desirable shredability and meltability.

Influence of packaging information on consumer liking of chocolate milk - Corrected Proof
M.K. Kim, K. Lopetcharat, M.A. Drake

ABSTRACT: Chocolate milk varies widely in flavor, color, and viscosity, and liking is influenced by these properties. Additionally, package labels (declared fat content) and brand are some of the extrinsic factors that may influence consumer perception. The objective of this study was to evaluate the effects of packaging labels and brand name on consumer liking and purchase intent of chocolate milk. A consumer acceptance test, conjoint analysis survey, and Kano analysis were conducted. One hundred-eight consumers evaluated 7 chocolate milks with and without brand or package information in a 2-d crossover design. A conjoint analysis survey and Kano analysis were conducted after the consumer acceptance test. Results were evaluated by 2-way ANOVA and multivariate analyses. Declared fat content and brand influenced overall liking and purchase intent for chocolate milks to differing degrees (P < 0.05). A subsequent conjoint analysis (n = 250) revealed that fat content was a driver of choice for purchasing chocolate milk followed by sugar content and brand. Brand name was less important for purchase intent of chocolate milk than fat or sugar content. Among fat content of chocolate milk, 2 and 1% fat level were most appealing to consumers, and reduced sugar and regular sugar were equally important for purchase intent. Kano analysis confirmed that fat content (whole milk, 1, or 2% fat chocolate milk) was an attractive attribute for consumer satisfaction, more so than brand. Organic labeling did not affect the purchase decision of chocolate milk; however, Kano results revealed that having an organic label on a package positively influenced consumer satisfaction. Findings from this study can help chocolate milk producers as well as food marketers better target their product labels with attributes that drive consumer choice of chocolate milk.

Proteolysis and biogenic amine buildup in high-pressure treated ovine milk blue-veined cheese - Corrected Proof
J. Calzada, A. Del Olmo, A. Picon, P. Gaya, M. Nuñez

ABSTRACT: Penicillium roqueforti plays an important role in the ripening of blue-veined cheeses, mostly due to lactic acid consumption and to its extracellular enzymes. The strong activity of P. roqueforti proteinases may bring about cheese over-ripening. Also, free amino acids at high concentrations serve as substrates for biogenic amine formation. Both facts result in shorter product shelf-life. To prevent over-ripening and buildup of biogenic amines, blue-veined cheeses made from pasteurized ovine milk were high-pressure treated at 400 or 600 MPa after 3, 6, or 9 wk of ripening. Primary and secondary proteolysis, biogenic amines, and sensory characteristics of pressurized and control cheeses were monitored for a 90-d ripening period, followed by a 270-d refrigerated storage period. On d 90, treatments at 400 MPa had lowered counts of lactic acid bacteria and P. roqueforti by less than 2 log units, whereas treatments at 600 MPa had reduced lactic acid bacteria counts by more than 4 log units and P. roqueforti counts by more than 6 log units. No residual α-casein (CN) or κ-CN were detected in control cheese on d 90. Concentrations of β-CN, para-κ-CN, and γ-CN were generally higher in 600 MPa cheeses than in the rest. From d 90 onwards, hydrophilic peptides were at similar levels in pressurized and control cheeses, but hydrophobic peptides and the hydrophobic-to-hydrophilic peptide ratio were at higher levels in pressurized cheeses than in control cheese. Aminopeptidase activity, overall proteolysis, and free amino acid contents were generally higher in control cheese than in pressurized cheeses, particularly if treated at 600 MPa. Tyramine concentration was lower in pressurized cheeses, but tryptamine, phenylethylamine, and putrescine contents were higher in some of the pressurized cheeses than in control cheese. Differences in sensory characteristics between pressurized and control cheeses were generally negligible, with the only exception of treatment at high pressure level (600 MPa) at an early ripening stage (3 wk), which affected biochemical changes and sensory characteristics.

Effects of forage provision to young calves on rumen fermentation and development of the gastrointestinal tract - Corrected Proof
L. Castells, A. Bach, A. Aris, M. Terré

ABSTRACT: Fifteen Holstein male calves were randomly assigned to 1 of 3 dietary treatments according to age and body weight (BW) to determine the effects of feeding different forages sources on rumen fermentation and gastrointestinal tract (GIT) development. Treatments consisted of a starter (20% crude protein, 21% neutral detergent fiber) fed alone (CON) or supplemented with alfalfa (AH) or with oat hay (OH). All calves received 2 L of milk replacer (MR) at 12.5% dry matter twice daily until 49 d of age. Calves received 2 L of the same MR from 50 to 56 d of age and were weaned at 57 d of age. Individual starter, forage, and MR intakes were recorded daily and BW was recorded weekly. A rumen sample was taken weekly to determine rumen pH and volatile fatty acid concentrations. Three weeks after weaning, animals were harvested and each anatomical part of the GIT was separated and weighed with and without contents. Rumen pH was lower in CON than in OH and AH calves. Furthermore, acetate proportion in the rumen liquid tended to be greater in AH than in CON and OH treatments. Total GIT weight, expressed as a percentage of BW, tended to be greater in AH compared with the other 2 treatments. Rumen tissue tended to weigh more in CON than in OH animals. Animals with access to forage tended to have a greater expression of monocarboxylate transporter 1 than CON calves. In conclusion, calves supplemented with oat hay have a better rumen environment than calves offered no forage and do not have an increased gut fill.

Newly identified mutations at the CSN1S1 gene in Ethiopian goats affect casein content and coagulation properties of their milk - Corrected Proof
T.A. Mestawet, A. Girma, T. Ådnøy, T.G. Devold, G.E. Vegarud

ABSTRACT: Very high casein content and good coagulation properties previously observed in some Ethiopian goat breeds led to investigating the αs1-casein (CSN1S1) gene in these breeds. Selected regions of the CSN1S1 gene were sequenced in 115 goats from 5 breeds (2 indigenous: Arsi-Bale and Somali, 1 exotic: Boer, and 2 crossbreeds: Boer × Arsi-Bale and Boer × Somali). The DNA analysis resulted in 35 new mutations: 3 in exons, 3 in the 5′ untranslated region (UTR), and 29 in the introns. The mutations in exons that resulted in an amino acid shift were then picked to evaluate their influence on individual casein content (αs1-, αs2-, β-, and κ-CN), micellar size, and coagulation properties in the milk from the 5 goat breeds. A mutation at nucleotide 10657 (exon 10) involved a transversion: CAG→CCG, resulting in an amino acid exchange Gln77→Pro77. This mutation was associated with the indigenous breeds only. Two new mutations, at nucleotide 6072 (exon 4) and 12165 (exon 12), revealed synonymous transitions: GTC→GTT in Val15 and AGA→AGG in Arg100 of the mature protein. Transitions G→A and C→T at nucleotides 1374 and 1866, respectively, occurred in the 5′ UTR, whereas the third mutation involved a transversion T→G at nucleotide location 1592. The goats were grouped into homozygote new (CC), homozygote reference (AA), and heterozygote (CA) based on the nucleotide that involved the transversion. The content of αs1-CN (15.32 g/kg) in milk samples of goats homozygous (CC) for this newly identified mutation, Gln77→Pro77 was significantly higher than in milks of heterozygous (CA; 9.05 g/kg) and reference (AA; 7.61 g/kg) genotype animals. The αs2-, β-, and κ-CN contents showed a similar pattern. Milk from goats with a homozygous new mutation had significantly lower micellar size. Milk from both homozygote and heterozygote new-mutation goats had significantly shorter coagulation rate and stronger gel than the reference genotype. Except the transversion, the sequence corresponded to allele A and presumably derived from it. Therefore, this allele is denoted by A3. All goats from the reference genotype (AA) were homozygous for the allele at nucleotide position 1374 and 1866, whereas all mutations in the 5′ UTR existed in a heterozygous form in both heterozygous (CA) and the new mutation (CC) genotype. The newly identified mutation (CC) detected in some of the goat breeds is, therefore, important in selection for genetic improvement and high-quality milk for the emerging goat cheese-producing industries. The finding will also benefit farmers raising these goat breeds due to the increased selling price of goats. Further studies should investigate the effect of this amino acid exchange on the secondary and tertiary structure of the αs1-CN molecule and on the susceptibility of peptide hydrolysis by digestive enzymes.

Risk factors for clinical mastitis, ketosis, and pneumonia in dairy cattle on organic and small conventional farms in the United States - Corrected Proof
R.M. Richert, K.M. Cicconi, M.J. Gamroth, Y.H. Schukken, K.E. Stiglbauer, P.L. Ruegg

ABSTRACT: The US regulations for production of organic milk include a strict prohibition against the use of antimicrobials and other synthetic substances. The effect of these regulations on dairy animal health has not been previously reported. The objective of this study was to characterize disease detection and identify risk factors for selected diseases on organic (ORG) and similarly sized conventional (CON) farms. Dairy herds (n = 292) were enrolled across 3 states (New York, Oregon, Wisconsin) with CON herds matched to ORG herds based on location and herd size. During a single herd visit, information was collected about herd management practices and animal disease occurring in the previous 60 d, and paperwork was left for recording disease occurrences during 60 d after the visit. For analysis, CON herds were further divided into grazing and nongrazing. Poisson regression models were used to assess risk factors for rate of farmer-identified and recorded cases of clinical mastitis, ketosis, and pneumonia. An increased rate of farmer-identified and recorded cases of clinical mastitis was associated with use of CON management, use of forestripping, presence of contagious pathogens in the bulk tank culture, proactive detection of mastitis in postpartum cows, and stall barn housing. An increased rate of farmer-identified and recorded cases of ketosis was associated with having a more sensitive definition of ketosis, using stall barn housing, and feeding a greater amount of concentrates. An increased rate of farmer-identified and recorded cases of pneumonia was associated with a lack of grazing, small or medium herd size, and Jersey as the predominant breed. Overall, disease definitions and perceptions were similar among grazing systems and were associated with the rate of farmer-identified and recorded cases of disease.

Short communication: Effects of level of rumen-degradable protein and corn distillers grains in corn silage-based diets on milk production and ruminal fermentation in lactating dairy cows - Corrected Proof
G.I. Zanton, A.J. Heinrichs, C.M. Jones

ABSTRACT: Two of the potential obstacles precluding inclusion of higher levels of dry distillers grains with solubles (DDGS) in corn-based dairy cow diets are the low levels of rumen-degradable protein (RDP) and the fatty acid content and composition of DDGS. Therefore, the objective of this experiment was to evaluate the production and rumen responses to dietary alterations in the level of RDP and DDGS for dairy cows fed a high corn silage diet. The experimental design was a replicated 4 × 4 Latin square with 21-d periods: 14 d of adaptation and 7 d of sampling; 16 uncannulated cows and 4 ruminally cannulated cows were blocked and assigned randomly to treatment sequences. Rations were provided as total mixed rations and were formulated to be high or low in RDP, with or without DDGS replacing soybean-based concentrates: high RDP, no DDGS (HRDP0); low RDP, no DDGS (LRDP0); low RDP, 10% DDGS; and low RDP, 20% DDGS. Body weight (696 kg) and dry matter intake (26.6 kg/d) were not affected by treatment. Rumen ammonia concentration was greater for HRDP0 than LRDP0, but was unaffected by level of DDGS inclusion. The mean and minimum rumen pH and time pH was <5.5 were not different between diets. Milk production tended to be lower for cows fed HRDP0 than LRDP0 and tended to be linearly reduced as DDGS inclusion increased. Milk protein yield tended to be greater for cows fed LRDP0 than HRDP0, but was unaffected by DDGS level. Milk fat production, concentration, and fat-corrected milk were linearly reduced by increasing levels of DDGS. Based on these results, feeding DDGS at 20% of diet dry matter is not recommended for diets based on high corn silage.

Lameness detection based on multivariate continuous sensing of milk yield, rumination, and neck activity - Corrected Proof
T. Van Hertem, E. Maltz, A. Antler, C.E B. Romanini, S. Viazzi, C. Bahr, A. Schlageter-Tello, C. Lokhorst, D. Berckmans, I. Halachmi

ABSTRACT: The objective of this study was to develop and validate a mathematical model to detect clinical lameness based on existing sensor data that relate to the behavior and performance of cows in a commercial dairy farm. Identification of lame (44) and not lame (74) cows in the database was done based on the farm's daily herd health reports. All cows were equipped with a behavior sensor that measured neck activity and ruminating time. The cow's performance was measured with a milk yield meter in the milking parlor. In total, 38 model input variables were constructed from the sensor data comprising absolute values, relative values, daily standard deviations, slope coefficients, daytime and nighttime periods, variables related to individual temperament, and milk session-related variables. A lame group, cows recognized and treated for lameness, to not lame group comparison of daily data was done. Correlations between the dichotomous output variable (lame or not lame) and the model input variables were made. The highest correlation coefficient was obtained for the milk yield variable (rMY = 0.45). In addition, a logistic regression model was developed based on the 7 highest correlated model input variables (the daily milk yield 4 d before diagnosis; the slope coefficient of the daily milk yield 4 d before diagnosis; the nighttime to daytime neck activity ratio 6 d before diagnosis; the milk yield week difference ratio 4 d before diagnosis; the milk yield week difference 4 d before diagnosis; the neck activity level during the daytime 7 d before diagnosis; the ruminating time during nighttime 6 d before diagnosis). After a 10-fold cross-validation, the model obtained a sensitivity of 0.89 and a specificity of 0.85, with a correct classification rate of 0.86 when based on the averaged 10-fold model coefficients. This study demonstrates that existing farm data initially used for other purposes, such as heat detection, can be exploited for the automated detection of clinically lame animals on a daily basis as well.

The relationship between endometrial cytology during estrous cycle and cutoff points for the diagnosis of subclinical endometritis in grazing dairy cows - Corrected Proof
L.V. Madoz, M.J. Giuliodori, M. Jaureguiberry, J. Plöntzke, M. Drillich, R.L. de la Sota

ABSTRACT: The objectives of this study were to assess the effect of the stage of estrous cycle on the percentage of endometrial polymorphonuclear cells (PMN) obtained by cytobrush to determine cutoff values for the diagnosis of subclinical endometritis under pastoral conditions, to measure the prevalence of subclinical endometritis 21 to 62 d in milk (DIM), and to evaluate the effect of subclinical endometritis on reproductive performance in grazing dairy cows. The first experiment was conducted on a commercial dairy farm in Buenos Aires province (Argentina), where 17 postpartum cyclic dairy cows without clinical endometritis were selected and synchronized by Ovsynch protocol. Endometrial cytology (cytobrush technique) and blood (tail vessels) samples were obtained on d 0, 4, 11, and 18 of the estrous cycle (corresponding to estrus, metestrus, diestrus, and proestrus, respectively) and used for measuring percentage of PMN and P4 concentration, respectively. The percentage of PMN was determined 3 times by blinded count by 2 operators. Data were analyzed with PROC MIXED, PROC GENMOD, and PROC FREQ from SAS 9.1. The percentage of PMN did not vary with the stage of the estrous cycle. In addition, PMN counts were below any of the reported thresholds in this study (4%) for most of the cows. Therefore, the risk for false positive test results as a consequence of physiological changes in the counts of PMN during estrous cycle is low. The second experiment was conducted on 4 commercial dairy farms in Buenos Aires province (Argentina), where lactating Holstein dairy cows (n = 418) 21 to 62 DIM without clinical endometritis were studied. Samples of endometrial cytology were collected with the cytobrush technique. Data were analyzed with receiver operator characteristic curves with Sigmaplot 10.0, and with PROC GLIMMIX, PROC PHREG, and PROC LIFETEST from SAS 9.1. Cutoff values for the diagnosis of subclinical endometritis in grazing dairy cows are 8% PMN for 21 to 33 DIM, 6% PMN for 34 to 47 DIM, 4% PMN for 48 to 62 DIM, and overall 5% PMN for 21 to 62 DIM; the prevalence of subclinical endometritis 21 to 62 DIM was 17%. Finally, subclinical endometritis diagnosed at 21 to 62 DIM decreases the hazard for pregnancy (hazard ratio = 0.668; 95% confidence interval = 0.492–0.909) and increases the calving to conception interval by d 30 compared with normal cows (median 95% confidence interval = 133 vs. 93, respectively).

Technical note: Identification of suitable normalizers for microRNA expression analysis in milk somatic cells of the yak (Bos grunniens) - Corrected Proof
W.L. Bai, R.H. Yin, R.J. Yang, W.A. Khan, Z.J. Ma, S.J. Zhao, W.Q. Jiang, Z.Y. Wang, Y.B. Zhu, G.B. Luo, Z.H. Zhao

ABSTRACT: MicroRNA are approximately 18- to 22-nucleotide nonprotein coding molecules that play important roles in the regulation of gene expression at the posttranscriptional level. In the present study, we assessed the suitability of 8 noncoding small RNA as normalizers for microRNA (miR) quantitative analysis in milk somatic cells of lactating yaks, including 3 small nuclear RNA (snRNA; RNU1A, RNU5A, and RNU6B), 3 small nucleolar RNA (snoRNA; SNORA73A, Z30, and SNORA74A), 1 rRNA (5S), and 1 transfer RNA (Met-tRNA). The snRNA RNU1A, RNU5A, and SNORA73A were identified as the most stable references in milk somatic cells of lactating yaks. Also, a minimum of 3 reference RNA (RNU1A, RNU5A, and SNORA73A) were required for the normalization of microRNA expression data in milk somatic cells of the lactating yak. We further evaluated the suitability of the combination of RNU1A, RNU5A, and SNORA73A as reference RNA in milk somatic cells of lactating yaks via detecting the relative expression of miR 16b, miR 21-5p, miR 145, and miR 155 as microRNA of putative interest. In comparison to the colostrum period, on the whole, the expressions of the 4 microRNA were found to be upregulated at an early period and, thereafter, a declining pattern was exhibited from early to final periods in all microRNA investigated. Based on the results from this study, we recommend that the combination of RNU1A, RNU5A, and SNORA73A can be used as normalizers for microRNA quantitative analysis in future longitudinal studies on milk somatic cells of lactating yaks in relation to lactation.

Technical note: A modified method to quantify prolamin proteins in dry and high-moisture corn - Corrected Proof
S.E. Nellis, P.C. Hoffman, R.D. Shaver

ABSTRACT: Numerous studies have evaluated laboratory methods to quantify prolamin proteins in dry corn; however, the utility of methods to quantify functional prolamins, which impede starch digestibility, in high-moisture corn (HMC) is less defined. As a result, a common rapid turbidimetric (rTM) laboratory procedure was modified (rapid Bradford method, rBM), extracting buffer-soluble proteins before prolamin solubilization in an effort to better quantify functional prolamins in HMC. Twenty samples of dry and HMC were evaluated by rTM and rBM procedures. Prolamin concentration in dry corn, as estimated by rTM or rBM methods, ranged from 6.12 to 2.20 g/100 g of DM or 5.46 to 2.41 g/100 g of DM, respectively. Dry corn mean prolamin concentrations, as estimated by rTM or rBM methods, were similar at 3.65 or 3.66 g/100 g of DM. Prolamin concentration in HMC, as estimated by rTM, ranged from 4.99 to 3.24 g/100 g of DM, with a mean prolamin concentration of 4.19 g/100 g of DM, but estimation of prolamins in HMC by the rBM method resulted in lower mean (4.19 vs. 3.24 g/100 g of DM) prolamin concentration. Prolamin concentration in dry and HMC measured by rTM was negatively related to peak absolute rates (PAR; mL/0.1 g of DM) of in vitro gas production. However, relationships between rTM prolamin concentration and PAR were not homogeneous and were different between dry and HMC. Prolamin proteins as determined by rBM were likewise negatively related to PAR, but corn type did not influence rBM prolamin concentration by PAR relationships. Data suggest that the rBM method defined more similar functional prolamin proteins, which impede starch degradability, in dry and HMC.

Effects of lasalocid and intermittent feeding of chlortetracycline on the growth of prepubertal dairy heifers - Corrected Proof
R.G. Cabral, P.S. Erickson, N.E. Guindon, E.J. Kent, C.E. Chapman, K.M. Aragona, M.D. Cabral, E.C. Massa, N.T. Antaya, C.C. Muir, B. O'Donnell, M.E. Branine

ABSTRACT: Forty Holstein heifers entered the 12-wk study at approximately 12 wk of age. At enrollment, heifers were blocked by birth date and assigned to 1 of 4 treatments: (1) carrier (30 g; control); (2) lasalocid + carrier (1 mg/kg of body weight; L); (3) chlortetracycline + carrier (22 mg/kg of body weight; CTC); (4) L + CTC + carrier (CTCL). Heifers on CTC and CTCL were provided treatment Monday through Friday and carrier only on Saturday and Sunday. These heifers were provided their respective treatment during wk 1 to 4, 6, and 10; wk 5, 7 to 9, and 11 to 12 heifers were provided the nonmedicated carrier. Heifers were individually fed a total mixed ration with treatments top-dressed at 1200 h daily. Dry matter intake was monitored for each heifer and feed provided was adjusted according to individual intakes. Skeletal measurements were taken weekly and blood samples were obtained every Monday, Wednesday, and Friday. Blood samples were analyzed for thyroxine concentration via radial immunoassay. Heifers supplemented with L had lower average daily gain, overall body weight gain, and trends for lower daily body length gain and overall girth gain compared with CTC heifers, but similar to control and CTCL heifers. Heifers fed L had lower hip height gain and overall hip height gain compared with CTCL heifers, but similar to control and CTC heifers. Heifers fed L had lower overall withers height gain compared with control heifers, but similar to CTC and CTCL heifers. No treatment effect on thyroxine concentrations was observed. These data indicate that L did not increase growth. Results from this experiment indicate that supplementing heifers with L was not beneficial and no benefits to supplementing heifers with CTC or the combination of CTC and L were evident compared with control heifers. Heifers in this study experienced minimal health problems and were regarded to be under low stress levels. Supplementing CTC and L may be beneficial to growing heifers under conditions where disease exposure and stressors are greater.

The amino acid composition of rumen-undegradable protein: A comparison between forages - Corrected Proof
B. Edmunds, K.-H. Südekum, R. Bennett, A. Schröder, H. Spiekers, F.J. Schwarz

ABSTRACT: The objective of this study was to improve knowledge regarding the amino acid profile of the insoluble portion of ingested forage escaping rumen degradation. Six forage categories were analyzed. Categories varied in botanical composition and each contained 2 samples. Samples within categories were derived from the same parent material but differed in harvest, maturity, or conservation type. The rumen-undegradable protein of all forages was measured by incubation for 16 h in the rumen of 3 nonlactating cows. All residues were corrected for microbial colonization. The AA profile of the residue was different to the original profile. Degradation trends of individual AA, in terms of increase or decrease relative to the original concentration, were similar between all forages. The AA profiles of forage residues, both within and between categories, were more similar to each other than to their respective original profile. This information may aid in improving the accuracy of estimating postruminal AA supply from forages while decreasing the number of samples required to be analyzed.

Short communication: Validation of a point-of-care glucometer for use in dairy cows - Corrected Proof
J.A M. Wittrock, T.F. Duffield, S.J. LeBlanc

ABSTRACT: The purpose of this study was to evaluate the diagnostic performance of a hand-held electronic glucometer (Precision Xtra; Abbott Diabetes Care Inc., Mississauga, ON, Canada) for cow-side use in dairy cattle. This device has been validated for measuring blood concentrations of β-hydroxybutyrate in dairy cows. This study was designed to assess the accuracy of whole-blood glucose measurements from the glucose meter relative to a reference chemical analyzer in a diagnostic laboratory. Duplicate samples were taken from the same cows at the same time, into blood tubes with either the glycolysis-inhibiting preservative sodium fluoride (NaF) or without preservative. Glucometer readings were taken on whole blood with no preservative, and laboratory measurements were conducted on serum preserved with NaF. Blood samples were collected from cows between 3 wk before and 5 wk after calving, including during a glucose tolerance test conducted 1 wk before expected calving. Passing-Bablok and Bland-Altman data analyses were used to evaluate the performance of the glucometer relative to the laboratory results. A strong correlation was observed in 709 samples from 81 cows between the hand-held meter and serum from samples preserved with NaF (R2 = 0.95). Overall, 96% of measurements with the glucometer fell within the 95% confidence limits of analysis in the laboratory, although at higher-than-physiologic glucose concentrations (>5.2 mmol/L) the glucometer tended to overestimate. The hand-held glucometer appears suitable for rapid measurement of glucose under field conditions in dairy cattle.

An automated walk-over weighing system as a tool for measuring liveweight change in lactating dairy cows - Corrected Proof
R.A. Dickinson, J.M. Morton, D.S. Beggs, G.A. Anderson, M.F. Pyman, P.D. Mansell, C.B. Blackwood

ABSTRACT: Automated walk-over weighing systems can be used to monitor liveweights of cattle. Minimal literature exists to describe agreement between automated and static scales, and no known studies describe repeatability when used for daily measurements of dairy cows. This study establishes the repeatability of an automated walk-over cattle-weighing system, and agreement with static electronic scales, when used in a commercial dairy herd to weigh lactating cows. Forty-six lactating dairy cows from a seasonal calving, pasture-based dairy herd in southwest Victoria, Australia, were weighed once using a set of static scales and repeatedly using an automated walk-over weighing system at the exit of a rotary dairy. Substantial agreement was observed between the automated and static scales when assessed using Lin's concordance correlation coefficient. Weights measured by the automated walkover scales were within 5% of those measured by the static scales in 96% of weighings. Bland and Altman's 95% limits of agreement were −23.3 to 43.6 kg, a range of 66.9 kg. The 95% repeatability coefficient for automated weighings was 46.3 kg. Removal of a single outlier from the data set increased Lin's concordance coefficient, narrowed Bland and Altman's 95% limits of agreement to a range of 32.5 kg, and reduced the 95% repeatability coefficient to 18.7 kg. Cow misbehavior during walk-over weighing accounted for many of the larger weight discrepancies. The automated walk-over weighing system showed substantial agreement with the static scales when assessed using Lin's concordance correlation coefficient. This contrasted with limited agreement when assessed using Bland and Altman's method, largely due to poor repeatability. This suggests the automated weighing system is inadequate for detecting small liveweight differences in individual cows based on comparisons of single weights. Misbehaviors and other factors can result in the recording of spurious values on walk-over scales. Excluding outlier weights and comparing means of 7 consecutive daily weights may improve agreement sufficiently to allow meaningful assessment of small short-term changes in automated weights in individuals and groups of cows.

Purification and identification of five novel antioxidant peptides from goat milk casein hydrolysates - Corrected Proof
Z. Li, A. Jiang, T. Yue, J. Wang, Y. Wang, J. Su

ABSTRACT: The present research described the preparation, purification, and identification of antioxidant peptides from goat milk casein (GMC). Goat milk casein was hydrolyzed by using a combination of neutral and alkaline proteases to obtain goat milk casein hydrolysates (GMCH) with high antioxidant activity. After desalting by nonpolar macroporous absorption resin, GMCH was isolated and purified by gel filtration chromatography and reversed-phase HPLC, respectively, and further identified by nanoliter electrospray ionization-tandem mass spectrometry. Antioxidant activities of GMC, GMCH, and pure peptides were evaluated and compared using free radical scavenging activity, metal ion chelating ability, and anti-lipid peroxidation ability. Compared with GMC, the free radical-scavenging ability and ferrous ion-chelating ability of GMCH increased significantly. The inhibition effect of lipid peroxidation of GMCH was much stronger than that of tert-butylhydroquinone and phytogermine and a little lower than that of ascorbic acid. The antioxidant activity of GMCH could be attributed to the high antioxidant activity of oligopeptides, especially 5 novel oligopeptides: Val-Tyr-Pro-Phe, Phe-Gly-Gly-Met-Ala-His, Phe-Pro-Tyr-Cys-Ala-Pro, Tyr-Val-Pro-Glu-Pro-Phe, and Tyr-Pro-Pro-Tyr-Glu-Thr-Tyr, which were first observed in GMCH. The antioxidant activity of these 5 novel oligopeptides and GMCH increased 3.59 to 380 times compared with GMC, combining anti-lipid peroxidation ability of GMCH, which indicated that GMCH and its purified fractions in different stages could be used as functional food ingredients, food additives, and pharmaceutical agents in the future.

Effect of castration method on neurohormonal and electroencephalographic stress indicators in Holstein calves of different ages - Corrected Proof
J.C. Dockweiler, J.F. Coetzee, L.N. Edwards-Callaway, N.M. Bello, H.D. Glynn, K.A. Allen, M.E. Theurer, M.L. Jones, K.A. Miller, L. Bergamasco

ABSTRACT: As public concern for food animal welfare increases, a need to validate objective pain assessment tools exists in order to formulate animal welfare policies and facilitate regulatory approval of compounds to alleviate pain in livestock in the United States. The aims of this study were (1) to compare the physiological response to pain induced by surgical and nonsurgical (band) castration in calves and (2) to elucidate age-related differences in pain response of calves subjected to different castration methods. Seventy-six Holstein bull calves were blocked by age (≤8-wk and ≥6-mo-old) and randomly assigned to 1 of 4 treatment groups: control (n = 20), castration by banding (n = 18), cut-and-clamp surgical castration (n = 20), and cut-and-pull surgical castration (n = 18). Measurements included electroencephalogram, heart rate variability, infrared thermography, electrodermal activity, and concentrations of serum cortisol and plasma substance P before, during, and within 20 min following castration. Electroencephalogram recordings showed desynchronization for all treatments, consistent with increased arousal; yet the magnitude of desynchronization was greatest for 6-mo-old calves castrated by cut-and-clamp. Additionally, older calves in the cut-and-pull group showed greater desynchronization than younger calves in the same group. Based on the heart rate variability analysis, 6-mo-old calves in the control or cut-and-pull castration groups showed greater sympathetic tone than younger calves in the same treatment groups. Overall, younger calves showed lower electrodermal activity than older calves. Regardless of treatment, concentrations of cortisol and plasma substance P were greater in 6-mo-old calves relative to their younger counterparts, indicating a more robust response to all treatments in older calves. In summary, neurohormonal and electroencephalographic stress responses of calves to castration were age-specific. Castration by cut-and-clamp showed the most pronounced stress response in 6-mo-old calves. These findings provide evidence that support welfare policies recommending castration at an early age and the use of analgesic compounds at the time of surgical castration especially in older calves. However, the potential long-term negative consequences of early untreated pain must be considered and warrant further investigation.

Comparison of different methods for imputing genome-wide marker genotypes in Swedish and Finnish Red Cattle - Corrected Proof
P. Ma, R.F. Brøndum, Q. Zhang, M.S. Lund, G. Su

ABSTRACT: This study investigated the imputation accuracy of different methods, considering both the minor allele frequency and relatedness between individuals in the reference and test data sets. Two data sets from the combined population of Swedish and Finnish Red Cattle were used to test the influence of these factors on the accuracy of imputation. Data set 1 consisted of 2,931 reference bulls and 971 test bulls, and was used for validation of imputation from 3,000 markers (3K) to 54,000 markers (54K). Data set 2 contained 341 bulls in the reference set and 117 in the test set, and was used for validation of imputation from 54K to high density [777,000 markers (777K)]. Both test sets were divided into 4 groups according to their relationship to the reference population. Five imputation methods (Beagle, IMPUTE2, findhap, AlphaImpute, and FImpute) were used in this study. Imputation accuracy was measured as the allele correct rate and correlation between imputed and true genotypes. Results demonstrated that the accuracy was lower when imputing from 3K to 54K than from 54K to 777K. Using various imputation methods, the allele correct rates varied from 93.5 to 97.1% when imputing from 3K to 54K, and from 97.1 to 99.3% when imputing from 54K to 777K; IMPUTE2 and Beagle resulted in higher accuracies and were more robust under various conditions than the other 3 methods when imputing from 3K to 54K. The accuracy of imputation using FImpute was similar to those results from Beagle and IMPUTE2 when imputing from 54K to high density, and higher than the remaining 2 methods. The results also showed that a closer relationship between test set and reference set led to a higher accuracy for all the methods. In addition, the correct rate was higher when the minor allele frequency was lower, whereas the correlation coefficient was lower when the minor allele frequency was lower. The results indicate that Beagle and IMPUTE2 provide the most robust and accurate imputation accuracies, but considering computing time and memory usage, FImpute is another alternative method.

Protein composition affects variation in coagulation properties of buffalo milk - Corrected Proof
V. Bonfatti, M. Gervaso, R. Rostellato, A. Coletta, P. Carnier

ABSTRACT: The aim of this study was to investigate the effects exerted by the content of casein and whey protein fractions on variation of pH, rennet-coagulation time (RCT), curd-firming time (K20), and curd firmness of Mediterranean buffalo individual milk. Measures of milk protein composition and assessment of genotypes at CSN1S1 and CSN3 were obtained by reversed-phase HPLC analysis of 621 individual milk samples. Increased content of αS1-casein (CN) was associated with delayed coagulation onset and increased K20, whereas average pH, RCT, and K20 decreased when β-CN content increased. Milk with low κ-CN content exhibited low pH and RCT relative to milk with high content of κ-CN. Increased content of glycosylated κ-CN was associated with unfavorable effects on RCT. Effects of milk protein composition on curd firmness were less important than those on pH, RCT, and K20. Likely, this occurred as a consequence of the very short RCT of buffalo milk, which guaranteed a complete strengthening of the curd even in the restricted 31 min time of analysis of coagulation properties and for samples initially showing soft curds. Effects of CSN1S1-CSN3 genotypes on coagulation properties were not to be entirely ascribed to existing variation in milk protein composition associated with polymorphisms at CSN1S1 and CSN3 genes. Although the role of detailed milk protein composition in variation of cheese yield needs to be further investigated, findings of this study suggest that modification of the relative content of specific CN fractions can relevantly influence the behavior of buffalo milk during processing.

Short communication: Aquaporin-7 mRNA in adipose depots of primiparous and pluriparous dairy cows: Long-term physiological and conjugated linoleic acid-induced changes - Corrected Proof
H. Sauerwein, B. Saremi, J. Pappritz, D. von Soosten, U. Meyer, S. Dänicke, M. Mielenz

ABSTRACT: Aquaglyceroporins act as channel proteins and regulate water and glycerol exchange through cell membranes. The aquaglyceroporin aquaporin-7 (AQP7) is abundantly expressed in adipose tissue (AT) and regulates the release of glycerol produced by lipolysis. We aimed to investigate the expression of AQP7 mRNA during lactation in subcutaneous (s.c.) and visceral (v.c.) adipose depots of primiparous and pluriparous dairy cows. In 2 independent experiments, Holstein cows were supplemented with conjugated linoleic acids (CLA) or a control (CON) fat supplement at 100 g/d. Pluriparous cows were supplemented starting with the first day in milk (DIM) up to 182 DIM and biopsies from s.c. AT were collected at d −21, 1, 21, 70, 105 182 196, 224, and 252 relative to calving (CLA = 11; CON = 10). Samples from 3 s.c. and v.c. adipose depots were investigated in primiparous cows (n = 25) receiving the supplements from 1 DIM until slaughter at 1, 42, or 105 DIM. The AQP7 mRNA abundance decreased from d −21 to 1 in s.c. AT of pluriparous cows without further increase to d 252 of lactation. In primiparous cows of the CON group, the AQP7 mRNA abundance increased from 1 to 105 DIM in s.c. AT from the tail head and in mesenteric AT. In retroperitoneal AT, the only depot for which a significant decrease in mass was observed with DIM, AQP7 mRNA abundance was greater at 42 and 105 than 1 DIM. Comparing the different fat depots, retroperitoneal AT had the highest and mesenterial AT had the lowest AQP7 mRNA abundance, but no general difference was observed between v.c. and s.c. fat depots. The values were not affected by CLA treatment with the exception of mesenteric AT, for which lower AQP7 mRNA abundance values were recorded in the CLA than in the CON group. The longitudinal characterization of the AQP7 mRNA expression profile throughout lactation revealed differences between primiparous and pluriparous cows, with an increase of AQP7 mRNA abundance up to 105 DIM only in the primiparous cows. Due to a lack of CLA effects in pluriparous cows and the limitation to just one fat depot in primiparous cows, a modulatory effect of CLA on AQP7 mRNA abundance in dairy cows is not supported by our study.

Effects of adjustable and stationary fans with misters on core body temperature and lying behavior of lactating dairy cows in a semiarid climate - Corrected Proof
S.D. Anderson, B.J. Bradford, J.P. Harner, C.B. Tucker, C.Y. Choi, J.D. Allen, L.W. Hall, S. Rungruang, R.J. Collier, J.F. Smith

ABSTRACT: Cows readily seek shade to reduce solar heat load during periods of high ambient temperature. Typically, auxiliary cooling systems are oriented to maximize cooling for shaded cows. However, when a shade structure is oriented north-south, stationary fan and mister cooling systems are unable to track shade as the sun's angle shifts throughout the day, and thus can become ineffective. The FlipFan Dairy Cooling System (Schaefer Ventilation Equipment, Sauk Rapids, MN) employs fans and misters that follow shade and compensate for wind speed by rotating on a horizontal axis. Multiparous, lactating Holstein cows (n = 144) on a commercial dairy in Arizona were cooled by a fixed system comprised of stationary fans and misters acting as control or the adjustable FlipFan operated for 16.5 h/d (0830 to 0100 h). Core body temperatures (CBT) of 64 cows (4 pens/treatment; 8 cows/pen; 6 d) and lying behavior of 144 cows (4 pens/treatment; 18 cows/pen; 5 d) were collected by intravaginal and leg data loggers, respectively. Cows were balanced by milk production, blocked by days in milk, and randomly assigned to pen within block. Pen was the experimental unit. In a second experiment, isothermal maps were developed using a fixed system of thermal data loggers arranged in the shaded areas of the pens at different times of day and were analyzed for differences in the temperature-humidity index (THI) achieved by each cooling treatment. Ambient conditions consisted of a mean temperature of 33.0°C, mean relative humidity of 40.3%, and mean THI of 80.2. Mean 24-h CBT for FlipFan was lower than control (38.9 vs. 39.1 ± 0.04°C). A treatment × time interaction was observed in which CBT of FlipFan was 0.4°C lower than control from 0600 to 0800 h and 1500 to 1600 h. Cows cooled by FlipFan spent more time lying down compared with those cooled by control (9.5 vs. 8.6 h/d). Cows under FlipFan had more frequent lying bouts than did those under control (12.8 vs. 10.7 bouts/d). Lower CBT and decreased standing time are consistent with the findings of other studies when ambient heat load was reduced. In the second experiment, the FlipFan system achieved a lower THI in the morning and evening (5.9 and 1.7%, respectively), and the THI also tended to be 0.8% lower in the afternoon compared with that of control. Results suggest that FlipFan is more effective than a stationary fan and mister system at decreasing CBT, increasing lying time and bouts, and providing a more desirable microenvironment for cows throughout the day in a semiarid environment.

Evaluation of inbreeding depression in Holstein cattle using whole-genome SNP markers and alternative measures of genomic inbreeding - Corrected Proof
D.W. Bjelland, K.A. Weigel, N. Vukasinovic, J.D. Nkrumah

ABSTRACT: The effects of increased pedigree inbreeding in dairy cattle populations have been well documented and result in a negative impact on profitability. Recent advances in genotyping technology have allowed researchers to move beyond pedigree analysis and study inbreeding at a molecular level. In this study, 5,853 animals were genotyped for 54,001 single nucleotide polymorphisms (SNP); 2,913 cows had phenotypic records including a single lactation for milk yield (from either lactation 1, 2, 3, or 4), reproductive performance, and linear type conformation. After removing SNP with poor call rates, low minor allele frequencies, and departure from Hardy-Weinberg equilibrium, 33,025 SNP remained for analyses. Three measures of genomic inbreeding were evaluated: percent homozygosity (FPH), inbreeding calculated from runs of homozygosity (FROH), and inbreeding derived from a genomic relationship matrix (FGRM). Average FPH was 60.5 ± 1.1%, average FROH was 3.8 ± 2.1%, and average FGRM was 20.8 ± 2.3%, where animals with larger values for each of the genomic inbreeding indices were considered more inbred. Decreases in total milk yield to 205 d postpartum of 53, 20, and 47 kg per 1% increase in FPH, FROH, and FGRM, respectively, were observed. Increases in days open per 1% increase in FPH (1.76 d), FROH (1.72 d), and FGRM (1.06 d) were also noted, as well as increases in maternal calving difficulty (0.09, 0.03, and 0.04 on a 5-point scale for FPH, FROH, and FGRM, respectively). Several linear type traits, such as strength (−0.40, −0.11, and −0.19), rear legs rear view (−0.35, −0.16, and −0.14), front teat placement (0.35, 0.25, 0.18), and teat length (−0.24, −0.14, and −0.13) were also affected by increases in FPH, FROH, and FGRM, respectively. Overall, increases in each measure of genomic inbreeding in this study were associated with negative effects on production and reproductive ability in dairy cows.

Microbiology of Cheddar cheese made with different fat contents using a Lactococcus lactis single-strain starter - Corrected Proof
J.R. Broadbent, C. Brighton, D.J. McMahon, N.Y. Farkye, M.E. Johnson, J.L. Steele

ABSTRACT: Flavor development in low-fat Cheddar cheese is typified by delayed or muted evolution of desirable flavor and aroma, and a propensity to acquire undesirable meaty-brothy or burnt-brothy off-flavor notes early in ripening. The biochemical basis for these flavor deficiencies is unclear, but flavor production in bacterial-ripened cheese is known to rely on microorganisms and enzymes present in the cheese matrix. Lipid removal fundamentally alters cheese composition, which can modify the cheese microenvironment in ways that may affect growth and enzymatic activity of starter or nonstarter lactic acid bacteria (NSLAB). Additionally, manufacture of low-fat cheeses often involves changes to processing protocols that may substantially alter cheese redox potential, salt-in-moisture content, acid content, water activity, or pH. However, the consequences of these changes on microbial ecology and metabolism remain obscure. The objective of this study was to investigate the influence of fat content on population dynamics of starter bacteria and NSLAB over 9 mo of aging. Duplicate vats of full fat, 50% reduced-fat, and low-fat (containing <6% fat) Cheddar cheeses were manufactured at 3 different locations with a single-strain Lactococcus lactis starter culture using standardized procedures. Cheeses were ripened at 8°C and sampled periodically for microbiological attributes. Microbiological counts indicated that initial populations of nonstarter bacteria were much lower in full-fat compared with low-fat cheeses made at all 3 sites, and starter viability also declined at a more rapid rate during ripening in full-fat compared with 50% reduced-fat and low-fat cheeses. Denaturing gradient gel electrophoresis of cheese bacteria showed that the NSLAB fraction of all cheeses was dominated by Lactobacillus curvatus, but a few other species of bacteria were sporadically detected. Thus, changes in fat level were correlated with populations of different bacteria, but did not appear to alter the predominant types of bacteria in the cheese.

Prednisolone and cefapirin act synergistically in resolving experimental Escherichia coli mastitis - Corrected Proof
Anja Sipka, Abhijit Gurjar, Suzanne Klaessig, Gerald E. Duhamel, Andrew Skidmore, Jantijn Swinkels, Peter Cox, Ynte Schukken

ABSTRACT: Mastitis in dairy cows is typically treated with intramammary antibiotics. The combination of antibiotics with corticosteroids tends to have a large market share where these products are registered. Our objective was to investigate the effect of prednisolone in combination with cefapirin on the inflammatory response of experimentally induced Escherichia coli mastitis. Six midlactating Holstein-Friesian cows were challenged in 3 quarters with E. coli and treated at 4, 12, 24, and 36 h postinfection with 300 mg of cefapirin in 1 quarter and a combination of 300 mg of cefapirin and 20 mg of prednisolone in another quarter. At 24 h (n = 3) or 48 h (n = 3) postinfection cows were euthanized for tissue sampling. Clinical scores, somatic cell count, and California mastitis test scores, as well as IL-1β, IFN-γ, IL-4, and IL-10 levels and bacterial growth in milk, were measured every 6 h. Experimental inoculation caused a moderate clinical mastitis in all cows in challenged, untreated quarters. The E. coli challenge strain was recovered from all infected quarters and confirmed by PCR-based fingerprinting. Challenged, untreated control quarters showed increased concentrations of all measured cytokines together with recruitment of polymorphonuclear neutrophilic leukocytes at 24 and 48 h postchallenge. Both treatments reduced udder swelling and sensitivity with no statistically significant difference between treatment groups. Administration of cefapirin alone or in combination with prednisolone resulted in significantly lower concentrations of IFN-γ, IL-1β, and IL-10 compared with challenged, untreated quarters. Treated quarters did show IL-4 production, but concentrations were significantly decreased compared with untreated, challenged quarters. Quarters treated with the combination of cefapirin and prednisolone showed a significantly lower concentration of IL-4 compared with cefapirin-only treatment. At both 24 and 48 h postinoculation, the level of polymorphonuclear neutrophilic leukocyte recruitment was lowest in challenged quarters treated with a combination of cefapirin and prednisolone, followed by cefapirin alone. Taken together, treatment with cefapirin alone inhibited bacterial growth in milk and reduced the host inflammatory responses. Addition of prednisolone to cefapirin had a synergistic effect, resulting in a lower density of leukocytes in tissue and milk and a quicker restoration of milk quality.

Nonstarter lactic acid bacteria volatilomes produced using cheese components - Corrected Proof
E. Sgarbi, C. Lazzi, G. Tabanelli, M. Gatti, E. Neviani, F. Gardini

ABSTRACT: In long-ripened cheese, flavor formation occurs during ripening. The metabolism of lactic acid bacteria (LAB) leads to the production of different compounds that contribute to the flavor of cheese. The contribution of LAB to the formation of cheese flavor has previously been studied. However, the specific nonstarter LAB (NSLAB) metabolic reactions in ripened cheese that lead to the formation of flavor compounds remain unclear. In ripened cheese, the nutrient sources available include small peptides or amino acids, citrate, lactate, free fatty acids, and starter LAB cell lysis products. Thus, the aim of this study was to evaluate the ability of NSLAB to produce volatile flavor compounds by using an in vitro system that used only the nutrients available in ripened cheese as the energy source. Moreover, the potential contribution of the NSLAB volatilome on total cheese flavor is discussed. For this purpose, the production of volatile compounds on cheese-based medium (CBM) and on starter LAB lysed cell medium (LCM) by 2 Lactobacillus casei and 2 Lactobacillus rhamnosus strains, previously isolated from ripened Parmigiano Reggiano cheese, was investigated. The generated volatile compounds were analyzed with head-space gas chromatography mass spectrometry. Overall, ketones, aldehydes, alcohols, and acids were the most abundant compounds produced. Differences in volatilome production were found between NSLAB grown in LCM and CBM. The catabolic metabolism of amino acids and fatty acids were required for NSLAB growth on LCM. Conversely, pyruvate metabolism was the main catabolic pathway that supported growth of NSLAB in CBM. This study can be considered a first step toward a better understanding of how microbiota involved in the long ripening of cheese may contribute to the development of cheese flavor.

Replacing alfalfa silage with corn silage in dairy cow diets: Effects on enteric methane production, ruminal fermentation, digestion, N balance, and milk production - Corrected Proof
F. Hassanat, R. Gervais, C. Julien, D.I. Massé, A. Lettat, P.Y. Chouinard, H.V. Petit, C. Benchaar

ABSTRACT: The objective of this study was to determine the effects of replacing alfalfa silage (AS) with corn silage (CS) in dairy cow total mixed rations (TMR) on enteric CH4 emissions, ruminal fermentation characteristics, apparent total-tract digestibility, N balance, and milk production. Nine ruminally cannulated lactating cows were used in a replicated 3 × 3 Latin square design (32-d period) and fed (ad libitum) a TMR [forage:concentrate ratio of 60:40; dry matter (DM) basis], with the forage portion consisting of either alfalfa silage (0% CS; 56.4% AS in the TMR), a 50:50 mixture of both silages (50% CS; 28.2% AS and 28.2% CS in the TMR), or corn silage (100% CS; 56.4% CS in the TMR). Increasing the CS proportion (i.e., at the expense of AS) in the diet was achieved by decreasing the corn grain proportion and increasing that of soybean meal. Intake of DM and milk yield increased quadratically, whereas DM digestibility increased linearly as the proportion of CS increased in the diet. Increasing the dietary CS proportion resulted in changes (i.e., lower ruminal pH and acetate:propionate ratio, reduced fiber digestibility, decreased protozoa numbers, and lower milk fat and higher milk protein contents) typical of those observed when cows are fed high-starch diets. A quadratic response in daily CH4 emissions was observed in response to increasing the proportion of CS in the diet (440, 483, and 434 g/d for 0% CS, 50% CS, and 100% CS, respectively). Methane production adjusted for intake of DM, and gross or digestible energy was unaffected in cows fed the 50% CS diet, but decreased in cows fed the 100% CS diet (i.e., quadratic effect). Increasing the CS proportion in the diet at the expense of AS improved N utilization, as reflected by the decreases in ruminal NH3 concentration and manure N excretion, suggesting low potential NH3 and N2O emissions. Results from this study, suggest that total replacement of AS with CS in dairy cow diets offers a means of decreasing CH4 output and N losses. However, the reduction in fiber degradation and the resulting increase in volatile solids content of the manure may lead to increased CH4 emissions from manure storage.

Elimination kinetics of cephapirin sodium in milk after an 8-day extended therapy program of daily intramammary infusion in healthy lactating Holstein-Friesian cows - Corrected Proof
P.J. Gorden, M. van der List, F.D. Lehman, R.K. Lantz, P.D. Constable

ABSTRACT: The objective of this study was to determine the elimination kinetics of extended therapy with intramammary (IMM) cephapirin in lactating dairy cattle. Eight healthy Holstein-Friesian cows were administered cephapirin (200 mg) into all 4 mammary glands every 24 h after milking. Cows were milked 3 times per day and concentrations of cephapirin and desacetyl cephapirin were determined in bucket milk using liquid chromatography-mass spectrometry. Milk concentration-time data after the last of the 8 IMM infusions were fitted using compartment and noncompartmental models. The maximum cephapirin concentration was 128 ± 57 μg/mL (mean ± SD), the elimination rate constant from the central compartment was 0.278 ± 0.046 (h−1), clearance was 0.053 ± 0.023 L/h, the half time for elimination was 2.55 ± 0.40 h, and the mean residence time was 2.65 ± 0.79 h. The cephapirin concentration was below the approved tolerance in all cows by 96 h after the last infusion, which is the labeled withholding time for the preparation used. Extended therapy for 8 d provided milk cephapirin concentrations above the minimum inhibitory concentration for common gram-positive mastitis pathogens (0.1 to 1.0 μg/mL) for the duration of therapy and for an additional 16 to 32 h after the end of treatment. Our findings suggest that this IMM cephapirin sodium formulation, which is labeled for 2 doses 12 h apart, could be administered at a 24-h interval for up to 8 d in cows milked 3 times per day, with no significant effect on residue levels by 96 h after the last treatment. Longer withdrawal times would be prudent for cows with low milk production.

Behavioral and physiological effects of a short-term feed restriction in lactating dairy cattle with different body condition scores at calving - Corrected Proof
K.E. Schütz, N.R. Cox, K.A. Macdonald, J.R. Roche, G.A. Verkerk, A.R. Rogers, C.B. Tucker, L.R. Matthews, S. Meier, J.R. Webster

ABSTRACT: Body condition score (BCS) around calving, and the typical BCS loss for up to 100 d after parturition, is associated with both production and reproductive performance of dairy cattle. In addition, there is public concern that thin cows may have impaired welfare, particularly in early lactation where feed demand exceeds pasture growth, and a lag exists between peak milk energy requirements and intake. The aim of this experiment was to determine how BCS at calving influences behavioral and physiological responses to a short-term feed restriction at 47 DIM. Body condition score (on a 10-point scale) at calving was manipulated by modifying the diets in the previous lactation of healthy dairy cattle to generate 3 treatment groups: low BCS (3.4; n = 17), medium BCS (4.6; n = 18), or high BCS (5.4; n = 20). Cows were tested in 4 groups for 8 consecutive days; testing consisted of different levels of feed allocation (d 1 and 2: 100%; d 3 and 4: 75%; d 5: 50%; d 6 to 8: 125%), where 100% was 15 kg of DM/cow per day. All BCS groups had similar and marked behavioral and physiological responses to feed restriction. For example, they increased vocalization, time spent eating silage and grazing, aggressive behavior, and fat metabolism (as measured by concentrations of β–hydroxybutyrate and nonesterified fatty acids), and reduced milk production. Body condition affected some of these responses. Fewer cows with low BCS engaged in aggressive interactions in a feed competition test (trough filled with silage that could be consumed in 15 min) on the first day of feed restriction (low: 32%; medium: 74%; high: 64%; standard error of difference = 15.4%). High-BCS cows had greater concentrations of β–hydroxybutyrate and nonesterified fatty acids throughout the experimental period, which suggests more fat mobilization; however, plasma leptin and fecal glucocorticosteroid metabolite concentrations were unaffected by BCS. Whereas cows demonstrated marked responses to feed restriction, the results suggest that a BCS of 3.4, 4.6, or 5.4 in healthy cows at calving does not overwhelmingly influence this response at 47 DIM.

Effects of gas composition in headspace and bicarbonate concentrations in media on gas and methane production, degradability, and rumen fermentation using in vitro gas production techniques - Corrected Proof
Amlan Kumar Patra, Zhongtang Yu

ABSTRACT: Headspace gas composition and bicarbonate concentrations in media can affect methane production and other characteristics of rumen fermentation in in vitro gas production systems, but these 2 important factors have not been evaluated systematically. In this study, these 2 factors were investigated with respect to gas and methane production, in vitro digestibility of feed substrate, and volatile fatty acid (VFA) profile using in vitro gas production techniques. Three headspace gas compositions (N2 + CO2 + H2 in the ratio of 90:5:5, CO2, and N2) with 2 substrate types (alfalfa hay only, and alfalfa hay and a concentrate mixture in a 50:50 ratio) in a 3 × 2 factorial design (experiment 1) and 3 headspace compositions (N2, N2 + CO2 in a 50:50 ratio, and CO2) with 3 bicarbonate concentrations (80, 100, and 120 mM) in a 3 × 3 factorial design (experiment 2) were evaluated. In experiment 1, total gas production (TGP) and net gas production (NGP) was the lowest for CO2, followed by N2, and then the gas mixture. Methane concentration in headspace gas after fermentation was greater for CO2 than for N2 and the gas mixture, whereas total methane production (TMP) and net methane production (NMP) were the greatest for CO2, followed by the gas mixture, and then N2. Headspace composition did not affect in vitro digestibility or the VFA profile, except molar percentages of propionate, which were greater for CO2 and N2 than for the gas mixture. Methane concentration in headspace gas, TGP, and NGP were affected by the interaction of headspace gas composition and substrate type. In experiment 2, increasing concentrations of CO2 in the headspace decreased TGP and NGP quadratically, but increased the concentrations of methane, NMP, and in vitro fiber digestibility linearly, and TMP quadratically. Fiber digestibility, TGP, and NGP increased linearly with increasing bicarbonate concentrations in the medium. Concentrations of methane and NMP were unaffected by bicarbonate concentration, but TMP tended to increase due to increasing bicarbonate concentration. Although total VFA concentration and molar percentage of butyrate were unchanged, the molar percentage of acetate, and acetate-to-propionate ratio decreased, whereas the molar percentage of propionate increased quadratically with increasing bicarbonate concentration. This study demonstrated for the first time that headspace composition, especially CO2 content, and bicarbonate concentration in media could significantly influence gas and methane production, and rumen fermentation in gas production techniques.

Particle size alterations of feedstuffs during in situ neutral detergent fiber incubation - Corrected Proof
M. Krämer, P. Nørgaard, P. Lund, M.R. Weisbjerg

ABSTRACT: Particle size alterations during neutral detergent fiber (NDF) determination and in situ rumen incubation were analyzed by dry sieving and image analysis to evaluate the in situ procedure for estimation of NDF degradation parameters and indigestible NDF concentration in terms of particle size. Early-cut and late-cut grass silages, corn silage, alfalfa silage, rapeseed meal, and dried distillers grains were examined. Treatments were (1) drying and grinding of forage samples and grinding of concentrates; (2) neutral detergent-soluble (NDS) extraction; (3) machine washing and NDS extraction; (4) 24-h rumen incubation, machine washing, and NDS extraction; and (5) 288-h rumen incubation, machine washing, and NDS extraction. Degradation profiles for potentially degradable NDF were determined and image analysis was used to estimate particle size profiles and thereby the risk for particle loss. Particle dimensions changed during NDF determination and in situ rumen incubation and variations depended on feedstuff and treatment. Corn silage and late-cut grass silage varied most in particle area among feedstuffs, with an increase of 139% between 0 and 24 h and a decrease of 77% between 24 and 288 h for corn silage and a decrease of 74% for late-cut grass silage between 24- and 288-h in situ rumen incubation. Especially for late-cut grass silage residues after 288 h in situ rumen incubation, a high mass proportion in the critical zone for escape was found. Particle area decreased linearly with increasing incubation time. Particle loss during in situ rumen incubation cannot be excluded and is likely to vary among feedstuffs.

Hypophagic effects of propionic acid are not attenuated during a 3-day infusion in the early postpartum period in Holstein cows - Corrected Proof
S.E. Stocks, M.S. Allen

ABSTRACT: We previously showed that propionic acid was more hypophagic than acetic acid when infused intraruminally in cows in the postpartum period and that the degree of hypophagia from short-term propionic acid infusion (18 h) was related to the acetyl coenzyme A (CoA) concentration in the liver. The objective of this experiment was to evaluate adaptation over time with longer-term infusions over 3 d. Twelve multiparous cows (2–13 d postpartum) were blocked by calving date and assigned randomly to treatment sequence in a crossover design experiment. The experiment was 12 d long with covariate periods preceding each 3-d infusion period. Treatments were 1.0 M propionic acid or 1.0 M acetic acid, infused intraruminally at 0.5 mol of volatile fatty acids/h beginning 6 h before feeding and continuing for 78 h with 3 d between infusions. Propionic acid decreased dry matter intake (DMI) relative to acetic acid (15.9 vs. 17.0 kg/d). However, a period-by- treatment interaction was detected for DMI. During period 1, propionic acid decreased DMI relative to acetic acid (14.3 vs. 17.5 kg/d) because of a reduction in meal size (1.30 vs. 1.65 kg), with no effect on intermeal interval. Propionic acid decreased DMI over the first 4 h following feeding (5.86 vs. 8.23 kg) but did not affect DMI 4 to 24 h after feeding. The depression in DMI in period 1 was positively related to hepatic acetyl-CoA concentration during the covariate period. Propionic acid was increasingly more hypophagic than acetic acid as hepatic acetyl-CoA concentration was elevated. No treatment-by-day interaction for DMI was observed, suggesting little or no measurable adaptation to treatment over the 3-d infusion period. These results suggest that hypophagia from propionic acid is enhanced when hepatic acetyl-CoA concentrations are elevated, such as when cows are in a lipolytic state.

Effect of early exposure to different feed presentations on feed sorting of dairy calves - Corrected Proof
E.K. Miller-Cushon, R. Bergeron, K.E. Leslie, G.J. Mason, T.J. DeVries

ABSTRACT: This study examined how early exposure to different feed presentations affects development of feed sorting in dairy calves. Twenty Holstein bull calves were exposed for the first 8 wk of life to 1 of 2 feed presentation treatments: concentrate and chopped grass hay (<2.5 cm) offered ad libitum at a ratio of 7:3 as a mixture (MIX), or as separate components (COM). Calves received 8 L/d of milk replacer (1.2 kg of dry matter), with the amount progressively reduced after 5 wk to facilitate weaning by the end of wk 7. All calves received the MIX diet in wk 9 to 11 and, subsequently, a novel total mixed ration (TMR; containing 40.5% corn silage, 22.0% haylage, 21.5% high-moisture corn, and 16.0% protein supplement) in wk 12 to 13. Intake was recorded daily and calves were weighed twice a week. Fresh feed and orts were sampled on d 1 to 4 of wk 6, 8, 9, 11, 12, and 13 for analysis of feed sorting, which was assessed through nutrient analysis for the MIX diet and particle size analysis for the TMR. The particle separator had 3 screens (19, 8, and 1.18 mm) producing long, medium, short, and fine particle fractions. Sorting of nutrients or particle fractions was calculated as the actual intake as a percentage of predicted intake; values >100% indicate sorting for, whereas values <100% indicate sorting against. Feed presentation did not affect dry matter intake or growth. Prior to weaning, all calves selected in favor of hay; MIX calves consumed more neutral detergent fiber (NDF) than predicted (103.6%) and less nonfiber carbohydrates (NFC) than predicted (92.6%), and COM calves consumed, as a percentage of dry matter intake, 40.3% hay (vs. 30% offered rate). In wk 8, calves fed COM consumed more NFC than calves fed MIX (1.0 vs. 0.95 kg/d) and less NDF (0.43 vs. 0.54 kg/d), indicating greater selection in favor of concentrate. However, when provided the MIX diet, calves previously fed COM did not sort, whereas calves previously fed MIX consumed more NFC intake than predicted (103.2%) and less NDF intake than predicted (97.6%). Calves previously fed MIX maintained increased sorting after transition to the novel TMR, sorting against long particles (86.5%) and for short (101.8%) and fine (101.2%) particles. These results indicate that initially providing dairy calves with solid feeds as separate components, compared with as a mixed ration, reduces the extent of feed sorting in the weeks after transition to a common ration.

β-Lactoglobulin-linoleate complexes: In vitro digestion and the role of protein in fatty acids uptake - Corrected Proof
Solène Le Maux, André Brodkorb, Thomas Croguennec, Alan A. Hennessy, Saïd Bouhallab, Linda Giblin

ABSTRACT: The dairy protein β-lactoglobulin (BLG) is known to bind fatty acids such as the salt of the essential long-chain fatty acid linoleic acid (cis, cis-9,12-octadecadienoic acid, n-6, 18:2). The aim of the current study was to investigate how bovine BLG-linoleate complexes, of various stoichiometry, affect the enzymatic digestion of BLG and the intracellular transport of linoleate into enterocyte-like monolayers. Duodenal and gastric digestions of the complexes indicated that BLG was hydrolyzed more rapidly when complexed with linoleate. Digested as well as undigested BLG-linoleate complexes reduced intracellular linoleate transport as compared with free linoleate. To investigate whether enteroendocrine cells perceive linoleate differently when part of a complex, the ability of linoleate to increase production or secretion of the enteroendocrine satiety hormone, cholecystokinin, was measured. Cholecystokinin mRNA levels were different when linoleate was presented to the cells alone or as part of a protein complex. In conclusion, understanding interactions between linoleate and BLG could help to formulate foods with targeted fatty acid bioaccessibility and, therefore, aid in the development of food matrices with optimal bioactive efficacy.

Use of dairies by postreproductive flocks of European starlings - Corrected Proof
H.J. Homan, J.T. LeJeune, D.L. Pearl, T.W. Seamans, A.A. Slowik, Mark R. Morasch, G.M. Linz

ABSTRACT: Knowledge of the behavior and movement patterns of European starlings (Sturnus vulgaris L.) is important to wildlife managers that seek to resolve conflicts at livestock facilities. We captured and radio tagged 10 starlings at each of 5 dairies in northeastern Ohio. From September 19 to October 31, 2007, we obtained sufficient data from 40 birds to study their behavior and movements. The birds visited the dairies where they were initially captured (home sites) on 85% of the days, spending 58% of each day at the dairies. Onsite arrival and departure times were 2.5 h after sunrise and 3.1 h before sunset. Daily visits by radio-tagged cohorts from the other dairies were greatest for the 2 most proximate dairies (1.3 km apart), with number of visits between this pairing >7× that of the 9 other pairings combined (4.1–6.5 km apart). Two birds used their home sites intermittently as roosts, arriving 3.8 h before sunset and departing 0.2 h after sunrise. In addition to using home-site roosts, these birds also used a distant roost (22 km) that was used by 36 of the 40 birds. The efficacy of starling management programs, especially lethal management, depends on degree of site fidelity, use of other facilities, and roosting behavior. For example, starlings that use dairies as roosting sites may require a different management strategy than required at dairies used as daytime sites because of differences in arrival and departure behavior. Our research will help resource managers evaluate current management strategies already in place and change them, if needed, to fit the behavior profile of starlings using dairies and other types of livestock facilities.

Comparison of 2 systems of pasture allocation on milking intervals and total daily milk yield of dairy cows in a pasture-based automatic milking system - Corrected Proof
N.A. Lyons, K.L. Kerrisk, S.C. Garcia

ABSTRACT: Cows milked in pasture-based automatic milking systems (AMS) have greater milking intervals than cows milked in indoor AMS. Long milking intervals greater than 16 h have a negative effect on milk yield and udder health. The impact of 2 systems of pasture allocation in AMS on milking interval and yield was investigated at the FutureDairy AMS research farm (Elizabeth Macarthur Agricultural Institute, New South Wales Department of Primary Industries, Camden, New South Wales, Australia) in late November to early December 2010. Two- (2WG) versus 3-way grazing (3WG) allocations per 24-h period were compared in a field study to test the hypothesis that an increase in the frequency of pasture allocation would reduce the milking interval and, therefore, increase milking frequency. The study involved the entire milking herd of 145 cows, with (mean ± SD) DIM = 121 ± 90 d, 7-d average milking frequency = 1.52 ± 0.41 milkings/cow per day, and 7-d average milk yield = 21.3 ± 7.6 kg/cow per day. Cows were milked using 2 DeLaval VMS milking units (DeLaval International AB, Tumba, Sweden). Cows in the 3WG treatment had 31% reduced milking interval, 40% greater milking frequency, and 20% greater daily milk production compared with 2WG. Increased milking frequency and milk production for 3WG was associated with greater utililization levels of the AMS milking units throughout the day. These results support the recommendation that, wherever possible, farmers installing AMS should incorporate sufficient infrastructure to accommodate 3WG, which provides additional flexibility with managing extremely long (and short) milking intervals.

Estimate of serum immunoglobulin G concentration using refractometry with or without caprylic acid fractionation - Corrected Proof
K.M. Morrill, J. Polo, A. Lago, J. Campbell, J. Quigley, H. Tyler

ABSTRACT: Objectives of this study were to develop a rapid calf-side test to determine serum IgG concentrations using caprylic acid (CA) fractionation, followed by refractometry of the IgG-rich supernatant and compare the accuracy of this method with results obtained using refractometry using raw serum. Serum samples (n = 200) were obtained from 1-d-old calves, frozen (−20°C), and shipped to the laboratory. Samples were allowed to thaw for 1 h at room temperature. Fractionation with CA was conducted by adding 1 mL of serum to a tube containing 45, 60, or 75 μL of CA and 0.5, 1.0, or 1.5 mL of 0.06 M acetic acid. The tube contents were mixed well, allowed to react for 1 min, and then centrifuged at 3,300 × g for 0, 10, or 20 min at 25°C. The %Brix and refractive index of the fractionated supernatant were determined using a digital refractometer. Nonfractionated serum was analyzed for %Brix (BRn), refractive index (nDn), and IgG concentration by radial immunodiffusion. The mean serum IgG concentration was 19.0 mg/mL [standard deviation (SD) = 9.7], with a range of 3.5 to 47.0 mg/mL. The mean serum BRn was 8.6 (SD = 0.91), with a range of 6.8 to 11.0. The mean serum nDn was 1.34566 (SD = 0.00140), with a range of 1.34300 to 1.34930. Serum nDn was positively correlated with IgG concentration (correlation coefficient = 0.86; n = 185). Fractionated samples treated with 1 mL 0.6 M acetic acid and 60 μL of CA and not centrifuged before analysis resulted in a strong relationship between the refractive index of the fractionated supernatant and IgG (correlation coefficient = 0.80; n = 45). Regression was used to determine cut points indicative of 10, 12, and 14 mg of IgG/mL to determine the sensitivity and specificity of refractometry to identify failure of passive transfer (serum IgG <10 mg/mL at 24 h old). The nDn were 1.34414, 1.34448, and 1.34480 to predict 10, 12, and 14 mg of IgG/mL of serum, respectively. The BRn cut points were 7.6, 7.8, and 8.0, respectively. The nDn cut points of 1.34448 and 1.34480 resulted in similar specificities (82.9%), whereas the 1.34414 cut point had a specificity of 60.0%. The BRn cut point of 7.6 and 7.8%Brix resulted in a similar percentage of correctly classified samples (89.7 and 90.8%, respectively); however, the 7.8% Brix cut point resulted in fewer false positives. These results suggest that Brix refractometry of nonfractionated calf serum provides a strong estimate of IgG concentration and 7.8% Brix may be used as the cut point to identify failure of passive transfer in 1-d-old calves.

Storage of refrigerated raw goat milk affecting the quality of whole milk powder - Corrected Proof
C.R. Fonseca, K. Bordin, A.M. Fernandes, C.E.C. Rodrigues, C.H. Corassin, A.G. Cruz, C.A.F. Oliveira

ABSTRACT: The aim of this study was to evaluate the influence of the growth of lipolytic bacteria in raw goat milk stored under refrigeration for different periods on quality parameters of goat milk powder during its shelf life. Fresh goat milk (100 L) was collected after milking, divided into 3 identical fractions, and stored at 4°C for 1, 3, and 5 d. On d 1, 3, and 5, one sample (1 L) was collected and used for microbiological and chemical analysis, and the remaining fraction (almost 30 L) was spray dried and stored at 25°C. Milk powder was submitted to microbiological, chemical, and sensory analysis immediately after production, and on d 60, 120, and 180. Lipolytic psychrotrophic counts and total free fatty acid content did not increase in raw milk during storage. However, peroxide value, caprylic and capric acid concentrations, and total free fatty acid content of milk powder increased during 180 d of storage, with higher levels found in milk powder manufactured with raw milk stored for 5 d. Capric odor and rancid flavors increased in milk powder during storage, regardless the of storage of raw milk for 1, 3, or 5 d. Heat treatments used during powder processing destroyed lipolytic psychrotrophic bacteria, but did not prevent lipolysis in milk powder. Results of this trial indicate that the storage of raw goat milk at 4°C should not exceed 3 d to preserve the quality of goat milk powder during its shelf life of 180 d.

Reaction norm of fertility traits adjusted for protein and fat production level across lactations in Holstein cattle - Corrected Proof
A. Menendez-Buxadera, M.J. Carabaño, O. Gonzalez-Recio, R.I. Cue, E. Ugarte, R. Alenda

ABSTRACT: A total of 304,001 artificial insemination outcomes in up to 7 lactations from 142,389 Holstein cows, daughters of 5,349 sires and 101,433 dams, calving between January 1995 and December 2007 in 1,347 herds were studied by a reaction norm model. The (co)variance components for days to first service (DFS), days open, nonreturn rate in the first service (NRFS), and number of services per conception were estimated by 6 models: 3 Legendre polynomial degrees for the genetic effects and adjustment or not for the level of fat plus protein (FP) production recorded at day closest to DFS. For all traits and type of FP adjustment, a second degree polynomial showed the best fit. The use of the adjusted FP model did not increase the level of genetic (co)variance components except for DFS. The heritability for each of the traits was low in general (0.03–0.10) and increased from the first to fourth calving, nevertheless a very important variability was found for the estimated breeding value (EBV) of the sires. The genetic correlations (rg) were close to unity between adjacent calvings, but decreased for most distant parities, ranging from rg = 0.36 (for DFS) to rg = 0.63 (for NRFS), confirming the existence of heterogeneous genetic (co)variance components and EBV across lactations. The results of the eigendecomposition of rg shows that the first eigenvalue explained between 82 to 92% and the second between 8 to 14% of the genetic variance for all traits; therefore, a deformation of the overall mean trajectory for reproductive performance across the trajectory of the different calving could be expected if selection favored these eigenfunctions. The results of EBV for the 50 best sires showed a substantial reranking and variation in the shape of response across lactations. The more important aspect to highlight, however, is the difference between the EBV of the same sires in different calvings, a characteristic known as plasticity, which is particularly important for DFS and NRFS. This component of fertility adds another dimension to selection for fertility that can be used to change the negative genetic progress of reproductive performance presented in this population of Holstein cows. The use of a reaction norm model should allow producers to obtain more robust cows for maintenance of fertility levels along the whole productive life of the cows.

Prediction of urinary nitrogen and urinary urea nitrogen excretion by lactating dairy cattle in Northwestern Europe and North America: A meta-analysis - Corrected Proof
J.W. Spek, J. Dijkstra, G. van Duinkerken, W.H. Hendriks, A. Bannink

ABSTRACT: A meta-analysis was conducted on the effect of dietary and animal factors on the excretion of total urinary nitrogen (UN) and urinary urea nitrogen (UUN) in lactating dairy cattle in North America (NA) and northwestern Europe (EU). Mean treatment data were used from 47 trials carried out in NA and EU. Mixed model analysis was used with experiment included as a random effect and all other factors, consisting of dietary and animal characteristics, included as fixed effects. Fixed factors were nested within continent (EU or NA). A distinction was made between urinary excretions based on either urine spot samples or calculated assuming a zero N balance, and excretions that were determined by total collection of urine only. Moreover, with the subset of data based on total collection of urine, a new data set was created by calculating urinary N excretion assuming a zero N balance. Comparison with the original subset of data allowed for examining the effect of such an assumption on the relationship established between milk urea N (MUN) concentration and UN. Of all single dietary and animal factors evaluated to predict N excretion in urine, MUN and dietary crude protein (CP) concentration were by far the best predictors. Urinary N excretion was best predicted by the combination of MUN, CP, and dry matter intake, whereas UUN was best predicted by the combination of MUN and CP. All other factors did not improve or only marginally improved the prediction of UN or UUN. The relationship between UN and MUN differed between NA and EU, with higher estimated regression coefficients for MUN for the NA data set. Precision of UN and UUN prediction improved substantially when only UN or UUN data based on total collection of urine were used. The relationship between UN and MUN for the NA data set, but not for the EU data set, was substantially altered when UN was calculated assuming a zero N balance instead of being based on the total collection of urine. According to results of the present meta-analysis, UN and UUN are best predicted by the combination of MUN and CP and that, in regard to precision and accuracy, prediction equations for UN and UUN should be derived from the total collection of urine.

Invited review: Heat stress effects during late gestation on dry cows and their calves - Corrected Proof
S. Tao, G.E. Dahl

ABSTRACT: In dairy cattle, late gestation is a critical period for fetal growth and physiological transition into the next lactation. Environmental factors, such as temperature and light, exert dramatic effects on the production, health, and well-being of animals during this period and after parturition. The aim of this review was to introduce effects of heat stress during late gestation on dairy cattle, and discuss the biological mechanisms that underlie the observed production and health responses in the dam and her fetus. Relative to cooled cows, cows that are heat stressed during late gestation have impaired mammary growth before parturition and decreased milk production in the subsequent lactation. In response to higher milk yield, cows cooled prepartum undergo a series of homeorhetic adaptations in early lactation to meet higher demand for milk synthesis compared with heat-stressed cows, but no direct effect of environmental heat stress on metabolism exists during the dry period. Prepartum cooling improves immune status of transition cows and evidence suggests that altered prolactin signaling in immune cells mediates the effects of heat stress on immune function. Late-gestation heat stress compromises placental development, which results in fetal hypoxia, malnutrition, and eventually fetal growth retardation. Maternal heat stress may also have carryover effects on the postnatal growth of offspring, but direct evidence is still lacking. Emerging evidence suggests that offspring from prepartum heat-stressed cows have compromised passive immunity and impaired cell-mediated immune function compared with those from cooled cows.

Descriptive analysis and early-stage consumer acceptance of yogurts fermented with carrot juice - Corrected Proof
M.A. Cliff, L. Fan, K. Sanford, K. Stanich, C. Doucette, N. Raymond

ABSTRACT: This research explored the sensory characteristics and consumer acceptance of novel probiotic unsweetened yogurts. Yogurts were made with 4 carrot juice levels (8, 16, 24, and 32%), 2 firmness levels (regular, 45 g/L milk solids; firm, 90 g/L milk solids), and 2 starter cultures (C1, C2). The sensory profile characterized the color intensity (before and after stirring), carrot flavor, sourness, and 7 texture/mouth-feel attributes (astringency, chalkiness, mouth-coating, thickness, smoothness, creaminess, and graininess). The influence of carrot juice level and firmness level were evaluated using ANOVA, polynomial contrasts, and principal component analysis. Mean scores and standard errors were calculated. Consumer acceptance panels in Wolfville, Nova Scotia (n = 56), and in Vancouver, British Columbia (Asian n = 72, non-Asian n = 72), evaluated the hedonic responses to the C1 and C2 formulations, respectively. We observed increases in color intensity, carrot flavor, creaminess, mouth-coating, and chalkiness with increasing carrot juice levels, as well as increases in color intensity, carrot flavor, creaminess, mouth-coating, thickness, and astringency with increasing milk solids concentrations of the C1 and C2 yogurts. Mean hedonic scores for color, appearance, and texture/mouth-feel were greater than hedonic scores for aroma, flavor/taste, and overall liking. This research identified the sensory qualities that need further development and demonstrated the importance of early-stage consumer acceptance research for directing new product development.

Concentrations of n-3 and n-6 fatty acids in Dutch bovine milk fat and their contribution to human dietary intake - Corrected Proof
H.J.F. van Valenberg, K.A. Hettinga, J. Dijkstra, H. Bovenhuis, E.J.M. Feskens

ABSTRACT: Weekly samples representative of Dutch milk were analyzed for concentrations of n-3 and n-6 fatty acids (FA). Concentrations of the n-3 FA α-linolenic acid (ALA), eicosatetraenoic acid, eicosapentaenoic acid, and docosapentaenoic acid were 0.495 ± 0.027, 0.041 ± 0.004, 0.067 ± 0.005, and 0.086 ± 0.008 g per 100 g of fat, respectively, whereas docosahexaenoic acid was absent or present in concentrations lower than 0.020 g per 100 g of fat. Concentrations of the n-6 FA linoleic acid (LeA), γ-linoleic acid, dihomo-γ-linoleic acid, and arachidonic acid were 1.428 ± 0.068, 0.070 ± 0.007, 0.066 ± 0.004, and 0.089 ± 0.004 g per 100 g of fat, respectively; adrenic acid was present in concentrations lower than 0.020 g per 100 g of fat, whereas docosapentaenoic acid was absent in all samples. The concentrations of ALA and LeA were significantly higher in spring and summer, compared with autumn and winter. The concentrations of all other ALA- and LeA-derived n-3 and n-6 FA were not significantly different between seasons. The contribution of milk fat to the daily intake of eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid was calculated for human consumption levels in different countries. Milk fat contributed between 10.7 and 14.1% to the daily intake of eicosapentaenoic acid and between 23.5 and 34.2% to the intake of docosapentaenoic acid; whereas docosahexaenoic acid contribution was marginal. Arachidonic acid from milk fat contributed between 10.5 and 18.8% to the human intake of n-6 FA.

Prediction of bulk milk fatty acid composition based on farming practices collected through on-farm surveys - Corrected Proof
M. Coppa, A. Ferlay, C. Chassaing, C. Agabriel, F. Glasser, Y. Chilliard, G. Borreani, R. Barcarolo, T. Baars, D. Kusche, O.M. Harstad, J. Verbič, J. Golecký, B. Martin

ABSTRACT: The aim of this study was to predict the fatty acid (FA) composition of bulk milk using data describing farming practices collected via on-farm surveys. The FA composition of 1,248 bulk cow milk samples and the related farming practices were collected from 20 experiments led in 10 different European countries at 44°N to 60°N latitude and sea level to 2,000 m altitude. Farming practice-based FA predictions [coefficient of determination (R2) >0.50] were good for C16:0, C17:0, saturated FA, polyunsaturated FA, and odd-chain FA, and very good (R2 ≥0.60) for trans-11 C18:1, trans-10 + trans-11 C18:1, cis-9,trans-11 conjugated linoleic acid, total trans FA, C18:3n-3, n-6:n-3 ratio, and branched-chain FA. Fatty acids were predicted by cow diet composition and by the altitude at which milk was produced, whereas animal-related factors (i.e., lactation stage, breed, milk yield, and proportion of primiparous cows in the herd) were not significant in any of the models. Proportion of fresh herbage in the cow diet was the main predictor, with the highest effect in almost all FA models. However, models built solely on conserved forage-derived samples gave good predictions for odd-chain FA, branched-chain FA, trans-10 C18:1 and C18:3n-3 (R2 ≥0.46, 0.54, 0.52, and 0.70, respectively). These prediction models could offer farmers a valuable tool to help improve the nutritional quality of the milk they produce.

Effect of estrus synchronization on daily somatic cell count variation in goats according to lactation number and udder health status - Corrected Proof
A. Mehdid, J.R. Díaz, A. Martí, G. Vidal, C. Peris

ABSTRACT: Two repeated experiments were carried out in 2 different years to study the effect of estrus on somatic cell count (SCC) in dairy goats. In the first year, 36 Murciano-Granadina goats were used [12 primiparous and 24 multiparous; 22 healthy and 14 with an intramammary infection (IMI)] and, after a 6-d pre-experimental period, were divided into 2 groups according to lactation number, udder health status, SCC, and milk production. One group was kept as a control, whereas the other received an estrus synchronization hormonal treatment lasting 11 d. At 24, 48, and 72 h after cessation of the hormone treatment, goats were placed in contact with a buck to confirm that they were in estrus. For 32 consecutive days (6 pre-experimental, 11 in hormone treatment, and 15 post-treatment) the SCC per gland and udder were monitored in all animals. In the second year, we repeated the same experimental design using a total of 38 Murciano-Granadina breed goats (12 primiparous and 26 multiparous; 26 healthy and 12 with IMI). Throughout this experiment, milk yield and composition were also recorded daily for each goat. Upon termination of the hormonal treatment, the SCC in udder milk increased significantly in the treatment group compared with the control group over 3 consecutive days. This increase was observed for year (1 and 2), parity (primiparous and multiparous), and udder health status (healthy and IMI). The log10 SCC (cells/mL) increased from 5.5 ± 0.09 before estrus to 6.04 ± 0.09 during treatment; therefore, the geometric mean of the SCC increased 3.5 times during treatment. The maximum values obtained in healthy glands of primiparous goats (geometric mean = 0.37 million cells/mL) were lower than in healthy glands (1.1 million cells/mL) or infected glands (1.7 million cells/mL) of multiparous goats. The increase in SCC observed during estrus (200% increase in geometric means) could not be explained by the changes in milk production, which only fell by 13%. During estrus, the percentage of protein and dry matter in the milk also increased significantly. We concluded that it is necessary to consider the presence of estrus to correctly interpret milk SCC, as an indirect method for detecting IMI or as a commercial milk quality parameter.

Sexually active groups in cattle—A novel estrus sign - Corrected Proof
G. Sveberg, A.O. Refsdal, H.W. Erhard, E. Kommisrud, M. Aldrin, I.F. Tvete, F. Buckley, A. Waldmann, E. Ropstad

ABSTRACT: The current study presents a novel objective measure for characterizing sexually active groups (SAG 3–5) and relates this measure to other behaviors of lactating Holstein-Friesian cows. Cows in SAG 3–5 were required to participate in a minimum of 1 estrus behavior per 5 min while staying within 3 m (2 cow lengths) of its partner(s) for a minimum of 5 min. Twenty Holstein-Friesian cows were video-monitored continuously through 1 complete estrous cycle (22 d). Standing behavior, SAG 3–5, secondary estrus signs (SEC), and other social and agonistic behaviors were recorded continuously. The period of mounting estrus (MTE) was divided into the 3 parts: prestand, standing estrus (STE), and poststand. The mean durations of MTE, prestand, STE, and poststand period were 12.9 ± 1.84, 4.0 ± 1.93, 7.1 ± 1.44, and 1.8 ± 0.57 h (n = 13). The fractions of time spent in SAG 3–5 during MTE, prestand, STE, and poststand period were 13, 8, 19, and 1% (n = 11). During MTE, cows participated, on average, in 5.8 ± 1.24 SAG 3–5 and initiated 9.5 ± 2.99 mounts, with mean durations of 0.25 ± 0.03 h and 4.00 ± 0.36 s, respectively. The novel measure SAG 3–5 was a sign of long duration not confined only to groups of STE cows. On one day when no cows were in estrus and during the periods 4 to 24 h before and after MTE, no SAG 3–5 behaviors were observed. Luteal-phase cows participated in SAG 3–5 only when the partner was a single cow in estrus. The time spent in SAG 3–5 increased between 1 and 3 h before MTE and the prestand period (3 vs. 8%) and reached a peak level during STE. From STE to poststand, time spent in SAG 3–5 decreased considerably (19 vs. 1%). The observed decrease in nonmutual agonistic behaviors 4 to 24 h before MTE is suggested as an early sign of pre-estrus. Changes in SAG 3–5, agonistic behaviors, and SEC are suggested as indicators of the specific stages of MTE. Increased SEC initiated and SAG 3–5 were indicators of late pre-estrus and early estrus (prestand). Peak levels of SAG 3–5, SEC, and social agonistic behaviors were indicators of STE. A sudden decrease in behaviors, preceded by frequent interactions, was indicative of late estrus (poststand). On the basis of the findings reported here, we propose that SAG 3–5, as well as proceptive and receptive patterns of SEC and agonistic behaviors, be included in estrus detection protocols. Updated knowledge of these behavioral interactions may assist when determining the stage of estrus and the optimal time to breed dairy cows.

Genetic associations of ketosis and displaced abomasum with milk production traits in early first lactation of Canadian Holsteins - Corrected Proof
A. Koeck, F. Miglior, J. Jamrozik, D.F. Kelton, F.S. Schenkel

ABSTRACT: The aim of this study was to investigate the genetic associations of ketosis and displaced abomasum with milk production traits in early first lactation of Canadian Holsteins. Health data recorded by producers were available from the national dairy cattle health system in Canada. Test-day records of milk, fat, and protein yields were obtained from the routine milk recoding scheme. Ketosis and displaced abomasum were defined as binary traits (0 = healthy; 1 = sick) based on whether or not the cow had at least 1 case of the respective disease in the period from calving to 100 d after calving. Mean frequencies of ketosis and displaced abomasum were 4.1 and 2.7%, respectively. The following milk production traits were considered: milk yield, fat percentage (Fat%), protein percentage (Prot%), fat-to-protein (F:P) ratio, and F:P ratio >1.5. The trait F:P ratio >1.5 was scored as 1 or 0, based on whether or not the cow had an F:P ratio >1.5. For milk production traits, the first (5–30 d in milk) and the second (31–60 d in milk) test days were considered. Data were analyzed using bivariate linear animal models. Average heritabilities of 0.02 and 0.04 were obtained for ketosis and displaced abomasum, respectively. For milk production traits, the lowest heritabilities were obtained for F:P >1.5 (0.04 to 0.08), whereas the highest estimates were found for Prot% (0.27 to 0.38). Ketosis and displaced abomasum were genetically uncorrelated with milk yield in early lactation. Moderate favorable correlations were found between metabolic diseases and milk composition traits. Ketosis was significantly correlated with Fat% (0.33), F:P ratio (0.30), and F:P ratio >1.5 (0.35) at the first test day, whereas all genetic correlations with milk composition traits at the second test day were not significant and close to zero. Significant favorable genetic correlations were also found between displaced abomasum and F:P ratio (0.26), F:P ratio >1.5 (0.25) and Prot% (−0.19) at the first test day. Also, Prot% at the second test day was significantly correlated (−0.16) with displaced abomasum. Overall, a higher Fat% and F:P ratio and a lower Prot% at the first test day were associated with an increased susceptibility to metabolic diseases. As genetic correlations between metabolic diseases and F:P ratio were far from unity, dairy producers should be encouraged to keep accurate and complete health data. This will be expected to yield to more accurate genetic evaluations for metabolic diseases.