Genome-wide association study for age at puberty and resumption of cyclicity in a crossbred dairy cattle population.

Fertility is of primary economic importance in dairy cattle and the most common reason for involuntary culling. However, standard fertility traits have very low heritability that renders genetic selection slow and difficult. In this study, we explored fertility from an endocrine standpoint. A total of 1,163 crossbred Holstein-Normande females in a 3-generation familial design were studied for progesterone level measured every 10 d to determine age at puberty (PUB) and commencement of postpartum luteal activity (CPLA). Genetic parameters were estimated using REML with WOMBAT software. The heritability estimates were 0.38 ± 0.10 and 0.16 ± 0.07 for PUB and CPLA, respectively. Moreover, the 2 traits were genetically correlated (0.45 ± 0.23), suggesting a partially common determinism. Because of the family structure, a linkage disequilibrium and linkage analysis approach was preferred over standard genome-wide association study to map genomic regions associated with these traits. Ten quantitative trait loci (QTL) were detected for PUB on chromosomes 1, 3, 11, 13, 14, 21, and 29, whereas 3 QTL were associated with CPLA on chromosomes 21 and 26. Only the QTL on chromosome 21 was common to both traits. Four functional candidate genes (NCOA2, GAS2, OVOL1, and FOSL1) were identified in the detected regions. These findings will contribute to a clearer understanding of fertility determinism and enhance the value of introducing endocrinological data in fertility studies.

Genetic parameters of milk production traits in response to a short once-daily milking period in crossbred Holstein × Normande dairy cows.

Despite its potential utility for predicting cows' milk yield responses to once-daily milking (ODM), the genetic basis of cow milk trait responses to ODM has been scarcely if ever described in the literature, especially for short ODM periods. This study set out to (1) estimate the genetic determinism of milk yield and composition during a 3-wk ODM period, (2) estimate the genetic determinism of milk yield responses (i.e., milk yield loss upon switching cows to ODM and milk yield recovery upon switching them back to twice-daily milking; TDM), and (3) seek predictors of milk yield responses to ODM, in particular using the first day of ODM. Our trial used 430 crossbred Holstein × Normande cows and comprised 3 successive periods: 1 wk of TDM (control), 3 wk of ODM, and 2 wk of TDM. Implementing ODM for 3 wk reduced milk yield by 27.5% on average, and after resuming TDM cows recovered on average 57% of the milk lost. Heritability estimates in the TDM control period and 3-wk ODM period were, respectively, 0.41 and 0.35 for milk yield, 0.66 and 0.61 for milk fat content, 0.60 and 0.80 for milk protein content, 0.66 and 0.36 for milk lactose content, and 0.20 and 0.15 for milk somatic cell score content. Milk yield and composition during 3-wk ODM and TDM periods were genetically close (within-trait genetic correlations between experimental periods all exceeding 0.80) but were genetically closer within the same milking frequency. Heritabilities of milk yield loss observed upon switching cows to ODM (0.39 and 0.34 for milk yield loss in kg/d and %, respectively) were moderate and similar to milk yield heritabilities. Milk yield recovery (kg/d) upon resuming TDM was a trait of high heritability (0.63). Because they are easy to measure, TDM milk yield and composition and milk yield responses on the first day of ODM were investigated as predictors of milk yield responses to a 3-wk ODM to easily detect animals that are well adapted to ODM. Twice-daily milking milk yield and composition were found to be partly genetically correlated with milk yield responses but not closely enough for practical application. With genetic correlations of 0.98 and 0.96 with 3-wk ODM milk yield losses (in kg/d and %, respectively), milk yield losses on the first day of ODM proved to be more accurate in predicting milk yield responses on longer term ODM than TDM milk yield.

Effects of breed, feeding system, and lactation stage on milk fat characteristics and spontaneous lipolysis in dairy cows.

Spontaneous lipolysis (SL) is an enzymatic reaction that leads to a release of fatty acids that can modify technological and sensory properties of milk and milk products. However, few studies have been done to assess the effect of feeding systems (FS) and breed on SL. Most of them were conducted in the 1980s and are not fully representative of cattle today. No previous study investigated the effect of cow breed at the whole-lactation scale. Thus, a trial was carried out to study the effects of 2 FS (high- and low-input FS) with 2 breeds [Holstein (HO) and Normande (NO)] during 1 entire lactation. Sixty-three cows were followed throughout 1 lactation. Cows were divided into 4 groups according to their breed and their FS. The high FS (HFS) consisted of a high-energy diet (in winter, corn silage with 30% concentrate; otherwise, pasture with 4 kg/d of concentrate) and the low FS (LFS) consisted of a low-energy diet (in winter, conserved grass with no concentrate; otherwise, pasture with no concentrate). The cows calved between January and March. Individual milk samples were collected every month from both morning and evening milkings for fat, protein, milk fat globule size, major fatty acids and proteins profiles, and SL determinations. Data were analyzed using the mixed procedure of SAS. The SL was higher in evening milks compared with morning milks. In early lactation, in evening milks, SL was higher in LFS than in HFS. No difference was shown according to the FS in mid and late lactation. Pasture was associated with low SL rate in mid lactation. The NO cows were less susceptible to SL during the entire lactation than HO cows. Finally, early and late lactation periods were identified as being more susceptible to SL, but this depended on breed and FS. During early lactation, HO cows and LFS were associated with higher levels of SL, particularly in evening milks, and, during late lactation, HO cows were associated with higher levels of SL. No intertreatment or intercow correlations (coefficient of determination <0.16) were found between SL, milk fat and protein contents, milk production, milk fat globule size, proportion of fatty acids and proteins, body condition, and weight during the entire lactation. Effects of breed, FS, and lactation stage were clearly identified and quantified. Causal mechanisms might involve energy balance and circadian secretion of milk fat globule components.

Effects of DGAT1 K232A polymorphism and milking frequency on milk composition and spontaneous lipolysis in dairy cows.

Milk spontaneous lipolysis (SL) of milk triglycerides is induced by the lipoprotein lipase, a milk native enzyme, and leads to an accumulation of partial glycerides and free fatty acids that are responsible for the deterioration of the taste of milk products. The gene coding for diacylglycerol acyltransferase 1 (DGAT1), an enzyme implicated in triglycerides synthesis, has an important polymorphic site at the K232A locus. This gene is well known to modulate milk composition. No data are available on the effects of DGAT1 on SL. Thus, a trial was carried out to evaluate the effects of DGAT1 K232A polymorphism on milk SL upon milking frequency variations [once- (ODM) and twice-daily milking (TDM)]. Twenty-one cows were divided into 3 groups according their DGAT1 K232A genotype: 8 cows had the KK genotype of DGAT1 (KK cows), 8 had the KA genotype (KA cows), and 5 had the AA genotype (AA cows). The trial consisted in 3 successive periods: 3 wk of TDM, 3 of ODM, and 4 of TDM. Samples were collected for fat and protein contents, SL, fatty acid, and protein profiles determinations. The KK cows presented higher fat and protein contents, lower milk production, and higher κ-casein percentage. No significant difference in fatty acid composition was noted between groups. The SL was twice as high for KK cows in TDM situations (1.13 vs. 0.59 and 0.63mEq/100g of fat, respectively, for KK, KA, and AA cows during the first period of TDM, and 0.46 vs. 0.25 and 0.21mEq/100g of fat, respectively, for KK, KA, and AA during the second period of TDM). The SL remained lower in TDM2 than in TDM1. During ODM, no difference in SL was found between groups and SL remained below 0.2mEq/100g of fat. These results demonstrate the existence of a correlation between DGAT1 genotypes and spontaneous lipolysis, in interaction with an environmental factor, milking frequency, although it has not been possible to clarify the causal mechanism at this stage.

Effect of dairy production system, breed and co-product handling methods on environmental impacts at farm level.

Six dairy farms with the same on-farm area and milk production were compared. One farm (G-No) used grass as the sole forage for a herd of Normande cows, a dual-purpose breed. Three farms, with Holstein cows, varied forage for the herd from grass only (G-Ho) to low (G/LM-Ho) or high (G/HM-Ho) proportion of maize silage in the total forage area. Finally, two farms based on G/LM-Ho and G/HM-Ho systems aimed to increase omega-3 fatty acids in the winter diets of cows (G/LM/O3-Ho, G/HM/O3-Ho). Allocation methods (biophysical, protein, economic allocation) and system expansion applied for co-product (milk and meat) handling were examined. The impact categories considered were climate change, climate change including the effects of land use and land use change (CC/LULUC), cumulative energy demand, eutrophication, acidification and land occupation. The impacts per kg of fat-and-protein-corrected milk (FPCM) of G-No were highest, followed by those of G-Ho, G/LM-Ho and G/HM-Ho, regardless co-product handling methods and impact categories (except for eutrophication). CC/LULUC per kg FPCM of G/LM/O3-Ho and G/HM/O3-Ho were both 1% and 3% lower than those of G/LM-Ho and G/HM-Ho, respectively, but other impacts were higher. With system expansion, impacts per kg FPCM were lower than when allocation methods were used. Enteric fermentation was the greatest contributor (45-50%) to CC/LULUC, while grass production was the most important contributor to other impacts. The highest CC/LULUC (for G-No) can be explained by (1) G-No having the lowest milk yield/cow (though it produced the most meat) and (2) the fact that grass required more N fertiliser, but had lower yields than silage maize, even though grassland sequestered C. Among Holstein systems, increasing cow productivity by increasing feed intake (including maize silage and supplementing with concentrate) decreased impacts of milk. Reducing replacement rate and age of first calving also decreased impacts of milk. Increasing cow productivity reduced the amount of on-farm area required to produce a given amount of milk. Thus, the "liberated" on-farm area of Holstein systems was used to produce cash crops, and total impacts of these systems were lower than those of G-No (except for eutrophication and land occupation).

Towards a better understanding of the respective effects of milk yield and body condition dynamics on reproduction in Holstein dairy cows.

The overall reproductive performance has decreased over the last decades, involving changes in cyclicity, oestrous behaviour and fertility. High milk yield (MY), low body condition score (BCS) and large body condition (BC) loss have been identified as risk factors. However, these effects are often confounded, as high MY and body lipid reserve mobilization are correlated. The aim of this study was to evaluate the respective effects of MY and BC on post-partum ovarian cyclicity, oestrus and fertility of Holstein cows. This study provides novel information, as MY and BC change were uncoupled in the overall dataset that included 98 lactations and milk progesterone profiles. Cows were assigned to two feeding-level groups: high feed, which achieved high MY and moderate BC loss throughout lactation (8410 kg, -1.17 unit from calving to nadir BCS), and low feed, which limited MY and triggered a large BC loss (5719 kg, -1.54 unit). MY and BC had different effects at different stages of the reproductive process. Cyclicity as well as non-fertilization and early embryo mortality were mainly driven by body lipid reserves, whereas oestrous behaviour and late embryo mortality were related to MY. The results point to possible uncoupling between cyclicity, oestrus and early and late embryo survival allowing compensation along the reproductive process and leading to similar final reproductive performance. In compact calving systems, which require high pregnancy rates within a short period, higher MY strategies appear unsuitable even where BCS is maintained, owing to depressed oestrous behaviour and probably increased late embryo mortality, which delays rebreeding. Similarly, strategies that compromise cyclicity and fertility by excessively low BCS are unsuitable.

Lactose in blood plasma and the ability of dairy cows to tolerate once-daily milking in terms of milk loss and milk recovery.

This experiment described the variability among cows with the aim of studying the ability of dairy cows to tolerate once-daily milking (ODM) in terms of milk losses and milk recoveries observed when cows are switched from twice- to once-daily milking and then back to twice-daily milking (TDM). It also aimed to investigate whether or not lactose in blood plasma, which indicates the mammary epithelium permeability, is correlated with milk losses and recoveries and, consequently, a potential candidate assessor of dairy cow tolerance to ODM. The study used 86 crossbred dairy cows (Holstein × Normande) split into 5 groups over 2 yr. The trial consisted of 3 successive periods: a 1-wk control period of TDM, then 3 wk of ODM, followed by 2 wk of TDM. Blood samples were collected 1 to 1.5h before the morning milking and 5 to 6h after milking on d -3, 0, 1, 3, 7, 21, 23, and 28, where d 0 is the last d of the control period. Milk losses measured as kilograms per day were higher in cows with highest control-period milk yields (r=-0.66). When expressed in relative terms (%), milk losses were weakly correlated with the control-period milk yield (r=-0.22). When switched back to TDM, cows recovered only 47% of the milk lost. Milk recovery (kg/d) was weakly correlated with control milk yield (r=0.34) and not correlated with ODM milk yield. Milk recovery was correlated with milk yield losses: the higher the milk yield losses (kg/d or %), the higher the milk recovery (kg/d; r=-0.59 and -0.52, respectively). Rate of milk recovery expressed as the quantity of milk recovered per kilogram of milk lost, was not correlated with milk loss expressed as a percentage. This means that to be considered well-adapted to ODM scheduling, a cow must be well adapted to milk loss, and be well adapted to milk recovery. Blood plasma lactose concentration (log(10)-transformed data) was not correlated with milk production levels in either control, ODM, or back-to-TDM periods. It was poorly correlated with milk loss but was positively correlated with milk recovery (kg/d) and rate of milk recovery. We conclude that blood plasma lactose needs to be coupled with other indicators for evaluating dairy cow tolerance to ODM.

Dairy cows' reproductive response to feeding level differs according to the reproductive stage and the breed.

Reproductive performance has decreased over the last decades in many dairy systems. This study aimed at comparing the effects of a high- and a low-feeding level on reproductive stages (cyclicity, oestrus and fertility) of Holstein and Normande cows in a grass-based winter compact calving system. High-fed cows received a total mixed ration composed of 55% maize silage, 15% dehydrated alfalfa pellets and 30% concentrate in winter and 4 kg/day concentrate supply at grazing. Low-fed cows only received 50% grass silage and 50% haylage in winter and no concentrate at grazing. Low-fed cows produced less milk over 44 weeks, but lost more body condition (BC) than high-fed ones (5207 v. 7457 kg, -1.28 v. -0.96 unit, P < 0.001). Normande cows produced less milk and lost less BC than Holstein ones (5596 v. 7068 kg, -0.89 v. -1.36 unit, P < 0.001). Post-partum ovarian activity was little affected by the feeding level. In both breeds, ovulation detection rate was higher in low-fed cows (74% v. 59%, P < 0.001) and detection relied more frequently on standing behaviour (67% v. 55%, P = 0.022). In both breeds, the recalving rate following the first and second inseminations was not significantly affected by the feeding level, although more non-fertilisations or early embryo mortalities occurred in low-fed cows (43% v. 26%, P = 0.004). For the Holstein breed, this was clearly explained by more late embryo mortalities in high-fed cows (30% v. 9%, P = 0.004). Finally, pregnancy rate by the end of the 13-week breeding period was similar between feeding groups. Conversely, the Normande dual-purpose cows had a higher pregnancy rate by the end of the breeding period than the Holstein dairy cows (72% v. 54%, P = 0.007), owing to a better ovarian activity (79% v. 54% normal, P < 0.001) and a higher recalving rate following insemination (53% v. 37%, P = 0.007). In conclusion, this study demonstrates that feeding levels with converse effects on milk yield and BC score also have converse effects on reproductive stages and lead to quite similar final reproductive performance in compact calving systems. Normande cows benefit from an overall better reproductive performance, but do not suit high feeding levels for very compact calving systems owing to depressed oestrous behaviour.

Identification of a doublet missense substitution in the bovine LRP4 gene as a candidate causal mutation for syndactyly in Holstein cattle.

Syndactyly in Holstein cattle is an autosomal recessive abnormality characterized by the fusion of the functional digits. This disorder has been previously mapped to the telomeric part of bovine chromosome 15. Here, we describe the fine-mapping of syndactyly in Holstein cattle to a 3.5-Mb critical interval using a comparative mapping approach and an extended pedigree generated by embryo transfer. We report genetic evidence for the exclusion of two genes previously suggested as candidates (EXT2 and ALX4) and describe the identification of a doublet mutation in complete linkage disequilibrium with syndactyly in one gene of the critical interval: LRP4. Finally, based on recent discoveries concerning the mouse mutants dan and mdig and a mouse knockout for Lrp4, we present solid evidence that the subsequent substitution in LRP4 exon 33 is a strong candidate causal mutation for syndactyly in Holstein cattle.