The APOB loss-of-function mutation of Holstein dairy cattle does not cause a deficiency of cholesterol but decreases the capacity for cholesterol transport in circulation.

The loss-of-function mutation of the apolipoprotein (APO) B gene (APOB) in Holstein cattle accounts for increased losses in calves that are homozygous for this mutation. Heterozygous carriers of the APOB mutation are clinically healthy but show decreased concentrations of plasma cholesterol and lipoproteins. So far, the metabolic effects of the mutation have only been investigated in heterozygous calves, bulls, and nonlactating females. In high-yielding dairy cows, a marked decrease in cholesterol concentration in plasma during early lactation is part of the usual metabolic changes. Given the essential role of cholesterol in fatty acid and lipid metabolism, a specific effect of the APOB mutation on metabolism and performance in dairy cows is expected. Therefore, the aim of the present study was to investigate the effects of different APOB genotypes on metabolic parameters, hepatic metabolism, and lactation and reproductive performance. Twenty pairs of full siblings with similar age, performance, and calving were investigated. Both animals of each pair were kept on the same farm and consisted of a heterozygous carrier (CDC) and a noncarrier (CDF) of the APOB mutation associated with cholesterol deficiency. Blood samples were taken in early (25.5 ± 4.7 d in milk) and mid lactation (158.2 ± 11.1 d in milk; mean ± SD), and analyzed for nonesterified fatty acids, ß-hydroxybutyrate, glucose, insulin-like growth factor-1, aspartate aminotransferase and gamma-glutamyltransferase activity, total cholesterol, free cholesterol, triacylglycerols, high density lipoprotein-cholesterol, and phospholipids. The evaluation of milk production, milk gross composition, and lactation persistency was based on official Dairy Herd Improvement Association recordings. Cholesterol and lipoprotein concentrations were lower in CDC cows than in CDF cows in early and mid lactation. Metabolic parameters, triacylglycerol concentration in plasma, and lactation and reproductive performance did not differ between CDC cows and CDF cows. The low cholesterol concentrations associated with the APOB mutation in heterozygous carriers are not because of a primary deficiency of cholesterol at a cellular level, as the term "cholesterol deficiency" suggests, but rather a consequence of reduced capacity for its transport in circulation. Overall, the data of the present study suggest that, despite the presence of the APOB mutation, cholesterol is not limiting for animals' metabolic adaptation and performance in heterozygous Holstein cows.

Genome-wide mapping of the dominance effects based on breed ancestry for semen traits in admixed Swiss Fleckvieh bulls.

Heterosis is the beneficial deviation of crossbred progeny from the average of parental lines for a particular trait. Heterosis is due to nonadditive genetic effects with dominance and epistatic components. Recent advances in genotyping technology have encouraged researchers to estimate and scan heterosis components for a range of traits in crossbred populations, applying various definitions of such components. In this study, we defined the intralocus (dominance) component of heterosis using local genetic ancestry and performed genome-wide association analysis for admixed Swiss Fleckvieh bulls and their parental populations, Red Holstein Friesian and Swiss Simmental, for semen traits. A linear mixed model for 41,824 SNP, including SNP additive genetic, breed additive, and breed dominance effects on 1,178 bulls (148 Red Holstein Friesian, 213 Swiss Simmental, and 817 Swiss Fleckvieh) with a total of 43,782 measurements was performed. In total, 19 significant regions for breed dominance were identified for volume (2 regions on Bos taurus autosome 10 and 22) and percentage of live spermatozoa (17 regions on Bos taurus autosome 3, 4, 5, 7, 13, 14, and 17), and genes associated with spermatogenesis, sperm motility, and male fertility traits were located there. No significant region for breed dominance was detected for total number of spermatozoa. The signals for breed dominance were relatively wide, most likely due to limited numbers of recombination events in a small number of generations (10-15 generations) of crossbreeding in the recent Swiss Fleckvieh composite.

Effects of breed proportion and components of heterosis for semen traits in a composite cattle breed.

The aim of this study was to estimate the non-additive genetic effects of the dominance component of heterosis as well as epistatic loss on semen traits in admixed Swiss Fleckvieh, a composite of Simmental (SI) and Red Holstein Friesian (RHF) cattle. Heterosis is the additional gain in productivity or fitness of cross-bred progeny over the mid-purebred parental populations. Intralocus gene interaction usually has a positive effect, while epistatic loss generally reduces productivity or fitness due to lack of evolutionarily established interactions of genes from different breeds. Genotypic data on 38,205 SNP of 818 admixed, as well as 148 RHF and 213 SI bulls as the parental breeds were used to predict breed origin of alleles. The genomewide locus-specific breed ancestries of individuals were used to calculate effects of breed difference as well as the dominance component of heterosis, while proxies for two definitions of epistatic loss were derived from 100,000 random pairs of loci. The average Holstein Friesian ancestry in admixed bulls was estimated 0.82. Results of fitting different linear mixed models showed including the dominance component of heterosis considerably improved the model adequacy for three of the four traits. Inclusion of epistatic loss increased the accuracy of the models only for our new definition of the epistatic effect for two traits, while the other definition was so highly correlated with the dominance component that statistical separation was impossible.

Liver transcriptome analysis reveals important factors involved in the metabolic adaptation of the transition cow.

During early lactation, dairy cows experience a severe metabolic load often resulting in the development of various diseases. The inevitable deficiency in nutrients and energy at the onset of lactation requires an optimal adaptation of the hepatic metabolism to overcome metabolic stress. We conducted a whole-liver transcriptome analysis for the transition cow to identify novel factors crucial for metabolic adaptation. Liver samples were obtained from 6 Red Holstein dairy cows (parity 2 to 7, mean ± standard deviation: 3.7 ± 2.3) at 3 time points: T1 = 22 ± 4 d antepartum, T2 = 10 ± 2 d postpartum, and T3 = 17 ± 2 d postpartum. Using RNA sequencing (RNA-seq), we studied the transcriptomic profile of the transition cow before and after parturition. We performed a differential gene expression analysis (DGEA) and gene-set enrichment analysis (GSEA) for biological processes (gene ontology, GO) and pathways (Kyoto Encyclopedia of Genes and Genomes, KEGG). Among the 10,186 expressed genes, we discovered 1,063 differentially expressed genes (false discovery rate = 5%). The GSEA revealed 16 biological processes and 7 pathways significantly (false discovery rate = 5%) associated with the hepatic changes of the transition cow. Our results confirm that major hepatic changes are related to energy mobilization after parturition; in particular, they are related to fatty acid oxidation/metabolism, cholesterol metabolism, and gluconeogenesis. Using the STRING database (https://string-db.org/), we investigated interactions between significant genes and identified 9 key genes (CYP7A1, APOA1, CREM, LOC522146, CYP2C87, HMGCR, FDFT1, SGLE, and CYP26A1) through which the different processes involved in the metabolic adaptation interact. Comparing our main results with the literature, we could identify further genes that have not yet been associated with the transition period (e.g., CPT1B, ADIPOR2, LEPR, CREB3L3, and CCND1) and that are mainly involved in processes controlled by AMP-activated protein kinase, an important regulator of energy homeostasis.

A reaction norm sire model to study the effect of metabolic challenge in early lactation on the functional longevity of dairy cows.

Due to the discrepancy of the high energy demand for rapidly increasing milk production and limited feed intake in the transition period around parturition, dairy cows require considerable metabolic adaptations. We hypothesize that some cows are genetically less suited to cope with these metabolic needs than others, leading to adverse follow-up effects on longevity. To test this, we designed a reaction norm model in which functional lifetime was linked to the metabolic challenge in the beginning of the first lactation. As challenge variables, we used either the sum of milk yield or the accumulated fat-to-protein ratio of the first 3 test-days (<120 d in milk), pre-adjusted for herd-test-day variance. We defined a random regression sire model, in which a random slope was estimated for each sire to assess whether a bull had robust (neutral or positive slopes) or non-robust (negative slopes) daughters. We fitted the model to data of ∼580,000 daughters of ∼5,000 Brown Swiss bulls with suitable observations available (≥10 daughters per bull). To validate our proposed model and assess the reliability of the estimated (co)variance components, we conducted an extensive bootstrap approach. For both challenge variables, we found the sire variance for the slope of the random regression to be significantly different from zero, suggesting a genetic component for metabolic adaptability. The results of the study show that the ability to cope with metabolic stress in the transition period has a genetic component, which can be used to breed metabolically robust dairy cows.

Metabolic adaptation recorded during one lactation does not allow predicting longevity in dairy cows.

INTRODUCTION: Metabolic and health disorders account distinctly for culling in dairy cows. This study investigated if metabolic status obtained once in life during a negative energy balance in early lactation allows to predict age and lifetime performance animals achieved at culling. Metabolically stressed cows entering at least their 3rd lactation (n = 200, parity: 5.0 ± 2.1, mean ± SD) were selected from a field study conducted in Switzerland. Age of cows at culling ranged from 4.7 to 20.2 years with parities from 3 to 17. From cows with known reasons of culling, 28.4% were culled because of fertility, 16.4% due to udder health, 15.5% due to high age and 10.4% because of claw health/lameness. A retrospective classification of the one-time recorded metabolic adaptation in week 4 post partum did not differ between animals of different parities at culling. Furthermore, there was no relationship neither between the metabolic adaptation recorded in a preceding lactation and the number of lactations achieved, nor to the lifetime milk production. Contrary to the wide spread assumptions, an inadequate adaptation due to a high metabolic load in early lactation may not result in an earlier culling of dairy cows, although they are more prone to metabolic disorders.

INTRODUCTION: Les problèmes métaboliques ainsi que d'autres pathologies jouent un rôle important dans les causes de réforme des vaches laitières. Dans la présente étude, on a cherché à savoir si le statut métabolique, qui a été noté une fois durant un bilan énergétique négatif au début de la lactation, permettait de tirer des conclusions quant à l'âge et à la performance de vie au moment de la sortie de l'exploitation. On a choisi, dans le cadre d'une étude de terrain effectuée en Suisse, des vache surchargées métaboliquement à partir de la troisième lactation (n = 200, numéro de lactation: 5.0 ± 2.1, moyenne ± déviation standard). L'âge des vaches lors de la réforme variait entre 4.7 et 20.2 ans et ces vaches comptaient entre 3 et 17 lactations. Chez les vaches dont on connaissait la cause de réforme, celle-ci était pour 28.4% des troubles de fertilité, pour 16.4% des problèmes de mamelle, pour 15.5% un âge avancé et pour 10.4% des affections des onglons ou des boiteries. Une classification rétrospective basée sur la charge métabolique dans la quatrième semaine de lactation n'a pas montré de différence quant au nombre de lactations atteint ni quant à la performance de vie. Contrairement à l'idée répandue, on a pu montrer qu'une adaptation insuffisante due à une charge métabolique élevée en début de lactation ne conduisait pas forcément à une réforme précoce, bien que de telles vaches soient soumises à un risque plus élevé d'affections métaboliques.

Rapid Communication: Cholesterol deficiency-associated APOB mutation impacts lipid metabolism in Holstein calves and breeding bulls.

During the last months, the number of reports on Holstein calves suffering from incurable idiopathic diarrhea dramatically increased. Affected calves showed severe hypocholesterolemia and mostly died within days up to a few months after birth. This new autosomal monogenic recessive inherited fat metabolism disorder, termed cholesterol deficiency (CD), is caused by a loss of function mutation of the bovine gene. The objective of the present study was to investigate specific components of lipid metabolism in 6 homozygous for the mutation (CDS) and 6 normal Holstein calves with different genotypes. Independent of sex, CDS had significantly lower plasma concentrations of total cholesterol (TC), free cholesterol (FC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), very-low-density lipoprotein cholesterol (VLDL-C), triacylglycerides (TAG), and phospholipids (PL) compared with homozygous wild-type calves ( < 0.05). Furthermore, we studied the effect of the genotype on cholesterol metabolism in adult Holstein breeding bulls of Swissgenetics. Among a total of 254 adult males, the homozygous mutant genotype was absent, 36 bulls were heterozygous carriers (CDC), and 218 bulls were homozygous wild-type (CDF). In CDC bulls, plasma concentrations of TC, FC, HDL-C, LDL-C, VLDL-C, TAG, and PL were lower compared with CDF bulls ( < 0.05). The ratios of FC:cholesteryl esters (CE) and FC:TC were higher in CDC bulls compared with CDF bulls, whereas the ratio of CE:TC was lower in CDC bulls compared with CDF bulls ( < 0.01). In conclusion, the CD-associated mutation was shown to affect lipid metabolism in affected Holstein calves and adult breeding bulls. Besides cholesterol, the concentrations of PL, TAG, and lipoproteins also were distinctly reduced in homozygous and heterozygous carriers of the mutation. Beyond malabsorption of dietary lipids, deleterious effects of apolipoprotein B deficiency on hepatic lipid metabolism, steroid biosynthesis, and cell membrane function can be expected, which may result in unspecific symptoms of reduced fertility, growth, and health.

A transposable element insertion in APOB causes cholesterol deficiency in Holstein cattle.

Cholesterol deficiency, a new autosomal recessive inherited genetic defect in Holstein cattle, has been recently reported to have an influence on the rearing success of calves. The affected animals show unresponsive diarrhea accompanied by hypocholesterolemia and usually die within the first weeks or months of life. Here, we show that whole genome sequencing combined with the knowledge about the pedigree and inbreeding status of a livestock population facilitates the identification of the causative mutation. We resequenced the entire genomes of an affected calf and a healthy partially inbred male carrying one copy of the critical 2.24-Mb chromosome 11 segment in its ancestral state and one copy of the same segment with the cholesterol deficiency mutation. We detected a single structural variant, homozygous in the affected case and heterozygous in the non-affected carrier male. The genetic makeup of this key animal provides extremely strong support for the causality of this mutation. The mutation represents a 1.3kb insertion of a transposable LTR element (ERV2-1) in the coding sequence of the APOB gene, which leads to truncated transcripts and aberrant splicing. This finding was further supported by RNA sequencing of the liver transcriptome of an affected calf. The encoded apolipoprotein B is an essential apolipoprotein on chylomicrons and low-density lipoproteins, and therefore, the mutation represents a loss of function mutation similar to autosomal recessive inherited familial hypobetalipoproteinemia-1 (FHBL1) in humans. Our findings provide a direct gene test to improve selection against this deleterious mutation in Holstein cattle.