Use of proximity loggers to Estimate Affiliative and Agonistic Relationships among Group-housed Holstein calves.

We used proximity loggers to map the social network of 15 group-housed suckling Holstein calves over a 70-day period divided into five trial periods. Tag ID, encountered tag ID, and encounter start time and length were collected from the proximity loggers. Use of 3D matrices and trendlines distinguished three encounter types: "rapid decline" (up to ~ 120 s), "moderate decline" (~120 to ~ 600 s), and "exponential decline" (longer than ~ 600 s). We hypothesized that a random encounter between two calves with an agonistic relationship would terminate quickly, whereas affiliative encounters would linger. Quadratic assignment with 5000 random permutations revealed a significant negative correlation between short encounters ("agonistic") and long encounters ("affiliative") matrices in periods 1 and 5 (r = -0.490 and -0.767, respectively), tendency toward negative correlation in period 3 (r = -0.141, p = 0.104), and no significant correlations in periods 2 and 4. These results were in accordance with the social network analyses revealing agonistic (short encounters) relationships in those periods. We suggest using this method in further studies on the effects of various factors on social network dynamics and its effect on health, intake, growth, and efficiency.

Re-Evaluation of Genotyping Methodologies in Cattle: The Proficiency of Imputation.

In dairy cattle, identifying polymorphisms that contribute to complex economical traits such as residual feed intake (RFI) is challenging and demands accurate genotyping. In this study, we compared imputed genotypes (n = 192 cows) to those obtained using the TaqMan and high-resolution melting (HRM) methods (n = 114 cows), for mutations in the FABP4 gene that had been suggested to have a large effect on RFI. Combining the whole genome sequence (n = 19 bulls) and the cows' BovineHD BeadChip allowed imputing genotypes for these mutations that were verified by Sanger sequencing, whereas, an error rate of 11.6% and 10.7% were encountered for HRM and TaqMan, respectively. We show that this error rate seriously affected the linkage-disequilibrium analysis that supported this gene candidacy over other BTA14 gene candidates. Thus, imputation produced superior genotypes and should also be regarded as a method of choice to validate the reliability of the genotypes obtained by other methodologies that are prone to genotyping errors due to technical conditions. These results support the view that RFI is a complex trait and that searching for the causative sequence variation underlying cattle RFI should await the development of statistical methods suitable to handle additive and epistatic interactions.

The Meat Quality Characteristics of Holstein Calves: The Story of Israeli 'Dairy Beef'.

Global animal production systems are often criticized for their lack of sustainability and insufficient resilience to ensure food security. The 'farm-to-fork' approach aims at orienting food systems towards the creation of a positive environmental impact, nutritious, healthy, safe and sufficient foods, and fairer economic returns for primary producers. Many countries rely on an imported supply of live animals to fulfill their needs for fresh meat. In Israel, ~60% of the sources of fresh beef come from the import of live animals. In order to encourage sustainable beef production in Israel, the proportion of local beef should be raised at the expense of imported animals. However, for this to be achieved, the superior performance of local beef should be justified. The current study was conducted to compare between the meat quality characteristics of local (Israeli Holstein; N = 205) vs. imported (Australian; N = 169) animals. Generally, while the imported calves presented a higher dressing percentage (p < 0.0001), the local animals were characterized by tenderer meat (p < 0.0001), longer sarcomeres (p < 0.0001), higher a* color attributes and pH (p < 0.001), superior cooking (p = 0.002) and thawing loss (p < 0.0001), higher intra-muscular fat (IMF) content, and a higher PUFA proportion (p < 0.01 and p < 0.0001, respectively) and PUFA:SFA ratio. The findings shown herein may provide sound arguments for stakeholders and policy makers to facilitate sustainable local beef production in Israel.

Dietary pomegranate peel improves milk quality of lactating ewes: Emphasis on milk fat globule membrane properties and antioxidative traits.

Concentrated pomegranate peel extract (CPE) was supplemented to ewes, and milk yield and fat content-fatty acid (FA) and phospholipid (PL) composition-were monitored. CPE-fed ewes had higher milk yield, and fat, protein and lactose contents than controls. Milk PL content-20% higher in the CPE-supplemented group-was regulated by treatment and not by total fat content; milk phosphatidylethanolamine and phosphatidylcholine increased by 22 and 26%, respectively, in CPE-supplemented vs. control ewes. Milk saturated FA concentration was higher, and total polyunsaturated and monounsaturated FA content lower in the CPE vs. control group, regardless of milk total fat content. CPE supplementation increased milk antioxidant capacity, suggesting antioxidant transfer from dietary source to milk, increasing stability and nutritive value. Our study provides first evidence for milk quality improvement in terms of antioxidants and PL enrichment without compromising total milk fat, suggesting strategies to improve dairy animals' milk composition without compromising total production.

FABP4 gene has a very large effect on feed efficiency in lactating Israeli Holstein cows.

Improving feed efficiency (FE) is a major goal for the livestock industry. Previously, we have identified 48 SNP markers distributed over 32 genes significantly associated with residual feed intake (RFI) in Israeli Holstein male calves, the most significant of which are located in the bovine FABP4 gene. In the present study, we tested associations of eight of the FABP4 markers with RFI and feed conversion ratio (FCR), along with milk composition and feeding behavioral traits, in 114 lactating Israeli Holstein cows. Large allele effects were found, along with large contributions of FABP4 markers to the phenotypic variation [mean contribution of all significant markers (P < 0.05), 15.4 and 12.0% for RFI and FCR, respectively] and genotypic variation [means of all significant markers (P < 0.05), 75.7 and 32.4% in RFI and FCR, respectively]. However, the association of all significant FABP4 markers with FE and milk content traits was found in opposite directions, such that improved FE was accompanied by decreased milk content. Hence, before inclusion in breeding programs, the gain in FE must be economically balanced with the loss in milk contents. On the other hand, these findings imply that in any current improvement program concentrated on milk traits alone, without taking into account the effect on FE, the progress in milk composition is probably accompanied by deterioration of FE. These results, if confirmed in other populations and breeds, set FABP4 as a prime candidate in any marker-assisted selection program targeting FE as a whole and RFI in particular.

The Use of Kosher Phenotyping for Mapping QTL Affecting Susceptibility to Bovine Respiratory Disease.

Bovine respiratory disease (BRD) is the leading cause of morbidity and mortality in feedlot cattle, caused by multiple pathogens that become more virulent in response to stress. As clinical signs often go undetected and various preventive strategies failed, identification of genes affecting BRD is essential for selection for resistance. Selective DNA pooling (SDP) was applied in a genome wide association study (GWAS) to map BRD QTLs in Israeli Holstein male calves. Kosher scoring of lung adhesions was used to allocate 122 and 62 animals to High (Glatt Kosher) and Low (Non-Kosher) resistant groups, respectively. Genotyping was performed using the Illumina BovineHD BeadChip according to the Infinium protocol. Moving average of -logP was used to map QTLs and Log drop was used to define their boundaries (QTLRs). The combined procedure was efficient for high resolution mapping. Nineteen QTLRs distributed over 13 autosomes were found, some overlapping previous studies. The QTLRs contain polymorphic functional and expression candidate genes to affect kosher status, with putative immunological and wound healing activities. Kosher phenotyping was shown to be a reliable means to map QTLs affecting BRD morbidity.

FABP4 is a leading candidate gene associated with residual feed intake in growing Holstein calves.

Ecological and economic concerns drive the need to improve feed utilization by domestic animals. Residual feed intake (RFI) is one of the most acceptable measures for feed efficiency (FE). However, phenotyping RFI-related traits is complex and expensive and requires special equipment. Advances in marker technology allow the development of various DNA-based selection tools. To assimilate these technologies for the benefit of RFI-based selection, reliable phenotypic measures are prerequisite. In the current study, we identified single nucleotide polymorphisms (SNPs) associated with RFI phenotypic consistency across different ages and diets (named RFI 1-3), using DNA samples of high or low RFI ranked Holstein calves. Using targeted sequencing of chromosomal regions associated with FE- and RFI-related traits, we identified 48 top SNPs significantly associated with at least one of three defined RFIs. Eleven of these SNPs were harbored by the fatty acid binding protein 4 (FABP4). While 10 significant SNPs found in FABP4 were common for RFI 1 and RFI 3, one SNP (FABP4_5; A

Adaptive traits of indigenous cattle breeds: The Mediterranean Baladi as a case study.

Generally taken, breeds of Bos taurus ancestry are considered more productive, in comparison with Bos indicus derived breeds that present enhanced hardiness and disease resistance, low nutritional requirements and higher capability of feed utilization. While breeds of B. taurus have been mostly selected for intensive production systems, indigenous cattle, developed mostly from indicine and African taurines, flourish in extensive habitats. Worldwide demographic and economic processes face animal production with new challenges - the increasing demand for animal food products. Intensification of animal husbandry is thus a desired goal in stricken parts of the world. An introduction of productive traits to indigenous breeds might serve to generate improved biological and economic efficiencies. For this to succeed, the genetic merit of traits like efficiency of feed utilization and product quality should be revealed, encouraging the conservation initiatives of indigenous cattle populations, many of which are already extinct and endangered. Moreover, to overcome potential genetic homogeneity, controlled breeding practices should be undertaken. The Baladi cattle are a native local breed found throughout the Mediterranean basin. Purebred Baladi animals are rapidly vanishing, as more European breeds are being introduced or used for backcrosses leading to improved production. The superiority of Baladi over large-framed cattle, in feedlot and on Mediterranean pasture, with respect to adaptability and efficiency, is highlighted in the current review.

Protein deprivation attenuates Hsp expression in fat tissue.

For ruminants, dietary protein is the first limiting component to the utilization of low-quality forage. Throughout gestation, low-protein intake may result in prenatal programming that causes various metabolic disturbances and physiological modulations to dams and their developing embryos. We studied the effect of long-term low-protein diet (LPD) on physiological, biochemical, and molecular parameters of the energy status in gestating beef cows. LPD resulted in significant reductions in feed intake and heart rate and promoted a negative retained energy status already after 3 weeks. Elevated levels of plasma creatinine and non-esterified fatty acids indicate endogenous degradation of fat and protein as a response to the demands in energy and nitrogen. Increasing levels of ß-hydroxybutyrate confirmed the negative energy status obtained by the physiological measurements. At the molecular level, subcutaneous fat, Hsp90, Hsp70, and proteasome subunits decreased significantly after 3 months on LPD, in parallel with an increase of adipocyte fatty acid-binding protein. These results may indicate a decrease in turn-over of proteins, at the cost of induced lipolysis, and suggest that the response to protein deprivation, when examined in an energy-storing tissue, includes downregulation of the constitutive heat shock proteins involved in the protein degradation pathway of energy production and upregulation of tissue-specific genes such as those involved in energy production from fat degradation.

Differential stress responses among newly received calves: variations in reductant capacity and Hsp gene expression.

Bovine respiratory disease complex (BRD), a major economic concern to the beef cattle industry all over the world, is triggered by physical, biological and psychological stresses. It is becoming noticeable that the key to reducing BRD appears to be centered at reducing the response to stress. The aims of the present study were to detect individual variations in the stress response of newly received young calves through their leukocyte heat shock protein (Hsp) response, selected neutrophil-related gene expression and oxidative stress, and relate them to pulmonary adhesions at slaughter, an indicative sign of clinical and subclinical episodes of BRD at an early age. Differential expression patterns of Hsp60 and Hsp70A1A were revealed in newly received calves 1 h, 5 h and 1 day after arrival, distinguishing between stress-responsive and non-stress-responsive individuals. Plasma cortisol was also indicative of stress-responsive and non-stress-responsive individuals, 1 h and 5 h after arrival. At the longer term, ß-glycan levels were highest 7 days after arrival and significantly correlated with an adhesion-free phenotype at slaughter. Oxidative stress responses, measured through the oxidation products of the exogenous linoleoyl tyrosine (LT) marker, revealed that hydroperoxidation and epoxidation of membranes may readily occur. Based on the LT oxidation products and levels of ß-glycan, we present a discriminant analysis model, according to which vulnerable individuals may be predicted at near 100% probability 7 days after arrival. Since clinical signs of BRD may often go undetected in feedlot calves, such a model, after its examination in large-scale experiments, may be a reliable tool for an early prediction of subclinical signs of BRD.

The potential of Pleurotus-treated olive mill solid waste as cattle feed.

The aims of the current study were to follow: (1) the capability of the edible mushroom Pleurotus ostreatus to degrade cell wall components and soluble phenols of the olive mill solid waste (OMSW), and improve it for ruminant nutrition (2) the fate of oil and the lipid-soluble compounds tocopherols, squalene and beta-sitosterol in the fermented OMSW. A significant decrease in oil and lipid-soluble compounds with a concomitant shift in the fatty acid profile and degradation of soluble phenols took place already after 14 d. The utilization of lipids by the fungus shifted the degradation of the structural carbohydrates to a later stage, and significantly reduced the metabolizable energy of the OMSW. We propose that edible fungi with reduced lipase activity would preserve the energy and health promoting ingredients of the oil, and force the fungus to degrade structural carbohydrates, thus improving its digestibility.

Caloric stress alters fat characteristics and Hsp70 expression in milk somatic cells of lactating beef cows.

Selection for higher production rate in cattle inhabiting challenging habitats may be considered disadvantageous because of possible deleterious effects on immunity and reproduction and, consequently, on calf crop percentage. In Israel, free-grazing high productive beef cows experience reduction in nutritional quality of forage during up to 8 months of the year. As milk production by dams dictates calf performance, dam's nutritional needs and rebreeding rates, the aim of the present study was to test how lactating beef cows deal with combined caloric and protein stress both at the productive and self protective levels. For this purpose, we studied the effect of long-term caloric stress on milk characteristics and gene expression of stress and milk components producing proteins. Lactating dams responded to caloric stress by decreased body weight, milk, and milk protein production. To compensate for total energy loses in milk, they produced milk of higher fat concentration and shifted the proportions of its fatty acids towards long and unsaturated ones. This was reflected by increased mRNA transcription of the fatty acid binding protein. Prolonged low-energy diet promoted cell-specific heat shock protein (Hsp) response; whereas significant increase of Hsp90 but unchanged levels of Hsp70 proteins were observed in white blood cells, the expression of Hsp70 in milk somatic cells was markedly attenuated, in parallel with a marked increase of alpha(s1)-casein expression. At the mammary gland level, these results may indicate a decrease in turnover of proteins and a shift to an exclusive expression of milk components producing factors. Similar responses to caloric stress were revealed also in ketotic dairy cows. Ketosis promoted a shift towards long and unsaturated fatty acids and an increased expression of alpha(s1)-casein in milk somatic cells. These findings may reflect an evolutionary-preserved mechanism in lactating cows for coping with caloric restriction. Overall, our results provide an index to test suitability of beef cattle breeds to inadequate caloric demands.

Nutritive and antioxidative potential of fresh and stored pomegranate industrial byproduct as a novel beef cattle feed.

Pomegranate peel is a nutritive-rich byproduct whose amounts are extensively growing due to the exponential increase in the production of pomegranate juice and "ready to eat" arils. Pomegranate peel is a rich source for antioxidants and thus may serve in the prevention of cattle diseases and in the improvement of beef products, making it an attractive component in beef cattle diets. The present study aims to evaluate the effect of commonly used storage practices on the nutritive and antioxidative properties of pomegranate peel. In general, storage conditions preserved most antioxidant capacity. Ensiling ambivalently affected the nutritive values of the peel and promoted increased levels of antioxidative components. In addition to polyphenols, nonphenolic components, such as alpha- and gamma-tocopherols, contributed to the total antioxidative capacity, and several minerals found in the peel added to its nutritional value. Dietary supplementation with fresh peels promoted significant increases in feed intake and alpha-tocopherol concentration in the plasma, with positive tendency toward increased weight gain of bull calves. All in all, the nutritive value and the antioxidant capacity of pomegranate peel turn it into a favorable health-promoting constituent of feedlot beef cattle diet.

The human sef-a isoform utilizes different mechanisms to regulate receptor tyrosine kinase signaling pathways and subsequent cell fate.

Negative feedback is among the key mechanisms for regulating receptor tyrosine kinase (RTK) signaling. Human Sef, a recently identified inhibitor of RTK signaling, encodes different isoforms, including a membrane spanning (hSef-a) and a cytosolic (hSef-b) isoform. Previously, we reported that hSef-b inhibited fibroblast proliferation and prevented the activation of mitogen-activated protein kinase (MAPK), without affecting protein kinase B/Akt or p38 MAPK. Conflicting results were reported concerning hSef-a inhibition of MAPK activation, and the effect of hSef-a on other RTK-induced signaling pathways is unknown. Here we show that, in fibroblasts, similar to hSef-b, ectopic expression of hSef-a inhibited fibroblast growth factor-induced cell proliferation. Unlike hSef-b, however, the growth arrest was mediated via a MAPK-independent mechanism, and was accompanied by elevated p38 MAPK phosphorylation and inhibition of protein kinase B/Akt. In addition, hSef-a, but not hSef-b, mediated apoptosis in fibroblast growth factor-stimulated cells. Chemical inhibitor of p38 MAPK abrogated the effect of hSef-a on apoptosis. In epithelial cells, ectopic expression of hSef-a inhibited the activation of MAPK, whereas down-regulation of endogenous hSef-a significantly increased MAPK activation and accelerated growth factor-dependent cell proliferation. These results indicate that hSef-a is a multifunctional negative modulator of RTK signaling and clearly demonstrate that hSef-a can inhibit the activation of MAPK, although in a cell type-specific manner. Moreover, the differences between the activities of hSef-a and hSef-b suggest that hSef isoforms can control signal specificity and subsequent cell fate by utilizing different mechanisms to modulate RTK signaling.

Reciprocal activation of HSF1 and HSF3 in brain and blood tissues: is redundancy developmentally related?

Transcriptional induction of heat-shock genes in response to temperature elevation and other stresses is mediated by heat-shock transcription factors (HSFs). Avian cells express two redundant heat-shock responsive factors, HSF1 and HSF3, which differ in their activation kinetics and threshold induction temperature. Unlike the ubiquitous activation of HSF1, the DNA-binding activity of HSF3 is restricted to undifferentiated avian cells and embryonic tissues. Herein, we report a reciprocal activation of HSF1 and HSF3 in vivo. Whereas HSF1 mediates transcriptional activity only in the brain upon severe heat shock, HSF3 is exclusively activated in blood cells upon light, moderate, and severe heat shock, promoting induction of heat-shock genes. Although not activated, HSF1 is expressed in blood cell nuclei in a granular appearance, suggesting regulation of genes other than heat-shock genes. Intraspecific comparison of heat-sensitive and heat-resistant fowl strains indicates that the unique activation pattern of HSF3 in blood tissue is a general phenomenon, not related to thermal history. Taken together, HSF1 and HSF3 mediate transcriptional activity of adult tissues and differentiated cells in a nonredundant manner. Instead, an exclusive, tissue-specific activation is observed, implying that redundancy may be developmentally related. The physiological and developmental implications are discussed.

Ectothermy and endothermy: evolutionary perspectives of thermoprotection by HSPs.

Living organisms respond to heat exposure by selectively expressing heat shock proteins (HSPs). Accumulation of HSPs confers thermotolerance in cell cultures and in ectotherms and is an important component of the heat shock response. This response, however, has not been directly examined in relation to different ;thermal states', namely ectothermy vs endothermy. By using avian development as a model system for transition from ectothermy to endothermy, we show that, in contrast to the ectothermic state, in the endothermic state the organism is more resistant to heat but relies less on HSPs as a first-line thermoprotective mechanism. Moreover, intraspecific, real-time, in vivo measurements in genetically diverse fowl strains relate improvement of thermoresistance in endotherms to improved body temperature (Tb) regulation, with a concomitant delay in the expression of HSPs. The time course of this delay and the Tb at which it occurs imply that the ontogenetic and evolutionary pathways leading to improved thermoresistance may have followed two, apparently non-related, parallel routes--cellular and peripheral (non-cellular). In search of other cellular components that differentially participate in the heat shock response, we revealed a significant expression of fatty acid synthase (FAS) in heat-exposed endotherms but not in ectotherms.

Alternative splicing generates an isoform of the human Sef gene with altered subcellular localization and specificity.

Receptor tyrosine kinases (RTKs) control a multitude of biological processes and are therefore subjected to multiple levels of regulation. Negative feedback is one of the mechanisms that provide an effective means to control RTK-mediated signaling. Sef has recently been identified as a specific antagonist of fibroblast growth factor (FGF) signaling in zebrafish and subsequently in mouse and human. Sef encodes a putative type I transmembrane protein that antagonizes the Ras/mitogen-activated protein kinase pathway in all three species. Mouse Sef was also shown to inhibit the phosphatidylinositol 3-kinase pathway. We show here that an alternative splicing mechanism generates an isoform of human Sef, hSef-b, which unlike the previously reported Sef (hSef-a) is a cytosolic protein. Contrary to hSef-a, which is ubiquitously expressed, hSef-b transcripts display a restricted pattern of expression in human tissues. hSef-b inhibits FGF-induced cell proliferation and prevents the activation of mitogen-activated protein kinase without affecting the upstream component MAPK kinase. Furthermore, hSef-b does not antagonize FGF induction of the phosphatidylinositol 3-kinase pathway. In addition to the effects on FGF signaling, hSef-b inhibited cellular response to platelet-derived growth factor but not other RTK ligands. Therefore, alternative splicing of the hSef gene expands the Sef feedback inhibition repertoire of RTK signaling.