Associations between maternal milk protein genotypes with preweaning calf growth traits in beef cattle

The objectives of this study were to investigate milk casein polymorphisms in dams and to determine the impacts of maternal casein genotypes on growth traits of their sucking calves. Milk samples from 433 dams of the breeds German Angus (GA) and German Simmental (GS) were typed at the milk protein loci α _s1-casein (α_s1-CN), β-casein (β-CN), α _s2-casein (α_s2-CN), and κ-casein (κ-CN) via isoelectric focusing. Associations between casein genotypes in maternal milk with growth traits of their 1,872 calves were analyzed until the age of weaning using linear mixed models, considering either genotypes of individual casein loci (model 1) or composite α _s1-β-α _s2-κ-CN genotypes within the casein cluster (model 2). Besides environmental effects such as sex, age of the dam, and calving year-season, genetic effects (breed group and maternal and paternal effects) were considered in statistical models. The composite casein genotype BBǀA2A2ǀAAǀAB (order of genes on bovine chromosome 6: α _s1-ǀβ-ǀα _s2-ǀκ-CN) was associated with greater average daily weight gains (ADG) and heavier age-adjusted weaning weights (WW) of calves (P < 0.05). The effects of composite genotypes on birth weight of calves were similar (P > 0.05; model 2). With regard to individual casein loci, greater ADG and WW were observed for calves from dams with the genotypes κ-CN BB and α _s1-CN BB, respectively (P < 0.05; model 1). Age-adjusted WW was largest for calves from dams carrying the κ-CN genotype BB (215 kg) compared with calves representing the maternal AB and AA genotypes (both 204 kg). Results from the present study suggested selectable casein genotypes due to their nutritional value of milk (value in terms of offspring performances), offering new perspectives for breeding strategies in beef cattle to improve preweaning calf performance.     

Comparison of milk protein composition in a Swedish and a Danish dairy herd using reversed phase HPLC

Abstract

The milk composition in a Swedish herd, consisting of Swedish Red and White cows (SRB) and Swedish Holstein cows (SLB), and in a Danish herd, consisting of Danish Holstein-Friesian cows (SDM), was evaluated. Concentrations of the major milk proteins (κ-casein, α_S1-casein, β-casein, β-lactoglobulin A, β-lactoglobulin B and α-lactalbumin) of 134 individual milk samples were determined by reversed phase (RP) HPLC. Other parameters determined included milk fat, urea, lactose, calcium, lactoferrin, somatic cell count and protein degradation (determined as level of free amino-terminals). Analysis of variance was used to compare concentrations of analysed milk variables between SRB and SLB or between SLB and SDM. Concentration of total protein, total casein, β-casein and κ-casein were significantly higher in SRB milk compared with SLB milk. Concentration of α-lactalbumin and calcium were significantly higher in SDM milk than in SLB milk. The concentration of urea was higher in SLB than in SDM milk and is suggested to reflect differences in feeding regimes between the investigated Swedish and Danish herd.     

A2 β-酪蛋白消化特性与人体健康

β-酪蛋白是牛乳酪蛋白的重要组成成分,具有遗传多态性,特别是A1和A2型与人体健康密切相关。本文综述了A2型β-酪蛋白结构和功能的特性关系、消化特性的人群差异性及不同种类乳β-酪蛋白的消化特性,从消化特性的角度论述了影响人体健康的因素,旨在为不同人群开发具有良好消化特性的乳制品提供参考。     

IgE reactivity to milk components in dogs with cutaneous adverse food reactions

We investigated the IgE reactivity to crude and purified milk antigens in the sera of 112 dogs with cutaneous adverse food reactions (CAFRs). Of the 112 dogs, 33 (29%) had specific IgE for crude milk antigens. In the dogs with milk-specific IgE, IgE reactivity to casein, bovine serum albumin (BSA), α-lactalbumin, β-lactoglobulin, and bovine IgG were 81%, 85%, 39%, 27%, and 35%, respectively. Casein and BSA may be important allergens in dogs with CAFRs. Some canine vaccines contain casein hydrolysate as a stabilizer and the pooled serum with anti-casein IgE showed IgE reactivity to the vaccines containing it. Information about IgE reactivity to casein in dogs with CAFRs could be useful for predicting adverse reactions to the vaccines including casein hydrolysate.     

Necrotizing Infectious Enteritis in Piglets,Caused by Clostridium Perfringens Type C: I. Biochemical and Toxigenic Properties of the Clostridium

Twenty Danish and 3 foreign strains of the porcine subtype of Cl. perfringens type G have been studied with regard to biochemical activity and toxin formation.The strains examined showed no deviation from the biochemical features characteristic of Gl. perfringens. Two of the strains showed delayed fermentation of saccharose, 1 of them also of salicin. Only 3 of the strains gave a positive nitrite reaction. The rest of the strains (20) presumably reduced nitrate to NH₃ within 3 days.All of the strains examined produced the major antigens α and β, none of them ε and ι. Of the minor antigens, θ and v were produced by all the strains, x by 2 of the foreign strains and 5 of the Danish strains. None of the strains gave reaction for the δ, λ and μ antigens.Some of the Danish strains were hemolytic in the presence of α, δ, and θ antitoxins, others not, while a few varied. The foreign strains were non-αδθ hemolytic.     

乳制品中A1 β-酪蛋白、A2 β-酪蛋白检测方法研究进展

随着消费者对A2 β-酪蛋白的关注度提升,A2 β-酪蛋白相关产品不断更新换代。A2 β-酪蛋白常用检测方法有反相高效液相色谱法（RP-HPLC）、液相色谱–高分辨串联质谱法、毛细管区带电泳法、试剂盒检测法等。本文对β-酪蛋白遗传变体A1 β-酪蛋白和原始形态A2 β-酪蛋白的功能特点、检测方法、定量分析进行综述,说明检测方法的原理和试剂药品的作用机理,加强对A1 β-酪蛋白、A2 β-酪蛋白检测方法的改进与优化,为A2 β-酪蛋白应用于功能性食品、婴幼儿配方食品提供理论依据。     

β-Lactoglobulin Phenotypes in Finnish Ayrshire and Friesian Cattle with Special Reference to Mastitis Indicators

An unselected material consisting of composite milk samples from 1029 Ayrshire and 113 Friesian cows were analysed for the β-lactoglobulin types. The frequencies of β-Lg types were AA 8.3 %, AB 45.5% and BB 46.3 % for Ayrshire cows and 22.1 %, 45.1 % and 32.8 % for Friesian cows, respectively. The relationship between β-Lg type with milk BSA, somatic cell count, protein percentage, fat percentage and milk production were analysed. AA cows were significantly higher in daily milk yield but lower in protein percentage and fat percentage than BB cows; AB types were intermediate. The annual production was highest in AB-animals. There was a tendency for AA cows to have high somatic cell counts but low milk BSA concentrations.β-lactoglobulins; milk BSA; somatic cell count; bovine mastitis; bovine milk; milk proteins.     

九龙藏黄牛乳蛋白的组成及遗传多态性研究

本研究旨在了解生活在高海拔地区的藏黄牛乳的生化组成特点。试验测定了43头九龙藏黄牛乳蛋白含量、乳蛋白组成和酪蛋白多态性。藏黄牛乳蛋白含量低,仅约为29 g/L;电泳分析结果显示,脱脂乳蛋白组成与普通牛乳接近,主要包括酪蛋白、α-乳清蛋白和β-乳球蛋白等组分,β-Lg与α-La含量的比值低于牦牛;酪蛋白的相对含量约为70%,主要包含α_s-CN和β-CN,二者分别检测到3和2种基因型。研究结果表明,九龙藏黄牛乳的组成与生活在类似生态环境中的九龙牦牛存在差异。     

羊乳蛋白质组成及消化特性

应用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和体外模拟消化研究羊乳蛋白质组成和消化特性。结果表明：羊乳蛋白质主要由酪蛋白和乳清蛋白组成,酪蛋白主要由αs1-酪蛋白、αs2-酪蛋白、β-酪蛋白和κ-酪蛋白组成,在酪蛋白中的相对含量分别为23.10%、30.39%、38.09%和8.42%;乳清蛋白主要由α-乳白蛋白、β-乳球蛋白、乳铁蛋白、血清白蛋白和免疫球蛋白组成,在乳清蛋白中的相对含量分别为24.59%、57.50%、4.35%、8.69%和4.88%;羊乳酪蛋白主要在肠液中消化,在胃液中消化120 min时     

Characterization of β-adrenergic receptors in bovine intramuscular and subcutaneous adipose tissue: comparison of lubabegron fumarate with β-adrenergic receptor agonists and antagonists

Chinese hamster ovary cell constructs expressing either the β ₁-, β ₂- or β ₃-adrenergic receptor (AR) were used to determine whether a novel β-AR modulator, lubabegron fumarate (LUB; Experior, Elanco Animal Health) might exert greater potency for a specific β-AR subtype. EC₅₀ values calculated based on cAMP accumulation in dose response curves indicate that LUB is highly selective for the β ₃-AR subtype, with an EC₅₀ of 6 × 10^–9 M, with no detectible agonistic activity at the β ₂-AR. We hypothesized that the accumulation of lipolytic markers would reflect the agonist activity at each of the β-receptor subtypes of the specific ligand; additionally, there would be differences in receptor subtype expression in subcutaneous (s.c.) and intrmuscular (i.m.) adipose tissues. Total RNA was extracted from adipose tissue samples and relative mRNA levels for β ₁-, β₂-, and β ₃-AR were measured using real-time quantitative polymerase chain reaction. Fresh s.c. and i.m. adipose tissue explants were incubated with isoproterenol hydrochloride (ISO; β-AR pan-agonist), dobutamine hydrochloride (DOB; specific β ₁-AA), salbutamol sulfate (SAL; specific β ₂-AA), ractopamine hydrochloride (RAC), zilpaterol hydrochloride (ZIL), BRL-37344 (specific β ₃-agonist), or LUB for 30 min following preincubation with theophylline (inhibitor of phosphodiesterase). Relative mRNA amounts for β ₁-, β ₂-, and β ₃-AR were greater (P < 0.05) in s.c. than in i.m. adipose tissue. The most abundant β-AR mRNA in both adipose tissues was the β ₂-AR (P < 0.05), with the β ₁- and β ₃-AR subtypes being minimally expressed in i.m. adipose tissue. ISO, RH, and ZH stimulated the release of glycerol and nonesterified fatty acid (NEFA) from s.c. adipose tissue, but these β-AR ligands did not alter concentrations of these lipolytic markers in i.m. adipose tissue. LUB did not affect glycerol or NEFA concentrations in s.c. or i.m. adipose tissue, but attenuated (P < 0.05) the accumulation of cAMP mediated by the β ₁- and β ₂-AR ligands DOB and SAL in s.c. adipose tissue. Collectively, these data indicate that bovine i.m. adipose tissue is less responsive than s.c. adipose tissue to β-adrenergic ligands, especially those that are agonists at the β ₁- and β₃-receptor subtypes. The minimal mRNA expression of the β ₁- and β ₃ subtypes in i.m. adipose tissue likely limits the response potential to agonists for these β-AR subtypes.     

Effect of storage time and temperature on the total protein concentration and electrophoretic fractions in equine serum

Serum protein electrophoresis (SPE) is a technique that could be considered one of the most useful diagnostic aids available to the clinician. The effect of storage time and temperature on the total proteins and electrophoretic fractions (albumin, α₁-, α₂-, β₁-, β₂-, and γ-globulins) was assessed in 24 healthy horses. All samples, collected by jugular vein puncture, were centrifuged and divided into 4 aliquots. The 1st aliquot was analyzed within 3 h from collection (time 0), the 2nd was refrigerated at +4°C for 24 h, the 3rd was refrigerated at +4°C for 48 h, and the last was frozen at −20°C for 48 h. One-way repeated-measures analysis of variance (ANOVA) showed a significant effect (P < 0.05) of the different storage conditions on the concentrations of all the parameters studied and significant variations in the percentages of albumin, α₁-globulins, α₂-globulins, and γ-globulins. Compared with time 0 the total protein concentration increased significantly after 48 h at −20°C, the albumin percentage decreased after 48 h at −20°C, the α₁-globulin percentage increased after 24 h at +4°C, the α₂-globulin percentage increased after 48 h at +4°C and at −20°C, and the γ-globulin percentage increased after 48 h at −20°C. The results should help veterinary practitioners handle and store equine serum samples appropriately. Further investigations at different storage times and temperatures could be useful.     

Milk Protein Polymorphism in Kangayam Cattle

This paper reports the milk protein polymorphism, the allele frequencies of variants and the possible linkages among various combinations of milk protein phenotypes in the Kangayam cattle of south India. Milk samples from 156 Kangayam cows were typed by starch gel and polyacrylamide gel electrophoresis for caseins and whey proteins, respectively. All the four milk protein components studied, _s1-casein, -casein, -lactoglobulin and -lactalbumin, exhibited polymorphism with high allele frequencies of 0.9231±0.0151 for _s1-casein C, 0.9263±0.0148 for -casein A, 0.9135±0.0159 for -lactoglobulin B and a relatively high frequency of 0.6218±0.0275 for -lactalbumin A. The mean heterozygosity estimated over all the four milk protein loci was 0.2420. Genetic equilibrium was observed among all the loci studied, except -lactalbumin. Linkage analysis confirmed the non-independence between _s1- and -caseins and between caseins and -lactalbumin phenotypes.     

Kappa-casein in milk. An immunochemical investigation of the kappa-casein content in milk from Danish cattle breeds

The ϰ-casein content in milk from the Danish cattle breeds SDM, RDM and Jersey was investigated by means of immunoquantitation in antibody containing agarose gels. The method, which determines the total ϰ-casein complex, is suitable for routine work. The reproducibility of the method was found to be ± 4.1 %.The ϰ-casein concentration, expressed as percentage of the total protein content in milk, was significantly lower in milk from SDM (21.5%) and RDM (21.3%) than in Jersey (23.9%). The investigation showed that the ϰ-casein constitutes a greater part of the protein in milk than previously assumed.The amount of ϰ-casein was positively correlated with the protein content in milk, but there was no relation between the relative ϰ-casein content and the percentage of fat, milk yield and the time after calving.Keyword: kappa-casein, milk, cattle, immunoquantitation     

High Prevalence of BlaCTX-M Group Genes in Aeromonas dhakensis Isolated from Aquaculture Fish Species in South Korea

The prevalence of resistant genes against β-lactams in 119 Aeromonas strains was determined. A large number (99.2%) of the present fish strains were resistant to one or more β- lactams including ceftiofur, amoxicillin-clavulanic acid, ampicillin, piperacillin and cefpodoxime. Among antibiotic resistance phenotypes, the simultaneous resistance to all β-lactams occurred in 25.2% (n=30) of all strains, which consisted of 18 strains of A. dhakensis, 8 strains of A. caviae, 2 strains of A. hydrophila and only one strain of A. veronii. For exploring genetic background of the antibiotic resistances, multiple PCR assays were subjected to detect β-lactamase-encoding genes, bla_TEM, bla_OXA-B and bla_CTX-M. In the results, the bla_TEM-1 gene was harbored in all strains, whereas only 3 strains harbored bla_OXA gene. In the case of bla_CTX-M gene, the gene was detected in 21.0% (25 out of 119) of all strains, which countered with 80% (20 out of 25) of A. dhakensis, 8% (2 out of 25) of A. caviae and 12% (3 out of 25) of A. hydrophila. In addition, most of the bla_CTX-M positive strains showed simultaneous resistance to all β-lactams (18 out of 30 strains). In sequence analysis for bla_CTX-M genes detected, they were CTX-M group 1-encoding genes including bla_CTX-M-33 from 3 eel strains of A. dhakensis. Therefore, A. dhakensis obtained from cultured fish could represent a reservoir for spreading genes encoding CTX-M group 1 enzymes and hence should be carefully monitored, especially for its potential risk to public health.     

Estrogen suppresses melatonin-enhanced hyperactivation of hamster spermatozoa

Hamster sperm hyperactivation is enhanced by progesterone, and this progesterone-enhanced hyperactivation is suppressed by 17β-estradiol (17βE₂) and γ-aminobutyric acid (GABA). Although it has been indicated that melatonin also enhances hyperactivation, it is unknown whether melatonin-enhanced hyperactivation is also suppressed by 17βE₂ and GABA. In the present study, melatonin-enhanced hyperactivation was significantly suppressed by 17βE₂ but not by GABA. Moreover, suppression of melatonin-enhanced hyperactivation by 17βE₂ occurred through non-genomic regulation via the estrogen receptor (ER). These results suggest that enhancement of hyperactivation is regulated by melatonin and 17βE₂ through non-genomic regulation.     

Regulation of matrix metalloproteinase-9 protein expression by 1α,25-(OH)2D3 during osteoclast differentiation

To investigate 1α,25-(OH)₂D₃ regulation of matrix metalloproteinase-9 (MMP-9) protein expression during osteoclast formation and differentiation, receptor activator of nuclear factor κB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) were administered to induce the differentiation of RAW264.7 cells into osteoclasts. The cells were incubated with different concentrations of 1α,25-(OH)₂D₃ during culturing, and cell proliferation was measured using the methylthiazol tetrazolium method. Osteoclast formation was confirmed using tartrate-resistant acid phosphatase (TRAP) staining and assessing bone lacunar resorption. MMP-9 protein expression levels were measured with Western blotting. We showed that 1α,25-(OH)₂D₃ inhibited RAW264.7 cell proliferation induced by RANKL and M-CSF, increased the numbers of TRAP-positive osteoclasts and their nuclei, enhanced osteoclast bone resorption, and promoted MMP-9 protein expression in a concentration-dependent manner. These findings indicate that 1α,25-(OH)₂D₃ administered at a physiological relevant concentration promoted osteoclast formation and could regulate osteoclast bone metabolism by increasing MMP-9 protein expression during osteoclast differentiation.     

Inhibition of transient receptor potential vanilloid type 1 through α2 adrenergic receptors at peripheral nerve terminals relieves pain

The activation of α₂ adrenergic receptors contributes to analgesia not only in the central nervous system but also in the peripheral nervous system. We reported that noradrenaline inhibits the activity of transient receptor potential vanilloid 1 (TRPV1) evoked by capsaicin through α₂ receptors in cultured rat dorsal root ganglion (DRG) neurons. However, it is unclear whether activation of TRPV1 expressed in peripheral nerve terminals is inhibited by α₂ receptors and whether this phenomenon contributes to analgesia. Therefore, we examined effects of clonidine, an α₂ receptor agonist, on several types of nociceptive behaviors, which may be caused by TRPV1 activity, and subtypes of α₂ receptors expressed with TRPV1 in primary sensory neurons in rats. Capsaicin injected into hind paws evoked nociceptive behaviors and clonidine preinjected into the same site inhibited capsaicin-evoked responses. This inhibition was not observed when clonidine was injected into the contralateral hind paws. Preinjection of clonidine into the plantar surface of ipsilateral, but not contralateral, hind paws reduced the sensitivity to heat stimuli. Clonidine partially reduced formalin-evoked responses when it was preinjected into ipsilateral hind paws. The expression level of α_2C receptor mRNA quantified by real-time PCR was highest followed by those of α_2A and α_2B receptors in DRGs. α_2A and α_2C receptor-like immunoreactivities were detected with TRPV1-like immunoreactivities in the same neurons. These results suggest that TRPV1 and α₂ receptors are coexpressed in peripheral nerve terminals and that the functional association between these two molecules causes analgesia.     

Metformin acts to suppress β-hydroxybutyric acid-mediated inflammatory responses through activation of AMPK signaling in bovine hepatocytes

The occurrence of bovine ketosis involves the accumulation of β-hydroxybutyric acid (BHBA), which contributes to the initiation and acceleration of hepatic metabolic stress and inflammation. Metformin has other beneficial effects apart from its medical intervention for diabetes, such as prevention of laminitis and hyper-triglyceridemic. AMPK maintains energy homeostasis and is the intracellular target of metformin action. This study aims to uncover the role of metformin in modulating BHBA-induced inflammatory responses through the activation of AMPK signaling. The hepatocytes were isolated from the liver tissue of mid-lactation multiparous Holstein cows (~160 d postpartum). Treatments were conducted as follows: treated with PBS for 18 h (control); pretreated with PBS for 12 h followed by treatment of 1.2 mM BHBA for 6 h (BHBA); pretreated with 1.5 mM or 3 mM metformin for 12 h followed by the BHBA treatment (1.2 mM) for 6 h (M(1.5)+B; M(3)+B). The inhibitor of AMPK, Compound C, at a concentration of 10 μM, was applied to substantiate the AMPK-dependent responses. RT-qPCR were applied for the mRNA expression while Western-blots and immunofluorescence were conducted for the target proteins expression. Among dose-dependent assays for BHBA, the concentration of BHBA at 1.2 mM activated NF-κB signaling by upregulating the expression of phosphorylated NF-κB and pro-inflammatory cytokines compared with the control cells (P < 0.05). Along with the upregulation of phosphorylated AMPKα and ACCα, metformin at 1.5 and 3 mM inactivated NF-κB signaling components (p65 and IκBα) and the inflammatory genes (TNFA, IL6, IL1B and COX-2) which were activated by BHBA. Additionally, BHBA inhibited cells staining intensity in EdU assay were increased by pretreatment with metformin. The activation of AMPK resulted in the increased gene and protein expression of SIRT1, along with the deacetylation of H3K9 and H3K14. However, the AMPK inhibitor compound C blocked this effect. Compared with BHBA treated cells, the protein expression of COX-2 and IL-1β were decreased by the pretreatment with metformin, and the inhibitory effect of metformin was released by compound C. The bound of NF-κB onto IL1B promoter displayed higher in BHBA group and this was suppressed by pretreatment with metformin (P < 0.05). Altogether, metformin attenuates the BHBA-induced inflammation through the inactivation of NF-κB as a target for AMPK/SIRT1 signaling in bovine hepatocytes.