Short communication: Relationships between α-lactalbumin and quality traits in bulk milk

The main objective of this study was to investigate whether the α-lactalbumin (α-LA) content of bulk milk is related with some known inflammatory markers and milk quality traits. An additional objective was to study whether combining α-LA, haptoglobin (Hp), and serum amyloid A (SAA) in an acute phase index (API) could be useful as an alternative marker for bulk milk quality. For the dairy industry, it is of great importance to receive high quality bulk milk for manufacture of liquid milk and dairy products. The somatic cell count (SCC) is currently used as an indirect marker for bulk milk quality, but because it is somewhat insensitive and unspecific, interest exists in alternative markers. Bulk milk samples were analyzed for α-LA, SCC, polymorphonuclear leukocyte count, Hp, SAA, fat, lactose, total protein and casein contents, casein number, protein composition, proteolysis, and coagulating properties. No significant differences were found in SCC, polymorphonuclear leukocyte count, Hp, or SAA between milk samples containing low, medium, or high concentrations of α-LA. Differences between α-LA groups were, however, found in some milk quality traits because high α-LA concentration was related to low concentrations of α_S1-, α_S2-, and β-caseins and high concentrations of lactose and β-lactoglobulin. A high API was related to low lactose content and casein number. Samples with high SCC contained less casein and had a lower casein number than milk with a low SCC, and proteolysis was significantly higher in high SCC milk than in low SCC milk. Neither α-LA nor API proved to be a better marker than SCC for the quality traits investigated, and α-LA was not considered to be a useful inflammatory marker in bulk milk.     

Short communication: Characterization of a monoclonal antibody for κ-casein B of cow's milk

A monoclonal antibody (antik-B) against an oligopeptide of 23 AA corresponding to the region 131-153 of bovine κ-casein (κ-CN) B was generated using the Human Combinatorial Antibody Library (HuCAL) technology. Both AA substitutions distinguishing κ-CN A and B are located in that region (positions 136 and 148). In this study, the reactivity of antik-B to milk samples collected from cows previously genotyped as CSN3*AA, CSN3*AB, and CSN3*BB was tested. According to Western blot results, antik-B recognized κ-CN B and it showed no cross-reactivity toward κ-CN A and other milk proteins. Furthermore, a modified Western blot method, urea-PAGE Western blot, was set up to assess the reactivity of antik-B toward all isoforms of κ-CN B. In conclusion, antik-B was specific to κ-CN B in milk and it seemed to be reactive toward all its isoforms.     

Short communication: The unusual genetic trend of αS1-casein in Alpine and Saanen breeds

Genetic variation at the α_S1-casein locus (CSN1S1) is recognized as being crucial in the selection of dairy goats for cheese yield. At this locus, the existence of alleles that have strong, intermediate, weak, and null favorable effects on cheese yield and curd firmness is well known. Selection for alleles that have a strong favorable effect has been deliberately carried out, especially in France. In fact, the importance of α_S1-casein in selection was recently confirmed in the selling policies of semen, where bucks are marketed according to their genotypes. We evaluated genotypes and alleles frequencies at the α_S1-casein locus in 491 Italian Saanen and Alpine goats and compared them with previous data to investigate their evolution over the past decade. We also estimated soft cheese yield in a subset of the most represented genotypes to quantify the economic importance of considering the genetic trend of α_S1-casein genotype frequencies. We found a significant increase in frequency of the allele with the strongest favorable effect, A (+12 and +13%), and of the intermediate allele E (+17 and +7%) in Saanen and Alpine goats, respectively. Surprisingly, the frequency of the strong allele B decreased strikingly over time (−12% in Saanen, −6% in Alpine from 2004 to 2012). This is consistent with the current marketing of semen, in that bucks that are homozygous for strong (AA and BB) and intermediate alleles (EE) and even heterozygous for these alleles (BE and AE) are considered equal. It is worth noting that this practice strongly penalizes the best breeders that have flocks composed almost entirely of goats that are homozygous for strong alleles. For heterozygous goats, we estimated an economic loss of €85 and €215 per goat per lactation, respectively, for AE and BE, compare with AA and BB genotypes. The marketing of buck semen should clearly differentiate these 2 alleles to ensure the best economic genetic progress at this locus.     

Short communication: Bovine κ-casein variants result in different angiotensin I converting enzyme (ACE) inhibitory peptides

Proteins in bovine milk are a common source of bioactive peptides. The peptides are released by the digestion of caseins and whey proteins. Peptides derived from the different genetic variants A, B, C, E, F1, F2, G1, G2, H, I, and J of bovine κ-casein (CSN3) were investigated for their inhibitory activities against angiotensin I converting enzyme (ACE). Amino acid sequences of the CSN3 variants were analyzed in silico to detect potential ACE inhibitory peptides. Besides known biologically active peptides, exclusive peptides were identified in some CSN3 variants and their biological activity was determined: within CSN3*B and CSN3*C, the ACE inhibitory peptide ASP (IC₅₀ = 242.3; the IC₅₀ value is equivalent to the micromolar concentration of peptide mediating a 50% inhibition of ACE activity) and within CSN3*C the peptide AHHP (IC₅₀ = 847.6) was detected. Furthermore, the peptides VSP (IC₅₀ = 21.8) and ACHP (IC₅₀ = 360.7) were identified in CSN3*F1 and CSN3*G2, respectively.     

Isolation and characterisation of κ-casein/whey protein particles from heated milk protein concentrate and role of κ-casein in whey protein aggregation

Milk protein concentrate (79% protein) reconstituted at 13.5% (w/v) protein was heated (90 °C, 25 min, pH 7.2) with or without added calcium chloride. After fractionation of the casein and whey protein aggregates by fast protein liquid chromatography, the heat stability (90 °C, up to 1 h) of the fractions (0.25%, w/v, protein) was assessed. The heat-induced aggregates were composed of whey protein and casein, in whey protein:casein ratios ranging from 1:0.5 to 1:9. The heat stability was positively correlated with the casein concentration in the samples. The samples containing the highest proportion of caseins were the most heat-stable, and close to 100% (w/w) of the aggregates were recovered post-heat treatment in the supernatant of such samples (centrifugation for 30 min at 10,000 × g). κ-Casein appeared to act as a chaperone controlling the aggregation of whey proteins, and this effect was stronger in the presence of α_S- and β-casein.     

Short communication: Oxidative status and incidence of mastitis relative to blood α-tocopherol concentrations in the postpartum period in dairy cows

Vitamin E supplementation, when combined with high blood α-tocopherol (>6.25 μg/mL) at dry off, has been reported to unexpectedly increased the risk for clinical mastitis in dairy cows. Furthermore, higher levels of oxidative stress in the postpartum period were related to higher risk of mastitis. The objective of the present study was to determine the relationship between various serum biomarkers of oxidative status, incidence of mastitis, and blood α-tocopherol concentrations at dry off and at calving. A total of 146 dairy cows from a commercial farm were used in an observational field study. All cows were supplemented with 3,000 and 50 IU/cow per day of all-rac-α-tocopherol during the dry period and lactation, respectively. Blood samples were collected at dry off and at calving. Serum was analyzed for α-tocopherol, levels of reactive oxygen metabolites (ROM), thiol groups (SH), and ferric-reducing ability. Three α-tocopherol groups at calving were created: high (>3 μg/mL), medium (2–3 μg/mL), and low (<2 μg/mL). Three α-tocopherol groups at dry off were created: high (>6.25 μg/mL), medium (4.25–6.25 μg/mL), and low (<4.25 μg/mL). All cases of clinical mastitis that occurred during the dry period and the entire subsequent lactation were verified by a veterinarian. No differences were observed in the incidence of mastitis between the 3 α-tocopherol groups based on the serum levels at dry off. Incidence of mastitis was 4 times lower in the high and medium groups when compared with the corresponding value for the low-α-tocopherol group based on the serum levels at calving. Lower levels of ROM and SH at dry off and at calving were found in the group of cows with the highest α-tocopherol values at dry off when compared with the corresponding values in the low-α-tocopherol group. The ROM values at dry off but not at calving were lower in the group of cows with the highest α-tocopherol values at calving when compared with the corresponding values in the low-α-tocopherol group. No differences were observed in ferric-reducing ability values between the 3 α-tocopherol groups at dry off or calving. No differences were observed in all biomarkers of oxidative status between healthy cows and those with mastitis. Thus, blood α-tocopherol is inversely related to certain biomarkers of oxidative stress in the postpartum period and incidence of mastitis. However, reduction in the incidence of mastitis is not mediated through a reduction in the levels of various biomarkers of oxidative stress.     

Short communication: Responses to increasing amounts of free α-linolenic acid infused into the duodenum of lactating dairy cows

Increasing the α-linolenic acid (LNA; 18:3 cis-9,cis-12,cis-15) content of milk fat might help promote consumers’ health. The objective of this study was to determine the potential to alter the content of LNA in milk by duodenal infusion of a free fatty acid mixture rich in LNA. Four multiparous lactating Chinese Holstein cows fitted with duodenal cannulas were administered 2 treatments in a crossover design: an LNA-rich fatty acid infusion at varying concentrations (0, 40, 80, 120, and 160 g/d) versus a basal infusate control. Dry matter intake was not affected by LNA infusions. Milk production tended to decrease and was quadratically affected as LNA infusion increased, but 4% fat-corrected milk yield was not changed. Milk fat content tended to increase linearly with LNA infusion. Milk protein content was not changed by LNA infusion, whereas milk lactose content and yield were decreased quadratically as LNA infusion increased. Increasing the amount of LNA infused into the duodenum linearly increased concentrations of 18:3 cis-9,cis-12,cis-15 (0.61 to 25.4 g/100 g of total fatty acids) and 18:2 cis-9,cis-12 in milk fat. Increasing LNA decreased the percentages of 4:0, 14:0, and 16:0 fatty acids linearly. Increasing LNA also linearly decreased the percentages of 18:1 cis-9 and 18:2 cis-9,trans-11 in milk fat. Milk fat content of 20:5 cis-5,cis-8,cis-11,cis-14,cis-17 was quadratically affected, whereas concentrations of 18:0, 18:1 trans-9, 18:1 trans-11, and 18:2 trans-10,cis-12 were not affected. Increasing the supply of 18:3 cis-9,cis-12,cis-15 to the small intestine linearly increased 18:3 cis-9,cis-12,cis-15 in milk fat and markedly altered milk fat composition.     

Short communication: Molecular genetic characterization of ovine αS1-casein allele H caused by alternative splicing

Sequencing of ovine CSN1S1*H cDNA showed an absence of exon 8 in comparison with GenBank sequences; the absence was confirmed by protein sequencing. We demonstrated that this allelic aberration is the result of a deletion of 4 nucleotides, the last 3 of exon 8 and the first 1 of intron 8, which are replaced by an insertion of 13 nucleotides in the DNA sequence. The insertion is a precise duplication of a part of the adjacent intronic sequence of CSN1S1*C″. These sequence differences result in an inactivation of the splice donor sequence distal to exon 8, leading to upstream exon skipping during the serial splice reactions of the ovine CSN1S1*H pre-mRNA, and may affect the specific casein expression as well as protein characteristics.     

Short communication: Effect of αS1-casein (CSN1S1) and κ-casein (CSN3) genotypes on milk composition in Murciano-Granadina goats

The effects of the caprine α_S1-casein (CSN1S1) polymorphisms on milk quality have been widely demonstrated. However, much less is known about the consequences of the κ-casein (CSN3) genotype on milk composition in goats. Moreover, the occurrence of interactions between CSN3 and CSN1S1 genotypes has not been investigated. In this study, an association analysis between CSN1S1 and CSN3 genotypes and milk quality traits was performed in 89 Murciano-Granadina goats. Total milk yield as well as total protein, fat, solids-not-fat, lactose, α_S1-casein (CSN1S1), and α_S2-casein (CSN1S2) contents were recorded every other month during a whole lactation (316 observations). Data analysis using a linear mixed model for repeated observations revealed no interaction between the CSN1S1 and CSN3 genotypes. With regard to the effect of the CSN3 locus, AB and BB genotypes were significantly associated with higher levels of total casein and protein content compared with the AA CSN3 genotype. In strong contrast with French breeds, the CSN1S1 genotype did not affect protein, casein, and fat concentrations in Murciano-Granadina goats. These results highlight the importance of taking into consideration the CSN3 genotype when performing selection for milk composition in dairy goats.     

Short communication: In vitro ruminal fermentability of a modified corn cultivar expressing a thermotolerant α-amylase

The fermentability of a corn cultivar that expresses a thermostable α-amylase (CA3272) was evaluated under various in vitro conditions. The CA3272 corn was developed as a replacement to microbial enzyme additions during the high-temperature processing of corn to produce ethanol. The α-amylase activity in the corn might have the potential for positive effects on ruminant performance if incorporated into the ration. Four corn cultivars were evaluated in an in vitro ruminal fermentation where the digestion of starch was measured after 6 h. The cultivars included a flint corn, an opaque corn, CA3272, and its near-isogenic counterpart (IC). The flint corn produced less total volatile fatty acids (18.4 mM) than the other 3 corns (average of 25.3 mM), supporting the fact that it had the highest concentration of prolamins, which are negatively associated with starch availability. A second 6-h in vitro ruminal fermentation evaluated mixtures of the CA3272 and IC corns (0, 25, 50, 75, and 100% concentrations of CA3272). Total volatile fatty acid production was not different among treatments for any proportions of CA3272. In a third in vitro experiment, there was a small but significant difference in starch degradation of CA3272 compared with IC (90.6 vs. 89.7%) but this difference is most likely not biologically relevant. In a fourth in vitro experiment, CA3272 and IC were incubated in water at 40 and 65°C for 24 h. Degradation of starch from native amylase activity at 40°C was 1.99 and 1.60% for CA3272 and IC, respectively, but when they were incubated at 65°C, starch degradation was 10.56 and 0.85% for CA3272 and IC, respectively. These data demonstrate that amylase activity in CA3272 is expressed at a high temperature (65°C) but at the physiological temperature expected in a rumen of a cow (39-40°C), expression of amylase activity does not appear to be sufficient to have any positive (or negative) effects on ruminal metabolism.     

The β-lactoglobulin content of bovine milk: Development and application of a biosensor immunoassay

An optical biosensor immunoassay exploiting surface plasmon resonance is described for the quantification of β-lactoglobulin in milk. Samples were diluted with buffer, and the protein estimated from binding with a polyclonal antibody immobilised on the sensor surface. Analytical method performance characteristics including range, detection limit, precision and accuracy were determined and reported. The temporal variability in the β-lactoglobulin content of milk from pasture-fed cows during early lactation and across a production season was investigated. The content of β-lactoglobulin decreased from >10 mg mL⁻¹ in early colostrum to <5 mg mL⁻¹ in mature milk, and the β-lactoglobulin content of skim milk powder trended from 25 to 60 mg g⁻¹ across a season. In view of its allergenicity, these data will improve understanding of the expression of innate β-lactoglobulin in the milk of pasture-grazed dairy herds, thereby providing information that is applicable to the formulation of bovine milk-based products.     

Goat milk allergenicity as a function of αs₁-casein genetic polymorphism

Cow milk allergy is the most frequent allergy in the first years of life. Milk from other mammalian species has been suggested as a possible nutritional alternative to cow milk, but in several cases, the clinical studies showed a high risk of cross-reactivity with cow milk. In the goat species, α_S1-casein (α_S1-CN), coded by the CSN1S1 gene, is characterized by extensive qualitative and quantitative polymorphisms. Some alleles are associated with null (i.e., CSN1S1*0₁) or reduced (i.e., CSN1S1*F) expression of the specific protein. The aim of this work was to obtain new information on goat milk and to evaluate its suitability for allergic subjects, depending on the genetic variation at α_s1-CN. Individual milk samples from 25 goats with different CSN1S1 genotypes were analyzed by sodium dodecyl sulfate PAGE and immunoblotting, using monoclonal antibodies specific for bovine α-CN and sera from children allergic to cow milk. A lower reaction was observed to 2 goat milk samples characterized by the CSN1S1* 0₁0₁ and 0₁F genotypes. Moreover, a fresh food skin prick test, carried out on 6 allergic children, showed the lack of positive reaction to the 0₁0₁ milk sample and only one weak reactivity to the 0₁F sample. The risk of cross-reactivity between cow and goat milk proteins suggests the need for caution before using goat milk for infant formulas. However, we hypothesize that it can be used successfully in the preparation of modified formulas for selected groups of allergic patients. The importance of taking the individual goat CN genetic variation into account in further experimental studies is evident from the results of the present work.     

Effects of β-κ-casein (CSN2-CSN3) haplotypes and β-lactoglobulin (BLG) genotypes on milk production traits and detailed protein composition of individual milk of Simmental cows

The aim of this study was to investigate the effects of CSN2-CSN3 (β-κ-casein) haplotypes and BLG (β-lactoglobulin) genotypes on milk production traits, content of protein fractions, and detailed protein composition of individual milk of Simmental cows. Content of the major protein fractions was measured by reversed-phase HPLC in individual milk samples of 2,167 cows. Protein composition was measured as percentage of each casein (CN) fraction to total CN and as percentage of β-lactoglobulin (β-LG) to total whey protein. Genotypes at CSN2, CSN3, and BLG were ascertained by reversed-phase HPLC, and CSN2-CSN3 haplotype probabilities were estimated for each cow. Traits were analyzed by using a linear model including the fixed effects of herd-test-day, parity, days in milk, and somatic cell score class, linear regressions on haplotype probabilities, class of BLG genotype, and the random effect of the sire of the cow. Effects of haplotypes and BLG genotypes on yields were weak or trivial. Genotype BB at BLG and haplotypes carrying CSN2 B and CSN3 B were associated with increased CN content and CN number. Haplotypes including CSN3 B were associated with increased κ-CN content and percentage of κ-CN to total CN and with decreased percentages of α_S1- and γ-CN to total CN. Allele CSN2 B had the effect of increasing β-CN content and decreasing content of α_S1-CN. Haplotypes including allele CSN2 A¹ exhibited decreased β-, α_S2-, and γ-CN concentrations and increased α_S1- and κ-CN contents, whereas CSN2 I had positive effects on β-CN concentration and trivial effects on content of other protein fractions. Effects of haplotypes on CN composition were similar to those exerted on content of CN fractions. Allele BLG A was associated with increased β-LG concentration and percentage of β-LG to total whey protein and with decreased content of other milk proteins, namely β-CN and α_S1-CN. Estimated additive genetic variance for investigated traits ranged from 14 to 39% of total variance. Increasing the frequency of specific genotypes or haplotypes by selective breeding might be an effective way to change milk protein composition.     

Immunoreactive properties of α-casein and κ-casein: Ex vivo and in vivo studies

The aim of this study was to evaluate the ex vivo and in vivo studies immune potential of α- and κ-casein. Ex vivo, naïve mouse splenocytes were stimulated with α- or κ-casein. After 120 h of culture, the proliferation index (PI), determined by 3-(4,5 dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and carboxyfluorescein diacetate N-succinimidyl ester (CFSE) staining, did not vary for either antigen, suggesting similar ex vivo immunogenic potential of both casein fractions. In vivo, BALB/ccmdb mice were sensitized with α- or κ-casein and then gavaged with primary antigen. Mice immunized with α-casein had higher levels of IgG (2^16.33) and IgA (2^10.22) in serum at the end of the experiment compared with mice immunized with κ-casein (2¹⁵ and 2^9.3 for IgG and IgA, respectively). The use of α-casein for mouse immunization and ex vivo lymphocyte stimulation resulted in higher concentrations of secreted cytokines (IL-4, IL-10) compared with κ-casein stimulation. This is consistent with increasing regulatory T cell (Treg) lymphocyte populations, independent of the antigen used for stimulation. In summary, the immunogenic potential of α- and κ-casein was similar. Humoral and cellular immune responses confirmed their strong, independent potential to induce B and T cells. We propose that the lymphocyte proliferation index be used as an initial screening for protein immunogenicity.     

Effect of κ-casein B relative content in bulk milk κ-casein on Montasio, Asiago, and Caciotta cheese yield using milk of similar protein composition

The aim of this study was to investigate the effect exerted by the relative content of κ-casein (κ-CN) B in bulk milk κ-CN on coagulation properties and cheese yield of 3 Italian cheese varieties (Montasio, Asiago, and Caciotta). Twenty-four cheese-making experiments were carried out in 2 industrial and 1 small-scale dairy plant. Detailed protein composition of bulk milk of 380 herds providing milk to these dairies was analyzed by reversed-phase HPLC. To obtain 2 experimental milks differing in the relative content of κ-CN B in κ-CN, herds were selected on the basis of bulk milk protein composition and relative content of κ-CN genetic variants. Milk was collected and processed separately for the 2 groups of selected herds. A difference of 20% in the relative content of κ-CN B in κ-CN was obtained for the 2 experimental milks for Montasio and a difference of 15% for Asiago and Caciotta. The 2 experimental milks were of similar protein and CN content, casein number, pH, CN composition, and β-CN genetic composition. For each cheese-making trial, amounts of milk, ranging from 2,000 to 6,000 kg, were manufactured. Each vat contained milk collected at least from 4 dairy herds. Cheese yield after brining and at the end of the aging was recorded. Milk with a greater proportion of κ-CN B in κ-CN (HIGHB) exhibited similar coagulation properties and greater cheese yield compared with milk with a lower proportion of κ-CN B in κ-CN (LOWB). The increased cheese yield observed for HIGHB when manufacturing Montasio cheese was ascribed to a greater fat content compared with LOWB. The probability of HIGHB giving a cheese yield 5% greater than that of LOWB ranged from 51 to 67% for Montasio cheese, but was less than 21% for Asiago and Caciotta cheeses. Variation in relative content of κ-CN B in κ-CN content did not relevantly affect industrial cheese yield when milks of similar CN composition were processed. An indirect effect due to the increased κ-CN content of κ-CN B milk is thought to explain the favorable effects of κ-CN B on cheese yield reported in the literature.     

Occurrence of β-casein fragments in cold-stored and curdled river buffalo (Bubalus bubalis L.) milk

The safeguard of river buffalo Mozzarella cheese, a Protected Designation of Origin dairy product, has prompted an analytical study to trace the milk and curd used as raw material in cheesemaking. This is to prevent the illegal use of milk or curd from different geographical areas outside of those indicated in the official production protocol. For this purpose, we studied primary proteolysis occurring in fresh and frozen milk and curd to identify a molecular marker that could indicate the raw material used. Whole casein from frozen river buffalo milk was separated using cation-exchange chromatography and sodium dodecyl sulfate-PAGE, and a protein component with an estimated molecular weight of 15.3 kDa was detected. This protein component was revealed in fresh river buffalo milk as a faint electrophoresis band, which drastically increased in intensity in refrigerated and frozen milk as well as in curd and was found to be associated with β-CN through hydrophobic interaction. By using matrix-assisted laser desorption/ionization-time of flight peptide mass mapping, this component was identified as the C-terminal fragment f(69-209) of β-CN (expected molecular weight of 15,748.8 Da). β-Casein f(69-209), originating from the early hydrolysis of Lys₆₈-Ser₆₉ by plasmin, has no counterpart in bovine milk. The increased rate of hydrolysis by plasmin toward the cleavage site Lys₆₈-Ser₆₉ has to be ascribed to the elevated proline content of the peptide 61-73. The favored production of β-CN f(69-209) has also drawn attention to the complementary proteose peptone β-CN f(1-68) that is presumed to play a physiological role in inducing milk secretion similar to that of β-CN f(1-29). The higher in vivo and in vitro production rate, compared with γ₁-CN formation, indicates that β-CN f(69-209) and its complementary fragment are candidate molecular markers to evaluate milk and curd freshness. We used indirect ELISA analysis based on the determination of remaining nonhydrolyzed β-CN to perform a quantitative evaluation of proteolysis.     

Short communication: Born to be a loser cow?

Over the last few years, an increasing awareness has arisen in Denmark of the existence of cows with a generally lowered health and production status, referred to as “loser cows.” A previous study has estimated that the overall prevalence of loser cows in Danish Holstein herds is 3.2%. The aim of this study was to estimate genetic and phenotypic parameters for the loser cow state and the underlying traits: lameness, hock lesions, other cutaneous lesions, and condition of hair coat. Records on 6,098 cows were analyzed with an animal model including fixed effects of herd, season of scoring and location of scoring, age at first calving, lactation stage, and parity in addition to additive genetic effects and permanent environmental effects. The heritability of the loser cow score was 0.08 and for the underlying traits the heritability ranged from 0.05 to 0.12. The genetic correlations between various pairs of traits included in the loser cow score ranged from 0.04 to 0.68 and the phenotypic correlations ranged from 0.09 to 0.21. The genetic and phenotypic correlations between the loser cow score and the underlying traits ranged from 0.25 to 0.89 and 0.20 to 0.85, respectively, supporting the concept of the loser cow score. The traits included in the loser cow score are easy to assess and all showed genetic variation. They are therefore suitable for inclusion in a total merit index aimed at breeding for more robust cows.     

Flavan-3-ol contents,anti-oxidative and α-glucosidase inhibitory activities of Cynomorium songaricum

Flavan-3-ol oligomers from the stems of Cynomorium songaricum were found to show potent SOD-like activity with one of the dimers being most potent. The flavan-3-ols also showed inhibitory activity on α-glucosidase, implying the beneficial effects of this herb on diabetes patients. The total (extractable and non-extractable) tannin content of this herb was found to be as high as 18.3%. The contents of catechin, flavan-3-ol oligomers and extractable tannins using 70% acetone, methanol or water were analysed. It was found that 70% acetone was the most efficient extract solvent amongst these three solvents, especially for higher flavan-3-ol oligomers.