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Abstract. Lactase-phlorizin hydrolase (LCT), more commonly known as lactase, is an enzyme responsible for cleaving lactose into absorbable monosaccharides, glucose and galactose. LCT deficiency (hypolactasia - HL) is caused by a decreased activity of LCT in the small intestinal villi and potentially results in lactose malabsorption what may lead to the development of clinical symptoms (diarrhea, bloating, flatulence and cramps) and avoiding milk products in the diet. HL is the world's most common enzyme deficiency in humans. HL exists in three distinct forms - congenital, primary and secondary. Adult type hypolactasia (ATH) is the most common phenotype found in human. It is a genetically predetermined physiological condition inherited through an autosomal recessive mode which results in a decline of lactase activity after weaning. ATH is associated with the LCT -13910 C>T polymorphism worldwide, except in Africa. Lactase non-persistence has been observed in individuals with the C/C-13910 genotype, whereas lactase persistence in subjects with remaining allelic variants. Small intestine biopsy is the only diagnostic procedure allowing for the direct measurement of LCT activity, however due to its invasive nature it is hardly accepted by patients. Therefore, LCT status is often inferred simply by assessing the patient's lactose digestion. A lactose tolerance test can be performed after lactose load and then measuring blood glucose concentration or breath hydrogen (preferably hydrogen and methane) expiration. A genetic test of the C/T-13910 polymorphism is also available at present. It is a reliable method in excluding/confirming ATH predisposition. However, it definitely does not assess lactose tolerance or malabsorption.
The objective of this study was to estimate genetic correlations of lactose percentage and urea concentration in milk with conformation traits related to udder and legs of Polish Holstein-Friesian cows. Data consisted of 5,813 test-day records and type scores of 791 primiparous cows. The analysis involved two descriptive traits (udder, feet and legs, scored from 50 to 100) and 11 linearly scored traits (describing udder: fore udder height, rear udder height, central ligament, udder depth, udder width, fore teat placement, teat length, rear teat placement; describing legs: rear legs - side view, foot angle, rear legs - rear view; on a scale of 1 to 9). Genetic correlations were calculated based on (co)variances estimated using the Bayesian method via Gibbs sampling and the multitrait animal model. Genetic correlations between lactose content and conformation traits ranged from -0.18 to 0.23, while those between milk urea concentration and conformation traits ranged between -0.02 and 0.43, respectively. Absolute values of average genetic correlations with daily lactose percentage exceeded 0.15 only for udder (descriptive trait) and several linearly scored traits, i.e. central ligament, udder depth, rear teat placement, and rear legs - rear view. Milk urea content was weakly or moderately genetically correlated with six type traits: udder, and five linearly scored traits: fore udder height, central ligament, udder width, teat length, and rear legs - side view. Absolute values of genetic correlations between these traits exceeded 0.15. Our results showed that type traits connected with udder were more highly genetically correlated with both lactose and milk urea contents than type traits describing legs. It meant that an increase in both lactose percentage and urea concentration in milk might be expected as an indirect response to selection for better udder, whereas selection for improvement of legs would not affect lactose percentage and milk urea content.
In the future an approach incorporating cows’ measured phenotypes and marker genotypes of cows and bulls within a single model can be applied. The most important advantage of such a model is the simultaneous use of pedigree and marker-based genomic relationship data. Such a solution allows the use of both genotyped and non-genotyped animals in the prediction procedure. This pilot study is aimed towards implementation of a one-step approach in a random regression test day model context for the Polish Holstein Friesian population, considering various ways of adjusting the relationship matrix. Data consisted of 890 animals (10 genotyped bulls, 100 cows with phenotypic data and 780 ancestors without genotypes or phenotypes). Random regression test day models with a polygenic effect on milk yield modeled by second order Legendre polynomials for the estimation of variance-covariance parameters and were used for prediction of genomically enhanced breeding values (GEBV). In this model, a matrix combining pedigree and marker-based information was used instead of a traditional numerator relationship matrix. In this matrix the proportions of information coming from pedigree and markers were defined by weighting parameters w and 1-w for pedigree and marker-based information matrices, respectively. Various weights of the two sources of information were considered. The accuracy of GEBV both for genotyped bulls and for cows with phenotypes was highest for weighting parameter w=0 and lowest for w=l. Incorporating genomic information into a conventional genetic evaluation improves reliabilities of breeding value prediction, however, pedigree information is important to maintain the stability of evaluation for non-genotyped animals. Implementation of the single-step approach in a random regression test day model framework is very attractive for genomic prediction in dairy cattle, since it allows to incorporate genomic information directly into a conventional genetic evaluation. However, for accurate predictions it is essential to achieve the right balance between the numerator relationship and markers-based relationship information.
The non-enzymatic reaction between reducing sugars and proteins, known as glycation, has received increased attention from nutritional and medical research. In addition, there is a large interest in obtaining glycoconjugates of pure well-characterized oligosaccharides for biological research. In this study, glycation of bovine serum albumin (BSA) by d-glucose, d-galactose and d-lactose under dry-heat at 60°C for 30, 60, 120, 180 or 240 min was assessed and the glycated products studied in order to establish their biological recognition by lectins. BSA glycation was monitored using gel electrophoresis, determination of available amino groups and lectin binding assays. The BSA molecular mass increase and glycation sites were investigated by mass spectrometry and through digestion with trypsin and chymotrypsin. Depending on time and type of sugar, differences in BSA conjugation were achieved. Modified BSA revealed reduction of amino groups’ availability and slower migration through SDS/PAGE. d-Galactose was more reactive than d-glucose or d-lactose, leading to the coupling of 10, 3 and 1 sugar residues, respectively, after 120 minutes of reaction. BSA lysines (K) were the preferred modified amino acids; both K256 and K420 appeared the most available for conjugation. Only BSA-lactose showed biological recognition by specific lectins.
Neutralised whey obtained after lactic fermentation and acidic coagulation of milk proteins carried out during tvarog (white cheese) production was concentrated selectively 3 times by means of membrane nanofiltration. The aim of this research was to identifiy properties obtained in this way in the industrial liquid whey preparation of milk components (WPMC) and utilisation of its natural water phase as a component of wheat dough. The main principles were that the use of the whey preparation will limit the amount of the technological water as a result of inclusion of this whey water phase and that the elimination or reduction of salt added during wheat dough formation will be possible. It was established that the acceptance of a slight salty taste in bakery products (rolls or bread) is possible by the addition of WPMC using both types of wheat flour (type 500 or type 750) except for the technological salt in the dough formation process. Wheat products with WPMC had a tendency to form the crumb characterised with small porosity and diminished volume. It was clearly noticed that incorporation of lactose doubled the content of disaccharides in crumb of experimental products. That phenomenon affords the opportunity for lactose supply as a substrate for the intestinal microflora both for children and adults. The participation of WPMC preparation, with the exception of technological salt, influenced changes in the proportion of individual elements, especially the proportion between sodium and potassium which accounted for 1:4, compared to the control sample where it was 3:1. The content of calcium and magnesium, whose active transport in a human body may proceed with hydrolysed lactose, could be increased significantly. This afforded the possibility for obtaining low-sodium products, being attractive special or prophylaxis food to be used in obesity, heart diseases, hypertension and as the support of the intestinal ecosystem.
Ethanol fermentation of lactose mash by biocatalyzers immobilized in calcium alginate was studied in order to improve the process productivity and economy. The fermentation effectiveness of S. cerevisiae co-immobilized with a ß-galactosidase and the directly lactose-fermenting immobilized yeast: K. fragilis and C. pseudotropicalis were compared. The application of the immobilized K. fragilis produced desirable results and even after its 10th (20 days) fermentation the immobilized yeast provided the stable high fermentation level (on average about 6%/vol of ethanol) while maintaining its activity. Such lactose-mash fermentation was greater than in conventional method (by free cells). In addition, the application of S. cerevisiae co-immobilized with ß-galactosidase produced somewhat greater fermentation levels than the conventional method.
Continuous cultivation of Trichodcnna reesei M-7 mutant was performed at different temperatures (26, 30 and 34°C) with 1% lactose or glucose alone or a total of 1% mixtures of both sugars at different ratio. The secretion of individual enzymes was affected by the ratio of cellulase inducer (lactose) to the repressor (glucose). The ratio of enzyme secretions was additionally modified by the temperature of cultivations. An insignificant increase of nonspecific activity of cellulases (FPU) and significant increase of aryl-ß-glucosidase activity were observed at 26°C with lactose/glucose ratio of 3:1 and 1:1. However, when cultivated both at 30 or 34°C, the cellulolytic activities (FPU) increased significantly in the presence of 1% glucose alone. The activity of xylanases increased significantly (about 8-fold) with lactose/glucose ratio of 1:1 at 30°C. The increased synthesis of lytic enzymes (proteases, ß-1,3-glucanases) correlated with increased glucose concentration in the feeding medium and this effect potentiated at 34°C of cultivation.
Transgalactosylation properties of the commercial preparations of beta-galactosidase: Lactase and Maxilact and of the preparation synthesized by Penicillium canescens, were compared.The effect of 2% glucose addition to 20% milk permeate solution on the process of formation of galac- tooligosaccharides during the enzymatic bioconversion of lactose by the examined beta-galactosidase preparations, was also studied. It was found that enzymatic synthesis of galactooligosaccharides in the process of lactose bioconversion was greatly dependent on beta-galactosidase used and that the presence of glucose in lactose solution already at the beginning of hydrolysis was favourable for the formation of oligosaccharides.
Cultivation has been performed of isolated earlier low-protease mutants and parental strain in the presence of novel inducers of cellulases production - lactulose and lactobionic acid mixed with lactose in the ratio 1:1. Nine among ten tested during batch cultivation low-protease mutants exhibited lower ability for the production of proteases than parental strain M-7. Higher enzymatic activities have been estimated (cellulolytic, xylanolytic and beta - galaktosidase) of culture filtrates during cultivation in the presence of mixtures of lactobionic acid and lactose in comparison to lactulose and lactose. Tested, during continuous cultivation, low-protease mutant T. reesei Mp5 exhibited high resistance for the temperature shifting. FPU activities of culture filtrates were stable in the high range of temperature of cultivation (26-34°C). In addition current knowledge about correlation between protease and other enzymes of culture fluids of selected filamentous fungi has been reviewed. The ways of preventing homologous and heterologous protein produced by fungi and bacteria from enzymatic cleavage by proteases have been also described. This work is an introduction for future studies on correlation between proteases and other enzymes in culture filtrates of Trichoderma reesei.
A feeding trial was performed on 1400 Big-6 turkey toms divided into experimental groups subject to the use of dietary supplements. The ain of this study was to evaluate the efficacy of the probiotic supplement Bactocell, containing lactic acid bacteria Pediococcus acidilactici, and lactose, administered to turkeys separately or in combination, as well as a formic acid supplement. The addition of the probiotic under test (lactic acid bacteria Pediococcus acidilactici) to diets for turkeys contributed to higher daily gains and lower feed consumption per kg weigh gain only during the first 12 weeks of their life. Diet supplementation with lactic acid bacteria and lactose reduced mortality rates. A slaughter value analysis revealed only a slightly (by approximately 1%) higher content of breast muscle and a lower content of thigh muscle in birds fed diets supplemented with lactic acid bacteria. Turkeys receiving lactic acid bacteria or lactose and a combination of both these supplements were characterized by a higher fat content of meat and slightly lower pH values, whereas meat from turkeys fed lactose-supplemented diets was darker in color. The addition of formic acid Acidum formicum to diets for turkeys contributed only to lower mortality rates.
 Recently, glyco-therapy is proposed to prevent the interaction of bacterial lectins with host ligands (glycoconjugates). This interaction represents the first step in infection. Neoglycans referred to as PSA-Lac (PSA-Glu (β1-4) Gal) were obtained by conjugation of porcine serum albumin (PSA) with lactose at 80 °C, 100 °C and 120 ºC. Characterization studies of the products showed that PSA could contain 1, 38 or 41 added lactoses, depending on the reaction temperature. These neoglycans were approximately 10 times more glycated than PSA-Lac obtained in previous work. Lactose conjugation occurred only at lysines and PSA-Lac contained terminal galactoses as confirmed by Ricinus communis lectin recognition. Furthermore, Escherichia coli K88+, K88ab, K88ac and K88ad adhesins showed affinity toward all PSA-Lac neoglycans, and the most effective was the PSA-Lac obtained after 100 ºC treatment. In vitro, this neoglycan partially inhibited the adhesion of E. coli K88+ to piglet mucin (its natural ligand). These results provide support for the hypothesis that glycated proteins can be used as an alternative for bioactive compounds for disease prevention.
The aim of this paper was to investigate the impact of clinical lameness in Polish Holstein-Friesian dairy cows on milk yield as well as on fat, protein, and lactose yields during early lactation (120 d). The dataset includes 312 monthly test-day milk yields and milk composition records from 78 cows, which calved from October 2008 to April 2009. Twice a week, during the first 120 d postpartum, the cows were examined for lameness and they were scored fortnightly according to a 5-point locomotion scoring system. Factors affecting milk yield and milk components included calving season, parity, month of lactation, and degree of lameness. In cows, which were clinically lame for a month and which were clinically lame for more than a month (maximum per 2 months), the total mean reduction in milk yield per 120-d lactation was approximately 308 and 283 kg, respectively, as compared with cows, which were never lame in early lactation.
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