The Influence of Cold-Pressed Linseed Cake Supplementation on Fatty-Acid Profile and Fat-Soluble Vitamins of Cows' Milk in an Organic Production System.

The aim of the study was to determine the effect of linseed cake supplementation during the winter period as a factor influencing the level of some bioactive components (milk composition, fatty-acid profile, and fat-soluble vitamins) in the milk fat fraction in cows kept on an organic farm. Forty multiparous (second and third lactation) Holstein-Friesian cows were selected that had 81 ± 12 days in milk and produced 15.08 ± 1.20 kg of milk/day. Two groups were created for the experiment: control (CTL; n = 20) and experimental (LC; n = 20). The experiment was divided into two periods: an initial period lasting 7 days in which the experimental group was habituated to the new supplement in their diet; the proper experimental phase, lasting 6 weeks, in which the cows in the experimental group received an individual daily dose of linseed cake (300 g/day/cow). Linseed cake supplementation had a positive impact on the levels of bioactive components (fatty-acid profile and fat-soluble vitamins) in the milk fat fraction. At the end of the trial, the concentration of C18:2 cis9 trans11, C18:1 trans11, α-retinol, α-tocopherol, and total antioxidant status increased 1.59-, 1.94-, 3.12-, 3.38-, and 3.09-fold, respectively, relative to the control levels. The use of linseed cake in winter on organic farms makes it possible to increase the antioxidant potential of milk, thereby eliminating the disparity in the quality of milk from the summer season compared to the winter season.

The Effect of Supplementation Using a Mixture of Fish Oil and Linseed on the Level of Immunomodulatory Components in Bovine Colostrum.

The aim of this study was to determine the effect of supplementing rations, with a mixture of fish oil and linseed, on the level of immunomodulatory components in colostrum. Twenty multiparous cows, that were three weeks before scheduled calving, had a body condition of 3-3.5, and had not been diagnosed with multiple pregnancies, were qualified for the experiment. The cows were divided into two groups: experimental (FOL) (n = 10) and control (CTL) (n = 10). The CTL group were individually given the standard food ration for dry cows for about 21 days before calving, while the FOL group received food rations that were enriched with 150 g of fish oil and 250 g of linseed (golden variety). Colostrum samples for testing were taken twice a day on the first and second days of lactation, and then once a day from the third to the fifth day of lactation. The experiment showed that the applied supplementation had an impact, in the form of increasing the fat, protein, IgG, IgA, IgM, vitamin A, C22:6 n-3 (DHA), and C18:2 cis9 trans11 (CLA) contents in colostrum; however, the C18: 2 n-6 (LA) and C20:4 n-6 (AA) contents decreased. Due to the lower quality of colostrum found in high-yield cows, and therefore in the Holstein-Friesian breed, it is possible to improve the quality by, among other things, introducing nutritional modifications during the second stage of the dry period.

Interaction between the level of immunoglobulins and number of somatic cells as a factor shaping the immunomodulating properties of colostrum.

The aim of this study was to investigate the association between immunoglobulins and SCC as a factor in shaping the content of the immunostimulatory components of colostrum. Seventy-eight multiparous Polish Holstein-Friesian cows were selected for the experiment. Colostrum samples were collected immediately after calving (up to a max. of 2 h). The cows were divided into groups according to the following levels: Immunoglobulins (IG class)-(IG1) over 50 g/L, (IG2) up to 50 g/L; SCC class-(SCC1) up to 400 000/ml, (SCC2) 400-800 000/ml, (SCC3) over 800 000/ml. Colostrum assigned to the IG1 SCC1 group had a statistically significant higher (p ≤ 0.01) concentration of both whey proteins and fatty acids compared to the IG1 SCC2 and SCC3 groups. The concentration of IgG, IgM, and IgA was shown to be higher in IG1 SCC1 than IG2 SCC3 by 226%, 149%, and 115%, respectively. The concentration of lactoferrin was shown to be higher in IG1 SCC1 than IG2 SCC3 by 149%. The determination of colostrum quality based on the concentration of immunoglobulins in the colostrum may not be sufficient because serum IgG concentrations at birth show a linear increase relative to colostrum SCC. A breakdown of colostrum into quality classes, taking into account the level of SCC, should therefore be introduced.

Different Patterns of mRNA Nuclear Retention during Meiotic Prophase in Larch Microsporocytes.

Recent studies show a crucial role of post-transcriptional processes in the regulation of gene expression. Our research has shown that mRNA retention in the nucleus plays a significant role in such regulation. We studied larch microsporocytes during meiotic prophase, characterized by pulsatile transcriptional activity. After each pulse, the transcriptional activity is silenced, but the transcripts synthesized at this time are not exported immediately to the cytoplasm but are retained in the cell nucleus and especially in Cajal bodies, where non-fully-spliced transcripts with retained introns are accumulated. Analysis of the transcriptome of these cells and detailed analysis of the nuclear retention and transport dynamics of several mRNAs revealed two main patterns of nuclear accumulation and transport. The majority of studied transcripts followed the first one, consisting of a more extended retention period and slow release to the cytoplasm. We have shown this in detail for the pre-mRNA and mRNA encoding RNA pol II subunit 10. In this pre-mRNA, a second (retained) intron is posttranscriptionally spliced at a precisely defined time. Fully mature mRNA is then released into the cytoplasm, where the RNA pol II complexes are produced. These proteins are necessary for transcription in the next pulse to occur.mRNAs encoding translation factors and SERRATE followed the second pattern, in which the retention period was shorter and transcripts were rapidly transferred to the cytoplasm. The presence of such a mechanism in various cell types from a diverse range of organisms suggests that it is an evolutionarily conserved mechanism of gene regulation.

The Effects of Host Plant Genotype and Environmental Conditions on Fungal Community Composition and Phosphorus Solubilization in Willow Short Rotation Coppice.

Phosphorus (P) is an essential plant nutrient. Low availability of P in soil is mainly caused by high content of Fe2O3 in the clay fraction that binds to P making it unavailable. Beneficial microbes, such as P solubilizing microorganisms can increase the available P in soil and improve plant growth and productivity. In this study, we evaluated the effects of environmental conditions (climate, soil parameters), plant genotype, and level of plant association (rhizosphere or endophytic root organism) on the abundance and diversity of phosphorus solubilizing microorganisms in a Salix production system. We hypothesized that a lower number of endophytic fungi may possess the ability to solubilize P compared to the number of rhizosphere fungi with the same ability. We also expect that the plant genotype and the experimental site with its environmental conditions will influence fungal diversity. Two Salix genotypes grown in pure and mixed cultures were investigated for their fungal microbiome community and diversity in the rhizosphere and endosphere during two growing seasons. We found that the rhizosphere fungal community was more diverse. A general dominance of Ascomycota (Dothideomycetes) and Basidiomycota (Tremellomycetes) was observed. The classes Agaricomycetes and Pezizomycetes were more frequent in the endosphere, while Tremellomycetes and Mortierellomycetes were more abundant in the rhizosphere. Plot-specific soil properties (pH, total organic carbon, and nitrogen) significantly influenced the fungal community structure. Among the culturable fungal diversities, 10 strains of phosphate solubilizing fungi (PSFs) from roots and 12 strains from rhizosphere soil were identified using selective media supplemented with di-calcium and tri-calcium phosphates. The fungal density and the number of PSF were much higher in the rhizosphere than in the endosphere. Penicillium was the dominant genus of PSF isolated from both sites; other less frequent genera of PSFs were Alternaria, Cladosporium, and Clonostachys. Overall the main factors controlling the fungal communities (endophytic vs. rhizosphere fungi) were the soil properties and level of plant association, while no significant influence of growing season was observed. Differences between Salix genotypes were observed for culturable fungal diversity, while in metagenomic data analysis, only the class Dothideomycetes showed a significant effect from the plant genotype.

Choosing source of microorganisms and processing technology for next generation beet bioinoculant.

The increase of human population and associated increasing demand for agricultural products lead to soil over-exploitation. Biofertilizers based on lyophilized plant material containing living plant growth-promoting microorganisms (PGPM) could be an alternative to conventional fertilizers that fits into sustainable agricultural technologies ideas. We aimed to: (1) assess the diversity of endophytic bacteria in sugar and sea beet roots and (2) determine the influence of osmoprotectants (trehalose and ectoine) addition during lyophilization on bacterial density, viability and salt tolerance. Microbiome diversity was assessed based on 16S rRNA amplicons sequencing, bacterial density and salt tolerance was evaluated in cultures, while bacterial viability was calculated by using fluorescence microscopy and flow cytometry. Here we show that plant genotype shapes its endophytic microbiome diversity and determines rhizosphere soil properties. Sea beet endophytic microbiome, consisting of genera characteristic for extreme environments, is more diverse and salt resistant than its crop relative. Supplementing osmoprotectants during root tissue lyophilization exerts a positive effect on bacterial community salt stress tolerance, viability and density. Trehalose improves the above-mentioned parameters more effectively than ectoine, moreover its use is economically advantageous, thus it may be used to formulate improved biofertilizers.

Use of somatic cell count as an indicator of colostrum quality.

The impact of cow mammary gland diseases on the quality of colostrum is not conclusively defined; research results are conflicting. However, it is widely believed that mastitis lowers the level of immunoglobulins and the quality of the colostrum produced. Therefore, the aim of this study was to determine the influence of somatic cell counts (SCC) on the colostrum immunostimulating and chemical components. The experiment was conducted on an experimental organic dairy farm in which a herd of approximately 250 cows was kept in a freestall housing system, with the average performance exceeding 6,000 kg of milk per lactation. Colostrum and milk samples were taken individually from each cow seven times during the experiment: from the first to second day after calving-twice per day, and from the third to fifth day after calving-once per day. Therefore, after preliminary analyses, the cows were divided into two groups based on the cytological quality of their colostrum at the first collection: 1. SCC ≤400,000 cells/ml (good quality colostrum; GCC- 18 cows), 2. SCC ≥ 400,000 cells/ml (low quality colostrum; LCC- 22 cows). The study found almost double the concentration of immunoglobulins and essential fatty acids in first milking colostrum in the GCC group than in colostrum from the LCC group. In addition, an increase in the concentration of lysozyme in first milking colostrum was associated with a decrease in the concentration of immunoglobulins. In addition, the increase in the level of lysozyme was associated with a decrease in the concentration of immunoglobulins. In conclusion, the SCC of first milking colostrum can be used as an indicator of colostrum quality.

Salt stress vs. salt shock - the case of sugar beet and its halophytic ancestor.

BACKGROUND: Sugar beet is a highly salt-tolerant crop. However, its ability to withstand high salinity is reduced compared to sea beet, a wild ancestor of all beet crops. The aim of this study was to investigate transcriptional patterns associated with physiological, cytological and biochemical mechanisms involved in salt response in these closely related subspecies. Salt acclimation strategies were assessed in plants subjected to either gradually increasing salt levels (salt-stress) or in excised leaves, exposed instantly to salinity (salt-shock). RESULT: The majority of DEGs was down-regulated under stress, which may lead to certain aspects of metabolism being reduced in this treatment, as exemplified by lowered transpiration and photosynthesis. This effect was more pronounced in sugar beet. Additionally, sugar beet, but not sea beet, growth was restricted. Silencing of genes encoding numerous transcription factors and signaling proteins was observed, concomitantly with the up-regulation of lipid transfer protein-encoding genes and those coding for NRTs. Bark storage protein genes were up-regulated in sugar beet to the level observed in unstressed sea beet. Osmotic adjustment, manifested by increased water and proline content, occurred in salt-shocked leaves of both genotypes, due to the concerted activation of genes encoding aquaporins, ion channels and osmoprotectants synthesizing enzymes. bHLH137 was the only TF-encoding gene induced by salt in a dose-dependent manner irrespective of the mode of salt treatment. Moreover, the incidence of bHLH-binding motives in promoter regions of salinity-regulated genes was significantly greater than in non-regulated ones. CONCLUSIONS: Maintaining homeostasis under salt stress requires deeper transcriptomic changes in the sugar beet than in the sea beet. In both genotypes salt shock elicits greater transcriptomic changes than stress and it results in greater number of up-regulated genes compared to the latter. NRTs and bark storage protein may play a yet undefined role in salt stress-acclimation in beet. bHLH is a putative regulator of salt response in beet leaves and a promising candidate for further studies.

Degrees of Antioxidant Protection: A 2-Year Study of the Bioactive Properties of Organic Milk in Poland.

The aim of this study was to determine the nutritional value of organic milk in Poland, investigate the influence of diet on antioxidant capacity and degree of antioxidant protection (DAP), and to examine the effect of season on the bioactive properties of milk from organic farms. From 2014 to 2015, 820 milk samples were collected from 6 organic farms during indoor feeding season (IDS) and pasture feeding season (PS). Pasture feeding season + corn grain (PSCG) cows' daily ration during pasture feeding season was enriched with 4 kg a day of corn to improve dietary energy balance. Milk obtained during PS was found to have a higher fat content, slight but significantly lower protein content compared with milk from IDS. The study showed that the content of monounsaturated fatty acids (MUFA) in milk fat was strongly linked to the concentration of polyunsaturated fatty acids (PUFA) and, to a lesser extent, on the supply of MUFA. The IDS data (concentration of vitamin E, A, and ß-carotene) showed the lowest values compared with the PS and PSCG groups. Total antioxidant status (TAS) and DAP showed an increasing trend in organic milk. PSCG was associated with highest level of DAP (9% higher than PS and 79% higher than IDS) and TAS (37% higher than PS and 79% higher than IDS). The results obtained show that supplementation of the basic ration with corn grain improved both TAS and DAP. The higher DAP and TAS value is responsible for product stability, considering the risk factor related to levels of cholesterol-oxide intake in humans.

Transcriptomic profiling of the salt stress response in excised leaves of the halophyte Beta vulgaris ssp. maritima.

Beta vulgaris ssp. maritima is a halophytic relative of cultivated beets. In the present work a transcriptome response to acute salt stress imposed to excised leaves of sea beet was investigated. Salt treatments consisted of adding NaCl directly to the transpiration stream by immersing the petioles of excised leaves into the salt solutions. Sequencing libraries were generated from leaves subjected to either moderate or strong salt stress. Control libraries were constructed from untreated leaves. Sequencing was performed using the Illumina MiSeq platform. We obtained 32970 unigenes by assembling the pooled reads from all the libraries with Trinity software. Screening the nr database returned 18,362 sequences with functional annotation. Using the reference transcriptome we identified 1,246 genes that were differentially expressed after 48 h of NaCl stress. Genes related to several cellular functions such as membrane transport, osmoprotection, molecular chaperoning, redox metabolism or protein synthesis were differentially expressed in response to salt stress. The response of sea beet leaves to salt treatments was marked out by transcriptomic up-regulation of genes related to photosynthetic carbon fixation, ribosome biogenesis, cell wall-building and cell wall expansion. Furthermore, several novel and undescribed transcripts were responsive to salinity in leaves of sea beet.

Effect of supplementation of cows diet with linseed and fish oil and different variants of ß-lactoglobulin on fatty acid composition and antioxidant capacity of milk.

BACKGROUND: The aim of this study was to determine the influence of polymorphic variants of ß-lactoglobulin in cows supplemented with linseed and fish oil on the fatty acid composition and antioxidant capacity of milk. From the herd of 320 Polish Holstein Friesian cows three groups of cows were selected according to the variants of ß-LG (ß-LGAA, ß-LGBB, ß-LGAB). During the first 7 days (the initial period) all the cows were fed the same total mixed ration (TMR) diet. From day 8 to 28,150 g fish oil and 250 g linseed (FOL) was added to the TMR diet of each cow. RESULTS: The results showed that the diet supplemented with FOL was effective in reducing atherogenic and thrombogenic indices. Introducing supplementation improved the antioxidant capacity: higher concentration of C18:2cis-9 trans-11, C20:5 n-3, C22:6 n-3, bioactive whey proteins and vitamin soluble in fat has been recorded. The results showed that ß-LGAA was associated with lower levels of atherogenic and thrombogenic indices and higher concentration of C22:5 n-6, phospholipids and ß-carotene. ß-LGBB favours a higher content of C18:1trans-11, C18:2cis-9 trans-11 and lactoferrin. ß-LGAB was associated with higher concentrations of C20:5 n-3, Lysozyme, α-retinol, α-tocopherol and total antioxidant status. CONCLUSION: Modification of the diet of cows with fish oil and linseed significantly influenced fatty acid composition and antioxidant properties of milk. The effect of ß-LG phenotype on the fatty acid composition and antioxidant capacity of milk is variable, which could partly be the result of a ß-LG phenotype × diet interaction.

Effect of fish oil and linseed supplementation on the protein composition of milk from cows with different ß-lactoglobulin phenotypes.

BACKGROUND: The objective of this study was to determine the impact of fish oil and linseed (FOL) supplements on the protein fraction levels of milk from cows with different phenotypes of ß-lactoglobulin. RESULTS: After 21 days of supplementation the study showed significantly higher concentrations of whey proteins, especially lysozyme (144% increase) and lactoferrin (45.5% increase), compared with milk from control cows (total mixed ration with no supplemented FOL). A reverse trend was demonstrated for casein, casein index and casein number (lower level). The most favourable change (higher level), in terms of lactoferrin, α-lactalbumin and bovine serum albumin contents in milk, was recorded in cows with the BB variant of ß-lactoglobulin. The highest level of lysozyme was recorded in the milk of cows with the AB variant of ß-lactoglobulin. CONCLUSION: The combined supplementation of fish oil and linseed had a positive impact on whey proteins in cow's milk. In addition, the phenotype of ß-lactoglobulin also played a role in milk protein composition. There is therefore a clear indication that nutritional experiments should take into account not only food supplements but also the genetic variants of ß-lactoglobulin.

The age of cows as a factor shaping the antioxidant level during a nutritional experiment with fish oil and linseed supplementation for increasing the antioxidant value of milk.

BACKGROUND: So far, in research studies, the age of cows has not been considered as a factor that may influence the changes in the content of milk ingredients with antioxidant properties modified by the feed supplementation. The aim of this study was to determine the influence of supplementation on the content of ingredients having antioxidant properties and to determine the influence of the age of cows taking part in the experiment on these changes. The experiment was conducted using 20 Polish Holstein Friesian cows, 10 primiparous and 10 multiparous. The combined supplementation of fish oil and linseed constituted the experimental factor. RESULTS: The milk of primiparous cows after 21 days of supplementation was characterised by a higher content of C18:1 trans-11, C18:2 cis-9, trans-11, α-retinol, α-tocopherol and ß-lactoglobulin compared to the milk of multiparous cows, in which a higher level of lactoferrin, C20:5 and ß-carotene was recorded. In both groups an increase in the total antioxidant status was noted (a higher level in the milk of primiparous cows). CONCLUSIONS: Modification of the diet of cows with fish oil and linseed significantly influenced antioxidant properties of their milk; however, the response of multiparous and primaparous cows was noticeably different to the supplement introduced.