Research Institute for Cattle Breeding (Czechia)

The experiment was designed to evaluate the influence of dietary supplementation with L-Citrulline (L-Cit) on reproductive outcomes, serum reproductive hormone profiles, ruminal and intestinal microbial communities, serum metabolites, and their metabolic associations in Simmental cows, thereby providing a theoretical basis for its application in ruminant production. All cows were screened by vaginal mucus microscopy (no pathogenic bacteria detected) and B-ultrasound examination (no ovarian cysts or uterine inflammation observed) to confirm the absence of reproductive disorders. A total of 240 multiparous Simmental cows, 3–4 years of age with an average body weight of 470 ± 15 kg, were randomly allocated into three groups of 80 animals each: a control group, Group I, and Group II. The control group received only the basal diet, whereas Group I and Group II were provided with the basal diet supplemented with 7 g/d and 14 g/d of L-Cit per cow, respectively. Day 0 of the trial was defined by the first intramuscular injection of prostaglandin (PG, 0.4 mg per cow), after which L-Cit supplementation commenced at 72-h intervals. On day 10, a second PG injection (0.4 mg per cow) was administered, and supplementation was discontinued on day 11. Estrus expression was monitored, and pregnancy was assessed on day 35 post-insemination using B-mode ultrasonography. During the experimental period, six cows were randomly selected from each group. Blood samples were collected from the coccygeal vein prior to the morning feeding on days 1 and 11 using plain tubes (without anticoagulant). The samples were centrifuged at 720 × g (approximately 3,000 rpm for 15 min) to separate the serum. The obtained serum was stored at −20 °C for subsequent analysis of reproductive hormones. In addition, blood, rumen fluid, and fecal samples were obtained 10 h after mounting to support untargeted metabolomic profiling and microbial community analysis. Compared with the control group, the estrus rate in experimental Group I increased by 12.5% (p < 0.05). In experimental Group II, serum concentrations of gonadotropin-releasing hormone (GnRH) and follicle-stimulating hormone (FSH) measured 10 h after cows accepted mounting rose by 15.79% (p < 0.05) and 35.71% (p < 0.01), respectively, relative to the control group. Analysis of 16S rRNA sequences revealed significant differences in ruminal microbial taxa, including Verrucomicrobiota, Lentisphaeria, Oligosphaeraceae, vadinBE97_g_norank, vadinBE97, Eubacterium_ruminantium_group, and Prevotellaceae_g_norank. In the intestine, significant differences were observed in Lachnospiraceae, Lachnospirales, Marvinbryantia, Desulfovibrionia, Desulfovibrionaceae, and Desulfobacterota. KEGG enrichment analysis based on Liquid Chromatography-Mass Spectrometry (LC–MS) data indicated upregulation of the arginine biosynthesis pathway in the experimental groups, whereas metabolites associated with the tricarboxylic acid (TCA) cycle were notably elevated in experimental Group I. Within the experimental framework, dietary supplementation with L-Cit combined with the two-shot PG synchronization protocol reshaped the ruminal microbial community structure of Simmental cows, improved glucose metabolic efficiency and xenobiotic degradation capacity, and promoted the synthesis and release of reproductive hormones. Consequently, estrus and conception rates were enhanced.

The impact of graphene oxide (GO) nanocarbon on soil properties is mixed, with both negative and positive effects. Although it decreases the viability of some microbes, there are few studies on how its single amendment to soil or in combination with nanosized sulfur benefits soil microorganisms and nutrient transformation. Therefore, an eight-week pot experiment was carried out under controlled conditions (growth chamber with artificial light) in soil seeded with lettuce (Lactuca sativa) and amended with GO or nano-sulfur on their own or their several combinations. The following variants were tested: (I) Control, (II) GO, (III) Low nano-S + GO, (IV) High nano-S + GO, (V) Low nano-S, (VI) High nano-S. Results revealed no significant differences in soil pH, dry plant aboveground, and root biomass among all five amended variants and the control group. The greatest positive effect on soil respiration was observed when GO was used alone, and this effect remained significant even when it was combined with high nano-S. Low nano-S plus a GO dose negatively affected some of the soil respiration types: NAG_SIR, Tre_SIR, Ala_SIR, and Arg_SIR. Single GO application was found to enhance arylsulfatase activity, while the combination of high nano-S and GO not only enhanced arylsulfatase but also urease and phosphatase activity in the soil. The elemental nano-S probably counteracted the GO-mediated effect on organic carbon oxidation. We partially proved the hypothesis that GO-enhanced nano-S oxidation increases phosphatase activity.

The aim of this study was to assess the effects of organic fertilisers (cow manure + dung water; cattle slurry) applied in different annual doses of nitrogen (54 kg ha-1; 84 kg ha-1; 120 kg ha-1) according to 3 different intensities of grassland utilisation (extensive – 2 cuts per year, medium intensive – 3 cuts per year, intensive – 4 cuts per year) on percentage of plant functional groups (grasses, legumes, forbs), dry matter yield, and forage quality. The study was performed as a small-plot trial over 7 years on moderately moist grassland in the Czech Republic. The proportion of legumes was significantly higher in the fertilisation treatments with cow manure + dung-water combined with medium intensive utilisation and intensive utilisation (10.2% and 10.3%, respectively). Fertilisation significantly increased dry matter yields by 51.9 % (cow manure + dung water) and 56% (cattle slurry) compared to unfertilised controls (4.81 t ha-1). Grasslands fertilised with cattle slurry showed significantly higher concentration of crude protein (142.9 g kg-1) than unfertilised (126.4 g kg-1). Extensive grassland utilisation significantly increased crude fibre concentration (up to 282.1 g kg-1), and decreased the energy value (up to 4.68 MJ kg-1 of NEL). Organic matter digestibility was negatively influenced by extensive grassland utilisation (61.0%, 65.42 % and 67.44% for the extensive, medium intensive and intensive utilisation, respectively). Our findings suggest that medium intensive and intensive grassland utilisation using organic fertilisation which correspond to annual doses of nitrogen of 84 kg ha-1 and 120 kg ha-1, are the most suitable for animal nutrition.

The aim of this study was to evaluate the effect of mild temperature stress on ovarian activity (corpus luteum count; transferable embryos; morulae and blastocysts) in cows. The evaluation included 54 Holstein cows superovulated and flushed at one farm in the Czech Republic in three different temperature periods in flushing day (mild hot season THI = 75.66; season with optimal temperature THI = 68.85; season with lower temperature THI = 45.56). The data were analysed using the PROC GLM of Statistica®. The statistical model included the effect of season and other factors (age; condition; breeding value) on the studied reproduction traits. The results showed significantly higher ovarian activity in the cooler period of the year than in periods of mild temperature stress. There was also a non-significantly better reproduction result in younger animals under optimal condition.

Biochar application to the soil has been recommended as a carbon (C) management approach to sequester C and improve soil quality. Three-year experiments were conducted to investigate the interactive effects of three types of amendments on microbial biomass carbon, soil dehydrogenase activity and soil microbial community abundance in luvisols of arable land in the Czech Republic. Four different treatments were studied, which were, only NPK as a control, NPK + cattle manure, NPK + biochar and NPK + combination of manure with biochar. The results demonstrate that all amendments were effective in increasing the fungal and bacterial biomass, as is evident from the increased values of bacterial and fungal phospholipid fatty acid analysis. The ammonia-oxidizing bacteria population increases with the application of biochar, and it reaches its maximum value when biochar is applied in combination with manure. The overall results suggest that co-application of biochar with manure changes soil properties in favor of increased microbial biomass. It was confirmed that the application of biochar might increase or decrease soil activity, but its addition, along with manure, always promotes microbial abundance and their activity. The obtained results can be used in the planning and execution of the biochar-based soil amendments.

The aim of this study was to evaluate changes in chemical soil properties of permanent grasslands after the cessation of their regular utilisation and organic fertilisation. A long‑term small plot trial was established in 2004 in locality Rapotín. During 2004 – 2012 the experiment was fertilised with compost and slurry, both with the range of stocking rates 0.9, 1.4, and 2.0 livestock units (LU).ha−1 (corresponding to 54, 84, and 120 kg N.ha−1). It was further observed the unfertilised grassland as control. The plots were cut 2 – 4 times per year depending on given dose of fertiliser. During 2013 –2016 the regular management was ceased and the grasslands were completely abandoned. It was found statistically significant influence of the year and the type of fertiliser almost for all evaluated parameters. The dose of nitrogen was not significant. On the basis of our results we can conclude, that the both types of the organic fertilisers had a positive influence on the chemical soil properties, however, the compost manifested itself as the better fertiliser than the slurry from this point of view. In 2016, four years after the last application of the organic fertilisers, there were found the better chemical soil conditions in the treatments previously fertilised with compost.

The aim of the study was to estimate the forage quality and mycotoxin contamination in the locality of Švýcárna (1304 m a.s.l.), where the cattle grazing after the long-term management cessation was introduced on the pasture area of 3.6 ha in 2012. The pasture was divided into two grazing plots: P1 (Nar) (dominance of Nardus stricta) and P2 (Des) (dominance of Deschampsia cespitosa). Samples of forage were taken in June, July, August and September 2014 and 2015 from five permanent plots situated in different places of locality. Samples were analysed on the content of basic nutrients and macro- and micro- elements. Further, the content of deoxynivalenol (DON), zearalenone (ZEA) and T-2/HT-2 toxin was determined using ELISA method. From June to September the mean content of crude fibre significantly (P &lt; 0.05) increased (from 243.9 g.kg−1 DM to 331.7 g.kg−1 DM, respectively) and the content of crude protein significantly (P &lt; 0.05) decreased (from 166.9 g.kg−1 DM to 89.2 g.kg−1 DM, respectively). Mean concentration of DON in P1 (Nar) was numerically higher than in P2 (Des), being 715 and 620.63 µg.kg−1, respectively (P &lt; 0.05). Mean concentration of T-2/HT-2 toxin was 44.88 and 55.04 µg.kg−1 in P1 (Nar) and P2 (Des), respectively. The content of ZEA in P1 (Nar) of 54.65 µg.kg−1 was lower than in P2 (Des) being 78.72 µg.kg−1 (P &lt; 0.05).

The effects of different levels of grazing utilization (two, three and four grazing cycles per year) and mineral fertilization (nil-fertilization; N100P30K60) on the botanical composition of permanent grasslands were studied in the locality of Rapotín (Czech Republic, 332 m a.s.l.) from 2003–2010. The vegetation of the experimental pasture was classified as Cynosurion. It was found that moderate treatment (three grazing cycles per year) without mineral fertilization showed the highest value of diversity index (DI = 6.08), and maximum dominance of legumes (Dmax = 9.1%), particularly Trifolium repens. The highest dominance of grasses (Dmax = 77.7%), mainly Dactylis glomerata and Elytrigia repens, was achieved with the fertilized treatment utilized in two grazing cycles per year. Based on RDA results, tested management treatments explained 26% of species composition variability, where effect of number of grazing cycles per year was five-times higher than effect of fertilization. We recommend grassland utilization in three grazing cycles per year as the most suitable way from the objective of both species diversity and botanical composition of pastures in similar site conditions. Pasture fertilization should be more controlled by careful consideration of individual pasture goals, actual nutrient status of the soil and possible environmental risks.

This study examined the impact of cow body condition on the quality of bovine preimplantation embryos. The embryos (n = 107) were flushed from dairy cows and classified according to a five-point scale body condition score (BCS2 n = 17; BCS3 n = 31; BCS4 n = 11) on the 7th day after insemination and then analyzed for development, dead cell index (DCI), cell number and actin cytoskeleton quality. The highest embryo recovery rate (P < 0.05) was recorded in the BCS3 group and the lowest in the BCS4 group. More transferable (morula, blastocyst) embryos were obtained from the BCS4 cows (79%), compared with the BCS2 (64%) or BCS3 (63%) animals. However, cell numbers were higher in the BCS2 and BCS3 groups (P < 0.05) compared with the BCS4 embryos. Conversely, the DCI was lowest in the BCS2 (3.88%; P < 0.05) and highest in the BCS4 (6.56%) embryos. The proportion of embryos with the best actin quality (grade I) was higher in the BCS2 and BCS3 cows compared with the BCS4 group. Almost 25% of all embryos showed fragmented morphology and a higher DCI (5.65%) than normal morulas (1.76%). More fragmented embryos were revealed in the BCS2 (28.6%) and BCS4 (31.25%) groups, and less (19.15%) in the BCS3 group. The cell numbers in such embryos were lower in the BCS4 (22.57) than in the BCS2 (46.25) or BCS3 (42.4) groups. In conclusion, the body condition of dairy cows affects the quality of preimplantation embryos. A BCS over 3.0 resulted in a higher incidence of poor (fragmented) embryos.