Protein Composition of Seminal Plasma in Fractionated Stallion Ejaculates

Seminal plasma (SP) contains several types of compounds derived from the epididymides and accessory glands. The aim of this study was to examine the protein composition of different ejaculate fractions. Trial I: fractionated ejaculates were collected from two normal and two subfertile stallions. Samples containing pre‐sperm fluid and the first sperm‐rich jets (HIGH‐1), the main sperm‐rich portion (HIGH‐2), the jets with low sperm concentrations (LOW), and a combined whole‐ejaculate (WE) sample was centrifuged, and the SP was filtered and frozen. A part of each SP sample was stored (5°C, 24 h) with spermatozoa from HIGH‐2 and skim milk extender. Sperm motility was evaluated after storage in extender mixed with the stallion’s own SP or SP from one of the other stallions (sperm from a normal stallion stored in SP from a subfertile stallion and vice versa). Protein composition was analysed using reverse‐phase liquid chromatography (RP‐HPLC), N‐terminal sequencing and mass spectrometry. The area‐under‐the‐curve (AUC) was used for quantitative comparison of proteins within fractions. Trial II: semen samples were collected from seven stallions. Fractions with the highest (HIGH) and lowest (LOW) sperm concentrations and WE samples were examined using SDS‐PAGE and densitometry. No significant differences emerged between fractions in the AUC‐values of the Horse Seminal Protein‐1 (HSP‐1) and HSP‐2 peaks, or the peak containing HSP‐3 and HSP‐4 (HSP‐3/4). Levels of HSP‐1, HSP‐2 and HSP‐3/4 were not significantly correlated with total sperm motility, progressive sperm motility or average path velocity after storage. Significant differences between ejaculate fractions in the amount of different protein groups present in SP were not found in Trial I; but in Trial II, the proteins in the 60–70 kDa range were more abundant in LOW than in HIGH and WE, indicating that this band contained proteins derived mainly from the seminal vesicles, which produce most of the SP in LOW.     

Postprandial glycemia, insulinemia, and satiety responses in healthy subjects after whole grain rye bread made from different rye varieties. 1

Rye products typically induce low insulin responses and appear to facilitate glucose regulation. The objective of this study was to investigate differences in postprandial glucose, insulin, and satiety responses between breads made from five rye varieties. Breads made from whole grain rye (Amilo, Rekrut, Dankowski Zlote, Nikita, and Haute Loire Pop) or a white wheat bread (WWB) were tested in a randomized cross-over design in 14 healthy subjects (50 g available starch). Metabolic responses were also related to the composition of dietary fiber and bioactive compounds in the breads and to the rate of in vitro starch hydrolysis. The Amilo and Rekrut rye breads induced significantly lower insulin indices (II) than WWB. Low early postprandial glucose and insulin responses (tAUC 0-60 min) were related to higher amounts of caffeic, ferulic, sinapic, and vanillic acids in the rye breads, indicating that the phenolic acids in rye may influence glycemic regulation. All rye breads induced significantly higher subjective feelings of fullness compared to WWB. A low II was related to a higher feeling of fullness and a lower desire to eat in the late postprandial phase (180 min). The data indicate that some rye varieties may be more insulin-saving than others, possibly due to differences in dietary fiber, rate of starch hydrolysis, and bioactive components such as phenolic acids.     

Intestinal gene expression in pigs experimentally co-infected with PCV2 and PPV

The objective of the present study was to characterize the local immune reaction in the intestine of pigs experimentally infected with PCV2 and PPV. Archived intestinal material from an experimental study in which pigs were co-infected with a Swedish isolate of PCV2 (S-PCV2) and PPV, or a reference isolate of PCV2 (PCV2-1010) and PPV, were used. The intestinal samples were analysed by qPCR for expression of a number of selected cytokines and the overall gene expression in the intestine was screened by cDNA microarray. Analyses by qPCR showed that pigs infected with PCV2-1010/PPV displayed a significantly increased mRNA expression for IL-6 (p<0.05), IL-10 (p<0.05) and IFN-γ (p<0.05). The microarray screening revealed a strong up-regulation of IFITM3 along with several other interferon-stimulated genes (ISGs) in pigs infected with PCV2/PPV. The analyses also indicated differences between the two isolates. Fewer pigs infected with S-PCV2/PPV expressed the cytokines detected by qPCR, compared to pigs infected with PCV2-1010/PPV, and pigs infected with S-PCV2/PPV displayed a higher proportion of down-regulated genes than PCV2-1010/PPV-infected pigs.     

Measurement and fissuring of rice kernels during quasi-moisture sorption by image analysis

Fissuring associated with kernel elongation was evident in a moisture-adsorbing environment at 15 °C of relative humidity 88.72±0.28% with moisture content from 6% (d.b.) to 16.3% (d.b.). The average length and width of 800 kernels was measured during the sorption test using a Cervitec Grain Inspector equipped with two digital cameras. Samples were exposed to both a moisture-adsorbing and desorbing environments at 15 °C with relative humidities of 23.4±0.53%, 55.9±0.27%, 70.98±0.28% and 88.72±0.28%. On image analysis, the kernels changed in size from 5.13 to 4.84 mm in length and 2.9 to 2.73 mm in width. Both length and width changed as expected during moisture-adsorbing and moisture-desorbing tests. Fissured kernel percent was related to changes in the average length of the kernels during the moisture-adsorbing tests. The average length and fissured kernel percent of rice kernels measured by image analysis may be considered as a reliable indicator of changes in their dimensions during quasi-static moisture sorption processes.     

Mixotrophy in a Local Strain of Nannochloropsis granulata for Renewable High-Value Biomass Production on the West Coast of Sweden

A local strain of Nannochloropsis granulata (Ng) has been reported as the most productive microalgal strain in terms of both biomass yield and lipid content when cultivated in photobioreactors that simulate the light and temperature conditions during the summer on the west coast of Sweden. To further increase the biomass and the biotechnological potential of this strain in these conditions, mixotrophic growth (i.e., the simultaneous use of photosynthesis and respiration) with glycerol as an external carbon source was investigated in this study and compared with phototrophic growth that made use of air enriched with 1–2% CO₂. The addition of either glycerol or CO₂-enriched air stimulated the growth of Ng and theproduction of high-value long-chain polyunsaturated fatty acids (EPA) as well as the carotenoid canthaxanthin. Bioassays in human prostate cell lines indicated the highest antitumoral activity for Ng extracts and fractions from mixotrophic conditions. Metabolomics detected betaine lipids specifically in the bioactive fractions, suggesting their involvement in the observed antitumoral effect. Genes related to autophagy were found to be upregulated by the most bioactive fraction, suggesting a possible therapeutic target against prostate cancer progression. Taken together, our results suggest that the local Ng strain can be cultivated mixotrophically in summer conditions on the west coast of Sweden for the production of high-value biomass containing antiproliferative compounds, carotenoids, and EPA.     

Effects of drinking water temperatures on water intake and milk yield of tied-up dairy cows

The effect of four drinking water temperatures (3, 10, 17 and 24°C) on water intake, feed consumption, milk yield and composition, live weight, salt consumption, and rumination, was studied with eight tied-up dairy cows of the Swedish Red and White Breed. The cows were in their 12th to 15th week of lactation at the start of the experiment. A Latin Square design was used with four periods of three weeks each in length. The cows were fed 9 kg hay, 2 kg dried molassed beet pulp and 9 kg concentrates throughout the experiment. The study was conducted from March until June in an average daily barn temperature of 15.3°C (10.2–23.7°C).Water intake was 75.6, 76.7, 76.9 and 71.5 1/day for 3, 10, 17 and 24°C respectively. The intake of the warmest water differed significantly (P<0.01) from the others. The corresponding figures for the milk yield were 25.39, 25.93, 26.33 and 26.09 kg 4% fat-corrected milk per day, where a t-test showed a significance of P<0.05 between 3 and 10°C, P<0.001 between 3 and 17°C and P<0.01 between 3 and 24°C. Feed and salt consumption, milk composition, live weight and rumination were not affected by treatment.     

Prenatal diagnosis of bovine citrullinaemia

Holstein/Friesian embryos, progeny of citrullinaemia heterozygotes, were transferred to recipient cows. The citrullinaemia genotype of the fetus/calf was established, at birth, by estimation of citrulline concentration in plasma or by analyses of argininosuccinate synthetase exon 5 sequences in dna from blood leucocytes. Between 115 and 125 days of gestation amniotic fluid was collected transabdominally from nine recipient cows. The mean citrulline concentration in amniotic fluid from fetuses unaffected with citrullinaemia was 28 μM, SD 9 and range 15 to 38 μM. Citrulline concentration was 107 and 130 μM in amniotic fluid from two fetuses homozygous for the citrullinaemia mutation. Only the normal bovine citrullinaemia sequence was detected in cells cultured from amniotic fluid taken from two homozygous normal and two heterozygous fetuses. Both normal and mutant sequences were found in cells cultured from amniotic fluid collected from three of five heterozygous fetuses. Only the mutant sequence was found in cells cultured from the two citrullinaemia affected fetuses. mhc class II DRB3 haplotyping revealed that in four instances cultured cells were from the surrogate mother, not the fetus, these included samples from two heterozygotes in which only the normal sequence was detected in amniotic fluid.