Label‐free proteome of water buffalo (Bubalus bubalis) seminal plasma

The study aimed to describe the Bubalus bubalis seminal plasma proteome using a label‐free shotgun UDMS^E approach. A total of 859 nonredundant proteins were identified across five biological replicates with stringent identification. Proteins specifically related to sperm maturation and protection, capacitation, fertilization and metabolic activity were detected in the buffalo seminal fluid. In conclusion, we provide a comprehensive proteomic profile of buffalo seminal plasma, which establishes a foundation for further studies designed to understand regulation of sperm function and discovery of novel biomarkers for fertility. MS data are available in the ProteomeXchange with identifier PXD003728.     

Jellyfish as an Alternative Source of Bioactive Antiproliferative Compounds

Jellyfish are commonly considered a nuisance for their negative effects on human activities (e.g., fisheries, power plants and tourism) and human health. However, jellyfish provide several benefits to humans and are commonly eaten in eastern countries. Additionally, recent studies have suggested that jellyfish may become a source of high-value molecules. In this study, we tested the effects of the methanolic extracts and enriched fractions, obtained by solid-phase extraction fractionation, from the scyphomedusae Pelagia noctiluca, Rhizostoma pulmo, Cotylorhiza tuberculata and the cubomedusa Caryddea marsupialis on different human cancer cell lines in order to evaluate a potential antiproliferative activity. Our results indicated that fraction C from Caryddea marsupialis-(CM) and C. tuberculata oral arms (CTOA) were the most active to reduce cell viability in a dose-dependent manner. LC/MS based dereplication analyses highlighted that both bioactive fractions contained mainly fatty acids and derivatives, with CM additionally containing small peptides (0.7–0.8 kDa), which might contribute to its higher biological activity. The mechanism of action behind the most active fraction was investigated using PCR arrays. Results showed that the fraction C of CM can reduce the expression of genes involved in apoptosis inhibition in melanoma-treated cells, which makes jellyfish a potential new source of antiproliferative drugs to be exploited in the future.     

Effect of restricted suckling on milk yield,composition and flow,udder health,and postpartum anoestrus in grazing Holstein cows

The effect of restricted suckling on milk yield and composition, udder health, and postpartum anoestrus in dairy cows in pasture-based systems, was studied in 32 Holstein multiparous cows and their calves. At calving, each cow–calf pair was randomly assigned to one of two treatments: restricted suckling (RS) of the cows by her own or another calf, twice daily for 30 min or artificial rearing (AR) of the calves with milk obtained from the bulk tank, offered twice a day in buckets. Treatments were applied until week 8 after calving. The diet of the cows consisted of direct grazing in improved pastures, corn silage and a commercial concentrate which was offered at milking. Milk production and composition, udder health, body condition score of the cows, body weight and milk intake of the calves were measured weekly, and the first postpartum ovulation was determined three times a week by ovarian ultrasonography. Cows with RS management had a lower machine-milked milk yield (17.9 vs. 24.8 kg/d), a lower fat percentage (3.21 vs. 4.11%) and 4% fat-corrected milk yield (16.2 vs. 25.7 kg/d), and also a lower average milk flow (1.35 vs. 1.76 kg/min) than cows in the AR treatment. There was no effect of treatment on milk protein percentage or udder health as measured by milk electrical conductivity. The interval from calving to first postpartum ovulation was shorter in the AR cows than in the RS cows (18.5 vs. 21.8 days). The RS calves consumed more milk (7.2 vs. 5.4 kg/d), gained more body weight (0.813 vs. 0.656 kg/d), and had a higher body weight at weaning (84.3 vs. 73.3 kg) than AR calves. Restricted suckling of grazing dairy cows had a negative effect on machine-milked milk yield, fat percentage and 4% fat-corrected milk yield, but had no effect on udder health or on improved weight gain and body weight at weaning of the calves.     

The blastocyst production rate and incidence of chromosomal abnormalities by developmental stage in in vitro produced porcine embryos

The present study was conducted to determine the relationship between embryonic development speed at different stages (the cleaved stage at 52 h and the blastocyst stage at 6 days post insemination) and incidences of chromosome abnormalities in in vitro produced porcine embryos. Porcine oocytes were collected from 3-6-mm ovarian follicles obtained at a slaughterhouse and matured in modified NCSU-37 medium for 44-46 h. Following in vitro fertilization with a final concentration of 1 x 10(5) sperm/ml for 3 h, all oocytes were cultured in vitro for 52 h. Day-2 (52 h after insemination) embryos were classified according to their cleaved stages into 2-cell, 3- to 4-cell, 5- to 8-cell, and >8-cell stages; these were cultured separately for additional 4 days (Day 6). The resultant Day-6 blastocysts were classified according to the morphological diameter into 3 grades: Grade A, expanded blastocysts; Grade B, expanding blastocysts; and Grade C, early blastocysts. They were then analyzed chromosomally. The 3- to 4-cell and 5- to 8-cell embryos had significantly high blastocyst development rates (46.1 and 36.9%, respectively), and these blastocysts contained significantly more cells (40.2 and 42.4 cells, respectively) than those derived from 2-cell embryos and >8-cell embryos (28.6 and 26.5 cells, respectively). The incidence of chromosomal abnormalities was significantly higher in the blastocysts derived from 2-cell and >8-cell stage embryos than in the blastocysts derived from the other stage embryos. Furthermore, the grade A blastocysts had the lowest incidence of chromosomal abnormalities (35.3%) and contained the most cells (48.7 cells). Porcine in vitro production (IVP) yielded a high blastocyst rate and an excellent embryo quality when 3- to 4-cell and 5- to 8-cell stage embryos were selected on Day 2 after insemination. The same criteria yielded a higher quality of expanded blastocysts based on the stage of embryo development and morphology.     

Seroepidemiology of beef and dairy herds and fetal study of Neospora caninum in Argentina

The purpose of the present work was to study the epidemiology of Neospora caninum in beef and dairy herds in the Humid Pampas of Argentina. The seroprevalence of N. caninum was evaluated in 2414 serum samples of cows from beef and dairy farms. An indirect fluorescent antibody test (IFAT) was used to determine specific antibodies. The sera was screened at a dilution >or=1:200 and >or=1:600 in cows with reproductive disease antecedents and without them, respectively. Cows without history of reproductive diseases from nine beef and fifteen dairy farms were grouped according to the percentage (> or or 相似文献   

Effects of different variables on whole blood cholinesterase analysis in dogs.

The influence of several variables such as sample and reagent storage, anticoagulants, reaction temperature, pH, and substrate concentration on whole blood cholinesterase determination was studied. Storage of nondiluted whole blood samples at room temperature or under refrigeration (4 C) was adequate for short-term storage (3 days to 2 weeks). However, freezing would be more appropriate for long-term storage (> or = 1 month), and successive thawing and freezing did not produce any loss of cholinesterase activity. All reagents (2,2'-dithiodipyridine as chromophore and acetylthiocholine and butyrylthiocholine as substrates) were stable for 3 months when frozen. Heparin and ethylenediaminetetraacetic acid were the most suitable anticoagulants for whole blood acetylcholinesterase and butyrylcholinesterase determination, because citrate yielded lower acetylcholinesterase values and fluoride inhibited butyrylcholinesterase. Increases in reaction temperature and pH yielded higher cholinesterase values but also increased nonenzymatic substrate hydrolysis. Higher cholinesterase and nonenzymatic substrate hydrolysis values were obtained as higher substrate concentrations were used.     

Porcine monocyte subsets differ in the expression of CCR2 and in their responsiveness to CCL2

Monocyte subsets have been shown to differ in the pattern of chemokine receptor expression and their migratory properties, both in human and mouse. Previously we have characterized in the swine several monocyte subpopulations, based on the expression of CD163, Tük4 and SLA-II, which share features with the populations described in human and mouse. Here, we have analysed the expression of different chemokine receptors in the CD163⁻Tük4⁺SLA-II⁻ and CD163⁺Tük4⁻SLA-II⁺ populations of porcine monocytes. CD163⁺Tük4⁻SLA-II⁺ monocytes expressed higher CX₃CR1 but lower CCR2 and CXCR4 mRNA levels than CD163⁻Tük4⁺SLA-II⁻ monocytes. Moreover, porcine CCL2 binding on Tük4⁺SLA-II⁻ but not on Tük4⁻SLA-II⁺ cells was detected by using a CCL2-green fluorescence protein (pCCL2-GFP) fusion protein. Finally, flow cytometric analyses of monocytes recovered after chemotaxis assays show a clear increase in the proportion of Tük4⁺SLA-II⁻ cells in the fraction migrating toward CCL2, consistent with the polarized CCR2 expression in this monocyte population. The pattern of expression of these chemokine receptors reinforces the similarities of these porcine subsets with their human and mouse counterparts.     

Use of quantitative two-dimensional color tissue Doppler imaging for assessment of left ventricular radial and longitudinal myocardial velocities in dogs

OBJECTIVE: To determine left ventricular free wall (LVFW) radial and longitudinal myocardial contraction velocities in healthy dogs via quantitative 2-dimensional color tissue Doppler imaging (TDI). ANIMALS: 100 dogs. PROCEDURE: TDI was used by a single trained observer to measure radial and longitudinal myocardial movement in the LVFW. Radial myocardial velocities were recorded in segments in the endocardial and epicardial layers of the LVFW, and longitudinal velocities were recorded in segments at 3 levels (basal, middle, apical) of the LVFW. RESULTS: LVFW velocities were higher in the endocardial layers than in the epicardial layers. Left ventricular free wall velocities were higher in the basal segments than in the middle and apical segments. Radial myocardial velocity gradients, defined as the difference between endocardial and epicardial velocities, were (mean +/- SD) 2.5 +/- 0.8 cm/s, 3.8 +/- 1.5 cm/s, and 2.3 +/- 0.9 cm/s in systole, early diastole, and late diastole, respectively. Longitudinal myocardial velocity gradients, defined as the difference between basal and apical velocities, were 5.9 +/- 2.2 cm/s, 6.9 +/- 2.5 cm/s, and 4.9 +/- 1.7 cm/s in systole, early diastole, and late diastole, respectively. A breed effect was detected for several systolic and diastolic TDI variables. In all segments, systolic velocities were independent of fractional shortening. CONCLUSIONS AND CLINICAL RELEVANCE: LVFW myocardial velocities decreased from the endocardium to the epicardium and from base to apex, thus revealing intramyocardial radial and longitudinal velocity gradients. These indices could enhance conventional echocardiographic analysis of left ventricular function in dogs. Breed-specific reference intervals should be defined.     

Identification of common QTL for resistance to <Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis> in three doubled haploid populations of <Emphasis Type="Italic">Brassica napus</Emphasis> (L.)

Sclerotinia stem rot (SR) is one of the most devastating diseases of canola/rapeseed. Quantitative trait loci (QTL) analyses were carried out to identify loci responsible for resistance to SR in three doubled haploid DH populations (H1, H2 and H3). Petiole inoculation technique PIT was used to evaluate the all populations for resistance to SR. Genetic maps were developed using sequence related amplified polymorphism SRAP and simple sequence repeat SSR markers. Genetic maps of the H1 and H2 populations were developed using 508 and 478 markers, respectively. Previously published genetic map of the H3 population was also used in this study. The QTL analysis was carried out for each replicate separately as well as on the average of all the replicates. The numbers of identified QTL in each analysis varied from four to six in the H1 population, three to six in the H2 population and two to six in the H3 population. A number of common QTL were identified between the replicates of each population. Two common QTL were identified on linkage group A7 and C6 between the H1 and H3 populations and one QTL on A9 between the H2 and H3 populations. We are the first to report, identification of common QTL between different populations of Brassica napus.