Polo-like kinase 1 may regulate G2/M transition of mouse fertilized eggs by means of inhibiting the phosphorylation of Tyr 15 of Cdc2

Polo-like kinase 1(Plk1) has been reported to be a multifunctional kinase that plays pivotal regulatory roles in microtubule assembly during mammalian early embryonic mitosis. In the present study, we examined the expression of Plk1 at protein and mRNA level in mouse fertilized eggs by Western blot and RT-PCR. We also examined the kinase activity of Plk1. At various developmental phases of mouse one-cell stage embryos, both the protein and the mRNA of Plk1 were uniformly distributed; but the kinase activity of Plk1 increased at G2/M phase and decreased at the end of M phase. At the meantime, the phosphorylation of Tyr 15 of Cdc2 was inhibited at M phase. To investigate its function in mammalian fertilized eggs further, we used specific short hairpin RNAs (shRNA) and scytonemin, the putative inhibitor of Plk1 to suppress the activity of Plk1 in mouse fertilized eggs. Upon blockage of the activation of with Plk1 shRNA and scytonemin in mouse one-cell stage embryos, the cleavage rate decreased and the phosphorylation level of Tyr 15 of Cdc2 increased. These results imply that the Plk1 may regulate cell cycle progression of mouse fertilized eggs by means of inhibiting the phosphorylation of Tyr 15 of Cdc2.     

Follicular growth in vitro: detection of growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) during in vitro culture of ovine cortical slices

Primordial follicles from different mammal species can survive and enter the growth phase in vitro but do not develop beyond the primary stage. The hypothesis was that, in sheep, in vitro follicular growth is arrested because of a lack of secretion of GDF9 and/or BMP15. Cortical slices of 0.3-0.5 mm thickness issued from 5- to 6-month-old lambs were cultured for 15 days. The pieces were fixed on days 0, 2, 4, 7, 10, and 15 of culture. Follicle morphology, RT-PCR exploration of GDF9 and BMP15 mRNA, immunohistochemical location of their proteins and their receptor BMPRIB and BMPRII were assessed at different time of culture. The mean percentage of primordial follicles decreased from 58.6% (day 0) to 13.4% (day 15) (P<0.01), whereas that of primary follicles increased from 3.2% (day 0) to 31.5% on day 4 (P<0.01), then remained stable until day 15 (35.6%). The percentage of atretic follicles increased from 14.7% (day 0) to 27.1% (day 15) (P<0.05). A few secondary follicles were observed on days 4 and 10, representing 1.0%, and 2.1% of the total number of follicles. GDF9 and BMP15 mRNAs were detected from harvesting (day 0) up to day 15 following culture. At the same time, positive immunoreactions for GDF9, BMP15 and for BMPRIB and BMPRII were also found in oocyte cytoplasm. In conclusion, expression of GDF9, BMP15 and their receptors BMPRIB and BMPRII are detected during in vitro culture of ovine cortical slices.     

Prevalence and strain diversity of thermophilic campylobacters in cattle, sheep and swine farms

AIMS: To determine prevalence and strain diversity of thermophilic campylobacters in healthy ruminants and swine. METHODS AND RESULTS: Faecal samples collected from 343 herds (120 sheep, 124 beef cattle, 82 dairy cattle and 17 swine) in the Basque Country were screened in pools for thermophilic campylobacters. Two hundred and three herds were positive (67.1% dairy cattle, 58.9% beef cattle, 55.0% sheep and 52.9% pig), and species-specific PCR identified Campylobacter jejuni in 20.7% of the herds and Campylobacter coli in 6.4%. Campylobacter coli was isolated from the four production systems and was the most prevalent species in swine, where C. jejuni was not found. Other thermophilic campylobacters were found in all production systems. Four hundred and ninety-three animals from 11 positive herds were individually analysed, detecting significantly higher within-herd prevalences in dairy cattle (66.7%) and swine (57.8%) than in sheep (8.8%) or beef cattle (5.4%). flaA PCR-RFLP and pulsed-field gel electrophoresis analysis of a selection of isolates showed high genetic diversity. CONCLUSIONS: Healthy swine, cattle and sheep are important reservoirs of thermophilic campylobacters of different species and high genetic diversity. SIGNIFICANCE AND IMPACT OF THE STUDY: Efficient farm-based intervention measures are needed to reduce risk of infection. Non-C. jejuni/C. coli species should be monitored to investigate their significance for infection.     

Cereals level and source effects on rumen fermentation,colostrum and milk properties,and blood metabolites in periparturient ewes

Optimal type and dietary inclusion rates of cereal grains for periparturient sheep are unknown. The objective was to determine effects of feeding diets with high (H) v. low (L) levels of ground corn grain (CN) v. combined ground wheat and barley grains (WB) on intake, rumen fermentation, colostrum and milk properties, and blood metabolites of periparturient sheep. Twenty Afshari×Merino ewes were used in a completely randomized design study from 24 days prepartum through 21 days postpartum. Ewes were kept indoors in individual boxes and received once daily at 0900 h total mixed rations. Treatments were mixed rations containing either (1) H or (2) L concentrate based on either (1) 100% CN or (2) 50 : 50 ratio of ground wheat : ground barley grains in a 2×2 factorial arrangement. Each treatment group had five ewes including two twin-lamb ewes and three single-lamb ewes. Postpartal dry matter intake (DMI) increased by feeding high CN v. high and low WB, while high v. low CN improved postpartum DMI. The DMI during lambing tended to increase with the high v. low WB. Feeding CN v. WB, and feeding both CN and WB at L v. H level increased fecal pH. Postpartal rumen pH was lower with the high v. low WB (5.7 v. 6.2). Rumen concentrations of propionate were lower and of acetate were higher with L v. H grain levels. Increased dietary grain reduced urine pH for WB (7.24 v. 7.83) but not for CN (7.63 v. 7.52) prepartum. Colostrum properties, postpartal urine pH, lamb weight at birth and 21 days of age, and placental weight and expulsion time were unaffected. Milk yield increased and milk fat yield tended to increase by H v. L grain diets. Plasma glucose was increased by feeding high v. low WB, whereas CN v. WB tended to reduce peripartal plasma non-esterified fatty acids (NEFA) and increased insulin to NEFA ratio. In conclusion, more cereal grains can be included in periparturient sheep diets and CN instead of WB may be fed to periparturient sheep to improve energy status. Findings suggest opportunities to optimize periparturient ewe physiology and performance through feeding certain cereals and avoiding high levels of WB.     

Heritability of phenotypic udder traits to improve resilience to mastitis in Texel ewes

There are no estimates of the heritability of phenotypic udder traits in suckler sheep, which produce meat lambs, and whether these are associated with resilience to mastitis. Mastitis is a common disease which damages the mammary gland and reduces productivity. The aims of this study were to investigate the feasibility of collecting udder phenotypes, their heritability and their association with mastitis in suckler ewes. Udder and teat conformation, teat lesions, intramammary masses (IMM) and litter size were recorded from 10 Texel flocks in Great Britain between 2012 and 2014; 968 records were collected. Pedigree data were obtained from an online pedigree recording system. Univariate quantitative genetic parameters were estimated using animal and sire models. Linear mixed models were used to analyse continuous traits and generalised linear mixed models were used to analyse binary traits. Continuous traits had higher heritabilities than binary with teat placement and teat length heritability (h²) highest at 0.35 (SD 0.04) and 0.42 (SD 0.04), respectively. Udder width, drop and separation heritabilities were lower and varied with udder volume. The heritabilities of IMM and teat lesions (sire model) were 0.18 (SD 0.12) and 0.17 (SD 0.11), respectively. All heritabilities were sufficiently high to be in a selection programme to increase resilience to mastitis in the population of Texel sheep. Further studies are required to investigate genetic relationships between traits and to determine whether udder traits predict IMM, and the potential benefits from including traits in a selection programme to increase resilience to chronic mastitis.     

Extending the interval between second vaccination and slaughter: II. Changes in the reproductive capacity of immunocastrated ram lambs

Immunocastration improves the welfare of castrated commercial slaughter lambs; however, the time-point at which this technique influences semen quality and sperm production has not yet been established for various vaccination schedules. Furthermore, the effect of extended intervals between second vaccination and slaughter needs to be investigated regarding continued testosterone suppression in immunocastrated lambs. The effect of extending the interval between second immunocastration vaccination and slaughter from four to six weeks on the reproductive capacity of Dohne Merino lambs was examined. A total of 40 Dohne Merino lambs were stratified according to initial weight (45.4±3.68 kg) and randomly assigned to four treatments that included intact control rams (R), Burdizzo-castrated lambs (B) and lambs immunocastrated with either four (ICS4) or six (ICS6) weeks between second vaccination and slaughter. Blood and semen samples were collected throughout the study period to determine serum testosterone concentrations, evaluate semen quality and assess sperm viability. Semen samples from R showed improvement over the trial. Throughout the collection period, B lambs had low serum testosterone concentrations, poor sperm motility and sperm viability, as expected. However, a slight increase in the percentage of live sperm in semen samples from B lambs towards the end of the collection period indicated poor success rates of the technique in some lambs. Burdizzo-castration also caused testes tissue necrosis and abscessing, indicating physiological stress. Semen appearance scores varied for both immunocastrated treatments, but the mass motility scores decreased over time. The ICS6 lambs showed a consistent and continuous decline in serum testosterone concentrations and sperm viability, with an increased percentage of dead abnormal sperm in the semen samples at the end of the study. The ICS4 treatment was successful in interrupting serum testosterone production and reducing semen quality; however, not as consistently as the ICS6 treatment. Primary immunocastration vaccination influenced serum testosterone concentrations but consistently low levels were only realised for both treatments after secondary vaccination. Although all castration treatments influenced testes size and colour, the six-week vaccination-to-slaughter interval caused a greater decrease in testes cut surface L* (lightness) colour values and in seminiferous tubule circumference. Extending the interval between second immunocastration vaccination and slaughter resulted in a more consistent and reliable influence on reproductive capacity of immunocastrated lambs. Thus, immunocastration is a suitable alternative to Burdizzo-castration regarding the interruption of testosterone production and testis functioning.     

Grazing multispecies swards improves ewe and lamb performance

A two-year (2015 and 2016) grazing study was established to compare ewe and lamb performance when grazed on a perennial ryegrass only sward compared to more diverse sward types. In that study four sward types were investigated: a perennial ryegrass (Lolium perenne) only sward receiving 163 kg nitrogen per hectare per year (N/ha/yr) (PRG); a perennial ryegrass and white clover (Trifolium repens) sward receiving 90 kg N/ha/yr (PRGWC); a six species sward (two grasses (perennial ryegrass and timothy (Phleum pratense)), two legumes (white and red clover (Trifolium pratense)) and two herbs (ribwort plantain (Plantago lanceolata) and chicory (Cichorium intybus)) receiving 90 kg N/ha/yr (6S); and a nine species sward containing cocksfoot (Dactylis glomerata), greater birdsfoot trefoil (Lotus pedunculatus) and yarrow (Achillea millefolium) in addition to the six species listed above, receiving 90 kg N/ha/yr (9S). Each sward type was managed as a separate farmlet and stocked with 30 twin-rearing ewes at a stocking rate of 12.5 ewes/ha under rotational grazing management from turnout post-lambing until housing. Lamb live weight was recorded fortnightly and lambs were drafted for slaughter at 45 kg. Ewe live weight and body condition score (BCS) were recorded on five occasions annually. Lamb faecal egg count (FEC) was recorded fortnightly and lambs were treated with anthelmintics when mean lamb FEC per sward type was above 400 eggs per gram. Ewes grazing the 6S and 9S swards had heavier (P < 0.01) live weights and BCS throughout the study than the ewes grazing the PRG sward. Lambs grazing the 6S sward were heavier than lambs grazing all other sward types of 14 weeks old (P < 0.05). Lambs grazing the PRG sward required more days to reach slaughter weight than lambs grazing all other sward types (P < 0.001). Lambs grazing the 6S and 9S swards required fewer anthelmintic treatments than lambs grazing the PRG or PRGWC swards. In conclusion, grazing multispecies swards improved ewe and lamb performance and reduced the requirement for chemical anthelmintics.     

Gelsolin expression in sheep milk somatic cells during lactation

The identification of genes involved in phenotypes related to milk quality is important for both economic and health aspects in livestock production. The aim of this study was to assess the level of gelsolin gene expression in two breeds of dairy sheep – Sarda and Gentile – with pronounced differences in quantitative and qualitative milk traits. Gelsolin, a type of actin-modulating proteins is involved in the processes of actin remodeling during cell growth and apoptosis; therefore a role of this protein in mammary changes during lactation was here hypothesized. Individual milk samples were collected three times during lactation from 26 ewes of the two breeds. The differential gene expression of gelsolin in the two breeds and the three lactation times was estimated by quantitative PCR on RNA extracted from milk somatic cells. Correlations of gelsolin gene expression with milk yield and quality and days of lactation were also estimated. The results showed that gelsolin gene expression was significantly higher in the Sarda compared to the Gentile at each lactation stage, in agreement with the longer lactation duration and the higher daily milk yield of the first breed. Significant correlations of gelsolin gene expression were found with milk fat content in Sarda breed (−0.46, P<0.05). Gelsolin expression analysis confirmed the link between gelsolin gene function and milk fat content of sheep.     

A comparative study of four rumen buffering agents on productive performance,rumen fermentation and meat quality in growing lambs fed a total mixed ration

Controlling rumen fermentation using buffering agents could contribute to enhancing ruminant productivity and performance. This study was realized to investigate the effect of dietary supplementation of AcidBuf, sodium bicarbonate, calseapowder and WMC seaweed (Utva Lactuca extra) on the animal performance, volatile fatty acids, rumen pH, rumen histology and carcass characteristics of growing male Awassi lambs. A total of 60 lambs was divided into five groups. One group served as a control and fed only on a concentrate diet without any buffering, whereas the other four groups were fed the concentrate diet supplemented with 0.4% AcidBuf (Buf1), 0.4% AcidBuf plus sodium bicarbonate, 50 : 50 (Buf2), 0.4% calseapowder (Buf3) or 0.4% WMC Seaweed (Buf4) for 98 days. The feed conversion ratio was (P<0.05) improved in Buf2 compared to the control and other treatment groups. The propionic acid decreased, whereas butyric acid was increased in the treatment groups (P<0.05) compared to the control. The pH of the rumen fluid and the length of submucosa were (P<0.05) higher in Buf4 and Buf1, respectively, compared to the control. Hot and cold carcass weights were (P<0.05) higher in Buf4 compared to Buf1. Lean meat percentage and rib eye area were (P<0.05) higher in Buf4; while the fat percentage was (P<0.05) lower in Buf2 and Buf4 groups compared to the control. The lightness and yellowness of meat were (P<0.05) higher in Buf1 and Buf4 compared to the control. The meat pH was (P<0.05) higher in Buf3 and Buf4 compared to Buf2 (at 1 h) and control (at 24 h). The visceral depot fat (%) was reduced with Buf3 and Buf4 compared to the control. The results indicated that dietary supplementation of different buffering agents improved feed efficiency, rumen pH, carcass characteristics and decreased the body fat in growing Awassi lambs.