Effects of continuous low dose infusion of lipopolysaccharide on inflammatory responses,milk production and milk quality in dairy cows

The objective of this study was to evaluate the effects of continuous low dose infusion of lipopolysaccharide (LPS) on inflammatory responses and milk production and quality in lactating dairy cows. Eight Holstein cows were assigned to two treatments in a cross‐over experimental design. Cows were infused intravenously either with saline solution or with saline solution containing LPS from Escherichia coli O111:B4 at a dose of 0.01 μg LPS/kg body weight for approximately 6 hr each day during a seven‐day trial. The clinical symptoms and milk production performance were observed. Milk samples were analysed for conventional components, fatty acids and amino acids. And jugular vein and mammary vein plasma samples were analysed for concentrations of cytokines and acute phase proteins. LPS infusion decreased feed intake and milk yield. An increase in body temperature was observed after LPS infusion. LPS infusion also increased plasma concentrations of interleukin‐1β, serum amyloid A, LPS‐binding protein, C‐reactive protein and haptoglobin. LPS infusion decreased the contents of some fatty acids, such as C17:1, C18:0, C18:1n9 (trans) and C18:2n6 (trans), and most amino acids except for methionine, threonine, histidine, cysteine, tyrosine and proline in the milk. The results indicated that a continued low dose infusion of LPS can induce an inflammatory response, decrease milk production and reduce milk quality.     

Effects of milk thistle meal on performance,ileal bacterial enumeration,jejunal morphology and blood lipid peroxidation in laying hens fed diets with different levels of metabolizable energy

This study was conducted to evaluate the effects of different levels of milk thistle meal on performance, blood biochemical indices, ileal bacterial counts and intestinal histology in laying hens fed diets containing different levels of metabolizable energy. A total number of 200 Leghorn laying hens (Hy‐Line W‐36) were randomly assigned to eight experimental treatments with five cage replicates of five birds each. Dietary treatments consisted of four levels of milk thistle meal (0%, 15%, 30% and 60%) and two levels of AME_n (11.09 and 12.34 MJ/kg) fed over a period of 80 days. In vitro studies revealed that the total phenolic component of milk thistle meal was 470.64 mg gallic acid equivalent/g of the sample, and its antioxidant activity for inhibiting the 2‐2‐diphenyl‐1‐picrichydrazyl free radical and reducing ferric ions was about 21% higher than that of butylated hydroxyltoluene (p < .05). Diets containing high level of AME_n led to improved egg production (p < .05), egg weight (p < .05), egg mass (p < .01) and feed conversion ratio (p < .01). In addition, offering diets containing high energy significantly enhanced (p < .01) serum triglyceride and malondialdehyde (MDA) concentrations as well as jejunal villus height. Dietary supplementation of 3% milk thistle meal resulted in the best feed conversion ratio (p < .05), reduction of ileal Escherichia coli enumeration (p < .01) and an enhancement in the villus height‐to‐crypt depth ratio (p < .05). Furthermore, feeding incremental levels of this meal led to remarkable decrease in serum cholesterol, triglyceride and MDA (p < .01) concentrations while significant increase in blood high‐density lipoprotein content and goblet cell numbers (p < .05). The present findings indicate that milk thistle meal with high antioxidant and antibacterial properties in laying hen diets may improve health indices and productive performance.     

Polyphenols in Eucalyptus leaves improved the egg and meat qualities and protected against ethanol‐induced oxidative damage in laying hens

Polyphenols in Eucalyptus leaves (PE) were value resources due to various pharmacological activities, but data on the effect on laying hens are very scare. This study was conducted to determine the effect of PE on the laying performance, egg traits, meat quality, antioxidant status and liver tissues of laying hens. One hundred and twenty 256‐day‐old Yueqinhuang laying hens were randomly assigned to four treatment groups (different levels of PE at 0, 0.5, 0.8 and 1.2 g/kg diet) for 63 days with 15 replicates per group. Addition of 0.8 g/kg PE not only improved the egg trait by increasing the eggshell thickness and decreasing MDA production and cholesterol content in the egg yolk, but also significantly enhanced meat quality of hens, as suggested by the increase pH_45 min (p < .01) and a* value, and decrease drip loss rate (p < .01). Meanwhile, under acute ethanol‐induced oxidative damage condition, supplementation of 0.8 g/kg PE enhanced the serum antioxidant status by increasing enzymatic activities (GSH‐PX, T‐SOD, T‐AOC), inhibited oxidative damage and provided protective effect to liver tissue. In conclusion, addition of 0.8 g/kg PE not only improved the egg traits and meat quality without obvious adverse effects, but also increased the serum antioxidant status of the hens and protected their liver tissue from ethanol‐induced oxidative damage. This study indicated that PE could be utilized as an effective feed additive for laying hens to improve health performance and egg traits.     

Based serum metabolomics analysis reveals simultaneous interconnecting changes during chicken embryonic development

Metabolic disorder is a major health problem and is associated with a number of metabolic diseases. Due to native hyperglycaemia and resistance to exogenous insulin, chickens as a model had used in the studies of adipose tissue biology, metabolism and obesity. But no detailed information is available about the comprehensive changes of serum metabolites at different stages of chicken embryonic development. This study employed LC/MS‐QTOF to determine the changes of major functional metabolites at incubation day 14 (E14d), 19 (E19d) and hatching day 1 (H1d), and the associated pathways of differential metabolites during chicken embryonic development were analysed using Metabolite Set Enrichment Analysis method. Results showed that 39 metabolites were significantly changed from E14d to E19d and 68 metabolites were significantly altered from E19d to H1d in chicken embryos. Protein synthesis was promoted by increasing the concentrations of L‐glutamine and threonine, and gonadal development was promoted through increasing oestrone content from E14d to E19d in chicken embryos, which indicated that serum glutamine, threonine and oestrone contents may be considered as the candidate indicators for assessment of early embryonic development. 2‐oxoglutaric acid mainly contributed to enhancing the citric cycle, and it plays an important role in improving the growth of chicken embryos at the late development; the decreasing of L‐glutamine, L‐isoleucine and L‐leucine contents from E19d to H1d in chicken embryonic development implied their possible functions as the feed additive during early posthatch period of broiler chickens to satisfy the growth. These results provided insights into understand the roles of serum metabolites at different developmental stages of chicken embryos, it also provides available information for chicken as a model to study metabolic disease or human obesity.     

Effect of feed intake level and dietary protein content on the body temperature of pigs housed under thermo neutral conditions

Feed intake and diet composition appear to affect the body temperature of pigs. Two trials were conducted to analyse the effect of feed intake level and dietary protein content on the intestinal temperature (IT) of pigs housed under thermo neutral conditions. Ten pigs (64.1 ± 1.3 kg initial body weight) fitted with an ileal cannula were used. A thermometer set to register the IT at 5‐min intervals was implanted into the ileum through the cannula. In both trials, the ambient temperature ranged from 19.1 to 21.6°C and the pigs were fed at 07:00 and 19:00 hr (same amount each time). In trial 1, the pigs were fed daily 1.2 or 1.8 kg of a wheat–soybean meal diet. The IT followed a similar pattern along a 24‐hr period regardless the feed intake level. The IT rapidly increased up to 0.61 and 0.74°C after the morning meal and up to 0.53 and 0.47°C after the evening meal in pigs fed 1.2 and 1.8 kg/d respectively. The postprandial IT was higher in pigs fed 1.8 kg after each meal (p < .05). In trial 2, pigs were fed daily 1.8 kg of a low (11%) or a high (22%) crude protein diet. The IT followed a similar pattern along the 24‐hr period regardless the dietary protein level. The postprandial IT did not differ between pigs fed the low protein or the high protein (p > .10). The IT rapidly increased up to 0.66 and 0.62°C after the morning meal in pigs fed the high‐ and low‐protein diet (p < .05), but there was no change after the evening meal (p > .10). In conclusion, the feed intake level affected the IT of pigs housed under TN conditions, but the dietary protein content had no effect.     

The effects of late gestation nutrient restriction of dams on beef heifer intake,metabolites and hormones during an ad libitum feeding trial

This study's objective was to determine if nutrient restriction during late gestation affected beef heifer feed intake, body weight (BW) gain and endocrine regulation during a 10‐week feeding trial. During the last 100 days of gestation, control (CON) dams were fed to increase body condition score (BCS). Whereas, nutrient‐restricted dams (NR) and NR dams protein supplemented 3 days/week (NRS) were fed to decrease BCS by 1.2. After parturition, all cow‐calf pairs were moved to a common pasture and fed in excess of requirements until weaning. At 15 months of age, heifers were randomly sorted into two pens and adjusted to a commercial total mixed ration over a 2‐week period. Blood samples and BW were taken at the initiation of feeding and on a biweekly basis for the duration of the feeding trial. Feed intake was monitored for 10 weeks using a GrowSafe System. After 10 weeks, an intravenous glucose tolerance test (IVGTT) was performed on 21 randomly subsampled heifers. During the feeding trial, NR heifers consumed more feed than CON and NRS heifers. Heifers from NR dams tended to increase BW compared to NRS and CON heifers when adjusted for initial BW. Heifers from NR and NRS dams had a greater increase in BCS compared to heifers from CON dams. Plasma glucose and insulin concentrations during the feeding trial increased in NR heifers compared to the other groups beginning at 2 and 4 weeks respectively. Plasma leptin concentrations were increased in the NR and NRS heifers compared to the CON heifers beginning at week 4 of feeding. During the IVGTT at the conclusion of the feeding challenge, plasma glucose and insulin were increased in NR heifers compared to other treatment groups. These results show that nutrient restriction during late gestation alters appetite and endocrine regulation in heifer offspring.     

Processing traits and digestibility of extruded dog foods with soy protein concentrate

Soya bean protein concentrate (SPC) with two particle sizes were evaluated on extrusion parameters, kibble formation, digestibility and palatability of dog foods. Eight diets were extruded: PBM—control diet based on poultry by‐product meal (PBM); GM—a diet in which corn gluten meal (GM) replaced 45% of the diet protein; cSPC15%, cSPC30% and cSPC45%—diets in which SPC of coarse particle size (600 μm) replaced 15%, 30% and 45% of the diet protein; and sSPC15%, sSPC30% and sSPC45%—diets in which SPC of small particle size (200 μm) replaced 15%, 30% and 45% of the diet protein. The digestibility of nutrients was evaluated for the PBM, GM, cSPC45% and sSPC45% diets, using six dogs per food. The PBM, GM and cSPC45% diets were compared for palatability. Data were submitted for analysis of variance, and the means were compared by polynomial contrasts or Tukey's test (p < .05). The cSPC increased the specific mechanical energy (SME) application, extrusion temperature and pressure linearly, resulting in lower kibble density and higher expansion and starch gelatinization (SG) (p < .01). When comparing the PBM, GM, cSPC45% and sSPC45% diets, higher SME, extrusion temperature and pressure, SG and kibble expansion were verified for the cSPC45% diet (p < .05). The DM, fat and crude protein digestibility were similar among diets. Faecal pH, ammonia and lactate did not differ, demonstrating that the removal of oligosaccharides and soluble non‐starch polysaccharides of SPC produces an ingredient with mostly non‐fermentable fibre. Dogs preferred the PBM to the GM diet (p < .05), but consumed the PBM and cSPC45% foods equally. In conclusion, SPC exhibited good extrusion functionality, favouring kibble expansion and SG, with high digestibility, similar to that of PBM. The removal of soluble compounds from soya beans resulted in an ingredient with low fermentable fibre content, which did not alter faecal formation or characteristics.     

Lipopolysaccharide and cytokines modulate leukotriene (LT)B4 and LTC4 production by porcine endometrial endothelial cells

Uterine inflammatory response is mediated by inflammatory mediators including eicosanoids and cytokines produced by immune and endometrial cells. Interactions between lipopolysaccharide (LPS) and cytokines, and leukotrienes (LTs) in endothelium, important for the host defence during the inflammation, are unknown. We studied the effect of LPS, tumour necrosis factor (TNF)‐α, interleukin (IL)‐1β, IL‐4 and IL‐10 on 5‐lipooxygenase (5‐LO), LTA₄ hydrolase (LTAH) and LTC₄ synthase (LTCS) mRNA and protein expression, LTB₄ and LTC₄ release from porcine endometrial endothelial cells, and cell viability. For 24 hr, cells were exposed to LPS (10 or 100 ng/ml of medium) and cytokines (each 1 or 10 ng/ml). 5‐LO mRNA/protein expression augmented after incubation with larger doses of LPS, TNF‐α, IL‐4 and IL‐10 and smaller dose of IL‐1β. Larger dose of TNF‐α, smaller doses of LPS and IL‐1β and both doses of IL‐10 increased LTAH mRNA/protein expression. LTAH protein content was up‐regulated by larger dose of LPS, but it was reduced in response to both doses of IL‐4. LTCS mRNA expression was elevated by larger doses of LPS, IL‐4 and IL‐10 or both doses of TNF‐α and IL‐1β. LTCS protein level increased after treatment with both doses of IL‐1β, IL‐4 and IL‐10, smaller dose of LPS and larger dose of TNF‐α. Both doses of LPS and larger doses of TNF‐α and IL‐10 increased LTB₄ release. LPS, IL‐1β and IL‐10 at smaller doses, or TNF‐α and IL‐4 at larger doses stimulated LTC₄ release. Smaller doses of TNF‐α and IL‐1β or both doses of IL‐4 enhanced the cell viability. This work provides new insight on the participation of LPS, TNF‐α, IL‐1β, IL‐4 and IL‐10 in LTB₄ and LTC₄ production/release from porcine endometrial endothelial cells, and the effect of above factors on these cells viability. The used cellular model gives the possibility to further establish the interactions between inflammatory mediators.     

Bovine foetal sex determination—Different DNA extraction and amplification approaches for efficient livestock production

Foetal sex determination using polymerase chain reaction (PCR) in mammals is based on the amplification of gender‐specific foetal DNA sequences circulating in maternal blood. The bovine synepitheliochorial placenta does not allow a direct contact between the trophoblast and the maternal blood, resulting in difficult passage of foetal DNA and, consequently, its very small amounts in maternal bloodstream. Circulating cell‐free foetal DNA (ccffDNA) encompasses short nucleotide fragments (300–600 bp) in maternal circulation. The aim of this study was to assess this non‐invasive method in accurate prenatal sexing in early and late gestational periods in comparison with ultrasound diagnostics. As various DNA isolation and amplification methods were tested, their success in obtaining reliable results was evaluated. Two groups were tested, each consisting of 20 pregnant cows. Blood of a bull and a non‐pregnant heifer was the controls. Extraction of foetal DNA was accomplished by three different methods: using tubes with silicone membranes, a single‐tube extraction without silicone membranes and phenol–chloroform extraction. Following each extraction method, foetal DNA was amplified using PCR and real‐time PCR with both bAML and TSPY primers in a separate reaction. Positive results were obtained only after amplification of foetal DNA extracted with a single‐tube extraction kit. In comparison with ultrasound examination results and foetal gender recorded at birth, the sensitivity of the PCR test was 90% in Group I, but the technique failed to detect male foetuses in Group II. The real‐time PCR test sensitivity in Group I was 90% and in Group II 91.6%.     

Effect of parental origin on early life history traits of European eel

Establishment of European eel (Anguilla anguilla) hatchery production will rely on selectively bred individuals that produce progeny with the best traits in successive generations. As such, this study used a quantitative genetic breeding design, between four females and nine males (four wild‐caught and five cultured), to investigate the effect of paternal origin (wild‐caught vs. cultured) and quantify the relative importance of parental effects, including genetic compatibility, on early life history (ELH) performance traits (i.e. fertilization success, embryonic survival at 32 hr post‐fertilization, hatch success and larval deformities at 2 days post‐hatch) of European eel. Wild‐caught males had higher (56%) spermatocrit values than cultured males (45%), while fertilization success, embryonic survival, hatch success and larval deformities were not significantly impacted by paternal origin. This demonstrates that short‐term domestication of male eels does not negatively affect offspring quality and enables the consideration of cultured male broodstock in future breeding programmes. Moreover, paternity significantly explained 9.5% of the variability in embryonic survival, providing further evidence that paternal effects need to be taken into consideration in assisted reproduction protocols. Furthermore, maternity significantly explained 54.8% of the variation for fertilization success, 61.7% for embryonic survival, 88.1% for hatching success and 62.8% for larval deformities, validating that maternity is a major factor influencing these “critical” ELH traits. At last, the parental interaction explained 12.8% of the variation for fertilization success, 8.3% for embryonic survival, 4.5% for hatch success and 20.5% for larval deformities. Thus, we conclude that eggs of one female can develop more successfully when crossed with a compatible male, highlighting the importance of mate choice for successful propagation of high‐quality offspring. Together, this knowledge will improve early offspring performance, leading to future breeding programmes for this critically endangered and economically important species.