Breed and adaptive response modulate bovine peripheral blood cells’ transcriptome

Background: Adaptive response includes a variety of physiological modifications to face changes in external or internal conditions and adapt to a new situation. The acute phase proteins (APPs) are reactants synthesized against environmental stimuli like stress, infection, inflammation. Methods: To delineate the differences in molecular constituents of adaptive response to the environment we performed the whole-blood transcriptome analysis in Italian Holstein (IH) and Italian Simmental (IS) breeds. For this, 663 IH and IS cows from six commercial farms were clustered according to the blood level of APPs. Ten extreme individuals (five APP+ and APP- variants) from each farm were selected for the RNA-seq using the Illumina sequencing technology. Differentially expressed (DE) genes were analyzed using dynamic impact approach (DIA) and DAVID annotation clustering. Milk production data were statistically elaborated to assess the association of APP+ and APP- gene expression patterns with variations in milk parameters. Results: The overall de novo assembly of cDNA sequence data generated 13,665 genes expressed in bovine blood cells. Comparative genomic analysis revealed 1,152 DE genes in the comparison of all APP+ vs. all APP- variants; 531 and 217 DE genes specific for IH and IS comparison respectively. In all comparisons overexpressed genes were more represented than underexpressed ones. DAVID analysis revealed 369 DE genes across breeds, 173 and 73 DE genes in IH and IS comparison respectively. Among the most impacted pathways for both breeds were vitamin B6 metabolism, folate biosynthesis, nitrogen metabolism and linoleic acid metabolism. Conclusions: Both DIA and DAVID approaches produced a high number of significantly impacted genes and pathways with a narrow connection to adaptive response in cows with high level of blood APPs. A similar variation in gene expression and impacted pathways between APP+ and APP- variants was found between two studied breeds. Such similarity was also confirmed by annotation clustering of the DE genes. However, IH breed showed higher and more differentiated impacts compared to IS breed and such particular features in the IH adaptive response could be explained by its higher metabolic activity. Variations of milk production data were significantly associated with APP+ and APP- gene expression patterns

Serum acute phase proteins in cows with SARA (Subacute Ruminal Acidosis) suspect

The aim of this study was to evaluate the variations of Acute Phase Proteins (APPs) and other blood constituents during the onset of the sub-acute ruminal acidosis (SARA) pathological status. A total of 108 cows from 12 dairy herds were randomly selected and divided into three Groups of 36 animals each. All animals were subjected to a rumenocentesis. Group A was composed by subjects with a rumen pH>5.8, Group B was composed by subjects with a rumen pH ≤5.5≤5.8 and Group C was composed by subjects with a rumen pH<5.5. Blood samples were collected by jugular venipuncture and Haptoglobin (Hp), Serum Amyloid A (SAA), Total Proteins, Albumin and White Blood Cells (WBC) were determined. One-way ANOVA showed a statistical significance on Rumen pH, Hp, SAA. SARA seems not stimulate the APPs production from liver

In vivo assessment of the ability of condensed tannins to interfere with the digestibility of plant protein in sheep

To assess the effect of condensed tannin (CT) astringency on the digestibility of protein postruminally, CT were purified from four types (accessions) of Mulga (Acacia aneura), and a Leucaena pallida and dosed into sheep as complexes with (15)N-plant protein (tannin-protein complexes, TPC), together with an indigestible marker, chromium-EDTA (Cr-EDTA). Each CT treatment dose comprised 12 mg CT, 10 mg (15)N-protein, and 2.77 mg of Cr. A protein-only control (same as the other TPC solutions but without any CT) treatment was also included to make a total of six treatments. Treatments were applied in two 6 x 6 Latin Square designs with 72 h between each infusion for each sheep. In the first, the solutions were infused post-ruminally via an abomasal cannula. In the second they were dosed directly into the mouth following an intra-nasal dose of an analogue of vasopressin in an attempt to stimulate the oesophageal groove reflex to direct the solutions more efficiently toward the small intestine. Results showed no detectable effect of CT type on the in vivo digestibility of the (15)N-protein. Protein digestibilities were uniformly high, indicative of complete dissociation of the TPC. There was no correlation between protein digestibility, mouth to faeces, and protein digestibility, abomasum to faeces (P > 0.05). In vivo digestibility was also uncorrelated with CT astringencies defined in vitro (P > 0.05). Astringency in vitro was defined as the mg of CT required to achieve half-maximal precipitation of 0.5 mg of protein (bovine serum albumin). In vitro, the most astringent CT (A. aneura 883558), had at least 1.6 times the astringency of the weakest CT, (A. aneura 842394). The A. aneura with the weakest CT also contained less than 1/3 the total amount of CT/g leaf dry matter than that with the strongest, highlighting the scope for selection of more nutritionally useful types of A. aneura. Limitations of the in vivo protocol used are discussed and it is concluded that the effect of CT astringency on the availability of protein post-ruminally is minimal. (c) 2005 Elsevier B.V. All rights reserved

Acute phase proteins in serum and milk from dairy cows with clinical mastitis

The serum concentrations of haptoglobin, serum amyloid A and alpha1 acid glycoprotein were determined in serum collected from healthy dairy cows and cows with clinical mastitis, graded as mild (clots in milk) or moderate (clots in milk and visible signs of inflammation in the mammary gland/s) to assess their relative diagnostic value in detecting the disease. The concentrations of haptoglobin and serum amyloid A were also measured in milk collected from infected and uninfected quarters. The concentrations of haptoglobin and serum amyloid A were higher in the serum and milk from the cows with mild or moderate mastitis. The diagnostic value of haptoglobin in differentiating between healthy animals and those with mastitis gave sensitivities and specificities of 82 per cent and 94 per cent respectively with serum and 86 per cent and 100 per cent with milk. The diagnostic value of serum amyloid A in differentiating between healthy animals and those with mastitis gave sensitivities and specificities of 83 per cent and 90 per cent with serum and 93 per cent and 100 per cent with milk. The diagnostic value of serum alpha1 acid glycoprotein in differentiating between healthy animals and those with mastitis gave sensitivities and specificities of 62 per cent and 91 per cent

Effect of Manuka honey gel on the transforming growth factor β1 and β3 concentrations, bacterial counts and histomorphology of contaminated full-thickness skin wounds in equine distal limbs

ObjectiveTo investigate the effect of 66% Manuka honey gel on the concentrations of transforming growth factor (TGF)-1 and TGF-3, bacterial counts and histomorphology during healing of contaminated equine distal limb wounds.MethodsIn this experimental study of 10 Standardbred horses, five full-thickness skin wounds (2x1.5cm) were created on one metacarpus and six similar wounds were created on the contralateral metacarpus. Wounds were assigned to three groups: non-contaminated control wounds; contaminated control wounds; contaminated wounds treated daily with 1mL Manuka honey gel topically for 10 days. For the contaminated wounds, faeces were applied for 24h after wound creation. In five horses wounds were bandaged and in the other five horses wounds were left without a bandage. Biopsies were taken on days 1, 2, 7 and 10 after wounding to evaluate the effects of Manuka honey gel, wound contamination and bandaging on TGF-1 and TGF-3 concentrations, aerobic and anaerobic bacterial counts, and histomorphology.ResultsManuka honey gel had no significant effect on TGF-1 and TGF-3 concentrations or wound bacterial counts. Manuka honey gel decreased wound inflammation (days 7, 10), increased angiogenesis (days 2, 7, 10), increased fibrosis and collagen organisation (day 7) and increased epithelial hyperplasia (days 7, 10).ConclusionsTreatment with Manuka honey gel resulted in a more organised granulation tissue bed early in wound repair, which may contribute to enhanced healing of equine distal limb wounds

Grazing Soybean to Increase Voluntary Cow Traffic in a Pasture-based Automatic Milking System

Pasture-based automatic milking systems (AMS) require cow traffic to enable cows to be milked. The interval between milkings can be manipulated by strategically allocating pasture. The current experiment investigated the effect of replacing an allocation of grazed pasture with grazed soybean (Glycine max) with the hypothesis that incorporating soybean would increase voluntary cow traffic and milk production. One hundred and eighty mixed age, primiparous and multiparous Holstein-Friesian/Illawarra cows were randomly assigned to two treatment groups (n = 90/group) with a 2×2 Latin square design. Each group was either offered treatments of kikuyu grass (Pennisetum clandestinum Hoach ex Chiov.) pasture (pasture) or soybean from 0900 h to 1500 h during the experimental period which consisted of 2 periods of 3 days following 5 days of training and adaptation in each period with groups crossing over treatments after the first period. The number of cows trafficking to each treatment was similar together with milk yield (mean ≈18 L/cow/d) in this experiment. For the cows that arrived at soybean or pasture there were significant differences in their behaviour and consequently the number of cows exiting each treatment paddock. There was greater cow traffic (more cows and sooner) exiting pasture allocations. Cows that arrived at soybean stayed on the allocation for 25% more time and ate more forage (8.5 kg/cow/d/allocation) relative to pasture (4.7 kg/cow/d/allocation). Pasture cows predominantly replaced eating time with rumination. These findings suggest that replacing pasture with alternative grazeable forages provides no additional incentive to increase voluntary cow traffic to an allocation of feed in AMS. This work highlights the opportunity to increase forage intakes in AMS through the incorporation of alternative forages