Expression of tumor necrosis factor-alpha and cyclooxygenase-2 mRNA in porcine split-thickness wounds treated with epidermal growth factor by quantitative real-time PCR

Epidermal growth factor (EGF) accelerates the re-epithelialization of damaged epidermal cell layers in a wound, so it especially shortens the duration of wound healing. The effect of EGF on pro-inflammatory cytokines, tumor necrosis factor-alpha (TNF-alpha) and cyclooxygenase-2 (COX-2), levels during wound healing has not been reported. We investigated the relationship between exogenous EGF treatment and the expression of TNF-alpha and COX-2 mRNA in porcine split-thickness wounds by real-time PCR. Twenty split-thickness wounds were created on the back of five pigs. Fifteen wounds were treated twice daily with EGF ointments (1 microg/g, 10 microg/g, and 50 microg/g) for 10 days and five wounds were untreated. Healing time until full-epithelialization was evaluated. We performed a quantitative analysis of TNF-alpha and COX-2 mRNA expression in wound biopsies using real-time PCR. Topical application of 1 microg/g EGF accelerated re-epithelialization more than treatments of EGF at 10 microg/g and 50 microg/g, and no treatment. The levels of TNF-alpha and COX-2 mRNA were significantly greater in wounds treated with 1 microg/g than those with 10 microg/g and 50 microg/g EGF, and no treatment. Topical treatment of EGF influences the level of TNF-alpha and COX-2 mRNA within porcine split-thickness wounds. EGF-dependent slightly up-regulation of TNF-alpha and COX-2 mRNA expression during the inflammatory phase of healing may create an optimal molecular environment for wound healing.     

Effect of dietary boron on physiological responses in growing steers inoculated with bovine herpesvirus type-1

Thirty-six Angus and Angus × Simmental steers were fed one of three dietary treatments; (1) control (no supplemental B), (2) 5 mg supplemental B/kg, and (3) 15 mg supplemental B/kg for 47 days to determine the effects of dietary boron (B) on disease resistance following an inoculation with bovine herpesvirus type-1 (BHV-1). On day 34 of the study steers were inoculated intranasally with BHV-1. Rectal temperatures began to elevate at day 2, and plasma tumor necrosis factor-α concentrations increased (P < 0.05) by day 2 following BHV-1 inoculation. Plasma acute phase proteins were increased (P < 0.01) while plasma interferon-γ was decreased (P < 0.05) by day 4 post-inoculation. Supplementation of B increased (P < 0.001) plasma B concentrations in a dose-responsive manner. However, dietary B did not affect the duration and severity of clinical signs of BHV-1 and had minimal effects on plasma acute phase proteins and cytokines.     

Relationship between serum acute phase protein concentrations and lesions in finishing pigs

To establish the relationship between serum levels of three acute phase proteins, haptoglobin (Hp), C-reactive protein (CRP) and serum amyloid A (SAA), and the occurrence and severity of lesions at slaughter, a study was carried out using 70 fattening pigs from a finishing unit. Pigs were divided into three groups: Group 1 (25 pigs with clinical signs of disease), Group 2 (25 apparently healthy pigs with lesions at slaughter) and Group 3 (20 apparently healthy pigs with no lesions at slaughter). Serum levels of CRP, SAA and Hp were significantly higher in pigs with clinical signs of disease than in apparently healthy animals. Additionally, in apparently healthy pigs, serum levels of Hp and CRP were significantly higher in animals with lesions than those without lesions. The extent and severity of lung lesions were related to serum levels of Hp.     

Effects of sub-clinical post-weaning colibacillosis and dietary protein on acute phase proteins in weaner pigs

There is an increasing body of evidence that plasma acute phase proteins (APP) can be used as a marker to assess pig health. The objective of this study was to assess whether experimentally induced sub-clinical post-weaning colibacillosis (PWC) affects the concentration of two plasma APP, i.e. haptoglobin (Hp) and C-reactive protein (CRP). Since sub-clinical PWC is sensitive to protein supply, we additionally assessed whether these APP responses are sensitive to dietary protein contents. Weaned pigs (6 groups of 9 each) were offered one of the three foods, i.e. H (230 g CP/kg), L (130 g CP/kg) or HA (23% CP with anti-microbial growth promoters). On day 5 post-weaning, pigs were either dosed with 10⁸ colony forming units of enterotoxigenic E. coli or were sham-infected. Three days later and across feeding treatments, infection had reduced feed intake. Feeding and infection treatment significantly interacted for both Hp and CRP. Infection increased Hp concentration in H and HA pigs, but not in L pigs, whilst infection increased CRP concentration to a larger extent in HA pigs than in H and L pigs. The results suggest that sub-clinical PWC leads to changes in APP profiles and that these changes may be sensitive to dietary protein contents. Usefulness of APP to describe health status in pigs may therefore need to be considered in relation to their nutritional environment.     

Immunoenhancing effect of trans-10, cis-12 conjugated linoleic acid on the phagocytic capacity and oxidative burst activity of canine peripheral blood phagocytes

The effect of trans-10, cis-12 conjugated linoleic acid (t10c12-CLA) on the phagocytic capacity and oxidative burst activity (OBA) of canine peripheral blood phagocytes was examined. t10c12-CLA did not directly affect the phagocytic capacity and OBA of peripheral blood mononuclear cells (PBMC), monocytes or polymorphonuclear cells (PMN). However, the phagocytic capacity of PMN and monocytes was enhanced by the culture supernatant from t10c12-CLA-treated PBMC. This supernatant enhanced the latex bead-induced OBA of PMN and monocytes. t10c12-CLA also increased TNF-alpha production by PBMC. Recombinant canine (rc) TNF-alpha also increased the phagocytic capacity and OBA of PMN and monocytes. The ability of the culture supernatant from t10c12-CLA-treated PBMC to stimulate the phagocytic capacity and OBA of phagocytes was inhibited by anti-rcTNF-alpha pAb. These results suggest that t10c12-CLA has an immunoenhancing effect on the phagocytic capacity and OBA of phagocytes, and this effect may be mediated by TNF-alpha released from t10c12-CLA-treated PBMC.     

Potential use of hematological and acute phase protein parameters in the diagnosis of acute Schmallenberg virus infection in experimentally infected calves

The initial viraemic phase of Schmallenberg virus (SBV) infection in bovine animals is characterized by the non-specific and inconspicuous clinical signs of pyrexia (>40 °C), drop in milk yield and sometimes diarrhea. As a result, the early detection of SBV epizootics can difficult, and typically only become apparent when the congenital form of the disease is observed. The aim of the study was to describe the course of the acute phase response and haematological findings in bovine calves following experimental SBV infection. No clinical signs except for increase in rectal temperature were observed in the calves inoculated subcutaneously with a Polish strain of SBV. Viral RNA was detected in serum at 2 and 4 days post inoculation (dpi). SBV antibodies were first detected by ELISA (9–21 dpi), and subsequently by virus neutralization test (14–32 dpi). The hematological parameters showed a reduction in mid-size leucocytes (MID), and also in red blood cell count (RBC). An increase in mean corpuscular hemoglobin was also observed in SBV infected calves. No significant difference in acute phase proteins (APP) was observed between experimentally infected and control calves, suggesting limited potential as diagnostic biomarker of acute SBV infection.     

Correlations between carcass traits and mRNA levels of CGI-105 and CCAAT/enhancer protein α genes in steers of Korean cattle

To understand the molecular mechanisms that regulate intramuscular fat deposition (marbling), cDNA clones expressed in adipose tissues of Korean cattle were identified and characterized. One clone had a total length of 1262 nucleotides coding for 314 amino acids. It was identified as one encoding bovine homolog of human CGI-105 mRNA. CGI-105 is a member of fumarylacetoacetate hydrolase family. Comparison of the deduced amino acid sequences of bovine CGI-105 with those of human revealed more than 89% identity. High levels of CGI-105 mRNA expression were detected in muscle, heart, and kidney tissues among various bovine tissues. Carcass traits, including backfat thickness, rib eye area, yield index, marbling score, and quality grade were analyzed in steer of Korean cattle. A CCAAT/enhancer binding protein alpha (C/EBPα) is one of adipocyte differentiation factors that may affect deposition of fat in muscle. mRNA levels of CGI-105 and C/EBPα genes were determined in the loin muscle tissues of steers. Correlation between carcass traits and mRNA levels of the genes was estimated by Pearson's correlation coefficient. The mRNA levels of C/EBPα showed strong positive correlation (r = 0.83, p < 0.01) with marbling scores. The results of the present study indicate that the manipulation of the expression of the C/EBPα gene may contribute to the development of a method for enhancing intramuscular fat deposition in beef.     

Serum α‐1‐acid glycoprotein concentration in clinically healthy puppies and adult dogs and in dogs with various diseases

Background: α‐1‐acid glycoprotein (AGP) is an acute‐phase protein and a serum marker of inflammation and neoplasia in humans. AGP concentrations in diseased dogs and the potential effects of age, breed, and sex have not been elucidated. Objective: The purpose of this study was to examine differences in AGP concentration based on age, sex, and breed in a large population of clinically healthy dogs and to compare AGP concentrations in dogs with various diseases. Methods: Serum was obtained from clinically healthy puppies (n=74) and adults (n=172) of both sexes, and included mongrels (n=205) and Beagles (n=41). Serum also was obtained from 192 dogs with various diseases, including 8 with pyometra that were sampled before, and 1, 2, 3, and 10 days after surgery. AGP concentration was measured by single radial immunodiffusion. Statistical comparisons were made among age, sex, breed, and disease groups. Results: Serum AGP in healthy adult mongrels was 364±106 mg/L (reference interval, 152–576 mg/L). AGP was lowest in newborns (n=11, 122±54 mg/L) and gradually increased to adult levels by 3 months of age. Median AGP concentration was highest in dogs with parvovirus (n=17, 2100 mg/L), distemper (n=7, 1250 mg/L), and pyometra (n=18, 2480 mg/L) and was also significantly higher in dogs with acute filariasis, renal failure, urolithiasis, pancreatitis, hepatitis, trauma, hyperadrenocorticism, and immune‐mediated hemolytic anemia. Dogs with acute filariasis and acute hepatopathy had significantly higher AGP concentrations than dogs with chronic filariasis and chronic hepatopathy. Serum AGP concentration decreased gradually following surgery for pyometra but remained increased after 10 days (896±175 mg/L). Conclusions: Because of significantly lower AGP in puppies, the age of dogs should be considered when using AGP as a marker of disease. Serum AGP may be a useful marker of inflammatory disease in dogs and may help differentiate acute and chronic stages of disease.     

Variation in the expression of Hsp27, αB-crystallin mRNA and protein in heart and liver of pigs exposed to different transport times

Twenty pigs were randomly divided into four groups of five pigs each (not transported – control, 1, 2 and 4 h of transportation). A significant increase of ALT, AST and CK in the blood serum and acute parenchyma cell lesions were observed and those were characterized by acute degenerations in the heart and liver. Hsp27 expression levels increased significantly in the heart after 2 h and in the liver after 4 h of transportation, accompanying with the hsp27 mRNA increasing significantly in the heart and liver after 1 h of transportation. αB-crystallin expression levels were fluctuant (not significantly) in the heart and liver during transporting, however, αB-crystallin mRNA increase notably in the heart after 1 h and decrease significantly in the liver at 1 and 2 h of transportation, respectively. In conclusion, the cellular damage to the heart and liver is highest after 1 h of transportation, Hsp27 and αB-crystallin play dissimilar roles and show tissue-specific response in different tissues during transportation.     

ORIGINAL ARTICLE: Application of soybean meal,soy protein concentrate and isolate differing in α‐galactosides content to low‐ and high‐fibre diets in growing turkeys

The aim of this experiment was to investigate the physiological and growth response of young turkeys (up to 8 weeks of age) to dietary replacement of soybean meal (SBM) by soy protein concentrate (PC) or protein isolate (PI). This replacement resulted in a differentiated dietary concentration of α‐galactosides of over 2.5% in the SBM diet, approximately 2% with a mixture SBM and PC, 1% with a PC diet and 0.1% with a PI diet. Each treatment was applied in two ways: with lower (3.5%) or higher (5.3%) dietary crude fibre content, made by supplementation with soybean hulls. The highest and lowest body weight of turkeys was recorded both after the first and second 4‐week half of the study in the PC and PI‐type diets respectively. A gradual withdrawal of α‐galactosides from a diet was accompanied by a decline in ileal tissue mass, ileal viscosity and activity of endogenous maltase (the latter was found to be significant at 4 weeks of age). At the same time, two‐way anova revealed that an elevated level of crude fibre (HF treatment) caused an increase in ileal tissue mass (p < 0.05 after 4 weeks of feeding) as well as a decrease in activity level of intestinal sucrase and maltase. The presence of raffinose family oligosaccharides in a diet, in contrast to dietary crude fibre level, significantly affected the caecal metabolism. The rate of bacterial production of short‐chain fatty acids in the caeca was distinctly diminished by dietary withdrawal of α‐galactosides. In conclusion, the soy protein concentrate, in contrast to the protein isolate preparation, exerted positive effects on the turkeys’ growth and gastrointestinal tract physiology and should be considered as an effective SBM substitute.