Effect of cyclodextrin diallyl maleate on methane production, ruminal fermentation and microbes in vitro and in vivo

Effects of β‐cyclodextrin diallyl maleate (CD‐M) on methane production, ruminal fermentation and digestibility were studied both in vitro and in vivo. In in vitro study, diluted ruminal fluid (30 mL) was incubated anaerobically at 38°C for 6 and 24 h with or without CD‐M using hay plus concentrate (1.5:1) as a substrate. The CD‐M was added at different concentrations (0, 1.25, 2.5, 5.0 and 7.5 g/L). The pH of the medium and numbers of protozoa were not affected by the addition of CD‐M. Total volatile fatty acids were increased and ammonia‐N was decreased, molar proportion of acetate was decreased and propionate was increased (P < 0.05) by CD‐M. Methane was inhibited (P < 0.05) by 14–76%. The effect of CD‐M on methane production and ruminal fermentation was further investigated in vivo using four Holstein steers in a cross‐over design. The steers were fed Sudangrass hay and concentrate mixture (1.5:1) with or without CD‐M (2% of feed dry matter) as a supplement. Ruminal proportion of acetate tended to decrease and that of propionate was increased (P < 0.05) 2 h after CD‐M dosing. Total viable counts, cellulolytic, sulfate reducing, acetogenic bacteria and protozoa were unaffected while methanogenic bacteria were decreased (P < 0.05) by CD‐M. The plasma concentration of glucose was increased, whereas that of urea‐N was decreased (P < 0.05). Methane was inhibited (P < 0.05) from 36.4 to 30.1 L/kg dry matter intake by the addition of CD‐M. Apparent digestibilities of dry matter and neutral detergent fiber were not affected while that of crude protein was increased (P < 0.05) in the medicated steers. These data suggested that dietary supplementation of CD‐M decreased methane production and improved nutrient use.     

Effects of cyclodextrin-iodopropane complex on methane production, ruminal fermentation and microbes, digestibility and blood metabolites in steers

The effects when adding cyclodextrin‐iodopropane complex (CD‐IP) to a diet, on ruminal fermentation and microbes, digestibility, blood metabolites and methane production, were evaluated using four Holstein steers in a cross‐over design. The steers were fed Sudangrass hay plus concentrate mixture at a ratio 1.5:1, and CD‐IP (1% of dry matter) was given twice daily by mixing with concentrate mixture. Rumen and blood samples were collected at 0, 2, and 5 h after morning dosing. Ruminal pH and numbers of protozoa were unaffected by CD‐IP treatment. Ruminal molar proportion of acetate was decreased (P < 0.05), and propionate was increased (P < 0.01) at 2 h after CD‐IP dosing. Proportion of butyrate was increased (P < 0.05) and ammonia‐N was decreased (P < 0.05) at 2 and 5 h after CD‐IP dosing. Adding CD‐IP had no effect on the feed intake and digestion of nutrients. Plasma glucose was increased and urea‐N was decreased (P < 0.05) at 2 and 5 h after CD‐IP dosing. Methane production was decreased (P < 0.05) by approximately 18% in the treatment steers. Numbers of methanogenic bacteria were decreased (P < 0.05), while total viable counts, cellulolytic, sulfate reducing and acetogenic bacteria were unaffected. The present results are the first to show that CD‐IP can partially inhibit in vivo ruminal methanogenesis without adverse effects on digestion of nutrients.     

Comparison of N-terminal pro-atrial natriuretic peptide and three cardiac biomarkers for discriminatory ability of clinical stage in dogs with myxomatous mitral valve disease

Plasma N-terminal pro-atrial natriuretic peptide (NT-proANP) concentration increases with progression of myxomatous mitral valve disease (MMVD) in dogs. This multicentre, prospective study compared plasma NT-proANP, N-terminal pro-brain natriuretic peptide (NT-proBNP), ANP, and cardiac troponin I (cTnI) concentrations in dogs with MMVD for their characteristics and discriminatory ability to detect cardiac dilatation and congestive heart failure (CHF). Thirty-six healthy dogs and 69 dogs with MMVD were included. Clinical variables were obtained via physical examination, thoracic radiography, and echocardiography. The discriminatory ability of each cardiac biomarker (CB) to determine the presence or absence of cardiac dilatation (event 1) and CHF (event 2) was evaluated using the receiver operating characteristic curves. Plasma NT-proANP, NT-proBNP, and ANP concentrations showed a significant association with the left atrium/aorta ratio (P<0.01). The area under the curve of plasma NT-proANP and NT-proBNP concentrations were 0.72 and 0.75, respectively in event1 and 0.72 and 0.76, respectively in event2. Plasma NT-proANP and NT-proBNP concentrations showed sensitivity 80.0 and 80.0%; specificity 67.6 and 64.7% in event1 (cutoff value; 8,497.81 pg/ml and 1,453.00 pmol/l, respectively) and sensitivity 85.7 and 81.0%; specificity 60.4 and 64.6% in event2 (cutoff value; 8,684.33 pg/ml and 1,772.00 pmol/l, respectively). In dogs with MMVD, plasma NT-proANP, NT-proBNP, and ANP concentrations increase with left atrial enlargement. Particularly, plasma NT-proANP and NT-proBNP concentrations appeared to be equally useful in the discriminatory ability to detect cardiac dilatation and CHF.     

The N-Terminal Region of the Readthrough Domain Is Closely Related to Aphid Vector Specificity of Soybean dwarf virus

ABSTRACT It has been speculated that the N-terminal half of the readthrough domain (RTD) encoded by open reading frame 5 of Soybean dwarf virus (SbDV) is related to the vector specificity. To further investigate this hypothesis, transmissibility via aphids was tested on 17 SbDV isolates and comparisons of the deduced amino acid sequences of the coat protein (CP) and other proteins encoded by the RTD were made between these isolates. Isolates were distinguished into four strains: YS, causing yellowing in soybean and transmittable by Aulacorthum solani; DS, causing dwarfing and transmittable by A. solani; YP, causing yellowing and transmittable by Acyrthosiphon pisum; and DP, causing dwarfing and transmittable by A. pisum. Phylogenetic analysis showed that the trees for the CP and the C-terminal half of the RTD sequences contained clusters of isolates of the same symptom type, whereas the tree for the N-terminal half of the RTD contained clusters of isolates of the same aphid vector type. These results agreed with our previous data of the complete nucleotide sequences of four SbDV isolates, and strongly indicated a close relationship between the N-terminal half of the RTD amino acid sequences and aphid transmission specificity of SbDV.     

Bovine herpesvirus-1 (BHV-1) recombinant expressing pseudorabies virus (PrV) glycoproteins B and C induces type 1 immune response in BALB/c mice

Bovine herpesvirus 1 (BHV-1) attached poorly and penetrated into a mouse cell line, BALB 3T3/A31, but a recombinant BHV-1/TF7-6, which expresses pseudorabies virus (PrV) gB and gC genes, did attach and penetrated into cells more efficiently. In this study the gene green fluorescent protein (GFP) has been integrated into genome of BHV-1/TF7-6 and its parental line of BHV-1. When the mouse mesenteries were incubated in vitro and infected with BHV-1/TF7-6/GFP, strong fluorescence was observed while BHV-1/GFP infection hardly demonstrated fluorescence, suggesting that BHV-1 recombinant expressing PrV gB and gC can infect mouse tissue cells more efficiently than the parental BHV-1 does. When BALB/c mice were inoculated with purified BHV-1/TF7-6 or its parental BHV-1, the former induced lower level of anti-BHV-1 immunoglobulin G (IgG) than the latter did. When sub-classes of anti-BHV-1 IgG were analyzed, it was found that mice immunized with BHV-1/TF7-6 or the parental BHV-1 demonstrated the same level of IgG2a. Since anti-BHV-1 IgG1 level was lower in mice inoculated with BHV-1/TF7-6, the IgG2a:IgG1 ratio was higher in BHV-1/TF7-6 inoculated mice than in the parental BHV-1 inoculated ones. These results indicate that BHV-1/TF7-6 induces type 1 predominant immune to BALB/c mice.     

Adrenocorticotropic hormone-induced secretion of cortisol in goats is inhibited by androgen

In a previous study, it was found that there are sex differences in goats with respect to the levels of cortisol secretion induced by transportation stress. We also found that treatment of castrated male goats with dihydrotestosterone (DHT) suppressed the increase in plasma cortisol concentration following transportation, but did not suppress the secretion of adrenocorticotropic hormone (ACTH). This suggests that androgen might block ACTH ‐ induced cortisol secretion. In order to examine this hypothesis, the effects of androgen on ACTH‐induced cortisol secretion in goats were investigated. First, castrated male goats were treated with testosterone (T), DHT or cholesterol (cho) for 21–25 days. Cho was used as a control for T and DHT treatment. Then, plasma cortisol concentrations were compared among the hormonal treatments after ACTH injection. Subsequently, the distribution of androgen receptors in the caprine adrenal gland was investigated. There were no differences in the basal cortisol concentrations among the hormonal treatments. However, plasma cortisol concentrations after ACTH injection were significantly lower in T ‐ and DHT ‐ treated goats than in cho ‐ treated goats. Androgen receptors were present in 60% of the cells in the zonae fasciculata and reticularis of the adrenal cortex, the regions that secrete glucocorticoids. These results suggest that androgen may act directly on the adrenal cortex to suppress cortisol secretion induced by ACTH.     

断奶后多系统衰竭综合征病猪中存在沙门氏菌和猪繁殖与呼吸系统综合征病毒共感染

调查了曾经暴发沙门氏菌病的四个猪场的14头病猪,发现在这些猪的肿胀淋巴结中出现带有淋巴细胞排空的肉芽肿性炎症。应用免疫标记和PCR方法在病变部位检测到猪圆环病毒2(PCV2)抗原和PCV2DNA。此外,在这些病猪的肺脏中检测出猪繁殖与呼吸综合征病毒(PRRSV),分离到猪霍乱沙门氏菌。在9头沙门氏菌感染猪中,有5头为沙门氏菌、PMWS与PRRSV并发感染,其数量(55.6%)远远高于沙门氏菌与PMWS感染猪(22.2%)或沙门氏菌与PRRSV感染猪(22.2%)。     

TGF-β and TNF-α stimulate MMP-9 production in murine epidermal keratinocytes

Matrix metalloproteinases 2 and 9 (MMP‐2 and ‐9) are zinc‐dependent metalloenzymes and have gelatin‐degrading activity. Both MMP are known to be secreted by many types of cells and play important roles in several biological changes including tissue remodeling and wound healing. In the present study, a primary culture of murine epidermal keratinocytes was prepared and effects of transforming growth factor‐β (TGF‐β), tumor necrosis factor‐α (TNF‐α) and interferon‐γ (IFN‐γ) on expression of MMP‐2 and MMP‐9 by the keratinocytes was examined. Gelatin zymography revealed that murine epidermal keratinocytes secreted proenzyme forms of MMP‐2 and MMP‐9, but the active forms of both MMP were hardly detectable, indicating that in vitro autoactivation of these proenzymes did not occur. Both TGF‐β and TNF‐α stimulated MMP‐9 production in a dose‐dependent manner, but the MMP‐2 level was not changed. Interferon‐γ hardly affected production of MMP‐2 or MMP‐9. Ribonuclease protection assay demonstrated that TNF‐α increased the level of MMP‐9 mRNA 6‐fold compared to the control, whereas TGF‐β slightly up‐regulated it. These results suggest that expression of MMP‐9 could be regulated by several cytokines in murine epidermal keratinocytes.