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.     

Hemorrhagic necrotizing splenitis in a slaughter pig infected with Arcanobacterium species

A 6-month-old barrow presented with lethargy, inappetence and dysstasia. At necropsy, multiple coalescing hemorrhagic foci were detected in the margins of the spleen. Gram-positive bacilli were isolated from the spleen, kidney, muscle and liver. Comparative 16S rDNA gene sequencing analysis of the isolates (TO16177) revealed that they would be the same species of unpublished Arcanobacterium species strain HJ57-14E (accession no. gi 18873551) (99.7% similarity based on a comparison of 675 bp). Histologic examination of the splenic tissue sections revealed extensive necrosis and inflammation, and gram-positive bacilli were discernible. Multifocal necrosis was also detected in the liver. Immunohistochemically, the isolates were cross-reacted with polyclonal antibodies against Arcanobacterium pyogenes and Actinomyces naeslundii, and the reaction was strong for the latter. Similar reactions were found in the suppurative lesions of the tonsil, and occasionally in the spleen and lymph nodes. The present results indicate that the unpublished Arcanobacterium species induced multiple organ failure accompanied by acute hemorrhagic necrotizing splenitis in this growing-finishing pig.     

Adsorption of Cr(VI) ion on synthetic hydrated oxides of iron

Abstract

The adsorption of Cr(VI) ion on synthetic hydrated oxides of iron (goethite (α-FeOOH) and lepidocrocite (γ-FeOOH)) with different crystallinities was studied in order to remove the Cr(VI) ion from polluted water. At pH 4.5, the amounts of the adsorbed Cr(V1) ion on goethite and lepidocrocite were 51.9 and 62.2 mmol g^?, respectively. Probably, this is due to the fact that the structure of lepidocrocite is more open than that of goethite. Lepidocrocite, therefore, is more effective for the removal of Cr( V1) ion from Cr(Vl)-contaminated and acidic groundwater and sewage.     

The influence of phosphate fertilizer application levels and cultivars on cadmium uptake by Komatsuna (Brassica rapa L. var. perviridis)

Cadmium (Cd) is a common impurity in phosphate fertilizers and application of phosphate fertilizer may contribute to soil Cd accumulation. Changes in Cd burdens to agricultural soils and the potential for plant Cd accumulation resulting from fertilizer input were investigated in this study. A field experiment was conducted on Haplaquept to investigate the influence of calcium superphosphate on extractable and total soil Cd and on growth and Cd uptake of different Komatsuna (Brassica rapa L. var. perviridis) cultivars. Four cultivars of Komatsuna were grown on the soil and harvested after 60 days. The superphosphate application increased total soil Cd from 2.51 to 2.75?mg?kg^?1, 0.1?mol?L^?1 hydrochloric acid (HCl) extractable Cd from 1.48 to 1.55?mg?kg^?1, 0.01?mol?L^?1 HCl extractable Cd from 0.043 to 0.046?mg?kg^?1 and water extractable Cd from 0.0057 to 0.0077?mg?kg^?1. Cd input reached 5.68?g?ha^–1 at a rate of 240?kg?ha^–1 superphosphate fertilizer application. Superphosphate affected dry-matter yield of leaves to different degrees in each cultivar. ‘Nakamachi’ produced the highest yield in 2008 and ‘Hamami No. 2’ in 2009. Compared with the control (no phosphate fertilizer), application of superphosphate at a rate of 240?kg?ha^–1 increased the Cd concentration in dry leaves by 0.14?mg?kg^?1 in ‘Maruha’, 1.03?mg?kg^?1 in ‘Nakamachi’, 0.63?mg?kg^?1 in ‘SC8-007’ in 2008, and by 0.19?mg?kg^?1 in Maruha’, 0.17?mg?kg^?1 in ‘Hamami No. 2’, while it decreased by 0.27?mg?kg^?1 in ‘Nakamachi’ in 2009. Field experiments in two years demonstrated that applications of different levels of calcium superphosphate did not influence Cd concentration in soil and Komatsuna significantly. However, there was a significant difference in Cd concentration of fresh and dry Komatsuna leaves among four cultivars in 2008 and 2009. The highest Cd concentration was found in the ‘Nakamachi’ cultivar (2.14?mg?kg^?1 in 2008 and 1.91?mg?kg^?1 in 2009). The lowest Cd concentration was observed in the ‘Maruha’ cultivar (1.51?mg?kg^?1?dry weight (DW)) in 2008 and in the ‘Hamami No. 2’ cultivar (1.56?mg?kg^?1?DW) in 2009. A decreasing trend in Cd concentration was found in ‘Nakamachi’, followed by ‘SC8-007’, ‘Hamami No. 2’ and ‘Maruha’ successively. It is necessary to consider a low-uptake cultivar for growing in a Cd polluted soil. In these two years’ results, ‘Maruha’ cultivar was the lowest Cd uptake cultivar compared to the others.     

Infection of a chimeric simian and human immunodeficiency virus with CCR5-specific HIV-1 envelope to Rhesus macaques

Human immunodeficiency virus (HIV) infects lymphocytes and macrophages via CD4 and chemokine receptors. In this study, the infectivity of a chimeric simian and human immunodeficiency virus (SHIV) having a CCR5-specific HIV-1 envelope gene was examined. A SHIV strain termed SHIV-JRFL could enter cells via CD4 with a chemokine receptor CCR5, not CXCR4, and the viral replication was suppressed by recombinant human RANTES, one of beta-chemokines. The intravenous inoculation of SHIV-JRFL into two rhesus macaques resulted in a systemic infection, though it was rather weak. During the early infection, the production of RANTES from Con A-stimulated PBMCs of the infected monkeys increased. These results suggested that beta-chemokine has the potential to limit the infectivity of an R5-type SHIV.     

Flow cytometric analysis of chicken NK activity and its use on the effect of restraint stress

Immune system is organized by the influence of both neural and endocrine systems. NK activity plays an important role in the innate immunity. In this study, we observed the effects of restraint stress on chicken peripheral blood NK activity. Viability of FITC-labeled RP9 was measured with PI after treatment with the effector cells. Chicken peripheral blood CD8alpha+ cells expressed strong cytotoxic activity, in contrast to thrombocytes, while peripheral blood CD3+ CD8alpha+ cells and CD4+ cells had little cytotoxic activity. Con A supernatant enhanced the cytotoxic activity of CD8alpha+ cells. Therefore, it is considered that these cytotoxic activities measured by flow cytometry (FCM) analysis are NK activity. When chickens were exposed to restraint stress, the levels of serum corticosterone increased transiently over a short period of time while the NK activity decreased. The decreased NK activity, however, did not recover to the intact levels for a long time, even once the serum corticosterone levels had recovered. These data indicate that chicken NK activity is able to be measured by flow cytometric analysis and that restraint stress causes severe damage to the chicken NK activity.     

In vitro characterization of the inhibitory effects of ketoconazole on metabolic activities of cytochrome P-450 in canine hepatic microsomes

OBJECTIVE: To evaluate the inhibitory potency of ketoconazole (KTZ) on the metabolic activities of isozymes of cytochrome P-450 (CYP) in dogs. ANIMALS: 4 healthy 1-year-old male Beagles. PROCEDURE: Hepatic microsomes were harvested from 4 dogs after euthanasia. To investigate the effects of KTZ on CYP metabolic activities, 7-ethoxyresorufin, tolbutamide, bufuralol, and midazolam hydrochloride were used as specific substrates for CYP1A1/2, CYP2C21, CYP2D15, and CYP3A12, respectively. The concentrations of metabolites formed by CYP were measured by high-performance liquid chromatography, except for the resorufin concentrations that were measured by a fluorometric method. The reaction velocity-substrate concentration data were analyzed to obtain kinetic variables, including maximum reaction velocity, Michaelis-Menten constant, and inhibitory constant (Ki). RESULTS: KTZ competitively inhibited 7-ethoxyresorufin O-deethylation and midazolam 4-hydroxylation; it noncompetitively inhibited tolbutamide methylhydroxylation. Bufuralol 1'-hydroxylation was inhibited slightly by KTZ. The mean Ki values of KTZ were 10.6+/-6.0, 170+/-2.5, and 0.180+/-0.131 microM for 7-ethoxyresorufin O-deethylation, tolbutamide methylhydroxylation, and midazolam 4-hydroxylation, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs, KTZ at a therapeutic dose may change the pharmacokinetics of CYP3A12 substrates as a result of inhibition of their biotransformation. Furthermore, no influence of KTZ on the pharmacokinetics of CYP1A1/2, CYP2C21, and CYP2D15 substrates are likely. In clinical practice, adverse drug effects may develop when KTZ is administered concomitantly with a drug that is primarily metabolized by CYP3A12.     

Seasonal changes in digestibility, passage rate and rumen fermentation of alfalfa hay in sika deer (Cervus nippon) under restricted feeding

To investigate seasonal variations in the digestive functions of sika deer, five female sika deer were provided with an amount of alfalfa hay cubes equivalent to voluntary food intake during winter. We measured the rate at which the food passed through the digestive tract, digestibility and rumen fermentation during the summer (August), autumn (November), winter (February) and spring (May). Total mean retention time in the digestive tract during summer and autumn was numerically longer than that in winter and spring, but the difference did not reach significance. Organic matter and fiber were less digestible in summer and autumn than in winter and spring (P < 0.05), whereas the digestibility of the dry matter tended to vary with the seasons (P < 0.1). Ruminal pH values seasonally changed (P < 0.01), and were the lowest in autumn. The concentration of ruminal ammonia‐nitrogen differed significantly among the seasons (P < 0.05), increasing in winter and decreasing during spring and summer. The numbers of protozoa changed significantly among the seasons (P < 0.05), being higher in autumn than in winter and spring, and intermediate in summer. The concentration of total volatile fatty acids was not seasonally affected, but the molar percentages of propionic acid and butyric acid significantly changed according to season (P < 0.05 and P < 0.01, respectively), and the ratio of acetic to propionic acid tended to change with the seasons (P < 0.1). The results of this study suggested that the digestive functions in sika deer, fed a commercial diet at a restricted level, differed notably among the seasons and these variations might partially be due to environmental effects.