Rapid estimation of sulfide in rumen and blood with sulfide-specific ion electrode

Government of Ontario, Canada, XIA, Ontario, Canada
Microchemical Journal (Impact Factor: 2.75). 09/1980; 25(3):388-395. DOI: 10.1016/0026-265X(80)90280-5


Free sulfide in rumen preserved with a sulfide antioxidation reducing buffer (SAOB) is determined directly and rapidly with a sulfide ion electrode using a standard addition technique. Acid-labile sulfide in blood preserved in alkaline cadmium acetate is determined by electrode measurement after acid liberation in a Johnson-Nishita apparatus and absorption in 50% SAOB. The sulfide antioxidant reducing buffer SAOB is not recommended for preservation of blood samples because of its desulfuration effect on S-proteins and S-amino acids present in blood.

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    • "A 30-mL ruminal fluid sample was collected for the measurement of ruminal S 2– . Rumen S 2– was measured between 2 and 3 h after sampling according to Khan et al. (1980) with the following modifications. An Orion Dual Star pH/Ion Selective Electrodes (ISE) meter (Thermo Scientific, Mississauga, ON, Canada, catalog number 100–240V) was used to analyze the ruminal S 2– . "
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    ABSTRACT: This study evaluated the effects of dietary S concentration and forage-to-concentrate ratio (F:C) on ruminal fermentation, S metabolism, and short chain fatty acid (SCFA) absorption in beef heifers. Sixteen ruminally cannulated heifers (initial BW 628 ± 48 kg) were used in a randomized complete block design with a 2 × 2 factorial treatment arrangement. Main factors included F:C (4% forage vs. 51% forage, DM basis), and the S concentration which was modified using differing sources of wheat dried distillers grains with solubles (DDGS) to achieve low and high S diets (LS = 0.30% vs. HS = 0.67% S on a DM basis). Elemental S was also added to increase the S content for the HS diets. Serum sulfate concentration from blood, sulfide (S(2-)) and SCFA concentrations from ruminal fluid, hydrogen sulfide (H2S) concentration from the ruminal gas cap, and urinary sulfate concentration were determined. Continuous rumen pH and SCFA (acetate, butyrate, and propionate) absorption were measured. There were no interactions between S concentration and F:C. The F:C did not affect DMI (P = 0.26) or ruminal S metabolite concentrations (P ≥ 0.19), but ruminal pH was lower (P < 0.01) and SCFA absorption was greater (P < 0.01) for low F:C diets. Heifers fed HS diets had less DMI (P < 0.01), but greater ruminal pH (P < 0.01), greater concentrations of ruminal H2S (P < 0.01) and serum sulfate (P < 0.01), and greater urinary sulfate concentration (P < 0.01) and output (P < 0.01) relative to heifers fed LS diets. Ruminal H2S was positively correlated with serum sulfate (r = 0.89; P < 0.01). Ruminal acetate concentration was not affected (P = 0.26) by dietary S concentration. Heifers fed HS diet had lower (P = 0.01) ruminal propionate concentration and tended to have lower (P = 0.06) butyrate concentration than heifers fed LS diet. Ruminal acetate was greater (P = 0.01) and butyrate was less (P < 0.01) with high F:C diet than low F:C diet. Both HS (P = 0.06) and low F:C (P = 0.07) diets tended to reduce urine output. Feeding HS diets reduced SCFA absorption (P < 0.05). In summary, S metabolism in beef heifers was not influenced by the F:C, but HS reduced DMI, inhibited SCFA absorption, and increased urinary S excretion.
    Full-text · Article · Dec 2013 · Journal of Animal Science
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    • "Rumen sulfide. Immediately after collecting rumen liquor, a sub-sample (3.5 ml) was taken for sulfide analysis with a sulfide-specific ion electrode (Model 9616 Sure-Flow Combination Silver/Sulfide Electrode; Thermo Orion, Beverly, MA, USA) connected to a specific ion meter (pH/ISE Meter Model 710A, Thermo Orion, Beverly, MA, USA) following the manufacturer's instructions and the method of Khan et al. (1980) "
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    ABSTRACT: Two metabolism trials (experiments 1 and 2) were conducted to examine the effect of the organic S compound, sodium 3-mercapto-1-propane sulfonic acid (MPS) on feed intake, fiber digestibility, rumen fermentation and abundance of cellulolytic rumen microorganisms in cattle fed low S (<0.11%) roughages. Urea was provided in all treatments to compensate for the N deficiency (<0.6%) in the roughages. In experiment 1, steers (333 ± 9.5 kg liveweight) were fed Angleton grass (Dicanthium aristatum) supplemented with S in equivalent amounts as either MPS (6.0 g/day) or sodium sulfate (9.56 g/day). Supplementation of Angelton grass with either sulfate or MPS resulted in an apparent increase in flow of rumen microbial protein from the rumen. Sulfur supplementation did not significantly change whole tract dry matter digestibility or intake, even though sulfate and MPS supplementation was associated with an increase in the relative abundance of the fibrolytic bacteria Fibrobacter succinogenes and anaerobic rumen fungi. Ruminal sulfide levels were significantly higher in the sulfate treatment, which indicated that the bioavailability of the two S atoms in the MPS molecule may be low in the rumen. Based on this observation, experiment 2 was conducted in which twice the amount of S was provided in the form of MPS (8.0 g/day) compared with sodium sulfate (6.6 g/day) to heifers (275 ± 9 kg liveweight) fed rice straw. Supplementation with MPS compared with sulfate in experiment 2 resulted in an increase in concentration of total volatile fatty acids, and ammonia utilization without a change in feed intake or whole tract fiber digestibility even though S and N were above requirement for growing cattle in both these treatment groups. In conclusion, supplementation of an S deficient low-quality roughage diet with either MPS or sodium sulfate, in conjunction with urea N, improved rumen fermentation, which was reflected in an increase in urinary purine excretion. However, MPS appeared to have a greater effect on stimulating short-chain fatty acid production and ammonia utilization when provided at higher concentrations than sulfate. Thus, the metabolism of MPS in the rumen needs to be investigated further in comparison with inorganic forms of S as it may prove to be more effective in stimulating fermentation of roughage diets.
    Full-text · Article · Jun 2009 · animal
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    • "noted Lower detection limit (μM) Reference Steer blood ISE indirect (METH) 2 N/A [33] Wistar rat blood GC-ECD alk ext dx PFBBr, (METH) N/D N/A 0.3 [29] "
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    ABSTRACT: Hydrogen sulfide (H(2)S) is gaining acceptance as a signaling molecule and has been shown to elicit a variety of biological effects at concentrations between 10 and 1000 micromol/l. Dissolved H(2)S is a weak acid in equilibrium with HS(-) and S(2-) and under physiological conditions these species, collectively referred to as sulfide, exist in the approximate ratio of 20% H(2)S, 80% HS(-) and 0% S(2-). Numerous analyses over the past 8 years have reported plasma or blood sulfide concentrations also in this range, typically between 30 and 300 micromol/l, thus supporting the biological studies. However, there is some question whether or not these concentrations are physiological. First, many of these values have been obtained from indirect methods using relatively harsh chemical conditions. Second, most studies conducted prior to 2000 failed to find blood sulfide in micromolar concentrations while others showed that radiolabeled (35)S-sulfide is rapidly removed from blood and that mammals have a relatively high capacity to metabolize exogenously administered sulfide. Very recent studies using H(2)S gas-sensing electrodes to directly measure sulfide in plasma or blood, or HPLC analysis of head-space gas, have also indicated that sulfide does not circulate at micromolar levels and is rapidly consumed by blood or tissues. Third, micromolar concentrations of sulfide in blood or exhaled air should be, but are not, malodorous. Fourth, estimates of dietary sulfur necessary to sustain micromolar levels of plasma sulfide greatly exceed the daily intake. Collectively, these studies imply that many of the biological effects of sulfide are only achieved at supra-physiological concentrations and they question whether circulating sulfide is a physiologically relevant signaling molecule. This review examines the blood/plasma sulfide measurements that have been reported over the past 30 years from the perspective of the analytical methods used and the potential sources of error.
    Full-text · Article · May 2009 · Biochimica et Biophysica Acta
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