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PubMed1992

Effect of guar, pectin, psyllium, soy polysaccharide, and cellulose on breath hydrogen and methane in healthy subjects.

Different types of dietary fiber are fermented to various extents in vitro, but little is known about the effects of fiber on breath hydrogen and methane levels in vivo. Therefore, we studied the effects on breath hydrogen and methane of 15 g of guar, pectin, psyllium, soy polysaccharide, or cellulose in eight healthy subjects over a 12-h period. None of the fibers had a significant effect on breath hydrogen or methane concentrations, compared with the control (fasting). The four methane producers had lower breath hydrogen levels than the nonproducers 1 h after 15 g of lactulose (3 +/- 1 vs. 42 +/- 9, p less than 0.005) and 5-12 h after the different fibers (3.3 vs. 4.8 ppm; pooled SEM = 0.8; p less than 0.025). When the methane responses of the methane producers were expressed as increments relative to the control, there were small differences between treatments, with guar producing a larger response, 8.2 +/- 3.3 ppm, than cellulose, -2.9 +/- 2.3 ppm (p less than 0.05). The incremental methane responses of the different fibers in vivo were related to the previously reported production of propionic acid (r = 0.94, n = 5, p less than 0.02) and methane (r = 0.93, n = 4, NS) from in vitro fermentation of the same fibers. We conclude that methane producers have lower breath hydrogen levels than nonproducers. Purified fermentable and nonfermentable dietary fibers have no effect on breath hydrogen levels over 12 h in subjects previously consuming a normal diet. However, fermentable fibers may produce small increases in breath methane in methane-producing subjects.

Adult↗

PubMed1990

Mechanism of Methane Transport from the Rhizosphere to the Atmosphere through Rice Plants.

To clarify the mechanisms of methane transport from the rhizosphere into the atmosphere through rice plants (Oryza sativa L.), the methane emission rate was measured from a shoot whose roots had been kept in a culture solution with a high methane concentration or exposed to methane gas in the gas phase by using a cylindrical chamber. No clear correlation was observed between change in the transpiration rate and that in the methane emission rate. Methane was mostly released from the culm, which is an aggregation of leaf sheaths, but not from the leaf blade. Micropores which are different from stomata were newly found at the abaxial epidermis of the leaf sheath by scanning electron microscopy. The measured methane emission rate was much higher than the calculated methane emission rate that would result from transpiration and the methane concentration in the culture solution. Rice roots could absorb methane gas in the gas phase without water uptake. These results suggest that methane dissolved in the soil water surrounding the roots diffuses into the cell-wall water of the root cells, gasifies in the root cortex, and then is mostly released through the micropores in the leaf sheaths.

Journal Article↗

PubMed1996

In Situ Analyses of Methane Oxidation Associated with the Roots and Rhizomes of a Bur Reed, Sparganium eurycarpum, in a Maine Wetland.

Methane oxidation associated with the belowground tissues of a common aquatic macrophyte, the burweed Sparganium eurycarpum, was assayed in situ by a chamber technique with acetylene or methyl fluoride as a methanotrophic inhibitor at a headspace concentration of 3 to 4%. Acetylene and methyl fluoride inhibited both methane oxidation and peat methanogenesis. However, inhibition of methanogenesis resulted in no obvious short-term effect on methane fluxes. Since neither inhibitor adversely affected plant metabolism and both inhibited methanotrophy equally well, acetylene was employed for routine assays because of its low cost and ease of use. Root-associated methanotrophy consumed a variable but significant fraction of the total potential methane flux; values varied between 1 and 58% (mean (plusmn) standard deviation, 27.0% (plusmn) 6.0%) with no consistent temporal or spatial pattern during late summer. The absolute amount of methane oxidized was not correlated with the total potential methane flux; this suggested that parameters other than methane availability (e.g., oxygen availability) controlled the rates of methane oxidation. Estimates of diffusive methane flux and oxidation at the peat surface indicated that methane emission occurred primarily through aboveground plant tissues; the absolute magnitude of methane oxidation was also greater in association with roots than at the peat surface. However, the relative extent of oxidation was greater at the latter locus.

Journal Article↗

PubMed2026

Validation of a national genetic evaluation for methane emission in Holstein cattle.

Lactanet Canada launched a genomic evaluation for methane efficiency for Holsteins in April 2023, utilizing milk mid-infrared-predicted methane (CH4) emissions (CH4MIR) as a proxy. This study validated the methane efficiency genomic evaluation using genotyped cows with CH4MIR and CH4 records from GreenFeed systems (CH4GF), along with relative breeding values (RBV) for methane production and methane efficiency from the April 2023 evaluation. In Lactanet's methane efficiency evaluation, a higher RBV indicates more desirable, lower-emitting animals. For the validation, RBV were categorized into quintiles for the CH4MIR dataset and tertiles for the CH4GF dataset to evaluate trends across the RBV distribution. Mean CH4MIR decreased progressively across RBV quintiles for both traits, with all pairwise comparisons among quintiles significantly different. Similarly, CH4GF emissions declined across RBV tertiles, with significant differences observed between the lowest and highest tertiles. Additional analyses using RBV threshold categories confirmed that cows with the highest RBV consistently exhibited lower methane emissions. Linear regression analyses further demonstrated a negative relationship between RBV and methane emissions, supporting the predictive ability of the genomic evaluation. These findings confirm that Canada's genomic evaluation for methane efficiency effectively differentiates cows by methane emission potential, reinforcing its potential as a tool for genetic selection to reduce methane emissions in dairy cattle.

Journal Article↗

PubMed1991

Oxidation of deuterated compounds by high specific activity methane monooxygenase from Methylosinus trichosporium. Mechanistic implications.

Hydrocarbon oxidations catalyzed by methane monooxygenase purified to high specific activity from the type II methanotroph Methylosinus trichosporium OB3b were compared to the same reactions catalyzed by methane monooxygenase from the type I methanotroph Methylococcus capsulatus Bath and liver microsomal cytochrome P-450. The two methane monooxygenases produced nearly identical product distributions, in accord with physical studies of the enzymes which have shown them to be very similar. The products obtained from the oxidation of a series of deuterated substrates by the M. trichosporium methane monooxygenase were very similar to those reported for the same reaction catalyzed by liver microsomal cytochrome P-450, suggesting that the enzymes use similar mechanisms. However, differences in the product distributions and other aspects of the reactions indicated the mechanisms are not identical. Methane monooxygenase epoxidized propene in D2O and d6-propene in H2O without exchange of substrate protons or deuterons with solvent, in contrast to cytochrome P-450 (Groves, J. T., Avaria-Neisser, G. E., Fish, K. M., Imachi, M., and Kuczkowski, R. L. (1986) J. Am. Chem. Soc. 108, 3837-3838), suggesting that the mechanism of epoxidation of olefins by methane monooxygenase differs at least in part from that of cytochrome P-450. Hydroxylation of alkanes by methane monooxygenase revealed close similarities to hydroxylations by cytochrome P-450. Allylic hydroxylation of 3,3,6,6-d4-cyclohexene occurred with approximately 20% allylic rearrangement in the case of methane monooxygenase, whereas 33% was reported for this reaction catalyzed by cytochrome P-450 (Groves, J. T., and Subramanian, D. V. (1984) J. Am. Chem. Soc. 106, 2177-2181). Similarly, hydroxylation of exo,exo,exo,exo-2,3,5,6-d4-norbornane by methane monooxygenase occurred with epimerization, but to a lesser extent than reported for cytochrome P-450 (Groves, J. T., McClusky, G. A., White, R. E., and Coon, M. J. (1978) Biochem. Biophys. Res. Commun. 81, 154-160). A large intramolecular isotope effect, kH,exo/kD,exo greater than or equal to 5.5, was calculated for this reaction. However, the intermolecular kinetic isotope effect on Vm for methane oxidation was small, suggesting that steps other than C-H bond breakage were rate limiting in the overall enzymatic reaction. Similar isotope effects have been observed for cytochrome P-450. These observations indicate a stepwise mechanism of hydroxylation for methane monooxygenase analogous to that proposed for cytochrome P-450.(ABSTRACT TRUNCATED AT 400 WORDS)

Cytochrome P-450 Enzyme System↗

PubMed2005

Methane mass balance at three landfill sites: what is the efficiency of capture by gas collection systems?

Many developed countries have targeted landfill methane recovery among greenhouse gas mitigation strategies, since methane is the second most important greenhouse gas after carbon dioxide. Major questions remain with respect to actual methane production rates in field settings and the relative mass of methane that is recovered, emitted, oxidized by methanotrophic bacteria, laterally migrated, or temporarily stored within the landfill volume. This paper presents the results of extensive field campaigns at three landfill sites to elucidate the total methane balance and provide field measurements to quantify these pathways. We assessed the overall methane mass balance in field cells with a variety of designs, cover materials, and gas management strategies. Sites included different cell configurations, including temporary clay cover, final clay cover, geosynthetic clay liners, and geomembrane composite covers, and cells with and without gas collection systems. Methane emission rates ranged from -2.2 to >10,000 mg CH(4) m(-2) d(-1). Total methane oxidation rates ranged from 4% to 50% of the methane flux through the cover at sites with positive emissions. Oxidation of atmospheric methane was occurring in vegetated soils above a geomembrane. The results of these studies were used as the basis for guidelines by the French environment agency (ADEME) for default values for percent recovery: 35% for an operating cell with an active landfill gas (LFG) recovery system, 65% for a temporary covered cell with an active LFG recovery system, 85% for a cell with clay final cover and active LFG recovery, and 90% for a cell with a geomembrane final cover and active LFG recovery.

Air Pollutants↗

PubMed2002

Anaerobic methane oxidation in a landfill-leachate plume.

The alluvial aquifer adjacent to Norman Landfill, OK, provides an excellent natural laboratory for the study of anaerobic processes impacting landfill-leachate contaminated aquifers. We collected groundwaters from a transect of seven multilevel wells ranging in depth from 1.3 to 11 m that were oriented parallel to the flow path. The center of the leachate plume was characterized by (1) high alkalinity and elevated concentrations of total dissolved organic carbon, reduced iron, and methane, and (2) negligible oxygen, nitrate, and sulfate concentrations. Methane concentrations and stable carbon isotope (delta13C) values suggest anaerobic methane oxidation was occurring within the plume and at its margins. Methane delta13C values increased from about -54 per thousand near the source to > -10 per thousand downgradient and at the plume margins. The isotopic fractionation associated with this methane oxidation was -13.6+/-1.0 per thousand. Methane 13C enrichment indicated that 80-90% of the original landfill methane was oxidized over the 210-m transect. First-order rate constants ranged from 0.06 to 0.23 per year, and oxidation rates ranged from 18 to 230 microM/y. Overall, hydrochemical data suggest that a sulfate reducer-methanogen consortium may mediate this methane oxidation. These results demonstrate that natural attenuation through anaerobic methane oxidation can be an important sink for landfill methane in aquifer systems.

Bacteria, Anaerobic↗

PubMed2005

Management of tropospheric ozone by reducing methane emissions.

Background concentrations of tropospheric ozone are increasing and are sensitive to methane emissions, yet methane mitigation is currently considered only for climate change. Methane control is shown here to be viable for ozone management. Identified global abatement measures can reduce approximately 10% of anthropogenic methane emissions at a cost-savings, decreasing surface ozone by 0.4-0.7 ppb. Methane controls produce ozone reductions that are widespread globally and are realized gradually (approximately 12 yr). In contrast, controls on nitrogen oxides (NOx) and nonmethane volatile organic compounds (NMVOCs) target high-ozone episodes in polluted regions and affect ozone rapidly but have a smaller climate benefit. A coarse estimate of the monetized global benefits of ozone reductions for agriculture, forestry, and human health (neglecting ozone mortality) justifies reducing approximately 17% of global anthropogenic methane emissions. If implemented, these controls would decrease ozone by -1 ppb and radiative forcing by approximately 0.12 W m(-2). We also find that climate-motivated methane reductions have air quality-related ancillary benefits comparable to those for CO2. Air quality planning should consider reducing methane emissions alongside NOx and NMVOCs, and because the benefits of methane controls are shared internationally, industrialized nations should consider emphasizing methane in the further development of climate change or ozone policies.

Agriculture↗

PubMed2006

Contribution of anthropogenic and natural sources to atmospheric methane variability.

Methane is an important greenhouse gas, and its atmospheric concentration has nearly tripled since pre-industrial times. The growth rate of atmospheric methane is determined by the balance between surface emissions and photochemical destruction by the hydroxyl radical, the major atmospheric oxidant. Remarkably, this growth rate has decreased markedly since the early 1990s, and the level of methane has remained relatively constant since 1999, leading to a downward revision of its projected influence on global temperatures. Large fluctuations in the growth rate of atmospheric methane are also observed from one year to the next, but their causes remain uncertain. Here we quantify the processes that controlled variations in methane emissions between 1984 and 2003 using an inversion model of atmospheric transport and chemistry. Our results indicate that wetland emissions dominated the inter-annual variability of methane sources, whereas fire emissions played a smaller role, except during the 1997-1998 El Niño event. These top-down estimates of changes in wetland and fire emissions are in good agreement with independent estimates based on remote sensing information and biogeochemical models. On longer timescales, our results show that the decrease in atmospheric methane growth during the 1990s was caused by a decline in anthropogenic emissions. Since 1999, however, they indicate that anthropogenic emissions of methane have risen again. The effect of this increase on the growth rate of atmospheric methane has been masked by a coincident decrease in wetland emissions, but atmospheric methane levels may increase in the near future if wetland emissions return to their mean 1990s levels.

Atmosphere↗

PubMed2002

In vitro demonstration of anaerobic oxidation of methane coupled to sulphate reduction in sediment from a marine gas hydrate area.

Anaerobic oxidation of methane (AOM) and sulphate reduction were examined in sediment samples from a marine gas hydrate area (Hydrate Ridge, NE Pacific). The sediment contained high numbers of microbial consortia consisting of organisms that affiliate with methanogenic archaea and with sulphate-reducing bacteria. Sediment samples incubated under strictly anoxic conditions in defined mineral medium (salinity as in seawater) produced sulphide from sulphate if methane was added as the sole organic substrate. No sulphide production occurred in control experiments without methane. Methane-dependent sulphide production was fastest between 4 degree C and 16 degree C, the average rate with 0.1 MPa (approximately 1 atm) methane being 2.5 micro mol sulphide day(-1) and (g dry mass sediment)(-1). An increase of the methane pressure to 1.1 MPa (approximately 11 atm) resulted in a four to fivefold increase of the sulphide production rate. Quantitative measurements using a special anoxic incubation device without gas phase revealed continuous consumption of dissolved methane (from initially 3.2 to 0.7 mM) with simultaneous production of sulphide at a molar ratio of nearly 1:1. To test the response of the indigenous community to possible intermediates of AOM, molecular hydrogen, formate, acetate or methanol were added in the absence of methane; however, sulphide production from sulphate with any of these compounds was much slower than with methane. In the presence of methane, such additions neither stimulated nor inhibited sulphate reduction. Hence, the experiments did not provide evidence for one of these compounds acting as a free extracellular intermediate (intercellular shuttle) during AOM by the presently investigated consortia.

Anaerobiosis↗

PubMed2006

Global health benefits of mitigating ozone pollution with methane emission controls.

Methane (CH(4)) contributes to the growing global background concentration of tropospheric ozone (O(3)), an air pollutant associated with premature mortality. Methane and ozone are also important greenhouse gases. Reducing methane emissions therefore decreases surface ozone everywhere while slowing climate warming, but although methane mitigation has been considered to address climate change, it has not for air quality. Here we show that global decreases in surface ozone concentrations, due to methane mitigation, result in substantial and widespread decreases in premature human mortality. Reducing global anthropogenic methane emissions by 20% beginning in 2010 would decrease the average daily maximum 8-h surface ozone by approximately 1 part per billion by volume globally. By using epidemiologic ozone-mortality relationships, this ozone reduction is estimated to prevent approximately 30,000 premature all-cause mortalities globally in 2030, and approximately 370,000 between 2010 and 2030. If only cardiovascular and respiratory mortalities are considered, approximately 17,000 global mortalities can be avoided in 2030. The marginal cost-effectiveness of this 20% methane reduction is estimated to be approximately 420,000 US dollars per avoided mortality. If avoided mortalities are valued at 1 US dollars million each, the benefit is approximately 240 US dollars per tone of CH(4) ( approximately 12 US dollars per tone of CO(2) equivalent), which exceeds the marginal cost of the methane reduction. These estimated air pollution ancillary benefits of climate-motivated methane emission reductions are comparable with those estimated previously for CO(2). Methane mitigation offers a unique opportunity to improve air quality globally and can be a cost-effective component of international ozone management, bringing multiple benefits for air quality, public health, agriculture, climate, and energy.

Air Pollutants↗

PubMed2006

Hydrogen and methane gases are frequently detected in the stomach.

AIM: To investigate the incidence of bacterial overgrowth in the stomach by using a new endoscopic method in which intragastric hydrogen and methane gases are collected and analyzed. METHODS: Studies were performed in 490 consecutive patients undergoing esophagogastroscopy. At endoscopy, we intubated the stomach without inflation by air, and 20 mL of intragastric gas was collected through the biopsy channel using a 30 mL syringe. Intragastric hydrogen and methane concentrations were immediately measured by gaschromatography. H pylori infection was also determined by serology. RESULTS: Most of intragastric hydrogen and methane levels were less than 15 ppm (parts per million). The median hydrogen and methane values (interquartile range) were 3 (1-8) ppm and 2 (1-5) ppm, respectively. The high hydrogen and methane levels for indication of fermentation were decided if the patient had the values more than 90 percentile range in each sample. When a patient had a high level of hydrogen or methane in one or more samples, the patient was considered to have fermentation. The overall incidence of intragastric fermentation was 15.4% (73/473). Intragastric methane levels were higher in the postoperative group than in other groups. None of the mean hydrogen or methane values was related to H pylori infection. CONCLUSION: Hydrogen and methane gases are more frequently detected in the stomach than expected, regardless of the presence of abdominal symptoms. Previous gastric surgery influences on the growth of methane-producing bacteria in the fasting stomach.

Adult↗

PubMed2006

Alkane bromination revisited: "reproportionation" in gas-phase methane bromination leads to higher selectivity for CH3Br at moderate temperatures.

The reaction of methane and bromine is a mildly exothermic and exergonic example of free radical alkane activation. We show here that the reaction of methane and bromine (CH4:Br2 > or = 1) may yield either a kinetically or a thermodynamically determined bromomethane product distribution and proceeds in two main phases between 450 and 550 degrees C under ambient pressure on the laboratory time scale. This is in contrast to the highly exothermic methane fluorination or chlorination reactions, which give kinetic product distributions, and to the endergonic iodination of methane, which yields an equilibrium distribution of iodomethanes. The first phase of reaction between methane and bromine is a relatively rapid consumption of bromine to yield a kinetic methane bromination product distribution characterized by low methane conversion, low methyl bromide selectivity, and higher polybromomethane selectivity. In the second slower phase CHxBr(4-x) reproportionation leads to significantly higher methane conversion and higher methyl bromide selectivity. For methane bromination at 525 degrees C, CH4 conversion and CH3Br selectivity reach 73.5% and 69.5%, respectively, after ample (60 s) time for reproportionation. The high selectivity and simple configuration make this pathway an attractive candidate for scale-up in halogen-mediated methane partial oxidation processes.

Journal Article↗

PubMed1990

Toxicity of 1,1,1-Trichloroethane and Trichloroethene on a Mixed Culture of Methane-Oxidizing Bacteria.

The influence of trichloroethene (TCE; 0 to 65 mg/liter) and 1,1,1-trichloroethane (1,1,1-TCA; 0 to 103 mg/liter) on methane consumption of a mixed culture of methane-oxidizing bacteria was studied in laboratory batch experiments. Increasing concentrations of TCE or 1,1,1-TCA resulted in decreasing methane consumption. Methane consumption was totally inhibited at a concentration of 13 mg of TCE per liter, while methane consumption was still observed at the upper studied concentration of 103 mg of 1,1,1-TCA per liter. The inhibition of methane consumption by TCE depended on the initial concentration of methane. A model accounting for competitive inhibition between methane and TCE or 1,1,1-TCA was used to simulate methane consumption at various concentrations of TCE or 1,1,1-TCA. The simulations indicated that competitive inhibition may be the mechanism causing the inhibitory effect of TCE on methane consumption, while this does not seem to be the case for 1,1,1-TCA.

Journal Article↗

PubMed1997

Spatial distribution and inhibition by ammonium of methane oxidation in intertidal freshwater marshes.

In two intertidal marshes, the vertical distribution in the sediment and inhibition by ammonium of methane oxidation were investigated by slurry incubation experiments. The two sites differ in their dominant vegetation type, i.e., reed and bulrush, and in their heights above sea level. The reed site was elevated with respect to the bulrush site, resulting in a lower frequency and duration of flooding and, consequently, a higher potential for methane oxidation. Methane oxidation decreased with depth in the bulrush and reed slurries, although methane oxidation associated with root material from the bulrush plants increased with depth. Reed root material had a limited capacity for methane oxidation and showed no significant increase with depth. Inhibition of methane oxidation by ammonium was observed in all samples and depended on methane and ammonium concentrations. Increasing ammonium concentrations resulted in greater inhibition, and increasing methane concentrations resulted in less. Ammonium concentrations had to exceed methane concentrations by at least 30-fold to become effective for inhibition. This ratio was found only in the surface layer of the sediment. Hence, the ecological relevance for ammonium inhibition of methane oxidation in intertidal marshes is rather limited and is restricted to the surface layer. Nitrate production was restricted to the 0- to 5-cm-depth slurries.

Journal Article↗

PubMed1987

Factors affecting methane production in humans. Gastrointestinal diseases and alterations of colonic flora.

Breath methane was studied in 394 subjects. Among 152 controls, 50.0% produced methane--42.1% of males and 57.9% of females. One hundred sixteen patients with gastrointestinal diseases were studied. Among 32 with Crohn's disease, only 2 (6.1%) produced methane, as well as 16 of 51 ulcerative colitis patients (31.4%) and 11 of 32 patients with the irritable bowel syndrome (34.4%). Breath methane is thus unusual in Crohn's disease. After bowel cleansing for colonoscopy or surgery, 15 of 18 methane producers became nonproducers, whereas after antibiotic treatment, 24 of 30 producers sustained their methane-producing status. After gentamycin and cephazolin therapy, methane production was abolished in three of eight patients. Slight spontaneous variations in methane production were also noticed with two of 23 control subjects, becoming nonproducers on restudy after 10-25 months. Thus gastrointestinal diseases, bowel cleansing and, to a much lesser degree, antibiotic therapy, affect methane production.

Adolescent↗

PubMed2004

Microbiological activities contributing to nitrogen removal with methane: effects of methyl fluoride and tungstate.

When methane (CH(4)) and O(2) are present, nitrogen can be removed from wastewater that does not contain other organic carbon sources. In this study, microbial activities during methane-dependent denitrification (MDD) were investigated by adding inhibitors of methane-oxidation and denitrification. Sludge susceptible to MDD showed methane oxidation activity in the presence of CH(4) and O(2), and denitrification activity with methanol and acetate under anoxic conditions. Methyl fluoride (CH(3)F) is known to inhibit methane oxidation. When CH(3)F was present, MDD did not occur, perhaps because methane oxidation was inhibited. Tungstate (WO(4)(2-)), a known inhibitor of nitrate reduction, also lowered denitrification activity in the sludge, and partly inhibited methane oxidation. When WO(4)(2-) was added to the medium, MDD almost ceased, perhaps because of a synergic inhibitory effect on denitrification and methane oxidation. These results show that both methane oxidation and denitrification contribute to MDD.

Bacteria↗

PubMed2004

Reducing methane emissions in sheep by immunization against rumen methanogens.

This work was conducted to determine if methane emissions from sheep immunized with an anti-methanogen vaccine were significantly lower than methane emissions from non-immunized sheep, to test the effectiveness of two different vaccine formulations (VF) on methane abatement, and to compare methane emissions measured using a closed-circuit respiration chamber and the sulphur-hexafluoride (SF6) tracer technique. Thirty mature wether sheep were randomly allocated to three treatment groups (n = 10). One group received an immunization of adjuvant only on days 0 and 153 (control), a second group received an immunization with a 3-methanogen mix on days 0 and 153 (VF3 + 3), and a third group received an immunization of a 7-methanogen mix on day 0 followed by a 3-methanogen mix on day 153 (VF7 + 3). Four weeks post-secondary immunization, there was a significant 7.7% reduction in methane production per kg dry matter intake in the VF7 + 3 group compared to the controls (P = 0.051). However, methane emissions from sheep immunized with VF7 + 3 were not significantly different when compared to the sheep in the control group (P = 0.883). The average IgG and IgA antibody titres in both plasma and saliva of the VF3 + 3 immunized sheep were four to nine times higher than those immunized with VF7 + 3 (P< 0.001) at both 3 and 6 weeks post-secondary immunization. Data also revealed that SF6 methane estimates were consistently higher than the respiration chamber estimates and that there was no significant correlation between the SF6 methane estimates and the respiration chamber methane estimates (R2 = 0.11).

Adjuvants, Immunologic↗