Adverse Health Effects of the Long-Term Simultaneous Exposure to Arsenic and Particulate Matter in a Murine Model.

PM2.5 and arsenic are two of the most hazardous substances for humans that coexist worldwide. Independently, they might cause multiple organ damage. However, the combined effect of PM2.5 and arsenic has not been studied. Here, we used an animal model of simultaneous exposure to arsenic and PM2.5. Adult Wistar rats were exposed to PM2.5, As, or PM2.5 + As and their corresponding control groups. After 7, 14, and 28 days of exposure, the animals were euthanized and serum, lungs, kidneys, and hearts were collected. Analysis performed showed high levels of lung inflammation in all experimental groups, with an additive effect in the coexposed group. Besides, we observed cartilaginous metaplasia in the hearts of all exposed animals. The levels of creatine kinase, CK-MB, and lactate dehydrogenase increased in experimental groups. Tissue alterations might be related to oxidative stress through increased GPx and NADPH oxidase activity. The findings of this study suggest that exposure to arsenic, PM2.5, or coexposure induces high levels of oxidative stress, which might be associated with lung inflammation and heart damage. These findings highlight the importance of reducing exposure to these pollutants to protect human health.

Dynamics of Gastrointestinal Activity and Ruminal Absorption of the Methane-Inhibitor, Nitroethane, in Cattle.

Nitroethane is a potent methane-inhibitor for ruminants but little is known regarding simultaneous effects of repeated administration on pre- and post-gastric methane-producing activity and potential absorption and systemic accumulation of nitroethane in ruminants. Intraruminal administration of 120 mg nitroethane/kg body weight per day to Holstein cows (n = 2) over a 4-day period transiently reduced (P < 0.05) methane-producing activity of rumen fluid as much as 3.6-fold while concomitantly increasing (P < 0.05) methane-producing activity of feces by as much as 8.8-fold when compared to pre-treatment measurements. These observations suggest a bacteriostatic effect of nitroethane on ruminal methanogen populations resulting in increased passage of viable methanogens to the lower bovine gut. Ruminal VFA concentrations were also transiently affected by nitroethane administration (P < 0.05) reflecting adaptive changes in the rumen microbial populations. Mean (± SD) nitroethane concentrations in plasma of feedlot steers (n = 6/treatment) administered 80 or 160 mg nitroethane/kg body weight per day over a 7-day period were 0.12 ± 0.1 and 0.41 ± 0.1 µmol/mL 8 h after the initial administration indicating rapid absorption of nitroethane, with concentrations peaking 1 day after initiation of the 80 or 160 mg nitroethane/kg body weight per day treatments (0.38 ± 0.1 and 1.14 ± 0.1 µmol/mL, respectively). Plasma nitroethane concentrations declined thereafter to 0.25 ± 0.1 and 0.78 ± 0.3 and to 0.18 ± 0.1 and 0.44 ± 0.3 µmol/mL on days 2 and 7 for the 80 or 160 mg nitroethane/kg body weight per day treatment groups, respectively, indicating decreased absorption due to increased ruminal nitroethane degradation or to more rapid excretion of the compound.

In vitro reduction of methane production by 3-nitro-1-propionic acid is dose-dependent1.

Methanogenesis is a metabolic process that allows the rumen ecosystem the ability to maintain the low hydrogen partial pressures needed for proper digestive function. However, rumen methanogenesis is considered to be an inefficient process because it can result in the loss of 4% to 12% of the total energy consumed by the host. Recent studies have shown that some short-chain nitrocompounds such as nitroethane, 2-nitroethanol, 2-nitro-1-propanol, and 3-nitro-1-propionic acid (3NPA) are capable of inhibiting the production of methane during in vitro culture; nevertheless, optimal supplementation doses have yet to be determined. In the present study, in vitro cultures of freshly collected mixed populations of ruminal microbes were supplemented with the naturally occurring nitrocompound, 3NPA, to achieve 0, 3, 6, 9, or 12 mM. Analysis of fermentation products after 24 h of incubation revealed that methane (CH4) production was reduced in a dose-dependent manner by 29% to 96% (P < 0.05) compared with the amount produced by untreated controls (15.03 ± 0.88 µmol mL-1 incubated liquid). Main effects of the supplement were also observed, which resulted in a reduction (P < 0.05) on amounts of total gas and volatile fatty acids (VFA) produced, as well as in an increase of 0.07 to 0.30 µmol mL-1 on rates of 3NPA degradation. Changes in production of metabolites as CH4, hydrogen (H2), VFA, and NH3 indicated that the fermentation efficiency was not compromised dramatically by 3NPA treatment in moderate doses of 6 and 9 mM. Results further revealed that the metabolism of the 3NPA by microbial populations is also dose-dependent. The microbes were able to metabolize more than 75% of the added nitrocompound, with the greatest degradation rates in cultures treated with 9-mM 3NPA. Finally, from a practical standpoint, and considering the magnitude of CH4 reduction, effect on VFA, and percentage of metabolized supplement, the most efficacious dose for 3NPA administration may be between 3 and 9 mM.

Effect of sole or combined administration of nitrate and 3-nitro-1-propionic acid on fermentation and Salmonella survivability in alfalfa-fed rumen cultures in vitro.

Ruminal methanogenesis is a digestive inefficiency resulting in the loss of dietary energy consumed by the host and contributing to environmental methane emission. Nitrate is being investigated as a feed supplement to reduce rumen methane emissions but safety and efficacy concerns persist. To assess potential synergies of co-administering sub-toxic amounts of nitrate and 3-nitro-1-propionate (NPA) on fermentation and Salmonella survivability with an alfalfa-based diet, ruminal microbes were cultured with additions of 8 or 16mM nitrate, 4 or 12mM NPA or their combinations. All treatments decreased methanogenesis compared to untreated controls but volatile fatty acid production and fermentation of hexose were also decreased. Nitrate was converted to nitrite, which accumulated to levels inhibitory to digestion. Salmonella populations were enriched in nitrate only-treated cultures but not in cultures co- or solely treated with NPA. These results reveal a need for dose optimization to safely reduce methane production with forage-based diets.

Effects of feed-supplementation and hide-spray application of two sources of tannins on enteric and hide bacteria of feedlot cattle.

Pathogenic bacteria attached to the hide or shed in the feces of cattle at slaughter can contaminate carcasses intended to be processed for human consumption. Therefore, new pre-harvest interventions are needed to prevent the carriage and excretion of foodborne pathogens in cattle presented to the processing plant. The objectives of this study were to examine the antimicrobial effects of hydrolysable tannin-rich chestnut and condensed tannin-rich mimosa extracts on bacterial indicators of foodborne pathogens when applied as a hide-intervention and as a feed additive to feedlot cattle. Water (control) or solutions (3 % wt/vol) of chestnut- and mimosa-extract treatments were sprayed (25 mL) at the left costal side of each animal to a 1000 cm² area, divided in four equal quadrants. Hide-swabs samples obtained at pre-, 2-min, 8-h, and 24-h post-spray application were cultured to enumerate Escherichia coli/total coliforms and total aerobic plate counts. In a second experiment, diets supplemented without (controls) or with (1.5 % of diet dry matter) chestnut- or mimosa-extracts were fed during a 42-day experimental feeding period. Weekly fecal samples starting on day 0, and rumen fluid obtained on days 0, 7, 21 or 42 were cultured to enumerate E.coli/total coliforms and Campylobacter. Tannin spray application showed no effect of treatment or post-application-time (P > 0.05) on measured bacterial populations, averaging 1.7/1.8, 1.5/1.6 and 1.5/1.7 (log10CFU/cm²) for E. coli/total coliforms, and 4.0, 3.4 and 4.2 (log10CFU/cm²) in total aerobes for control, chestnut and mimosa treatments, respectively. Mean (± SEM) ruminal E. coli and total coliform concentrations (log(10) CFU/mL) were reduced (P < 0.01) in steers fed chestnut-tannins (3.6 and 3.8 ± 0.1) in comparison with the controls (4.1 and 4.2 ± 0.1). Fecal E. coli concentrations were affected by treatment (P< 0.01), showing the highest values (log10 CFU/g) in fecal contents from mimosa-fed steers compared to controls (5.9 versus 5.6 ± 0.1 SEM, respectively). Total coliforms (log CFU/g) showed the highest values (P < 0.01) in feces from chestnut- and mimosa-fed steers (6.0 and 6.1 ± 0.1 respectively) in comparison with controls (5.7 ± 0.1). Fecal Campylobacter concentrations (log10CFU/g) were affected by treatment (P < 0.05), day (P < 0.001) and their interaction (P < 0.01) with the controls having lower concentrations than chestnut- and mimosa-fed steers (0.4, 1.0, and 0.8 ± 0.3, respectively). It was concluded that under our research conditions, tannins were not effective in decreasing measured bacterial populations on beef cattle hides. Additionally, chestnut tannin reduced E. coli and total coliforms within the rumen but the antimicrobial effect was not maintained in the lower gastrointestinal tract. Further research is necessary to elucidate the possible antimicrobial effects of tannins at site-specific locations of the gastrointestinal tract in beef cattle fed high-grain and high-forage diets.

Effects of nitroethane and monensin on ruminal fluid fermentation characteristics and nitrocompound-metabolizing bacterial populations.

Nitroethane is a potent inhibitor of ruminal CH 4 production, a digestive inefficiency resulting in the loss of 2-15% of a ruminant's gross energy intake and an important emission source of this greenhouse gas. To assess the effect of nitroethane on methanogenesis and characterize ruminal adaptation observed with low treatment doses to this inhibitor, ruminal microbes were cultured in vitro with supplements of water (controls), 4.5 and 9 mM nitroethane, and 0.09 mM monensin, with or without 9 mM nitroethane. All treatments decreased CH 4 production >78% compared to controls; however, differential effects of treatments were observed on CO 2, butyrate isobutyrate, and valerate production. Treatments did not affect H 2 accumulation or acetate and propionate production. Most probable numbers of nitrometabolizing bacteria were increased with 4.5 and 9 mM nitroethane compared to numbers recovered from controls or monensin-containing treatments, which may explain ruminal adaptation to lower nitroethane treatments.

Zoonotic bacterial populations, gut fermentation characteristics and methane production in feedlot steers during oral nitroethane treatment and after the feeding of an experimental chlorate product.

Nitroethane inhibits the growth of certain zoonotic pathogens such as Campylobacter and Salmonella spp., foodborne pathogens estimated to cause millions of human infections each year, and enhances the Salmonella- and Escherichia coli-killing effect of an experimental chlorate product being developed as a feed additive to kill these bacteria immediately pre-harvest. Limited studies have shown that nitroethane inhibits ruminal methane production, which represents a loss of 2-12% of the host's gross energy intake and contributes to global warming and destruction of the ozone layer. The present study was conducted to assess the effects of 14-day oral nitroethane administration, 0 (0X), 80 (1X) or 160 (2X)mg nitroethane/kg body weight per day on ruminal and fecal E. coli and Campylobacter, ruminal and fecal methane-producing and nitroethane-reducing activity, whole animal methane emissions, and ruminal and fecal fermentation balance in Holstein steers (n=6 per treatment) averaging 403+/-26 (SD) kg BW. An experimental chlorate product was fed the day following the last nitroethane administration to determine effects on E. coli and Campylobacter. The experimental chlorate product decreased (P<0.001) fecal, but not ruminal (P>0.05) E. coli concentrations by 1000- and 10-fold by 24 and 48 h, respectively, after chlorate feeding when compared to pre-treatment concentrations (>5.7 log(10) colony forming units/g). No effects (P>0.05) of nitroethane or the experimental chlorate product were observed on fecal Campylobacter concentrations; Campylobacter were not recovered from ruminal contents. Nitroethane treatment decreased (P<0.01) ruminal (8.46, 7.91 and 4.74+/-0.78 micromol/g/h) and fecal (3.90, 1.36 and 1.38+/-0.50 micromol/g/h) methane-producing activity for treatments 0X, 1X and 2X, respectively. Administration of nitroethane increased (P<0.001) nitroethane-reducing activity in ruminal, but not fecal samples. Day of study affected ruminal (P<0.0001) but not fecal (P>0.05) methane-producing and nitroethane-reducing activities (P<0.01); treatment by day interactions were not observed (P>0.05). Ruminal accumulations of acetate decreased (P<0.05) in 2X-treated steers when compared with 0X- and 1X-treated steers, but no effect (P>0.05) of nitroethane was observed on propionate, butyrate or the acetate to propionate ratio. Whole animal methane emissions, expressed as L/day or as a proportion of gross energy intake (%GEI), were unaffected by nitroethane treatment (P>0.05), and were not correlated (P>0.05) with ruminal methane-producing activity. These results demonstrate that oral nitroethane administration reduces ruminal methane-producing activity but suggest that a microbial adaptation, likely due to an in situ enrichment of ruminal nitroethane-reducing bacteria, may cause depletion of nitroethane, at least at the 1X administration dose, to concentrations too low to be effective. Further research is warranted to determine if the optimization of dosage of nitroethane or related nitrocompouds can maintain the enteropathogen control and anti-methanogen effect in fed steers.