Frederick N. Bebe

Exposure to low doses of endosulfan and chlorpyrifos modifies endogenous antioxidants in tissues of rats

Two experiments were conducted in male SD rats (225–250 g) to determine changes in the activities of endogenous antioxidants superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and concentrations of glutathione (GSH) in tissues after exposure to low doses of endosulfan and chlorpyrifos using a whole body exposure technique. In both experiments, 6 rats/group were exposed 3 hr/day, 5 days/week for 30 days to: 0 (control), 5, 10, 20, 40 and 60% of LD50 of either pesticide in 50% ethanol; actual concentrations were: endosulfan = 0, 0.5, 1.0, 2.0, 4.0, 6.0 mg/250 g body weight; chlorpyrifos = 0, 1.9, 3.8, 7.6, 15.2, and 22.8 mg/250 g body weight. Endosulfan decreased erythrocyte SOD by 21% in all groups and chlorpyrifos increased SOD by 18% in groups 40 and 60. Liver SOD was 12%–20% lower after endosulfan exposure; lung SOD was altered: endosulfan decreased activity by 21% and 51% and chlorpyrifos by 58 and 75% in the 40 and 60 groups, respectively (P ≤ 0.05). Both pesticides increased plasma GPX activity at lower levels and reduced it by 26% and 19% in groups 40 and 60, respectively (P ≤ 0.05). Liver GPX increased in the 60 group and lung GPX declined between 20% and 38% after endosulfan exposure. GSH in the liver and lung: endosulfan reduced GSH by about 30% at lower levels and increased by 41% or 70% at higher levels; chlorpyrifos decreased GSH by 28–40% in 20 and 60 groups, respectively (P ≤ 0.05). Exposure to low, increasing levels of endosulfan and chlorpyrifos can differentially modify endogenous antioxidants SOD, GPX and GSH, which may lead to the development of oxidative stress in some tissues.

Effect of high dietary zinc on plasma ceruloplasmin and erythrocyte superoxide dismutase activities in Copper-depleted and repleted rats

The effect of moderately high dietary zinc (Zn) on the activities of plasma (PL) ceruloplasmin (CP), and PL and erythrocyte (RBC) copper (Cu), Zn superoxide dismutase (SOD) was determined in weanling rats fed Cu-deficient (DEF; <1 mg Cu/kg), marginal (MAR; 2 mg Cu/kg), or control (CON; 5 mg Cu/kg) copper diets containing normal or high Zn (HZn; 60 mg/kg) for 4 wk and supplemented with oral Cu (CuS; 5 mg/L) in drinking water for 0, 1, 3, or 7 d. PL Cu decreased (67% compared to CON;p≤0.05) in the DEF and increased to control level after 3 d of CuS; increased in the MAR group after 1 d of CuS. HZn reduced overall PL Cu by 27% in all groups, but did not alter the linear increase in PL Cu between 0 and 3 d of Cu S. PL CP activity altered concomitantly with PL Cu levels: The time course of increase in CP activity after 0–3 d of CuS was not influenced by HZn in the diet and CP declined in the DEF group by 92%. There was no correlation between dietary Cu level and PL CP. PL SOD activity decreased by 46% (p≤.05) in the DEF group, increased to control activity after 1 d of CuS and declined slighty after 7 d; MAR diet did not alter PL SOD. HZn diet increased PL SOD activity in all groups by 150%, reduced activity in the DEF and MAR groups by 65 and 37% and delayed the recovery of PL SOD after CuS. RBC SOD declined in the DEF and MAR groups by 56 and 33% (p≤0.05) and did not respond to CuS; HZn diet did not influence RBC SOD activity. These data indicate that moderately high Zn in the diet reduces PL Cu, but not PL CP activity or the recovery of PL Cu or CP activity after oral CuS of Cu-deficient rats, modifies the response of PL SOD to dietary Cu, but does not influence RBC SOD activity.

Pesticides and Essential Minerals Modify Endogenous Antioxidants and Cytochrome P450 in Tissues of Rats

Two experiments were conducted in male Sprague Dawley (SD) rats (175–200 g) to determine changes in the activities of endogenous antioxidants superoxide dismutase (SOD), glutathione peroxidase (GPX), cytochrome P450 (ethoxyresorufin deethylase; EROD) and concentrations of glutathione (GSH) in the blood, liver, and small intestinal mucosa (IM). In both experiments, six rats/group were fed diets based on the AIN-93M diet (Control) or the same modified to contain either 500 mg calcium (Low Ca), 7 mg Zn (Low Zn): 2 mg copper (Low Cu), 60 mg zinc (High Zn) or 12 mg copper (High Cu) in the following combination: Control, LCa/LZn, LCa/LZn/LCu, or HZn/HCu, with and without a pesticide mixture containing acephate, endosulfan, and thiram at 25% LD50 for four or two weeks. Pesticides decreased feed intake and weight gain in all groups by 28%. Erythrocyte SOD was higher than control in the HZn/HCu group and in the LCa/LZn/LCu and HZn/HCu groups with pesticide (P# 0.05). Plasma GPX declined by more than 55% in all the groups with and without pesticides compared to the control. The LCa/LZn/LCu and HZn/HCu diets with and without pesticides reduced GPX in the IM by up to 88%, 40%, and 74%, respectively, than the control. Plasma GSH was about 20% higher than the control in most groups with and without pesticides in the diet. Liver and IM GSH were higher than the control in the HZn/HCu group, whereas IM GSH concentrations were lower than the control in the LCa/LZn and LCa/LZn/LCu groups (P#0.05). All three experimental diets with and without pesticides had a significant effect on liver EROD activity (P#0.05). The results indicate that endogenous antioxidants and EROD were independently modified by dietary zinc and copper levels and pesticides.

Micronuclei frequency in lymphocytes and antioxidants in the blood of traditional limited-resource farm workers exposed to pesticides

Chronic low-level exposure to synthetic pesticides is implicated in many health conditions that result from the induction of oxidative stress, including cytogenetic damage. The objective of this study was to assess the risk of genotoxicity using micronuclei (MN) formation in lymphocytes and to determine changes in blood antioxidants superoxide dismutase (SOD) in erythrocytes (E) and glutathione (GSH) in E and plasma (PL) in farm workers for six months during a growing season. Blood and urine samples were collected once a month for six months (June to November 2003) from farm workers (n = 15) and urban unexposed controls (n = 10). Lymphocytes from blood were separated by density gradient centrifugation using Histopaque and cultured using the standard technique. There was no significant difference in the cytokinesis blocked proliferation index (CBPI) of lymphocytes between the farm workers and the control group, but there was a 76% increase in average MN frequency in lymphocytes of the farm worker group (P ≤ 0.05). In addition, MN frequency peaked during August as compared to the other months and the controls (P ≤ 0.05). An 18% decline was observed in the activity of E-SOD in the farm worker group (P ≤ 0.05). GSH in E and PL were similar in both groups. These data suggest that the farm workers may be at a greater risk of developing genotoxicity due to continued exposure to pesticides, especially during the intensive growing season.

Short- and long-term exposure to low levels of pesticide and flavonoid mixtures modify endogenous antioxidants in tissues of rats

The objective of this study was to determine if interaction between phytochemical mixtures and low level exposure to pesticides would modify tissue endogenous antioxidants in rats. Two experiments were conducted using dietary flavonoid (F) and oral pesticide (P) mixtures (FM, PM) on changes in the activities of superoxide dismutase (SOD), glutathione peroxidase (GPX) and glutathione concentrations (GSH) in male Sprague Dawley rats (150–175 g) fed for 2 or 4 weeks the AIN 93M diet with or without equal amounts of quercetin, rutin, catechin (FM) at 1.0 mM or 5 mM/kg diet, and with/without PM fed orally at 0.1 mL/d/5d/wk (PM = chlorpyrifos, endosulfan, thiram at 25% LD50 in oil). Our data indicate that (compared to corresponding Control groups): (i) While, 2 weeks of feeding reduced liver and small intestinal mucosal (IM) SOD activity in the PFM1 (PM+FM1) group, 4 weeks of feeding increased only liver SOD activity in PM, FM1, FM5 and PFM1 and PFM5 groups; (ii) Liver and IM GSH levels increased in PFM groups after 2 or 4 weeks of exposure; plasma GSH increased in the groups fed FM5 with or without PM; (iii) Liver GPX activity declined in both 2 and 4 week experiments in the FM and PFM groups, respectively. These data suggests that metabolic interaction between FM and PM, and duration of exposure, can modify endogenous antioxidants. Data also underscore the prooxidant properties of flavonoids.

Modulation of tissue trace metal concentrations in weanling rats fed different levels of zinc and exposed to oral lead and cadmium

Abstract The effects of varying dietary zinc (Zn) and oral lead (Pb) and cadmium (Cd) exposure on tissue trace metal concentrations were investigated in weanling rats using a 4 X 3 factorial design. Male SD rats (72, 6/group) were fed diets containing either Zn deficient (

Dermal exposure to pesticides modifies antioxidant enzymes in tissues of rats.

Abstract Superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) activities were determined in rat tissues after dermal exposure to pesticides. Two experiments were conducted in male SD rats, 190–210 g body weight. Acephate (ACP), methamidophos (MAP) and nicotine (NIC) were dissolved either individually or together in 0.25 mL of 50% ethanol, which contained: AP=12.6 or MAP 1.3 or NIC= 9.6 mg; EXP 1 ‐ individual pesticide exposure; 64 rats, 16/group; EXP 2 ‐ mixture of AP+MAP+NIC at levels of IX, 2X, 3X; 48 rats, 12/group; 0.25 mL of solution or ethanol (Controls) was applied to 25 mm2 area of shaved skin 3 times a week. Half the rats were terminated after 4 weeks and the rest after 4 weeks of stopping exposure. Single pesticides decreased erythrocyte (RBC) SOD by 17 % after exposure and in the NIC group after post exposure (P#0.05). Increasing concentrations of AP+MAP+NIC mixture elevated RBC SOD by 22 % in the 2X and 3X groups and CAT by 13 % in the 3X group (P#0.05); post exposure increased RBC SOD by 2–3 fold and CAT activity by 13 % in all 3 groups. Liver GPX increased by 30–40 % and CAT decreased by 12 % in all exposed and post exposed groups (P#0.05). The results suggest that dermal exposure to mixtures of pesticides can selectively induce SOD, CAT and GPX activities in RBC and liver.

Interaction between pesticides and essential metal copper increases the accumulation of copper in the kidneys of rats

Male Sprague-Dawley rats, weighing 175–200 g, six per group were fed AIN 93M diet (CON) or diets containing 500 mg Ca (LCa), 7 mg Zn (LZn), 2 mg Cu (LCu), 60 mg Zn (HZn), or 12 mg Cu (HCu) per kilogram of diet in the following combinations: control (CON), LCa+LZn (LC+Z), LCa+LZn+LCu (LC+Z+C), or HZn+HCu (HZ+C) without or with a pesticides mixture (PM); Endosulfan, Thiram, and Acephate were added to the diets at 25% of LD50/kg. Rats were fed for 2 wk (small intestinal changes) or 4 wk (tissues changes). Plasma Zn was 47% lower than CON in the experimental groups. Plasma Cu and ceruloplasmin concomintantly decreased in the LC+Z+C group and increased with the addition of PM Kidney Cu was 40% lower in LC+Z+C group, than CON and increased by 31% with PM; in the HZ+C+PM group, kidney Cu was 38% higher than the HZ+C group. Mucosal and small intestines Ca declined by 47% in all experimental groups; PM increased Zn in the LC+Z+C and HZ+C groups; PM further decreased intestinal and muocsal Cu retention in the LC+Z+C and HZ+C groups. Data suggest that low levels of PM in the diet can induce Cu accumulation in the kidney when dietary Zn and Cu are low or high.

Effects of encapsulated magnesium on growth and bone mineral concentrations in weanling rats

The effect of magnesium oxide (MgO) encapsulated (E) with a modified cellulose (Ethocel) on growth and bone minerals was assessed in weanling rats. Ethocel encapsulated Mg was added to bread flour at 10 or 30% of RDA/serving (80 mg/100g or 240 mg/100g dry bread). Bread meal, previously supplemented with Mg, was the sole source of Mg in the diet based on AIN93G. Male Sprague-Dawley rats, 26 d old, 8/group, were given either control diet: AIN93G or bread based diets: Mg10E, Mg30E and Mg30N (nonencapsulated) for 4 weeks. Food intake and weight gain of rats fed Mg10E and 30E diets declined 22 and 45%, respectively (P≤0.05). The decline in liver, femur and tibia weights was consistent with decreased food intake and weight gain. Plasma Mg remained unaltered, while plasma Ca concentrations were 14 and 26% lower than the control in the Mg30E and Mg30N groups (P≤0.05). Femur Mg levels were similar to control, but tibia Mg concentrations were reduced by an average of 24% in Mg10E and Mg30E groups and significantly (43%) in the Mg30N group. There was a significant reduction in tibia Ca in the Mg30N group of about 43%; the decline in femur and tibia calcium in the other two bread groups was less significant. The data suggest that encapsulation of MgO did not alter the bioavailability of Mg; it minimized the adverse effects of supplemental Mg on calcium homeostasis.

Meat Goats in Hillside Pastures: Control of Undesirable Plant Species and GPS Collar Determination of Activity Patterns

ABSTRACT Pasture weed control is complicated by the steep hill side topography found in many areas of Kentucky. Meat goats (Capra hircus) are a possible alternative to mechanical or chemical weed control in these pasture situations. A study using 120 boer-cross female goats averaging 44 kg, was conducted in Carter County (2010–2012) to determine the impact of meat goats on undesirable plant species. Additionally, 15 goats chosen at random were fitted with GPS collars for 5 days to determine animal activity. Plant population and canopy density measurements were recorded at the start, middle, and end of the grazing season for two or three years. Goat periodic preference for particular plant species resulted in an increase in growth of other species for the same period or year. Bramble density in the upper canopy (90 cm and above) decreased during the study, particularly in the first year of grazing, but increased in the second year, as goat preference changed (P < 0.05). Overall, grass availability in the middle and lower canopy (45 cm to 90 cm and 0 to 45 cm, respectively) increased during the grazing season in the study (P < 0.05). Distinct activity patterns related to resting, foraging period and locations were observed in goats from GPS measurements. The effectiveness of goats in eliminating undesirable plants species and the use of GPS as tool in determining behavior showed great potential for using meat goats in a vegetation management plan.