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Access to water along a bird's migratory flyway is essential during the vital process of migration. Because of the scarcity of water in some environments, there is potential for migratory birds to encounter and drink from contaminated bodies of water. Ingestion of contaminated water may cause injury and compromise flying ability, leading to a disruption of migration. To determine injury to birds from potential exposure, it is essential to know not only the concentration of a given contaminant in the water but also the quantity and rate of water consumption by the birds. Homing pigeons (Columba livia) were used in a series of experiments to determine differences in drinking behavior after various flights and after periods of resting. Results from the present study demonstrate that homing pigeons' water consumption is dramatically different when assessed according to activity, flight distance, and time elapsed after flight. This suggests that the drinking rates of birds during migration are extremely important and much greater than estimated using traditional exposure assessment procedures. Thus, exposure to contaminants via drinking water may be greatly underestimated, and the rate of water consumption should be considered when estimating potential exposure risk to avian species. Integr Environ Assess Manag 2017;13:870-876. © 2017 SETAC.

Larvae of the black swallowtail butterfly, Papilio polyxenes, the southern armyworm, Spodoptera eridania, and the cabbage looper, Trichoplusia ni, have different feeding habits and dietary breadth, which contributes to differences in their exposure and tolerance to dietary prooxidant allelochemicals. The antioxidant enzyme activities of larvae of these insects have been previously determined, with the levels being P. polyxenes > S. eridania > T. ni. The relative activities of these antioxidant enzymes are consistent with the relative exposure of these insects to prooxidants. This suggests that the antioxidant enzymes may play a role in the defense against allelochemical toxicity in these insects. Dietary diethlydithiocarbamate (DETC), a copper chelating agent and superoxide dismutase (SOD) inhibitor, was shown to inhibit SOD in all three insects. Toxicological studies were conducted using four diets for each insect. The standard diets for each insect were supplemented with either control (solvent), quercetin (a prooxidant), DETC, or DETC plus quercetin. Nontoxic doses of each compound for each insect were used. Inhibition of SOD in P. polyxenes and S. eridania dramatically increased quercetin-induced toxicity as measured by relative growth and consumption rates in these species. DETC had no effect on quercetin toxicity in T. ni. These results elucidate the important role of SOD in the prooxidant allelochemical defense of insects.

The black swallowtail butterfly, Papilio polyxenes, larvae are specialized feeders of pro-oxidant rich plants of Apiaceae and Rutaceae. An important defense against toxic forms of oxygen species generated by ingestion of the pro-oxidants, are the antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT), GSH-dependent glutathione peroxidases (selenium-dependent glutathione peroxidase [GPOX] and peroxidase activity of selenium-independent glutathione-S-transferase [GTpx]), and glutathione reductase (GR). The subcellular distribution of these enzymes in black swallowtail larvae was investigated and was found to resemble the patterns described for larvae of two other lepidopteran species: the southern armyworm, Spodoptera eridania, and the cabbage looper, Trichoplusia ni. The confinement of SOD in the cytosol and mitochondria was typically eukaryotic, but the relative proportion (1:1) was markedly different from the mammalian pattern (4:1; cytosol:mitochondria). The most obvious difference between the black swallowtail and other lepidoptera as a group, and mammalian species, is in very wide intracellular distributions of CAT, GTpx, and GR in insect species. Insects possess very low levels of a GPOX-like activity which reduces both H2O2 and organic peroxides. Consequently, insects have elaborate activities with a wide subcellular distribution of both CAT which decomposes H2O2, and GTpx which decomposes organic peroxides. The reduction of peroxides is dependent on GSH, which in this process is oxidized to GSSG. GR which reduces GSSG to GSH is also of wide subcellular distribution, analogous to the distribution pattern of GTpx.

In mid-fifth-instar larvae of the southern armyworm, Spodoptera eridania, the subcellular distribution of four antioxidant enzymes—superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPOX), and glutathione reductase (GR)—were examined. Two-thirds (4.26 units ·mg protein−1) of the SOD activity was found in the cytosol, and one-thirds (2.13 units ·mg protein−1) in the mitochondria. CAT activity was unusually high and not restricted to the microsomal fraction where peroxisomes are usually isolated. The activity was distributed as follows: cytosol (163 units) mitochondria (125 units) and microsomes (119 units). Similar to CAT, the subcellular compartmentalization of both GPOX and GR was unusual. No activity was detected in the cytosol, but in mitochondria and microsomes, GR levels were 5.49 and 3.09 units. Although GPOX activity exhibited 14–16-fold enrichment in mitochondria and microsomes, respectively, over the 850g crude homogenate, the level was negligible (mitochondria = 1.4 × 10−3 units; microsomes = 1.6 × 10−3 units), indicating that this enzyme is absent. The unusual distribution of CAT has apparently evolved as an evolutionary answer to the absence of GR from the cytosol, and the lack of GPOX activity.

The black swallowtail butterfly larvae, Papilio polyxenes, are specialist feeders that have adapted to feeding on plants containing high levels of prooxidant allelochemicals. Third, fourth, and fifth instar larvae were tested for their antioxidant enzyme activities, superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), and glutathione peroxidase (GPOX), using 850-g supernatants from whole-body homogenates. The overall antioxidant enzyme profile for P. polyxenes was high compared to other insects, with activities ranging as follows: SOD, 1.1–7.5; CAT, 124–343; GR, 1.0–7.5; and GPOX, 0 units. To determine whether these antioxidant enzymes were inducible, P. poly xenes larvae were given a prooxidant challenge by dipping parsley leaves (their diet in the initial studies) in solutions of quercetin, such that the leaves became coated with this prooxidant flavonoid. Mid-fifth instar larvae fed on quercetin-coated leaves were assayed for antioxidant enzyme activities as was previously done with the larvae fed the standard diet. Food consumption and quercetin intake were monitored. SOD activity was increased almost twofold at the highest quercetin concentration tested. CAT and GR activity, on the other hand, were inhibited by increased quercetin consumption, with GR activity completely inhibited at the highest quercetin concentration after 12 h of feeding. GPOX activity, not present in control insects, was also not inducible by a quercetin challenge. These studies point out the key role that the antioxidant enzymes play in insect defenses against plant prooxidants.

In third-, fourth-, and fifth-instar larvae of the cabbage looper moth, Trichoplusia ni, the activities of the antioxidant enzymes, superoxide dismutase (SOD*), catalase (CAT), glutathione peroxidase (GPOX), and glutathione reductase (GR) were examined using 850 g supernatants of whole-body homogenates. The enzyme activities, expressed as units mg−1 protein min−1 at 25°C ranged as follows: SOD, 0.67-2.13 units; CAT, 180.5-307.5 units; GPOX, none detectable; and GR, 0.40-1.19 units. There was a similar pattern of changes for SOD and CAT activities with larval ontogeny, but not for GR. The cabbage looper apparently uses SOD and CAT to form a “defensive team” effective against endogenously produced superoxide anion (O2⪸). Glutathione may serve as an antioxidant for the destruction of any organic/lipid peroxides formed, and GSH oxidized to glutathione disulfide would be recycled by GR. Bioassays against pro-oxidant compounds exogenous sources of (O2⪸) show high sensitivity of mid-fifth instars to the linear furanocoumarin, 8-methoxypsoralen (xanthotoxin) primarily from photoactivation (320-380 nm), and auto-oxidation of the flavonoid, quercetin. The LC50s are 0.0004 and 0.0045% (w/w) concentration of xanthotoxin and quercetin, respectively. Both pro-oxidants have multiple target sites for lethal action and, in this context, the role of antioxidant enzymes is discussed.