ISSN 19950829, Inland Water Biology, 2013, Vol. 6, No. 4, pp. 351–356. © Pleiades Publishing, Ltd., 2013. Original Russian Text © A.I. Aminov, I.L. Golovanova, A.A. Filippov, 2013, published in Biologiya Vnutrennikh Vod, 2013, No. 4, pp. 82–88.
AQUATIC TOXICOLOGY
Effect of the Herbicide Roundup on the Activity of Glycosidases of Invertebrates and Juvenile Fish A. I. Aminov, I. L. Golovanova, and A. A. Filippov Papanin Institute of Biology of Inland Waters, Russian Academy of Sciences, pos. Borok, Yaroslavl oblast, Nekouzskii raion, 152742 Russia email:
[email protected] Received January 26, 2012
Abstract—The effects of in vitro exposure to the herbicide Roundup at concentrations of 0.1–50 µg/L on the activity of maltase and sucrase and the total amylolytic activity in the organism of invertebrates and fish fry have been investigated. Glycosidases in invertebrates are less sensitive to the herbicide than those in juvenile fish. Roundup has a greater inhibitory effect on glycosidase activity in the tissues of actual prey (roach recov ered from pike stomach) than in potential prey (roach captured in the pond). The magnitude and direction of the effects depend on the animal species and the concentration of the toxicant. Keywords: herbicide, Roundup, fish, invertebrates, digestive enzymes, glycosidases DOI: 10.1134/S1995082913040032
INTRODUCTION Roundup is one of the most popular herbicides based on the isopropylamine salt of glyphosate [N(phosphonomethyl) glycine]. It is widely used for the destruction of weeds in fields, drainage canals, irri gation systems, and ponds. The halflife of glyphosate ranges from 30 to 90 days in the soil and from 7 to 14 days in water [17]; microbiota is involved in the destruction of this compound [23]. Due to its high ability to adsorb to suspended particles, the herbicide can spread over long distances, accumulating in the sediment, where it retains activity for a long time. The mechanism of Roundup action involves the inhibition of enzymes of the shikimic acid pathway, preventing the synthesis of the three aromatic amino acids: phe nylalanine, tyrosine, and tryptophan [12, 23]. The herbicide ingested by aquatic organisms becomes involved in the metabolism and causes disturbances of various body functions [12, 22, 25]. The 96h LC50 values for Roundup vary from 2 to 55 mg/L, depend ing on the fish species, life cycle stage, and experimen tal conditions [22], while the value of 96h LC50 for glyphosate (the active ingredient in Roundup) deter mined for carp Cyprinus carpio (L.) is much higher, namely, 620 mg/L [25]. A large amount of data on the toxicity of Roundup for aquatic organisms, including invertebrates and fish, has been accumulated in the recent years [2, 6, 9, 12, 13, 16, 27, 31, 33]. Aquatic animals, fish in partic ular, may be more sensitive to Roundup than mam mals [20]. The main toxic effects of Roundup are asso ciated with the action of glyphosate; nevertheless, a number of studies [17, 23, 26] showed that polyoxy
ethyleneamine, a surfactant present the herbicide, can be much more toxic than the active ingredient. The results of experiments performed on Hydra attenuate Pallas showed that the Roundup toxicity depends on the ratio of active and auxiliary components, as well as on the concentration of the herbicide [14]. There are only few studies addressing the effect of Roundup on the activity of carbohydratecleaving enzymes in the tissues of animals consumed by fish of different ecological groups [5, 10]. Due to the impor tant role of carbohydrates in the energy and plastic metabolism of the organism and the possible involve ment of prey hydrolases in digestion, especially in the autodegradation of prey tissues [8], investigating the effect of Roundup on these enzymes is of considerable interest both for environmental physiology and for practical fishery. The purpose of the present study was to investigate the in vitro effect of Roundup in sublethal concentra tions (0.1–50.0 µg/L) on glycosidase activity in the whole organism of invertebrate animals and juvenile fish. MATERIALS AND METHODS Juvenile freshwater teleost of eight different spe cies, namely, pikeperch Stizostedion lucioperca (L.) (body weight 0.23 ± 0.01 g, body length 2.64 ± 0.06 cm), pike Esox lucius L. (9.64 ± 0.92 g, 11.20 ± 0.31 cm), roach Rutilus rutilus (L.) (0.25 ± 0.01 g, 3.12 ± 0.05 cm), river perch Perca fluviatilis L. (0.63 ± 0.05 g, 4.02 ± 0.09 cm), sprat Clupeonella cultriventris (Nord.) (0.55 ± 0.03 g, 3.72 ± 0.4 cm), common carp Cyprinus carpio (L). (1.33 ± 0.15 g, 4.03 ± 0.14 cm), crucian carp Carassius auratus (L.) (2.11 ± 0.50 g, 3.93 ± 0.27 cm),
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Table 1. The effect of in vitro treatment by the herbicide Roundup on glycosidase activity (µmol/(g min)) in wholebody homogenates of invertebrates Object
Roundup concentration, µg/L 0
0.1
Zooplankton Chironomids Zebra mussel Pond snail
1.31 ± 0.05a,b 5.97 ± 0.07a 2.93 ± 0.06a 7.92 ± 0.30a
1.10 ± 0.16a,c 5.87 ± 0.08a 2.59 ± 0.06b 7.44 ± 0.21a
Zooplankton Chironomids Zebra mussel Pond snaile
1.24 ± 0.01a 4.94 ± 0.04a 2.25 ± 0.13a 3.15 ± 0.02a
1.43 ± 0.02b 5.26 ± 0.07b 2.64 ± 0.13a,b 3.24 ± 0.03a
Zooplankton Chironomids
0.64 ± 0.03a,b 1.93 ± 0.03a
0.80 ± 0.05b,c 2.00 ± 0.04a,b
1 10 Amylolytic activity 1.33 ± 0.16a,b 1.60 ± 0.13b 5.76 ± 0.11a,b 5.55 ± 0.05b,c 2.52 ± 0.05b 3.11 ± 0.07a,d a,b 7.60 ± 0.13 8.32 ± 0.20b Maltase activity 1.21 ± 0.07a 1.46 ± 0.07a,b b,c 5.31 ± 0.10 5.42 ± 0.07b,c a,b 2.31 ± 0.23 2.62 ± 0.14a,b 3.21 ± 0.05a 3.49 ± 0.02b Sucrase activity 0.75 ± 0.13a,b 0.97 ± 0.03c 2.00 ± 0.04a,b 2.13 ± 0.09b
25
50
0.92 ± 0.04c 5.55 ± 0.10b,c 3.52 ± 0.12c 9.12 ± 0.23c
0.86 ± 0.08c 5.44 ± 0.07c 3.33 ± 0.07c,d 9.60 ± 0.28c
1.46 ± 0.06b 5.31 ± 0.04b,c 2.66 ± 0.07b 3.31 ± 0.07a,b
1.58 ± 0.11b 5.55 ± 0.17c 2.51 ± 0.03a,b 3.53 ± 0.14c
0.58 ± 0.06a 2.05 ± 0.02a,b
0.63 ± 0.05a,b 1.97 ± 0.07a,b
Here and in Table 2, different superscripts indicate statistically significant differences between the values in each row (ANOVA, LSDtest), p < 0.05.
and Amur sleeper Perccottus glenii Dyb. (0.82 ± 0.11 g, 3.59 ± 0.10 cm), and invertebrates, namely, crustacean zooplankton (total samples containing representatives of the Dafniiformes, Copepoda, and Ostracoda genera), larvae of the chironomid Chironomus plumosus (L.), and molluscs (zebra mussel Dreissena polymorpha (Pall.) and large pond snail Limnea stagnalis (L.)) were used as research objects in the present study. Fish and inver tebrates were captured in the coastal zone of Rybinsk reservoir in summer and delivered to the laboratory within 1–2 h. Glycosidase activity was assayed in wholebody homogenates of fish and invertebrates. Animals were placed on a glass plate in an ice bath. Prior to homog enization, the largest individuals were minced with scissors (the shell of mollusks was removed before mincing). Pooled samples, including several hundred crustaceans or 10–20 mollusks or juvenile fish, were processed in a glass homogenizer with cooled (2–4°C) Ringer’s solution for coldblooded animals (110 mM NaCl, 1.9 mM KCl, 1.3 mM CaCl2, pH 7.4) added at a ratio of 1 : 9. The original homogenate was further diluted 2–10 fold in Ringer solution. Substrate solu tions (soluble starch, 18 g/L; 50 mM sucrose, and 50 mM maltose) were prepared in Ringer solution of the composition described above. Homogenates were incubated with substrate solution for 30–60 min at 20°C and pH 7.4 with continuous stirring. The activity of maltase (EC 3.2.1.20) was assayed using the Fotoglyukoza (OOO Impakt, Russia) clini cal biochemistry kit. The activity of saccharase (EC 3.2.1.48), as well as amylolytic activity (a charac teristic of the total activity of enzymes capable of hydrolyzing starch, namely, αamylase (EC 3.2.1.1), glucoamylase (EC 3.2.1.3), and maltase) were assessed by analyzing hexose accumulation according to the modified method of Nelson [11]. Enzymatic activity was determined in five replicates at each point with the
background (glucose content in the original homoge nate) taken into account and expressed in micromoles of the reaction product formed per 1 min of incubation of the enzymatically active preparation with the sub strate per 1 g wet tissue weight (µmol/(g min)). Toxicant solutions were prepared using the Roundup commercial herbicide formulation (pro duced and packaged by ZAO Avgust (Russia) under the license of Monsanto Europe S.A. (Belgium)). The chemical is a 36% aqueous solution of glyphosate. The eventual inert ingredients reinforcing the effect of the active component or facilitating the use of the her bicide were not mentioned in the product annotation. The choice of concentrations for the experiment was motivated by the maximum acceptable concentration values set for the water bodies used in fishery (0.001 mg/L) and the 96h LC50 values for fish and daphnia exposed to Roundup (2–168 mg/L) [10, 28]. The homogenates were preincubated with Roundup (an equal amount of Ringer solution was added to the control samples instead of toxicant solution) for 1 h. Roundup concentrations expressed as glyphosate con tent equaled 0.1, 1, 10, 25 and 50 µg/L. The data were processed statistically using Statgraph ics Plus 5.1 and Excel 2003. The results are presented as mean values and errors of the mean (M ± m). The signif icance of the differences was assessed by single analysis (ANOVA, LSDtest) at p = 0.05 [29]. RESULTS Amylolytic activity changes in wholebody homo genates of different invertebrates exposed to Roundup were of opposing character, with the maximum increase of activity (21%) detected in pond snails and the maximum decrease (30% relatively to control) detected in the tissues of crustacean zooplankton (Table 1). The decrease in enzymatic activity was less INLAND WATER BIOLOGY
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Table 2. The effect of in vitro treatment by the herbicide Roundup on glycosidase activity (µmol/(g min)) in wholebody homogenates of juvenile fish Roundup concentration, µg/L
Object 0
0.1 a
1 a
Sprat Common carp Roach Roach* Amur sleeper Crucian carp Perch Pike
0.51 ± 0.03 4.30 ± 0.18a,b 9.07 ± 0.32a 8.27 ± 0.12a 8.12 ± 0.12a 22.3 ± 0.73a,b 4.11 ± 0.07a,b 3.93 ± 0.24a
0.52 ± 0.03 2.63 ± 0.26c 10.0 ± 0.56a,b 6.67 ± 0.34b 7.24 ± 0.12c 17.9 ± 1.38c 3.79 ± 0.32a,b 4.07 ± 0.12a
Sprat Common carp Roach Roach* Amur sleeper Crucian carp Perch Pike Pikeperch
0.76 ± 0.08a,b 2.14 ± 0.04a 0.90 ± 0.18a 0.88 ± 0.12a 8.02 ± 0.16a,b 9.51 ± 0.41a,b 1.33 ± 0.10a 0.23 ± 0.06a 1.23 ± 0.12a
0.82 ± 0.06b 3.26 ± 0.04b,c 1.51 ± 0.14b 0.70 ± 0.04a,b, c 7.60 ± 0.09b,c 9.20 ± 0.35a,b 1.49 ± 0.03a,b 0.29 ± 0.06a,b 1.15 ± 0.08a
Sprat Common carp Roach Roach* Amur sleeper Crucian carp Pike
0.14 ± 0.01a,b 0.20 ± 0.01a,b 0.72 ± 0.04a 0.77 ± 0.05a 2.53 ± 0.08a 4.45 ± 0.11a,b 0.28 ± 0.06b,c
0.16 ± 0.01b 0.20 ± 0.01a,b 0.82 ± 0.35a 1.27 ± 0.07b 2.19 ± 0.09b,c 4.37 ± 0.11a,b 0.39 ± 0.12c
10
Amylolytic activity 0.47 ± 0.03a 0.47 ± 0.02a d 3.58 ± 0.14 4.00 ± 0.16b,d 10.2 ± 0.54a,b 10.8 ± 0.53b 6.40 ± 0.40b 6.47 ± 0.39b 7.52 ± 0.31b,c 7.96 ± 0.16a,b 19.5 ± 0.80b,c 23.5 ± 0.90a 4.27 ± 0.17a,b 3.49 ± 0.45a 4.13 ± 0.17a 5.40 ± 0.19b Maltase activity 0.52 ± 0.14a 0.97 ± 0.15b 3.48 ± 0.10c 3.15 ± 0.15b c,d 2.07 ± 0.25 2.01 ± 0.08c,d 0.80 ± 0.04a,b 0.81 ± 0.11a,b 7.03 ± 0.07d 7.53 ± 0.10c 8.67 ± 0.56a,b 8.27 ± 0.44a 1.71 ± 0.06c,d 1.82 ± 0.03d 0.38 ± 0.08a,b 0.48 ± 0.05a, b 1.21 ± 0.12a 0.94 ± 0.09a,b Sucrase activity 0.15 ± 0.01a,b 0.15 ± 0.01a,b 0.26 ± 0.02a 0.19 ± 0.02b a 0.81 ± 0.20 0.82 ± 0.14a 0.29 ± 0.05c 0.28 ± 0.07c a 2.48 ± 0.09 2.40 ± 0.09a,b 4.24 ± 0.09a 4.64 ± 0.11b,c a,b 0.18 ± 0.05 0.29 ± 0.05c
25
50
0.49 ± 0.03a 4.95 ± 0.19a 11.4 ± 0.78b 6.13 ± 0.63b 8.58 ± 0.15a 21.9 ± 0.33a,b 4.27 ± 0.44a,b 5.53 ± 0.27b
0.46 ± 0.02a 4.34 ± 0.35a,b 11.1 ± 0.37b 5.60 ± 0.39b 8.04 ± 0.16a,b 23.7 ± 0.27a 4.53 ± 0.40b 5.73 ± 0.29b
0.92 ± 0.05b 3.44 ± 0.10c 2.26 ± 0.11d 0.59 ± 0.10b,c 6.69 ± 0.10d 9.19 ± 0.35a,b 1.69 ± 0.04c,d 0.48 ± 0.04a,b 0.75 ± 0.08b
0.76 ± 0.10a,b 3.32 ± 0.15b,c 1.70 ± 0.14b,c 0.52 ± 0.11c 8.25 ± 0.31a 9.77 ± 0.47b 1.57 ± 0.08b,c 0.50 ± 0.06b 1.27 ± 0.10a
0.13 ± 0.01a 0.19 ± 0.02b 1.01 ± 0.11a 0.33 ± 0.06c 2.16 ± 0.07c 4.83 ± 0.15c 0.45 ± 0.03c
0.15 ± 0.01a,b 0.24 ± 0.02a,b 1.05 ± 0.09a 0.18 ± 0.08c 2.60 ± 0.02a 4.67 ± 0.14b,c 0.12 ± 0.01a,b
* Juvenile roach recovered from pike stomach (actual prey).
pronounced in chironomids, being in the range of 7– 12% of the control. Exposure to low concentrations of Roundup (0.1 and 1 µg/L) caused a decrease in amy lolytic activity of zebra mussel homogenates by 12 and 14%, respectively, while exposure to higher concentra tions (25–50 µg/L) resulted in an increase in activity by 14–20% of the control. Roundup exposure caused a significant increase in maltase and sucrase activity (6–27% of control and 7–53% of control, respec tively) in all the invertebrate animals studied. Effects of Roundup on enzyme activity in the wholebody homogenates of juvenile fish varied mark edly between species. Amylolytic activity in pike and roach increased by 19–46% from the control at Roundup concentrations ranging from 10 to 50 µg/L (Table 2). Exposure of carp homogenates to the toxi INLAND WATER BIOLOGY
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cant at low concentrations (0.1 and 1 µg/L) resulted in a 39 and 17% decrease in amylolytic activity, respec tively, while at 25 µg/L the activity increased by 15% of the control. The amylolytic activity of crucian carp and rotan homogenates decreased by 11–20% of the control at the lowest concentration of Roundup. No significant effects were identified in other species stud ied. However, a clear concentrationdependent inhib itory effect was detected for roach retrieved from the pike stomach (actual prey): as the concentration of Roundup increased from 0.1 to 50 µg/L, the amy lolytic activity decreased by 19–32% of the control. Both increases and decreases in maltase activity were detected in samples treated with Roundup. The activity increased 28–152% in roach, perch, pike, and carp and decreased by 6–16% from the control in
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Amur sleeper and pikeperch. No significant changes in enzyme activity were detected in crucian carp. Mal tase activity was significantly reduced in roach extracted from the pike stomach, especially when high concentrations of Roundup were used; the decrease amounted to 8–41% of the control. Sucrase activity was much lower than amylolytic activity and the activity of maltase in all the species studied. Roundup treatment resulted in an increase in sucrase activity in roach by 13–46% of the control. On the contrary, Roundup treatment caused a 57–77% decrease in sucrase activity in roach extracted from pike stomach. Significant decreases in sucrase activity in Amur sleeper homogenates were detected only at concentrations of 0.1 and 50 µg/L (13 and 15% decrease, respectively). Changes in sucrase activity in juvenile pike had opposite directions at different her bicide concentrations, while no significant changes at all were registered in Rounduptreated homogenates of sprat, crucian carp, and common carp. DISCUSSION Changes in the morphological, physiological, and biochemical parameters in fish and invertebrates acutely or chronically exposed to Roundup have been reported. Sublethal concentrations of Roundup caused histological changes in the liver, gills, and kid neys of Nile tilapia Oreochromis niloticus (L.) [21], as well as in the liver of carp [7, 30] and streaked prochi los Prochilodus lineatus Valenciennes [22]. Moreover, exposure of prochilos to Roundup resulted in an increase in blood glucose level indicative of a typical stress reaction and an increase in liver catalase activity indicative of the activation of the protective antioxi dant system in this fish species [22]. Changes in meta bolic and enzymatic parameters of fish, such as the intensity of protein catabolism and lipid peroxidation (POL), and cholinesterase activity upon exposure to Roundup at concentrations used in aquaculture were demonstrated in South American catfish Rhamdia quelen (Quoy and Gaimard) [19]. Similarly, changes in the activity of enzymes involved in antioxidant pro tection were elicited by Roundup exposure in Prussian carp [23]. Sublethal concentrations of glyphosate (2.5–10 mg/L) caused an increase in the activity of liver alkaline phosphatase [25], a decrease in brain and muscle acetylcholinesterase (AChE) activity, and an increase in lipid peroxidation intensity [12] in the carp. A decrease in acetylcholinesterase activity in the brain was also demonstrated for the headstander Lep orinus obtusidens Valenciennes [18] and prochilos [24]. Glyphosate or polyoxyethyleneamine are supposed to reduce the activity of AChE by interacting with its active site [12]. Glyphosate altered the immune status of Nile tilapia [15] and the behavior of rainbow trout Oncorhynchus mykiss Walbaum [32]. Changes in several biochemical parameters were detected in carp juveniles after chronic exposure to
0.004 mg/L Roundup [6, 7]. Concentrations of several metabolites and the activity of cytoplasmic lactate dehydrogenase and mitochondrial citrate dehydroge nase, as well as liver morphology, were altered, this being indicative of the role of proteins, especially the insoluble fraction, as major energy substrates for the processes of detoxification in twoyear old fish. The total protein content in the liver of carp fingerlings did not change, since the energy for Roundup detoxifica tion during the first seven days of the experiment was derived from carbohydrate catabolism. However, the glucose and glycogen content in the liver did not differ significantly between 1 and 2year old carps after 14 days of exposure to Roundup [6]. The increasing role of proteins in the metabolism was demonstrated in 15day tests on four generations of Daphnia magna Straus exposed to sublethal con centrations of Roundup [10]. The exposure to herbi cide solutions containing 25 and 50 mg glyphosate per liter resulted in an increase in proteolytic activity and a decrease in amylolytic activity in wholebody homo genates of the crustaceans. The magnitude of the effects observed in the second and third generations was half of that observed in the first generation, this being indicative of decreased adaptive capacity. Sucrase activity in crustaceans of the third generation decreased to a greater extent than that in the parent line, revealing a functional accumulation of the toxic effect. Glycosidase activity in daphnia reared in the labo ratory was affected by Roundup exposure to a greater extent than that in the crustaceans captured in the wild [2]. Similar results were obtained in a study of the effect of heavy metal ions on the amylolytic activity in chironomid larvae Chironomus plumosus (L.) [1]; this may be due to the greater resistance of individuals from natural populations to unfavorable environmen tal factors. Glycosidases from the tissues of juvenile roach were more sensitive to Roundup than zooplankton glycosi dases [2, 5]. Exposure to low concentrations of Roundup (0.1 mg/L) resulted in an increase in amy lolytic activity by 13–17% of the control in zooplank ton and juvenile roach, while exposure to higher con centrations (1–10 mg/L) did not affect the enzyme activity in either group of animals. Different changes in the activity of enzymes that hydrolyze polysaccha rides were detected in different species exposed to Roundup at concentrations of 25 and 50 mg/L: namely, the amylolytic activity of total crustacean zooplankton samples did not change; that in daphnia increased by 24 and 30%, respectively; and that in juvenile roach decreased by 11 and 24% from the con trol, respectively. The present study is the first to address changes in amylolytic activity, as well as those of maltase and sucrase activity, occurring in the tissues of juvenile fish and invertebrates from different ecological groups as a result of in vitro exposure to Roundup in a wide range INLAND WATER BIOLOGY
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of sublethal concentrations (0.1–50 µg/L). The lowest concentration tested is 10 times lower and the highest one is 50 times higher than the maximum permissible concentration. These concentrations can actually be observed in natural waters, since the glyphosate con centration in reservoirs with continuous vegetation amounted to 3.7 mg/L [17], this being equivalent to 9 mg/L Roundup. Enzyme activity in wholebody homogenates is a general characteristic of the activity of both digestive enzymes and the numerous lysosomal hydrolases from different organs and tissues. The effects of Roundup on glycosidase activity in wholebody homogenates of animals studied included both activation and inhibi tion. Roundup at a concentration of 0.1–50 µg/L had a weaker effect on amylolytic activity in the tissues of invertebrates than on that in juvenile fish. Roundup pretreatment resulted in an increase in maltase and sucrase activity in most invertebrates and juvenile fish studied. The concentrationdependent inhibitory effect was most clearly apparent in the body of actual prey, with amylolytic activity reduced by 19–32%; this was prob ably due to the activation of the numerous lysosomal glycosidases of the prey in the acidic medium of the predator stomach. Maltase activity in the tissues of real prey was reduced by 10–41% from that of the con trol and that of sucrase was reduced by 57–77% from the control; however, the effect did not depend on the concentration of the herbicide, this being indicative of the absence of a direct effect of Roundup on the active site of these enzymes. Similarly, the effects of chloro phos and nitrosoguanidine on the digestive glycosi dases of roach fingerlings did not depend on the con centration [3, 4]. The opposite effect of Roundup on glycosidases in the tissues of potential and actual prey is due to the differences in the functional activity of hydrolases at different pH values (neutral in the former case and acidic in the latter case). Because the prey enzymes may participate in selfdigestion processes after being released into the digestive tract of the pred ator [8], Roundup probably has a modifying effect on both the enzyme activity in the prey tissues and the degree of the exoenzyme contribution to the digestion in the predator. The mechanism of the toxic effect of Roundup and its components on animal glycosidases is still unclear; however, studying the physiological and biochemical parameters expands the understanding of the toxicity of the herbicide and makes it possible to predict the consequences of Roundup exposure for aquatic organisms. CONCLUSIONS A study of the effect of Roundup in a wide range of sublethal concentrations (0.1–50 µg/L) on glycosi dase (maltase, sucrase and amylolytic activity) in wholebody homogenates of invertebrates and juvenile INLAND WATER BIOLOGY
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fish showed that invertebrate enzymes are less sensitive to the herbicide than the enzymes of juvenile fish. The changes in glycosidase activity caused by Roundup exposure in the tissues of the animals investigated were of opposing characters; however, no correlation with the ecology of the species was identified. A greater inhibitory effect on glycosidase activity in tissues of actual prey than in potential prey is indicative of the inhibitory effect Roundup on the hydrolysis of carbo hydrates during actual gastric digestion and the reduc tion of the potential contribution of prey enzymes to digestion in the predator. Furthermore, this is indica tive of the possible dependence of the effect of Roundup on pH, which requires further experimental studies. REFERENCES 1. Golovanova, I.L., Kuz’mina, V.V., Urvantseva, G.A., et al., Effect of copper and zinc on the activity of carbo hydrase in young carp and chironomid larvae, in Sovre mennye problemy vodnoi toksikologii: Mater. Vseros. Konf. Borok (Modern Problems of Aquatic Toxicology: Proc. AllRussia Conf. Borok), Yaroslavl, 2002, p. 34. 2. Golovanova, I.L. and Papchenkova, G.A., The influ ence of the roundup herbicide on the activity of carbo hydrases in crustacean zooplankton and juvenile roach, Toksikol. Vestn., 2009, no. 4, pp. 32–35. 3. Golovanova, I.L. and Talikina, M.G., On the impact of low concentrations of chlorophos in the period of early ontogenesis on digestive carbohydrases of underyearlings of roach rutilus rutilus, J. Ichthyol., 2006, vol. 46, no. 5, pp. 404–408. DOI: 10.1134/S0032945206050079. 4. Golovanova, I.L., Talikina, M.G., Filippov, A.A., et al., Effect of ultralow concentrations of NmethylN' nitroNnitrosoguanidine upon early development in roach (Rutilis rutilus): intestine carbohydrase activities and kinetic characteristics of carbohydrate hydrolysis in the intestine of underyearlings, J. Ichthyol., 2008, vol. 48, no. 3, pp. 268–274. DOI: 10.1134/S0032945208030090. 5. Golovanova, I.L., Filippov, A.A., and Aminov, A.I., Effect of the roundup herbicide in vitro on the activity of carbohydrases in young fish, Toksikol. Vestn., 2011, no. 5, pp. 31–35. 6. Zhidenko, A.A. and Bibchuk, E.V., Changes in bio chemical parameters in the carp liver under exposure to the roundup herbicide, in Sovremennye problemy teoret icheskoi i prakticheskoi ikhtiologii: Tez. II mezhdunar. nauch.prakt. konf. (Modern Problems of Theoretical and Practical Ichthyology: Abstr. II Int. Sci.Pract. Conf.), Sevastopol, 2009, p. 50. 7. Zhidenko, A.A., Bibchuk, E.V., Krivopishina, V.V., and Barbukho, E.V., Changes in energy indices of the carp liver under herbicide load of different intensity, in Eko logicheskie problemy presnovodnykh rybokhozyaistven nykh vodoemov Rossii: Mater. Vseros. nauch. konf. (Environmental Problems of Freshwater Fishery Water Bodies in Russia: Proc. AllRussia Sci. Conf.), St. Petersburg, 2011, p. 167. 8. Kuz’mina, V.V., The contribution of induced autolysis in digestion in secondary consumers as exemplified by
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Translated by S. Semenova
INLAND WATER BIOLOGY
Vol. 6
No. 4
2013
