Scope for Growth of the Estuarine Mysid Neomysis Integer (Peracarida: Mysidacea): Effects of the Organophosphate Pesticide Chlorpyrifos

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Scope for Growth of the Estuarine Mysid Neomysis Integer (Peracarida: Mysidacea): Effects of the Organophosphate Pesticide Chlorpyrifos MARINE ECOLOGY PROGRESS SERIES Published December 30 Mar Ecol Prog Ser / Scope for growth of the estuarine mysid Neomysis integer (Peracarida: Mysidacea): effects of the organophosphate pesticide chlorpyrifos 'Plymouth Environmental Research Centre (Department of Biological Sciences), University of Plymouth. Drake Circus, Plymouth, Devon PL4 8AA, United Kingdom 'Centre for Coastal and Marine Sciences, Plymouth Marine Laboratory, Prospect Place. West Hoe. Plymouth, Devon PLl 3DH, United Kingdom ABSTRACT: Myslds (Peracanda: Mysldacea) are standard test organisnls used routinely in acute tests to evaluate the comparative toxicity of chemicals to aquatic organisms. The criticism of such lethal mea- surements for ecosystem monitoring has resulted in a clear need for sub-lethal tests that enable a more comprehensive understanding of the potential lmpacts of toxicants on natural populations Neomysis integer, the dominant mysid of the upper reaches of European estuaries, has relatively low 96 h LCS0 values for pesticides and trace metals, and is under investigation as a suitable species for monitonng the pollution status of European estuaries In thls paper, the effects of environmentally realistic con- centratlons of the organophosphate pesticide chlorpyrifos on some basic physiological processes of N. Integer are evaluated as potential end points for Indicators of environn~entalpollution Oxygen con- sumption, egestion rate (used as a measure of fcc:llng rate) and scope lor growth (an integrated mea- sure of the overall physiological status of an organism) werc. significantly affected by chlorpyrifos expo- sure. In contrast, absorption efficiency was unaffected by pesticide exposure. Chlorpyrifos effects were most pronounced following short (i.e. 48 h) compared with longer exposure periods (e.g 168 h). Increased oxygen consumption was the most sensitive toxicity test end point for detecting chlorpyrifos exposure, however, this response may not apply to all toxlcants. Reduced scope for growth and eges- tion rate are considered more representatwe of the effects of chlorpyrifos on natural mysid populations, but were only affected significantly at near-lethal exposure concentrations. All 3 responses were affected at concentrations lower than those currently acceptable by European Community legislation for potable waters KEY WORDS: Neomysjs integer - Sub-lethal testing . Chlorpyrifos . Organophosphate pesticide INTRODUCTION environmental impact Several physiological re- sponses have been used to quantify the sub-lethal Acute, lethal toxicity tests provide useful data on the effects of contaminants on crustaceans. These include comparative toxicity of chemical contaminants to respiration (e.g.Gaudy et al. 1991),feeding (e.g.Crane aquatic b~otabut are of limited value for environmen- & Maltby 1991),excretion (e.g.McKenney & Matthews tal management, part~cularlywhen the LCS0values are 1990) and reproduction (e.g. McKenney & Celestial higher than the toxicant concentrations measured in 1996). In addition, 2 or more of these individual the natural environment. Consequently, there is an responses may be combined into a single, integrated increasing need to develop tests which measure sub- bioassay such as 'scope for growth' (SFG) which pro- lethal responses to toxicants at environmentally realis- vides a rapid, instantaneous measurement of the tic concentrations and which may be used to predict energy status of an organism (Widdows 1993).To date, there have been few attempts to compare different physiological responses of the same specles to deter- O Inter-Research 1999 Resale of full art~clenot permitted 234 Mar Ecol Prog Ser 191: 233-241, 1999 mine which is the most sensitive, or most appropriate, Pesticide exposure regime. Mysids were exposed to for environmental monitoring (cf. Bamber & Depledge chlorpyrifos concentrations of 0.038, 0.056, 0.072 and 1997). In this paper, the effects of the pesticide 0,100 pg 1-I for 3 time periods (48, 96 and 168 h).Three chlorpyrifos (O,O,-diethyl 0-3,5,6-trichloro-2-pyridyl pesticide concentrations were predicted to be sub- phosphorothioate), a non-systemic organophosphoro- lethal, based on a 168 h LCS0 estimate of 0.084 pg thioate insecticide (Whitehead 1997), on the respira- chlorpyrifos I-' for adult Neomysis integer (Roast et al. tion, feeding, absorption efficiency and SFG of the 1999a);0.100 pg I-' chlorpynfos was i.ncluded to exam- estuarine mysld Neomysis integer (Leach) are com- ine pesticide effects at near lethal concentrations. Each pared. The potential of each parameter as a toxicity chlorpyrifos concentration disrupts mysid swimming test end point is assessed. Surprisingly, although SFG behaviour (Roast et al. 1999b).Due to the hydrophobic has been reported for other peracarid crustaceans (e.g. nature of chlorpyrifos [log K,, = 4.7 (Whitehead 1997)], the amphipod Gammarus pulex [Maltby et al. 1990]), Distol grade acetone was used as a carrier for pesticide it has not been evaluated previously for any mysid dosing (Roast et al. 1999a).An initial stock of 1 g chlor- species. pyrifos I-' acetone was diluted in series to produce a Mysids (Crustacea: Peracarida) are used routinely in final stock concentration of 1 mg 1-' acetone; dilution laboratory toxicity tests and the sub-tropical American water (salinity = 10 psu) was made by combining fil- species, Americamysis (=Mysidopsis)bahia, is a stan- tered seawater and double-distilled, de-ionised water. dard test organism adopted by the United States Envi- Toxicant stock (76, 112, 144 or 200 p1) was injected via ronmental Protection Agency (USEPA) for marine and a Gilson pipette into the vortex of 1 1 of dilution water estuarine studies (e.g. USEPA 1987). Recent work has stirred at high speed in a 2 1 conical flask. Equalising shown that A. bahia has limited tolerance to dilute volumes of acetone were injected into each vessel such salinity, casting doubt on its value as a test organism that each had the same concentration of acetone (thus for use in estuaries (Pillard et al. 1999). More impor- 124, 88, and 56 p1 acetone were injected into the 0.038, tantly, A. bahia is not representative of European estu- 0.056 and 0.072 pg chlorpyrifos 1-' solutions, such that aries and there is a clear need to identify an indigenous each exposure solution contained 100 p1 acetone I-'). estuarine species for toxicity testing in Europe (e.g. Following 5 min mixing, the conical flask was filled to NRA 1993). Neomysis integer, the dominant hyperben- an accurately pre-marked 2 l line to give the final con- thic mysid of the upper reaches of European estuaries centrations required. In addition to the 4 pesticide (Mees et al. 1995, Mees & Jones 1997), has relatively exposure solutions, a solvent control (100 p1 acetone low 96 h LCS0values for cadmium (3.9 pg 1-' [Wildgust 1-l) was used to assess any effects of acetone on mysid & Jones 19981) and the organophosphate pesticides physiology. As oxygen consumption by N. integer is chlorpyrifos and dimethoate (0.14 pg 1-' and 0.54 mg 1-' gender specific (Roast et al. 1999c), only males were respectively [Roast et al. 1999al).N. integer has been used in these experiment. For each exposure concen- proposed as a test species for monitoring European tration, 10 mysids of equal length (12 + 1 mm from the estuaries (Emson & Crane 1994, Roast et al. 1998a) and anterior margin of the rostrum to the tip of the telson) the present paper contributes further to this discussion. were placed together in 2 1tall-form glass beakers con- taining 1500 m1 of exposure wa.ter. Due to the unstable nature of chlorpyrifos (ca 55% loss within 24 h [Roast MATERIALS AND METHODS et al. 1999a]),exposure solutions were replaced every 24 h with a freshly made solution. At this tim.e, dead or Mysid collection and maintenance. Du.ring March to moribund mysids were removed from the test vessels May 1997, Neomysis integer were collected from the (mortality was concentration dependent, and up to ca southern side of Terras Bridge, East Looe River Estuary 60% had died after 7 d exposure to 0.1 pg chlorpyrifos (Cornwall, UK) (Roast et al. 1998b). Mysids were 1-'). Mysids were fed ad libitum on 148 h old Artemia returned to the laboratory in habitat water (salinity = sp. nauplii. 1 psu) and placed in a shallow 15 1 holding tank at a Measurement of oxygen consumption. Following salinity of 10 + 1 psu (made by combining filtered sea- exposure to chlorpyrifos for each time period (48, 96 water and double-distilled, de-ionlsed water) In a con- or 168 h), a single mysid was placed in a 25 m1 per- stant-temperature room (15 + 1°C). Water quality was spex respiration chamber designed for 'closed' oxy- maintained by an under-gravel filter. Lighting was gen measurement (Roast et al. 1999~).Oxygen con- provided at ambient laboratory levels by overhead flu- sumption was measured using Strathkelvin 1302 orescent lights; a time switch provided a 16 h light:8 h 'Clark-type' polarographic electrodes and Strath- dark photoperiod. Mysids were fed ad libitum with kelvin 781b oxygen meters, calibrated with oxygen- <48 h old Artemia sp. (Great Lakes, Utah) nauplii free nitrogen and air-saturated water, using values hatched in the laboratory from cysts. from dissolved oxygen tables (Green & Carritt 1967). Roast et al.. Pesticides and mysid physiology 235 Oxygen consumption measurements were performed weighed (*0.01 mg) to ascertain the ash content of the at 15 c 1°C (maintained by placing the respiration faeces. Absorption efficiency was calculated using the chamber in a temperature-controlled water bath) and equation: at 10 c 1 psu, using freshly made exposure solution Absorptionefficiency = (F-E)+[(l-E)xF] (1) of the same chlorpyrifos concentration to which the mysid had been exposed. Dissolved oxygen concen- where: F=ash-free dry weight:dry-weight ratio of food trations in each respiration chamber were measured source, and E= ash-free dry weight:dry-weight ratio of for 2 h prior to introduction of the mysids to confirm faeces (Conover 1966).
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