Cold hardiness,supercooling ability and causes of low-temperature mortality in the soft tick,Argas reflexus,and the hard tick,Ixodes ricinus (Acari: Ixodoidea) from Central Europe

Seasonal supercooling points (SCPs=temperature of crystallization) and cold hardiness were investigated in the indigenous hard tick, I. ricinus, and in A. reflexus, a soft tick introduced to Central Europe from the South. Both species proved to be freeze-susceptible as well as highly susceptible to inoculative freezing. None of the postembryonic developmental stages of either species showed any distinct seasonal pattern of SCP. Unexpectedly, the introduced A. reflexus exhibited a distinctly higher degree of cold hardiness in terms of lower lethal temperature (LT(50): 24h exposure) as well as lethal time (T(50): time of survival at -10.1 degrees C) than I. ricinus. Engorged I. ricinus larvae as well as engorged summer acclimatized A. reflexus larvae showed some mortality at temperatures well above the SCP. This mortality was generally expressed as a failure of the following stage to eclose properly. A 10-day cold acclimation at +3 degrees C eliminated that kind of mortality in summer acclimatized A. reflexus larvae, but not in I. ricinus larvae. It was frequently observed that freezing of ticks resulted-possibly via leakage from the midgut-in a subsequent reddish brown discoloration of the ticks after thawing. Taking into account that discoloration was an indication of previous freezing, it was concluded, that after long-term exposure (for >/=30 days) at -10.1 degrees C, a temperature well above the SCP, some tick mortality could be observed that was not caused by previous freezing. Weighing experiments clearly demonstrated, that the level of dehydration was not critical for survival of A. reflexus during long-term cold exposure, even at low RH. This indicates, that cold-related factors other than freezing and dehydration were detrimental to this species.     

The Siberian timberman Acanthocinus aedilis: a freeze-tolerant beetle with low supercooling points

Larvae of the Siberian timberman beetle Acanthocinus aedilis display a number of unique features, which may have important implications for the field of cold hardiness in general. Their supercooling points are scattered over a wide temperature range, and some individuals have supercooling points in the low range of other longhorn beetles. However, they differ from other longhorn beetles in being tolerant to freezing, and in the frozen state they tolerate cooling to below −37°C. In this respect they also differ from the European timberman beetles, which have moderate supercooling capacity and die if they freeze. The combination of freezing tolerance and low supercooling points is unusual and shows that freezing at a high subzero temperature is not an absolute requirement for freezing tolerance. Like other longhorn beetles, but in contrast to other freeze-tolerant insects, the larvae of the Siberian timberman have a low cuticular water permeability and can thus stay supercooled for long periods without a great water loss. This suggests that a major function of the extracellular ice nucleators of some freeze-tolerant insects may be to prevent intolerable water loss in insects with high cuticular water permeability, rather than to create a protective extracellular freezing as has generally been assumed. The freezing tolerance of the Siberian timberman larvae is likely to be an adaptation to the extreme winter cold of Siberia.     

Effects of diapause and cold-acclimation on the avoidance of freezing injury in fat body tissue of the rice stem borer, Chilo suppressalis Walker

Overwintering freeze-tolerant larvae of Chilo suppressalis can survive at -25 degrees C, but non-diapausing larvae cannot. We reported earlier that to prevent intracellular freezing, which causes death in overwintering larvae of the Saigoku ecotype distributed in southwestern Japan, water leaves and glycerol enters fat body cells through water channels during freezing. However, it is still unclear how diapause and low-temperature exposure are related to the acquisition of freeze tolerance. We compared the extent of tissue damage, accumulation of glycerol, and transport of glycerol and water in fat body tissues between cold-acclimated and non-acclimated non-diapausing and diapausing larvae. The tissue from cold-acclimated diapausing larvae could survive only when frozen in Grace's insect medium with 0.25 M glycerol at -20 degrees C. The protection provided by glycerol was offset by mercuric chloride, which is a water-channel inhibitor. Fat body tissue isolated from non-acclimated diapausing larvae was injured by freezing even though glycerol was added to the medium, but the level of freezing injury was significantly lower than in non-diapausing larvae. Radiotracer assays in cold-acclimated diapausing larvae showed that during freezing, water left the cells into the medium and glycerol entered the cells from the medium at the same time. Therefore, in Saigoku ecotype larvae of the rice stem borer, both diapause and cold-acclimation are essential to accumulate glycerol and activate aquaporin for the avoidance of freezing injury.     

The Effect of Household Smoking on the Infestiveness of Di Ph Ylloboth Riu M Latum from Fish to Man

The effect of household smoking, as carried out with three commercially available smoke boxes, on the infestiveness of Diphyllo-bothrium latum was investigated. The temperature changes in fish during the smoking were measured and the effect on tapeworm larvae estimated on the basis of a lethal temperature/time exposure of 56°C/5 min. For all the boxes smoking treatments sufficient to destroy the tapeworm larvae are suggested. The obtained results confirm the conclusion, made in previous studies on other heat preparation methods of fish, that well cooked fish in which all the parts have been subjected to a 56°C/5 min. exposure is safe with regard to the risk of Diphyllobothrium latum infestation.     

Energy utilization during swimming and cost of locomotion in larval and juvenile fish

Oxygen consumption and ammonia excretion rates increased in an accelerated manner in larvae and juveniles of whitefish (Coregonus sp.) as a function of swimming speed. The three-dimensional patterns of fish metabolic rates (expressed as energy consumed or nitrogen excreted) versus body weight and swimming speed show that the total standard metabolic rate (i. e. at extrapolated zero swimming speed) increased during early life of whitefish, followed by the expected decrease. This phenomenon might be due to the profound changes in oxidative and glycolytic enzyme activities during fish “metamorphosis”. Standard metabolic rate of ammonia excretion, as a principal product of protein catabolism in fish, decreased by one order of magnitude in early coregonid ontogenesis. This means that protein utilization as an energy source decreases as far as standard metabolism is concerned, but increases with swimming speed. This trend is opposite that in adult fish, where protein utilization in the overall energy supply is diminished at increasing swimming speed. The cost of locomotion offish larvae and juveniles demonstrates that the energy expenditure increases logarithmically with decreasing fish size but at an accelerated rate as compared to adult fish. This contradicts earlier estimates of lower cost of swimming in fish larvae than cost of paddle-propulsion swimming in small invertebrates or cost of flying in insects.     

False codling moth Thaumatotibia leucotreta (Lepidoptera,Tortricidae) larvae are chill‐susceptible

Abstract This study reports on the low temperature tolerance and cold hardiness of larvae of false codling moth, Thaumatotibia leucotreta. We found that larvae have mean critical thermal minima (lower limits of activity) of 6.7°C which was influenced by feeding status. The effects of low temperature exposure and duration of exposure on larval survival were assessed and showed that the temperature at which 50% of the population survives is ?11.5 ± 0.3°C after 2 h exposure. The supercooling point (SCP, i.e., freezing temperature) was investigated using a range of cooling rates and under different conditions (feeding and hydration status) and using inoculative freezing treatments (in contact with water or orange juice). The SCP decreased significantly from ?15.6°C to ?17.4°C after larvae were fasted for 24 h. Twenty‐four hour treatments at either high or low relative humidity (95.9% or 2.4%) also significantly decreased SCP to ?17.2°C and ?18.2°C respectively. Inoculative freezing (by water contact) raised SCP from ?15.6°C to ?6.8°C which could have important implications for post‐harvest sterilization. Cooling rates did not affect SCP which suggests that there is limited phenotypic plasticity of SCP during the larval life‐stage, at least over the short time‐scales investigated here. In conclusion, larvae of T. leucotreta are chill‐susceptible and die upon freezing. These results are important in understanding this pest's response to temperature variation, understanding pest risk status and improving post‐harvest sterilization efficacy.     

Synchrotron X-Ray Visualisation of Ice Formation in Insects during Lethal and Non-Lethal Freezing

Although the biochemical correlates of freeze tolerance in insects are becoming well-known, the process of ice formation in vivo is subject to speculation. We used synchrotron x-rays to directly visualise real-time ice formation at 3.3 Hz in intact insects. We observed freezing in diapausing 3^rd instar larvae of Chymomyza amoena (Diptera: Drosophilidae), which survive freezing if it occurs above −14°C, and non-diapausing 3^rd instar larvae of C. amoena and Drosophila melanogaster (Diptera: Drosophilidae), neither of which survive freezing. Freezing was readily observed in all larvae, and on one occasion the gut was seen to freeze separately from the haemocoel. There were no apparent qualitative differences in ice formation between freeze tolerant and non-freeze tolerant larvae. The time to complete freezing was positively related to temperature of nucleation (supercooling point, SCP), and SCP declined with decreasing body size, although this relationship was less strong in diapausing C. amoena. Nucleation generally occurred at a contact point with the thermocouple or chamber wall in non-diapausing larvae, but at random in diapausing larvae, suggesting that the latter have some control over ice nucleation. There were no apparent differences between freeze tolerant and non-freeze tolerant larvae in tracheal displacement or distension of the body during freezing, although there was markedly more distension in D. melanogaster than in C. amoena regardless of diapause state. We conclude that although control of ice nucleation appears to be important in freeze tolerant individuals, the physical ice formation process itself does not differ among larvae that can and cannot survive freezing. This suggests that a focus on cellular and biochemical mechanisms is appropriate and may reveal the primary adaptations allowing freeze tolerance in insects.     

Effects of summer frost exposures on the cold tolerance strategy of a sub-Antarctic beetle

The sub-Antarctic beetle Hydromedion sparsutum (Coleoptera, Perimylopidae) is common locally on the island of South Georgia where sub-zero temperatures can be experienced in any month of the year. Larvae were known to be weakly freeze tolerant in summer with a mean supercooling point (SCP) around -4 degrees C and a lower lethal temperature of -10 degrees C (15min exposure). This study investigated the effects of successive freezing exposures on the SCP and subsequent survival of summer acclimatised larvae. The mean SCP of field fresh larvae was -4.2+/-0.2 degrees C with a range from -1.0 to -6.1 degrees C. When larvae were cooled to -6.5 degrees C on 10 occasions at intervals of 30min and one and four days, survival was 44, 70 and 68%, respectively. The 'end of experiment' SCP of larvae surviving 10 exposures at -6.5 degrees C showed distinct changes and patterns from the original field population depending on the interval between exposure. In the 30min interval group, most larvae froze between -6 and -8 degrees C, a depression of up to 6 degrees C from the original sample; all larvae were dead when cooling was continued below the SCP to -12 degrees C. In the one and four day interval groups, most larvae froze above -6 degrees C, showing no change as a result of the 10 exposures at -6.5 degrees C. As with the 30min interval group, some larvae froze below -6 degrees C, but with a wider range, and again, all were dead when cooled to -12 degrees C. However, in the one and four day interval groups, some larvae remained unfrozen when cooled to -12 degrees C, a depression of their individual SCP of at least 6 degrees C, and were alive 24h after cooling. In a further experiment, larvae were cooled to their individual SCP temperature at daily intervals on 10 occasions to ensure that every larva froze every day. Most larvae which showed a depression of their SCP of 2-4 degrees C from their day one value became moribund or died after six or seven freezing events. Survival was highest in larvae with SCPs of -2 to -3 degrees C on day one and which froze at this level on all 10 occasions. The results indicate that in larvae in which the SCP is lowered following sub-zero exposure, the depression of the SCP is greatest in individuals that do not actually freeze. Further, the data suggest that after successive frost exposures in early winter the larval population may become segregated into two sub-populations with different overwintering strategies. One group consists of larvae that freeze consistently in the temperature range from -1 to -3 degrees C and can survive multiple freeze-thaw cycles. A second group with lower initial SCPs (around -6 degrees C), or which fall to this level or lower (down to -12 degrees C) after freezing on one or more occasions, are less likely to freeze through extended supercooling, but more likely to die if freezing occurs.     

The potential for cryopreserving larvae of the sea urchin, Evechinus chloroticus

Larvae of the sea urchin, Evechinus chloroticus, at varying stages of development, were assessed for their potential to survive cryopreservation. Ethylene glycol (EG) and dimethyl sulphoxide (Me2SO), at concentrations of 1-2 M, were evaluated as cryoprotectants (CPAs) in freezing regimes initially based on methods established for freezing larvae of other sea urchin species. Subsequent work varied cooling rate, holding temperature, holding time, and plunge temperature. Ethylene glycol was less toxic to larvae than Me2SO. However, no larvae survived freezing and thawing in EG. Larvae frozen in Me2SO at the gastrula stage and 4-armed pluteus stage regained motility post-thawing. The most successful freezing regime cooled straws containing larvae in 1.5 M Me2SO from 0 to -35 degrees C at 2.5 degrees C min(-1), held at -35 degrees C for 5 min, then plunged straws into liquid nitrogen. Motility was high 2-4 h post-thawing using this regime but decreased markedly within 24 h. Some 4-armed pluteus larvae that survived beyond this time developed through to metamorphosis and settled. Different Me2SO concentrations and supplementary trehalose did not improve long-term survival. Large variation in post-thaw survival was observed among batches of larvae produced from different females.     

Effects of temperature on larval fish swimming performance: the importance of physics to physiology

Temperature influences both the physiology offish larvae and the physics of the flow conditions under which they swim. For small larvae in low Reynolds number (Re) hydrodynamic environments dominated by frictional drag, temperature‐induced changes in the physics of water flow have the greatest effect on swimming performance. For larger larvae, in higher Re environments, temperature‐induced changes in physiology become more important as larvae swim faster and changes in swimming patterns and mechanics occur. Physiological rates at different temperatures have been quantified using Q₁₀s with the assumption that temperature only affected physiological variables. Consequently, Q₁₀s that did not consider temperature‐induced changes in viscosity overestimated the effect of temperature on physiology by 58% and 56% in cold‐water herring and cod larvae respectively. In contrast, in warm‐water Danube bleak larvae, Q₁₀s overestimated temperature‐induced effects on physiology by only 5–7%. This may be because in warm water, temperature‐induced changes affect viscosity to a smaller degree than in cold water. Temperature also affects muscle contractility and efficiency and at high swimming velocities, efficiency decreases more rapidly in cold‐exposed than in warm‐exposed muscle fibres. Further experiments are needed to determine whether temperature acts differently on swimming metabolism in different thermal environments. While hydrodynamic factors appear to be very important to larval fish swimming performance in cold water, they appear to lose importance in warm water where temperature effects on physiology dominate. This may suggest that major differences exist among locomotory capacities of larval fish that inhabit cold, temperate waters compared to those that live in warm tropical waters. It is possible that fish larvae may have developed strategies that affect dispersal and recruitment in different aquatic habitats in order to cope not only with temperature‐induced physiological challenges, but physical challenges as well.     

Bioenergetic model of planktivorous fish feeding, growth and metabolism: theoretical optimum swimming speed of fish larvae

The feeding activity of an individual fish larva is described by an equation which includes parameters for the area successfully searched, probability of food capture multiplied by the cross-sectional perceptive visual field, larval swimming speed and the time required to consume a unit of food energy. The proportion of ingested food energy used for metabolism increases exponentially with increasing swimming speed. The model predicts that food consumption rate increases asymptotically whereas metabolic rate increases exponentially. This results in a predicted growth rate curve that reaches a maximum at a certain swimming speed and decreases at both higher and lower speeds. The model can be used to predict the influence of type of prey, prey density, water temperature etc. on larval growth. An expression describing how many hours per day fish larvae must forage in order to grow at a certain daily body weight gain allows the limits of environmental conditions for positive, zero and negative growth rate to be set. Results of simulations demonstrated that the optimum swimming speed for maximum growth of coregonid larvae increased with an increase in food density, decrease in water temperature or decrease of prey vulnerability. At optimum ‘theoretical’ swimming speed an increase in water temperature from 5 to 17° C required the food density to be increased from 20 to 80 copepods l^?1 in order to maintain a daily growth increment of 2%. The minimum Artemia density required for maintenance metabolism increased from 10 to 30 items 1¹ over the same temperature increase from 5 to 17° C, and food densities required for 8% growth rates were 26 and 56 Artemia nauplii l^?1 at 5 and 17° C, respectively. Contrary to previous findings, results of the present study suggest that metabolic rates of actively feeding fish larvae may be from 5 to 50 times the standard metabolic rate: earlier studies suggested that a factor of 2–3 may be generally applicable.     

Acquisition of freezing tolerance in early autumn and seasonal changes in gall water content influence inoculative freezing of gall fly larvae,Eurosta solidaginis (Diptera,Tephritidae)

We examined seasonal changes in freeze tolerance and the susceptibility of larvae of the gall fly, Eurosta solidaginis to inoculative freezing within the goldenrod gall (Solidago sp.). In late September, when the water content of the galls was high (approximately 55%), more than half of the larvae froze within their galls when held at -2.5 degrees C for 24 h, and nearly all larvae froze at -4 or -6 degrees C. At this time, most larvae survived freezing at > or = -4 degrees C. By October plants had senesced, and their water content had decreased to 33%. Correspondingly, the number of larvae that froze by inoculation at -4 and -6 degrees C also decreased, however the proportion of larvae that survived freezing increased markedly. Gall water content reached its lowest value (10%) in November, when few larvae froze during exposure to subzero temperatures > or = -6 degrees C. In winter, rain and melting snow transiently increased gall water content to values as high as 64% causing many larvae to freeze when exposed to temperatures as high as -4 degrees C. However, in the absence of precipitation, gall tissues dried and, as before, larvae were not likely to freeze by inoculation. Consequently, in nature larvae freeze earlier in the autumn and/or at higher temperatures than would be predicted based on the temperature of crystallization (T(c)) of isolated larvae. However, even in early September when environmental temperatures are relatively high, larvae exhibited limited levels of freezing tolerance sufficient to protect them if they did freeze.     

Oxygen uptake during early life in the fresh water fish, Esomus danricus (Ham) (Pisces, Cypriniformes)

O2 uptake in Esomus danricus has been determined in relation to body weight, length and thickness of the water-blood diffusion barrier at 27-28 degrees C temperature. Total O2 consumption in larvae was 1311 ml/kg/h but decreased significantly in juvenile fishes (720 ml/kg/h). The increase in the thickness of water-blood diffusion barrier at the secondary gill lamellae of the fish was found to be an important factor for the decrease in VO2. Logarithmic analyses of data for O2 uptake in relation to body weight gave a slope of 0.8865 for larvae and 0.5053 for juveniles. The exponent values of O2 uptake against diffusion barrier for larvae and juveniles were 1.7383 and 2.0956, respectively. The results obtained indicated that fish have an extra device which helps in extracting about 24% of the total VO2 required for the fulfilment of the metabolic oxygen demand of the body.     

The Effect of Low Temperatures on the Motility of Diphyllobothrium Latum Plerocercoids

The effect of low temperature exposure on the motility of Diphyl-lobothrium latum plerocercoids was studied, with the particular aim of finding the exposure that immobilizes all the larvae in fish freezing. Both isolated larvae immersed in normal horse serum and larvae enclosed in pieces of muscle tissue of the size of 1 cm3 were tested. The pieces of tissue containing a larva were placed in the middle of a plastic beaker filled with densely packed minced fish flesh. In the central part of this phantom, where the plerocercoids were situated, the temperature decline was considered to take place in the same way as in the interior of a whole fish. A total of 200 isolated larvae were tested, and a temperature of −14° G was found to have a fully immobilizing effect on them. The number of plerocercoids frozen enclosed in muscular tissue was 453, and −10° G was found to immobilize them. The observed difference seems to be mainly due to the cryoprotective properties of serum.     

Effects of food density and temperature on feeding and growth of young inland silversides (Menidia beryllina)

Food consumption and growth rates of 7–28-day-old Menidia beryllina were measured in response to natural ranges of temperature and prey availability. Feeding level, temperature and age all had significant effects on growth rate, although the effect of feeding level explained most of the variance. Feeding level also had a significant effect on gross growth efficiency, but temperature and age did not. Absolute growth rates (mg per day) increased dramatically with temperature, feeding level, and age. Variability in growth was greatest for fish feeding at the lowest feeding level. For a given fish weight, temperature had a positive effect on consumption rate, and maximum consumption ( C _max) of any treatment combination reached 75% body weight per day. Maximum growth rate was estimated at 24.6% body weight per day, and gross growth efficiency reached an estimated maximum of 0.375 at an ingestion rate of 25% body weight per day. Starved larvae lost on average 5.4% body weight per day and larvae required 6.4% body weight food consumption per day for maintenance. Multiple regressions of feeding level, temperature, and age/size on instantaneous growth rates indicated that increases in temperature increased maintenance requirements and required that fish consume a greater proportion of C _max to attain maximum growth. Growth rates decreased with increases in temperature for fish eating a specific weight of food.     

Consumption and gut evacuation rate of laboratory‐reared spotted seatrout (Sciaenidae) larvae and juveniles

The temperature and mass dependence of maximum consumption rate was measured for larval and early juvenile spotted seatrout Cynoscion nebulosus . Maximum consumption ( C _MAX) estimates were obtained from feeding and gut evacuation experiments on larvae (3·8–19 mm standard length, L _S) at three temperatures (24, 28 and 32° C), and maximum consumption experiments on juveniles at three temperatures (20, 26 and 32° C). Feeding levels were determined for larvae fed live prey ( Brachionus plicatilis and Artemia salina ) ad libitum . The midgut and total evacuation times were estimated for fish feeding continuously and discontinuously using alternate meals of tagged and untagged live prey. Temperature and fish size had significant effects on gut evacuation and consumption. The gut evacuation time increased with increasing fish size, and decreased with increasing temperatures. Mass‐specific midgut contents increased for small larvae <0·156 mg dry mass ( M _D)( c . 4 mm L _S), and decreased for larger larvae and juveniles. Maximum consumption was modelled by fitting a polynomial function to a reduced dataset of individuals feeding at high levels. The C _MAX model predicted an initial increase in specific feeding rate from 70 to 155% M _D day⁻¹ for small larvae, before declining for larger larvae and juveniles.     

Food selection and growth of young snakehead Channa striatus

Food selection and growth of young snakehead Channa striatus were studied in the laboratory and in a field trial. In the laboratory, first-feeding snakehead larvae of 6–7 mm total length (TL) with a mouth opening of 0.55 mm selected for Artemia nauplii, and against formulated feed. Fish began feeding on formulated feed at 12mm TL when their mouth width reached 1.0mm. In both laboratory and field trials, snakehead diets changed as fish size increased. For fish 15–20 mm TL, cladocerans and copepods were 96.5% of their diet. With fish 30–40 mm TL, zooplankton consumption was greatly reduced while benthic organism consumption increased. Fish 45–50 mm TL fed exclusively on benthic invertebrates. Diet shift from zoo-plankton to benthic invertebrates was not due to reduced zoo-plankton availability, but was instead related to changes in gill raker structure. Low density of benthic invertebrates in the field trial caused reduced fish growth rates when fish switched diets from zooplankton to benthos. Our results indicate that snake-head can take Artemia nauplii as a larval starter food, then accept formulated feed at ≥12 mm TL. Zooplankton can serve as food for snakehead < 40 mm, but formulated feed should be provided for larger fish which are unable to catch zoo-plankton.     

Water temperature determines strength of top-down control in a stream food web

1. We examined effects of water temperature on the community structure of a three trophic level food chain (predatory fish, herbivorous caddisfly larvae and periphyton) in boreal streams. We used laboratory experiments to examine (i) the effects of water temperature on feeding activities of fish and caddisfly larvae and on periphyton productivity, to evaluate the thermal effects on each trophic level (species‐level experiment), and (ii) the effects of water temperature on predation pressure of fish on abundance of the lower trophic levels, to evaluate how temperature affects top‐down control by fish (community‐level experiment). 2. In the species‐level experiment, feeding activity of fish was high at 12 °C, which coincides with the mean summer temperature in forested streams of Hokkaido, Japan, but was depressed at 3 °C, which coincides with the mean winter temperature, and also above 18 °C, which coincides with the near maximum summer temperatures. Periphyton productivity increased over the range of water temperatures. 3. In the community‐level experiments, a top‐down effect of fish on the abundance of caddisfly larvae and periphyton was clear at 12 °C. This effect was not observed at 3 and 21 °C because of low predation pressure of fish at these temperatures. 4. These experiments revealed that trophic cascading effects may vary with temperature even in the presence of abundant predators. Physiological depression of predators because of thermal stress can alter top‐down control and lead to changes in community structure. 5. We suggest that thermal habitat alteration can change food web structure via combinations of direct and indirect trophic interactions.     

Fly Larvae in Wastewater Discharges: A Potential Source of Food and Organochlorine Contaminants for some Southern California Fishes

A widely recognized pathway for uptake and accumulation of organic contaminants by demersal fishes is through consumption of infaunal organisms (i.e., those having direct contact with contaminated sediments). Recent studies indicate that demersal fishes near a large sewage outfall in southern California consume larvae of a terrestrial fly, Clogmia albipunctata, which are entrained with the treated effluent and discharged to the ocean. Fly larvae consumption represents a possible method for accumulation of organic contaminants that is unique to marine fishes with non-selective feeding habits that occur near the ocean outfall. An estimated 37–138kg (wet weight) of fly larvae are discharged daily during summer to the ocean via the Orange County Sanitation Districts (OCSD) wastewater outfall; winter discharges are expected to be lower due to lower production and temperature limitations of the fly life cycle. Fly larvae were present in the guts of ten demersal and pelagic fish species collected near the outfall between July 1993 and August 1998. The highest frequencies of fly larvae (up to 100%) occurred in the guts of white croaker, Genyonemus lineatus, and chub mackerel, Scomber japonicus. Because infauna are still abundant and diverse near the outfall, fly larvae consumption appears to represent a feeding preference instead of a response to lower prey abundances. Fly larvae tissues collected at the wastewater treatment plant contained sub-part-per-million levels of organochlorine compounds, including PCB and DDT. However, lipid-normalized organochlorine concentrations in fly larvae tissues were not appreciably higher than levels in infauna or zooplankton tissues. Although the study design provided only limited sample sizes for some sample types, this initial study suggests that fish consumption of fly larvae represents a mechanism for direct transfer of contaminants and may contribute to uptake and accumulation of lypophyllic organochlorines in fish near the outfall.     

Influence of storage temperature and freezing time on histamine level in the European anchovy Engraulis encrasicholus (L., 1758): A study by capillary electrophoresis

Histamine content in fish may increase by decarboxylation of free histidine to values that can be toxic, if storage conditions are not well controlled. We have studied the influence of storage temperature and time of freezing on histamine formation in the anchovy, Engraulis encrasicholus (L., 1758), for which little information is available. Analysis, carried out by capillary zone electrophoresis (CZE) without sample pre-treatment, was very simple, fast and reproducible. Results indicate that temperatures above 20 degrees C notably increase histamine production, whereas freezing can clearly prevent or slow down the process.