Relationship between stream temperature, thermal refugia and rainbow trout Oncorhynchus mykiss abundance in arid-land streams in the northwestern United States

Abstract – Warm stream temperatures may effectively limit the distribution and abundance of Pacific salmon Oncorhynchus spp. in streams. The role of cold thermal refugia created by upwelling groundwater in mediating this effect has been hypothesized but not quantitatively described. Between June 21 and September 15, 1994, rainbow trout O. mykiss abundance within 12 northeast Oregon (USA) stream reaches was inversely correlated with mean ambient maximum stream temperatures ( r =−0.7, P <0.05). Some rainbow trout used thermal refugia (1–10 m² surface area) that were on average 3–8°C colder than ambient stream temperatures. Within the warmest reaches, high ambient stream temperatures (>22°C) persisted from mid-June through August, and on average 10–40% of rainbow trout were observed within thermal refugia during periods of midday maximum stream temperatures. Frequency of cold-water patches within reaches was not significantly associated with rainbow trout density after accounting for the influence of ambient stream temperature ( P =0.06; extra sum of squares F -test). Given prolonged high ambient stream temperatures in some reaches, the thermal refugia available in the streams we examined may be too small and too infrequent to sustain high densities of rainbow trout. However, these refugia could allow some rainbow trout to persist, although at low densities, in warm stream reaches.     

Photobacterium damselae subsp. damselae, an emerging pathogen in Danish rainbow trout, Oncorhynchus mykiss (Walbaum), mariculture

A selection of 16 field isolates of Photobacterium damselae from marine rainbow trout farms in Denmark was subjected to phenotypic and genotypic characterization and pathogenicity to fish. All isolates belonged to the subspecies damselae , being positive for haemolysis, motility and urease. There were considerable differences in haemolytic properties, some isolates presenting a broad zone of haemolysis and others only a narrow zone. Pulsed-field gel electrophoresis revealed a high diversity indicating that P. damselae subsp. damselae is an opportunistic, not clonal pathogen in Danish marine rainbow trout. Virulence of the strains to rainbow trout was highly variable with LD₅₀ values ranging from 3.9 × 10³ to 1.5 × 10⁸ cfu at 20 °C. The virulence was significantly higher at 20 °C than at 13 °C. The strains with the strongest haemolytic properties were the most virulent suggesting a strong involvement of haemolysin in the pathogenesis. The pathological changes were consistent with a bacterial septicaemia and the haemorrhages were more pronounced than for most other bacterial infections.     

Differences in self-feeding activity between thermally selected and normal strains of rainbow trout Oncorhynchus mykiss at high temperatures

ABSTRACT:   Feeding activity was examined at high temperatures by using a demand feeder for thermally selected and normal rainbow trout Oncorhynchus mykiss strains. When the water temperature was raised in experiment 1 from 17.5 to 25.7°C at 0.3°C/day in 21 days, the average daily food consumption rate in the thermally selected strain rose to 7.1%, which was significantly higher than that of the normal strain (4.1%, P  < 0.05). The corresponding rate was also significantly higher in the thermally selected (0.8%) than in the normal strain (0.2%, P  < 0.05) of fish in experiment 2 where water temperature was raised to 24.1°C in 0.5°C/day increments. When water temperature was raised rapidly in experiment 3 from 16.7 to 21.7°C in one day and gradually to 24.4°C in 28 days at 0.1°C/day, the average daily food consumption rates were 1.0 and 0.1% for the thermally selected and normal strains, respectively, with significant difference ( P  < 0.01). These results suggest that the thermally selected strain has acquired thermal tolerance as a result of artificial selection.     

A bath challenge model for furunculosis in rainbow trout, Salmo gairdneri Richardson, and Atlantic salmon, Salmo salar L.

Abstract. A natural bath challenge method has been developed for furunculosis using Atlantic salmon, Salmo salar L., and rainbow trout, Salmo gairdneri Richardson. Fish were placed in an enclosed, continuously circulating tank system, supplemented with additional oxygen, the temperature raised gradually (overnight) to 15–16°C, a low dose of Aeromonas salmonicida (strain 184/186) introduced into the tank and the challenge maintained for 14 days. The challenge strain was characterized with respect to possible virulence factors and possessed an A-layer, ability to auto-agglutinate, haemagglutinate, adhere to Atlantic salmon cells and resist destruction by serum. No caseinase activity was detected. LD₅₀ for salmon using this method was 1.8×10³ cells per millilitre while trout had an LD₅₀ of 9.5×10⁴ cells per millilitre. Onset of the disease appeared to depend on fish size with larger trout (50 g) taking up to 10 days to show signs of the disease while mortalities in smaller trout (8.5g) commenced on day 1 post-challenge.     

Evidence of concealment behavior by adult rainbow trout and brook trout in winter

Abstract – There has been little investigation of the winter ecology of adult trout during winter, especially in regard to concealment behavior. We compared day vs night underwater counts of adult rainbow trout and brook trout from four streams. At water temperatures between 1°C and 9°C, daytime counts accounted for 44% and 16% of nighttime snorkeling counts for rainbow trout and brook trout adults, respectively. As winter progressed, nighttime counts declined more so for brook trout than rainbow trout, but the decline was not significant for either species. Nocturnalism of both species was higher in streams with colder water temperatures. We observed few fish within concealment structure; however, by electrofishing concealment habitat during the day, we captured 10 times more adult trout than we counted immediately beforehand by snorkeling. Adult trout were concealed in cobble-boulder substrate and woody debris during the day. ^Note     

The non-specific activation of rainbow trout, Salmo gairdneri Richardson, complement by Aeromonas salmonicida extracellular products and the correlation of complement activity with the inactivation of lethal toxicity products

Abstract The possible mechanism of inactivation of the toxicity of Aeromonas salmonicida extracellular products (ECP) by normal rainbow trout serum was investigated using juvenile rainbow trout. ECP was prepared from culture supernatant by an acetone precipitation method. The ECP was incubated with normal rainbow trout serum at 20°C for 2 h, and the interrelationship between ECP proteolytic activity and immune complex-initiating, haemolytic complement activity (CH₅₀) of normal serum against antibody-sensitized goldfish red blood cells was evaluated. When normal serum was incubated with increasing concentrations of ECP, the CH₅₀ activity of serum decreased. The CH₅₀ activity was completely abolished in serum treated with undiluted ECP. ECP treated with serum was administered to trout intraperitoneally to determine mortality. All the fish receiving untreated ECP (0.05 ml = 0.5 mg protein) alone died within 24 h. When ECP was treated with serum at 1:1 to 4:1 (serum: ECP) in volume a similar high mortality was produced. These inocula possessed high protease activity and no or low CH₅₀ activity. However, mortality decreased and finally no mortality was recorded as ECP was treated with large volumes of serum (9:1 to 19:1). These inocula had lower protease activity and considerably higher CH₅₀ activity. Fish receiving ECP treated with heat-inactivated serum at 19:1 showed 100% mortality. A serum: ECP inoculum derived from fish which had been administered lipopolysaccharide from Salmonella enteritidis and which possessed a low CH₅₀ activity also gave a high mortality when used at 19:1. These results suggest that rainbow trout complement is implicated in the inactivation of toxicity of A. salmonicida ECP.     

Distended, water-filled stomach in seawater farmed rainbow trout, Oncorhynchus mykiss (Walbaum), provoked experimentally by osmoregulatory stress

The present study evaluates if abdominal dis-tension caused by a water-filled stomach in seawater farmed rainbow trout, Oncorhynchus mykiss (Walbaum), and Atlantic salmon, Salmo salar L., could be provoked experimentally by live chilling in sea water (0.5 °C). Fifty rainbow trout and 50 salmon were visually classified, either as normal or suffering from the condition. Prior to chilling, no rainbow trout or salmon suffered from a water-filled stomach. During chilling, 25% of the rainbow trout and 2.5% of the salmon developed water-filled stomach. Affected rainbow trout had significantly higher blood plasma osmolality and significantly lower body weight than normal trout. The frequency of trout with a water-filled stomach increased significantly with increasing chilling time and increasing plasma osmolality. The regression coefficients revealed that the proportion of affected rainbow trout in-creased by 0.82% h⁻¹ of chilling and by 0.46% per unit increase in mosmol L^−1. The present study revealed that abdominal distension in seawater farmed rainbow trout was far more severe than in Atlantic salmon and could be provoked experimentally by osmoregulatory stress. This is consistent with the observation that water-filled stomach appear only occasionally in seawater farmed Atlantic salmon.     

Changes in enzymatic and structural properties of grass carp fast skeletal myosin induced by the laboratory-conditioned thermal acclimation and seasonal acclimatization

ABSTRACT:   Myosins were prepared from fast skeletal muscles of grass carp thermally acclimated to 10, 20 and 30°C in the laboratory as well as from those seasonally acclimatized and collected in January (winter) 2003 and May (spring), August (summer) and November (autumn) 2002. The maximal initial velocities ( V _max) of actin-activated Mg²⁺-ATPase activity for myosins from the 10°C-acclimated and winter grass carp were 1.7–1.8-fold as high as those from the 30°C-acclimated and summer fish. The inactivation rate constant ( K _D) of Ca²⁺-ATPase for myosin from the 10°C-acclimated grass carp was three to fourfold higher than those for myosins from the fish acclimated to 20°C and 30°C, whereas myosin from winter grass carp was about sevenfold as high as that for myosin from summer fish. Myosins from spring and autumn fish showed K _D values comparable to those of the fish acclimated to 30°C and 10°C, respectively. In differential scanning calorimetry analysis, the transition temperature ( T _m) was observed near 38°C and 45–46°C with most myosins. However, the lowest T _m at 32–33°C was given as one of the major endotherms in myosins from the 10°C-acclimated, autumn and winter fish. These responses of grass carp to changed environmental temperatures were almost similar to those for common carp reported previously.     

Effects of dietary protein and lipid level, and water temperature, on the post-feeding oxygen consumption of Atlantic cod and haddock

Tank respirometry was used to study the interactive effects of protein:lipid level (55%:11% vs. 42%:16%; both diets isoenergetic) and temperature (11, 6 and 2 °C) on the magnitude and duration of specific dynamic action (SDA) in juvenile Atlantic cod and haddock. The protein:lipid level did not affect any measured variable. However, numerous temperature and species effects were observed. For example, although the maximum post-feeding oxygen consumption (30–50% above routine metabolic rate; RMR) and SDA duration (∼55–85 h; SDA_DUR) were not affected by temperature, SDA_DUR g⁻¹ of food increased from 11 to 2 °C (from ∼3 to 12 h g food⁻¹). While absolute SDA (mg O₂) decreased by ∼60–65% in cod and ∼75% in haddock from 11 to 2 °C, due to a concomitant decrease in food consumption from ∼2.0% to 0.6% body mass, SDA comprised between 3.3% and 5.2% of the dietary energy content at all temperatures. Finally, RMR at 11 and 2 °C and SDA_DUR at 2 °C were 25–35% and 25% greater in cod, respectively, as compared with haddock. These results suggest that feeding reduced protein diets at low water temperatures is unlikely to improve the growth of these species.     

Susceptibility of selected freshwater fish species to a UK Lactococcus garvieae isolate

Gram-positive cocci recovered from diseased rainbow trout from a farm in England were characterized by different methods, including pulsed field gel electrophoresis, as virulent Lactococcus garvieae serogroup 2 (pulsotype A1). Groups of rainbow trout were kept at a range of temperatures and injected intraperitoneally (i.p.) with one of the UK isolates, L. garvieae 00021. The 18 °C and 16 °C groups showed 67% and 28% mortality, respectively, by day 27 post-injection. Fish kept at 14 °C or lower were less susceptible (≤3% mortality). Raising the temperature of all groups to 18 °C at day 27 post-injection did not result in recurrence of the disease, even though viable bacteria were recovered from all groups 42 days later. Grayling were highly susceptible, with 65% mortalities when challenged with 200 colony forming unit fish⁻¹ by i.p. injection and 37% mortalities when exposed to effluent water from tanks containing affected rainbow trout. Other fish species tested, Atlantic salmon, brown trout and seven cyprinid species, were less susceptible. Viable L. garvieae was isolated from the internal organs of all species tested at the end of the trials, suggesting that they may pose a threat as possible carriers to susceptible farmed and wild fish.     

Thermal tolerance of a thermally selected strain of rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> and the pedigrees of its F1 and F2 generations indicated by their critical thermal maxima

A thermally selected strain of rainbow trout has been established by selective breeding since 1966 in Miyazaki, Japan. In the present study, we compared the critical thermal maxima (CTMs), the temperatures at which organisms reach a predefined sublethal endpoint and lose their equilibrium, between a thermally selected and two normal (Donaldson) strains of rainbow trout. The CTM of one normal strain from Nikko (Nikko strain) acclimated to 20 °C (29.7 °C) was significantly lower than those of the thermally selected strain (30.0 °C) and the other Donaldson strain from Aomori (29.9 °C) (P?<?0.05). The F1 generations, F1T and F1N, were produced by crossing thermally selected strain females with Nikko strain males and Nikko strain females with thermally selected strain males, respectively. No significant difference was observed in the CTM between F1T [30.1?±?0.15 °C (n?=?30)] and F1N [30.1?±?0.16 °C (n?=?30)] (P?>?0.05) for fish acclimated to 20 °C, suggesting that the F1 offspring inherited the thermal tolerance trait from one thermally selected strain parent irrespective of whether it was the male or female. F2 offspring of F1T or F1N also showed the thermal tolerance trait. The coefficients of variation for CTM were also compared among all the datasets obtained in the present study and their values for F1 hybrids were lower than those of the parental generation of the Nikko strain (P?<?0.05). In contrast, the coefficients of variation of F2s were the same as those of their parental generation. Furthermore, the thermally selected strain and Nikko strain as a reference family provide a F2 generation for segregating phenotypes, which is required for in-depth genetic analysis of the thermally selected rainbow trout strain.     

Effects of temperature, irradiance, salinity and inorganic nitrogen concentration on coral zooxanthellae in culture

SUMMARY: Effects of temperature, irradiation, salinity and inorganic nitrogen concentration on two cultured strains of zooxanthellae isolated from the corals Pocillopora damicornis (strain P) and Montipora verrucosa (strain M) were studied. Each strain showed different growth responses in terms of temperature and light intensity. A maximum growth rate of strain P, 1.2 per day, was observed at 32°C under all light intensities examined (5–40 μEm⁻²s⁻¹). However, its photosystem 2 activity (FRI) was higher at 28°C than at 32°C under most light intensities. In contrast, the growth rate of strain M was affected more by light intensity and was almost invariably affected at all temperatures examined (24–36°C). Both algal strains had a comparable growth rate and FRI at salinities from 20 to 35 PSU under moderate temperature and irradiant conditions. High temperature and low irradiation reduced the algal tolerance against low salinity. The gross photosynthesis per cell was not affected by the ammonium enrichment more than 5 μM per day although the cellular chlorophyll a content and cell density increased in proportion with the ammonium enrichment up to 20 μM per day. A potential response of zooxanthellae to the multiple environmental stresses was shown from these results.     

Temperature-dependent index of mitotic interval τ0 in greenling Hexagrammos otakii

ABSTRACT:   The aim of this study was to establish effective procedures for chromosome manipulation in greenling Hexagrammos otakii Jordan et Starks, which has enormous aquacultural potential. To accomplish this, temperature-dependent measurements of the mitotic intervals (τ₀) were carried out. The τ₀ in this fish was determined by averaging the duration of the first and third embryonic divisions at temperatures ranging 5–25°C. At higher temperatures, eggs developed faster and underwent more identical development. For greenling, τ₀ were 341.1 ± 3.60 min at 5°C, 275.5 ± 4.53 min at 10°C, 189.7 ± 6.93 min at 15°C, 99.2 ± 8.27 min at 20°C and 34.2 ± 8.74 min at 25°C. There were strong, negative correlations between the τ₀ and water temperatures at all temperatures studied ( Y  = −79.3 X  + 425.3, R ² = 0.9968, where Y is the mitotic interval and X is the temperature).     

Effects of water temperature on early development of Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

ABSTRACT:   To determine an optimal temperature range for efficient production of healthy eel larvae Anguilla japonica , the effect of water temperature on hatching, survival, and deformity rates was examined. The early ontogeny of morphological features in this species by incubating eggs at five different temperatures (19, 22, 25, 28 and 31°C) was examined. Hatching occurred at 24, 28, 46 and 58 h after fertilization when incubated at 28, 25, 22 and 19°C, respectively; no eggs hatched at 31°C. The growth rate of prefeeding larvae increased as water temperature was elevated and acquisition of feeding ability was also accelerated at higher temperature. Significantly high hatching rates (76–86%, P  < 0.05) and survival rates (61–86%, P  < 0.05) were observed at relatively high temperatures (22–28°C). The deformities were 'open lower jaw', 'pericardial edema', and 'notochordal bending', in which the mouth was deformed with a downward projecting lower jaw, the pericardial cavity was swollen, and the notochord was bent or twisted to various degrees, respectively. Open lower jaw and pericardial edema were especially predominant at 19°C, with rates of 68 and 92%, respectively, compared with 31 and 10% at 25 and 28°C, respectively. The occurrence of notochordal bending was not affected by temperature. The optimal temperature for incubation and rearing A. japonica eggs and prefeeding larvae is approximately 25–28°C.     

Growth and Survival of Intrastrain and Interstrain Rainbow Trout (Oncorhynchus mykiss) Triploids1

Triploid rainbow trout exhibit improved survival and extended growth during sexual maturation, compared to their diploid counterparts. However, there have been few benefits demonstrated prior to sexual maturation. This study was undertaken to investigate the possibility of improving growth and survival parameters in triploids through interstrain crosses. Triploids were induced by heat shocking fertilized eggs from intra- and interstrain crosses of two rainbow trout strains. The four triploid groups and their diploid counterpart groups were reared to 233 days post-hatching and analyzed for growth and survival characteristics. Compared with diploids, triploids had significantly (P < 0.05) higher mortalities during the first 100 days post-fertilization, primarily just prior to and after hatching. However, during the remainder of the study triploids exhibited significantly (P < 0.05) lower mortalities than diploids. During the first 50 days of rearing all four triploid groups were significantly shorter and lighter than their diploid counterparts. The growth of the triploid groups later in the study varied considerably. At the conclusion of the rearing phase, one interstrain triploid group was significantly (P < 0.05) longer than its diploid counterpart, although not significantly heavier. The other triploid groups were either significantly smaller than, or equal to the diploids. Analysis of variance indicated that the growth of triploid rainbow trout was significantly affected by maternal strain effects. These results suggest that the use of specific strains and crosses may improve the growth of triploid rainbow trout.     

Effect of different permeable and non-permeable cryoprotectants on the hatching rate of rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) embryos

The purpose of the present study was to investigate the toxicity of cryoprotectants on the hatching rate of rainbow trout (Oncorhynchus mykiss) embryos. Epiboly and first eye pigmentation stage embryos were immersed in six permeable cryoprotectants, dimethyl sulfoxide (DMSO), glycerol (Gly), methanol (MeOH), propylene glycol (PG), ethylene glycol (EG), and acetamide (Ac), in concentrations of 1–5 M for 5 or 10 min and two non-permeable cryoprotectants, sucrose (Suc) (10%, 15%, 20%) and polyvinyl pyrrolidone (PVP) (5%, 10%, 15%) for 5 min. The embryos were then washed and incubated until hatching occurred. The toxicity of the cryoprotectant was assessed by the hatching rate. The results illustrated that permeable cryoprotectant toxicity for rainbow trout embryos increased in the order of PG < DMSO < MeOH < Gly < EG < Ac. The hatching rate of the embryos treated with permeable cryoprotectants decreased (P < 0.05) with increased concentration and duration of exposure. There were no significant decreases in hatching rate of embryos treated with sucrose and PVP with the increase of concentration; sucrose had higher hatching rates than PVP. Rainbow trout embryos at first eye pigmentation stage exhibited greater tolerance to cryoprotectants than embryos at epiboly stage.     

Egtved virus: temperature-dependent immune response of trout to infection with low-virulence virus

Abstract. The present report deals with experiments concerning the theoretical basis for development of a live, attenuated vaccine against viral haemorrhagic septicaemia (VHS) of rainbow trout, Salmo gairdneri Richardson, on the basis of a virus strain derived from the Egtved virus reference strain F, the pathogenicity of which has been strongly reduced by in vitro passage. It is shown that the low pathogenicity of the virus (designated the Reva strain) is a genetically stable feature and that the protection against VHS induced by infection with the virus is due to an immune response. Following immunological priming at 10°C partial protection developed at 5° 10° and 15° but not at 20°C, whereas neutralizing antibody was produced at all four temperatures by some of the fish. The duration of virus persistence in trout was found to be inversely proportional to the water temperature.     

Effects of temperature changes on the endogenous rhythm of oxygen consumption in the Japanese flounder <Emphasis Type="Italic">paralichthys olivaceus</Emphasis>

ABSTRACT:   The effects of temperature changes on the endogenous rhythm of oxygen consumption in the Japanese flounder Paralichthys olivaceus which were acclimated to 3, 14 or 20°C in advance were investigated. The oxygen consumption rate (OCR) of individuals (total length >28 cm) that were acclimated to 3°C, maintained a relatively low value of 6.9 ± 2.3 mL (mean ± SD) O₂/kg ww (wet weight)/h till 5.8°C, which is considered a 'minimum' rate. Thereafter, the OCR gradually increased up to 18.7 mL O₂/kg ww/h at the temperature of 10.6°C. The OCR of individuals acclimated to 14°C stabilized with 124.2 ± 24.1 mL O₂/kg ww/h, when tested at the relatively constant temperature of 14.4 ± 0.17°C. Maximum entropy spectral analysis revealed that the peak OCR occurred at 23.9 h intervals, which could be a circadian rhythm. The mean OCR showed a peak at 06.00–08.00 hours. For the temperature above 25°C in which the fish were acclimated to 20°C, the rhythmic patterns of OCR were observed until the experimental temperature reached 26.4°C. For the temperature above 28.4°C, the metabolic activity rhythm of the OCR was dampened and then the OCR abruptly rose again. It is assumed that the oscillator of the endogenous rhythm lost the governing of normal metabolic activity in Japanese flounders above 28.4°C. These results suggest that the temperatures of around 5.8, 26.4 and 28.4°C are the 'lower limit temperature', 'upper incipient lethal temperature', and the 'critical thermal maximum', respectively, for the endogenous rhythm of oxygen consumption in the Japanese flounder.     

Diurnal variation in thermal environment experienced by salmonids in the North Pacific as indicated by data storage tags

Eight temperature-recording data storage tags were recovered from three salmonids in Alaska (pink and coho salmon and steelhead trout) and five chum salmon in Japan after 21–117 days, containing the first long-term records of ambient temperature from Pacific salmonids migrating at sea. Temperature data imply diel patterns of descents to deeper, cooler water and ascents to the surface. Fish were found at higher average temperatures at night, with narrower temperature ranges and fewer descents than during the day. Fish tagged in the Gulf of Alaska were at higher temperatures on average (10–12°C) than chum salmon tagged in the Bering Sea (8–10°C). Chum salmon were also found at a wider range of temperatures (−1–22°C vs 5–15°C). This is probably related both to the different oceanographic regions through which the fish migrated, as well as species differences in thermal range and vertical movements. Proportions of time that individual fish spent at different temperatures seemed to vary among oceanographic regions. Steelhead trout may descend to moderate depths (50 m) and not be limited to the top few metres, as had been believed. Japanese chum salmon may seek deep, cold waters as they encounter warm surface temperatures on their homeward migrations. Temperature data from all fish showed an initial period (4–21 days) of day and night temperatures near those of sea surface temperatures, suggesting a period of recuperation from tagging trauma. A period of tagging recuperation suggests that vertical movement data from short-term ultrasonic telemetry studies may not represent normal behaviour of fish. The considerable diurnal and shorter-term variation in ambient temperatures suggests that offshore ocean distribution may be linked more to prey distribution and foraging than to sea surface temperatures.     

Effects of water discharge and temperature on the seaward migration of anadromous browntrout, Salmo trutta, smolts

Abstract – The smolt run of anadromous brown trout ( Sulmo trutta ) in a Norwegian river was studied for three consecutive years. The main run occurred in a period of 7–10 days in the middle or second half of May. Support was found for the hypothesis that high water discharge and temperature triggered the run. Few smolts descended when the discharge was low (<50 m³· s⁻¹) and the water temperature was below 4°C. The maximum number were caught when the discharge was moderate (70–150 m³· s⁻¹) and the water temperature high (68°C). The relative importance of these factors varied from year to year. The first-year discharge explained 38% of the variation in the number of smolts migrating. Discharge and temperature together explained 61% in 1992 and 28% in the second and third year, respectively.