Storage and preservation of temperate mushroom cultures, agaricus bisporus and pleurotus Florida

Two temperate mushroom cultures namely Agaricus bisporus (U-3) and Pleurotus florida (PAU-5) were evaluated for their physiological (linear growth and biomass production), biochemical (β-1,4 endoglucanase production) and fruiting behaviour after preservation in 10% (v/v) glycerol and storage at room temperature (25–35°C), −20°C and −196°C for 6 months with the objective of establishing the recovery/changes in these fungi after storage. Studies indicated that the viability and recovery of A. bisporus and P. florida is affected by the storage conditions. Both the fungi could be best stored in liquid nitrogen for longer durations but for regular use, conventional sub-culturing was appropriate.     

The effect of temperature on growth,physiology, and gametogenesis in the Manila clam Tapes philippinarum (Adams & Reeve, 1850)

Populations of the Manila clam, Tapes philippinarum (Adams & Reeve, 1850), were maintained at temperatures of 12, 15, 18, and 21°C for a period of 19 weeks. Regular determinations were made of ammonia excretion rate following which animals were sacrificed for estimation of dry meat weight, dry shell weight, biochemical composition, and gonadal development. T. philippinarum increased from an initial dry meat weight of 291.5 mg to final values of 957.9, 733.0, 735.0, and 586.0 mg at 12, 15, 18, and 21 °C, respectively.An initial increase in percentage carbohydrate content was evident at all temperatures. This was followed by carbohydrate depletion associated with gametogenesis, the transition from accumulation to depletion occurring earlier with increasing temperature. Ripe gonadal material was evident at all temperatures, but spawning was only evident at 15, 18, and 21 °C. Ammonia excretion at 12, 15, and 18°C showed an allometric relationship to meat weight; this was not so at 21 °C.     

Biochemical composition of temperate and Arctic populations of Saccharina latissima after exposure to increased pCO2 and temperature reveals ecotypic variation

Previous research suggested that the polar and temperate populations of the kelp Saccharina latissima represent different ecotypes. The ecotypic differentiation might also be reflected in their biochemical composition (BC) under changing temperatures and pCO₂. Accordingly, it was tested if the BC of Arctic (Spitsbergen) and temperate S. latissima (Helgoland) is different and if they are differently affected by changes in temperature and pCO₂. Thalli from Helgoland grown at 17 °C and 10 °C and from Spitsbergen at 10 °C and 4 °C were all tested at either 380, 800, or 1,500 µatm pCO₂, and total C-, total N-, protein, soluble carbohydrate, and lipid content, as well as C/N-ratio were measured. At 10 °C, the Arctic population had a higher content of total C, soluble carbohydrates, and lipids, whereas the N- and protein content was lower. At the lower tested temperature, the Arctic ecotype had particularly higher contents of lipids, while content of soluble carbohydrates increased in the Helgoland population only. In Helgoland-thalli, elevated pCO₂ caused a higher content of soluble carbohydrates at 17 °C but lowered the content of N and lipids and increased the C/N-ratio at 10 °C. Elevated pCO₂ alone did not affect the BC of the Spitsbergen population. Conclusively, the Arctic ecotype was more resilient to increased pCO₂ than the temperate one, and both ecotypes differed in their response pattern to temperature. This differential pattern is discussed in the context of the adaptation of the Arctic ecotype to low temperature and the polar night.     

Effects of ambient temperature and of temperature acclimation on nitrogen excretion and differential catabolism of protein and nonprotein resources in the intertidal snails,Littorina saxatilis (Olivi) and L. obtusata (L.)

Nitrogenous excretion in two snails, Littorina saxatilis (high intertidal) and L. obtusata (low intertidal) was studied in relation to temperature acclimation (at 4° and 21°C), including total N excretion rates, the fraction of urea in N excretion, corresponding O:N ratios and the partitioning of deaminated protein between catabolic and anabolic processes at 4°, 11° and 21°C. Aggregate N excretion rates in both species showed no significant compensatory adjustments following acclimation. Total weight specific N excretion rates at 21°C were higher in standard 3 mg L. saxatilis (739 ng N mg⁻¹ h⁻¹) than standard 5 mg L. obtusata (257 ng N mg⁻¹ h⁻¹) for snails acclimated to 21°C. Comparisons of Q₁₀ values of total weight specific N excretion to Q₁₀ values for weight specific oxygen consumption ({xxV}O₂) between 4° to 11 °C and 11° to 21°C indicated that, while total rates of catabolic metabolism ({xxV}O₂) and protein deamination in L. obtusata were essentially parallel, the relationship between N excretion and {xxV}O₂ in L. saxatilis revealed the partitioning of a larger share of deaminated protein carbon into anabolism at 4° and 21°C than at 11°C. Urea N accounted for a larger share of aggregate N excreted in L. saxatilis than in L. obtusata, but in both species urea N is a greater proportion of total N excreted when acclimated at 4°C (urea N: ammonia N ratio range: 1 to 2.15) than in snails acclimated to 21°C (urea N: ammonia N ratio range: 0.46 to 1.39). Molar O:N ratios indicate that the proportion of metabolism supported by protein catabolism is greater in L. saxatilis (O:N range: 2.5–8.4) than in L. obtusata (O:N range: 7.3–13.0). In both species, regardless of acclimation temperature, the O:N ratios are generally lowest (high protein catabolism) at 4°C and highest at 21°C.     

The combined effect of temperature and salinity changes on osmoregulation and haemocyanin concentration in Saduria entomon (Linnaeus, 1758)

Global warming is having an impact on the temperature and salinity of Baltic Sea waters. Therefore, it is important to determine the conditions in which animals can exist and how these changes may influence their functioning. Hence, the purpose of this research was to determine the broad tolerance limits of temperature and salinity of the glacial relict Saduria entomon by studying its behaviour, osmoregulatory ability and haemocyanin concentration. This effect of temperature was confirmed in the laboratory for individuals acclimated to different salinity and temperature regimes. Changes in the physiological parameters of S. entomon at various temperatures (5.5–21.5°C) and salinity levels (1–15 PSU) were recorded. There were statistically significant differences in haemolymph osmotic pressure under the influence of salinity and temperature. The mean haemolymph osmotic pressures were the lowest at 1 PSU at all the temperatures examined and the highest at 15 PSU and high temperatures 16.5 and 21.5°C. The haemocyanin concentration decreased significantly with increasing temperature at 1 PSU. There was a significant difference in haemocyanin concentration due to salinity at temperatures of 5.5 and 10.0°C (the haemocyanin concentration decreased with increasing salinity). The results showed that, although S. entomon is classified as a cold-water animal, it can survive at high temperatures above 16.5°C at least for a short time, as it is capable of osmoregulation. The tolerance to temperature changes was better than expected.     

Effect of Temperature and Salinity on the Biochemical Composition of Artemia franciscana KELLOGG 1906, from Araya (Venezuela) and San José (México)

The effect of different temperature and salinity combinations on the biochemical composition of Artemia franciscana from Venezuela and Mexico, is analyzed. Temperatures were 22 ± 0.5 °C, 26 ± 0.5 °C and 30 ± 0.5°C; salinities were 30‰, 60‰, and 120‰. Chaetoceros sp. was used as food. According to Tukey's Multiple Range Analysis for the A. franciscana population from Araya and San José, there were differences in the biochemical parameters and survival percentages among treatments and between populations. A positive correlation is observed among proximate composition values and survival, total length and growth rates. The observed variations reflect a genetic component resulting from the life history of the populations, and a non-genetic component produced by the experimental conditions.     

An X-ray diffraction study on phase transition temperatures of various membranes isolated from Tetrahymena pyriformis cells grown at different temperatures

Mitochondrial, microsomal and pellicular membranes were isolated from Tetrahymena cells grown at 39°C or 15°C, and phospholipids, in turn, were separated from total lipids extracted from these membranes. The effect of growth temperature on their solid-to-fluid phase transition temperature was examined by wide-angle X-ray diffraction. The transition temperatures of phospholipids from mitochondria, microsomes and pellicles were 21, 19 and 26°C for cells grown at 39°C and ?8, ?3 and 6°C for cells grown at 15°C, respectively. All phospholipids were found in a completely fluid state at these growth temperatures. From a comparison between the phospholipids and total lipids from pellicles of cells grown at 39°C, a triterpenoid alcohol, tetrahymanol, caused the transition temperature to increase. The alignment of tetrahymanol in membranes was examined with pellicle's total lipid oriented in a sample holder.     

THERMAL EVOLUTION OF EGG SIZE IN DROSOPHILA MELANOGASTER

We measured the size of eggs produced by populations of Drosophila melanogaster that had been collected along latitudinal gradients in different continents or that had undergone several years of culture at different temperatures in the laboratory. Australian and South American populations from higher latitudes produced larger eggs when all were compared at a standard temperature. Laboratory populations that had been evolving at 16.5°C produced larger eggs than populations that had evolved at 25°C or 29°C, suggesting that temperature may be an important selective agent in producing the latitudinal clines. Flies from laboratory populations produced larger eggs at an experimental temperature of 16.5°C than at 25°C, and there was no indication of genotype-environment interaction for egg size. Evolution of egg size in response to temperature cannot be accounted for by differences in adult body size between populations. It is not clear which life-history traits are direct targets of thermal selection and which are showing correlated responses, and disentangling these is a task for the future.     

Enhancement of Thermal Stabilization of Formaldehyde Dehydrogenase from Pseudomonas putida by Directed Evolution

We used directed evolution to enhance the thermostability of formaldehyde dehydrogenase from Pseudomonas putida. At 50 °C, the wild-type enzyme was inactivated within 30 min, but the variants obtained retained 80% activity for at least 300 min. At room temperature (30 °C), the variants obtained retained <80% activity for at least 500 h (21 d).     

RESPONSE OF TRACHYDISCUS MINUTUS (XANTHOPHYCEAE) TO TEMPERATURE AND LIGHT1

The effects of different temperatures and light intensities on growth, pigments, sugars, lipids, and proteins, as well as on some antioxidant and proteolytic enzymes of Trachydiscus minutus (Bourr.) H. Ettl, were investigated. The optimum growth temperature and light intensity were 25°C and 2 × 132 μmol photons · m^?2 · s^?1, respectively. Under these conditions, proteins were the main biomass components (33.45% dry weight [dwt]), with high levels of carbohydrates (29% dwt) and lipids (21.77% dwt). T. minutus tolerated temperatures between 20°C and 32°C, with only moderate changes in cell growth and biochemical composition. Extremely low (15°C) and high (40°C) temperatures decreased chl and RUBISCO contents and inhibited cell growth. The biochemical response of the alga to both unfavorable conditions was an increase in lipid content (up to 35.19% dwt) and a decrease in carbohydrates (down to 13.64% dwt) with much less of a change in total protein content (in the range of 30.51%–38.13% dwt). At the same time, the defense system of T. minutus was regulated differently in response to heat or cold treatments. Generally, at 40°C, the activities of superoxide dismutase (SOD), catalase (CAT), and proteases were drastically elevated, and three polypeptides were overexpressed, whereas the glutathione reductase (GR) and peroxidase (POD) activities were reduced. In contrast, at 15°C, all these enzymes except GR were suppressed. The effect of light was to enhance or decrease the temperature stress responses, depending on intensity. Our studies demonstrate the broad temperature adaptability of T. minutus as well as the potential for the production of valuable algal biomass.     

The effect of different preservation methods on the carbon content of megacyclops gigas

The effect of various preservation methods on the carbon content of the copepod Megacyclops gigas was studied using the carbon content of living animals as a control. No differences were found between animals dried at –20, +60, or +105°C, but in all cases the carbon content was slightly lower than that found in living animals. Chemical preservation with formaldehyde and acid Lugol's solution rapidly resulted in about 35% loss of carbon. Freezing at –20°C in water caused less marked but significant loss of carbon, whereas freezing in dilute formaldehyde yielded values agreeing with those of living animals.     

Effect of temperature on antioxidant enzymatic activity in the Pacific white shrimp Litopenaeus vannamei in a BFT (Biofloc technology) system

A 10-day trial was conducted to evaluate the effect of temperature on the antioxidant system of Litopenaeus vannamei in a biofloc technology system. Four treatments in triplicate tanks were assigned to the following temperatures: 15, 21, 27 (control), and 33 °C and the water quality parameters were monitored every day. For all enzyme assays, the hemolymph cells lysate of six shrimp per treatment was collected. Lipid peroxidation (LPO) was assessed by determining the content of thiobarbituric acid-reactive substances (TBARS); the activities of catalase (CAT) and glutathione-S-transferase (GST) were also evaluated. The results of TBARS showed that shrimp reared at 15 and 21 °C presented an increase of 407% (240 h) and 339% (120 h), respectively. L. vannamei exposed to 15 °C augmented in 186% (24 h) the activity of CAT. Moreover, shrimp of 21 °C group also increased CAT activity in 228% (6 h). GST presented the strongest variation reaching 1437% in shrimp of 15 °C for 6 h and 1425% of the 21 °C treatment at the same time. All these results were compared to the initial time (27 °C). Moreover, the antioxidant system was not sufficient to counteract LPO; therefore, the animals reared in 15 and 21 °C presented higher enzyme activities, suggesting that cold water can induce the oxidative stress in L. vannamei.     

Body reserve dynamics and energetics of barn owls during fasting in the cold

The balance of protein versus lipid reserves utilization in fasting animals depends on their initial adiposity, a high prefasting adiposity being associated with an efficient protein sparing during fasting. Yet it remains unclear if the level of energy expenditure influences the efficiency of body protein sparing. We examined the effect of a high energy demand on body reserve mobilization in barn owls (Tyto alba) fasting in the cold (5 °C). Changes in body composition of captive birds were determined during the three characteristic phases of body fuel utilization of a long-term but reversible fast. Although showing a low prefasting adiposity (12%), barn owls spared body protein in phase II as efficiently as the fattest species (contribution to energy expenditure of <9%). This low protein utilization most probably results from an increased lipid mobilization in the cold. This argues for an influence of a high energy demand on the relative efficiency of protein sparing. For lipids, the pattern of mobilization from tissue sources is similar in barn owls to that of species fasting at thermoneutrality. For proteins, in contrast, and despite a low decrease of the total body protein mass (16%), digestive tract and liver were affected most, with respective losses of 43% and 62% at phase III. This could be another consequence of the interaction between high energy demand in the cold and fasting. Indeed, high cold-induced thermoregulatory needs could result in selective preservation of organs involved in the thermoregulatory process (muscles) to the detriment of lesser solicited organs such as those involved in digestion. Accepted: 8 September 1999     

The effect of temperature on the growth,development and survival of Macropetasma africanus (Balss) (Penaeoidea:Penaeidae) larvae reared in the laboratory

Macropetasma africanus (Balss) has been successfully spawned and its larvae reared under controlled laboratory conditions. The relationship between egg number (E) and female total length (L) was E = 18.59 L^2.11. An experiment was designed to test the effect of temperature on larval development, survival and growth. Temperature effected larval development time, from 13–15 days at 25°C, to 25 days at 15°C (nauplius 1 to post-larva). Mortality was low for the naupliar stages at 25, 22 and 18°C, while at 15°C only 52% of the larvae reached nauplius 6. Mortality was highest from nauplius 6 to protozoea 1 (17, 21, and 18% at 25, 22, and 18°C, respectively), but decreased considerably for all temperatures once the mysis stage was reached. Overall survival rates from nauplius 1 to post-larva decreased with decreasing temperature (65, 54, 48, and 39% at 25, 22, 18, and 15°C respectively). Temperature also significantly affected larval growth. At 25°C mean total length was significantly (P < 0.05) larger than at 15°C (protozoea 2 to post-larva), while from protozoea 3 to post-larva total length differences were significantly different (P < 0.05) between 18 and 25°C. M. africanus has a major spawning peak in summer, suggesting that there may be a selective advantage to reproducing during the warmer months.     

Sex- and age-related changes in the biophysical properties of cuticular lipids of the housefly,Musca domestica

We examined the biophysical properties of cuticular lipids isolated from the housefly, Musca domestica. Melting temperatures (T_m) of surface lipids isolated from female houseflies decreased from 39.3 °C to 35.3 °C as the females attained sexual maturity and produced sex pheromone, whereas those prepared from males did not change with age. Lipids melted over a 10–25 °C temperature range, and their physical properties were a complex function of the properties of the component lipids. The T_m of total cuticular lipids was slightly below that of cuticular hydrocarbons (HC), the predominant lipid fraction. Hydrocarbons were further fractionated into saturated, unsaturated, and methyl-branched components. The order of decreasing T_m was total alkanes > total HCs > methyl-branched alkanes > alkenes. For 1-day-old flies, measured T_ms of hydrocarbons were 1.3–5.5 °C lower than T_ms calculated from a weighted average of T_ms for saturated and unsaturated components. For 4-day-old flies, calculated T_ms underestimated T_m by 11–14 °C. © 1995 Wiley-Liss, Inc.     

Changes in selected aspects of immune function in the leopard frog, Rana pipiens, associated with exposure to cold

The effect of exposure to low temperatures (5 °C) on lymphocyte proliferation, leukocyte populations, and serum complement levels was examined in the northern leopard frog, Rana pipiens. Proliferation of T lymphocytes in response to phytohemagglutinin stimulation was significantly decreased in frogs kept for 2, 3, and 5 months at 5 °C compared to that of animals kept at 22 °C. A significant increase in the average percentage of neutrophils and a decrease in the mean percentage of eosinophils was observed in the blood of frogs held for 5 months in the cold compared to animals held at 22 °C for the same length of time. Mean serum complement activity after 1 month at 5 °C was significantly reduced in comparison to animals held at 22 °C and was not detectable after 5 months in the cold. Recovery of complement levels at room temperature (22 °C) was also examined after cold exposure. Complement levels were significantly higher than controls (at 22 °C) in frogs returned to 22 °C for 7 and 14 days after 5 months in the cold. After frogs were held at 5 °C for 1 month, serum complement levels increased significantly within 2 days after returning to 22 °C and continued to rise 5 and 9 days after warming. Injections with Aeromonas hydrophila following a 5-week exposure to 5 °C failed to cause death or observable symptoms of disease in frogs that were returned to 22 °C. Accepted: 20 November 1996     

Preserving young‐of‐the‐year Perca fluviatilis in ethanol,formalin, or in a frozen state and the consequences for measuring morphometrics

As it is often not possible to immediately analyse individuals sampled in the field, captured fish are preserved and stored for later investigation. The objective of this study was to assess the effects of various preservation methods on subsequent changes in morphometric parameters while also providing correction factors to re‐calculate the original body dimensions when sampled fish are measured at a later date. In this study, juvenile perch (Perca fluviatilis, 66.5 ± 10.2 mm total length) were measured directly after capture, then either frozen at ?20°C, preserved in 70% ethanol, or in 4% formalin. They were again measured after 24 h, 3 days, 7 days, and thereafter on a weekly basis for 8 weeks. Ethanol‐preserved perch greatly decreased length and weight; formalin preservation also led to a comparable length reduction, but increased the weight of the perch. In contrast, frozen perch showed less shrinkage and only moderate weight loss. Of the three preservation methods, freezing clearly caused the fewest distortions. Hence, freezing is recommend as the most preferable preservation method, especially in multi‐disciplinary studies on fish ecology; for all other preservation methods the correction factor over time will have to be pre‐determined for each species and size class.     

Growth and maturation of a North American fairy shrimp, Streptocephalus seali (Crustacea; Anostraca): a laboratory study

SUMMARY.

• 1 We evaluated survival, growth and time to maturation of the fairy shrimp, Streptocephalus seali Ryder, in the laboratory at various combinations of temperature and water hardness.
• 2 Both independent factors affected survival and growth of S. seali. Multiple regression analysis and response surface modelling predict that after 4 days, over 80% survival is obtained at temperatures from 14 to 28°C and water hardnesses from 60 to 130 mg CaCO₃ 1-^?1.
• 3 Growth rates of larvae were often maximum at physicochemical conditions other than those which had promoted maximum rates of survival. For example, after 12 days mean total body length was almost 12 mm in larvae which had been maintained at 34°C (80 mg CaCO₃ 1-^?1): the maximum survival rate had been obtained at 19°C. Total length was directly correlated with temperature at the lowest hardness tested, but not at the other two hardnesses (100 and 120 mg CaCO₃ 1-^?1). At the latter water hardnesses, total length was significantly less at 34°C than at 32°C on all three sampling occasions (4, 8 and 12 days post-hatch).
• 4 Similarly, developmental stage of larvae correlated well with temperature but larvae reared at 34°C did not develop more quickly than those reared at 32°C. After 12 days, most larvae at the two highest temperature treatments had developed at least to Stage 14 and many were nearly mature; at 17°C most larvae were still at Stage 10.
• 5 During our study of maturation rate of females we noted that egg production was initiated after completion of fourteen or fifteen moults. Mean time to maturation at 27°C (17.3±2.8 days) exceeded that at 32°C (12.3±2.6days). The minimum time to maturation of a shrimp was 9 days at 32°C.

     

Thermal evolution of rate of larval development in Drosophila melanogaster in laboratory and field populations

The duration of Drosophila melanogaster larval and pupal periods was measured in laboratory thermal lines and in populations collected along a latitudinal transect in eastern Australia. In replicated laboratory lines kept for 9 years at 16.5° C or 25° C the duration of larval development had continued to diverge compared with 4 and 5 years previously, with more rapid larval development, and hence reduced total duration of pre-adult development, in the low temperature lines at both experimental temperatures. After 4 years of separate evolution, lines derived from the 25° C lines and subsequently cultured at 29° C showed no evidence of significant divergence in the duration of any part of the pre-adult period. The geographic populations showed a decrease in the duration of larval development, and hence of the total pre-adult period, with increasing latitude. In both laboratory and field populations, evolution at lower temperature was associated with more rapid larval development to a larger adult body size, the opposite genetic correlation between these traits to that found within a single temperature. The indications are that lower temperatures may be permissive of more efficient growth in D. melanogaster. It will be important to discover if evolution in response to temperature induces similar correlations in other ectotherms.