THE EFFECT OF TEMPERATURE ON EGGS AND IMMATURE STAGES OF CULEX ANNULIROSTRIS SKUSE (DIPTERA: CULICIDAE)

Abstract
No immature stages of Culex annulirostris were found during field sampling in 1979–1980 when the average water temperature was < 17 °C; they reappeared when the average water temperature was 19 °C and reached the peak density (mean 107 immatures/cylinder) at 26.5 °C.
The effect of 6 temperatures (15–40°C) on egg hatching, development and survival of the immature stages of Cx annulirostris in the laboratory showed that at 15 and 40°C, eggs failed to hatch and larvae died in the first instars. The optimum temperatures for egg hatching and the survival of immature stages were 25 and 30°C. At these temperatures, 85 and 82% respectively of egg rafts hatched, the mean number of larvae per raft was 258 ± 9.8 and 260 ± 11.4 with immature survival of 83.5 and 79.0% respectively. Mean time to hatch at 20–35°C ranged from 1.2 d (35°C) to 2.9 d (20 °C). Developmental times from first instar to adult ranged from 7.1 d (35 °C) to 25.2 d (20 °C). The threshold for development of the immatures was 15.6 ± 2.5°C and the thermal constant was 142.9 ± 26.5 day—degrees (incubation temperatures 20–35°C). At less suitable temperatures of 20 and 35 °C, hatching (57.5 and 45%), number larvae per raft (mean 139.8 ± 9.8 and 102.6 ± 14.2) and survival were low.     

Growth rate and physiology of Yersinia enterocolitica; influence of temperature and presence of the virulence plasmid

The effect of temperature on the growth rate, protein pattern and fatty acid composition of Yersinia enterocolitica strain W22703 pYV⁺, its plasmidless isogenic derivative W22703 pYV^- and four recent field isolates was examined.
The growth rate was clearly influenced by presence or absence of the virulence plasmid: pYV^- strains grew consistently faster than pYV⁺ strains. This difference in growth rate was high at 30–35°C, moderate at 1–10°C and 25°C, but hardly significant at 15–20°C.
Increasing the growth temperature above 25°C resulted in the induction of the 220 kDa virulence plasmid-encoded Yop1 protein. In the 1–20°C range no obvious temperature- or plasmid-related differences in protein patterns could be detected.
The fatty acid composition showed a clear temperature-dependent change: with all strains the degree of saturation was low at 1°C and gradually increased with raising temperatures. All strains had similar fatty acid patterns, except one of the field isolates which showed aberrant C16 : 1 and cyclic fatty acid contents in the 5–25°C and 15°C ranges respectively. With strain W22703, the presence or absence of the virulence plasmid did not significantly alter the fatty acid pattern.     

Genetic analysis of adenovirus type 2. I. Isolation and genetic characterization of temperature-sensitive mutants.

Temperature-sensitive mutants which replicate normally at 33 C but poorly at 39 C were isolated from nitrosoguanidine- or nitrous acid-mutagenized adenovirus 2 by (i) testing the cytopathic effect or inclusion body-forming capacity of random plaque isolates, or (ii) reduced plaque enlargement upon shifting from 33 to 39 C. Thirty-six mutants were isolated with 33 C/39 C plaque ratios varying from 20 to 10-5. Some of these mutants could be arranged into 13 groups by the complementation test. By means of recombination analysis a provisional linear genetic map was constructed.     

Mu dI1(Ap lac) mutagenesis of Yersinia pestis plasmid pFra and identification of temperature-regulated loci associated with virulence

The F1 capsule of Yersinia pestis, encoded by the 100 kb plasmid pFra, is often assumed to be essential for full virulence of Y. pestis. However, virulent strains of Y. pestis that are F1- and either pFra+ or pFra- have been reported. To assess the role of pFra-encoded factors in virulence, mutants in pFra with insertions of the defective transposing bacteriophage Mu dl(Ap lac) were obtained, by using the wild type (wt) and the pLcr-cured derivative of strain C092. Mutants that exhibited temperature regulation of lactose fermentation and retarded electrophoretic mobility of pFra were selected. A total of 15 insertion mutants were isolated in the wt strain (12 of which had a single insertion in the genome, in pFra); and 24 mutants in the isogenic pLcr- derivative. Four of the pLcr+ mutants, and none of the pLcr- mutants, were F1-. All F1- mutants were decreased in virulence for mice compared to the wt parent; and five of the F1+ mutants also were significantly attenuated in mice. Fusion end-joints of insert DNA were cloned into Escherichia coli by using pMLB524, a vector for rescuing operon fusions of lacZ. Recombinants were obtained which contained pFra inserts ranging from < 2kb to approximately 36 kb, and the insertions occurred at several sites on pFra. All of the four F1- mutants tested mapped within the F1 capsule operon (caf1). The remaining five attenuated mutants sequenced were F1+ and mapped outside of but near the operon. Sequencing and complete analysis of the pFra insertions mutants could facilitate identification of new potential virulence factors.     

Role and mechanism of phosphatidylinositol-specific phospholipase C in survival and virulence of Cryptococcus neoformans

Phospholipase B1 (Plb1) is secreted after release from its glycosylphosphatidylinositol anchor and is implicated in initiation and dissemination of infection of the pathogenic fungus, Cryptococcus neoformans . To investigate the role of phosphatidylinositol-specific phospholipase C (PI-PLC) in Plb1 secretion, we identified two putative PI-PLC-encoding genes in C. neoformans var. grubii ( PLC1 and PLC2 ), and created Δ plc1 and Δ plc2 deletion mutants. In Δ plc1 , which expressed less PI-PLC activity than wild type (WT), three major cryptococcal virulence traits, Plb1 secretion, melanin production and growth at host temperature (37°C) were abolished and absence of Plb1 secretion coincided with Plb1 accumulation in plasma membranes. In addition, Δ plc1 cell walls were defective, as indicated by cell clumping and irregular morphology, slower growth and an inability to activate mitogen-activated protein kinase (MAPK) in the presence of cell wall-perturbing agents. In contrast to Δ plc2 , which was as virulent as WT, Δ plc1 was avirulent in mice and exhibited attenuated killing of Caenorhabditis elegans at 25°C, demonstrating that mechanism(s) independent of the 37°C growth defect contribute to the virulence composite. We conclude that Plc1 is a central regulator of cryptococcal virulence, acting through the protein kinase C/MAPK pathway, that it regulates release of Plb1 from the plasma membrane and is a candidate antifungal drug target.     

Further studies of 31 temperature-sensitive mutants of mouse cytomegalovirus: Thermal stability, replication and analysis of temperature-sensitive functions by temperature shift

Abstract A study of 31 temperature-sensitive mutants of mouse cytomegalovirus has indicated that two mutants ( tsm 1, tsm 31) may be defective in immediate-early/early functions, two ( tsm 2, tsm 3) may be defective in early functions and six ( tsm 9, tsm 18, tsm 22, tsm 23, tsm 28, tsm 30) may be defective in early/late functions while the remainder are late function-defective mutants as determined by temperature-shift experimemts. Three mutants ( tsm 1, tsm 2, tsm 3) were more thermostable than wild-type virus while three ( tsm 16, tsm 26, tsm 28) were more thermolabile; the remainder were similar in their thermostability to wild-type virus.     

Influence of growth temperature on the production of extracellular virulence factors and pathogenicity of environmental and human strains of Aeromonas hydrophila

The biochemical properties, virulence for mice and trout, and the extracellular virulence factors at 28° and 37°C of 11 environmental and nine human strains of Aeromonas hydrophila were compared. All the environmental isolates and four of the human group were virulent for trout at 3 x 10⁷ cfu, but only human strains were able to cause death or lesions in mice by the intramuscular route. Extracellular virulence factors such as haemolysins, cytotoxins and proteases were also investigated in supernatant fluids of cultures grown at 28°C and 37°C. The production of haemolysins, caseinases, elastases and growth yields of environmental strains decreased sharply during cultivation at 37°C but cytotoxins were produced to the same extent, or slightly less, than at 28°C. The human strains differed from the environmental strains in response to growth temperatures: protease activity decreased at 37°C, although growth yield was not affected, but more haemolysins and cytotoxins were produced by the virulent strains at this temperature than at 28°C. Sodium caseinate SDS-PAGE of culture supernatant fluids of selected human strains revealed that temperature selectively inhibited the production of certain proteases.     

Effects of Temperature on Germination of Peronospora parasitica Conidia and Infection of Brassica oleracea

The effect of temperature on germination of a South African isolate of Peronospora parasitica , and infection of Brassica oleracea was studied. The optimum condition for germination was 20° C at 100% relative humidity. The percentage germination obtained was 80–98% and 70–80% between 15 and 25° C at 100% relative humidity, after a 12 and 6h incubation period, respectively. Optimum temperature for germ tube growth was also 20° C. The temperature range for maximum infection of seedlings of a highly susceptible cabbage cultivar and subsequent disease development in vitro was 15–25° C and 90–100% infection was achieved after 48 h of incubation. At<15°C and 26–30° C infection percentage was decreased to 40–50% and 35–40%, respectively. No disease incidence was recorded at temperatures above 35° C. A scanning electron microscope study of the infection process showed that penetration of cotyledons by germ tubes was mostly via stomata and occasionally directly through the cuticle. Results are discussed in relation to the need for future studies of P. parasitica in South Africa.     

The control of postharvest decay in table grapes using acetaldehyde vapours

Grapes ( Vitis vinifera L. cv. 'Sultanina') harvested at the end of the 1985–1988 seasons, received postharvest application of acetaldehyde (AA) vapours for 24–40 h. Treatment with AA vapour at 20 °C or 0 °C reduced significantly the decay caused by several fungi: Botrytis cinerea, Rhizopus stolonifer, Aspergillus niger and Alternaria alternata . In grapes treated with 0.5% AA for 24 h, no R. stolonifer was found after 8 days of storage at 20 °C. Treatment with 0.25% AA vapour for 40 h of grapes cv. 'Perlette' inoculated with R. stolonifer reduced the decay by 89%.     

Effects of different incubation temperatures on the yolk‐feeding stage of Eupallasella percnurus(Pallas)

Embryos and yolk‐feeding larvae of lake minnow Eupallasella percnurus were reared at 13, 16, 19, 22 and 25° C with no access to external food. Time from egg activation to first embryonic movements, hatching, filling of swimbladder and final yolk resorption increased with decreasing temperature. At 13° C, c . 40% of larvae were unable to fill their swimbladder. The predicted lower temperature at which development and growth ceased (biological zero, t ₀) was the same for both processes, c . 7·5–10·5° C. There was no ontogenetic shift in the t ₀ value. Temperature coefficients for development ( Q _10dev.) ranged from 2 to 3 at 19–25° C, but were higher in hatched larvae at lower temperatures. Eggs of E. percnurus had a combination of small size, high hydration and low caloric value of fresh matter. Dry mass of larval tissue on yolk, percentage of dry matter in wet matter, and specific growth rate were maximized at 22 and 25° C. At 19–25° C, energy and matter contained in the initial eggs were converted to body tissue most efficiently. Temperatures from 22 to 25° C are considered optimal for E. percnurus embryos and yolk‐feeding larvae and are recommended for their indoor rearing.     

Effect of Iysozyme concentration, heating at 90°C, and then incubation at chilled temperatures on growth from spores of non-proteolytic Clostridium botulinum

The heat treatment necessary to inactivate spores of non-proteolytic Clostridium botulinum in refrigerated, processed foods may be influenced by the occurrence of lysozyme in these foods. Spores of six strains of non-proteolytic Cl. botulinum were inoculated into tubes of an anaerobic meat medium, to give 10⁶ spores per tube. Hen egg white lysozyme (0–50 μg ml^-1) was added, and the tubes were given a heat treatment equivalent to 19·8 min at 90°C, cooled, and incubated at 8°, 12°, 16° and 25°C for up to 93 d. In the absence of added lysozyme, neither growth nor toxin formation were observed. A 6–D inactivation was therefore achieved. In tubes to which lysozyme (5–50 μg ml^-1) had been added prior to heating, growth and toxin formation were observed. With lysozyme added at 50 μg ml^-1, growth was first observed after 68 d at 8°C, 31 d at 12°C, 24 d at 16°C, and 9 d at 25°C. Thus, in these circumstances, a heat treatment equivalent to 19·8 min at 90°C was not sufficient, on its own, to give a 6–D inactivation. A combination of the heat treatment, maintenance at less than 12°C, and a shelf-life not more than 4 weeks reduced the risk of growth of non-proteolytic Cl. botulinum by a factor of 10⁶.     

Influence of water temperature and feeding regime on otolith growth in Anguilla japonica glass eels and elvers: does otolith growth cease at low temperatures?

The influences of water temperature and feeding regime on otolith growth in Anguilla japonica glass eels and elvers were investigated using individuals reared at 5, 10, 15, 20, 25 and 30° C and in fed or unfed conditions at salinity 32 after their otoliths were marked with alizarin complexone (ALC). To eliminate the difficulty of observing the edges of otoliths with optical (OM) or scanning electron (SEM) microscopes, three to 10 individuals were sampled from each tank at 10, 20 and 30 days during the experiment and reared for an additional 10 days at 25° C after their otoliths were marked a second time. Otolith growth and the number of increments were measured using both OM and SEM. Most A. japonica commenced feeding after 10 days at 20–30° C or after 20 days at 15° C, but no feeding occurred at 5 and 10° C. No otolith growth occurred at 5 and 10° C except in two individuals with minimal increment deposition at 10° C. Otolith growth was proportional to water temperature within 15–25° C and not different between 25 and 30° C. At 15, 25 and 30° C, the mean otolith growth rate in fed conditions was higher than in unfed conditions. The number of increments per day was significantly different among water temperatures (0·00–0·01 day⁻¹ at 5 and 10° C, 0·43–0·48 day⁻¹ at 15° C and 0·94–1·07 day⁻¹ at 20–30° C). These results indicated that otolith growth in A. japonica glass eels and elvers was affected by temperature and ceased at ≤10° C under experimental conditions. Hence, future studies analysing the otoliths of wild-caught A. japonica glass eels and elvers need to carefully consider the water temperatures potentially experienced by the juveniles in the wild.     

Heat shock proteins in willow (Salix viminalis)

The heat shock response in three vegetatively propagated clones of Salix viminalis L. was studied. In the clone 78198, synthesis of a total of 58 proteins was induced or increased by heat shock. Of these proteins, 39 were found in both leaves and callus, 8 only in leaves, and 11 only in callus. The number of heat shock proteins differed between the three clones studied. The molecular weights of the heat shock proteins ranged from 18000 to over 94000. The optimal synthesis of heat shock proteins took place at 37–40°C, but several proteins could be induced at 25–30°C. The synthesis of the majority of the proteins present at a normal growth temperature (20°C) was not completely blocked by the heat shock. More than 12 h was needed for the reappearance of the normal protein synthesis pattern after heat shock.     

Oxygen consumption, ammonia excretion and protein use in response to thermal changes in juvenile Atlantic salmon Salmo salar

Experiments were designed to examine the effects of various temperature challenges on oxygen consumption and ammonia excretion rates and protein utilization in juvenile Atlantic salmon Salmo salar . Fish acclimated to 15° C were acutely and abruptly exposed to either 20 or 25° C for a period of 3 h. To simulate a more environmentally relevant temperature challenge, a third group of fish was exposed to a gradual increase in temperature from 15 to 20° C over a period of 3 h ( c. 1·7° C h⁻¹). Oxygen consumption and ammonia excretion rates were monitored before, during and after the temperature shift. From the ammonia excretion and oxygen consumption rates, protein utilization rates were calculated. Acute temperature changes (15–20° C or 15–25° C) caused large and immediate increases in the oxygen consumption rates. When the temperature was gradually changed ( i.e. 1·7° C h⁻¹), however, the rates of oxygen consumption and ammonia excretion were only marginally altered. When fish were exposed to warmer temperatures ( i.e. 15–20° C or 15–25° C) protein use generally remained at pre-exposure (15° C) levels. A rapid transfer back to 15° C (20–15° C or 25–15° C) generally increased protein use in S. salar . These results indicate that both the magnitude and the rate of temperature change are important in describing the physiological response in juvenile salmonids.     

Acclimation is beneficial at extreme test temperatures in the Danube crested newt, Triturus dobrogicus (Caudata, Salamandridae)

Despite many studies demonstrating the effect of acclimation on behavioural or physiological traits, considerable debate still exists about the evolutionary significance of this phenomenon. One of the unresolved issues is whether acclimation to warmer temperature is beneficial at treatment or at more extreme test temperatures. To answer this question, we assessed the effect of thermal acclimation on preferred body temperatures ( T _ps), maximum swimming and running speed, and critical thermal maximum ( CT _max) in the Danube crested newt ( Triturus dobrogicus ). Adult newts were kept at 15 °C (control) and 25 °C (treatment) for 8 weeks prior to measurements. We measured T _ps in an aquatic thermal gradient over 24 h, maximum speeds in a linear racetrack at six temperatures (5–33 °C), and CT _max in a continuously heated water bath. T _ps were higher in newts kept at 15 °C than in those kept at 25 °C. The maximum swimming speed did not acclimate. The maximum running speed at 30–33 °C was substantially higher in newts kept at 25 °C than in those kept at 15 °C. CT _max increased with the treatment temperature. Hence, we conclude that the acclimation response to warm temperature is beneficial not at treatment but at more extreme temperatures in newts.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 90 , 627–636.     

Observations on the thermoregulatory behaviour of juvenile spotted grunter, Pomadasys commersonnii (Haemulidae: Pisces)

The thermal preference of juvenile spotted grunter, Pomadasys commersonnii (Haemulidae), was determined in a horizontal thermal gradient. Temperature preference was tested over a range of 17–31°C at three salinities. The modal temperature preferenda at 5, 12 and 35 ppt were 25°C, 24.5°C and 25°C, respectively. They were not significantly different. The thermal preferendum of this species was found to be between 24°C and 25°C. Supplementary experiments were conducted to test the stability of the final thermal preferendum. Initial observation of the behaviour of juvenile spotted grunter indicated the presence of a strong circadian rhythm. However, diel variation in activity did not influence temperature preference. These data are important for the development of a culture protocol for this euryhaline species.     

Effect of temperature on the biology of Creontiades dilutus (Stål) (Heteroptera: Miridae)

The egg and nymphal development, fecundity and survival of the green mirid, Creontiades dilutus were examined at a range of temperatures and a modified day-degree model fitted to the data. Day degree (DD) requirements for egg and nymphal development, and threshold temperatures were calculated from the fitted lines. Female fecundity and longevity, egg and nymphal development, and survival of C. dilutus were significantly influenced by temperature. Eggs and nymphs failed to complete development at temperatures below 17 and at 38°C. Females also failed to produce any eggs at 11 and 38°C. The optimum temperature range for female fecundity was found to be 26–32°C. The optimum temperature for the development of eggs was calculated from the model as 30.5°C and for nymphs as 31.5°C. The threshold temperature for development was 15.8°C for egg and 15.1°C for nymph; 69.4 and 156.7 DD were required for completing the egg and the nymphal development, respectively. At the optimum temperature, it was estimated that development from egg to adult took 15 days. Survival was highest at 26°C for eggs and at 30–32°C for nymphs.     

Ultra-structure and localisation of formazan formed by human neutrophils and amoebae phagocytosing virulent and avirulent Legionella pneumophila

Abstract Legionella pneumophila (LP) strains of differing virulence were incubated with a solution of nitroblue-tetrazolium (NBT) at a concentration of 1 mg · ml⁻ in the presence of Acanthamoeba polyphaga or human polymorphonuclear neutrophils (PMN). Reduction of NBT to formazan occurred at a faster rate in the presence of virulent strains. Reduction appeared to be temperature dependent; at 37°C the reaction rate was higher than at 20°C. On microscopic examination, deposits of formazan around Legionella cells were observed inside amoebae similar to those deposited in human neutrophils. Electron microscopy revealed electron-dense particles surrounding virulent legionellae, which appeared to be associated with formazan foundation. Formazan formation inside amoebae may suggest the presence of a respiratory burst against LP, which is more intense with virulent strains.