The effect of cold acclimation and deacclimation on cold tolerance,trehalose and free amino acid levels in Sitophilus granarius and Cryptolestes ferrugineus (Coleoptera)

Canadian and French laboratory strains of Sitophilus granarius (L.) and Cryptolestes ferrugineus (Stephens) were cold acclimated by placing adults at 15, 10 and 5 degrees C successively for 2wk at each temperature before deacclimating them for 1wk at 30 degrees C. Unacclimated S. granarius had an LT(50) (lethal time for 50% of the population) of 12days at 0 degrees C compared with 40days after the full cold acclimation. At -10 degrees C, unacclimated C. ferrugineus had an LT(50) of 1.4days compared with 24days after the full acclimation. Cold acclimation was lost within a week after returning insects to 30 degrees C. Trehalose, as well as the amino acids proline, asparagine, glutamic acid and lysine were higher in cold acclimated insects for both species. For S. granarius, glutamine was higher in cold acclimated insects and isoleucine, ethanolamine and phosphoethanolamine, a precursor of phospholipids, were lower in cold acclimated insects. For C. ferrugineus, alanine, aspartic acid, threonine, valine, isoleucine, leucine, phenylalanine and phosphoethanolamine were higher in cold acclimated insects. For both species tyrosine was lower in cold acclimated insects. There were small but significant differences between Canadian and French strains of S. granarius, with the Canadian strain being more cold hardy and having higher levels of trehalose. There were small but significant differences between male and female S. granarius, with males being more cold hardy and having higher levels of proline, asparagine and glutamic acid. In conclusion, high levels of trehalose and proline were correlated with cold tolerance, as seen in several other insects. However, correlation does not prove that these compounds are responsible for cold tolerance, and we outline further tests that could demonstrate a causal relationship between trehalose and proline and cold tolerance.     

Isolation of alcohol tolerant,osmotolerant and thermotolerant yeast strains and improvement of their alcohol tolerance by UV mutagenesis

In this study, the yeast strains were isolated from grapes by serial dilution technique to determine their alcohol-, sugar- and thermotolerance. 34 wild type yeast strains were isolated and alcohol-, sugar- and thermotolerance of these strains were determined. The maximum alcohol tolerance was found to be 9% (v/v) in yeast strain which is named Y2. Thermotolerance behavior of 6 strains were investigated. The strains were treated with UV light with intervals of 20, 30, 40 and 50 seconds. Selected resistant colonies were investigated for alcohol tolerance. It was found that alcohol tolerance increased from 9% (v/v) to 12% (v/v) on Y2 strain.     

桔小实蝇对敌百虫抗性稳定性及再增长趋势

通过室内试验,研究了用LC₅₀及LC₉₀两种浓度敌百虫汰选14代建立的不同程度高抗性水平的桔小实蝇Bactrocera dorsalis抗性品系TrR₅₀和TrR₉₀在停止使用药剂后的抗性衰退规律,以及抗性衰退到原有水平的1/3时再用药汰选的抗性再增长规律。结果表明：两抗性品系对敌百虫抗性均不稳定,但抗性衰退速率不同,TrR₉₀完全隔离药剂4代后,抗性已衰退至原有水平的1/3,而TrR₅₀完全隔离药剂7代后,抗性才衰退至原有水平的1/3,但随后两品系抗性衰退均减缓,到19代后抗性均还处于低水平抗性阶段,无法衰退至敏感水平。通过对其抗性衰退趋势进行方程拟合,结果表明两品系抗性衰退均符合S型曲线模型。抗性再增长试验结果表明：桔小实蝇两抗性品系对敌百虫抗性再增长趋势不同,TrR₅₀继续使用药剂汰选,经过6代选育,抗性迅速上升,接近原有的抗性水平,随后保持平稳增长; 而TrR₉₀继续使用药剂汰选,经过9代选育,抗性才迅速上升,经过12代选育,抗性才接近原有的抗性水平。通过对两品系抗性再增长趋势进行方程拟合,结果表明两品系抗性再增长趋势不同,TrR₅₀品系抗性再增长趋势符合逻辑斯蒂模型,而TrR₉₀品系则符合二次曲线模型。     

Genetic differences in physiology, growth hormone levels and migratory behaviour of Atlantic salmon smolts

Out of five strains of Atlantic salmon Salmo salar of 1+ years released upstream of a fyke net in the River Gudenaa in 1996, three, Lagan, Ätran and Corrib, migrated immediately, 50% of the recaptured fish reaching the net in 3–6 days. Burrishoole and Conon fish migrated with a 15–19 day delay. Smolt development in 1997 at the hatchery showed a spring surge in gill Na⁺, K⁺-ATPase activity in all strains which was correlated with increased seawater tolerance. Differences in the timing of gill enzyme development matched the observed migration pattern well. Lagan, Ätran and Corrib strains reached high enzyme activity earlier than the Burrishoole and Conon strains, and strains with delayed enzyme development and migration showed a delayed regression of seawater tolerance compared with the early strains. Inter-strain differences in plasma growth hormone profiles could not be related to the observed patterns of Na⁺, K⁺-ATPase and seawater tolerance development. The study gives evidence of genetic influence on the timing and intensity of smolting and subsequent migration in Atlantic salmon.     

Isolation of clonal strains of chicken embryo fibroblasts

A method for cloning of chicken embryo fibroblasts (CEF) was developed, yielding a cloning efficiency of up to 50% without use of feeder cells or conditioned medium. An analysis of the growth potential of over 200 randomly selected clones showed that only approx. 4% of the clones were capable of doubling more than 35 times before undergoing cellular senescence. A positive correlation between initial growth rate and in vitro lifespan was observed. This served as a basis for a simple selection procedure for fibroblast strains suitable for long-term culturing. None of over 200 clones thus isolated could be established into a line. Subclones from clonal CEF strains were more homogeneous than uncloned CEF cultures with respect to morphology and growth behaviour, but still heterogeneous in their in vitro life span. All fibroblast strains tested could be effectively infected and transformed by a variety of avian sarcoma and leukosis viruses.     

Selection of high ethanol-yieldingSaccharomyces

The effect of ethanol concentrations in the medium, upon yeast growth was studied. The results obtained showed that different strains differ in their alcohol tolerance, expressed as units of turbidity readings. However, at 12% all strains almost stopped growing. In a trial to obtain more tolerant strains, the training method by successive transfers was used. Nevertheless, no increase in tolerance could be gained.     

Comparative study on the antimicrobial effect of 0.5% chlorhexidine gluconate and 70% isopropyl alcohol on the normal flora of hands.

A gloved-hand wash method was used to compare the antimicrobial effect of chlorhexidine gluconate alcohol emollient hand wash (HIBISTAT) with that of 70% isopropyl alcohol on the normal flora of the hands (81 subjects) under conditions designed to mimic use by surgeons. Results of the immediate postwash effects on the bacterial counts for all 3 tests days showed that chlorhexidine significantly reduced the normal flora of the hands. When compared with the base line bacterial counts, there was 85, 96, and 98% reduction with chlorhexidine treatment and 84, 93, and 90% reduction with alcohol treatment on days 1,2, and 5, respectively. The difference between chlorhexidine and alcohol treatments was not statistically significant on days 1 and 2, but was significant on day 5 (P less than 0.01). For delayed postwash bacterial counts (for persistent antimicrobial effects), the overall log means were 4.9943 and 5.4684 for chlorhexidine and alcohol treatments, respectively. The difference between the two treatments was significant (P less than 0.01). After the chlorhexidien treatment, there was no significant growth of bacteria over a period of 6 h when compared with the base line bacterial counts.     

Comparative study on the antimicrobial effect of 0.5% chlorhexidine gluconate and 70% isopropyl alcohol on the normal flora of hands.

A gloved-hand wash method was used to compare the antimicrobial effect of chlorhexidine gluconate alcohol emollient hand wash (HIBISTAT) with that of 70% isopropyl alcohol on the normal flora of the hands (81 subjects) under conditions designed to mimic use by surgeons. Results of the immediate postwash effects on the bacterial counts for all 3 tests days showed that chlorhexidine significantly reduced the normal flora of the hands. When compared with the base line bacterial counts, there was 85, 96, and 98% reduction with chlorhexidine treatment and 84, 93, and 90% reduction with alcohol treatment on days 1,2, and 5, respectively. The difference between chlorhexidine and alcohol treatments was not statistically significant on days 1 and 2, but was significant on day 5 (P less than 0.01). For delayed postwash bacterial counts (for persistent antimicrobial effects), the overall log means were 4.9943 and 5.4684 for chlorhexidine and alcohol treatments, respectively. The difference between the two treatments was significant (P less than 0.01). After the chlorhexidien treatment, there was no significant growth of bacteria over a period of 6 h when compared with the base line bacterial counts.     

Selection <Emphasis Type="Italic">in vitro</Emphasis> and accumulation of phytochelatins in cadmium tolerant cell line of cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis>)

Cucumber (Cucumis sativus L.) cells from suspension culture were selected for their ability to grow and divide rapidly in toxic concentration of cadmium. As a result of selection a cell suspension tolerant to 100 M cadmium chloride (CdCl₂) was initiated. The selected tolerant line exhibited stable and repeatable increase in fresh and dry weight of cells in the presence of cadmium. The accumulated level of phytochelatins in cadmium sensitive (unselected) and tolerant cell line was measured by high performance liquid chromatography (HPLC) after 3, 24 h and 5 days of cadmium treatment. It was shown that in both cell lines Cd induced accumulation of phytochelatins and simultaneous glutathione depletion occurred. No distinct changes were found after 3 and 24 h of cadmium treatment whereas after 5 days of exposure to the metal, the level of phytochelatins was two times higher in the sensitive cell line as compared to the tolerant one. The accumulation of phytochelatins was correlated with cadmium concentration that increased in both cell lines during the course of cell exposure to metal. However, the level of cadmium was always lower in the tolerant cell line. The results showed no direct correlation between the tolerance of cucumber cells to Cd and the accumulated level of phytochelatins. Other mechanisms responsible for the increased tolerance of cucumber cells exposed to Cd are discussed.     

Salinity tolerances of 62 strains of Pflesteria and Pfiesteria-like heterotrophic flagellates (Dinophyceae)

The salinity tolerance of 62 strains of Pfiesteria and Pfiesteria‐like heterotrophic dinoflagellates was measured. All strains were acclimated at 12 psu for at least 1 year before experimentation. Strains isolated from the Chesapeake Bay and Neuse River systems tolerated lower salinities than strains isolated from the Wilmington River system (P< 0.005). Swimming cells were still observed after 5 days at 0.5 psu for one strain, and at 1 psu for most other Chesapeake Bay and Neuse River strains. Swimming cells for the Wilmington River were still observed after 5 days at 3–5 psu, but no swimming cells were observed at ≤ 2 psu. With regard to the upper salinity tolerance, the Wilmington River strains tolerated higher salinities than the Chesapeake Bay and Neuse River systems (P< 0.005). Most Wilmington River strains were swimming after 5 days at salinities ≥ 50 psu, whereas the Chesapeake Bay and Neuse River system strains rarely had swimming cells at salinities exceeding 35–45 psu. For all three water systems and for both lower and higher salinities, cells apparently encysted in many instances. However, when salinities were returned to 12 psu, swimming cells often re‐appeared. Statistically significant geographic differences in salinity tolerance suggest a geographic adaptation has occurred and that salinity tolerance is under genetic control. The results also suggest there is diversity among the strains.     

ADAPTATION TO FERMENTING RESOURCES IN DROSOPHILA MELANOGASTER: ETHANOL AND ACETIC ACID TOLERANCES SHARE A COMMON GENETIC BASIS

Ethanol and acetic acid tolerances were compared in a French, highly tolerant population, and in a Congolese, very sensitive population. For both tolerances, chromosome substitutions demonstrated a major effect on chromosome 3, a lesser effect on chromosome 2, and no effect on chromosome 1, except in interactions. Directional selection experiments led to significant increases of tolerance to both toxics. Of greater interest, a strong correlated response was observed in each line: increased ethanol tolerance was accompanied by higher acetic acid tolerance and vice versa. A high genetic correlation (average value r = 0.77) was found between the two traits. These data suggest that alcohol dehydrogenase (ADH) activity does not play a major role in explaining the physiological differences known between Afrotropical and European populations. The metabolic flux permitting the detoxification of ethanol and acetic acid seems to be mainly controlled by acetyl-coA synthetase (ACS) at least in adult flies. Acetic acid adaptation could be as important as ethanol adaptation in the ecology of Drosophila melanogaster and other Drosophila species.     

Induction of freezing tolerance in potato (Solanum commersonü) suspension cultured cells

The induction of freezing tolerance by abscisic acid (ABA) or cold treatment in suspension cultured cells of Solanum commersonii was studied. Both ABA (50–100 μ M ) at 23°C and low temperature (4°C) increased freezing tolerance in cultured Solanum commersonii cells from a LT₅₀ (freezing temperature at which 50% cells were killed) of —5°C (control) to —11.5°C in 2 days. Cold-induced freezing tolerance reached its maximum at 2 days and remained constant throughout the cold acclimation period of 11 days. The freezing tolerance induced by ABA, however, showed a rapid decline 2 to 5 days after initiation of ABA treatments. Addition of ABA (100 μ M ) to the culture medium at the inception of low temperature treatment did not enhance freezing tolerance of the cells beyond the level attainable by either treatment singly. Poly(A⁺)-RNA was isolated from the respective treatments, translated in a rabbit reticulocyte lysate cell free system, and the translation products were resolved by two dimensional polyacrylamide gel electrophoresis (ID-PAGE). Analysis of the in vitro translated products revealed changes in the abundance of approximately 26 products (encoding for polypeptides with M, of 14 to 69 kDa and pl of 4.90 to 6.60) in ABA-treated cells 12 h after treatment, and 20 (encoding for polypeptides with M_r of 12 to 69 kDa, with pl of 4.80 to 6.42) in cells exposed to 4°C for 12 h. There were only 5 novel translation products observed when the ABA-treated cells reached the highest level of freezing tolerance (2 days after the initiation of ABA treatment). Changes in translatable RNA populations during the induction of freezing tolerance in cells treated with either ABA or low temperature are discussed.     

Probiotic characteristics of Lactobacillus strains isolated from cheese and their antibacterial properties against gastrointestinal tract pathogens

In the present study, four Lactobacillus strains from the cheese were analyzed for its probiotic potential against enteropathogenic bacteria. The probiotic properties of the selected strains were also analyzed and the selected bacterial strains showed high tolerance in bile salts and organic acid. The strain L. plantarum LP049 showed maximum survival rate (92 ± 4.2% and 93.3 ± 2%) after 3 h of treatment at 0.25% (w/v) bile salts and 0.25% (w/v) organic acid concentrations. The ability of the Lactobacillus strains to adhere to human epithelial cells (HT-29 cell lines) was evaluated and L. plantarum LP049 showed maximum adhesion property (19.2 ± 1.1%) than other tested strains. The Lactobacillus strains produced lactic acid at various concentrations. Compared with other strains, maximum level of lactic acid (3.1 g/L), hydrogen peroxide (4.31 mM) and bacteriocin (31 AU/mg) was detected in LB049. The inhibitory activity of culture supernatant against various bacterial pathogens was observed. The zone of inhibition ranged between 6 ± 2 mm and 23 ± 2 mm. The cell free extract showed activity against, Escherichia coli (ATCC 10536), Salmonella enteritidis (ATCC 13076), Shigella flexneri (ATCC 29903), and Enterococcus faecium (ATCC 8459). Consequently, L. plantarum LP049 may be considered as a potential candidate for the production of novel bioactive metabolites for therapeutic and bio-protective applications.     

The Response of Sugarcane (Saccharum officinarum) Cultured Cells to Anoxia and the Selection of a Tolerant Cell Line

The effect of anoxia on the sugarcane (Saccharum officinarum L.) cultured cells was studied in order to elaborate a technique for in vitro selection of cell lines, which would be tolerant to anaerobic stress. Inhibitory and lethal doses of anaerobic incubation were established from the state of the mitochondrial ultrastructure during the anaerobic incubation of cells either with or without exogenous glucose, as well as from the pattern of the post-anaerobic callus growth. An intact state of the mitochondrial ultrastructure and the viability of some cells in the presence of 3% glucose were shown to be maintained for at least 14 days of anaerobic incubation, while the index of post-anaerobic growth decreased by almost 50% even after 72-hour-long anaerobiosis. In the absence of exogenous glucose, a marked destruction of mitochondria and a twofold decrease in the callus growth index were observed as early as after six-hour-long anaerobic stress. A 48-hour-long incubation under these conditions resulted in the maintenance of the intact ultrastructure only in 7–10% of cells, while a 96-hour-long anaerobiosis brought about the complete degradation of the subcellular structure and cell death. A 48-hour-long anaerobiosis without exogenous glucose was chosen for selecting the anoxia-tolerant cell lines. After three cycles of selection, the anoxia tolerance of the selected cell line exceeded the respective index of the initial callus several-fold. In selected line, about 50% of cells retained viability and could resume growth even after 96-hour-long anaerobic incubation. The experimental results obtained were used to determine the possible causes of the heterogeneity of callus cells as regards their anoxia resistance.     

水稻籼爪重组自交系群体芽期耐旱性鉴定

以20%PEG-6000作渗透介质,模拟干旱条件,芽期处理水稻籼爪重组自交系7天,测定处理后的胚根长、芽长、芽干重。以各性状相对值的耐旱级别而得出的耐旱总级别为指标,筛选出8个耐旱株系,它们可作为耐旱资源在育种和耐旱机理研究中应用。     

Lipase Activity of Mucor pusillus

Two strains of Mucor pusillus were examined for their ability to synthesize lipase in a complex medium used in the production of milk-clotting protease. Lipase activity of both strains reached maximal after 6 days of incubation under submerged conditions at 35 C. Lipase secreted into the medium hydrolyzed butterfat and vegetable lipids, as well as selected synthetic triglycerides. About 50% of lipase activity was destroyed after a 45-min heat treatment at 58 C.     

Morphological and functional changes in the rat heart after X irradiation: strain differences

The hearts of mature male rats of the Wistar and Sprague-Dawley strains were locally irradiated with single doses of 17.5 and 20.0 Gy of X rays, respectively. These two dose levels had previously been shown to result in a comparable latent period between irradiation and the death of rats of these two strains from cardiac failure. Morphological changes in the myocardium and modifications in cardiac function were assessed in the animals at 28, 70, and 100 days after irradiation. The first radiation-induced change which was observed in the myocardium was a rapid decline in capillary density and a loss of alkaline phosphatase activity by the capillary endothelial cells. The capillary density was reduced to approximately 50% of that of unirradiated control values at 28 days and to approximately 40% of the control values between 70 and 100 days after irradiation. The loss of enzyme activity was also detected at 28 days. Examination of histological sections showed an increase by 70 days in the areas with negative enzyme activity up to approximately 70% of the myocardium. The reduction in capillary density and the loss of enzyme activity occurred before any marked pathological changes were seen in the myocardium. The pathological lesions seen in the myocardium at 100 days after irradiation were qualitatively and quantitatively the same in the two strains of rat. Measurements of cardiac output in Sprague-Dawley rats showed a gradual decline in output after irradiation; however, measurements in Wistar rats showed a progressive increase in cardiac output over the same period of time. It was shown by rubidium extraction that there was an increase in the percentage of the total cardiac output distributed to the ventricular muscle of Sprague-Dawley rats, while similar measurements in Wistar rats showed no significant change. In spite of the marked strain differences observed in cardiac output and rubidium extraction, blood perfusion per gram of ventricular muscle was apparently not modified in both strains of rat after irradiation. These findings indicated that the correlation between morphological effects after irradiation and the functional expression of damage is highly complex.     

兴安落叶松游离氨基酸的动态变化与损伤因子的关系

为研究不同损伤因子及其不同损伤程度对落叶松游离氨基酸组分及其含量的影响,用剪叶和落叶松毛虫(Dendrolimussuperans)取食损伤兴安落叶松(Larixgmelinii)幼苗,使用HPLC柱前AccQ.Tag衍生法测定针叶内游离氨基酸的组成及含量的变化。结果表明,松针中含有17种游离氨基酸,以苯丙氨酸含量最高,损伤15d后,剪叶4枝比对照增加36.6%,虫害4枝增加的百分率达到98.3%。剪叶4枝15d后及虫害4枝5d后,损伤25%与50%之间氨基酸总含量差异显著(P0.05),且昆虫取食4枝25%、50%、75%后松针内游离氨基酸总含量的变化趋势与剪叶处理后总氨基酸含量的变化一致,均在处理10d后呈现较高的含量,虽虫害后的氨基酸总含量高于剪叶损伤后的总含量,但二者之间差异不显著。剪叶4枝、8枝、12枝3个损伤程度后针叶内的游离氨基酸含量无显著差异,说明剪叶4枝即能超过落叶松的耐受阈值,达到防御诱导效应。两种处理下多种游离氨基酸的含量均有下降,其中8种氨基酸含量存在显著差异(P0.05),即丝氨酸、谷氨酸、甘氨酸、组氨酸、脯氨酸、半胱氨酸、赖氨酸和苯丙氨酸。表明人工模拟剪叶的损伤方式与虫害都可诱导松树氨基酸组分及含量的变化,而间接影响害虫的生长发育。     

Resistance of selected lines of Haemonchus contortus to thiabendazole, morantel tartrate and levamisole.

A field strain of Haemonchus contortus isolated from sheep at Armidale N.S.W., was found to be resistant to thiabendazole with approximately 20 per cent of the worms surviving a 50 mg/kg dose. The isolate was selected over six generations for resistance to 50 mg/kg thiabendazole. After this time, selection on one line was continued at 50 mg/kg thiabendazole and selection on a second line was extended to include 8·8 mg/kg morantel tartrate. A drug tolerance assay on the third generation of the thiabendazole-morantel tartrate selected line showed the ld_5o to be 5·3 mg/kg and the ld₉₅ to be 18 mg/kg morantel tartrate; the reported ld₅₀ and ld₉₅ for non-resistant strains of H. contortus to morantel tartrate are 2·5 mg/kg and 8 mg/kg respectively. In the fourth generation the thiabendazole and the thiabendazole-morantel tartrate selected lines together with a recently isolated field strain were assayed for resistance to thiabendazole, morantel tartrate and levamisole. The results indicated that the resistance to thiabendazole was probably due to a single gene. Both selected lines were more resistant to thiabendazole than the field strain. The thiabendazole-morantel tartrate selected line was more resistant to morantel tartrate than either of the other two. Resistance to morantel tartrate appeared to be polygenic in nature and due to increased vigour. The lowest dose of levamisole (1·6 mg/kg) killed more than 95 per cent of all strains of worms. There was no significant increase in effectiveness at higher dose rates indicating that surviving worms were resistant to the drug.