Directional and correlated effects of selection on amylase activity,weight and developmental time in Tribolium confusum (Coleoptera,Tenebrionidae)

The interrelationships of amylase activity, weight and developmental time in a strain of Tribolium confusum were studied by selecting, in turn, for one of these traits and monitoring other, non-target characters, in all lines.Experimental lines were selected for high and low amylase activity (AH, AL), for heavy and light adult weight (WH, WL) and for fast and slow development (DF, DS). The variables monitored every generation in all lines were mean amylase activity (measured colorimetrically), mean individual adult weight, median egg-to-pupa developmental time, mean productivity (offspring per fertile pair) and % sterile pairs.The six lines may be grouped by their response to selection in a meaningful way. AH, WH, and DS all had higher amylase activity, heavier adult weight, slower development and lower fitness (lower productivity, higher percent sterility) than the corresponding AL, WL, and DF, although a different variable was under selection in each pair of lines. These results demonstrate that amylase activity, developmental time and individual weight are intercorrelated.     

Screening of marine actinobacteria for amylase enzymes inhibitors

Amylase inhibitor producing actinobacteria were isolated and characterized from terrestrial environment and there is no much report found from marine environment, hence in the present study, 17 strains isolated from the rhizosphere sediments of mangroves were tested for their amylase inhibition ability. Seawater requirement test for the growth of actinobacteria found that the strains SSR-3, SSR-12 and SSR-16 requires at least 50% and SSR-6 requires at least 25% seawater for their growth. The inhibition activity of both prokaryotic and eukaryotic amylase was tested by using Bacillus subtilis and Aspergillus niger. The maximum amylase activity (40mm) produced by the A. niger was taken as positive control, when the test actinobacteria strains grown in the medium they inhibited amylase activity and was evidenced by the reduction in inhibition zone (14–37 mm) similarly the amylase produced by the Bacillus subtilis was also recorded maximum (35 mm) amylase activity and was taken as positive control, and the test atinobacterial strains reduced enzyme action(12–33 mm) it varied levals. This indicates that the actinobacteria strains were controlled amylase enzyme activity in both the cases. The strain SSR-10 was highly effective and SSR-8 was less effective in inhibiting eukaryotic amylase produced by A. niger. The strain SSR-2 was effective and SSR-6 showed very less effect in inhibiting the prokaryotic amylase produced by the B subtilis.     

Adipokinetic hormones control amylase activity in the cockroach (Periplaneta americana) gut

This study examined the biochemical characteristics of α‐amylase and hormonal (adipokinetic hormone: AKH) stimulation of α‐amylase activity in the cockroach (Periplaneta americana) midgut. We applied two AKHs in vivo and in vitro, then measured resultant amylase activity and gene expression, as well as the expression of AKH receptor (AKHR). The results revealed that optimal amylase activity is characterized by the following: pH: 5.7, temperature: 38.4 °C, K_m (Michaelis–Menten constant): 2.54 mg starch/mL, and V_max (maximum reaction velocity): 0.185 μmol maltose/mL/min. In vivo application of AKHs resulted in significant increase of amylase activity: by two‐fold in the gastric caeca and 4–7 fold in the rest of the midgut. In vitro experiments supported results seen in vivo: a 24‐h incubation with the hormones resulted in the increase of amylase activity by 1.4 times in the caeca and 4–9 times in the midgut. Further, gene expression analyses reveal that AKHR is expressed in both the caeca and the rest of the midgut, although expression levels in the former were 23 times higher than levels in the latter. A similar pattern was found for the amylase (AMY) gene. Hormonal treatment did not affect the expression of either gene. This study is the first to provide evidence indicating direct AKH stimulation of digestive enzyme activity in the insect midgut, supported by specific AKHR gene expression in this organ.     

Location of the α-amylase gene in rumenStreptococcus bovis strains distinguished by unstable amylase activity

Genetic stability of amylase activity after serial subcultivation experiments with amylolytic ruminalStreptococcus bovis strains was investigated. Two strains Amy⁺ and Amy⁻ were obtained. Loss of amylase activity connected with the loss of plasmid DNA was not found in these strains. The presence of the gene responible for the amylase activity in the chromosome of these strains was revealed by hybridization of the α-amylase gene on pJK108 against chromosomal DNA ofS. bovis andBacillus subtilis after a complete restriction withEcoRI.     

Functional significance of amylase polymorphism in Drosophila melanogaster. III. Ontogeny of amylase and some α-glucosidases

Changes in amylase (E.C. 3.2.1.1), maltase (E.C. 3.2.1.20), sucrase, and PNPGase activities in relation to changes in wet weight and protein content were studied during the development of larvae and adult flies from two strains of Drosophila melanogaster, homozygous for different amylase alleles. All -glucosidase activities increase exponentially during a large part of larval development, parallel to the increase in weight, and drop at the end of the third instar. Amylase activity of the Amy ¹ strain follows the same pattern. In contrast, amylase activity of the Amy ^4,6 strain continues its exponential increase longer. In the third larval instar amylase activity in the Amy ^4,6 strain becomes much higher than in the Amy ¹ strain. During the first hours of adult life amylase activity of the two strains does not differ. Then Amy ^4,6 activity starts to rise and becomes much higher (4–5 times) than Amy ¹ amylase activity, which remains approximately constant. All adult enzyme activities are much higher than in larvae. Comparison of enzyme activity of amylase and -glucosidases in larvae and adults confirms that differences in amylase activities can become important only when starch is a limiting factor in the food.The investigations were supported by the Foundation for Fundamental Biological Research (BION), which is subsidized by the Netherlands Organization for the Advancement of Pure Research (Z.W.O.).     

Some Biochemical Abnormalities Caused by Cypermethrin in Adults of Tribolium castaneum (Herbst.)

Biochemical effects of sub lethal doses LC₁₀ and LC₂₀ of cypermethrin were studied on some enzymes and macromolecule activities of adult beetles of Tribolium castaneum (Herbst.). Cypermethrin caused disturbances in levels of all biochemical components under study. The dose of 0.78 ppm caused abnormalities in α‐amylase and FAA by increasing their activities i.e., 45.45% and 21.97% significantly. The higher sub lethal dose of 2.62 ppm disturbed all the parameters (AcP, α‐amylase, soluble protein and FAA) except AkP, which was decreased by 93.06%. Moreover, sub lethal doses either increased or decreased the levels of all parameters non‐significantly except AkP and FAA which were effected significantly by 87.92% and 14.29% at lower and higher doses, respectively. In the present studies, cypermethrin significantly enhanced the activity of AkP in both susceptible and resistant strains of T. castaneum adult beetles while FAA contents were increased significantly in resistant strain only. The activity of α‐amylase was significantly lowered in susceptible strain only.     

Production of Industrial Enzymes (Amylase,Carboxymethylcellulase and Protease) by Bacteria Isolated from Marine Sedentary Organisms

The abilities of bacteria isolated from eight marine sedentary organisms, six marine sponges (Spirastrella sp., Phyllospongia sp., Ircinia sp., Aaptos sp., Azorica sp. and Axinella sp.), one soft coral (Lobophytum sp.) and one alga (Sargassum sp.) to produce industrial enzymes (amylase, carboxymethylcellulase and protease) were examined. The mean total viable counts of the bacterial isolates ranged from 8.7 × 10⁴ to 8.4 × 10⁵ cfu/g wet weight of the organism. All eight organisms harboured amylase (0.05–0.5 IU/ml), carboxymethylcellulase (0.05–0.5 IU/ml) and protease (0.1–0.5 IU/ml) producing bacteria. Of 56 bacterial strains tested, as many as 60 to 83% of the strains produced at least one of the three enzymes, and 47% of strains were able to produce all three enzymes. High activities (> 0.5 IU/ml) of the three enzymes were recorded in bacterial strains belonging to the genera Alcaligenes and Bacillus. From the results of this study, it appears that bacteria associated with marine sedentary organisms are the novel source of industrial enzymes for possible commercial applications and may play an important role in enzyme‐catalysed organic matter cycling in marine environments.     

Variation between field and laboratory populations of Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae)

Rates of development and pupal weights were compared between seven field populations and two laboratory strains of Tribolium castaneum. Rates of development did not differ significantly between field populations, but pupal weights did vary. Laboratory strains were both slower to develop and heavier than the field populations. The data support the hypothesis that field populations undergo selection for fast development in transient habitats. The findings are discussed in the context of r and K selection in field populations and laboratory cultures.     

The effects of copper, iron and zinc on digestive enzyme activity in the hybrid tilapia Oreochromis niloticus (L.) ×Oreochromis aureus (Steindachner)

The present experiment was conducted to study effects of Cu, Fe and Zn on activities of digestive enzymes of the hybrid tilapia Oreochromis niloticus×Oreochromis aureus. The acidic protease activities increased 65·5 and 55·1% by addition of homogenates of digesta‐containing stomach with copper (75 mg l⁻¹) and zinc (50 mg l⁻¹) respectively. Addition of Cu and Zn increased the activities of protease in the hepatopancreas homogenates by 132·7 and 38·1% respectively, and reduced the activity of protease in the digesta‐containing intestine homogenates by 11·0 and 13·8% respectively. Addition of Fe (50 mg l⁻¹) increased the acidic protease activity by 96·7% but did not alter the activities of protease in the intestine and hepatopancreas. Addition of Cu markedly inhibited activities of amylase in intestine and hepatopancreas homogenates, while Zn addition showed no effects. Addition of Fe reduced activities of amylase in the intestine homogenates by 47·9% but had no effect on amylase activities in the hepatopancreas. When Cu (75 mg kg⁻¹), Fe (50 mg kg⁻¹) and Zn (50 mg kg⁻¹) were supplemented to basal diet for 3 weeks, the activities of amylase in hepatopancreas homogenates increased 125·3, 215·6 and 70·0%, respectively, the activities of amylase in intestine increased 79·8, 74·6 and 48·5%, respectively, and the activities of lipase in intestine increased 90·5, 149·8 and 84·0%, respectively. Supplementation of Cu, Fe or Zn into diet had no effects on activity of protease in all digestive organs. Therefore, the results suggest that effects of Cu, Fe and Zn on activity of digestive enzymes in vitro were different from those seen in vivo, and that the positive effects of Cu, Fe and Zn supplemented to fish diet would be valuable information for formulating fish feed.     

发酵车间空气源细菌淀粉酶产生菌筛选及多样性分析

采用平板水解圈法(plate hydrolysis spot method)对分离自酒鬼酒发酵车间空气样品的细菌进行淀粉酶产生菌筛选,运用基于16S rRNA基因序列的分析方法对高酶活菌株进行系统发育多样性分析。结果表明,73个受试菌株中,有23株为淀粉酶产生菌,占受试菌株的31.5%,其中有9株为高酶活菌。23个淀粉酶产生菌类群多样性和物种多样性较高,属于4个大的系统发育类群(Actinobacteria、Deinococcus-Thermus、Firmicutes、Proteobacteria)中的10个科(Bacillaceae、Deinococcaceae、Intrasporangiaceae、Microbacteriaceae、Micrococcaceae、Nocardiaceae、Propionibacteriaceae、Pseudomonadaceae、Rhodobacteraceae、Xanthomonadaceae)的13个属,可分为21个物种。进一步分析表明,9株高酶活菌属于细菌域(Eubacteria)的4个大的系统发育类群(Actinobacteria、Deinococcus Thermus、Firmicutes、Proteobacteria)的7个科(Bacillaceae、Deinococcaceae、Micrococcaceae、Microbacteriaceae、Nocardiaceae、Rhodobacteraceae、Xanthomonadaceae),归属于8个属。研究结果表明,酒鬼酒发酵车间空气源细菌存在较高比例的淀粉酶产生菌,且这些菌株具有较高的类群多样性和物种多样性。     

Isolation of quizalofop-resistant mutants of Nannochloropsis oculata (Eustigmatophyceae) with high eicosapentaenoic acid following N-methyl-N-nitrosourea-induced random mutagenesis

Nannochloropsis oculata was subjected to N-methyl-N-nitrosourea-induced mutagenesis under the selection pressure of quizalofop, a known inhibitor of acetyl-CoA carboxylase (ACCase) activity with the objective of generating genetically tractable mutants with altered fatty acid metabolism. Two mutants, QUIZ1 and QUIZ2, with stable resistance to quizalofop were isolated and partially characterized. The growth properties and morphology of the mutants appeared identical with the parent strain. However thermo-tolerance was observed in the mutants. Enhanced resistance to quizalofop suggested the presence of herbicide resistant isoforms of ACCase. In vitro assays for ACCase activity showed that ACCase in the wild strains was much more sensitive to quizalofop than the mutant strains. Gas chromatographic analysis of fatty acids revealed that the mutant strains were rich in polyunsaturated fatty acids (n– 3PUFAs), as well as total fatty acid contents; this was accompanied by a concomitant increase in triacylglycerol (TAG) followed by linoleic acid (18:2), arachidonic acid (20:4 n– 6) and EPA (20:5 n– 3). These results suggest that an increased substrate pool (malonyl-CoA) (due to increased specific activity of ACCase) in the mutant strains in vivo and in vitro may have led to the increased TAG accumulation. Random mutagenesis was shown to be a good tool to manipulate PUFAs and EPA in Nannochloropsis. The strains developed will be useful in understanding fatty acid metabolism using genetic and biochemical approaches and also for their direct use in mariculture.     

Phenotypic diversity and technological properties of <Emphasis Type="Italic">Bacillus subtilis</Emphasis> species isolated from <Emphasis Type="Italic">okpehe</Emphasis>, a traditional fermented condiment

The Bacillus subtilis wild strains isolated from okpehe, a traditional fermented condiment used as seasoning in Nigeria, the reference and typed strains were investigated for their phenotypic diversity and their technological parameters with a view to obtain adequate data that would enable selection of appropriated starter cultures for vegetable protein fermentation in West Africa. All the 7 strains studied demonstrated diverse phenotypic characteristics and they were identified as Bacillus subtilis, based on the API 50 CHB combined with API 20E profile. Specific sugars that indicated a good hydrolytic potential of the wild strains were fermented. The highest proteinase activity of 90 AU/ml determined quantitatively was observed in the strain Bacillus subtilis BFE 5372, the proteinase was identified by the APIZYM gallery as chymotrypsin. Highest amylase activity of 13 AU/ml was noticed in strain Bacillus subtilis DSM 347 while only 4 strains produced polyglutamic acid with the strain Bacillus subtilis BFE 5359 producing the highest polyglutamate activity of 2.5 mm. Although strain Bacillus subtilis BFE 5301 did not release detectable polyglutamate, the strain demonstrated antagonism against different bacteria and the antimicrobial substance produced by strain Bacillus subtilis BFE 5301 was confirmed as a bacteriocin since its activities were lost after treatment with chymotrypsin and pepsin. The data generated showed the technological parameters that can aid selection of wild strains such as Bacillus subtilis BFE 5301, BFE 5359 and BFE 5372 for optimization of condiment production.     

Comparative genetics of hamster amylases

Syrian (Mesocricetus auratus) and Chinese (Cricetulus griseus) hamsters were phenotyped by electrophoresis for salivary and pancreatic amylases. Syrian hamsters possess two salivary amylase electromorphs, the more anodal (fast) being invariant in 250 outbred and 17 representatives of 5 highly inbred lines. The slow electromorph had activity equal to that of the fast amylase (heavy), or had distinctly less activity (light), or was absent (null). The slow electromorph is inherited as an autosomal semidominant trait with two alleles, Amy ^s and Amy ^o . Amy ^s homozygotes produce heavy, Amy ^o homozygotes null, and heterozygotes light phenotypes, respectively. Five inbred strains of hamsters were homozygous Amy ^o . Pancreatic amylase was monomorphic. Eight outbred Chinese hamsters showed no salivary amylase activity with electrophoresis, but slight activity with long incubation on starch-agar plates. However, pancreatic amylase activity in the Chinese hamster exceeded that in Syrian hamsters. Site duplication and apparent null alleles for amylase genes occur in muroid rodents. The evolutionary implications are discussed.George H. Bunch Chair Professor of Biology     

Improvement of heat and desiccation tolerance in <Emphasis Type="Italic">Heterorhabditis bacteriophora</Emphasis> through cross-breeding of tolerant strains and successive genetic selection

Genetic selection can be a powerful tool to increase beneficial traits in biological control agents. In this study the heat and desiccation tolerance of the entomopathogenic nematode Heterorhabditis bacteriophora Poinar (Rhabditidomorpha: Strongyloidea) were significantly increased by cross breeding tolerant parental strains and successive genetic selection. These strains originated from a prior screening among 60 strains for increased stress tolerance. During genetic selection, the selection pressure was constantly increased and only the most tolerant 10% of the nematode populations were propagated for further selection steps. Assessment of tolerance and selection for both traits was performed with and without prior adaptation to the stress conditions. Eleven selection steps were performed to increase heat tolerance. A final overall increase in mean heat tolerance of 5.5°C was achieved when nematodes had been adapted to heat stress. For non-adapted tolerance an increase of 3.0°C from 40.1°C to 43.1°C was recorded. For comparison, a commercial strain had a mean tolerated temperature after adaptation of 38.2°C and of 36.5°C without adaptation. For assessment of the desiccation tolerance the mean tolerated water activity (a_w-value) of a population was measured. Cross-breeding most tolerant strains reduced the a_w-value from 0.67 to 0.65 after adaptation and from 0.9 to 0.7 without prior adaptation. The following six selection steps could not increase the tolerance whether nematodes had been adapted to stress or not. In comparison, the commercial strain tolerated a mean a_w-value of 0.985 after adaptation and 0.951 without adaptation. Further investigation will have to assess trait stability and possible trade-off effects. This study is a first important step on the road towards domestication of the entomopathogenic nematode H. bacteriophora.     

Control of Rhizoctonia solani and Cotton Seedling Disease by Laetisaria arvalis and a Mycophagous Insect Proisotoma minuta (Collembola)

Population changes in Pseudocercosporella herpotrichoides with and without fungicide selection A conidium mixture of W (wheat-type) and R (rye-type) strains of Pseudocercosporella herpotrichoides (W:R = 1:1) was incubated on PD-agar without fungicide or amended with the imidazole prochloraz or the triazole cyproconazole. The W:R ratio was assessed over four generations using the different benomyl sensitivity of the strains as marker. In untreated mixtures, the R strain completely dominated the population already after two generations. Similarly, in cyproconazole-treated mixtures, the R strain increased to 100% after the second generation, whereas in prochloraz-treated mixtures the R strain either followed the same pattern as in untreated mixtures or fluctuated around the initial proportion of 50% over four generations. The W and R strains displayed different fitness properties. The average hyphal growth rate was 8.5 mm/10 d and 4.5 mm/10 d for the W and the R strains, respectively; the sporulation capacity was 80 and 400 conidia/colony, respectively, for the two strains. The different fitness resulted in a strong dominance of the R strains both in vitro and in field populations towards the end of the vegetation period. This shift towards R-dominance occurred about equally fast with or without fungicide selection.     

Selection of Rhizobium leguminosarum strains for lentil (Lens culinaris) under growth room and field conditions

Most of the production of lentil (Lens culinaris) on the Great Plains occurs on soils that are free of indigenous Rhizobium leguminosarum. Inoculation is required to increase yields through N₂ fixation. A screening program to evaluate the effectiveness of R. leguminosarum strains for lentil was initially carried out under controlled environments followed by an evaluation under field conditions. In two separate growth room experiments, the effectiveness of 185 and 24 different strains of R. leguminosarum were tested for Laird and Eston lentil. Significant differences between strains in number of nodules, shoot weight and nitrogenase activity (acetylene reduction activity, ARA) were found for lentil grown for 5 weeks. When lentil were grown for 7 weeks, significant differences between strains in number of nodules, total plant weight, total N, and % N were observed.Fourteen strains plus Nitragin C inoculant were selected for further field testing on Eston and Laird lentil at two locations in 1986 and one site in 1987. Inoculation increased yield up to 135%. Percent Ndfa and total N₂ fixed ranged from 0 to 76 and 0 to 105 kg ha^-1, respectively. N₂-fixing activity was site specific and higher spring soil NO₃-levels resulted in lower N₂-fixing activity. Depending on site and growing conditions, strains 99A1 and I-ICAR-SYR-Le20 appeared to be superior to the other strains tested. A good agreement was found between the estimates for N₂ fixation based upon the ¹⁵N-isotope dilution and the classical N difference methods. Number of nodules, dry weight of nodules and ARA of Eston and Laird lentil grown under growth room conditions failed to show positive correlations with total dry matter production, total N or total N₂ fixed of field grown lentil. However, total plant weight and total N of lentil grown under growth room conditions were highly correlated with field parameters, and were the most reliable screening parameters for the selection of superior rhizobial strains.     

Identification of intracellular amylase activity in Streptococcus bovis and Streptococcus salivarius

The ruminal bacterium Streptococcus bovis has been demonstrated to produce an extracellular amylase activity. We previously reported on the cloning of a gene from S. bovis encoding for what was initially believed to be the extracellular amylase. DNA sequence analyses indicated that the amylase produced by the cloned gene did not match the N-terminus amino acid sequence of the purified extracellular amylase and contained no apparent leader sequence for secretion. Analyses of crude extracts demonstrated the presence of an intracellular amylase in S. bovis JB1 that differed in molecular weight (56,000) from that of the extracellular amylase (70,000). The 56,000 molecular weight amylase was identical to the amylase produced by Escherichia coli containing the cloned amylase gene. Low levels of intracellular amylase activity were also detected in other strains of S. bovis and also Streptococcus salivarius. Introduction of the plasmid pVA838 containing the cloned amylase gene into S. bovis and S. sanguis resulted in enhanced intracellular amylase production by both organisms. The amylase gene has been sequenced, and analysis of the deduced amino acid sequence for the amylase indicates a high degree of similarity with secreted amylases from Bacillus species.Names are necessary to report factually on available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by USDA implies no approval of the product to the exclusion of others that may also be suitable.     

Isolation and characterization of Paracoccus denitrificans mutants with increased conjugation frequencies and pleiotropic loss of a (nGATCn) DNA-modifying property

A selection scheme was devised to isolate Paracoccus denitrificans mutants with increased recipient qualities in transfer experiments, using broad host range plasmids. In some of the mutants obtained, a DNA modifying activity that prevents the activity of the restriction endonucleases BamHI and BglII on isolated P. denitrificans DNA had simultaneously been lost. From a detailed analysis of the restriction properties of the enzymes SAU3 AI, MboI and DpnI, it was concluded that a subset of GATC sequences in P. denitrificans DNA may be methylated at an unusual position. It was concluded that P. denitrificans possesses at least one potent host-dependent restriction/modification system which affects conjugation. In addition to the class of restriction-defective mutants, at least one other class of enhanced transfer mutants with unknown defect(s) was isolated. Strains, in which the two mutant classes were combined, exhibited transfer frequencies which were significantly higher than strains containing either mutation alone. Such double mutant strains appeared to be well suited for future experiments like complementation analysis, transposon mutagenesis and gene replacement by homologous recombination.     

Relationships between plasma composition and parotid salivary composition and secretion rates in the potoroine marsupials,Aepyprymnus rufescens and Potorous tridactylus

Summary Parotid salivation was investigated in two species of potoroine marsupial, Aepyprymnus rufescens and Potorous tridactylus to ascertain flow rates and composition, the buffer capacity of the saliva with respect to possible dependence of these animals on foregut fermentation, and the similarity of anion excretion patterns to those of the kangaroo parotid. Under anaesthesia neither species secreted spontaneously and secretion was stimulated by intravenous infusion of carbachol, bethanechol and isoprenaline. Under cholinergic stimulation in Aepyprymnus, the concentrations of Na, Cl, HCO₃ and osmolality were positively correlated with flow rate, whereas K, Mg, PO₄, H⁺ and urea were negatively correlated with flow. Amylase activity and the concentrations of protein and Ca showed no consistent relation to flow. Relative to Aepyprymnus, saliva of Potorous had much lower amylase activity and amylase activity per gram protein, lower concentrations of urea and Ca, and higher Na. Protein, K and HCO₃ concentrations were similar in both species. The plasma of both species had similar electrolyte concentrations, but Potorous had lower protein, urea, osmolality and amylase activity. Plasma amylase activity in Aepyprymnus rose during cholinergic stimulation to levels in excess of rodent plasma. Isoprenaline infusion in Aepyprymnus increased salivary amylase activity and concentrations of protein, Ca, HCO₃ and PO₄, and reduced the concentrations of Cl and H⁺. The patterns of anion excretion in the two potoroine marsupials were dissimilar to those of the kangaroo parotid suggesting that parotid fluid secretion is not HCO₃ driven to the same extent as that of kangaroos. Buffer anion concentrations and secretion rates were similar to koalas and low relative to kangaroos, indicating that these potoroines do not rely on foregut fermentation.Abbreviations bw body weight - SEM standard error of mean - VFA volatile fatty acids     

Hybridisation and genetic selection for improving desiccation tolerance of the entomopathogenic nematode Steinernema feltiae

Shelf life of biological control products based on the entomopathogenic nematode Steinernema feltiae is rather limited. In order to prolong shelf life, the metabolism of nematodes during storage must be reduced, either by low temperature or by means of desiccation of the third stage dauer juveniles (DJs). Tolerance to desiccation is limited in S. feltiae. Their tolerance can be increased by an adaptation to moderate desiccation conditions. The objective of this study was to screen for tolerant strains among wild type populations of S. feltiae, hybridise most tolerant strains and further improve desiccation tolerance by subjection of the hybrid strain to genetic selection. Dehydrating conditions, measured as water activity (a _w-values), were produced by treating DJs with different concentrations of the polymer polyethylene glycol 600. Significant variation was recorded among 24 S. feltiae strains. The mean tolerated water activity survived by 50% of the population (WA₅₀) ranged from 0.78 to 0.93 when nematodes were not adapted to desiccation stress and from 0.66 to 0.88 when preadapted to desiccation stress. The six most desiccation tolerant strains of non-adapted and adapted nematode populations were crossed. Preadapted tolerance of hybrids was superior to the tolerance of parental strains, whereas non-adapted tolerance was not increased. The most tolerant hybrid had a WA₅₀ when preadapted of 0.67 and 0.86 when not adapted. The tolerance was lost after few reproductive cycles in the insect Galleria mellonella but was recovered again after six selection cycles with exposure to increasing stress conditions. Virulence and reproduction potential was not negatively affected by the selection. Stabilisation of the selection progress will be a major challenge to enable commercial exploitation of the genetic improvement.