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1.
Injections into 4th instar larvae of Galleria mellonella using either in vitro or in vivo inoculum of the BR-6 isolate of Spiroplasma citri, propagated for one to nine passages, caused 5.7 to 24.7% mortality. Weight gain of the larvae injected at their 4th, 5th, and 6th instar was reduced in the first 4 days after inoculation but final pupal weight of the survivors was not significantly affected. Fourth instar larvae pupated within 10 days after the injection, but more larvae (6–13%) injected with 5th- to 9th-passage cultures pupated 5 or more days later than did larvae injected with 1st to 4th-passage cultures (0–3%). As many as one-third of the injected larvae developed into deformed pupae, with some external appendages missing or with a reduced and distorted thorax or abdomen with uneven tanning of the integument. Spiroplasma multiplied to dense concentrations (108 to 109/ml) in hemolymph smears from injected larvae incubated under oil. Larvae of Tenebrio molitor were not susceptible to S. citri by injection or feeding and G. mellonella were not susceptible by feeding. Transmissibility of S. citri by leafhopper vector to celery and periwinkle plants was retained after propagation for nine successive passages during 7 months in a nonhost insect such as Galleria.  相似文献   

2.
In laboratory experiments with Culicinomyces, the time of death of infected Anopheles hilli larvae decreased as the concentration of conidia to which they were exposed increased. At a concentration of 103 conidia/ml, most first instar and third instar larvae died later than 2 days after exposure and the majority of dead specimens developed external sporulation on the exterior cuticle. On the other hand, at a concentration of 105 conidia/ml, more than 60% of first and third instar larvae died within 2 days of exposure. Very few of these larvae developed external sporulation and it appears that most of them died after penetration by the fungus through the gut cuticle but before the hemocoel was colonized by mycelium. The reason for this rapid death at high concentrations of inoculum is not known, but it is thought that it may be caused by toxic substances associated with the invading hyphae which only attain a lethal titer when massive invasion originates from large numbers of conidia.  相似文献   

3.
Xenorhabdus nematophilus is a pathogenic bacterium causing insect haemolymph septicemia, which leads to host insect death. To address the fundamental mechanisms underlying this haemolymph septicemia, or the immunodepressive response of the host insects following bacterial infection, we tested a hypothesis that the insect immune-mediating eicosanoid pathway is blocked by inhibitory action of the bacterium. Haemocoelic injection of the bacteria into the fifth instar larvae of Spodoptera exigua reduced the total number of living haemocytes with postinjection time and resulted in host death in 16 h at 25 degrees C. The lethal efficacy, described by the median lethal bacterial dose (LD(50)), was estimated as 33 colony-forming units per fifth instar larva of S. exigua. The lethal effect of the bacteria on the infected larvae decreased significantly with the addition of exogenous arachidonic acid (10 μg), a precursor of eicosanoids. In comparison, injections of dexamethasone (10 μg), a specific inhibitor of phospholipase A(2), and other eicosanoid biosynthesis inhibitors elevated significantly the bacterial pathogenicity. Live X. nematophilus induced the infected larvae to form less nodules than did the heat-killed bacteria, but the addition of arachidonic acid increased the number of nodules formed significantly in response to live bacterial injection. The treatment with dexamethasone and other inhibitors, however, decreased the nodule formation after injection of heat-killed bacteria. These results indicate that eicosanoids play a role in the immune response of S. exigua, and suggest strongly that X. nematophilus inhibits its eicosanoid pathway, which then results in immunodepressive haemolymph septicemia.  相似文献   

4.
Soil samples were collected from different rice fields of Singur, Hooghly, West Bengal, India. Spore forming bacteria were isolated from the soil samples and among them, two isolates (BUSNC25 and BUSNC26) were larvicidal against third, fourth and fifth instar larvae of rice leaf folder, Cnaphalocrocis medinalis. The phenotypic, biochemical characterization and 16S rDNA analysis of the two isolates were done. On the basis of phenotypic, biochemical and phylogenetic analysis, the selected bacterial isolates (BUSNC25 and BUSNC26) were identified as Bacillus thuringiensis. The antibiotic sensitivity tests of these two isolates against selected doses of some standard antibiotics were done. Against the 3rd, 4th and 5th instar larvae of C. medinalis, the LC50 values of BUSNC25 were 2.45 × 104, 1.325 × 104 and 2.35 × 104 cfu/ml and of BUSNC26 were 3.375 × 104, 1.9 × 104 and 3.325 × 104 cfu/ml, respectively.  相似文献   

5.
The maximum growth rate occurs in the first half of the 7th and 8th larval instars of the house cricket, at which time food and water consumption is maximal. Growth ceases in the last 2 to 3 days of each instar when food consumption is almost nil. The metabolic rate is twice, and the locomotory activity is four times, higher in the first 2 to 3 days than in the last 2 to 3 days of each instar. The %-gain in dry wt is 120% for the 7th and 139% for the 8th instar. The average digestive coefficient (AD) for the 8th instar is 67% and the average efficiency of food conversion to tissue (dry wt ECI) is 27%. The average daily food consumption for 8th instar larvae is 31.4 mg, and, therefore, we calculate that an average of 12 mg food is burnt per day for energy which is confirmed by the observed average VO2 of 1.2 ml O2/g-cricket/hr. Total lipids as %-total wt (239% mg-gain) increases in the first 4 days then remains constant in spite of cessation of feeding in the last 2 days. The RQ confirms the conversion of carbohydrates to lipids in the first half of each instar. In the moulting cycle carbohydrates are used for maintenance (when not feeding), but lipids are used at apolysis.  相似文献   

6.
《Insect Biochemistry》1989,19(3):323-326
High resolution 31P nuclear magnetic resonance spectroscopy (NMR) was successfully applied to 5th instar larvae of Manduca sexta. Conditions for in vivo analysis under non-saturating conditions are described. The 31P NMR spectrum of intact larvae was composed of six peaks. Their resonance frequencies are reported relative to orthophosphoric acid. Analysis of tissue extracts demonstrated the in vivo peaks to be composed of the β phosphorus resonance of nucleotide triphosphates (NTP) at −19.36 ppm; α phosphorus of NTP and nucleotide diphosphates (NDP) at −10.51 ppm; β and γ phosphorus of NDP and NTP, respectively, at −5.42 ppm; phosphoarginine (PA) at −3.45 ppm; inorganic phosphate (Pi) at +2.76 ppm and sugar phosphates at +3.34 ppm. The major sugar phosphate present in fat body extracts was trehalose-6-phosphate and this was the major phosphorus component of the spectrum of hemolymph. The spin-lattice relaxation times for each in vivo peak were determined.Titration of aqueous fat body and hemolymph extracts was carried out and the relationship between the chemical shift of Pi and pH determined. On this basis the pH of the hemolymph was estimated at approx. 6.7.The metabolic inhibitors, iodoacetate and dinitrophenol, had significant effects on the 31P NMR spectrum of intact larvae. Administration of iodoacetate caused a rapid increase in the levels of sugar phosphates together with decreases in NTP and PA. Dinitrophenol also caused declines in the relative levels of NTP and PA but sugar phosphates decreased as well. The experiments demonstrated the potential of in vivo NMR analysis for metabolic studies on high energy phosphate metabolites in M. sexta.  相似文献   

7.
Continuous flow, laboratory microcosms were used to measure the effects of pH, phenol, and NaCl on the survival of the life stages of Chironomus attenuatus, the caloric content of third and fourth instar larvae and adults, the lipid and protein-nitrogen content of fourth instar larvae, and the interaction among the various responses. The metabolism of phenol was studied using uniformly labeled phenol-C14. pH had a significant effect on the survival of all life stages of C. attenuatus. Survival was higher for all larval instars at pH 7.2 while adult emergence was higher at pH 6.2. Increasing phenol levels resulted in a nearly linear increase in caloric content. Sodium chloride affected the lipid content of fourth instar larvae. The lipid content was higher with NaCl present in the media than without NaCl. Interaction had a significant effect on all responses except survival of third and fourth instar larvae. Phenol-C14 was metabolized by bacteria, but not by C. attenuatus.  相似文献   

8.
The susceptibility of Heliothis armiger larvae of different ages to a commercial nuclear polyhedrosis virus (NPV), Elcar, was determined by bioassay. The median lethal dosage (LD50) increased 150-fold during the first week of larval life at 25°C, i.e., during development to early fourth instar, but daily feeding rate and thus potential virus acquisition also increased. A linear relationship was determined between log LD50 and larval length, indicating that larval length constitutes a useful index for estimating the susceptibility of larval populations. Median lethal times (LT50s) were similar for larvae tested at ages of 0 to 7 days and ranged from 3.6 to 8.0 days at 30°C. The amount of virus produced in a single, infected neonate was equivalent to 1.4 × 106 LD50s for neonates, a 900,000-fold increase on the dose supplied. The data support the practice of directing the NPV against neonates, but, on the basis of larval susceptibility alone, the age of larvae at treatment may not always be critical.  相似文献   

9.
Adoxophyes orana granulovirus (AdorGV) was isolated from overwintering larvae in an orchard in Kent, in the UK. The developmental time of each A. orana instar was determined by measuring the size of the head capsule. The susceptibility of the larvae to the English isolate of AdorGV was evaluated in laboratory bioassays using inoculation by microdroplet feeding and applied dose assays. A series of bioassays were performed to determine LD(50) and ST(50) values for first, fourth and fifth instar larvae. The median lethal doses ranged from 30 occlusion bodies in first instar to 1.36 x 10(6) in fifth instar. The median survival time decreased the later the larvae were infected and ranged from 37 days in first instar to 24 days in fifth instar. Approximately half of the infected larvae released a discharge rich in occlusion bodies from their posterior end prior to death. Approximately 85% of larvae attempted pupation and died as larva-pupa intermediates.  相似文献   

10.
Growth and glycogen content of Chironomus anthracinus in Lake Esrom, Denmark was examined during summer stratification in 1992 and 1993. Simultaneously, effects of oxygen deficiency on glycogen utilization and survival were experimentally studied. The population consisted of almost fullgrown 4th instar larvae in 1992 and 2nd and 3rd instar larvae in 1993. Growth rate and glycogen content changed as hypolimnetic oxygen deficiency increased. During a 1st phase of stratification dry weight and glycogen content increased (2nd and 3rd instars) or was almost constant (4th instar) but decreased significantly during the following 2nd phase. This change from growth to degrowth and utilization of endogenous glycogen reserves correlated with a change in the thickness of the microxic layer (<0.2 mg O2 1–1) above the sediment surface. The layer increased from 2–3 m in phase 1 to 4–5 m in phase 2, and we suggest that this deteriorated the oxygen conditions and resulted in a change in larval energy metabolism from fully aerobic during the 1st phase to partly anaerobic in the 2nd phase. During the 2nd phase larval metabolism was estimated at less than 20% of normoxic rate. Experimental exposure of the larvae to anoxia indicated highly different survival of young larvae (2nd and 3rd instars) and older larvae (large 4th instars). The morality of young larvae was 50% after three days in anoxia at 10 °C, whereas only 25% of the older larvae had died after 3–4 weeks under similar conditions. Extending the treatment, however, resulted in increased death rate of the 4th instar larvae with only 10% surviving after seven weeks. The anaerobic metabolism of 4th instar larvae as estimated from glycogen degradation at 10 °C was 5% of normoxia in the interval from 0–5 days but 1.5% in the interval from 20–25 days. It is concluded that survival of C. anthracinus in anoxia is very limited, but traces of oxygen in the environment allowing for faint aerobic metabolism prolong the survival time of the larvae from a few days (2nd and 3rd instars) or a few weeks (4th instar) to probably 3–4 months.  相似文献   

11.
The insect neuropeptide, [Arg7]-corazonin was injected into larvae of the silkworm, Bombyx mori to investigate its influence on development and behavior. A single injection of 50 pmol of corazonin into the fourth and fifth instar larvae induced prolongation of the spinning period in all experimental groups except for those injected on day 10 of the fifth instar. The injection also caused a prolongation of the pupal period in some experimental groups, while it had no effect on the timing of larval ecdysis and the length of feeding period of the fifth instar. The spinning period was significantly prolonged even at a low dose of 1 pmol. Both the spinning rate and the rate of increase in hemolymph ecdysteroid level during the spinning stage were reduced by injection of corazonin. However, corazonin injection during days 5-7 of the fifth instar reduced the spinning rate without influencing the ecdysteroid level until the end of day 8, thereafter the rate of increase in hemolymph ecdysteroid level was slower in the corazonin-injected larvae than in the control larvae. Therefore, the suppressed ecdysteroid level observed in the corazonin-injected larvae appears to be a result rather than a cause of the reduced spinning rate. This study is the first published report for the corazonin effect on the behavior in insects.  相似文献   

12.
Hemolymph composition of fourth instar larvae of an autogenous strain of Culex pipiens was examined to determine the effects of parasitism by a mermithid nematode, Romanomermis culicivorax. Mosquitoes were reared under two different pH regimens: 4.5 and 7.3. Wet and dry weight of infected mosquitoes reared at either pH were significantly lower than controls. The effects of parasitism in the development of C. pipiens were evaluated from paraffin sections of mosquito larvae 2, 4, and 6 days postinfection. At 2 days postinfection, the infected larvae showed no apparent effects of parasitism; at day 4, the fat body tissue was reduced and imaginal disc development was retarded; and at day 6, parasitized mosquitoes were smaller in cross section, fat body tissue was found only in isolated clumps, and there was a complete absence of imaginal discs. Concentrations of total carbohydrates in hemolymph from infected fourth instar mosquitoes reared at pH 7.3 were reduced. Trehalose and glucose were each reduced by more than half. Total α-amino nitrogen was significantly lower in infected mosquitoes reared at pH 7.3. However, total amino acid concentrations for hemolymph from control and infected larvae reared at pH 7.3 were the same. Methionine sulfoxide decreased 63% and proline increased 2.5 times in infected mosquitoes. Hemolymph protein concentrations were reduced 80% in infected mosquitoes reared at both pHs. The number of hemolymph proteins also declined from 35 to 22 during infection. Two host proteins, 82,000 and 158,000 daltons, remained prominent throughout the mermithid infection.  相似文献   

13.
《Journal of insect physiology》1986,32(12):1065-1073
Topical application of juvenile hormone analogue, methoprene, induced a supernumerary larval moult in the silkworm, Bombyx mori. The incidence changed greatly depending on developmental stages and physiological states of the methoprene-treated larvae. When methoprene was applied to feeding larvae, only those treatments from the middle of the 2nd instar until the middle of the 4th instar were effective. An 18-h starvation period from the beginning of the 4th instar and a dose of 1 μg of methoprene per larva were required for 100% incidence of the perfect superlarvae. Allatectomy had no effects on the induction of superlarvae by methoprene. The treated 4th-instar larvae ecdysed to the 5th instar without any delay compared to the controls, and underwent an additional larval ecdysis 4.5 days later. The induced 6th-instar larvae took 8.5 days until the onset of cocoon spinning. The induced superlarvae showed reduced growth rates but an increase of final mass due to prolonged feeding period. A sharp but reduced peak in ecdysteroid titre in the haemolymph appeared one and a half days prior to each larval ecdysis in the treated larvae, suggesting that methoprene provokes the extra larval moult through an additional release of ecdysteroids.  相似文献   

14.
Pseudomonas aeruginosa was identified as a facultative pathogen of red palm weevil. Intra-haemocoelic injection of the pathogen within larvae and pre-pupae was more effective at killing the insects [with a median lethal dose (LD50) of 9×102 to 2×103 bacteria/insect] than inoculation by force feeding (LD50 of 105 to 4×105 bacteria/insect) or by wading the insects in a suspension of the pathogen (LD50 of 105 to 2×105 bacteria/insect). Injection of 3×103 bacteria/insect killed 69% of larvae; small larvae were more susceptible (LD50 of 9×105 bacteria/larva) than either larger larvae (LD50 of 103 bacteria/larva) or pre-pupa. The median time to death of the small larvae following injection of P. aeruginosa was about 6 days but that following force feeding or wading was about 8 days. A secondary invader, Serratia marcescens, had no effect on the pathogenicity of P. aeruginosa but hastened death of larvae by about 3 days.A. Banerjee and T.K. Dangar were with the Central Plantation Crops Research Institute, Regional Station, Kayangulam 690 533, Kerala, India. They are now with the Central Rice Research Institute. Cuttack 753 006, Orissa, IndiaCPCRI research paper no. 870.  相似文献   

15.
The mode of infection and cycle of development ofTolypocladium cylindrosporum Gams was examined inAedes sierrensis andCulex tarsalis. Larvae were found to be infected through the external cuticle, the pharynx and the midgut. Blastospores and conidia were both infective although for equal numerical concentrations blastospores proved more virulent causing high mortality within the first 48 h after inoculation (80 % for L2 larvae exposed to 5×105 spores/ml), while conidia generally took 7–10 days to produce the same results. Sporulation did not occur on submerged cadavers. Conidia were produced only on floating cadavers in contact with air. Conidial production on floating 4th instar larvae was found to average 1.8×107 conidia/larva. Invasion of the haemocoele and fairly extensive growth of the fungus almost invariably occurred before larvae were killed. This was particularly true forAedes sierrensis larvae. Details are presented of growth within the host and post-mortem penetration of the fungus out of the cadaver. AdultA. sierrensis sprayed with a conidial suspension proved susceptible to infection with 100 % mortality being recorded at 10 days. Infections originated in the thorax, suggesting, the integument or possibly the thoracic spiracles to be the most probable site of infection.  相似文献   

16.
Xie  Ping  Iwakuma  Toshio  Fujii  Koichi 《Hydrobiologia》1998,368(1-3):83-90
Population dynamics of Chaoborus flavicans larvae of various instars was studied from November 1986 to December 1987 in a eutrophic, fish-free pond, Japan. First and 2nd instar larvae were observed from late April to late October, indicating a reproductive period of about half a year. C. flavicans overwintered in the 4th instar larvae. In water column samples, total density of all instars was 680–23680 m-2, and pupal density 0–2600 m-2; larvae of the 1st, 2nd, and 3rd instars showed 5–6 density peaks in 1987, suggesting that 5–6 generations occur during a year (peaks of the 4th instar larvae were not clear, probably due to their longer development than those of younger instars). In sediment samples, no 1st and 2nd instar larvae were found, 3rd instar larvae were found occasionally but density of the 4th instar larvae was 280–18600 m-2, and pupal density varied between 0–502 m-2. Fouth instar larvae accumulated in sediment in the cold season and in the water column in the warm season; high temperature and low oxygen concentration were the most important factors limiting the distribution of larvae in the sediment in summer in the NIES pond. The dry weight of total C. flavicans larvae was 0.08–4.2 g m-2 in sediment samples and 24–599 μg l-1 (0.10–2.40 g m-2) in water column samples. Comparisons of maximum densities in the NIES pond in different years and in waters of different trophic status show that density is generally higher in eutrophic than in oligotrophic habitats. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

17.
A comparison was made of the dosage-mortality responses of larvae of Pieris brassicae and P. rapae to infection by P. brassicae granulosis virus (GV). Bioassays with first, second, third, and fourth-instar larvae of both species revealed a marked difference in susceptibility between instars and between species. Median lethal dosages (LD50s) for P. rapae larvae ranged from five capsules for the first instar to 662 capsules for the fourth instar. With P. brassicae, this range extended from 66 capsules to 2.3 × 107 capsules. The time-mortality responses of the two species were similar when fed virus dosages equivalent to an LD90. Median lethal times (LT50s) ranged from 5 days for first-instar larvae to 7–8 days for fourth-instar larvae. A comparison between a long-established laboratory stock of P. brassicae and a stock recently acquired from the field showed no significant difference in their susceptibility to GV. The implications of the pronounced species differences in susceptibility to GV infection are discussed in relation to the potential field control of P. rapae and P. brassicae.  相似文献   

18.
The plant chemical azadirachtin was administered, either in artificial diet or by oral injection, to fifth instar larvae of the tobacco budworm, Heliothis virescens (Fabr.). At a dietary concentration of 0.03125 ppm, azadirachtin significantly reduced the amount of diet consumed and the weight gained by the larvae. Higher dietary concentrations (0.25 and 0.5 ppm) were necessary to reduce the efficiency of larval conversion of digested and ingested food, respectively. However, the approximate digestibility increased at the dietary concentration of 0.25 ppm.Orally injected azadirachtin (0.25 and 0.5 μg) delayed moulting to the pupal stage, produced defective pupae or adults, and inhibited development to the adult stage. Higher doses (5.0 and 10.0 μg) reduced the pre-pupal weight loss normally associated with pupation, and completely inhibited pupation. At the critical dose of 1.0 μg (the minimal dose that disrupted development to the pupal stage), azadirachtin had less of an effect on older than on younger larvae. Larvae injected on the first day of the fifth instar failed to pupate, whereas approx 40% of those injected on subsequent days pupated.The results suggest that azadirachtin affects H. virescens in a manner similar to other tested species of insects. The significance of these results, especially regarding hormonal events in the insects, is discussed.  相似文献   

19.
Larvae of the New Zealand grass grub (Costelytra zealandica) were treated with the entomopathogenic fungus, Metarhizium anisopliae, and the bacterium, Serratia entomophila, singly and in combination. The mortality of second instar larvae up to 41 days in treatments with both pathogens together was significantly greater than the additive mortalities of single pathogen treatments, and therefore synergistic. Treatment of third instar larvae with both pathogens did not increase mortality compared with the fungus alone. Second instar larvae were more resistant to M. anisopliae than third instar larvae. S. entomophila causes a chronic disease and bacterial treatments alone resulted in disease, but little mortality for either instar within 5 weeks. In both fungus alone and fungus/bacteria treatments, less than half the cadavers supported fungal sporulation. The use of a dual pathogen system for control of grass grub larvae is discussed.  相似文献   

20.
【目的】测定金龟子绿僵菌(Metarhizium anisopliae)对斜纹夜蛾(Spodoptera litura) 2龄幼虫的毒力,研究金龟子绿僵菌侵染后寄主体内抗氧化酶活性和肠道内细菌群落的变化,探讨斜纹夜蛾对金龟子绿僵菌侵染的防御机制。【方法】采用浸渍法测定不同浓度金龟子绿僵菌对斜纹夜蛾2龄幼虫的毒力;应用IlluminaMiSeq高通量测序技术测定肠道细菌群落。【结果】不同浓度的孢悬液对斜纹夜蛾2龄幼虫均有一定的毒力,处理7 d时半致死浓度(LC50)为3.944 107个孢子/mL;浓度为1.0×109个孢子/mL时,半致死时间最短(LT50)为4.6 d,校正后的死亡率为81.03%。处理后未致死的斜纹夜蛾幼虫体内抗氧化酶活性显著高于对照组。处理后致死的斜纹夜蛾幼虫肠道细菌群落多样性显著高于对照组;且处理后致死的斜纹夜蛾幼虫肠道细菌群落组成与对照组差异显著。【结论】金龟子绿僵菌对斜纹夜蛾幼虫的致死率和致死效率与金龟子绿僵菌的浓度呈正相关;斜纹夜蛾幼虫体内的抗氧化酶可能在抵抗金龟子绿僵菌侵染的过程中起重要作...  相似文献   

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