大链壶菌对白纹伊蚊及几种其它贵州蚊虫的杀灭作用的观察(英文)

采用大链壶菌在实验室内对白纹伊蚊及其它几种贵州蚊虫的幼虫进行杀灭试验。白纹伊蚊的感染率为25—89％,而致倦库蚊的感染率可达90—100％。实验结果为大链壶菌的宿主记录新增加一个属和五个种。     

水环境中的几种物质对大链壶菌灭蚊效率影响的初步研究

1991年1—4月,作者在实验室中观察了水中分别加入肝粉、葵花籽粉、田泥、淤泥、丝藻和人造沙后,对致倦库蚊幼虫的大链壶菌(Lagenidium giganteum)感染率的影响。发现除人造沙外,其他几种物质皆能明显地增加蚊虫的大链壶菌感染率。并且在一定范围内,蚊虫感染率与所加物质的量成正相关关系。据估计,每100ml水中加入6.4mg葵花籽粉或6mg肝粉分别可使蚊虫感染率增加2倍和3倍。作者认为大链壶菌在肝粉和葵花籽粉中弥漫地腐生生长,并释放大量额外的游动孢子,是这两种有机物增加蚊虫感染率的主要原因。 对田泥、淤泥和人造沙的研究表明,土壤肥力与蚊虫感染率的增加有关。丝藻则可能是由于净化水质造成有利于大链壶菌游动孢子活动的环境而增加蚊虫的感染率。     

The kinetics of Lagenidium giganteum growth in liquid and solid cultures

AIMS: Production of the mosquito biolarvacide Lagenidium giganteum in solid culture has been proposed as an economic alternative to production in liquid culture because of observations of improved shelf life and efficacy upon storage. Understanding the differences between these production systems and estimating growth rate in solid culture are important for commercialization. In order to address these needs a logistic model was developed to describe the growth kinetics of L. giganteum produced in solid and liquid cultures. METHODS AND RESULTS: Kinetic parameters in the logistic model were estimated by nonlinear regression of CO2 evolution rate (CER) and biomass data from solid and liquid cultivation experiments. Lagenidium giganteum biomass was measured using DNA extracted directly from samples. The logistic model was fit to experimental biomass and CER data with low standard errors for parameter estimates. The model was validated in two independent experiments by examining prediction of biomass using on-line CER measurements. CONCLUSIONS: There were significant differences between maximum biomass density, maintenance coefficients, and specific growth rates for liquid and solid cultures. The maximum biomass density (mg dw ml-1) was 11 times greater for solid cultivation compared with liquid cultivation of L. giganteum; however, the maintenance coefficient (mg CO2 h-1 (mg dw)-1) was six times greater for liquid cultivation than in solid cultivation. The specific growth rate at 30 degrees C was approximately 30% greater in liquid cultivation compared with solid cultivation. Slower depletion of substrate and lower endogenous metabolism may explain the longer shelf life of L. giganteum produced in solid culture. SIGNIFICANCE AND IMPACT OF THE STUDY: A simple logistic model was developed which allows real-time estimation of L. giganteum biomass from on-line CER measurements. Parameter estimates for liquid and solid cultivation models also elucidated observations of longer shelf life for production in solid culture.     

Confirmation of the safety of Lagenidium giganteum (Oomycetes: Lagenidiales) to mammals

Lagenidium giganteum Couch, a fungal parasite of mosquito larvae, is considered a promising agent for operational control. Quantitative reisolation of the fungus from rodent blood and tissue is easily accomplished because the cell walls are much more persistent than those of mammalian cells. The mycelium forms discrete, septate cells very early in its development, in marked contrast to filamentous fungi. The fungus is rapidly cleared from mice following intraperitoneal injection of large quantities of mycelium and oospores. More than 95% of viable cells are cleared within 24 h. Mortality following intravenous injection of mycelium in tail veins of mice is caused solely by embolism of heart and lung tissue. Recently revised U.S. Environmental Protection Agency protocols for testing of microbial pest control agents may have to be altered further to accommodate physically large microbial agents such as L. giganteum.     

灭蚊真菌——大链壶菌原生质体形成和再生的研究

采用1％纤维素酶与1％真菌脱壁酶混合液作脱壁酶,0.6mol/L山梨醇为渗透压稳定剂,从摇床培养的12—14小时菌龄的大链壶菌(Lagenidium giganteum)菌丝体获得原生质体。酶解3—5小时后,产量可达1.4—2.0×10~6/mL。并在双层培养基上初步实现了原生质体再生。     

Evaluation of various control agents against mosquito larvae in rice paddies in Taiwan.

A field test was conducted in rice paddies adjacent to Wufeng, Taichung County in Central Taiwan to evaluate the efficacy of control agents against mosquito larvae. The agents included Bacillus thuringienesis israelensis (Bti), two Lagenidium giganteum products (Lg product A and T), and temephos. The major mosquito species found in the rice paddies were Culex tritaeniorhynchus/vishnui and Anopheles sinensis. Compared to controls, a 7-day treatment with Bti or Lg products A and T caused overall reductions in the number of immatures (larvae and pupae) of Cx. tritaeniorhynchus/vishnui of 77.5%, 49.7%, and 21.9%, respectively, whereas temephos caused an increase of 66.9%. The overall reductions in An. sinensis were 85.4%, 8.6%, 44.6%, and 92.1%, respectively. There was no significant reduction in the number of mosquito larvae following 42 days of treatment with these agents. In summary, 1-week treatments with both biological control agents produced moderate overall reductions in mosquito larvae in rice paddies. The insecticide temephos, on the other hand, was very effective at suppressing the larvae of An. sinensis but significantly increased the number of Cx. tritaeniorhynchus/vishnui larvae in temephos-treated plots.     

大链壶菌灭庭院积水蚊虫的现场实验研究

为证实大链壶菌在贵州的现场灭蚊效果,首次选择部分孳生日纹伊蚊（Aedesalbopictus）和致倦库蚊（Culexquniquefasciatus）的积水轮胎和花园蓄水缸进行大链壶菌灭蚊实验。施放一次后,在连续观察的18日内该菌在两种水体中均表现了显著的治蚊作用。证明该菌可用于贵州的蚊虫综合治理。     

Oosporogenesis by Lagenidium giganteum: induction and maturation are regulated by calcium and calmodulin

Induction and maturation of the sexual stage (oospores) of the facultative mosquito parasite Lagenidium giganteum (Oomycetes: Lagenidiales) are complex developmental processes regulated by calcium-dependent events. Use of developmentally synchronized cultures of L. giganteum allowed stage-specific disruption of calcium metabolism. A calcium chelator (EGTA), an ionophore (chlortetracycline), and inhibitors of the calcium-binding protein calmodulin (dibucaine, trifluoperazine, chlorpromazine) disrupted several discrete developmental steps associated with oosporogenesis: induction of antheridia, gametangial fusion, meiosis, oospore wall formation, and subsequent spore maturation. Extracellular calcium is necessary for oosporogenesis to proceed normally and under some conditions magnesium has a synergistic effect with calcium on oospore induction. Results are discussed in relation to calcium mediation of fusion events in a number of model membrane and biological systems.     

TOXICITY AND GROWTH-REGULATING ACTIVITY OF A NEEM SEED KERNEL EXTRACT (AZAL-S) TO THE LARVAE OF CULEX QUINQUEFASCIATUS

Abstract Toxicity of a neem seed kernel extract (AZAL-S) was assessed under laboratory conditions against larvae of Culex quinquefasciatus . The LC₅₀ value for AZAL-S was 0.78× 10–6. Exposure of 4th instar larvae to 0. 3× 10–6 and 0. 5× 10–6 AZAL-S for 120 hours caused 46. 2% and 67. 5% mortality respectively, with various types of morphogenetic aberration. AZAL-S induced an obvious prologation of the larval period when 1st instar larvae were treated with 0. 4–1. 0× 10–6 AZAL-S for 7 days. The insect growth regulating effect of AZAL-S on mosquito larvae was similar to those reported for certain insect growth regulators.     

Effects of exogenous phospholipids on lipid composition and sporulation by three strains of Lagenidium giganteum.

The California (LGCA) and Butte Sink (LGBS) strains of the sterol auxotrophic fungus Lagenidium giganteum (Oomycetes: Lagenidiales) enter the sexual cycle on media supplemented with sterols. A third isolate of this mosquito pathogen, the North Carolina strain (LGNC), requires sterols plus phospholipids to produce oospores in vitro. Enrichment of the polar and neutral lipid fractions of the LGCA and LGBS strains with unsaturated fatty acids promoted oospore induction, and increased oospore viability. With the exception of the LGCA strain, there was no consistent relationship between phospholipid supplementation in growth media and mycelial phospholipid content.     

Crypticola clavulifera gen. et sp. nov. and Lagenidium giganteum: oomycetes pathogenic for dipterans infesting leaf axils in an Australian rain forest

The isolation of two entomopathogenic fungi from Forcipomyia marksae larvae collected in leaf axils of Colocasia macrorrhiza in northeastern Queensland rain forests is reported. An oomycete, in which the zoospores complete their development wholly within the sporangium and are discharged through a short papillar extension, is described as a new genus and species, Crypticola clavulifera. A strain of the well-known oomycete, Lagenidium giganteum, was isolated from F. marksae--the first record of this fungus infecting a member of the Ceratopogonidae. The pathogenesis and zoosporogenesis of the two fungi in mosquito and ceratopogonid larvae are described.     

Seasonal prevalence and host-seeking of mosquitoes in southeastern Republic of Korea

Adult mosquito surveillance was conducted at two sites at Gijang-gun, Busan southeastern area of the Korean peninsula, in 2004. The most frequently collected species at cow sheds were Anopheles sinensis sensu lato (s.l.) (83.4%), followed by Culex tritaeniorhynchus Giles (8.2%), Culex pipiens Coquillett (5.6%) and Aedes albopictus (Skuse) (2.4%). Adult populations of An. sinensis s.l. peaked during July (Trap Index, TI 1043.1), declining in August (TI 659.4) and September (TI 462.9) in 2004, while Cx. tritaeniorhynchus populations increased in August (TI 77.5) and September (TI 35.9). Adult Cx. tritaeniorhynchus was first collected in the second week of May (TI 4.0) and increased through August with two population peaks in the third week of July (TI 98.0) and the second week of August (TI 99.5). Armigeres subalbatus (Coquillett) was the most frequently collected mosquito at human-landing collections (30.4%) followed by An. sinensis s.l. (28.6%) and members of the Cx. pipiens complex (19.3%). Anopheles sinensis s.l. demonstrated two peaks of host-seeking activity at 22.00–23.00 hours and 02.00–03.00 hours in July, while in August the host-seeking peaks appeared at 22.00–23.00 hours and 03.00–04.00 hours. The black light trap collections of An. sinensis and Cx. tritaeniorhynchus demonstrated significant correlations to the ambient temperatures.     

大白口蘑菌株"厦3"的分离驯化研究

采用菌柄组织分离,从野生大白口蘑获得菌株“厦3”,经转管驯化后,在适宜温度下,菌丝满管时间从32d左右缩短到15d左右。以棉籽壳基质的菌丝长速最快,沟叶结缕草基质次之,杂木屑基质最慢;经覆土袋栽,沟叶结缕草基质的生物效率（155.8%）高于木屑基质（80.1%）,增产94.5%。脱袋栽,在草料上的生物效率达123.9％。     

Oviposition habitat selection for a predator refuge and food source in a mosquito

Abstract.  1. The influence of filamentous algae on oviposition habitat selection by the mosquito Anopheles pseudopunctipennis and the consequences of oviposition decisions on the diet, development, body size, and survival of offspring were examined.
2. A natural population of An. pseudopunctipennis in Chiapas, Mexico, oviposited almost exclusively in containers with filamentous algae. Algae represented 47% of the gut contents of mosquito larvae sampled from the natural population. Mosquito larvae fed on an exclusive diet of algae developed as quickly and achieved the same adult body size (wing length) as their conspecifics fed on a standard laboratory diet.
3. Multiple regression of survival of mosquito larvae on percentage surface area cover of algae (0–99%) and the density of predatory fish (zero to four fish per container) was best described by a second-order polynomial model. Increasing fish densities resulted in a reduction in mosquito survival in all algal treatments. The highest incidence of survival was observed at intermediate (66%) algal cover in all treatments.
4. The presence of fish significantly extended larval development times whereas algal cover had no significant effect. The presence of fish resulted in emergence of smaller adults due to reduced feeding opportunities and predator avoidance behaviour. Algal cover also affected mosquito wing length but differently at each fish density.
5. Oviposition habitat selection improves survival in the presence of predators and feeding opportunities for An. pseudopunctipennis larvae.     

UP REGULATION OF UBIQUITIN C-TERMINAL HYDROLASE IN THE RESPONSE OF THE MOSQUITO ANOPHELES STEPHENS TO PLASMODIUM YOELII INFECTION

Abstract The isolation and study of genes that are differentially expressed in malaria-infected mosquitoes is important for the elucidation of basic molecular mechanisms underlying vector-parasite interactions. When screening against a previously established cDNAs pool representing specifically expressed genes in the mosquito Anopheles stephemi infected by Plasnwdium γoelii , it was found that one of these encodes a protein with extensive sequence similarity to the Drosophila melarwgaster ubiquitin C-terminal hydrolase (UCTH). Similarity alignment showed that the fragment is 89% identical at amino acid level to the corresponding region of the known An. gambiae EST sequence, as well as 63 % identical to that of both the fruittly and human sequence. Virtual Northern blot expression dynamics of the gene indicated that it was up-regulated significantly in the mosquito at least 1–7 days post-infection, consistent with the critical transition stages of midgut invasion and relocation of sporozoites from the oocysts to the salivary glands during parasite development. Rather little is known about the role of the ubiquitin pathway in the activation of the mosquito innate immune system. The results indicate that the gene is related to malaria infection in mosquito. The cloning and expression profile analysis of AsUCTH enables us to make predictions as to the roles it may play during malaria infection.     

Isolation and characterization of a novel angiotensin I-converting enzyme inhibitory peptide derived from the edible mushroom Tricholoma giganteum

The fruiting body of Tricholoma giganteum has many pharmaceutical uses and has long been utilized as a home remedy in Asia. This study describes the extraction and characterization of the first angiotensin I-converting enzyme (ACE) inhibitory peptide from T. giganteum. The maximum ACE inhibitory activity (IC50: 0.31 mg) was obtained when the fruiting body of T. giganteum was extracted with distilled water at 30 degrees C for 3 h. After the purification of ACE inhibitory peptides with ultrafiltration, Sephadex G-25 column chromatography, and reverse-phase HPLC, an active fraction with an IC50 of 0.04 mg and a yield of 0.3% was obtained. The ACE inhibitory peptide was a novel tripeptide, showing very low similarity to other ACE inhibitory peptide sequences, and was sequenced as Gly-Glu-Pro. The purified ACE inhibitor from T. giganteum competitively inhibited ACE, and it maintained inhibitory activity even after incubation with proteases. ACE inhibitor from T. giganteum showed a clear antihypertensive effect in spontaneously hypertensive rats (SHR), at a dosage of 1 mg/kg.     

Studies on physiology,zoospore morphology and entomopathogenic potential of the aquatic oomycete: Lagenidium giganteum

The oomycete Lagenidium giganteum, a facultative parasite of mosquito larvae requires exogenous sterols for the genesis of zoospores when grown saprobically. Growth media prepared from oil rich materials such as soy or sunflower seed were very effective inducers of virulent zoospores. The external morphology of zoospores of L. giganteum was studied with the aid of philips scanning electron microscope 515. Zoospores were ovoid, bluntly pointed with the groove parallel to the long axis and 0.7 × 1.4 μm. Insect cell walls are known to contain lipid and chitin. L. giganteum was tested for chitinase activity and found to possess 0.76 ± SD0.14 chitinase activity. Use of oil seed for growth of the organism confirms phospholipase activity. Phospholipase production was studied further by egg-yolk plate method. Presence of these two key enzymes that can initiate host cell damage suggests the entomopathogenic potential of L. giganteum. L. giganteum failed to grow at 37 °C limiting its effectiveness in warmer climates. Introduction of this organism to variety of habitats with various mosquito species will demonstrate the efficacy of the organism as a bioinsecticide. This revised version was published online in June 2006 with corrections to the Cover Date.     

Prevalence of Aedes camptorhynchus (Thomson) (Diptera: Culicidae) and Other Mosquitoes in the Eastern Coast of Victoria

Adult mosquito populations were monitored using dry-ice baited EVS light traps at weekly intervals for four seasons (1991–95) in Avon, Bairnsdale, Rosedale and Hastings, and for three seasons (1991–94) in Tambo, in the eastern coast of Victoria. Among the 20 species of adult mosquitoes recorded, Aedes camptorhynchus (Thomson) was dominant (74–99%). Higher Ae camptorhynchus adult populations were recorded in the 1992–93 and 1993–94 seasons, than in 1991–92 and 1994–95 seasons. Rosedale shire recorded higher Ae camptorhynchus adult populations than the Avon, Bairnsdale, Hastings and Tambo shires. In all shires, two peaks of Ae camptorhynchus activity reaching high (100–1,000 adults/trap) to very high (> 1,000 adults/trap) levels were recorded, with the first peak in the middle of December (summer) and the second peak in the middle of April (autumn). the Ae camptorhynchus activity in the coastal region increased with an increase in the minimum temperature.