Surfactant-enhanced essential oils as mosquito larvicides

Using laboratory bioassays with fourth-instarCulex pipiens formmolestus larvae, we explored the larvicidal properties of two representative plant-derived oils, eucalyptus oil and turpentine (two grades), and cineole, the main component of eucalyptus oil. Each was larvicidal alone, but efficacy was enhanced when the spreading pressure was increased by adding 1% insoluble surfactant (Arosurf MSF) plus 1% detergent. Cheap turpentine was more effective than more refined turpentine. These mixtures were compared with the familiar surface-active larvicides Arosurf and Golden Bear Oil. At a dose of 2 μl per tub (=0.13 μl cm⁻²), enhanced by surfactants (turpentine:Arosurf:detergent 100:1:1 by volume), refined turpentine acted faster than Arosurf alone, causing higher mortality at 24 and 48 h after treatment but equivalent mortality at 72 h. It immobilised more larvae than Golden Bear Oil in the first three hours, but was less effective over 24 h. Crude turpentine enhanced by surfactants immobilised about as many larvae as Golden Bear Oil over 24 h. These findings indicate that plant essential oils merit further attention as widely available, environmentally benign mosquito larvicides.     

Enrichment of pinolenic acid from pine nut oil via lipase-catalyzed ethanolysis with an immobilized Candida antarctica lipase

Abstract

Pinolenic acid (PLA) enrichment as an ethyl ester from pine nut oil was successfully accomplished in a batch reactor by lipase-catalyzed ethanolysis using Novozym 435 lipase from Candida antarctica as a biocatalyst. PLA is predominantly an sn-3 substituent of the pine nut oil triacylglycerol (TAG), where it accounts for about 39 mol% of the fatty acids esterified at that position. In the presence of ethanol, Novozym 435 exhibited sn-3 regiospecificity with respect to the TAG of pine nut oil. The effect of the molar ratio of reactants on PLA enrichment by ethanolysis was investigated. The molar ratios of pine nut oil to ethanol were varied from 1:20 to 1:100. A fatty acid ethyl ester (FAEE) fraction with higher PLA content was obtained in the early stage of the reaction, although the yield of PLA was small. However, the PLA content of the FAEEs decreased with increasing reaction time, while the yield of PLA increased. The molar ratio of pine nut oil to ethanol that produced the optimum content and yield of PLA in FAEEs was 1:80.     

粉绿铁线莲挥发油成分分析

利用气相色谱－质谱联用技术对青海粉绿铁线莲的挥发油成分进行了研究,共鉴定出59种组分,其中主要成分为十六酸乙酯,9,12,15－十八三烯酸乙酯,亚油酸乙酯,十九烷,正二十五烷,正二十六烷,6,10,14－三甲基－2－十五烷酮,双（2－乙基己基）邻苯二甲酸酯,十八烷酸乙酯,N－苯基－萘胺等,其中十六酸乙酯的含量最高,占挥发油成分总量的24．06％,检出成分占挥发油总量的87．8％。     

Effect of frying time on free fatty acid generation and esterification rate in Aspergillus sp.-catalyzed transesterification of cottonseed oil

Abstract

The present study was carried out to examine the effect of different frying times of edible oil on the extent of transesterification catalyzed by the whole-cell biocatalyst, Aspergillus (RBD01). Cottonseed oil was chosen as a conventional and cost-effective edible oil used commercially in India. The results showed that increased frying time of the oil decreased the extent of the transesterification reaction and hence alkyl ester production. Nearly complete (>98%) transesterification to ethyl alcohol was observed with used oil containing a free fatty acid (FFA) content of 3.7%, whereas beyond an FFA content of 4.0% the yield was reduced. Biocatalyzed hydrolysis (in the absence of the ethyl alcohol acceptor) of used frying oil resulted in decreasing yield of FFA from 84.0% to 27.6% with increasing frying time. With fried oil capable of a hydrolysis yield of 82–41% FFA, transesterification reactions were nearly complete. With the lower hydrolysis yields of 38–27% FFA, the transesterified ethyl ester yield decreased to 61–51%. These observations indicate that factors other than the presence of FFA and moisture influence the biocatalytic transesterification of used cooking oils.     

Biodegradation of diesel oil and production of fatty acid esters by a newly isolated Pseudomonas citronellolis KHA

The ability of a newly isolated Pseudomonas citronellolis KHA to degrade diesel oil and to synthesize fatty acid esters has been screened in aerobic batch cultures. The microorganism was able to grow with diesel oil at initial concentrations up to 126 g/l, with optimal growth at 25 g/l. Strain KHA has produced compounds showing strong emulsifying properties (E₂₄ = 75% at the end of the exponential growth phase). The crude extract reduces the surface tension of water from 72 mN m⁻¹ down to 35 mN m⁻¹ with a corresponding minimal concentration value of 60 mg/l. GC and GC–MS analysis of crude product show that the major components are those of hexadecanoic acid propyl ester and octadecanoic acid propyl ester, which have potential for applications in cosmetics, pharmaceutical and foods industries. In addition, strain KHA represents a valuable source of compounds with surface-active properties and potential for the application in clean up of the sites contaminated with hydrocarbons.     

Inhibitory Activities of Aromatic Amino Acid Esters and Peptides against Ovalbumin Permeation through Caco-2 Cell Monolayers

Trp, Phe, and Tyr ethyl esters and their dipeptides with Gly at the C-terminals inhibited ovalbumin (OVA) permeation through Caco-2 monolayers. The inhibitory activity of Trp ethyl ester was the highest at near the concentration of 10^-6 M. It was suggested that Trp ethyl ester inhibited transcellular permeation of OVA.     

Chemical compositions and larvicidal activities of leaf essential oils from two eucalyptus species

In the current study, the mosquito larvicidal activity of leaf essential oils and their constituents from two eucalyptus species (Eucalyptus camaldulensis and Eucalyptus urophylla) against two mosquito species, Aedes aegypti and Aedes albopictus, was investigated. In addition, the chemical compositions of the leaf essential oils were analyzed using gas chromatography-mass spectrometry. Results from the larvicidal tests revealed that essential oil from the leaves of E. camaldulensis had an excellent inhibitory effect against both A. aegypti and A. albopictus larvae. The 12 pure constituents extracted from the two eucalyptus leaf essential oils were also tested individually against two mosquito larvae. Among the six effective constituents, alpha-terpinene exhibits the best larvicidal effect against both A. aegypti and A. albopictus larvae. Results of this study show that the leaf essential oil of E. camaldulensis and its effective constituents might be considered as a potent source for the production of fine natural larvicides.     

Repellency of essential oils and their components to the human body louse, Pediculus humanus humanus

Five essential oils and nine of their components were compared to diethyl toluamide (DEET) for their repellent activity against the human body louse, Pediculus humanus humanus. The absolute or intrinsic repellency of the compounds was tested by applying the repellent to corduroy patches and comparing them with untreated patches. It was found that the most effective repellents were DEET and citronella, whose activity lasted at least 29 days. The activity of rosemary lasted at least 18 days and that of eucalyptus more than 8 days. The repellent activity of the oil components such as citronellal and geraniol lasted more than 15 and 8 days, respectively. DEET remained effective at a dilution of 1:32, geraniol at 1:8, citronella at 1:4 and rosemary and citronellal at 1:1. The comparative or standard repellency of the candidate repellents was examined with the aid of a new screening technique using hairs treated with ammonium bicarbonate which is attractive to lice. Using this technique it could be shown that the repellent activity of citronella and geraniol lasted 2 days and that of rosemary and citronellal for only one day. DEET was active for less than one day. Serial dilutions of these substances also revealed that citronella was the most potent repellent for lice, followed by citronellal, rosemary, geraniol and DEET. The differences however, were not significant.     

Insecticidal activity of selected monoterpenoids and rosemary oil to Agriotes obscurus (Coleoptera: Elateridae)

Acute toxicities of three naturally occurring monoterpenoid essential oil constituents and the essential oil of rosemary were tested against late instars of Agriotes obscurus (L.) (Coleoptera: Elateridae). Both contact and volatile toxicities of thymol, citronellal, eugenol, and rosemary oil were determined. Also, phytotoxicity of these compounds was evaluated on corn germination and seedling development. Thymol had the greatest contact toxicity (LD50 = 196.0 microg/larva), whereas citronellal and eugenol were less toxic (LD50 = 404.9 and 516.5 microg/larva, respectively). Rosemary oil did not show any significant contact toxicity, even at 1,600 microg/larva. In terms of volatile toxicity, citronellal was the most toxic to wireworm larvae (LC50 = 6.3 microg/cm3) followed by rosemary oil (LC50 = 15.9 microg/cm3), thymol (LC50 = 17.1 microg/cm3), and eugenol (LC50 = 20.9 microg/cm3). Thymol, eugenol, and citronellal significantly inhibited corn seed germination and development, whereas rosemary oil had only minimal phytotoxic effects.     

Surplus Killing by Predatory Larvae of <Emphasis Type="Italic">Corethrella appendiculata</Emphasis>: Prepupal Timing and Site-Specific Attack on Mosquito Prey

Surplus or ‘wasteful’ killing of uneaten prey has been documented in the fourth larval instar of various species of the mosquito genus Toxorhynchites that occur in treeholes and other phytotelmata. Here we document surplus killing by the predatory midge Corethrella appendiculata, which in Florida cohabits treeholes and artificial containers with larvae of Toxorhynchites rutilus. Provided with a surfeit of larval mosquito prey, surplus killing was observed only in the fourth instar of C. appendiculata, peaking in intensity in the final 24 h prior to pupation, as observed for Toxorhynchites spp. Attack sites identified from videotaped encounters with mosquito prey were divided among head, thorax, abdomen, and siphon. Consumed mosquito larvae (n = 70) were attacked primarily on the head (46%) or siphon (34%), but surplus-killed prey (n = 30) were attacked predominantly on the thorax (83%). Despite its independent evolution among different insect species in aquatic container habitats, the functional significance of prepupal surplus killing remains unclear.     

Conjugates of 3-phenyllactic acid and tryptophan enhance root-promoting activity without adverse effects in Vigna angularis

3-Phenyllactic acid (PLA) is a common secondary product of Lactobacillus sp. and promotes adventitious-root formation in Azuki beans (Vigna angularis). Root promotion activity of PLA is synergistically enhanced by tryptophan (Trp). In this study, stereoisomers of PLA and Trp amide conjugates and their alkyl esters were synthesized to investigate the structure–activity relationships on root-promotion activity. The rooting activity of D-PLA-L-Trp conjugate shows more than 40 times higher than that of the mixture of D-PLA and L-Trp. Modification of PLA-Trp with ethyl ester showed the highest activity at 3,400 times of a mixture of D-PLA and L-Trp. However, L-or D-PLA-D-Trp conjugate and the isopropyl ester of PLA-Trp conjugates, both lost the root promotion activity and implicated that a requirement for steric structure for PLA related root promotion mechanism. Unlike auxin substances, which are commonly used as rooting agents that displayed high activity in low concentrations, PLA-Trp ethyl ester exhibited far less phytotoxicity at high concentration of 1 mM, despite its high rooting activity. Innovation of PLA-Trp ethyl ester may be expected for agricultural aspects with low environmental impact.     

桉树精油及其活性物质对干鱼害虫白腹皮蠹的熏蒸毒性

为了评估桉树精油和α-蒎烯对白腹皮蠹Dermestes maculatus Degeer的杀虫作用,采用生测方法测定了不同剂量和不同处理时间下,其幼虫和成虫的死亡率。结果表明:用桉树精油及其活性成分α-蒎烯处理白腹皮蠹成虫和幼虫后,可显著影响其死亡率（P＜0.001）。白腹皮蠹的反应因化学物种类、剂量和暴露时间不同而异。当剂量为32 μL/cm³的桉树精油处理白腹皮蠹幼虫72 h后,死亡率可超过90%。同样的剂量对其成虫熏蒸72 h后也表现高毒。在相同的剂量和相同的暴露时间下,α-蒎烯对白腹皮蠹的成虫和幼虫也表现高毒,且成虫的抗性比幼虫强。Probit检验结果表明桉树精油比α-蒎烯更有效。以上结果说明这些天然产物对白腹皮蠹种群的控制是很有用的。     

产谷氨酰胺菌株的诱变育种

以黄色短杆菌为出发菌株 ,经过甲基磺酸乙酯化学诱变处理 ,最后得到 1株变异株CAW 1。经初筛、复筛和传代实验 ,表明其是稳定的变异株。其谷氨酰胺的产量由诱变前的 2 .86 %提高到 4.5 8% ,提高了 6 0 .1%。     

Chromatographic separation of asparagine-linked oligosaccharides labeled with an ultravioletabsorbing compound,p-aminobenzoic acid ethyl ester

We have expanded on the suitability ofp-aminobenzoic acid ethyl ester as an ultraviolet-absorbing reagent [Wanget al., (1984) Anal Biochem 141:366–81] for the analysis of asparagine-linked oligosaccharides derived from glycoproteins. The oligosaccharides released from glycoproteins by hydrazinolysis/N-reacetylation were derivatized withp-aminobenzoic acid ethyl ester and the derivatives were purified and separated into neutral and acidic oligosaccharides on a PRE-SEP C₁₈ cartridge. The acidic oligosaccharides could be further separated into a few species by high-voltage paper electrophoresis. p-Aminobenzoic acid ethyl ester derivatives of neutral oligosaccharides were analyzed by gel permeation chromatography on Bio-Gel P-4 and HPLC on a silica-based amide column. The elution profile and the proportion of the oligosaccharides were in agreement with literature values. The overall yield of oligosaccharides from glycoproteins was approximately 70%. Fifty pmol of oligosaccharide were detectable on Bio-Gel P-4 and 4–5 pmol on HPLC.Abbreviations HPLC high performance liquid chromatography - NABEE p-aminobenzoic acid ethyl ester - FAB-MS fast-atom bombardment mass spectrometry - (GlcNAc)₂, (GlcNAc)₃, (GlcNAc)₄, (GlcNAc)₅ and (GlcNAc)₆ chito-oligosaccharides containing 2,3,4,5 and 6 residues ofN-acetylglucosamine     

Development of low-cost media for the culture of mosquito larvicides, Bacillus sphaericus and Bacillus thuringiensis serovar. israelensis

Mosquito larvicides like Bacillus sphaericus and Bacillus thuringiensis serovar. israelensis have been widely and effectively used in mosquito control programs, but the industrial production of these bacilli is expensive. Here we have attempted to develop three cost-effective media, based on cheap sources, potato, common sugar and bengalgram. Growth and production of the insecticidal proteins from these bacteria were satisfactory. Bioassay studies with different mosquito larvae showed considerable toxicity. Therefore the investigation suggests that potato-based culture media are more economical for the industrial production of B. sphaericus and B. thuringiensis serovar. israelensis.     

Insecticidal and biological efficacy of two vegetable oils against Callosobruchus maculatus (Fabricius) (Coleoptera: Bruchidae) under laboratory conditions

Bioefficacy of two vegetable oils, namely ethyl oleate and Acorus calamus, were conducted against the pulse beetle Callosobruchus maculatus (Fabr.) under a constant temperature of 29 ± 1 °C and a relative humidity of 65 ± 5% R.H. Four concentrations (0.125, 0.25, 0.5 and 1.0%) of each oil were used against the adult and tested their latent effect on the immature stages of the beetle. The results obtained show insecticidal effect of both ethyl oleate and A. calamus oil; however, A. calamus oil proved to be more toxic to the adults at all the concentrations tested in comparison to the ethyl oleate oil. As the concentration of the oils decreased, the percentage mortality decreased. The obtained results concentrated on biological efficacy of the oils tested against the adults. Both oils affected the longevities of both male and female C. maculatus adults in comparison to the control. The oviposition periods of the adult female were affected significantly in comparison to the control. A. calamus oil prohibited oviposition completely at high concentrations of 0.5 and 1.0% and caused complete sterility to the females. Lower concentrations decreased the oviposition period greatly and reduced the number of eggs laid in comparison to the control. Ethyl oleate had an inconsiderable latent effect on the incubation period as well as the percentage of hatching of the resulting eggs. A. calamus oil decreased the percentage of egg hatchability with serious increase in the percent sterility. The two tested oils had slight extension in either larval or pupal stage of the resulting progeny of C. maculatus. Both oils show oviposition deterrency towards C. maculatus adults. The oviposition deterrent index was 100% when mung bean seeds were treated with A. calamus oil. The bruchid females laid all eggs on control seeds and no eggs were laid on treated seeds.     

Urban Wet Environment as Mosquito Habitat in the Upper Midwest

Sampling of Culex larval habitat plays an important role in West Nile virus surveillance and control programs. Although many cities have established mosquito sampling programs and abatement districts, there is relatively little information describing the extent and ecology of urban surface waters and stormwater systems in different geographic areas and how these parameters affect mosquito communities and control strategies. An aerial survey of the city of Madison, Wisconsin revealed 521 above-ground wet sites. These included both constructed stormwater systems (ditches, retention ponds, detention ponds) and natural wetlands (marshes, flood areas, creeks, and rivers). Repeat sampling of 351 of these sites was conducted during 2004 and 2005. The majority of sites, 58% in 2004 and 72% in 2005, yielded no mosquito larvae, suggesting that physical and biological features of these wet sites limit the development of mosquito larvae. For both years, analysis of the positive samples revealed that less than 25% of sites produced Culex spp. while a small number of ditches and detention ponds were consistent “superproducers” of Culex larvae from year to year. This information will facilitate comparisons across geographical areas and provides insight into local variation in the public health risk due to mosquito transmission of human disease agents. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Indirect effects of the fairy shrimp,Branchipus schaefferi and two ostracod species on Bacillus thuringiensis var Israelensis-induced mortality in mosquito larvae

Bacillus thuringiensis var israelensis (BTI), when ingested by mosquito larvae, is highly toxic to them. Many other aquatic invertebrates feed on bacteria but, in general, BTI is not toxic to them. We tested in the laboratory the hypothesis that certain crustaceans indirectly benefit mosquito larvae by reducing mortality caused by BTI. We presumed the mechanism to be ingestion of the bacteria by the crustaceans resulting in a lower concentration available to the mosquito larvae.Mortality of Aedes aegypti larvae exposed to BTI was reduced in the presence of the fairy shrimp, Branchipus schaefferi and the ostracod, Cypridopsis vidua (only during summer trials and not autumn trials for the latter species) but was not reduced in the presence of the ostracod, Heterocypris incongruens. By contrast, H. incongruens preyed upon infected, though still-moving, larvae. Feeding on the bacteria by the crustaceans may not be an important mechanism; our data indicates that the crustaceans did not reduce BTI in the water. Moreover, B. schaefferi, introduced into water and then removed prior to the introduction of BTI and mosquitoes, also reduced mosquito mortality.The mechanism for the protective effect of B. schaefferi and C. vidua is unknown. However, these results suggest that the abundance of certain organisms co-occurring with mosquito larvae may partially explain why the effective concentration of BTI varies among habitats.     

Bacillus sphaericus Mtx1 and Mtx2 toxins co-expressed in Escherichia coli are synergistic against Aedes aegypti larvae

Mtx1 and Mtx2 are mosquitocidal toxins produced by some strains of Bacillus sphaericus during vegetative phase of growth. Mtx1 from B. sphaericus 2297 shows higher toxicity against Culex quinquefasciatus larvae than to Aedes aegypti larvae whereas Mtx2 from B. sphaericus 2297 shows lower toxicity against C. quinquefasciatus than to A. aegypti larvae. To test synergism of these toxins against A. aegypti larvae, mtx1 and mtx2 genes were cloned into a single plasmid and expressed in Escherichia coli. Cells producing both Mtx1 and Mtx2 toxins exhibited high synergistic activity against A. aegypti larvae approximately 10 times compared to cells expressing only a single toxin. Co-expression of both toxins offers an alternative to improve efficacy of recombinant bacterial insecticides. There is a high possibility to develop these toxins to be used as an environmentally friendly mosquito control agent.     

（S）-（-）-β-羟基苯丙酸乙酯的微生物法合成

采用酿酒酵母CGMCC No．2266菌体,不对称还原β-羰基苯丙酸乙酯制备光学纯（S）-（-）-β-羟基苯丙酸乙酯。结果表明：采用初始pH为8．0的液体发酵培养基培养的CGMCCNo．2266菌体经过50℃预热处理30min后用于生物转化获得的（S）-（-）-G-羟基苯丙酸乙酯对映体过剩值可以达到100％ee。确定了合成（S）-（-）-β-羟基苯丙酸乙酯的较佳转化条件为pH7．0,温度30℃,转化时间24h,底物浓度为3．63mmol／L,菌体用量为86g/L（干重／反应体积）。以10％葡萄糖为辅助底物,产率比不加辅助底物时提高了75．4％。在最佳转化条件下反应转化率及（S）-（-）-β-羟基苯丙酸乙酯对映体过剩值可分别达到98．4％和100％ee。