Chemical Composition,Antibacterial, Schistosomicidal,and Cytotoxic Activities of the Essential Oil of Dysphania ambrosioides (L.) Mosyakin & Clemants (Chenopodiaceae)

We have investigated the chemical composition and the antibacterial activity of the essential oil of Dysphania ambrosioides (L.) Mosyakin & Clemants (Chenopodiaceae) (DA‐EO) against a representative panel of cariogenic bacteria. We have also assessed the in vitro schistosomicidal effects of DA‐EO on Schistosoma mansoni and its cytotoxicity to GM07492‐A cells in vitro. Gas chromatography (GC) and gas chromatography‐mass spectrometry (GC/MS) revealed that the monoterpenes cis‐piperitone oxide (35.2%), p‐cymene (14.5%), isoascaridole (14.1%), and α‐terpinene (11.6%) were identified by as the major constituents of DA‐EO. DA‐EO displayed weak activity against Streptococcus sobrinus and Enterococcus faecalis (minimum inhibitory concentration (MIC) = 1000 μg/ml). On the other hand, DA‐EO at 25 and 12.5 μg/ml presented remarkable schistosomicidal action in vitro and killed 100% of adult worm pairs within 24 and 72 h, respectively. The LC₅₀ values of DA‐EO were 6.50 ± 0.38, 3.66 ± 1.06, and 3.65 ± 0.76 μg/ml at 24, 48, and 72 h, respectively. However, DA‐EO at concentrations higher than 312.5 μg/ml significantly reduced the viability of GM07492‐A cells (IC₅₀ = 207.1 ± 4.4 μg/ml). The selectivity index showed that DA‐EO was 31.8 times more toxic to the adult S. mansoni worms than GM07492‐A cells. Taken together, these results demonstrate the promising schistosomicidal potential of the essential oil of Dysphania ambrosioides.     

Influence of light spectra and elicitors on growth and ascaridole content using in vitro cultures of Dysphania ambrosioides L.

Dysphania ambrosioides L. is a medicinal plant with anti-helmintic potential. The aim of this study was to evaluate separately the effect of light spectra and elicitors on Dysphania ambrosioides growth and volatile constituents in vitro. Thus, plantlets were first cultured under blue (B), red (R), white, combinations of B:R (1:1, 2:1, 1:2) from LEDs and fluorescent lamps. Secondly, nodal segments were inoculated in the medium supplemented with chitosan (0, 50, 100, 150, and 200 mg L^??1) and salicylic acid (0, 3, 6, 9, and 12 mg L^??1). After 40 days of cultivation, the growth parameters and chemical composition of volatile constituents were evaluated. The light spectra significantly influenced in vitro growth of D. ambrosioides. The best growth occured using white LED or a blue:red combination of 2:1. It was also observed that the blue LEDs inhibited the synthesis of Z-ascaridole, while fluorescent light promoted a greater conversion of α-terpinene into ascaridole. The elicitors, chitosan and salicylic acid had a negative effect on the growth of nodal segments. However, the highest Z-ascaridole content was obtained at 50 to 100 mg L^??1 of chitosan and with 6 to 9 mg L^??1 of salicylic acid. The present study demonstrates that shoots regenerated from nodal segments exposed to different light spectra or on MS medium containing chitosan and salicylic acid can exhibit an altered growth and increased volatile constituents of interest.

     

Nematicidal activity of fervenulin isolated from a nematicidal actinomycete, <Emphasis Type="Italic">Streptomyces</Emphasis> sp. CMU-MH021, on <Emphasis Type="Italic">Meloidogyne incognita</Emphasis>

An isolate of the actinomycete, Streptomyces sp. CMU-MH021 produced secondary metabolites that inhibited egg hatch and increased juvenile mortality of the root-knot nematode Meloidogyne incognita in vitro. 16S rDNA gene sequencing showed that the isolate sequence was 99% identical to Streptomyces roseoverticillatus. The culture filtrates form different culture media were tested for nematocidal activity. The maximal activity against M. incognita was obtained by using modified basal (MB) medium. The nematicidal assay-directed fractionation of the culture broth delivered fervenulin (1) and isocoumarin (2). Fervenulin, a low molecular weight compound, shows a broad range of biological activities. However, nematicidal activity of fervenulin was not previously reported. The nematicidal activity of fervenulin (1) was assessed using the broth microdilution technique. The lowest minimum inhibitory concentrations (MICs) of the compound against egg hatch of M. incognita was 30 μg/ml and juvenile mortality of M. incognita increasing was observed at 120 μg/ml. Moreover, at the concentration of 250 μg/ml fervenulin (1) showed killing effect on second-stage nematode juveniles of M. incognita up to 100% after incubation for 96 h. Isocoumarin (2), another bioactive compound produced by Streptomyces sp. CMU-MH021, showed weak nematicidal activity with M. incognita.     

Chalcone analogues: Synthesis,activity against Meloidogyne incognita,and in silico interaction with cytochrome P450

To contribute the development of new products to control plant‐parasitic nematodes, 12 chalcone analogues were synthesized and screened for activity against Meloidogyne incognita. Three caused mortality greater than negative controls in second‐stage juvenile M. incognita, with values varying from 19.9% to 100%. The most active chalcone analogue was (1E,4E)‐1,5‐di(4‐nitrophenyl)‐2‐butylpenta‐1,4‐dien‐3‐one (compound 6 ), which had an LC₅₀ value of 41 µg/ml. Under the same conditions, the commercial nematicide Carbofuran^® (2,2‐dimethyl‐2,3‐dihydro‐1‐benzofuran‐7‐yl methylcarbamate) presented an LC₅₀ equal to 101 µg/ml. When this chalcone analogue was applied to tomato plants infested with M. incognita, reductions in the numbers of galls and eggs of 51% and 68% were observed, respectively. According to in silico studies, the enzyme target of compound 6 in M. incongita is cytochrome P450, which is important for the oxidation of several substances in the nematode. Therefore, compound 6 is potentially useful for the development of new products to control M. incongita.     

Evaluation of insecticidal activity of the essential oil of Allium chinense G. Don and its major constituents against Liposcelis bostrychophila Badonnel

Water-distilled essential oil from the dried bulbs of Allium chinense (Liliaceae) was analyzed by gas chromatography–mass spectrometry (GC–MS). Eighteen compounds, accounting for 98.4% of the total oil, were identified and the main components of the essential oil of A. chinense were methyl allyl trisulfide (30.7%), dimethyl trisulfide (24.1%), methyl propyl disulfide (12.8%) and dimethyl disulfide (9.6%) followed by methyl allyl disulfide (3.4%) and methyl propyl trisulfide (3.6%). The essential oil exhibited contact toxicity against the booklice (Liposcelis bostrychophila) with an LC₅₀ value of 441.8 μg/cm² while the two major constituents, dimethyl trisulfide and methyl propyl disulfide had LC₅₀ values of 153.0 μg/cm² and 738.0 μg/cm² against the booklice, respectively. The essential oil of A. chinense possessed strong fumigant toxicity against the booklice with an LC₅₀ value of 186.5 μg/l while methyl allyl trisulfide (LC₅₀ = 90.4 μg/l) and dimethyl trisulfide (LC₅₀ = 114.2 μg/l) exhibited stronger fumigant toxicity than methyl propyl disulfide (LC₅₀ = 243.4 μg/l) and dimethyl disulfide (LC₅₀ = 340.8 μg/l) against the booklice. The results indicated that the essential oil and its major constituents have potential for development into natural insecticides or fumigants for control of insects in stored grains.     

Mosquitocidal and repellent activities of essential oils against Culex pipiens L.

Plants are a prospective source of novel natural insect repellents and botanical insecticides. This study was conducted to investigate the chemical composition of the essential oils of three plants growing in Saudi Arabia, namely Ducrosia anethifolia, Achillea fragrantissima, and Teucrium polium; and to evaluate their potential mosquitocidal and repellent activities against adult female Culex pipiens L. The main components of the three oils were found to be decanal (28.9%) and chrysanthenyl acetate (10.04%), (D. anethifolia); sabinyl acetate (35.79) and artemesia ketone (18.28%) (A. fragrantissima); α‐cadinol (49.53%) and δ‐cadinene (10.23%) (T. polium). The oil of A. fragrantissima was the most toxic (LC₅₀ = 0.11 μL/L air) followed by D. anethifolia and T. polium with LC₅₀ values of 5.22 and 25.98 μL/L air, respectively. T. polium oil was the most repellent (292 min at 2 μL/cm²), followed by D. anethifolia and A. fragrantissima. The results indicate that the essential oils have a potential fumigant insecticidal and repellent activities for mosquito control.     

Chemical composition and larvicidal activity of essential oils from Piper species

The larvicidal activity of essential oils of four species of Piper from the Amazon Forest was tested using third-instar larvae of Aedes aegypti. The oils were extracted by steam distillation and analyzed by GC and GC–MS. The main components isolated from each Piper species were as follows: viridiflorol (27.50%), aromadendrene (15.55%) and β-selinene (10.50%) from Piper gaudichaudianum; β-selinene (15.77%) and caryophyllene oxide (16.63%) from Piper humaytanum; dillapiol (54.70%) and myristicin (25.61%) from Piper permucronatum; and asaricin (27.37%) and myristicin (20.26%) from Piper hostmanianum. Amongst all essential oils tested, the most active against larvae of A. aegypti was the oil extracted from P. permucronatum, with a LC₅₀ = 36 μg/ml (LC₉₀ = 47 μg/ml), followed by the essential oil of P. hostmanianum, with a LC₅₀ = 54 μg/ml (LC₉₀ = 72 μg/ml). The oils with higher content of arylpropanoids were more active against larvae of A. aegypti.     

Effects of culture filtrates from the nematophagous fungus Verticillium leptobactrum on viability of the root-knot nematode Meloidogyne incognita

Filtrates of three isolates of the nematophagous fungus Verticillium leptobactrum were evaluated for their nematicidal activity against the root-knot nematode Meloidogyne incognita. The filtrates inhibited egg hatching, with maximum toxicity observed for isolate HR21 at 50% (v:v) dilution, after 7 days exposure. Filtrates also inactivated second-stage juveniles (J2) at 10-50% dilutions. A scanning electron microscopy study of treated eggs showed severe alterations caused by the filtrate of isolate HR43 on M. incognita eggs, which appeared collapsed and not viable, suggesting the production of chitin-degrading enzymes or other active compounds.     

Chemical Constituents and Insecticidal Activities of Ajania fruticulosa Essential Oil

The insecticidal activity and chemical constituents of the essential oil from Ajania fruticulosa were investigated. Twelve constituents representing 91.0% of the essential oil were identified, and the main constituents were 1,8‐cineole ( 41.40% ), (+)‐camphor ( 32.10% ), and myrtenol (8.15%). The essential oil exhibited contact toxicity against Tribolium castaneum and Liposcelis bostrychophila adults with LD₅₀ values of 105.67 μg/adult and 89.85 μg/cm², respectively. The essential oil also showed fumigant toxicity against two species of insect with LC₅₀ values of 11.52 and 0.65 mg/l, respectively. 1,8‐Cineole exhibited excellent fumigant toxicity (LC₅₀ = 5.47 mg/l) against T. castaneum. (+)‐Camphor showed obvious fumigant toxicity (LC₅₀ = 0.43 mg/l) against L. bostrychophila. Myrtenol showed contact toxicity (LD₅₀ = 29.40 μg/cm²) and fumigant toxicity (LC₅₀ = 0.50 mg/l) against L. bostrychophila. 1,8‐Cineole and (+)‐camphor showed strong insecticidal activity to some important insects, and they are main constituents of A. fruticulosa essential oil. The two compounds may be related to insecticidal activity of A. fruticulosa essential oil against T. castaneum and L. bostrychophila.     

Insecticidal and feeding deterrent activities of essential oils in the cabbage looper,Trichoplusia ni (Lepidoptera: Noctuidae)

Ten essential oils were tested against the cabbage looper, Trichoplusia ni larvae for contact, residual and fumigant toxicities and feeding deterrent effects. Against third instar T. ni, Syzygium aromaticum (LD₅₀ = 47.8 μg/larva), Thymus vulgaris (LD₅₀ = 52.0 μg/larva) (the two positive controls) and Cinnamomum glanduliferum (LD₅₀ = 76.0 μg/larva) were the most toxic via topical application. Litsea pungens (LD₅₀ = 87.1 μg/larva), Ilex purpurea (LD₅₀ = 94.0 μg/larva), Cinnamomum cassia (LD₅₀ = 101.5 μg/larva) and Litsea cubeba (LD₅₀ = 112.4 μg/larva) oils were equitoxic. Thymus vulgaris (LC₅₀ = 4.8 mg/ml) and S. aromaticum (LC₅₀ = 6.0 mg/ml) oils were the most toxic in residual bioassays. Cymbopogon citratus (LC₅₀ = 7.7 mg/ml) and C. cassia (LC₅₀ = 8.5 mg/ml) oils were equitoxic followed by Cymbopogon nardus (LC₅₀ = 10.1 mg/ml) in this bioassay. The remaining five oils showed little or no residual effects. In a fumigation bioassay, L. cubeba (LC₅₀ = 16.5 μl/l) and I. purpurea (LC₅₀ = 22.2 μl/l) oils were the most toxic. Cinnamomum glanduliferum (LC₅₀ = 29.7 μl/l) and Sabina vulgaris (LC₅₀ = 31.2 μl/l) oils were equitoxic. Interestingly, S. aromaticum did not exhibit any fumigant toxicity. Cymbopogon citratus, C. nardus and C. cassia strongly deterred feeding by third instar T. ni (DC₅₀s = 26.9, 33.8 and 39.6 μg/cm², respectively) in a leaf disc choice bioassay. The different responses of T. ni larvae to the oils in different bioassays suggest that these essential oils exhibit different modes of action. Based on their comparable efficacy with essential oils already used as active ingredients in many commercial insecticides (i.e. clove oil and thyme oil), some of these essential oils may have potential as botanical insecticides against T. ni.     

Synergistic/potentiation interaction between nematostatic constituents from Azadirachta indica,Madhuca indica and Sapindus mukorossi

Triterpenic saponins isolated from seeds of Madhuca indica and fruit pericarp from Sapindus mukorossi exhibited inhibitory effect against two phyto-parasitic nematodes. Azadirachtin and salanin-nimbin-desacetylnimbin (SND) was extracted from seeds and oil of Azadirachta indica A. Juss, respectively. M. indica and S. mukorossi saponins were found to inhibit the movement of pre-adult (J₄) stage of Rotylenchulus reniformis with LC₅₀ of 168.8 and 181.9 µg/mL. Azadirachtin and SND affected the mobility of secondary juvenile stage (J₂) of Meloidogyne incognita by 83.3 and 80.1% respectively, at 0.5 mg m/L. M. indica saponin (LC₅₀ 220 µg/mL) exhibited a potentiation effect in the presence of azadirachtin in a 1:3 ratio (LC₅₀ 120.1 µg/mL). A binary mixture (1:1) of azadirachtin and SND was found to show significant nematicidal activity against M. incognita (LC₅₀ 70.9 µg/mL) and R. reniformis (LC₅₀ 91.2 µg/mL).     

Nematicidal and Molecular Docking Investigation of Essential Oils from Pogostemon cablin Ecotypes against Meloidogyne incognita

Root-knot nematode, Meloidogyne incognita is one of the most destructive nematodes worldwide. Essential oils (EOs) are being extensively utilized as eco-benign bionematicides, although the precise mechanism of action remains unclear. Pogostemon cablin Benth. is well-known as “Patchouli”. It is native to South East Asia and known for ethno-pharmacological properties. In this study, chemical composition and potential nematicidal effect of EOs hydrodistilled from the leaves of P. cablin grown at three different locations in India were comprehensively investigated to correlate their mechanism of action for target specific binding affinities toward nematode proteins. Aromatic volatile Pogostemon essential oils (PEO) from Northern India (PEO-NI), Southern India (PEO-SI) and North Eastern India (PEO-NEI) were analyzed by Gas Chromatography-Mass Spectrometry (GC/MS) to characterize forty volatile compounds. Maximum thirty-three components were identified in PEO-NEI. Sesquiterpenes were predominant with higher content of α-guaiene (2.3–24.4 %), patchoulol (6.1–32.7 %) and α-bulnesene (5.9–27.1 %). Patchoulol was the major component in PEO-SI (32.7±1.2 %) and PEO-NEI (29.2±1.1 %), while α-guaiene in PEO-NI (24.4±1.2 %). In vitro nematicidal assay revealed significant nematicidal action (LC₅₀ 44.6–87.0 μg mL⁻¹) against juveniles of M. incognita within 24 h exposure. Mortality increases with increasing time to 48 h (LC₅₀ 33.6–71.6 μg mL⁻¹) and 72 h (LC₅₀ 27.7–61.2 μg mL⁻¹). Molecular modelling and in silico studies revealed multi-modal inhibitive action of α-bulnesene (−22 to −13 kJ mol⁻¹) and α-guaiene (−22 to −12 kJ mol⁻¹) against three target proteins namely, acetyl cholinesterase (AChE), odorant response gene-1 (ODR1), odorant response gene-3 (ODR3). Most preferable binding mechanism was observed against AChE due to pi-alkyl, pi-sigma, and hydrophobic interactions. Structure nematicidal activity relationship suggested the presence of hydroxy group for nematicidal activity is nonessential, rather highly depends on synergistic composition of sesquiterpene hydrocarbons.     

Insecticidal activity of Origanum majorana L. essential oil as anti‐cholinergic agent

The present study was focused on exploring the presence of active compounds in Origanum majorana essential oil (OmEO), and its various knock‐down effects against the rice moth, Corcyra cephalonica. GC–MS analysis detected the existence of major compounds such as monoterpenes, cis‐β‐terpineol and terpinen‐4‐ol with the total proportion of 52.16%. Fumigant toxicity against adult and larvae was calculated with an LC₅₀ value of 11.31 and 49.83 μL/L air, respectively. The contact toxicity against adult, pupa, larvae and eggs was observed with LC₅₀ value 2.54, 0.95, 2.78, and 0.49 μL/L, respectively. Furthermore, the influential repellent behavior against adults has been observed. Acetylesterase (AChE) inhibition activity of OmEO was observed against adult and larvae of C. cephalonica with an IC₅₀ value of 35.89 and 118.54 μL/mL, respectively. Moreover, computational docking study revealed the binding affinity of Cis‐β‐terpineol and terpinen‐4‐ol towards the active binding sites of AChE. On the other hand, Fluorescence‐assisted cytometry and comet assay confirmed the cytotoxic and genotoxic effect of OmEO at various concentrations on C. cephalonica. Altogether, the results showed the knock‐down effect of OmEO against C. cephalonica, and it could be a potential biocontrol measure against the stored product pest.     

Insecticidal effects of a novel polyherbal formulation (HF7) against Culex pipiens L. (Diptera: Culicidae)

Plant secondary metabolites represent the most efficient and convenient method to control and overcome environmental pollution and insecticidal resistance. This study explored the mosquitocidal activity of the combined extract of seven plants, (HF7) extracted using a Soxhlet extractor against Culex pipiens under laboratory conditions. Exposure of the 3rd instars of Cx. pipiens to HF7 hexane extract resulted in LC₅₀:114.5 μg/mL and LC₉₀:117.0 μg/mL values after 24 h. The ovicidal activities of hexane extract against Cx. pipiens eggs were 21.6%, 48.3%, and 71.6% at 187.5, 93.7, and 46.88 μg/mL, respectively. HF7-treated larvae showed the formation of irregular blebbing of epithelial cells toward the lumen and sloughing into the gut lumen. HF7 extract resulted in 100% adulticidal mortality at the concentration of 3.7 mg/test tube after 30 min of exposure. The IC₅₀ of HF7 extract was 97.03 µg/ml against larvae, at which nuclear and morphological changes were observed. The spectroscopy spectrum of HF7 hexane extract disclosed the presence of 57 different secondary metabolites, among which the dominant compound was eugenol (32.3%). HF7 hexane extract could serve as a botanical insecticide for controlling Cx. pipiens and potentially other mosquito species.     

Nematicidal effect of an Amaranthus hypochondriacus L. cystatin (AhCPI) on the root-knot nematode Meloidogyne incognita (Kofoid and White) Chitwood

The root-knot nematode Meloidogyne incognita is one of the most damaging plant parasitic nematodes in the world. In this study, the effect of cystatin from Amaranthus hypochondriacus (AhCPI) as a potential control agent for M. incognita was explored. In vitro bioassays demonstrated that AhCPI affects the growth and development of eggs and the infectivity of juveniles (J2) of M. incognita, such as mortality and slower development, showing characteristic tissue damage. Mortality levels were quantified by Probit analysis, estimating LC₅₀s of 1.4 mg/mL for eggs and 0.028 mg/mL for J2. In planta bioassays showed that infected tomato seedlings treated with 0.056 mg/mL of AhCPI showed a 60% reduction in the number of galls, as compared with untreated J2-inoculated seedlings. Under greenhouse conditions, three applications of 10 mL of AhCPI (1.4 mg/mL) in the soil around the stem of M. incognita-infected tomato plants, reduced the number of galls by 93 ± 8%, as compared to the control M. incognita-infected plants. The application of AhCPI to the infected plants increased the yield (10.7%) of harvested tomato fruits, as compared to infected plants. These results show the potential of AhCPI for the control of M. incognita in tomato plants.     

Nematicidal Activity of Plant Essential Oils against Bursaphelenchus xylophilus (Nematoda: Aphelenchoididae)

The nematicidal activity and poisoning symptoms of 88 plant essential oils against Bursaphelenchus xylophilus were examined by an immersion bioassay. Results were compared with those of three trunk-injection nematicides: fenitrithion, levamisol hydrochloride, and morantel tartrate. As judged by 24 h LC₅₀ values, cinnamon bark oil (0.12 mg/ml) was the most effective nematicide, followed by coriander herb oil (0.14 mg/ml). Potent nematicidal activity was also observed with lemongrass, oregano, thyme red, and clove bud oils (LC₅₀, 0.57-0.88 mg/ml). Fenitrothion was ineffective (LC₅₀, > 10 mg/ml). In typical poisoning symptoms in B. xylophilus, these essential oils exerted rapid nematicidal action and the nematodes killed usually showed an extended shape, whereas levamisole hydrochloride and morantel tartrate usually exhibited semicircular and coiling shapes, respectively. The essential oils described merit further study as botanical nematicides for the control of pine wilt disease caused by B. xylophilus.     

Manganese sulfide nanospheres as mycocidal material and priming agent for fungi-infested rice seeds

Manganese sulfide nanospheres (MnS-NSs) prepared in situ via sonochemical process were characterized to investigate the effect of nanoscale MnS on antifungal potential against mycopathogens of rice viz. Fusarium verticillioides, Drechslera oryzae and Curvularia lunata. MnS-NSs dispersed in aqueous medium resulted in significant inhibition of growth of all the test fungi (ED₅₀ ≤ 18 μg/ml) at concentration much lower than standard commercial fungicides (captan and carbendazim with ED₅₀ ≥ 230 μg/ml) against respective fungi. The maximum per cent inhibition (98 ± 1.4) was recorded against C. lunata with a minimum ED₅₀ value of 14 μg/ml, followed by D. oryzae and F. verticillioides with ED₅₀ values of 17 and 18 μg/ml, respectively. Nanopriming of rice seeds with MnS-NSs at 35 μg/ml for 8 hr resulted in significant reduction in seed rot (76%) and seedling blight (89.13%) along with enhancement in percent germination, root length, shoot length, dry weight and vigour index of seedlings by 33.7%, 55%, 66.3%, 85.3% and 89.3%, respectively, compared with control. Hydropriming of seeds was found to control the seed health parameters less effectively than nanopriming.     

Fumigant toxicity of essential oil from Tanacetum balsamita L. (Compositae) against adults and eggs of Callosobruchus maculatus F. (Coleoptera: Bruchidae)

The essential oil of Tanacetum balsamita L. was tested against Callosobruchus maculatus F. at the rates of 5.12, 7.23, 10.19, 14.36 and 20.24?μl/l?air for adults and 5.3, 8.5, 10.8, 13.7 and 17.4?μl/l?air for eggs. All experiments were conducted at 27?±?1?°C and 65?±?5% relative humidity. Mortality was counted after 24, 48 and 72?h of exposure and all adults (dead and alive) were taken out of the vials and the vials were left at the same conditions for further 35?days to assess progeny production. For mortality of eggs, unhatched eggs were counted after 7?days. The adults treated with oil were more susceptible than eggs. At highest dose (20.24?μl/l?air) mortality of adults after 72?h, received 75.2%, but this amount for egg at 17.4?μl/l?air after 72?h was 49.06%. The lowest LC₅₀ value on 72?h was observed in adults (1.96?μl/l?air). Also, the lowest LC₉₅ value on 72?h was observed in adults (42.86?μl/l?air). The percentage of reduced progeny on 72?h was 50.92% at 20.24?μl/l?air. Our results suggest that T. balsamita can be used as an alternative to synthetic insecticides but higher dose may be needed.     

Acute toxic impacts of three heavy metals (copper,zinc, and cadmium) on <Emphasis Type="Italic">Diaphanosoma brachyurum</Emphasis> (Cladocera: Sididae)

Diaphanosoma brachyurum (Cladocera: Sididae) is a common limnetic species in summer-temperate and tropical water bodies. Few studies have investigated the sensitivity of D. brachyurum to toxic chemicals despite this species often being dominant in natural lakes and ponds. We performed acute toxicity tests of three heavy metals, copper (Cu), zinc (Zn), and cadmium (Cd), to D. brachyurum. For D. brachyurum, the lethal concentration (LC)₅₀ values of Cu (24-h LC₅₀ = 16.4 μg/L, 48-h LC₅₀ = 10.4 μg/L) and Zn (24-h LC₅₀ = 253.4 μg/L, 48-h LC₅₀ = 174.1 μg/L) were lower than those for D. magna, one of the most used test organisms for toxic chemicals. On the other hand, for D. brachyurum the 24-h LC₅₀ of Cd (166.4 μg/L) was much greater than that for D. magna, and the 48-h LC₅₀ of Cd (69.8 μg/L) was comparable. Our results indicate that D. brachyurum may be more strongly influenced by Zn and Cu than is D. magna. It is likely that the summer plankton community in which Diaphanosoma species is dominant is more sensitive to heavy metals than a community in which Daphnia species are dominant.