Yield,chemical composition,and bioactivity of essential oils from 12 species of Eucalyptus on Aedes aegypti larvae

The insecticidal activity of essential oils from 12 species of Eucalyptus (Myrtaceae) was evaluated on larvae of Aedes aegypti (L.) (Diptera: Culicidae), the most important vector of dengue and yellow fever in the Americas. Oils were obtained by hydrodistillation and their chemical composition was determined by gas chromatography coupled to mass spectrometry; yields ranged from 0.2 to 2.5%. Essential oils were mainly composed of 1,8‐cineole, α‐pinene, α‐phellandrene, β‐phellandrene, γ‐terpinene, 4‐terpineol, α‐terpineol, p‐cymene, and spathulenol. Larvicidal effects were tested on susceptible third or fourth stage Ae. aegypti larvae, determining median lethal concentration (LC₅₀) and median effective concentration (EC₅₀). Essential oils from Eucalyptus dunnii (Maiden), Eucalyptus gunnii (Hook), Eucalyptus tereticornis (Smith), Eucalyptus camaldulensis (Dehn), and Eucalyptus saligna (Smith) showed the best larvicidal activities with LC₅₀ values of 25.2, 21.1, 22.1, 26.8, and 22.2, respectively. No significant differences were observed between LC₅₀ and EC₅₀ values of the same oil. Regression analysis revealed a significant relationship between total essential oil yields and 1,8‐cineole concentration. Significant relationships were also revealed between larval mortality and the concentration of 1,8‐cineole and p‐cymene. This indicated that Eucalyptus species with high oil yields have higher 1,8‐cineole concentrations and lower p‐cymene concentrations and have less effect on Ae. aegypti. Our results suggest the potential of controlled crossing methods to obtain Eucalyptus trees with chemical profiles having enhanced activity against this mosquito.     

Volatiles from Marina neglecta: Biocide effect on insect vectors of tropical diseases in Southern Mexico

Preventive measures based in the control of insect vectors are considered as the best choice to decrease the incidence of insect-borne diseases. Herein we report on the volatile content of the leaf essential oils from Marina neglecta, a medicinal plant distributed in the tropical regions of southern Mexico. In order to investigate the chemical variation of the essential oils, a volatile screening was performed during the four seasons of the years 2016–2019. Simultaneously, their biological activity was tested on distinct life stages of Meccus pallidipennis, M. bassolsae, Aedes aegypti and A. albopictus. Essential oils were mainly constituted of β-pinene (>30%) β-caryophyllene (>25%) and germacrene D (>13%). Dorsal-abdomen application of essential oils on triatomines, revealed an efficient LC₅₀ for nymphs of the stages I to III (4 µg/insect), nymphs of the stages IV to V (5–6 µg/insect), and adults (7–8 µg/insect). The LT₅₀ for the stages I to III was between 6 and 8 h, whereas that for the stages IV to V and adults oscillated between 12 and 16 h and 22 to 26 h, respectively. Fumigation experiments performed on nymph V, demonstrated that 300 µg L^?1 air produced 100% mortality after 72 h post-treatment. Among tested volatiles, β-pinene and β-caryophyllene produced a comparable mortality rate (p < 0.01) than that of essential oils in the stages assayed. Essential oils showed strong larvicidal (LC₅₀, 24–36 µg mL^?1) and adulticidal (35–48 µg mL^?1) activities in mosquito species with an LT₅₀ of 4.5 h and 25–35 min, respectively. The evaluation of β-pinene produced a significant mortality rate (p < 0.01) in larvae whereas germacrene D was the most effective volatile (p < 0.01) against adults of both mosquito species. According to our results, β-pinene was the most effective volatile against the four insect species evaluated and its effect was comparable to that of the essential oil.     

Chemical composition and larvicidal activity of essential oils of three Artemisia species

Aedes aegypti L. (Diptera: Culicidae) is a vector for serious diseases in tropical regions. This pest is mainly controlled by commercial larvicides but the application of such products has led to environmental problems. Essential oils (EO) have been consistently reported as molecules with insecticidal activity and can be used to produce more environmentally friendly larvicides in the control of A. aegypti. In this study, the larvicidal effect of essential oils (EO) from the leaves of three Artemisia species was evaluated against A. aegypti. The oils were obtained from steam distillation and their chemical composition was determined by gas chromatography–mass spectrometry. The EO of Artemisia camphorata was the most active in the screening bioassay and presented LC₅₀ and LC₉₅ of 64.95 and 74.18 μg ml⁻¹, respectively. In addition, we found that germacrene D-4-ol was the constituent responsible for the toxicity of this EO. Artemisia camphorata EO and its major constituent, germacrene D-4-ol, are promising for the development of natural larvicides against A. aegypti.     

In vitro assays on the susceptibility of four species of nematophagous fungi to anthelmintics and chemical fungicides/antifungal drug

Using nematophagous fungi for the biological control of animal parasitic nematodes will become one of the most promising strategies in the search for alternative chemical drugs. The purpose of this study was to check the in vitro activity of four anthelmintics, four chemical fungicides and two antifungal drugs on the spore germination of nematophagous fungi: Duddingtonia flagrans (SF170), Arthrobotrys oligospora (447), Arthrobotrys superba (435) and Arthrobotrys sp. (PS011). A modified 24-well cell culture plate assay was conducted to evaluate the susceptibility of nematophagous fungi against drugs tested by calculating the effective middle concentrations (EC₅₀) of each tested drug to inhibit the germination of fungal spores. EC₅₀ ranged between 0·7 and 47·2 μg ml⁻¹ for fenbendazole, thiabendazole and ivermectin, except levamisole (546·5–4057·8 μg ml⁻¹). EC₅₀ of tested fungicides was 0·6–2·3 μg ml⁻¹ for carbendazim, 55·9–247·4 μg ml⁻¹ for metalaxyl, 24·4–45·2 μg ml⁻¹ for difenoconazole, and 555·9–1438·3 μg ml⁻¹ for pentachloronitrobenzene (PCNB). EC₅₀ of two antifungal drugs was 0·03–3·4 μg ml⁻¹ for amphotericin B and 0·3–10·9 μg ml⁻¹ for ketoconazole. The results showed that 10 tested drugs, except for levamisole and PCNB, had in vitro inhibitory effects on nematophagous fungi. The chlamydospores of D. flagrans had the highest sensitivity to nine tested drugs, except for ketoconazole.     

Mosquito larvicidal efficacy of phenolic acids of seaweed Chaetomorpha antennina (Bory) Kuetz. against Aedes aegypti

Mosquito larvicidal and repellent activities of phenolic acids of Chaetomorpha antennina (Bory) Kuetz. against the third instar larvae of Aedes aegypti were investigated. The larval mortality was observed after 24 h exposure. Results of mosquito larvicidal tests revealed that insoluble bound phenolic acids and soluble conjugated phenolic acid fractions of C. antennina had an excellent inhibitory effect against A. aegypti and its LC₅₀ values were 23.4 and 44.6 μg ml⁻¹, respectively. The repellency assay of insoluble bound phenolic acids and soluble conjugated phenolic acid fractions of C. antennina, at 10 μg cm⁻² concentration gave 100% protection up to 120 min. The results indicate that phenolic acids of C. antennina have a wide spectrum of larvicidal and repellent activities against Aedes aegypti.     

The effects of plant essential oils on escape response and mortality rate of Aedes aegypti and Anopheles minimus

The High Throughput Screening System (HITSS) has been applied in insecticide behavioral response studies with various mosquito species. In general, chemical or natural compounds can produce a range of insect responses: contact irritancy, spatial repellency, knock‐down, and toxicity. This study characterized these actions in essential oils derived from citronella, hairy basil, catnip, and vetiver in comparison to DEET and picaridin against Aedes aegypti and Anopheles minimus mosquito populations. Results indicated the two mosquito species exhibited significantly different (P<0.05) contact irritant escape responses between treatment and control for all tested compound concentrations, except with the minimum dose of picaridin (P>0.05) against Ae. aegypti. Spatial repellency responses were elicited in both mosquito species when exposed to all compounds, but the strength of the repellent response was dependent on compound and concentration. Data show that higher test concentrations had greatest toxic effects on both mosquito populations, but vetiver had no toxic effect on Ae. aegypti and picaridin did not elicit toxicity in either Ae. aegypti or An. minimus at any test concentration. Ultimately, this study demonstrates the ability of the HITSS assay to guide selection of effective plant essential oils for repelling, irritating, and killing mosquitoes.     

Synthesis and larvicidal activity of indole derivatives against Aedes aegypti (Diptera: Culicidae)

In light of the challenges to control Aedes aegypti and the critical role that it plays as arbovirus vector, it is imperative to adopt strategies that provide fast, efficient and environmentally safe control of the insect population. In the present study, we synthesized six indole derivatives (C1‐C6) and examined their larvicidal activity and persistence against Ae. aegypti larvae, as well as their toxicity towards Raw 264.7 macrophages, Vero cells, Chlorella vulgaris BR017, Scenedesmus obliquus BR003, Caenorhabditis elegans N2 and Galleria mellonella. Among the bioactive compounds (C1, C2, C4 and C5), C2 exerted the strongest larvicidal activity against Ae. aegypti, with LC50 = 1.5 μg/ml (5.88 µM) and LC90 = 2.4 μg/ml (9.50 µM), indicating that the presence of chlorine or bromine groups in the aromatic ring improved the larvicidal activity of the indole derivatives. C1, C2, C4 and C5 did not reduce viability of RAW 264.7 macrophages, Vero cells, C. elegans N2 and G. mellonella. Compounds C1, C2 and C5 did not affect the growth of C. vulgaris BR017 and S. obliquus BR003. Analysis of larvicidal persistence under laboratory conditions revealed that the effect of compounds C1, C2, C4 and C5 lasted for 30 days and caused 100% of larvae mortality within few hours. Altogether, our findings demonstrate that the indole derivatives C1, C2, C4 and C5 effectively control Ae. aegypti larvae population, without clear signs of toxicity to mammalian cells, algae, C. elegans and G. mellonella.     

Environmentally-Friendly Pesticidal Activities of Callicarpa and Karomia Essential Oils from Vietnam and Their Microemulsions

There is an ongoing interest to identify alternative pesticidal agents to avoid the chronic problems associated with synthetic pesticides. Essential oils have shown promise as botanical pest control agents. In the present study, the essential oils of four members of the Lamiaceae (Callicarpa candicans, C. erioclona, C. macrophylla, and Karomia fragrans; Vietnamese names: Nàng nàng, Tu châu lông mem, Tu châu lá to and Cà diện, respectively), obtained from wild populations in Vietnam, have been obtained by hydrodistillation and analyzed by gas chromatography-mass spectrometry. The essential oils were formulated into microemulsions and the essential oils and their microemulsions were screened for mosquito larvicidal activity against Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and for molluscicidal activity against Pomacea canaliculata. Atractylone and (E)-caryophyllene dominated the volatiles of C. candicans (CCEO) and C. erioclona (CEEO), while the major component in C. macrophylla (CMEO) and K. fragrans (KFEO) was (E)-caryophyllene. The essential oils and microemulsions of both C. candicans and C. erioclona exhibited excellent larvicidal activity against all three mosquito species (Ae. aegypti, Ae. albopictus, and Cx. quinquefasciatus) with LC₅₀ values <10 μg/mL. Additionally, the larvicidal activity of the microemulsions were significantly improved compared with their free essential oils, especially for C. candicans and C. erioclona. All four essential oils and their microemulsions showed excellent molluscicidal activity with LC₅₀ <10 μg/mL. In most cases, the essential oils and microemulsions showed greater pesticidal activity against target organisms than the non-target freshwater fish, Oreochromis niloticus. The in silico studies on physicochemical and ADMET properties of the major components in the studied essential oils were also investigated and most of the compounds possessed a favorable ADMET profile. Computational modeling studies of the studied compounds demonstrated a favorable binding interaction with the mosquito odorant-binding protein target and support atractylone, β-selinene, and caryophyllene oxide as potential inhibitors. Based on the observed pesticidal activities of the essential oils and their microemulsions, the Callicarpa species and K. fragrans should be considered for potential cultivation and further exploration as botanical pesticidal agents.     

Insights into collateral susceptibility and collateral resistance in Acinetobacter baumannii during antimicrobial adaptation

The susceptibility of Acinetobacter baumannii exposed to primary antibiotic can be either increased or decreased when exposed to secondary antibiotic. This study was designed to assess the relative fitness, collateral susceptibility and collateral resistance of polymyxin B- (PMB-) adapted A. baumannii to ciprofloxacin (CIP), meropenem (MER), PMB, tetracycline (TET) and tobramycin (TOB). Strains of wild-type A. baumannii KACC 12454 (AB^KACC), wild-type A. baumannii CCARM 12088 (AB^CCARM), PMB-adapted AB^KACC, PMB-adapted AB^CCARM, stabilized AB^KACC and stabilized AB^CCARM were used in this study. Compared to the wild-type AB^KACC, the MICs of PMB were increased from 2 to 128 μg ml⁻¹ against PMB-adapted AB^KACC, while MICs of CIP, MER, TET and TOB were decreased from 2 to 1 μg ml⁻¹, 16 to 1 μg ml⁻¹, 16 to 2 μg ml⁻¹ and 64 to 16 μg ml⁻¹, respectively. The PMB-adapted AB^CCARM was resistant to CIP (32 μg ml⁻¹) and PMB (64 μg ml⁻¹) compared to the wild-type AB^CCARM. The resistance of stabilized AB^KACC and AB^CCARM to all antibiotics was lost after antibiotic-free culture in the exception of CIP and TET. The susceptibilities of wild-type, PMB-adapted and stabilized AB^KACC and AB^CCARM to CIP, MER, PMB, TET and TOB were increased in the presence of β-lactamase and efflux pump inhibitors. The high levels of relative fitness were observed for stabilized AB^KACC, PMB-adapted AB^CCARM and stabilized AB^CCARM. The stabilized AB^KACC and PMB-adapted AB^CCARM were highly heteroresistance to PMB and TET, respectively. The PMB-adapted AB^KACC and AB^CCARM showed various antibiotic patterns, known as collateral susceptibility and collateral resistance. The results provide useful information for designing effective antibiotic regimens that can enhance the antibiotic activity against A. baumannii infections.     

Larvicidal effects of some essential oils against Aedes aegypti (L.), the vector of dengue fever in Saudi Arabia

Essential oils are very popular among organic growers because they are ecologically safe, do not have mammalian toxicity, and cannot be resistant to a variety of contaminants. Four essential oils, Lemon, Lavender, Peppermint, and Neem, were tested for larvicide efficacy against the dengue fever vector Aedes aegypti larvae under laboratory conditions using dipping bioassay techniques. Among the essential oils tested, lemon, peppermint, and lavender oils showed high larvicidal activity against larvae of Ae. aegypti. Lemon oil showed the highest effects (LC₅₀ 10.676 ppm), while Peppermint, Lavender and Neem oil showed the lowest effects (LC₅₀ 21.380, 29.818 and 38.058 ppm, respectively). As a result, the mixture of lemon oil (LC₅₀) with Peppermint oil (LC₂₅) showed the highest co-toxicity factor, whereas the mixture of Lemon oil (LC₅₀) with Diesel oil (LC₂₅) showed the lowest co-toxicity factor. Based on the results of this study, it appears that essential oils may be useful as larvicides against Ae. aegypti larvae. In search of new natural larvicides, these compounds may provide an alternative to Synthetic insecticides as these are environmentally safe insecticides.     

Isolation of antifouling compounds from the marine bacterium, <Emphasis Type="Italic">Shewanella oneidensis</Emphasis> SCH0402

Two compounds, 2-hydroxymyristic acid (HMA) and cis-9-oleic acid (COA), were isolated from a chloroform extract of the marine bacterium, Shewanella oneidensis SCH0402. In a spectrophotometer-based chemotaxis assay, HMA completely eliminated the optical density (OD) of Alteromonas marina SCH0401 and Bacillus atrophaeus SCH0408, motile, fouling bacteria, at 100 and 1000 μg ml⁻¹, respectively. COA similarly decreased the OD of A. marina and B. atrophaeus by 100% at 1000 μg ml⁻¹. The commercially available, highly toxic anti-fouling compound, tributyltin oxide (TBTO) never reduced the OD of the target bacteria by 100% even at higher concentration. Instead, all the test bacterial cells were killed at higher than 1000 μg ml⁻¹ of concentration. Both HMA and COA inhibited germination of Ulva pertusa spores completely at 10 and 100 μg ml⁻¹, respectively, while TBTO inhibited germination at 0.01 μg ml⁻¹. However, in field assays, both HMA and COA showed anti-fouling activities as potent as TBTO against a wide range of fouling organisms, including micro- and macro-algae, barnacles, and mussels. The average fouling coverage on the surface of the control panel was 93 ± 6% after 1.5 years but no fouling was observed on the surface of the test panel onto which each compound was applied separately. Thus, bacterial repellent compounds can be used as substitutes for potent toxic anti-fouling compounds, resulting in higher standards of environmental safety without loss of anti-fouling performance.     

Lantana camara Essential Oils from Vietnam: Chemical Composition,Molluscicidal, and Mosquito Larvicidal Activity

Lantana camara is a troublesome invasive plant introduced to many tropical regions, including Southeast Asia. However, the plant does hold promise as a source of essential oils that may be explored for potential use. Fresh water snails such as Pomacea canaliculata, Gyraulus convexiusculus, and Tarebia granifera can be problematic agricultural pests as well as hosts for parasitic worms. Aedes and Culex mosquitoes are notorious vectors of numerous viral pathogens. Control of these vectors is of utmost importance. In this work, the essential oil compositions, molluscicidal, and mosquito larvicidal activities of four collections of L. camara from north-central Vietnam have been investigated. The sesquiterpene-rich L. camara essential oils showed wide variation in their compositions, not only compared to essential oils from other geographical locations (at least six possible chemotypes), but also between the four samples from Vietnam. L. camara essential oils showed molluscicidal activities comparable to the positive control, tea saponin, as well as other botanical agents. The median lethal concentrations (LC₅₀) against the snails were 23.6–40.2 μg/mL (P. canaliculata), 7.9–29.6 μg/mL (G. convexiusculus), and 15.0–29.6 μg/mL (T. granifera). The essential oils showed good mosquito larvicidal activities with 24-h LC₅₀ values of 15.1–29.0 μg/mL, 26.4–53.8 μg/mL, and 20.8–59.3 μg/mL against Ae. aegypti, Ae. albopictus, and Cx. quinquefasciatus, respectively. The essential oils were more toxic to snails and mosquito larvae than they were to the non-target water bug, Diplonychus rusticus (24-h LC₅₀=103.7–162.5 μg/mL). Sesquiterpene components of the essential oils may be acting as acetylcholinesterase (AChE) inhibitors. These results suggest that the invasive plant, L. camara, may be a renewable botanical pesticidal agent.     

Evaluation of larvicidal enhanced activity of sandalwood oil via nano-emulsion against Culex pipiens and Ades aegypti

Mosquito control with essential oils is a trending strategy using aqueous oil nano-emulsions to expand their performance. Sandalwood essential oil and its prepared nano-emulsion used to estimate their larvicidal activities against the 3rd instar larvae of Culex pipiens and Aedes aegypti and their effects on larval tissue detoxifying enzymes. Sandalwood nano-emulsion was characterized by homogeneous, stable, average particles size (195.7 nm), polydispersity index (0.342), and zeta potential (?20.1 mV). Morphologically showed a regular spherical shape in size ranged from 112 to 169 nm that confirmed via scanning electron microscopy. Oil analysis identified sesquiterpene alcohols, mainly santalols, terpenoids, aromatic compounds, fatty acid methyl esters, and phenolic compounds. Larvicidal activities of the oil and its nano-emulsion indicated dose, formulation, and exposure time-related mortality after 24 and 48 h in both species. After 24 h, 100% mortality was detected at 1000 ppm for the nano-emulsion with LC₅₀ of 187.23 and 232.18 ppm and at 1500 ppm for the essential oil with an LC₅₀ of 299.47 and 349.59 ppm against the 3rd larvae Cx. pipiens and Ae. aegypti, respectively. Meanwhile, an enhanced significant effect of the nano-emulsion was observed compared to oil exposure in decreasing total protein content and the activities of alkaline phosphatase and β-esterase enzymes, and increasing α-esterase and glutathione S-transferase activities in larval body tissues. Results demonstrated the enhanced larvicidal potential of sandalwood oil nano-emulsion over that of oil. The effect involved alterations in the detoxifying enzymes based on the existing natural active ingredients against Cx. pipiens and Ae. aegypti larvae.     

Electrophysiological and behavioural response of Aedes albopictus to n‐heinecosane,an ovipositional pheromone of Aedes aegypti

Aedes aegypti (L.) and Aedes albopictus (Skuse) (Diptera: Culicidae) are highly anthropophilic mosquito species and potential vectors of dengue and yellow fever. The location of suitable sites for oviposition requires a set of visual, tactile, and olfactory cues that influence females before they lay their eggs. In this study, the effect of n‐heneicosane, a recognized oviposition pheromone of Ae. aegypti, on the olfactory receptors of the antennae of Ae. aegypti and Ae. albopictus was studied using electroantennographic detection coupled to gas chromatography (GC‐EAD). A significant electroantennographic response to n‐heneicosane in adult females of both mosquito species was observed. In addition, gravid Ae. albopictus females laid more eggs in substrate treated with n‐heneicosane at 0.1, 1, or 10 p.p.m. than in the control, denoting oviposition attractancy. Conversely, at 30, 50, 100, and 200 p.p.m., more eggs were laid in the control substrate, indicating oviposition repellency. Analysis of the larval cuticle by GC and mass spectrometry confirmed the presence of n‐heneicosane in the cuticles of Ae. albopictus larvae. The species‐specific role of n‐heneicosane as an oviposition pheromone in Ae. aegypti and its significance as a behaviour modifier of Ae. albopictus in breeding sites is discussed.     

Antibiofilm and antifungal activities of medium-chain fatty acids against Candida albicans via mimicking of the quorum-sensing molecule farnesol

Candida biofilms are tolerant to conventional antifungal therapeutics and the host immune system. The transition of yeast cells to hyphae is considered a key step in C. albicans biofilm development, and this transition is inhibited by the quorum-sensing molecule farnesol. We hypothesized that fatty acids mimicking farnesol might influence hyphal and biofilm formation by C. albicans. Among 31 saturated and unsaturated fatty acids, six medium-chain saturated fatty acids, that is, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid and lauric acid, effectively inhibited C. albicans biofilm formation by more than 75% at 2 µg ml⁻¹ with MICs in the range 100–200 µg ml⁻¹. These six fatty acids at 2 µg ml⁻¹ and farnesol at 100 µg ml⁻¹ inhibited hyphal growth and cell aggregation. The addition of fatty acids to C. albicans cultures decreased the productions of farnesol and sterols. Furthermore, down-regulation of several hyphal and biofilm-related genes caused by heptanoic or nonanoic acid closely resembled the changes caused by farnesol. In addition, nonanoic acid, the most effective compound diminished C. albicans virulence in a Caenorhabditis elegans model. Our results suggest that medium-chain fatty acids inhibit more effectively hyphal growth and biofilm formation than farnesol.     

Pathogenicity of entomopathogenic fungus,Metarhizium anisopliae MET-GRA4 isolate on dengue vectors,Aedes albopictus and Aedes aegypti mosquito larvae (Diptera: Culicidae)

Considering the rapid transmission of the dengue virus, substantial efforts need to be conducted to ward-off the epidemics of dengue viruses. The control effort is depending on chemical insecticides and had aroused undesirable conflicts of insecticide resistance. Here, we study the entomopathogenic fungus, Metarhizium anisopliae as a promising new biological control agent for vector control. The pathogenicity effects of Metarhizium anisopliae against field and laboratory strains of Aedes albopictus and Aedes aegypti larvae were tested using the larvicidal bioassay technique. The results demonstrate that the treatments using M. anisopliae isolate MET-GRA4 were highly effective and able to kill 100% of both Ae. albopictus and Ae. aegypti mosquito larvae at a conidia concentration of 1 × 10?/ml within 7 days of the treatment period. The fungus displayed high larvicidal activity against laboratory and field strain of Ae. aegypti larvae with LC₅₀ values (9.6 × 10³/ml, 1.3 × 10³/ml) and LC₉₅ values (1.2 × 10?/ml, 5.5 × 10⁵/ml) respectively. For Ae. albopictus, LC₅₀ values for laboratory and field strains were (1.7 × 10⁴/ml, 2.7 × 10⁴/ml) and the LC₉₅ values were (2.1 × 10?/ml, 7.0 × 10⁵/ml) respectively. Interestingly, the susceptibility of field strain towards M. anisopliae was higher as compared to the laboratory strain Aedes larvae. In which, the causative agents of all the dead larvae were verified by the virulence of M. anisopliae and caused morphological deformities on larval body. The findings from this study identify this isolate could be an effective potential biocontrol agent for vector mosquitoes in Malaysia.     

Chemical composition and larvicidal activity of edible plant‐derived essential oils against the pyrethroid‐susceptible and ‐resistant strains of Aedes aegypti (Diptera: Culicidae)

The chemical compositions and larvicidal potential against mosquito vectors of selected essential oils obtained from five edible plants were investigated in this study. Using a GC/MS, 24, 17, 20, 21, and 12 compounds were determined from essential oils of Citrus hystrix, Citrus reticulata, Zingiber zerumbet, Kaempferia galanga, and Syzygium aromaticum, respectively. The principal constituents found in peel oil of C. hystrix were β‐pinene (22.54%) and d‐limonene (22.03%), followed by terpinene‐4‐ol (17.37%). Compounds in C. reticulata peel oil consisted mostly of d‐limonene (62.39%) and γ‐terpinene (14.06%). The oils obtained from Z. zerumbet rhizome had α‐humulene (31.93%) and zerumbone (31.67%) as major components. The most abundant compounds in K. galanga rhizome oil were 2‐propeonic acid (35.54%), pentadecane (26.08%), and ethyl‐p‐methoxycinnamate (25.96%). The main component of S. aromaticum bud oil was eugenol (77.37%), with minor amounts of trans‐caryophyllene (13.66%). Assessment of larvicidal efficacy demonstrated that all essential oils were toxic against both pyrethroid‐susceptible and resistant Ae. aegypti laboratory strains at LC₅₀, LC₉₅, and LC₉₉ levels. In conclusion, we have documented the promising larvicidal potential of essential oils from edible herbs, which could be considered as a potentially alternative source for developing novel larvicides to be used in controlling vectors of mosquito‐borne disease.     

Larvicidal and mosquito repellent action of peppermint (Mentha piperita) oil

Oil of Mentha piperita L. (Peppermint oil), a widely used essential oil, was evaluated for larvicidal activity against different mosquito species: Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus by exposing IIIrd instar larvae of mosquitoes in enamel trays 6 × 4 inch² size filled to a depth of 3 inch with water. Of the three species tested Cx. quinquefasciatus was most susceptible followed by Ae. aegypti and An. stephensi. Application of oil at 3 ml/m² of water surface area resulted in 100% mortality within 24 h for Cx. quinquefasciatus, 90% for Ae. aegypti and 85% for An. stephensi. For Ae. aegypti 100% mortality was achieved at 3 ml/m² in 48 h or 4 ml/m² in 24 h. For An. stephensi 100% mortality was observed at 4 ml/m² in 72 h. The emergence at 3 ml/m² was also inhibited to a great extent and the few adults which emerged did not ovipost even after taking a blood meal. The oil showed strong repellent action against adult mosquitoes when applied on human skin. Percent protection obtained against An. annularis, An. culicifacies, and Cx. quinquefasciatus was 100%, 92.3% and 84.5%, respectively. The repellent action of Mentha oil was comparable to that of Mylol oil consisting of dibutyl and dimethyl phthalates.     

Survey of entomopathogenic nematodes and associate bacteria in Thailand and their potential to control Aedes aegypti

Aedes aegypti is an insect vector that transmits several viruses affecting humans worldwide. Entomopathogenic nematodes (EPNs) and their symbiotic bacteria are organisms with the potential to control many insects. In this study, we did a survey aimed to identify EPNs and their symbiotic bacteria and evaluate the larvicidal activity of bacteria against Ae. aegypti. We collected 540 soil samples from 108 sites in Phitsanulok Province, lower northern Thailand. Baiting techniques and White traps were used to isolate EPNs from soil samples. By sequencing of 28S rDNA and internal transcribed spacer regions, 51 EPN isolates were identified as Steinernema surkhetense (35 isolates), Heterorhabditis indica (14 isolates) and Heterorhabditis sp. SGmg3 (two isolates). Based on sequencing of a partial region of the recA gene, 35 isolates of Xenorhabdus were identified as Xenorhabdus stockiae, and 20 Photorhabdus isolates were identified as Photorhabdus luminescens subsp. akhurstii (10 isolates), P. luminescens subsp. hainanensis (seven isolates) and P. asymbiotica subsp. australis (three isolates). Screening for larvicidal activity of bacteria against Ae. aegypti was performed in the laboratory. Xenorhabdus WB5.4 and Xenorhabdus WB12.5, which were closely related to X. stockiae, resulted in high mortality of Ae. aegypti (99.99% and 70%, respectively) at 96 hr after exposure. Comparing with control groups, mortality of Ae. aegypti larvae was low (1.11%–6.67%) after exposure for 24–96 hr. Our findings showed the potential of X. stockiae for controlling Ae. aegypti. Further studies are needed to elucidate the mechanisms through which these bacteria kill Ae. aegypti larvae.     

Development of quinolone resistance and prevalence of different virulence genes among Shigella flexneri and Shigella dysenteriae in environmental water samples

The purpose of this study was to find out the mechanism of quinolone resistance in Shigella sp. isolated from environmental water samples from various parts of Kolkata, India. Out of 196 Shigella sp. isolated from 2014 to 2017, we selected 32 Shigella isolates for antimicrobial susceptibility tests. The minimum inhibitory concentrations (MIC) for quinolones ranged from 30 to 50 μg ml⁻¹ for ofloxacin, 5–20 μg ml⁻¹ for ciprofloxacin and 20–30 μg ml⁻¹ for norfloxacin. A few amino acid changes were found in quinolone resistance determining region (QRDR) of gyrA. Mutations in gyrA lead to a higher increment of MIC of quinolones. Among the plasmid-mediated (PMQR) quinolone resistance genes investigated, qnrB and aac(6')-lb-cr genes were found in all isolates. qnrA and qnrS were found in 25% and 62% of the isolates, respectively. ipaH gene was found in all of the isolates followed by the presence of other virulence genes ial, sen and stx1. Almost all the isolates having high MICs showed efflux pump activity in drug accumulation assay. All the mechanisms may or may not be present in a single strain. Several types of efflux pumps, presence of PMQR genes and mutations in drug target site of QRDR region may play the crucial role for resistance in our isolates.