Oil Of Betel Leaf Research Articles

Training on producing eucalyptus and betel leaf oil for dengue fever prevention

Dengue Hemorrhagic Fever (DHF) remains a significant public health issue in Indonesia and is endemic in nearly all provinces. The mortality rate from DHF continues to rise each year. To address this, health education on dengue fever prevention is essential. One preventive measure involves the use of natural ingredients. Betel leaves, in particular, can kill mosquito larvae due to the toxic substances they contain, which are especially harmful to mosquito larvae. Therefore, community service activities are necessary to raise awareness and provide practical solutions. This activity was conducted in Jetis Village, Sukoharjo, targeting 30 women in the Family Welfare Program (PKK) from the local community. The implementation included the preparation of pre-tests and post-tests, as well as training sessions. The questionnaire used in this activity assessed participants' skills through a pretest, followed by training in making eucalyptus and betel leaf oil, and concluded with a post-test. The results showed a significant improvement in the participants' skills, with most moving from "moderately competent" to "competent," as indicated by a P-value of 0.001. It can be concluded that the oil-making training effectively improved the skills of the PKK women.

Enhanced Disinfection Efficiency Using Cu Vortex Diode for Providing Safe Drinking Water: Devising Newer Methodologies

ABSTRACTThe present research aims to refine the hydrodynamic cavitation technique with traditional knowledgebase of Ayurveda for more efficient water disinfection and that has huge potential for implementation in real life, especially for substituting the existing chlorination method for drinking water treatment. The study incorporates use of newer copper reactor configuration, employing vortex flow for generating cavitation for the disinfection of water. Elimination of model contaminant, Escherichia coli, with initial concentration of ∼105 CFU/mL of bacteria was used for the disinfection study. Copper vortex diode with a capacity of 1 m3/h was employed as a cavitating device. The cavitation using copper vortex diode gave significantly higher disinfection, over 30%, compared that with conventional vortex diode, with aluminum as material of construction, under similar conditions. Remarkably, the addition of 0.1% betel leaf oil led to an extraordinary 260‐fold increase in the rate of disinfection, requiring only a single pass to achieve complete bacteria elimination. Furthermore, a notably high synergistic index of 246.96 was achieved for the process intensification approach using the Cu vortex diode. The cost was substantially reduced by approximately three times to 0.011 $/m3 using the Cu vortex diode compared to the conventional vortex diode. The developed strategy offers significantly enhanced performance, as well as a techno‐economical and sustainable solution for drinking water treatment to ensure the provision of safe drinking water. Moreover, the newer methodology can have the advantage of producing no harmful carcinogenic disinfection by‐products compared to chemical disinfection processes apart from sustainable alternative to chlorination.

Effect of ages on yield and quality of essential oil of betel leaf (Piper betle L.): Antioxidant activity, GC-MS and SEM analysis

In the present study, essential oil was extracted from the different ages (15, 30, 45 and 60 days) of betel leaves and evaluated their yield and quality of the essential oil. The effect of ages on percentage yield, chemical composition, morphological and physical properties, as well as antioxidant activity of essential oil was assessed and compared. The essential oil yield content of various ages of betel leaves, such as 15, 30, 45 and 60 days, is found to be 0.65%, 0.60%, 0.50% and 0.45%, respectively. Despite this, chemical compositions of extracted essential oil from various ages were analyzed by GC-MS analysis, and it was found that a total number of 55–65 volatile bioactive compounds are present, accounting for 97.78–98.88% of the extracted essential oil composition. The major bioactive compounds found in the all-ages of betel leaf essential oil are indanol (13.51–24.70%), elemene (4.36–10.84%) and eugenol (41.14%). Furthermore, 60 days old betel leaves essential oil exhibited higher antioxidant activity (87.01%) as compared to the 15 days younger betel leaves (84.05%). Moreover, SEM analysis was used to conduct a morphological investigation to examine the extraction mechanism of different stages of betel leaves. Therefore, obtained bioactive compounds from different ages of betel leaf essential oil can be used for various biological activities, food and pharmaceutical industries. In light of the findings, it is recommended that early-stage or younger leaves release higher essential oil while older leaves possess excellent antioxidant activity.

Effect of Ultrasonic Pretreatment on Yield and Properties of Essential Oil of Betel Leaf (Piper betle L.)

Effect of Ultrasonic Pretreatment on Yield and Properties of Essential Oil of Betel Leaf (Piper betle L.)

BETEL LEAF OIL AS A NATURAL DISINFECTION AGENT IN RADIOLOGICAL EQUIPMENT (STUDY OF MICROBIAL QUANTITY OF RADIOGRAPHIC CASSETTE)

Background: Betel leaf oil is a wonderful natural antiseptic. Betel leaf oil is rich in polyphenols, especially chavicol, which can be used to protect against microbe pathogens that cause infection. The cause of the spread of nosocomial infections in radiology can occur using a radiographic cassette. Previous research has found that radiographic cassette contains microorganisms and fungi. Purpose: To determine the effectiveness of using betel leaf oil as a natural disinfection on a radiographic cassette. Method: Quasi-experimental research with pre-test and post-test design designs was carried out by calculating the number of microbes, including Total Plate Count (TPC), Staphylococcus, and fungi. The study on 12 conventional radiographic cassettes was calculated using the swab method before and after cleaning with betel leaf oil. The data were analyzed descriptively to illustrate the rate of decline. Result: The results showed that the percentage rate of decline after the radiographic cassette was cleaned with betel leaf oil was ALT (17.8%), Staphylococcus (57.41%), and fungi (37.21%). Conclusion: Betel leaf oil can be used as alternative natural disinfection on radiographic cassettes, quite effectively suppressing the activity and number of microbes present.

Green steam for sustainable extraction of essential oils using solar steam generator: A techno-economic approach

Essential oils are natural volatiles extracted using steam. Steam generation is an energy-intensive process and is invariably based on fossil fuels, briquettes or biomass. Based on the volumes of essential oil extracted, it can be considered as significant contributor to greenhouse gases. Though a lot of work is reported on essential oil extraction and characterisation, method for quantification of exact steam requirement and means for achieving it through sustainable source is not much reported.Through this study, steam requirement of 750 g/ml of Betel leaf volatile oil extracted and quantification of energy requirements for the extraction process is determined. For adoption of sustainable and clean energy source, opto-geometric optimisation and concentrating solar collectors (CSC) area calculation are carried out. The design of three CSC, namely, Linear Fresnel Reflector (LFR), parabolic dish collector (PDC) and parabolic trough collector (PTC), resulted in 78 m2, 122 m2 and 138.87 m2 of area to meet the steam requirement of the process.Adoption of CSC technologies for steam generation and replacing the prevalent technology needs detailed analysis and techno-economic comparison. This paper also focuses on cost-benefit analysis, taking into account CAPEX, OPEX, life-span, return on investment, levelized energy (steam) cost and the profitability index of net present value, payback period and internal rate of returns to check the economic feasibility of renewable energy. Analysis indicates that the cost associated with unit steam generation through CSC over the specified period is lower than a briquette-based steam generation. On adopting this sustainable technology for betel leaf oil extraction, even if the initial cost seems to be high, the payback period of LFR and PTC are 1.10 and 2.45 years, respectively. Based on this study, CSCs can provide a sustainable green solution for the extraction of essential oils.

Karakteristik Mikroemulsi Minyak Daun Sirih (Piper betle L.) pada Perlakuan Rasio Campuran Surfaktan dan Minyak Daun Sirih

This study aims to determine the effect of the mixture ratio of surfactant and betel leaf oil on the characteristics of the microemulsion of betel leaf oil and determine the ratio of the mixture of surfactant and betel leaf oil that is appropriate to obtain the best microemulsion characteristics. This experiment used a Randomized Block Design (RAK), using the surfactant: betel leaf oil ratio treatment with 10 levels, namely 100:0; 98:2; 96:4; 94:6; 92:8; 90:10; 88:12; 86:14; 84:16 and 82:18. Grouping based on processing time. The best microemulsion is the microemulsion which has the highest concentration of betel leaf oil. The best microemulsion was tested for stability against pH and dilution during 8 weeks of storage. The turbidity index value is tested every 2 weeks. The rate of damage was calculated using linear regression analysis. The microemulsion was made by mixing the surfactant and betel leaf oil (5 ml in the oil phase), then adding the demineralized distilled water (water phase) dropwise in volume (10 ml). The microemulsion was incubated for 24 hours and calibrated using a spectrophotometer at a wavelength of 502 nm. The results showed that the ratio of the mixture of surfactants and betel leaf oil had an effect on the characteristics of the microemulsion of betel leaf oil. The ratio of surfactant and betel oil 94:6 is the best treatment for making betel leaf oil microemulsion with microemulsion characteristics that have a transparent appearance, turbidity index values ??before and after centrifugation are 0.225±0.005% and 0.196±0.005% and have a particle size of 14.8 ±6.9 nm, and the largest droplet size was 11 nm. Betel leaf oil microemulsion with surfactant and betel leaf oil ratio of 94:6 was stable for 8 weeks of storage.
 Keywords: microemulsion, ratio, surfactant, Piper betle Linn.

In vitro efficacy of essential oils against Colletotrichum gloeosporioides, the causal agent of Anthracnose

Colletotrichum gloeosporioides Penz, the anthracnose disease causing fungal pathogen account for upto 50% spoilage of fresh fruits and vegetables. The present study was conducted to explore anon-chemical approach for controlling the postharvest anthracnose disease.Antifungal activities of essential oils from thyme (Thymus vulgaris L), betel leaf (Piper betle), cinnamon (Cinnamomum verum) and oregano (Origanum vulgare) were determined against C. gloeosporioides through in vitro poisoned food technique. Partial or complete inhibition of the pathogen was recorded by use of oils from thyme, betel leaf, cinnamon, and oregano at 100400 ppm levels. In terms of the EC50 values, the performance of antifungal activity of cinnamon oil was highest followed by oregano, thyme and betel leaf. Characterization through GC-MS yielded the major constituents from thyme oil as thymol, cymene; caryophyllene in betel leaf oil; trans-cinnamaldehyde in cinnamon oil and carvacrol in oregano oil. The present findings suggest that natural plant based essential oils can emerge as promising agents for preventing spoilage of horticultural commodities from anthracnose disease.

Electrospun Essential oil encapsulated nanofibers for the management of anthracnose disease in Sapota

Control of anthracnose disease caused by Colletotrichum gloeosporioides by use of essential oils encapsulated in nanofibres made of polyvinyl alcohol has been attempted in this study. Essential oils from plants of subtropical origin, thyme oil and betel leaf oil were incorporated in electrospun mats at different levels. The Scanning Electron Microscope analysis revealed size of the nanofibers between 180 and 302 nm. Effective Dose (ED50) values for the pure oils and their blends (1:3; 1:1; 3:1) ranged from 77 to 148 ppm. The blended essential oils demonstrated additive effect for inhibition of the pathogen with Abbott index >0.5. Oil loading capacity, determined using Gas Chromatography Electron Capture Detector ranged from 40.15–75.88% and was found proportional to feed content. No major effect of incorporation of thyme oil and betel oil on polymeric structure was observed in Fourier-transform infrared spectra. A Band near 876cm−1 and 1250 cm−1 signified the presence of both thyme and betel oil respectively. The composite nanofibers with blended oils at three levels showed inhibition zone from 6.6 mm to 11.6 mm on 3rdday of incubation, which increased further upto 24 mm on the 6th day. Sapota fruits inoculated with Colletotrichum gloeosporioidesrecorded disease severity of 83.33 %, 8.39 % and 6.72 % when packed in corrugated fiber board boxes with composite nanofibers as compared to control fruits (100 %) on the 6th day of incubation at 28 °C. Burning symptoms on skin were observed on fruits kept in contact with same level of essential oil infused sterile cotton balls highlighting the importance of encapsulated oils for disease control.

Formulation and characterization of betel leaf (Piper betleL.) essential oil based nanoemulsion and its in vitro antibacterial efficacy against selected food pathogens

The current study is focused on the formulation and characterization of oil-in-water nanoemulsion of betel leaf (Piper betle L.) essential oil and its antibacterial efficacy. The nanoemulsion consists of essential oil of betel leaf as the dispersed phase, distilled water as a continuous phase and a nonionic surfactant Tween 20 (Polysorbate 20) as an emulsifier. Six emulsions were formulated varying the oil to surfactant ratio (vol/vol) from 1:1 to 1:6, using high-energy emulsification process. Three samples, that is, S2 = 1:2 (3% EO, 6% Tween 20), S3 = 1:3 (3% EO, 9% Tween 20), and S4 = 1:4 (3% EO, 12% Tween 20) shown droplet diameter of 50 ± 8, 70 ± 12, and 175 ± 7 nm, respectively. The formulated nanoemulsions showed antimicrobial activity as minimum inhibitory concentration (MIC) of 0.5–1.25 µl/ml and minimum bactericidal concentration (MBC) of 1–2.5 µl/ml against five strains of gram-positive and gram-negative bacteria. The formulated nanoemulsions thus can serve as natural antimicrobial agent for food system. Practical application This study can be useful in understanding the potential of essential oil from plant sources in food processing. It can also be helpful to formulate nanoemulsion from essential oil and its characterization. From characterization, researchers can get the knowledge about changes in the morphological characteristics of the nanoemulsion and also other physical properties. The thermodynamic and kinetic stability study of the nanoemulsions can give us an idea about the application in different food processing and preservation conditions. Furthermore, the use of essential oil in the food industry is mainly as a flavouring agent. Based on the antibacterial efficacy of the formulated nanoemulsions, it can also be utilized as a potential natural class one preservative.

Acute toxicity of chemical pesticides and plant-derived essential oil on the behavior and development of earthworms, Eudrilus eugeniae (Kinberg) and Eisenia fetida (Savigny).

Comparative toxicity of two chemical pesticides (temephos and monocrotophos) versus a plant-derived betel leaf oil Piper betle (L.) to earthworm Eudrilus eugeniae (Kinberg) and redworm Eisenia fetida Savigny, historically: Eisenia foetida (Savigny 1826), was evaluated. Mortality rate was more prominent in temephos at 100μg concentration to both the earthworms in filter paper test (FPT) as well as 10mg concentration in artificial soil test (AST). In contrast, P. betle does not display much mortality rate to both the earthworms even at 1000mg of treatment concentrations. The lethal concentration (LC50) value was observed at 3.89 and 5.26mg/kg for temephos and monocrotophos against E. eugeniae and 3.81 and 5.25mg/kg to E. fetida, respectively. Whereas, LC50 value of betel leaf oil was only observed at 3149 and 4081mg/kg to E. eugeniae and E. fetida, respectively. Correspondingly, the avoidance or attraction assay also displayed that earthworms were more sensitive to the soil containing chemical pesticides. Whereas, the avoidance percentage was decreased in the P. betle oil. Similarly, sublethal concentration of chemical pesticides (5 and 6.5mg) significantly reduced the earthworm weight and growth rate. However, P. betle oil did not change the developmental rate in the duration of the assay (2, 7 and 14days) even at 4000mg treatment concentration. The enzyme ratio of CAT and SOD was also affected significantly after exposure to the chemical pesticides (6.5mg/kg). Hence, our study implied the risk assessment associated with the chemical pesticides and also recommends plant-derived harmless P. betle oil against beneficial species as an alternative pest control agent.

Comparative Evaluation of Antifungal Activity of Piper Betel Leaf Oil, Origanum vulgare Essential Oil and Fluconazole Suspension on Candida albicans − An In Vitro Study

Introduction: Oropharyngeal candidiasis is an opportunistic mucosal infection caused by Candida albicans. It usually responds to topical treatments such as clotrimazole troches, topical fluconazole, chlorhexidine mouthwash and nystatin suspension. Piper betel leaf oil and Origanum vulgare essential oil have shown some topical antifungal activity. Aim: To determine and compare the antifungal efficacy of piper betel leaf oil, O. vulgare essential oil and fluconazole suspension against C. albicans. Materials and Methods: The zone of inhibition was measured by the cup–plate diffusion method using 100 μl volume of piper betel leaf oil, O. vulgare essential oil and fluconazole suspension, which were pipetted into the wells of the inoculated Sabouraud’s dextrose agar plates. The zone of inhibition was measured in millimetres using Vernier calliper. Minimum inhibitory concentration (MIC) was determined by the broth macro-dilution test by pouring 1 ml of the respective concentrations of the test material to the individual test tubes along with 10 μl of the diluted test organism inoculum. Finally, MIC was calculated using a reflective viewer. Results: The zone of inhibition for O. vulgare essential oil (>40 mm) was more than fluconazole suspension (>35 mm). MIC of O. vulgare essential oil, piper betel leaf oil and fluconazole suspension was 1.6%, 0.4% and 0.8%, respectively. Conclusion: O. vulgare essential oil was found to be a more effective antifungal agent than piper betel leaf oil and fluconazole suspension.

In vitro evaluation of the effects of some plant essential oils on Ascosphaera apis, the causative agent of Chalkbrood disease.

Ascosphaera apis is one of the major fungal pathogens of honey bee broods and the causative agent of Chalkbrood disease. The factors responsible for the pathogenesis of Chalkbrood disease are still not fully understood, and the increasing resistance of A. apis to commonly used antifungal agents necessitates a search for new agents to control this disease. The in vitro antifungal activities of 27 plant essential oils against two isolates of A. apis (Aksu-4 and Aksu-9) were evaluated. Out of the 27 plant essential oils tested, 21 were found to be effective in killing both isolates of A. apis. Based on their minimum fungicidal concentration (MFC) values, the effective oils were grouped into three categories: highly effective, moderately effective and minimally effective. Mountain pepper oil, Kala Bhangra oil, spearmint oil, babuna oil, betel leaf oil, carrot seed oil, cumin seed oil and clove bud oil were highly effective, with MBC values between 50.0 μg/mL and 600.0 μg/mL. Mountain pepper was the most effective essential oil, with an MBC value of 50.0 μg/mL. Citral and caryophyllene containing oils were the most effective with MIC 50 ppm. The essential oils tested exhibited significant antimicrobial activities against both strains of A. apis, and they may contain compounds that could play an important role in the treatment or prevention of Chalkbrood disease of honeybee.

Developmental response of Spodoptera litura Fab. to treatments of crude volatile oil from Piper betle L. and evaluation of toxicity to earthworm, Eudrilus eugeniae Kinb.

Evaluations of biological effects of (Pb-CVO) the crude volatile oil of Piper betle leaves on the tobacco cutworm Spodoptera litura were conducted. Pb-CVO was subjected to GC-MS analysis and twenty vital compounds were isolated from the betel leaf oil. Pb-CVO was tested at four different concentrations (0.25, 0.5, 1.0 and 1.5%) against S. litura. The treated insects exhibited dose depended mortality. The mortality rate was significantly higher at the 1.0 and 1.5% Pb-CVO. The LC50 (Lethal concentration) were observed at 0.48% Pb-CVO. Larval and pupal durations increased in all treatment concentrations (0.25, 0.3, 0.4 and 0.5%) whereas, pupal weight decreased compared to control. Adult longevity of S. litura was reduced in all treatments but predominantly in the 0.4 and 0.5% Pb-CVO. Correspondingly, mean fecundity rate was reduced at all concentrations compared to control. Histological studies of larvae mid-gut profiles of S. litura were severely damaged in 1.0 and 1.5% and showed abnormalities in mid-gut cells with 0.25 and 0.5% Pb-CVO treatments. Earthworm toxicity illustrated that 0.1% of chemical insecticides (monocrotophos and cypermethrin) varied widely in their contact toxicities compared to 0.5 and 1.0% Pb-CVO and control in both contact filter paper and artificial soil test. These findings suggest that twenty essential compounds of betel leaf oil were significant inhibitors of the development and caused behavioral changes of S. litura. Treatment with betel leaf oil at these concentrations had no adverse effect on earthworm populations.

Modelling the effect of essential oil of betel leaf (Piper betle L.) on germination, growth, and apparent lag time of Penicillium expansum on semi-synthetic media

The current study aimed at characterizing the chemical components of betel leaf (Piper betle L. var. Tamluk Mitha) essential oil (BLEO) and modelling its effect on growth of Penicillium expansum on semi-synthetic medium. Gas chromatography-mass spectrophotometry (GC-MS) analysis of BLEO revealed the presence of different bioactive phenolic compounds in significant amounts. Among 46 different components identified, chavibetol (22.0%), estragole (15.8%), β-cubebene (13.6%), chavicol (11.8%), and caryophyllene (11.3%) were found to be the major compounds of BLEO. A disc diffusion and disc volatilization method were used to evaluate antifungal activity of the oil against a selected food spoilage mould. The logistic model was used to study the kinetics of spore germination. Prediction and validation of antifungal effect of BLEO was performed on semi-synthetic medium (apple juice agar) using predictive microbiological tools. The Baranyi and Roberts model was used to estimate maximum growth rate (μmax in mm/day) and apparent lag time (λ in days) of the mould. Secondary modelling was performed using a re-parameterized Monod-type equation based on cardinal values to study the effect of different BLEO concentration on estimated growth parameters. Emax (minimum concentration of oil at which mould growth was inhibited) and MIC (minimum inhibitory concentration of BLEO at which lag time is infinite) value of BLEO against P. expansum was estimated to be 0.56 and 0.74μl/ml, respectively, which was found to be similar on potato dextrose agar (PDA) as well as apple juice agar (AJA) medium. The correlation between estimated growth parameters of the mould on both the media was obtained with satisfactory statistical indices (R2 and RMSE). This study revealed inhibitory efficacy of BLEO on spore germination, mycelial growth and apparent lag time of P. expansum in a dose-dependent manner. Hence, BLEO has potential to be used as a natural food preservative.

Development of a novel cup cake with unique properties of essential oil of betel leaf (Piper betle L.) for sustainable entrepreneurship.

Betel vine (Piper betle L.) is a root climber with deep green heart shaped leaves. It belongs to the Piperaceae family. There is a huge wastage of the leaves during glut season and it can be reduced by various means including extraction of medicinal essential oil which can be considered as GRAS (generally recognized as safe) materials. Therefore, attempts were made to develop a novel cup cake by incorporating essential oil of betel leaf. The textural properties of the cakes were measured by texture analyzer instrument; whereas the organoleptic properties were adjudged by human preferences using sensory tables containing 9-point hedonic scale. Price estimation was done considering all costs and charges. Finally, all parameters of the developed cake were compared with different cup cakes available in the market for ascertaining consumer acceptability of the newly developed product in terms of quality and market price. Results revealed that the Novel cup cake developed with 0.005% (v/w) essential oil of betel leaf occupied the 1st place among the four developed novel cup cakes. However, it occupied 4th place among the nine cup cakes in the overall preference list prepared based on the textural and organoleptic qualities, though its market price was calculated to be comparable to all the leading cupcakes available in the market. This indicates that manufacturing of novel cup cake with essential oil of betel leaf would be a profitable and self-sustaining entrepreneurship.

Fungal liver abscess in an immunocompetent individual

1. Gupta PJ. Human myiasis in anal carcinomatous ulcer – a case report. Eur Rev Med Pharmacol Sci. 2009;13:473–4. 2. Gupta PJ. Myiasis: a perianal wound infection caused by fly larvae in gangrenous hemorrhoid. Tech Coloproctol . 2006;10:254–5. 3. da Silva AL, Santos Ide O. Ano-rectal myiasis. Rev Bras Med. 1965;22:273–5. 4. Calzaretto J, Fuenzalida E, Litrenta SR. Anal condyloma acuminata with myiasis. Prensa Med Argent. 1965;52:2282–3. 5. Lukin LG. Human cutaneous myiasis in Brisbane: a prospective study. Med J Aust. 1989;150:237–40. 6. Hope FW. On insects and their larvae occasionally found in the human body. Trans R Soc Entomol. 1840;2:256–71. 7. Zumpt F. Myiasis in Man and Animals in the Old World. London: Butterworths, 1965.p.89–102, 267. 8. Sherman RA, Hall MJ, Thomas S. Medicinal maggots: an ancient remedy for some contemporary afflictions. Annu Rev Entomol. 2000;45:55–81. 9. Kumarasinghe SP, Karunaweera ND, Ihalamulla RL, Arambewela LS, Dissanayake RD. Larvicidal effects of mineral turpentine, low aromatic white spirits, aqueous extracts of Cassia alata, and aqueous extracts, ethanolic extracts and essential oil of betel leaf (Piper betle) on Chrysomya megacephala. Int J Dermatol. 2002;41:877–80. 10. Kumarasinghe SP, Karunaweera ND, Ihalamulla RL. A study of cutaneous myiasis in Sri Lanka. Int J Dermatol. 2000;39:689–94.

Larvicidal efficacy of essential oil of betel leaf ( Piper betle ) on the larvae of the old World screwworm fly, Chrysomya bezziana in vitro

Larvae of <i> Chrysomya bezziana</i> are the commonest cause of wound myiasis in some parts of the world. This obligatory parasite is important in humans and in commercial livestock. Kumarasinghe <i> et al </i> have reported that essential oil of betel leaf (EOBL) is larvicidal to <i> C. megacephala </i> but there are no publications on its effect on <i> C. bezziana</i> . This study was done to evaluate the efficacy of essential oil of betel leaf (<i> Piper betle</i> ) against the larvae of <i> C. bezziana</i> <i> in vitro</i> . EOBL was prepared at the Industrial Technology Institute Colombo, Sri Lanka, according to a standard protocol. The experiment on larvae was carried out at the Research Institute for Veterinary Sciences in Bogor, Indonesia. EOBL concentrations of 2%, 3% and 4% were prepared with Tween 80. Two ml of 4%, 3% and 2% EOBL in 1% Tween 80 (v/v/aq) were poured into separate Petri dishes. Ten 1st and 2nd instar larvae were placed in each Petri dish. Asuntol (Chaumaphos) 1% was used as positive control and distilled water with 1% tween 80 was the negative control. Larval mortality was assessed half-hourly. The experiment was repeated five times and averages were compared. Sustained immobility of the larvae, after exposure to the relevant substances was considered as death. The efficacy of EOBL depended on, the stages of <i> C. bezziana</i> larvae and the concentration. With 4% EOBL, all first instar larvae were killed within two hours and the second instar larvae were killed by four hours. The positive control showed no mortality until four hours but all larvae were weak, from the first 30 minutes. In the negative control, larvae were mobile and active. EOBL 3% killed all the first instar larvae by 150 minutes and 74% of the second instar at four hours. By 210 minutes, 2% preparation had killed 100% of the first instars. EOBL is an effective larvicidal for <i> C. bezziana</i> first and<i> </i> second instar larvae <i> in vitro</i> . This natural product has a great potential to be developed as a novel larvicide against this parasite.

Larvicidal effects of mineral turpentine, low aromatic white spirits, aqueous extracts of Cassia alata, and aqueous extracts, ethanolic extracts and essential oil of betel leaf (Piper betle) on Chrysomya megacephala.

Many methods have been employed, with variable success, in the treatment of cutaneous myiasis caused by Chrysomya species. Experiment 1: to assess the larvicidal effect of mineral turpentine (MT) and the main ingredient of MT, low aromatic white spirits (LAWS), on Chrysomya megacephala larvae in vitro. Experiment 2: to assess the larvicidal effects of aqueous extracts of winged senna (Cassia alata), and aqueous extracts, ethanolic extracts and essential oil of betel leaf (Piper betle). In experiment 1, two samples of LAWS were obtained from two industrialists (samples 1 and 2). Adult flies of C. megacephala were bred in the insectory of the Department of Parasitology, Faculty of Medicine, University of Colombo. Petri dishes were prepared with pads of cotton wool. These cotton pads were soaked separately in MT, LAWS samples 1 and 2, and normal saline as a control. Ten larvae were placed in each Petri dish. The activity of the larvae was observed and recorded half-hourly. MT and the two samples of LAWS were analyzed by chromatography. In experiment 2, volatile essential oil of betel was prepared using a standard steam distillation process. An ethanolic extract of betel was obtained after boiling the crushed leaf with water, and mixing the stock with ethanol. Betel oil dilutions of 1-4% were prepared using 1% Tween 80 (v/v aq) as a solvent, with 0.05 g/100 mL sodium lauryl sulphate (as stabilizer) and 0.01 g/100 mL methyl paraben (as a preservative). Cotton wool swabs soaked in 1, 2, 3 and 4% essential oil of betel in 1% Tween 80 (v/v aq) prepared as above, 1, 2, 3 and 4% ethanolic extract of betel, 50 and 25% aqueous extract of C. alata, and 50 and 25% aqueous extract of betel were placed in separate Petri dishes. Ten larvae were placed in each Petri dish. 1% Tween 80 solvent with the stabilizer and the preservative, but without betel essential oil, was used as a negative control and MT was used as a positive control. Larval motility was assessed as before. MT and the two LAWS samples killed the larvae in vitro within 4 h. Chromatography showed more unidentified constituents in MT than in pure LAWS, indicating additional substances in MT. The 4 and 3% preparations of the essential oil of betel were effective in killing 100% of the larvae of Chrysomya within 3 h 30 min. The 2% extract of betel essential oil killed 96.7% of larvae in 4 h. Ethanolic and aqueous extracts of betel, the aqueous extract of C. alata, normal saline and the Tween 80 solvent were not larvicidal. MT and LAWS, the main ingredient of MT, were effective in killing Chrysomya larvae. Essential oil obtained from betel leaves also showed a dose-dependent larvicidal effect on Chrysomya larvae. This natural product may be effective in the treatment of wound myiasis.