Premna cerealella - Hindawi Publishing Corporationdownloads.hindawi.com/archive/2015/319045.pdfResearch Article Chemical Composition and Insecticidal and Repellent Effect of Essential

Research ArticleChemical Composition and Insecticidal and Repellent Effect ofEssential Oils of Two Premna Species against Sitotrogacerealella

Elvis Adjalian12 Philippe Sessou1 Theacuteophile Odjo2 Gilles Figueredo3 Dansou Kossou2

Feacutelicien Avlessi1 Chantal Menut4 and Dominique Sohounhloueacute1

1Laboratoire drsquoEtude et de Recherche en Chimie Appliquee (LERCA) Ecole Polytechnique drsquoAbomey-CalaviUniversitedrsquoAbomey-Calavi 01 BP 2009 Cotonou Benin2Laboratoire de Production Vegetale Faculte des Sciences AgronomiquesUniversite drsquoAbomey-Calavi 01 BP 526 Cotonou Benin3Laboratoire drsquoAnalyse des Extraits Vegetaux et des Aromes (LEXVA Analytique) 460 rue du Montant 63110 Beaumont Benin4Institut des Biomolecules Max Mousseron Equipe ldquoGlyco et Nanovecteurs pour le Ciblage Therapeutiquerdquo Faculte de Pharmacie15 avenue Charles Flahault BP 14491 34093 Montpellier France

Correspondence should be addressed to Dominique Sohounhloue ksohounbjreferorg

Received 2 March 2015 Revised 26 June 2015 Accepted 8 July 2015

Academic Editor Fedai Erler

Copyright copy 2015 Elvis Adjalian et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

This work aims to study for the first time the chemical composition and evaluate insecticidal and repellent effects of essentialoils of Premna angolensis and Premna quadrifolia leaves against Sitotroga cerealella an insect pest of rice stocks as alternatives tosynthetic pesticidesTheGC-MS analysis showed that essential oil of P angolensis contains 29 compounds representing 961 of theoil and 42 compounds corresponding to 91 for the essential oil of P quadrifolia The main constituents regardless of the specieswere 120573-caryophyllene (131) (E)-120573-caryophyllene (135) octen-3-ol (32ndash28) phytol (37ndash49) 120573-elemene (14ndash21)globulol (112) germacrene-D (89) 120572-humulene (29ndash64) 120572-pinene (5) sabinene (37) 120575-cadinene (04ndash33) andlinalool (33) The results of laboratory tests showed that both essential oils have insecticidal and repellent effects on S cerealellaPresenting the results the damage caused by the adults and larvae of S cerealella was evaluated by calculating the percentage ofgrains attacked and weight loss thereof The results suggest that volatile extracts of P angolensis and P quadrifolia can be used asalternatives to synthetic chemicals in paddy protection against S cerealella

1 Introduction

The genus Premna L (Verbenaceae) contains about 200species worldwide which are distributed mainly in tropicaland subtropical parts ofAsia Africa Australia and the PacificIslands [1 2] Previous studies on chemical constituents of thegenus Premna led to the isolation of triterpenes diterpenesand sesquiterpenes [3] some of them exhibiting significantantibacterial activities [4] When antiparasitic activity of newCaledonian medicinal plants including Premna serratifolia Lwas evaluated it was observed that P serratifolia was activeagainst Leishmania donovani with IC

50values between 05

and 5 120583gsdotmLminus1 [5] In addition some preparations of plantsof the genus Premna have been used in the treatment of

liver disorders and antioxidant as well as for their immunemodulatory effects [6] in the traditional Indian system ofmedicine The Premna genus can be used in treating variousailments like rheumatism asthma dropsy cough fever boilsand scrofulous disease Traditionally in tropical Africa andBenin in particular people with low purchasing power veryoften use odors produced by aromatic plants collected locallyfor the control of insectsThis is the case of Premna angolensisGurke and Premna quadrifolia Schum amp Thonn leaves ofwhich are burned and used by producers as a fumigant inthe attics of cereals against pests In this study extracts fromleaves of both species P angolensis and P quadrifolia har-vested in Benin were analyzed by gas chromatography-massspectrometry (GCMS) for the first time The insecticidal

Hindawi Publishing CorporationJournal of InsectsVolume 2015 Article ID 319045 6 pageshttpdxdoiorg1011552015319045

2 Journal of Insects

and repellent activities were tested on the Angoumois grainmoth Sitotroga cerealella (L) (Lepidoptera Gelechiidae) inrice storage

2 Material and Methods

21 PlantMaterial andDistillation of the Volatile ConstituentsLeaves of P angolensis and P quadrifolia were collectedfrom the municipality of Come by the authors and certifiedat the National Herbarium of Abomey-Calavi UniversityThe collected plant materials were stored in the laboratorybetween 18 and 20∘C in the shade of the sunlight throughoutthe period of extraction The essential oils were obtained byhydrodistillation of the leaves (450 g) for five hours usinga Clevenger-type extractor according to British Pharma-copoeia method [7] They were then dried over anhydroussodium sulfate and analyzed by GCMS

22 Test Organisms Individuals of S cerealella used formass rearing for this study come from the reserve WARDA(Benin) Rearing conditionswere119879= 29plusmn2∘C RH= 70plusmn10

23 Analysis of the Volatile Constituents

231 GCMS The essential oils were analysed on a Hewlett-Packard gas chromatograph Model 7890 coupled to aHewlett-Packard MS Model 5875 equipped with a DB5 MScolumn (30mtimes 025mm 025 120583m) programming from 50∘C(5min) to 300∘C at 5∘Cmin 5min hold Helium was used ascarrier gas (10mLminminus1) injection was made in split mode(1 30) injector and detector of temperature were at 250 and280∘C respectivelyTheMSworking in electron impactmodewas at 70 eV electron multiplier was 2500 eV ion sourcetemperature was 180∘C mass spectra data were acquired inthe scan mode inmz range 33ndash450

232 GCFID Theessential oils were analyzed on aHewlett-Packard gas chromatograph Model 6890 equipped with aDB5 MS column (30m times 025mm 025 120583m) programmingfrom 50∘C (5min) to 300∘C at 5∘Cmin 5min hold Hydro-gen was used as carrier gas (10mLminminus1) injection was insplit mode (1 60) injector and detector temperature were280 and 300∘C respectively The essential oil was diluted inhexane 130 The compounds assayed by GC in the differentessential oils were identified by comparing their retentionindices with those of reference compounds in the literatureand confirmed by GC-MS by comparison of their massspectra with those of reference substances [8ndash10]

24 Test All tests were performed at a temperature of 29plusmn2∘Cand natural photoperiod with relative humidity 70 plusmn 10

25 Contact Toxicity Tests with Essential Oils Bioassays wereperformed using the method described by Noudogbessi et al[11] The temperature of the test medium ranged from 25∘Cto 31∘C and relative humidity was 80 Five concentrations(0 3 5 10 and 15 120583LmLminus1) depending on the selected

75 cm

4 cm

Figure 1 Olfactometer set-up An individual female Angoumoisgrain moth was introduced in the center of the tube Her positionwas recorded after 15min since introduction Treated and controlpaddy rice were positioned at either end of the tube

chemical profile of each essential oil in absolute ethanolsolution were tested The plant material paddy rice grainsof the variety ldquoIR841rdquo was treated with each essential oilAfter 24 h 5 males and 10 females of S cerealella aged 0 to24 h were deposited on the treated plant material Overall5 doses times 3 repeats times 2 types of oil = 30 experimental units(containers) were implemented The control sample treatedonly with ethanol was also infested by S cerealella Adultmortality was monitored 96 h after exposure to the essentialoils Then the insects were separated from the grains Theemergence of new insects was then observed at intervals of24 h in the experimental units to the 50th day after infestationAssessment concerned the rate of infested grains and weightloss of grains of rice

To assess the S cerealella damage on the rice seeds aftertreatment the substrate grains were collected counted andweighed The rate of weight loss of dry material of the grainswas determined according to MCP (Method of CountingandWeighing) andMSVW (Method of Standard VolumetricWeight) [12] Two criteria of assessment of the damage arecommonly used the percentage of attack of grains (119860) andpercentage of weight loss (119861) [13] respectively calculatedby the equations

119860 = NaNa +Ns

times 100

119861 = PsNa minus PaNsPs (Na +Ns)

times 100

(1)

where Na is number of attacked grains Ns is number ofhealthy grains Pa is weight of damaged grains and Ps isweight of healthy grains

26 Repellency Tests The repellent action of the plants wastested in an olfactometer (Figure 1) consisting of a 75 cmglasstube of 4 cm in diameter with a 29 mm hole in the middleAt either end of the tube a small jar was placed containingeither 100 g containing paddy rice treated with only ethanolor 100 g of paddy rice seedsmixed with a solution of essentialoil at concentrations 01 05 and 1 The hole in themiddle was covered with gauze whereas the ends of the tubewere closed by putting a plastic petri dish against them Airwas gently (ca I mls) sucked away from the centre of the tubeto prevent the accumulation of plant odor in the tube

Ten female S cerealella aged 24 h to 48 h were releasedone after the other in themiddle of the tube through the holeInsect behavior was observed and the position was recordedfor 15min Both volatile extracts were tested once before

Journal of Insects 3

the second round Insects were considered to have made achoice when entered in one of the jars containing rice grainsor those who have reached the last twenty-five centimeters ofthe tube For those who chose the middle of the tube theyhave finally made a choiceThe percentage of insects rejectedwas calculated using the following formula

Percentage Repellency () = 119860 minus 119861119860 + 119861times 100 (2)

where119860 is average number of insects present in the untreatedportion (insects repelled) and 119861 is average number of insectsin the treated (not repelled insects) part

The average percentage of repulsion for the essential oilwas calculated and assigned according to previous ranking[14] to one of several repulsive classes ranging from 0 to Vclass 0 (Percentage Repellency (PR) lt 01) class I (PR = 01ndash20) class II (PR=201ndash40) class III (PR=401ndash60) classIV (PR = 601ndash80) and class V (PR = 801ndash100)

27 Statistical Analysis The results from the observationswere statistically processed byAnalysis ofVariance (ANOVA)using SAS (Statistical Analysis System) Version 91 Finallyan average of the percentages of the three results of the testwas made The results of statistical tests were consideredsignificantly different if the probability of the hypothesis wasless than or equal to 5

3 Results

Chromatographic analysis of the essential oil extracted fromleaves of P angolensis detected 29 compounds representing961 of the weight of the essential oil (Table 1) Howeversesquiterpenes hydrogenated (266) and oxygen (20)werepredominant As for P quadrifolia 43 compounds were iden-tified representing 91 of the essential oil It was dominatedby the hydrocarbon sesquiterpenes (655) (Table 1) Thepredominant compounds of the two essential oils regardlessof species and their origin were 120573-caryophyllene (131)(E)-120573-caryophyllene (135) octen-3-ol (32ndash28) phy-tol (37ndash49) 120573-elemene (14ndash21) globulol (112)germacrene-D (89) 120572-humulene (29ndash64) 120572-pinene(5) sabinene (37) 120575-cadinene (04ndash33) and Linalool(33)

The results of the estimation of the influence of differentconcentrations (0 to 15120583LmLminus1) of the two essential oilson adult mortality of S cerealella the emergence of younginsects the rate of grains attacked and loss correspondingweights are shown in Tables 2 and 3 Mortality rates of insectsin Table 2 were obtained 4 days after treatment of the grainsof paddy rice which ranged between 0 and concentrations15 120583LmLminus1 At a concentration of 3 120583LsdotmLminus1 P angolensisessential oil showed over 90 mortality through its contactactivity on the adults of S cerealella The number of grainsdamaged and the loss of the weight of rice decrease whenthe dose of essential oil increases The insecticidal effect ofP quadrifolia essential oil was less noticed compared to theessential oil of P angolensis with significant difference (119875 lt0001) In the controls sample we noticed that the number of

adults of S cerealella increased rapidly and reached the rateof 8962 plusmn 001 with P quadrifolia and 8571 plusmn 003 with Pangolensis

Repellent Effect of Essential Oils Tested The percentages ofrepulsion for the different doses of the essential oil of theleaves of P quadrifolia and P angolensis are summarized inTable 4 The different doses of essential oils (01 05 and1) caused 6711 to 9684 repellency for P angolensis and6555 to 9155 for P quadrifoliaThis clearly shows that thepercentage of repulsion increases with the dose

4 Discussion

The output value of the essential oil from the leaves of Pquadrifoliawas relatively better (0102 plusmn 0005) than that ofP angolensis (0056 plusmn 0004) No data have been reportedin the literature on the essential oils of those two speciesEssential oils P quadrifolia and P angolensis showed dif-ferent chemical compositions They were potentially rich insesquiterpenoid compounds During this study insecticidaland repellent activity of both oils against S cerealella wasvalued according to the treatment and the dose of appliedoil Previous studies on other species of Premna showedthat their chemical compositions were rich in caryophyllene120573-cadinene 120572-selinene and phytol [15 16] The insecticidenature of essential oils of P angolensis and P quadrifoliamanifested by contact adult insects S cerealellamay be linkedto the main volatile compounds extracted reportedly actingalone or in synergy with otherminor constituents In fact theessential oil of the leaves of P angolensis contains monoter-penes and oxygenated hydrocarbon such as 120572-pinene andOct-1-en-3-ol and Linalool which insecticidal properties havebeen demonstrated against Tribolium confusum Triboliumcastaneum Sitophilus zeamais Callosobruchus maculatusand Rhyzopertha dominica [11 17]

The repulsion rates evaluated of P angolensis essentialoil showed a greater repellent activity against adults ofAngoumois grain moth than P quadrifolia essential oilbelonging respectively to the repulsive V and IV classaccording to MC-Donald and Guyr [14] ranking This can beexplained by the strong offensive odor of both oil and confirmthe use of the leaves by farmers in protecting their grain insome parts of Benin Insect repellent effect of the extract ofP angolensis volatile proved more interesting than that of Pquadrifolia despite having a very low yield of essential oil

The use of essential oils in the preservation of cerealsis increasingly recognized [18] Essential oils are nowadaysknown as neurotoxins acutely interfering with octopamin-ergic transmitters in Arthropods [19] Different doses ofoils caused highly significant mortality (119875 lt 0001) incomparison with the mortality rates in the controls Paddyrice is the most common form of conservation only Scerealella species is likely to multiply in the paddy rice whichconfirms the results of previouswork [20] Increasing doses ofoil also reduced significantly the emergence of adults 50 daysafter infestation rates Such a reduction in emergence ratesresults from themanifestation of ovicidal or larvicidal volatileextracts which have destroyed the development of some eggs


Table 1 Yield and chemical composition of essential oil of P angolensis and P quadrifolia leaves

N∘ Name of the compound RI Premna angolensis Premna quadrifolia() ()

1 (Z)-3-Hexen-1-ol 850 10 mdash2 120572-Pinene 933 50 mdash3 Sabinene 965 mdash 374 120573-Pinene 970 mdash 065 Octen-3-ol 973 280 327 Octan-3-one 977 27 mdash8 Myrcene 987 03 mdash9 Octan-3-ol 991 14 mdash10 120572-Phellandrene 996 mdash 1011 Limonene 1022 mdash 1912 120573-Phellandrene 1023 mdash 0613 p-Cymene 1024 05 0814 Eucalyptol 1025 mdash t15 Limonene 1027 14 mdash16 (E)-120573-Ocimene 1039 mdash 0117 120574-Terpinene 1051 mdash 0118 (Z)-Sabinene hydrate 1064 mdash 0119 Linalool 1096 33 mdash20 Triacetonamine 1108 mdash 2821 Terpinen-4-ol 1174 25 0522 Naphthalene 1178 mdash 0923 120572-Terpineol 1188 mdash 0124 Methyl salicylate 1194 22 mdash25 Nerol 1221 t mdash26 Ethyl salicylate 1266 t mdash27 120572-Cubebene 1339 mdash 0228 120572-Ylangene 1362 mdash 0129 120572-Copaene 1369 mdash 1730 120573-Elemene 1374 14 2131 (E)-120573-Damascenone 1384 12 mdash32 120572-Gurjunene 1400 mdash 2033 120573-Caryophyllene 1414 mdash 13134 120574-Elemene 1421 mdash 1135 (E)-120573-Caryophyllene 1424 135 mdash36 (E)-120572-Bergamotene 1424 mdash 0837 Aromadendrene 1443 06 mdash38 (Z)-120573-Farnesene 1443 mdash 0239 Alloaromadendrene 1454 mdash 0440 120572-Humulene 1458 64 2941 Selina-411-diene 1466 mdash 1142 Germacrene-D 1475 mdash 8943 120573-Selinene 1486 06 1644 120574-Patchoulene 1490 28 mdash45 120572-Bulnesene 1490 mdash 1846 120572-Farnesene 1494 mdash 0847 120575-cadinene 1499 09 mdash48 120573-Bisabolene 1499 mdash 1949 Germacrene A 1501 mdash 02


Table 1 Continued


50 120574-Cadinene 1506 mdash 0951 (Z)-Calamenene 1512 mdash 0452 120575-Cadinene 1525 04 3353 Germacrene B 1553 mdash 1154 (E)-Nerolidol 1562 07 mdash55 Epiglobulol 1565 22 mdash56 Spathulenol 1571 mdash 0257 Caryophyllene oxide 1577 mdash 1358 Globulol 1590 112 mdash59 Humulene epoxide II 1616 09 mdash60 Epi-120572-muurolol 1649 mdash 1261 Selin-11-en-4-120572-ol 1653 mdash 1562 120572-Cadinol 1658 10 mdash63 Intermedeol 1673 03 mdash64 Phytol 2048 37 49

Total () 961 91Yield () 0056 0102

Hydrogenated monoterpenes () 86 89Oxygenated monoterpenes () 92 43Hydrogenated sesquiterpenes () 266 655Oxygenated sesquiterpenes () 20 91

Oxygenated aliphatic compounds () 317 32t (traces) = 01 RI retention index

Table 2 Rate of S cerealella death of emergence of number of rice attacked (119860) and of weight loss (119861) provoked by P angolensis essentialoil in contact method

Dose (120583LmLminus1) Mortality Emerged 119860 () 119861 ()0 1878 plusmn 007b 8571 plusmn 003a 2519 plusmn 241b 22726 plusmn 1955b

3 9012 plusmn 030a 107 plusmn 006b 037 plusmn 001c 0003 plusmn 0048c

5 9426 plusmn 017a 040 plusmn 011c 032 plusmn 0020c 0046 plusmn 0028c

10 100 plusmn 000a 033 plusmn 005cd 026 plusmn 002c 0003 plusmn 0024c

15 100 plusmn 000a 013 plusmn 010d 015 plusmn 002c 0000 plusmn 0020c

Probability lt0001lowastlowastlowast lt0001lowastlowastlowast lt0001lowastlowastlowast lt0001lowastlowastlowast

CV () 1375 1058 1876 16850 ethanol treatment (control) lowastlowastlowastvery highly significant difference (01)The averages followed by the same letter were not significantly different at the beginning of 5

Table 3 Rate of S cerealella death of emergence of number of rice attacked (119860) and of weight loss (119861) provoked by P quadrifolia essentialoil in contact method

Dose (120583LmLminus1) Mortality Emerged 119860 () 119861 ()0 1082 plusmn 017d 8962 plusmn 001a 2544 plusmn 093b 2431 plusmn 028b

3 5796 plusmn 005c 210 plusmn 004b 061 plusmn 005c 001 plusmn 000c

5 7038 plusmn 000bc 162 plusmn 008b 048 plusmn 005c 005 plusmn 002c

10 7675 plusmn 012ab 128 plusmn 005b 035 plusmn 004c 004 plusmn 003c





Table 4 Percentage () of the repellence with olfactometer of theessential oil of the leaves of P angolensis and P quadrifolia on adultSitotroga cerealella

Dose () Repellency rate () (plusmnSD)P angolensis P quadrifolia

01 6711 plusmn 1095 6555 plusmn 128605 88 plusmn 1095 8355 plusmn 9241 96 plusmn 894 9155 plusmn 1158Mean (plusmnSD) 8370 plusmn 116 8022 plusmn 1331Repellence class V IV

or larvae probably due to sesquiterpenoid compounds oroxygenated aliphatic and synergy with minor compounds

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

The authors are grateful to Polytechnic School of Abomey-Calavi (Benin) for financial support They are also thankfulto Professor Aristide ADOMOU from National Herbariumof Benin for plants identification The authors thank DrNoudogbessi that made the chemical analysis of essential oilsin the laboratory of Professor Chantal Menut in France (mayhe rest in peace)

References

[1] J W Kadareit ldquoFlowering plants-dicotyledonsrdquo inThe Familiesand Genera of Vascular Plants K Kubitzki Ed vol 7 p 449Springer 2004

[2] J Lovett and G P Clarke ldquoPremna schliebeniirdquo in IUCN 2006Red List of Threatened Species 1998

[3] A Hymavathi K S Babu V G M Naidu S R Krishna P VDiwan and J M Rao ldquoBioactivity-guided isolation of cytotoxicconstituents from stem-bark of Premna tomentosardquo Bioorganicand Medicinal Chemistry Letters vol 19 no 19 pp 5727ndash57312009

[4] H Sudo T Ide H Otsuka E Hirata A T TakushiShinzatoand Y Takeda ldquoMegastigmane benzyl and phenethyl alcoholglycosides and 4 41015840-dimethoxy-120573-truxinic acid catalpol diesterfrom the leaves of Premna subscandens MERRrdquo Chemical andPharmaceutical Bulletin vol 48 no 4 pp 542ndash546 2000

[5] J Desrivot J Waikedre P Cabalion et al ldquoAntiparasiticactivity of some new caledonian medicinal plantsrdquo Journal ofEthnopharmacology vol 112 no 1 pp 7ndash12 2007

[6] K P Devi M S Ram M Sreepriya G Ilavazhagan andT Devak ldquoImmunomodulatory effects of Premna tomen-tosa extract against Cr (VI) induced toxicity in spleniclymphocytesmdashan in vitro studyrdquo Biomedicine amp Pharmacother-apy vol 57 no 2 pp 105ndash108 2003

[7] British Pharmacopoeia 11 P A HMSO London UK 1980[8] R P Adams Identification of Essential Oil by Ion Trap Mass

Spectrometry Academy Press New-York NY USA 1989

[9] P Rosch J Popp and W Kiefer ldquoRaman and surface enhancedRaman spectroscopic investigation on Lamiaceae plantsrdquo Jour-nal of Molecular Structure vol 480-481 pp 121ndash124 1999

[10] A A Swigar and R M Silverstein Monoterpenes InfraredMass NMR Spectra and Kovats Indices Aldrich ChemMilwau-kee Wis USA 1981

[11] J P Noudogbessi M Keke F Avlessi D Kossou and D CK Sohounhloue ldquoEvaluation of the insecticidal larvicidal andovicidal effects on Callosobruchus maculatus of essential oilsof Cymbopogon giganteus and of Xylopia aethiopicardquo ScientificStudy amp Research vol 10 no 4 pp 337ndash338 2009

[12] CU Panthenius ldquoEtat des pertes dans les systemes de stockagesdumaıs au niveau des petits paysans dans la regionmaritime auTogordquo Fiche technique 83 GTZ 1988

[13] Centre drsquoEtude et drsquoExperimentation du Machinisme Agri-cole et Tropical (CEEMAT) Conservation des Grains enRegions Chaudes Techniques Rurales en Afrique Centre drsquoEtudeet drsquoExperimentation du Machinisme Agricole et Tropical(CEEMAT) Paris France 1988

[14] L MC-Donald and H D Guyr ldquoPreliminary evaluation ofnew candidate materials as toxicants repellents and attractantsagainst stored product insectsrdquo Marketing Research Report882 Agricultural Research Service United State Department ofAgriculture Washington DC USA 1970

[15] P K Renjana and J E Thoppil ldquoLarvicidal activities of the leafextracts and essential oil of Premna latifolia roxb (verbenaceae)against Aedes albopictus Skuse (Diptera Culicidae)rdquo Journal ofApplied Pharmaceutical Science vol 3 no 6 pp 101ndash105 2013

[16] C T Sadashiva P Sharanappa Y Naidoo and I BalachandranldquoChemical composition of essential oil from the leaves ofPremna coriacea Clarkerdquo African Journal of Biotechnology vol12 no 20 pp 2914ndash2916 2013

[17] G K Ketoh H K Koumaglo and I A Glitho ldquoInhibi-tion of Callosobruchus maculatus (F) (Coleoptera Bruchidae)development with essential oil extracted from Cymbopogonschoenanthus L Spreng (Poaceae) and the wasp Dinarmusbasalis Rondani (Hymenoptera Pteromalidae)rdquo Journal ofStored Products Research vol 41 pp 363ndash371 2006

[18] A Smith-Palmer J Stewart and L Fyfe ldquoThe potential applica-tion of plant essential oils as natural food preservatives in softcheeserdquo Food Microbiology vol 18 no 4 pp 463ndash470 2001

[19] N L Tatsadjieu P M J Dongmo M B Ngassoum F-X Etoaand C M F Mbofung ldquoInvestigations on the essential oil ofLippia rugosa from Cameroon for its potential use as antifungalagent againstAspergillus flavus Link ex Friesrdquo FoodControl vol20 no 2 pp 161ndash166 2009

[20] A Togola P A Seek I A Glitho et al ldquoEconomic losses frominsect pest infestation on rice stored on-farm in Beninrdquo Journalof Applied Sciences vol 13 no 2 pp 278ndash285 2013

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and repellent activities were tested on the Angoumois grainmoth Sitotroga cerealella (L) (Lepidoptera Gelechiidae) inrice storage

2 Material and Methods

21 PlantMaterial andDistillation of the Volatile ConstituentsLeaves of P angolensis and P quadrifolia were collectedfrom the municipality of Come by the authors and certifiedat the National Herbarium of Abomey-Calavi UniversityThe collected plant materials were stored in the laboratorybetween 18 and 20∘C in the shade of the sunlight throughoutthe period of extraction The essential oils were obtained byhydrodistillation of the leaves (450 g) for five hours usinga Clevenger-type extractor according to British Pharma-copoeia method [7] They were then dried over anhydroussodium sulfate and analyzed by GCMS

22 Test Organisms Individuals of S cerealella used formass rearing for this study come from the reserve WARDA(Benin) Rearing conditionswere119879= 29plusmn2∘C RH= 70plusmn10

23 Analysis of the Volatile Constituents

231 GCMS The essential oils were analysed on a Hewlett-Packard gas chromatograph Model 7890 coupled to aHewlett-Packard MS Model 5875 equipped with a DB5 MScolumn (30mtimes 025mm 025 120583m) programming from 50∘C(5min) to 300∘C at 5∘Cmin 5min hold Helium was used ascarrier gas (10mLminminus1) injection was made in split mode(1 30) injector and detector of temperature were at 250 and280∘C respectivelyTheMSworking in electron impactmodewas at 70 eV electron multiplier was 2500 eV ion sourcetemperature was 180∘C mass spectra data were acquired inthe scan mode inmz range 33ndash450

232 GCFID Theessential oils were analyzed on aHewlett-Packard gas chromatograph Model 6890 equipped with aDB5 MS column (30m times 025mm 025 120583m) programmingfrom 50∘C (5min) to 300∘C at 5∘Cmin 5min hold Hydro-gen was used as carrier gas (10mLminminus1) injection was insplit mode (1 60) injector and detector temperature were280 and 300∘C respectively The essential oil was diluted inhexane 130 The compounds assayed by GC in the differentessential oils were identified by comparing their retentionindices with those of reference compounds in the literatureand confirmed by GC-MS by comparison of their massspectra with those of reference substances [8ndash10]

24 Test All tests were performed at a temperature of 29plusmn2∘Cand natural photoperiod with relative humidity 70 plusmn 10

25 Contact Toxicity Tests with Essential Oils Bioassays wereperformed using the method described by Noudogbessi et al[11] The temperature of the test medium ranged from 25∘Cto 31∘C and relative humidity was 80 Five concentrations(0 3 5 10 and 15 120583LmLminus1) depending on the selected

75 cm

4 cm

Figure 1 Olfactometer set-up An individual female Angoumoisgrain moth was introduced in the center of the tube Her positionwas recorded after 15min since introduction Treated and controlpaddy rice were positioned at either end of the tube

chemical profile of each essential oil in absolute ethanolsolution were tested The plant material paddy rice grainsof the variety ldquoIR841rdquo was treated with each essential oilAfter 24 h 5 males and 10 females of S cerealella aged 0 to24 h were deposited on the treated plant material Overall5 doses times 3 repeats times 2 types of oil = 30 experimental units(containers) were implemented The control sample treatedonly with ethanol was also infested by S cerealella Adultmortality was monitored 96 h after exposure to the essentialoils Then the insects were separated from the grains Theemergence of new insects was then observed at intervals of24 h in the experimental units to the 50th day after infestationAssessment concerned the rate of infested grains and weightloss of grains of rice

To assess the S cerealella damage on the rice seeds aftertreatment the substrate grains were collected counted andweighed The rate of weight loss of dry material of the grainswas determined according to MCP (Method of CountingandWeighing) andMSVW (Method of Standard VolumetricWeight) [12] Two criteria of assessment of the damage arecommonly used the percentage of attack of grains (119860) andpercentage of weight loss (119861) [13] respectively calculatedby the equations

119860 = NaNa +Ns

times 100

119861 = PsNa minus PaNsPs (Na +Ns)

times 100

(1)

where Na is number of attacked grains Ns is number ofhealthy grains Pa is weight of damaged grains and Ps isweight of healthy grains

26 Repellency Tests The repellent action of the plants wastested in an olfactometer (Figure 1) consisting of a 75 cmglasstube of 4 cm in diameter with a 29 mm hole in the middleAt either end of the tube a small jar was placed containingeither 100 g containing paddy rice treated with only ethanolor 100 g of paddy rice seedsmixed with a solution of essentialoil at concentrations 01 05 and 1 The hole in themiddle was covered with gauze whereas the ends of the tubewere closed by putting a plastic petri dish against them Airwas gently (ca I mls) sucked away from the centre of the tubeto prevent the accumulation of plant odor in the tube

Ten female S cerealella aged 24 h to 48 h were releasedone after the other in themiddle of the tube through the holeInsect behavior was observed and the position was recordedfor 15min Both volatile extracts were tested once before







3 Results





4 Discussion









Table 1 Continued



Total () 961 91Yield () 0056 0102


























Acknowledgments


References




























Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology







3 Results





4 Discussion









Table 1 Continued



Total () 961 91Yield () 0056 0102


























Acknowledgments


References




























Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology






Table 1 Continued



Total () 961 91Yield () 0056 0102


























Acknowledgments


References




























Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


Table 1 Continued



Total () 961 91Yield () 0056 0102


























Acknowledgments


References




























Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology








Acknowledgments


References




























Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology








Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Documents

Premna cerealella - Hindawi Publishing Corporationdownloads.hindawi.com/archive/2015/319045.pdfResearch Article Chemical Composition and Insecticidal and Repellent Effect of Essential