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Preventive, curative and persistent activities of Lantana camara and Psidium guajava essential oils against Prostephanus truncatus (Horn).

Introduction

The protection and the conservation of foodstuffs after harvest remain a major concern in the African countries in the south of Sahara and in Benin in particular [1, 2]. In the optics to stimulate and to make profitable the maize production, one of the important foodstuffs for human consumption in Benin, many efforts have been made in the agricultural structures procurement in synthetic pesticides, in fertilizers and in the improvement of the seed varieties. But these efforts upstream and which are not supported swallows at the level post-harvest are generally destroyed by some predators that P. truncatus who commit their crimes from the fields and pursue them into the storage structures. The weight losses are estimated between 15 to 35% of the harvests after six to eight months of storage [3, 4, 5, 6]. P. truncatus, also called the Large Grain Borer is a typical pest of the maize [7]. It attacks the whole grains by drilling husk or by digging a main gallery through the grain to penetrate there. The eggs are put down by females in the perpendicular changing rooms to the main galleries, dug in the grain, in which develop larvae [8, 9]. In rural farms, the control methods used by the farmers against this pest are mainly based on endogenous practices and the use of chemical insecticides. Indeed, the farmers seem to be convinced that these pesticides too expensive and sometimes prohibited are efficient against the Larger Grain Borer [10]. The impact on humans and on environment owed to the misuse or by ignorance, added to the socio-economic difficulties generated by their sale, distribution and the storage in countryside and urban city is a thought- provoking approach. Hardly of this report, the recourse to biopesticide by the essential oils use seemed a cheaper alternative, credible, with little otherwise no major impact on the environment [11-13]. It is in this idea order that we approach this work which objective is to study the chemical compositions of L. camara and P. guajava essential oils and to evaluate their effects on P. truncatus.

Experimental

Plant material

The Lantana camara leaves in the fresh state were collected on the Abomeycalavi tray in June 2006 and those of P. guajava to Adjarra (Avrankou municipality) in May 2007. They were identified and certified to the Abomey- Calavi University National Herbarium and preserved in the laboratory between 18 and 20 [degrees] C in the shade of the sunlight during all the extractions period. The maize grains "Massahouekoun", local variety, were collected to Zoundji (Akpro Misserete municipality). They were kept in a cold room at 6[degrees]C throughout the experimental period.

Animal material

P. truncatus adults were taken to the International Institute of Tropical Agriculture (IITA / Benin). They were bred and maintained in the laboratory since January 2006 (T = 28[degrees]C [+ or -] 2[degrees]C, Relative Humidity = 80%). The insects selected for the realization of the biological assays were older than 60 days.

Chromatographical analysis

The essential oils were extracted by hydrodistillation (250 to 300g) for three hours using a Clevenger type apparatus. The volatile extracts were collected by simple decantation and dried over anhydrous sodium sulphate.

* GC/MS: The essential oils were analysed on a Hewlett-Packard gas chromatograph Model 7890, coupled to a Hewlett-Packard MS model 5875, equipped with a DB5 MS column (30m X 0, 25mm; 0, 25 [micro]m), programming from 50[degrees]C (5 min) to 300[degrees]C at 5[degrees]C/mn, 5 min hold. Helium as carrier gas (1, 0 mL/min); injection in split mode (1:30); injector and detector temperature, 250 and 280[degrees]C respectively. The MS working in electron impact mode at 70 eV; electron multiplier, 2500V; ion source temperature, 180[degrees]C; mass spectra data were acquired in the scan mode in m/z range 33-450.

* GC/FID: The essential oils were analysed on a Hewlett-Packard gas chromatograph Model 6890, equipped with a DB5 MS column (30m X 0, 25mm; 0, 25um), programming from 50[degrees]C (5min) to 300[degrees]C at 5[degrees]C/min, 5min hold. Hydrogen was used as carrier gas (1, 0 mL/min); injection in split mode (1:60); injector and detector temperature, 280 and 300[degrees]C respectively. The essential oil is diluted in hexane: 1/30.

The compounds assayed by GC in the different essential oils were identified by comparing their retention indices with those of reference compounds in the literature and confirmed by GC-MS by comparison of their mass spectra with those of reference substances [14-16].

Preventive effect

The maize was arranged under three forms of conservation (maize with husk, maize without husk, shelled maize) and placed in the glass bottle. Each form of conservation independently of the others was subject to each of the doses (0 mL, 0.1 mL, 0.15 mL, 0.2 mL) in three repetitions in a complete random block device. Thus, 36 experimental units were organized for all three forms of conservation. Forty-eight hours later, three insects adult (two females and one male) [11], of the same age (60 days) and taken from the mass rearing were deposited on the maize contained in each glass bottle. During the observation period, the females laid eggs. In every twenty-four hours, an insect which died was replaced and recorded. At the end of six days, the insects were removed and the maize grains so infested were put in observation during 40 days for the control of the emergence rhythm of the offspring. In the control treatment no taste of essential oil has been filed. The data registered during this control have focused on:

* the number of insects dead and recorded during six days

* the number of emergent insects per day for 40 days.

Curative effect

The maize was treated with the essential oil after six days of infestation. The measured data were identical to those of the preventive essay.

Persistent effect

The persistent effect was evaluated by infesting samples of maize treated with essential oil 15 days previously. In each glass bottler, three insects (two females and one male) were introduced and followed every 24 hours for 40 days. During this monitoring, an insect which died was recorded and was not any more replaced. After 40 days the insects still alive were entirely rid of the various forms of conservation. The data measured during this essay were:

* the number of insects dead at the end of 40 days

* the number of emergent insects at the end of 40 days.

Statistical analysis

The results from the observations were treated statistically by the method of analysis of variance (ANOVA) by means of the software SAS V 9.1 [17]. The recorded data underwent a transformation square root in order to stabilize the variances and to standardize the population. The formula used is: [square root of ((X + 0, 5))] with X = given rough (emerged or died) [18]. Then, it was proceeded a structuring of the averages using the test of Newman and Keuls [19]. The results of the statistical tests are regarded as significantly different, when the probability of ANOVA is lower or equal to 5%.

Results and discussion

Yields and chemical compositions of L. camara and P. guajava leaves essential oils:

The contents of L. camara and P. guajava leaves essential oils were respectively 0.023% and 0.54%. According to these two values, P. guajava leaves, at the harvest time, contained more essential oil than those of L. camara. The chemical compositions of the two plants were given in the following table (Table I). The chromatographic analyzes performed showed 37 compounds representing 95.6% of the total weight of P. guajava essential oil whereas in Lantana camara volatile extract, 41 compounds corresponding to 96.8% of the weight of this essential oil were identified. L. camara essential oil was richer in hydrogenated terpenes (77.1%) than that of P. guajava (56.0%). The rate in oxygenated compounds of the P. guajava essential oil was 37.0%. [beta]-caryophyllene (19.3%), sabinene (15.9%), (Z)-[beta]-ocimene (11.2%), [alpha]-humulene (6.5%), germacrene-A (5.8%) were the major compounds of L. camara essential oil. These same compounds, especially [beta]-caryophyllene, sabinene and [alpha]-humulene had been previously reported in L. camara leaves essential oils studied by Rabendra Balandra in 2011 [20], Oluwadayo Sonibare and Effiong in 2008 [21]. On the other hand the present chemical composition differs from that studied by Sousa et al. in Brazil in 2012 which mentions, except the [beta]-caryophyllene, other major compounds namely bicyclogermacrene (26.1%), germacrene-D (19.2%), valencene (12.0%) [22]. This difference was more significant in comparison for the major compounds ([beta]-caryophyllene: 13.57%, [alpha]-caryophyllene: 11.76%, 10.88% germacrene-D, isocaryophillene: 9.59% [gamma]-muurolene: 6.85% [gamma]-elemene: 5.65%) of Bangladesh L. camara leaves essential oil [23]. P. guajava essential oil contained epi-[beta]-bisabolol (11.7%), [beta]-caryophyllene (11.0%), [beta]-bisabolene (7.5%), [beta]-curcumene (4.2 %) as dominant compounds. The major compounds reported in the present work were significantly different from those reported by Fasola et al. in the P. guajava stem-bark volatile extract of Nigeria. Indeed, the main compounds identified in this extract were hydrocarbons, amines, amides, esters accompanied by 3, 6- dioxa-2, 4, 5, 7-tetraoctane, 2, 2, 4, 4, 5, 5, 7, 7-octamethyl (11.67%) and cyclononane (10.66%) [24]. It is the same of the volatile extract of the leaves of P. guajava investigated in India and of chemical composition dominated by methyl hexadecanoate (9.32%), propyl benzene (9.52%), methyl tetradecyl acetate (12.86%), 3, 6, 9-nonadecatriene (15.37%), methyl octadecanoate (22.18%), methyl 2, 6, 10-trimethyltridecanoate (28.86%) [25].

Comparative effects of L. camara and P. guajava essential oils on P. truncatus living on maize:

The tables II to VII below showed the statistical averages induced by each essential oil on P. truncatus alive on a shape of preservation given of the maize. In these tables, the statistical averages were compared by column. The Table II shows the evolution of the mortality rates evolution and emergence engendered by P. truncatus in the presence of L. camara essential oil. The essential oil of L. camara, did not exhibit a toxic effect on the P. triuncatus adults existence during the oviposition period (six days) for the three conservation methods evaluated (Table II). The mortality averages recorded for the doses 0.1; 0.15 and 0.2 mL was significantly similar to those of the control. At the emergence, the applied doses (0.1; 0.15 and 0.2 mL) caused low rates of emergent insects, but different from the control. However, L. camara essential oil has prevented the development of eggs put down by the females of P. truncatus and of their larvae newly formed. This essential oil thus possessed ovicidal and larvicidal effects. The local variety of maize without husk "Massahouekoun" submitted to the P. truncatus pressure by different methods was treated with L. camara essential oil. The low averages of mortality recorded and aggregated in this Table III showed a relatively low toxicity of L. camara essential oil on P. truncatus adult. The L. camara volatile extract thus showed a weak insecticidal effect against P. truncatus adult living on maize without husk. This finding was in adequacy with the chemical composition (Table I) of L. camara essential oil poor in phenolic compounds often responsible for insecticidal and repellent activities as it is the case for the P. racemosa and C. odorata essential oils in a previous study [26]. The new insects rates recorded at the end of 40 days were low contrary to the control according to the mean values obtained in curative, preventive and persistent methods. The delay in the emergences development would be the result of the molecules with ovicidal and / or larvicidal action, acting in synergy or not, present in L. camara essential oil. In the table IV where were recorded the mean values of P. truncatus mortality and emergence, the insecticidal effect performance of L. camara essential oil was rather remarkable. In the curative treatment, the mortality averages were not significantly different. It's the same in preventive and persistent methods. On the other hand, the treatments 0.1, 0.15, 0.2 mL have produced so much of emerging for every conservation method. In curative method, the lowest emergence (0.33) was produced by the dose of essential oil (0.2 mL) that is the sufficient quantity to stop the infestation of the maize grains after the oviposition. For the doses 0.1 mL and 0.2 mL in a preventive mode, the averages emergence 13.33 and 14.33; statistically similar, were very different from that obtained (1.67) to the dose 0.15 mL. This difference between the averages was probably due to an egg deposited by P. truncatus females during the treatment 0.15 mL. In the persistent method, the emergent rates were low and were the consequence of the time effect (15 days) of L. camara essential oil on maize grains which offered no more rather good conditions for the development of eggs produced. The insect's mortality during the oviposition period was less important at the level of the substrates treated with P. guajava essential oil in curative, preventive and persistent methods (Table V). The registered averages do not exhibit the volatile extract insecticidal or repellent effect on adult P. truncatus. In curative treatment, the rate (39.00) of emergent generated by the processing 0.1 mL was significantly different from the control (143.33) and very high compared to the averages produced by the doses 0.15 and 0.2 mL. However, the emergences engendered in the maize ears treated with these last doses (0.15 and 0.2 mL) were weak and statistically identical after 40 days. An ovicidal and / or larvicidal of P. guajava essential oil was thus to be indicated of this fact. In the analysis of the preventive method results, the emergence averages significantly different from that of the control were observed for the quantities 0.1, 0.15 and 0.2 mL. P. guajava essential oil showed for these last treatments a low ovicidal and larvicidal against P. truncatus eggs and larvae. The same observations was made in persistent method. The results in the table VI concerned the mortalities and the emergence averages of P. truncatus living on maize without husk. In curative method, enough new insects were recorded although the doses 0.15 and 0.2 mL were important. This observation was the consequence of the development of certain eggs produced by some insects inside the raids. These raids were also a physical factor facilitating the increase of the emergent insects in preventive and persistent methods. The statistical results of the table VII shows very low averages of adult mortality of P. truncatus living on the shelled maize for all the forms of conservation. The absence of raid and husk to which was added the low anti-infective character sesquiterpenes alcohol from P. guajava essential oil has contributed to the insect's survival during the oviposition period. The P. guajava essential oil showed neither insecticidal effect, nor repellent on P. truncatus. The females thus left eggs which produced later emergences according to the preservation mode used. In curative treatment, the statistical similarities noted between the average values for the doses 0.1, 0.15 and 0.2 mL were different from that of the control (P < 0.0004). The P. guajava volatile extract has demonstrated actions delaying the pest development. The trend seems to be the same for the persistent method but the effect of the doses 0.1, 0.15 and 0.2 mL has significantly reduced the registered averages. In the preventive method, the treatments 0.1 and 0.2 mL produced emergences statistically identical but different from those of the control and from the dose 0.15 mL. This result did not guarantee in our opinion an efficiency of the P. guajava essential oil in front of P. truncatus because the substratum shape was not enough adapted to the pest biology.

Conclusion

The control of maize pests, namely P. truncatus, was confronted, in recent years, for a resistance to synthetic insecticides (non-biodegradable and harmful to humans) of the devastating. Considering the prohibitive cost of these insecticides, their toxicity and their degrading action on the environment, the current trend aimed at the use of the phyto-bioactive extracts, little toxic, biodegradable, less expensive and proven effectiveness. It was about the essential oils of certain aromatic plants containing highly active molecules and having insecticidal, larvicidal and ovicidal properties. In the case of the current work, the chemical composition study of the essential oils showed, mainly, the presence of [beta]-caryophyllene in the L. camara leaves and epi-[beta]- bisabolol in those of P. guajava. The emergence averages high enough at the level of control substratum were probably due to the strong fertility of P. truncatus, the good fertility of its eggs and larvae efficient penetration in grains.maize. L. camara and P. guajava essential oils very rich in hydrogenated terpenes accompanied by low rates of oxygenated compounds affected the P. truncatus eggs hatching and the development of its newly formed larvae. These volatile extracts have shown ovicidal and larvicidal properties in all the forms of maize conservation.

References

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[8] Belle R. J. and Watters F. L., 1982, "Environmental factors influencing the development and rate of increase of Prostephanus truncatus on stored maize," J. Stored Prod. Res., 18, pp. 131-142.

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[10] Mailafiya D. M., Ayertey J. N., Cudjoe A. R., 2008, "Damage and weight loss potential of Prostephanus truncatus (Horn) Coleoptera: Bostrichidae) on sorghum grain: implication to cereal grain storage in sub-Saharan Africa," International Journal of Pure and Applied Sciences, 2, pp. 28-35.

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[12] Kossou K. D., 1992, "Sensibilite des bois de construction des greniers traditionnels aux attaques de Prostephanus truncatus (Horn) (Coleoptera: Bostrichidae)," Insect Sci. Applic., 13, pp. 435-439.

[13] Gakuru S. and Foua-bi K., 1995, "Effet compare des huiles essentielles de quatre especes vegetales contre la bruche du niebe (Callosobruchus maculatus Fab) et le Charancon du riz (Sitophilus oryzae L.)," Tropicultura, 13, pp. 143-146.

[14] Rosch P., Popp J., Kiefer W., 1999, "Raman and SERS Investigations on Lamiaceae," J. Mol. Struct., 121, pp. 480-481.

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[16] Swigar, A. A. and Silverstein, R. M., 1981, "Monoterpenes: infrared, mass, 1H-NMR, 13C-NMR spectra and kovats indices," Aldrich Chemical Company Inc., Wisconsin.

[17] Dagnelie P., 1975, "Theorie et Methodes statistiques. Applications agronomiques. Les Presses Agronomiques de Gembloux A.S.B.L.," Avenue de la faculte, 22-5800 Gembloux (Belgique), 2, pp. 109-152.

[18] Dagnelie P., 1975, "Theorie et Methodes statistiques. Applications agronomiques. Les Presses Agronomiques de Gembloux A.S.B.L.," Avenue de la faculte, 22-5800 Gembloux (Belgique), 2, pp. 362-363.

[19] Dagnelie P., 1975, "Theorie et Methodes statistiques. Applications agronomiques. Les Presses Agronomiques de Gembloux A.S.B.L.," Avenue de la faculte, 22-5800 Gembloux (Belgique), 2, pp. 245-250.

[20] Rabindra K. S., Balendra T., 2011, "Composition of Lantana camara leaf essential oil," International Journal of Pharmaceutical Research and Development, 3(7), pp. 51-55.

[21] Oluwadayo Sonibare O. and Effiong I., 2008, "Antibacterial activity and cytotoxicity of essential oil of Lantana Camara L. leaves from Nigeria," African Journal of Biotechnology, 7 (15), pp. 2618-2620.

[22] Sousa E. O., Almeida T. S., Menezes I. R. A., Fabiola F. G. R., Campos A. R., Lima S. G. and da Costa J. G. M., 2012, "Chemical Composition of Essential Oil of Lantana camara L. (Verbenaceae) and Synergistic Effect of the Aminoglycosides Gentamicin and Amikacin," Rec. Nat. Prod., 6(2) pp. 144-150.

[23] Chowdhury J. U., Nandi N. C. and Nazrul Islam Bhuiyan M. D., 2007, "Chemical Composition Of Leaf Essential Oil Of Lantana Camara L. From Bangladesh," Bangladesh J. Bot., 36(2), pp. 193-194.

[24] Fasola T. R., Oloyede G. K. and Aponjolosun B. S., 2011, "Chemical Composition, Toxicity And Antioxidant Activities Of Essential Oils Of Stem-Bark Of Nigerian Species Of Guava (Psidium Guajava Linn.)," EXCLI Journal, 10, pp. 34-43.

[25] Nisha K., Darshana M., Madhu G. and K Bhupendra M., 2011, "GC-MS Analysis and anti-microbial activity of Psidium Guajava (leaves) grown in Malva region of India," Int. J. Drug Dev. & Res., 3(4), pp. 237-245.

[26] Noudogbessi P. P., Kossou D., Sohounhloue D. C. K., 2008, "Composition Chimique et Proprietes Physico-Chimiques des Huiles Essentielles de Pimenta racemosa (Miller) et de Chromolaena odorata (L. Robinson) Acclimatees au Benin," J. Soc. Ouest-Afr. Chim., 026, pp. 11-19.

Jean-Pierre A. Noudogbessi (1), Elisabeth T. Zannou-Boukari (6), Leonce F. Dovonon (1), Alassane Youssao (1), Magloire Gbaguidi (1), Gilles Figueredo (2), Pierre Chalard (3, 4), Jean C. Chalchat (5), Dansou Kossou (6) and Dominique C. K. Sohounhloue (1) *

(1) Ecole Polytechnique d'Abomey-Calavi, Laboratoire d'Etude et de Recherche en Chimie Appliquee, Cotonou, 01. BP 2009, Benin

(2) Laboratoire d'Analyse des Extraits Vegetaux et des Aromes (LEXVA Analytique), 460 Rue du Montant, 63110 Beaumont, France

(3) Institut de Chimie de Clermont-Ferrand (ICCF), Ecole Nationale Superieure de Chimie de Clermont-Ferrand (ENSCCF), BP10448, F-63000 Clermont-Ferrand (France)

(4) UMR 6296, ICCF--CNRS, F-63171 Aubiere

(5) Universite Blaise-Pascal, Laboratoire de Chimie des Huiles Essentielles, Clermont-Ferrand--Campus des Cezeaux, 63177 Aubiere cedex, France

(6) Universite d'Abomey-Calavi / Faculte des Sciences Agronomiques / Departement de Production Vegetale, Cotonou, 01. BP 526, Benin

* ksohoun@bj.refer.org
Table I : Chemical compositions of the essential oils of P. guajava
and L. camara leaves

Constituent                          KI      Pg     Lc
                                                (%)

tricyclene                           929     1.7    0.4
[alpha]-pinene                       935     -      2.3
Camphene                             946     -      1.2
Benzaldehyde                         959     2.5    -
Sabinene                             973     -      15.9
[beta]-pinene                        979     -      1.9
6-methyl-5-hept-5-en-2-one           981     0.2    -
Myrcene                              991     0.2    1.8
p-mentha- 1(7),8-diene               999     0.3    2.5
[alpha]-terpinene                    1015    -      0.3
p-cymene                             1021    0.3    -
Limonene                             1028    20.7   0.2
1,8-cineole                          1029    0.1    1.6
(Z)-[beta]-ocimene                   1037    -      11.2
(E)-[beta]-ocimene                   1050    -      1.2
[gamma]-terpinene                    1057    -      0.6
cis-sabinene hydrate                 1070    -      0.9
Terpinolene                          1083    -      0.4
trans- sabinene hydrate              1098    -      0.9
cis-p-menth-2-en-1-ol                1122    -      -
Camphor                              1146    0.7    1.4
Borneol                              1171    -      0.2
terpinen-4-ol                        1177    -      1.9
[alpha]-terpineol                    1192    -      1.1
methyl geranate                      1316    0.2    -
Eugenol                              1348    -      1.3
[alpha]-copaene                      1374    3.9    0.4
[beta]-elemene                       1384    -      0.6
[beta]-caryophyllene                 1419    6.3    19.3
[beta]-copaene                       1428    -      0.4
[alpha]-trans-bergamotene            1429    0.3    -
[alpha]-humulene                     1454    1.2    6.5
Sesquisabinene                       1459    -      0.2
[gamma]-muurolene                    1472    0.5    -
germacrene-D                         1480    -      0.2
[beta]-selinene                      1488    3.1    -
[alpha]-selinene                     1491    2.8    -
bicyclogermacrene                    1493    -      1.4
germacrene-A                         1498    -      6.0
[delta]-cadinene                     1522    0.8    0.6
[delta]-cuprenene                    1546    -      0.9
germacrene-B                         1555    -      1.0
(E)-nerolidol                        1556    0.9    3.5
caryophyllene alcohol                1574    0.4    0.6
sesquisabinene (E)-hydrate           1578    -      0.6
caryophyllene oxide                  1580    2.7    0.8
P-copaen-4-[alpha]-ol                1584    0.3    -
guaiol                               1591    0.4    -
epi-globulol                         1606    0.4    -
humulene epoxyde II                  1607    1.2    0.8
1,10-di-epi-cubenol                  1619    0.3    -
1-epi-cubenol                        1631    3.7    2.1
[beta]-acorenol                      1635    2.4    -
epi-[alpha]-cadinol                  1639    -      0.4
epoxy-allo-alloaromadendrene         1633    1.6    -
epi-[alpha]-muurolol                 1641    1.4    -
[alpha]-muurolol                     1644    2.3    -
selin-11-en-4-[alpha]-ol             1660    9.9    -
14-hydroxy-9-epi-(E)-caryophyllene   1668    0.9    -
nerolidyl acetate                    1678    -      1.3
[alpha]-bisabolol                    1683    0.2    -
(2Z, 6Z)-farnesol                    1709    4.3    -
(2Z, 6E)-farnesol                    1710    10.0   -
(2E, 6E)-farnesol                    1742    6.0    -
benzyl benzoate                      1764    0.5    -
Total                                        95.6   96.8

Pg = Psidium guajava; Lc = Lantana camara; KI = Kovats Indice; exp =
experimental

Table II: averages of P. truncatus dead and emerged on the maize in
husk treated with L. camara essential oil

Essential oil                     curative method
 doses (mL)              died                       emerged

     0.0           0[+ or -]0(0.71)a      112.33[+ or -]10.52(10.60)a
     0.1        0.33[+ or -]0.33(0.88)a     0.33[+ or -]0.33(0.88)b
    0.15           0[+ or -]0(0.71)a        4.33[+ or -]3.38(1.89)b
     0.2           0[+ or -]0(0.71)a        1.33[+ or -]1.33(1.18)b
 Probability            0.44ns                   < 0.0001 ***
   CV (%)                19.92                       25.18

Essential oil                    preventive method
 doses (mL)              died                       emerged

     0.0        0.33[+ or -]0.33(0.88)a   111.00[+ or -]5.86(10.55)a
     0.1           0[+ or -]0(0.71)a        1.67[+ or -]0.88(1.39)b
    0.15           0[+ or -]0(0.71)a        0.67[+ or -]0.33(1.05)b
     0.2           0[+ or -]0(0.71)a        0.33[+ or -]0.33(0.88)b
 Probability            0.44ns                   < 0.0001 ***
   CV (%)                19.92                       12.68

Essential oil                    persistent method
 doses (mL)              died                       emerged

     0.0        2.00[+ or -]1.00(1.48)a   103.00[+ or -]1.73(10.17)a
     0.1        2.33[+ or -]0.33(1.68)a        0[+ or -]0(0.71)b
    0.15        2.67[+ or -]0.33(1.77)a     8.67[+ or -]6.76(2.52)b
     0.2        3.00[+ or -]0.00(1.87)a     0.33[+ or -]0.33(0.88)b
 Probability           0.6054ns                  < 0.0001 ***
   CV (%)                20.93                       29.05

ns = not significant at 5%; *** = very highly significant difference
(0.1%). The averages enter brackets arise raw data. The averages
followed by the same letter were not significantly different at the
beginning of 5% (Newman and Keuls test). CV = covariance

Table III: averages of P. truncatus dead and emerged on the maize
without husk treated with L. camara essential oil

Essential oil                    curative method
 doses (mL)              died                       emerged

     0.0           0[+ or -]0(0.71)a      112.00[+ or -]21.55(10.50)a
     0.1        0.33[+ or -]0.33(0.88)a    21.33[+ or -]9.06(4.47)b
    0.15        0.33[+ or -]0.33(0.88)a     7.33[+ or -]6.84(2.18)b
     0.2           0[+ or -]0(0.71)a       23.00[+ or -]13.57(4.11)b
 Probability            0.60ns                     < 0.01 **
CV (%) 26.64                                         42.55

Essential oil                    preventive method
 doses (mL)              died                       emerged

     0.0           0[+ or -]0(0.71)a       97.67[+ or -]4.84(9.90) a
     0.1           0[+ or -]0(0.71)a       19.67[+ or -]2.18(4.48)ab
    0.15        1.00[+ or -]1.00(1.09)a   23.00[+ or -]15.69(4.03)ab
     0.2           0[+ or -]0(0.71)a      19.67[+ or -] 19.17 (3.19)b
 Probability            0.44ns                      0.045 *
CV (%) 26.64             41.78                       47.41

Essential oil                    persistente method
 doses (mL)              died                       emerged

     0.0        1.33[+ or -]0.67(1.29)a   113.33[+ or -]13.84(10.63)a
     0.1        0.67[+ or -]0.67(0.99)a     4.00[+ or -]4.00(1.65)b
    0.15        1.67[+ or -]0.67(1.44)a    14.67[+ or -]7.42 (3.39)b
     0.2        2.00[+ or -]0.58 (1.56)a  26.33[+ or -]13.42 (4.44)b
 Probability            0.468ns                  < 0.0057 ***
CV (%) 26.64             32.83                       44.38

ns = not significant at 5%; *** = very highly significant difference
(0.1%); ** = very highly significant difference (1%); * = very
highly significant difference (5%). The averages enter brackets arise
raw data. The averages followed by the same letter were not
significantly different at the beginning of 5% (Newman and Keuls
test. CV = covariance

Table IV : averages of P. truncatus dead and emerged on the shelled
maize treated with L. camara essential oil

Essential oil                   curative method
 doses (mL)              died                      emerged

     0.0           0[+ or -]0(0.71)a      104.33[+ or -]4.81(10.23)a
     0.1        0.33[+ or -]0.33(0.88)a    10.00[+ or -]5.29(2.85)b
    0.15           0[+ or -]0(0.71)a       2.66[+ or -]1.45(1.64)b
     0.2           0[+ or -]0(0.71)a       0.33[+ or -]0.33(0.88)b
 Probability            0.44ns                   < 0.0001 ***
   CV (%)                19.92                      27.35

Essential oil                  preventive method
 doses (mL)              died                      emerged

     0.0           0[+ or -]0(0.71)a      114.67[+ or -]4.33(10.73)e
     0.1        1.33[+ or -]0.88(1.27)a    13.33[+ or -]7.79(3.21)b
    0.15        1.00[+ or -]0.58(1.17)a    1.67[+ or -]0.88(1.38)b
     0.2        0.33[+ or -]0.33(0.88)a   14.33[+ or -]14.33(2.67)b
 Probability            0.342ns                   0.0023 **
   CV (%)                39.19                      46.31

Essential oil                  persistent method
 doses (mL)              died                      emerged

     0.0        2.33[+ or -]0.67(1.65)a   87.33[+ or -]8.67(9.35)a
     0.1        1.67[+ or -]0.67(1.44)a    0.67[+ or -]0.67(0.99)b
    0.15        2.67[+ or -]0.33(1.77)a       0[+ or -]0(0.71)b
     0.2        1.33[+ or -]0.88(1.27)a    1.00[+ or -]0.58(1.17)b
 Probability           0.4642ns                  < 0.0001 ***
   CV (%)                26.19                      17.14

ns = not significant at 5%; *** = very highly significant difference
(0.1%); ** = very highly significant difference (1%). The averages
enter brackets arise raw data. The averages followed by the same
letter were not significantly different at the beginning of 5%
(Newman and Keuls test. CV = covariance

Table V : averages of P. truncatus dead and emerged on the maize in
husk treated with P. guajava essential oil

Essential oil                      curative method
 doses (mL)              died                       emerged

     0.0        0.33[+ or -]0.33(0.88)a   143.33[+ or -]4.48(11.99)a
     0.1        0.33[+ or -]0.33(0.88)a   39.00[+ or -]19.21(5.50)b
    0.15           0[+ or -]0(0.71)a       0.67[+ or -]0.33(1.05)c
     0.2        0.33[+ or -]0.33(0.88)a    0.33[+ or -]0.33(0.88)c
 Probability            0.802ns                   < 0.0003***
   CV (%)                30.94                       38.73

Essential oil                     preventive method
 doses (mL)              died                       emerged

     0.0        0.67[+ or -]0.67(0.99),   103.00[+ or -]14.42(10.12)a
     0.1        1.00[+ or -]0.00(L22)<    12.67[+ or -]12.67(2.54)b
    0.15        1. 67[+ or -]0.33(L46)<     6.00[+ or -]3.46(2.26)b
     0.2         2.00[+ or -]L15(L47)<     1.00[+ or -]1.00(1.09)b
 Probability            0.552ns                    0.0014 **
   CV (%)                34.83                       47.05

Essential oil                     persistente method
 doses (mL)              died                       emerged

     0.0        2.33[+ or -]0.67(1.65)a   122.00[+ or -]6.11(11.06)a
     0.1        2.00[+ or -]1.00(1.48)a   19.33[+ or -]19.33(3.02)b
    0.15        1.00[+ or -]1.00(1.09)a    7.67[+ or -]5.69(2.50)b
     0.2        3.00[+ or -]0.00(1.87)a    6.67[+ or -]6.67(1.98)b
 Probability           0.3571ns                   < 0.0056 **
   CV (%)                33.44                       52.77

ns = not significant at 5%; *** = very highly significant difference
(0.1%); ** = very highly significant difference (1%). The averages
enter brackets arise raw data. The averages followed by the same
letter were not significantly different at the beginning of 5%
(Newman and Keuls test. CV = covariance

Table VI : averages of P. truncatus death and emerged on the maize
without husk treated with P. guajava essential oil

Essential oil                      curative method
 doses (mL)              died                      emerged

     0.0        0.67[+ or -]0.67(0.99)a   98.67[+ or -]10.33(9.93)a
     0.1           0[+ or -]0(0.71)a       6.00[+ or -]3.46(2.26)b
    0.15        0.33[+ or -]0.33(0.88)a   23.67[+ or -]13.57(4.18)b
     0.2        0.33[+ or -]0.33(0.88)a   31.00[+ or -]15.87(4.93)b
 Probability            0.758ns                   < 0.025 *
   CV (%)                37.99                      45.65

Essential oil                     preventive method
 doses (mL)              died                      emerged

     0.0           0[+ or -]0(0.71)a      130.33[+ or -]17.33(11.39)a
     0.1        0.33[+ or -]0.33(0.88)a    13.33[+ or -]4.70(3.57)b
    0.15        1.33[+ or -]0.88(1.27)a    3.00[+ or -]2.08(1.67)b
     0.2        2.67[+ or -]1.76(1.61)a    9.00[+ or -]6.24(2.63)b
 Probability           0.2773ns                   0.0001 ***
   CV (%)                50.63                      29.78

Essential oil                     persistent method
 doses (mL)              died                      emerged

     0.0        0.67[+ or -]0.67(0.99)a   100.00[+ or -]11.93(9.99)a
     0.1        1.00[+ or -]0.58(1.17)a    5.33[+ or -]3.38(2.21)b
    0.15        0.33[+ or -]0.33(0.88)a    12.33[+ or -]6.49(3.13)b
     0.2        1.67[+ or -]0.88(1.38)a    4.67[+ or -]2.40(2.06)b
 Probability           0.6080ns                  < 0.0004 ***
   CV (%)                42.87                      33.51

ns = not significant at 5%; *** = very highly significant difference
(0.1%); * = very highly significant difference (5%). The averages
enter brackets arise raw data. The averages followed by the same
letter were not significantly different at the beginning of 5%
(Newman and Keuls test. CV = covariance

Table VII : averages of P. truncatus death and emerged on the shelled
maize treated with P. guajava essential oil

Essential oil                      curative method
 doses (mL)              died                       emerged

     0.0           0[+ or -]0(0.71)a      113.67[+ or -]9.74(10.67)a
     0.1           0[+ or -]0(0.71)a        6.33[+ or -]4.84(0.29)b
    0.15        0.33[+ or -]0.33(0.88)a    14.67[+ or -]6.89(3.53)b
     0.2           0[+ or -]0(0.71)a        5.33[+ or -]3.93(2.08)b
 Probability            0.44ns                   < 0.0004 ***
   CV (%)                19.92                       32.74

Essential oil                     preventive method
 doses (mL)              died                       emerged

     0.0        0.67[+ or -]0.67(0.99)a    97.67[+ or -]5.04(9.90)a
     0.1           0[+ or -]0(0.71)a        4.00[+ or -]2.08(2.00)c
    0.15        1.33[+ or -]0.67(1.29)a    11.67[+ or -]2.33(3.46)b
     0.2        1.00[+ or -]0.00(1.22)a     2.67[+ or -]2.18(1.56)c
 Probability           0.2568ns                   0.0001 ***
   CV (%)                33.82                       18.21

Essential oil                     persistente method
 doses (mL)              died                       emerged

     0.0           0[+ or -]0(0.71)a      104.67[+ or -]17.89(10.17)a
     0.1        0.67[+ or -]0.67(0.99)a     0.67[+ or -]0.67(0.99)b
    0.15        0.67[+ or -]0.33(1.05)a     3.00[+ or -]2.52(1.62)b
     0.2        2.00[+ or -]1.00(1.48)a     2.00[+ or -]1.23(1.43)b
 Probability           0.2764ns                  < 0.0001 ***
   CV (%)                42.06                       30.92

ns = not significant at 5%; *** = very highly significant difference
(0.1%). The averages enter brackets arise raw data. The averages
followed by the same letter were not significantly different at the
beginning of 5% (Newman and Keuls test. CV = covariance
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Author:Noudogbessi, Jean-Pierre A.; Zannou-Boukari, Elisabeth T.; Dovonon, Leonce F.; Youssao, Alassane; Gb
Publication:International Journal of Applied Chemistry
Article Type:Report
Geographic Code:6BENI
Date:Jan 1, 2013
Words:6170
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