Comparative Study Between ATOMES Novels, NOVOSECT SC21, ATO BED BUG and NEO-BOOST as a Bio-organic Solution in Managing Tomato Open Field Plantation in Hrajel Area in Lebanon

The increasing use of chemical insecticides has adversely effected the environment and increased insect resistance. Biopesticides have been noticed the potential to be an excellent alternative to chemicals to reduce the negative impacts to human health and the environment. Tomato (Lycopersicon esculentum) is the second most important vegetable crop worldwide due to its nutritional importance. The effect of NOVOSECT SC21 (0.5 L/200 L), ATO BED BUGS (1 L/200 L) and NEO-BOOST (1 kg/200 L) against Tuta absoluta, Liriomyza trifolii and Alternaria solani was studied. A complete randomized block design (CRBD) was used with three replications, three treatments and one control in Hrajel area in Lebanon in the summer of 2020. We evaluated the level of infestation and larval mortality level of Tuta absoluta, Liriomyza trifolii and Alternaria solani infection 24, 48 and 72 hr after the application. Results showed that NOVOSECT SC21 (Mix of metabolites of the Bacillus F.D. 777) was the most efficient in decreasing the infestation and inducing larval mortality level of T. absoluta and L. trifolii and the fungal infection induced by A. solani with significant difference with time, followed respectively by ATO BED BUGS and finally NEO-BOOST.


Introduction
Vegetables are one of the most important crops in agriculture and tomato (Lycopersicon esculentum) is considered as one of the most cultivated and consumed vegetable crops in Lebanon, with a production of 300,157 tons in 2018 and a cultivated area of 3700 ha according to IDAL (Investment Development Authority in Lebanon) (IDAL, 2020). Over the years, the increased demand for vegetables has resulted in the increase in land cultivation and the adoption of intensive farming in greenhouses. However, the increasing pressure on the soil and the excessive use of chemical fertilizers weakens the soil and disturbs its microbiological balance leading with time to weaken seedlings, which makes them susceptible to diseases and pests.
The intensive use of chemical insecticides have serious drawbacks, including reduced profits from high insecticide costs, destruction of insect pest's natural enemy populations, the build up of insecticide residues on tomato fruit. In addition, the high use of chemical insecticides can adversely affect the environment and increase the insecticide resistance to many insects, including Tuta absoluta and Liriomyza trifolii, which are among the most important tomato pests in Mount Lebanon, causing severe problems to tomato crops and significant losses in production that could reach 100% if not controlled (Devine et al., 2007;Giorgini et al., 2018;MOA, 2016). The need to adopt new technologies and good agricultural practices should be taken into consideration in order to lower the production costs, lower the insecticide residues in the product, protecting the environment, and obtaining effective insect control (IDAL, 2020). Natural products are an excellent alternative to synthetic pesticides as they help reducing their negative impacts on human health, while protecting the environment and jas.ccsenet. beneficial the enviro include su pesticide, have gener purpose of F.D. 777), and morta field in Hr

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Effect of Biopesticides on Tuta absoluta Infestation
The results obtained using T-test between 24 and 48 hr, indicate a reduction in Tuta absoluta level of infestation/plant: the mean after 24 hours is 1.36±0.49, while the mean at 48 hr is 0.91±0.22. The paired samples correlation indicates that there is a moderate correlation between 24 and 48 hr (r = 0.547 < 0.8), which means that there is a moderate relationship between the reduction of Tuta absoluta level of infestation at 24 and 48 hr with a significant difference (P = 0.037 < 0.05). The decrease of the level of infestation by Tuta absoluta is greater at 48 hr than its decrease at 24 hr after the treatment.
In addition, there is no significant difference between the level of infestation by Tuta absoluta at 48 and 72 hr after the treatment, with a mean of infestation equal to 0.91±0.22 and 1.08±0.23, respectively, and a very strong correlation and statistical relation between these two times (r = 0.880) (Table2).
ANOVA test showed that there is a highly significant difference between the efficiency of the tested groups at 48 hr and 72 hr after the treatment, F = 28.68, P = 0.000; F = 42.50, P = 0.000 respectively.  Table 2 shows that NOVOSECT SC21 ® was the best in controlling the emergence of new Tuta absoluta symptoms on the whole plant at 72 hours after the treatment (1.138±1.27) followed by ATO BED BUG ® and NEO-BOOST ® respectively (1.63±1.05; 1.43±1.07). The Duncan test showed similar results.

Effect of Biopesticides on the Larval Mortality of Tuta absoluta
The T-test between 24 and 48 hr after the treatment shows that the mean of the level of Tuta absoluta mortality/leaf are 0.71±0.15 and 0.7±0.22 respectively, with a very strong correlation value (0.949), and no significant difference (Table 3).
The ANOVA test showed that there is a highly significant difference between groups among time: at 24 hr F = 30.59, P = 0.000; at 48 hr F = 41.1, P = 0.000; at 72 hr F = 563.35, P = 0.000). NOVOSECT SC21 ® showed to be the most effective with a mortality level reaching 100% at 72 hr after the treatment. While ATO BED BUG ® has been more effective than NEO-BOOST ® among time. Similar results were also obtained using the Duncan test.

Effect of Biopesticides on Liriomyza trifolii Infestation
The T-test between the level of infestation by Liriomyza trifolii after 24 and 48 hr of the treatment, showed that the mean of the level of infestation by Liriomyza/plant is equal to 0.65±0.065, and 0.93±0.13 at 24 and 48 hr after the treatment, respectively. The correlation between the level of infestation/plant at 24 and 48 hr after the treatment is weak (0.369 < 0.8), which mean that there is a significant difference between the level of infestation by Liriomyza trifolii after 24 and 48 hr from the treatment. While the T-test between the level of infestation of Liriomyza trifolii 48 and 72 hr after the treatment is respectively 0.93±0.132 and 0.58±0.11, with a highly significant difference between the level of infestation by Liriomyza trifolii after 48 and 72 hr ( Table 4).
The ANOVA test showed that there is no significant difference among groups at 24 and 48 hours after the infestation: at 24 hr F = 1.149, P = 0.387; while at 48 hr F = 3.479, P = 0.07.

Effect of Biopesticides on Liriomyza trifolii Larval Mortality
The T-test between 24 and 48 hr after the treatment showed that the mean of Liriomyza trifolii dead larvae/leaf has decreased 0.84±0.14 and 0.71±0.23 respectively with a high correlation value (0.889) indicating a strong relation between the level of mortality after 24 and 48 hr from the treatment, with no significant difference among time (0.32 > 0.05) ( Table 5).
ANOVA test showed that there is a highly significant difference between groups at 24 and 48 hr after the treatment: at 24 hr F = 1.149±0.387; at 48 hr F = 3.479±0.070.

Discussion
Biopesticides such as clove oil (eugenol), BT F.D. 777 are excellent alternatives to synthetic insecticides as a means to reduce residues and protect the environment (Isman & Machal, 2006). The objective of our research was to evaluate the effect of ATO BED BUG ® , NOVOSECT SC21 ® and NEO-BOOST ® on the level of infestation and mortality of Tuta absoluta, Liriomyza trifolii and Alternaria solani.

Effect of Biopesticides on the Level of Tuta absoluta Infestation
The results showed that the effect of the treatments was completely opposite from the control: the resistance level of Tuta absoluta population decreased with time showing the best results of efficiency at 48 hr after the treatment. At 24 hr, biopesticides and the control showed same effect, while at 48 hr after the treatment, the three biopesticides (ATO BED BUG ® , NOVOSECT SC21 ® and NEO-BOOST) had the same opposite effect as the control. While at 72 hr after the treatment, NOVOSECT SC21 ® was the most efficient on reducing the level of infestation by Tuta absoluta, followed ATO BED BUG ® and NEO-BOOST ® . Through time, ATO BED BUG ® was the most efficient in reducing the infestation level and its activity was optimal after 48 hr from the treatment, time needed for the larvae of Tuta absoluta to ingest NOVOSECT SC21 ® metabolites. These results agree with the results of Derballa et al. (2012), who reported that the Bt metabolites having tpotential insecticidal activity against Lepidopterans pests. ATO BED BUG was also effective in reducing the infestation level by Tuta absoluta, but its efficacy decreased with time due to its volatility. Similar results were observed by Ebadah et al. (2006) showing a reduction of Tuta absoluta infestation between 50-60% when treated with clove oil under semi field conditions. Similarly, Mouawad et al. (2013) recorded that clove oil caused highly reduction percentage of penetration and accumulative mortality of larvae and caused ovipositional deterrence reaction towards adult stage of T. absoluta under laboratory conditions.

Toxicity of Biopesticides on Larvae of Tuta absoluta
NOVOSECT SC21 ® was the most effective in controlling T. absoluta by causing 92.40% mortality at 48 hr after the treatment: the larvae ingest the crystal inclusions (Cry toxins) which will be dissolved and activated in the alkaline environment of the insect gut, where it binds to specific receptors and cause the lyse of larvae midgut (Bravo et al., 2005). Similar observation was obtained with Derbalah et al. (2012) where a combination of Bacillus thuringiensis filtrate and Indoxacarb, have showed a reduction of larvae and mine blotch count in treated plants, and where Bt filtrates has exhibited satisfactory effectiveness against T. absoluta in greenhouses. ATO BED BUG ® was effective in increasing the mortality of T. absoluta larvae (82.77% at 48 hr after the treatment), similarly, Mouawad et al. (2013) affirmed that clove oil gave satisfactory results against T. absoluta.

Effect of Biopesticides on the Level of Liriomyza trifolii Infestation
The effect of all the biopesticides was optimal at 48 hr after the treatment. Similar results were reported by Ebadah et al. (2016) where the efficiency of three recommended insecticides (Acetamiprid 20% SP, Chlorpyrifos 48% and Lambda-cyhalothrin 5% EC) were compared to two natural oils (clove oil and bitter orange) against some of tomato insects (Liriomyza trifolii, Tuta absoluta and Bemisia argentifolii) under semi field conditions. The study showed a 48.7% reduction in tunnels after being treated with clove oil by between 3 and 5 days compared to a 52.5% reduction of tunnels after being treated with chlorpyriphos. Ebadah et al. (2016) reported that clove oil caused 56.8% mortality of L. trifolii compared to 59.2% and 69.1% mortality when treated with Chlorpyrifos and Lambda-cyhalothrin, respectively after three days. While after 5 days of the treatment, the mortality percent was increased to 61.4% when treated with clove oil compared to 43.7% by Acetamiprid and 40.1% by bitter orange. Based on the 215 day mortality mean, clove oil was the produced the greatest mortality of L. trifolii (60.5%). These results are in agreement with those obtained by Sabbour and Abd-El-Aziz (2010), who reported that clove oil and mustard revealed a strong repellent activity after 7 days (71 and 89%, respectively) against Bruchidius incarnates. Clove oil was also effective against other Dipteran's insects, such as Anopheles dirus mosquitoes (Trongtokit et al., 2005) and clove oil displays an insecticidal activity against this mosquito (Chaeib et al., 2007a). Cikman et al. (2006) reported that Bt is effective in controlling the larvae of L. trifolii and should be treated once every 2-3 weeks for effective control. According to Cikman (2006), Bt treated leaves have significantly fewer live larvae than non-treated leaves and that during the whole production period (15 weeks) the number of live larvae was still fewer than the number at which treatment is recommended (4-5 larvae per leaf). Ramirez-Godoy et al. (2018) showed that soil and foliar silicate applications enhanced plant resistance against the Asian Citrus Psyllid in Tahiti lime. Our results suggest that foliar NEO-BOOST applications should be considered by growers because it has an impact on Liriomyza populations.

Effect of Biopesticides on Alternaria solani Infection
ATO BED BUG ® was efficient in controlling Alternaria solani. According to Manohar et al. (2001), the phenolic components of clove oil: eugenol and carvacrol possess fungicidal characteristics on the cellular membrane. Also, clove oil has shown an antifungal activity against Candida albicans and Trichophyton mentagrophytes (Tampieri et al., 2005). Other studies have shown that a mixture of clove with concentrated sugar solution produced a strong fungicidal effect by reducing the fungi inoculum size (Nunez et al., 2001). Also, Pawar et al. (2006) affirmed that clove oil inhibited the growth of Asparagillus niger. The activity of clove oil against fungi is exerted on the cellular membrane and depends on the presence of aromatic ring and the presence of free phenol hydroxyl group and the lipophilic features of the components present in the oil (Tampieri et al., 2005;Cox et al., 2001;Chaeib et al., 2007b).

Conclusion and Recommendation
Our research indicates that NOVOSECT SC21 ® and ATO BED BUG ® has potential for controlling Tuta absoluta and Liriomyza trifolii by reducing the infestation level and increasing the larval mortality. Additional formulation research should be conducted to optimize the benefits of each material. This formulation research should involve investigating the impact of surfactants or other additives of the efficacy of the biopesticides and maximize their effective longevity once applied to the plants. Biopesticides might compete and replace conventional pesticides for the control of tomato leafminer Tuta absoluta, Lyromyza trifolii and Alternaria solani fungus. An economical study should be conducted in order to provide a better understanding of these biopesticides as alternatives to the conventional chemicals used by Hrajel regional farmers. Thus, these biopesticides are highly recommended to be implemented with other types of control measures as biological control agents or other biorationals in sustainable agro-ecosystems, such as organic farming and in integrated pest management (IPM) programs for Tuta absoluta, Liriomyza trifolii and Alternaria solani.