The Effect of Pyrethrum and Bacillus thuringiensis Biopesticides on Diprion pini L. and Neodiprion sertifer (Geoffr.) (Hymenoptera: Diprionidae) Larvae

Temel Göktürk1,* and Göksel Tozlu2

1Artvin Coruh University, Forest Faculty, Department of Forest Engineering, Artvin, Turkey

2Atatürk University, Faculty of Agriculture, Department of Plant Protection, Erzurum, Turkey

ABSTRACT

The larvae of the common pine sawfly (Diprion pini L.) and the European pine sawfly (Neodiprion sertifer) can cause epidemic, around the globe. Recently, an outbreak has been reported in young Scots pine (Pinus sylvestris L.) forest in Artvin - Borçka, Turkey. In order to propose effective control measures, this study was conducted to estimate the effectiveness of two biopesticides against D. pini ile N. sertifer larvae. Different doses of Pyrethrum and Bacillus thuringiensis biopesticides (Spruzit® Neu from 150ml to 600ml/100 l, and DiPel® DF BT 100 to 500 g/100 l) were applied against the larvae of D. pini and N. sertifer their impact was monitored under laboratory conditions. The finding of the study revealed that the most effective dosages for larvae of D. pini was Dipel 300 g/100 l and Dipel 500 g/100 l of B. thuringiensis. The efficiency ratio of Pyrethrum doses ranged from 59.5%-78.5% for Spunizet Neu and 85.5%-95.5% for Dipel DF against D. pini larvae. It was observed that the dosage of both pesticides was directly propositional to the death in larvae. Intriguingly, Dipel at the dose rate of 500 g/100 l was the most effective applications for larvae of N. sertifer. While variable impacts were noticed against larvae, both biopesticides were effective against larvae of D. pini and N. sertifer. Taken together, finding of this study propose the use of Pyrethrum and Bacillus thuringiensis biopesticides to control common pine sawfly and the European pine sawfly in the event of an epidemics in Turkey.

Article Information

Received 02 June 2018

Revised 11 July 2018

Accepted 22 July 2018

Available online 28 March 2019

Authors’ Contribution

TG and GT designed the study, each authors contributed equally to the study.

Key words

Diprion pini, Neodiprion sertifer, Pyrethrum, Dipel DF.

DOI: http://dx.doi.org/10.17582/journal.pjz/2019.51.3.989.994

* Corresponding author: [email protected]

0030-9923/2019/0003-0989 $ 9.00/0

Copyright 2019 Zoological Society of Pakistan

Introductıon

Turkey is an ecogeographically rich country, and 27% (20,712,894 ha) of the country-area is covered by the forests. Eastern Black Sea Region and Artvin are enriched with the natural forest and are represented by 54% and 2% of the Turkish forests, respectively. The dominant tree in the forests is Oriental Spruce, which is spread in an area of 25.628 ha. Other trees including Scotch pine, oriental beech, Abies nordmanniana (Steven) Spach, mountain alder, Anatolian chestnut and stone pine covers a significant proportion of the forest area (Eminağaoğlu et al., 2015). However, this rich forest is predisposed to several predators. Scotch pine (Pinus sylvestris L.) that represents an area of 217,104 hectares (5.37%) is under threat of reddish-yellow bush antenna sawfly (Neodiprion sertifer (Geoff.)) and bush antenna pine sawfly species (Diprion pini Linnaeus, 1758) (Hymenoptera: Diprionidae).

N. sertifer and D. pini are among the well-known coniferous tree pesticides for pine forest in the Europe (Barre et al., 2002; Dajoz, 2000; Turrisi and Bella, 1999). Additionally, these are reported in pine forests in Northern and Middle Europe and Asia and Northern America (Olofsson, 1987). These species cause epidemic periodically in prevalent areas leading to severe economic losses (Geri, 1988). In Turkey, both N. sertifer and D. pini species are predominantly prevalent in Mediterranean, Aegean, Marmara and Black Sea Regions, and cause damage to all pine forests (Çanakçıoğlu and Mol, 1998; Nafisi, 1999; Çuhadar et al., 2000; Şimşek and Kondur, 2006; Yaman et al., 2001; Aksu, 2010).

The damage caused by N. sertifer and D. pini is associated with eating leaves of pine trees. These damages are primarily reported in trees that are 10-15 years old. The trees attacked by N. sertifer and D. pini may become naked after all of their leaves are eaten and may be vulnerable to the attack of other pests because of their pre-existing weaknesses (Çanakçıoğlu and Mol, 1998; Romanyk and Cadahía, 2003; Augustaitis, 2007).

Today, silvicultural, mechanical, biological, biotechnical and chemical methods are being used as pests control strategies in forests. In Turkey, there are limited studies that detailed the extent of damage caused by D. pini and N. sertifer and mechanisms of control (Aksu, 2010; Akıncı and Avcı, 2016). Several methods are proposed to fight against these harmful pests, however, chemical control is one of the most frequently used methods around the globe (Demirbag et al., 1997). In Turkey, powder or liquid formulation of pesticides are recommended against these important sawflies (Çanakçıoğlu and Mol, 1998; Linstedt et al., 2006). There are several studies in which Diflubenzuron WP-25 was used against N. sertifer larvae in black pine plantation (Şimşek and Kondur, 2006). Additionally, it has been reported that decrease in pest population was observed after the use of the nuclear polyhedrosis virus (NsNPV) application against N. sertifer (Lord, 2005). Again, Anderbrant et al. (1998, 2000) and Östrand et al. (2000) reported that gonad pheromones could be used to reduce the N. sertifer populations and to monitor the population density.

According to the Environmental Protection Agency of the USA (EPA, 2014), there are more than 21,000 pesticides in the USA. Alternative biopesticides have been used for 30 years to synthetic insecticides and the production of synthetic chemicals has reduced at a rate of 2%, whereas the production of biopesticides has increased at a rate of 20% on annual bases (Cheng et al., 2010). Since chemical pesticides pose an environmental problem (Arora et al., 2012; Gülhane et al., 2015), integrated pest management (IPM), which includes biotechnical, mechanical and biological fights has become necessity rather than using the existing plant protection practices. The importance of biological fight, which is included in IPM, has increased in recent years. Applying entomopathogen organisms in biological fight has an important contribution in pest management. Among these organisms, it has been proven in many studies that Bacillus thuringiensis was effective against many pests (Martin and Bonneau, 2006; Cranshaw, 2008; Shaukat et al., 2010). Successful results were reported in studies in which many bacterial and biological agents were used against N. sertifer and D. pini, and these were recommended in the fight against pests (Mohamed et al., 1982; Inmaculata et al., 2001; Kees and Amanda, 2013; Van Frankenhyzen and Tonon, 2013). B. thrugiensis has been applied previously against D. pini in the fight in Artvin for trial purposes, and it was emphasized that this could be successful (Aksu, 2010). Plant-based insecticides have been used together with the organic agriculture practices. The most well known among these are azadirachtin, pyrethrum, rotenone, nicotine, ryania, sabadilla, quassine and plant oils (Güncan and Durmuşoğlu, 2004).

This study was designed to investigate the impact of pyrethrum and B. thuringiensis biopesticides on N. sertifer and D. pini pests and to assess the applicability for future control of these pests in the country.

 

MaterIals and methods

This study was conducted in the Scotch pine forest areas of Artvin Regional Forestry Directorate and Borçka Forestry Operation Directorate during 2016-2017. Both N. sertifer and D. pini larvae were collected from fresh Scotch pine shoots using hand pump and placed in tulle cages, before application of biopesticides. Different doses of biopesticides were applied against the larvae during in vitro conditions (Table I).

The 2nd and 3rd instar larvae of D. pini were collected in the 1st week of May, and the 2nd and 3rd instar larvae of N. sertifer were collected together with the branches of the young Scotch pine trees from which they fed on in the 2nd week of April. These samples were then brought to the Forest Entomology Laboratory in Forest Engineering Department of the Faculty of Forest at Artvin Çoruh University. The larvae that were brought with branches of the trees were placed in wire mesh cages with a size of 20x20x30cm as 20 larvae in each cage. The Scotch pine branches were submerged in moist flower turf to ensure that they remained humid. Biopesticides were applied at a pre-defined dosage (Table I) and monitored at every 12 hours to count and note the dead controls. The numerical data obtained in this process were evaluated with SPSS 15.0 package program. For the purpose of determining the effect of the biopesticides and their dosages on the death of N. sertifer and D. pini larvae, the One-Way variance analysis (ANOVA) was applied with numerical data. Using the Duncan Multiple Comparison Test we determined the most effective pesticide(s).

 

Table I.- Sampling organization.

Trade name of the pesticide	Dose	No of cages	No of larvae/cages
Pyrethrum Spruzit Neu	150ml/100 lt	10	20
	300ml/100 lt	10	20
	600ml/100 lt	10	20
Bacillus thuringiensis Dipel DF	100g/100 lt	10	20
	300g/100 lt	10	20
	500g/100 lt	10	20

 

Results

Assessing the impact of pesticides on pests revealed that all dosages applied in this study carried an inhibitory effect but at various levels. However, we noticed differences among six different slides applied to N. sertifer and D. pini larvae (Table II).

 

Table II.- One-way ANOVA results showing the effects of the pesticides and the doses applied on the larvae of N. sertifer and D. pini.

Period of development	Degree of freedom (df)	F value	Significance level (p)
N. sertifer larvae	5	72.38	0.001
D. pini larvae	5	70.12	0.001

 

Impact of biopesticide applications against the D. pini and N. sertifer larvae showed that pyrethrum had the same effect with as low as 150 ml/100 dose and as high as 300 ml/100 l dose. In the trial experiment, the efficiency of Dipel at 100 g/100 dose and pyrethrum at 600 ml/100 l dose showed comparable results. In laboratory experiment, all doses of Dipel, which had B. thuringiensis, showed the similar effects. The most effective death rates needed for the larvae of both pests to which biopesticide was applied was B. thuringiensis (Dipel DF 100 g/100 l), Dipel DF 300 g/100 l, Dipel DF 500 g/100 l dose applications (Table III).

The insecticidal effect of the biopesticides was assessed against D. pini larvae. It was found that pyrethrum (Spruzit Neu) was effective at 150 ml /100 l (59.5%); at 300 ml/100 l (62%); at 600 ml/100 l (78.5%) while Dipel DF was effective at 100 g/100 l (85.5%); at 300 g/100 l (93.5%); at 500 g/100 l (95.5%), respectively (Fig. 1).

The insectisidal effect of biopesticides for N. sertifer larvae was pyrethrum (Spruzit Neu) at 150 ml/100 l (55.5%) and followed by at 300 ml/100 l ( 66%), at 600 ml/100 l dose, at a ( 85.5%). Dipel DF was effected at 100 g/100 l ( 75%); at 300 g/100 l ( 86.5%); at 500 g/100 l ( 95%) (Fig. 2). The highest larva mortality was observed on the 4th and 6th days after the pesticide application in both species.

 

Table III.- Effects of the pesticides and the doses applied on the larvae and adults of N. sertifer and D. pini (Duncan Multiple Comparison Test p=0.05). Values are Mean±SD.

Trade name of the pesticide	Dose	N. sertifer	D. pini
Pyrethrum (Spruzit Neu)	150ml/100 lt	11.9±1.0 c	11.1±1.0 d
	300ml/100 lt	12.4±1.1 c	13.2±1.3 c
	600ml/100 lt	15.7±1.3 b	17.1±0.8 b
Bacillus thuringiensis (Dipel DF)	100g/100 lt	17.1±0.9 b,a	15.0±1.1 c
	300g/100 lt	18.7±0.7 a	17.3±0.9 b
	500g/100 lt	19.1±0.6 a	19.0±0.8 a

 

Discussion

Previous investigations have highlight that different biopesticides have varied intensities against N. sertifer in in-vitro. While the lethal effect of B. thuringiensis against N. sertifer larvae was found with 75-95% in the current study. This rate has been proposed as 35-71% by Inmaculata et al. (2001) and as 20.7% by Van Frankeyhzen (2009). While the lethal effect of B. thuringiensis against

 

 

D. pini larvae was found to be 85.5-95.5%; this rate was determined as 40% by Porcar et al. (2008); as 34-80% by Dadaşoğlu et al. (2016) and as 15-80% by Van Frankeyhzen and Tonon (2013). These differences could depend upon the genetic diversity of pests or biopesticides compositions. Nevertheless, all previous studies have proven the effectiveness of B. thuringiensis against these two pests and some different pests (Göktürk et al., 2018).

There is paucity of information indicating the lethal effects of plant-based insecticides on bugs and so far no studies have been conducted on the larvae of N. sertifer and D. pini. The lethal effect of pyrethrum was determined as 55.5-78.5%; the effect of pyrethrum on Pristiphora abietina (Christ, 1791) (Hymenoptera: Tenthredinidae) larvae has been investigated in a study conducted by Göktürk (2017) where it was determined as 71.7-98.8%. In present study, we observed that the dose of the pesticide is directly proportional to the death rates in larvae, a trend which has been be proposed earlier (Van Frankenhyzen and Gringorten, 1991).

It is now inevitable that harmful pests cause epidemic periodically in forest and agriculture areas leading to economic losses in crops. Although they do not cause direct deaths of trees, the damage caused by the N. sertifer and D. pini generally predispose trees to biotic and abiotic harmful factors. It is therefore imperative to design effective control strategies to safeguard the forests and secure the climate changes. The proposed study investigates the pyrethrum (Spruzit® Neu) and B. thuringiensis (DiPel® DF) biopesticides against N. sertifer and D. pini larvae in laboratory conditions. Results their effectiveness against both tested pests even at low doses, which underline the economics and affordability of the farmers. It was also observed that the death rates increased especially on the 4th and 6th days post-pesticide applications highlighting the extent of responsiveness.

 

Conclusion

Based on these finding, it can be concluded that Pyrethrum and B. thuringiensis biopesticides should be applied against N. sertifer and D. pini larvae in the field conditions. In case successful results are achieved in in-vivo conditions, it will have great importance to transfer them to use in the fight against pests.

 

Statement of conflict of interest

Authors have declared no conflict of interest.

 

References

Akıncı, Z.E. and Avcı, M., 2016. Biology and natural enemies of Neodiprion sertifer in the Lakes District forests. Turk. J. Forest., 17: 30-36.

Aksu, Y., 2010. Investigation on Neodiprion sertifer (Geoff.) (Hymenoptera; Diprionidae) which has important damages in Pinus sylvestris in afforestation areas. J. Forest Engin., 47: 26-34.

Anderbrant, O., Högberg, E., Hedenström, E. and Löfqvist, J., 1998. Towards the use pine sawfly pheromones in forest protection: Evaluation of a behavioral antagonist for mating disruption of Neodiprion sertifer. In: Proceedings: Population dynamics, impacts, and integrated management of forest defoliating insects (eds. M.L. McManus and A.M. Liebhold). USDA Forest Service General Technical Report NE-247, pp. 53-63.

Anderbrant, O., Löfqvist, J., Högberg, H. E., Hedenström, E., Baldassari, N., Baranio, P., Kolmakova, G., Lyons, B., Naito, T., Odinokov, V., Simandl, J., Supatashvili, A., Tai, A. and Tourianov, R., 2000. Geographic variation in the field response of male European pine sawflies, Neodiprion sertifer, to different pheromone stereoisomers and esters. Ent. Exp. Appl., 95: 229-239. https://doi.org/10.1046/j.1570-7458.2000.00662.x

Arora, N.K., Tewari, S., Singh, S., Lal, N. and Maheshwari, D.K., 2012. PGPR for protection of plant health under saline conditions. In: Bacteria in agrobiology: Stress management (ed. D.K. Maheshwari). Springer, Berlin, pp. 239-258. https://doi.org/10.1007/978-3-662-45795-5_12

Augustaitis, A., 2007. Pine sawfly (Diprion pini L.) – related changes in Scots pine crown defoliation and possibilities of recovery. Pol. J. Environ., 16: 363-369.

Barre, F., Milsant, F., Palasse, C., Prigent, V., Goussard, F. and Geri, C., 2002. Preference and performance of the sawfly Diprion pini on host and non-host plants of the genus Pinus. Ent. Exp. Appl., 102: 229-237. https://doi.org/10.1046/j.1570-7458.2002.00944.x

Çanakçıoğlu, H. and Mol, T., 1998. Forest entomology: Harmful and useful insects. Istanbul University Forestry Faculty Publications, IX+541s.

Cheng, X.L., Liu, C.J. and Yao, J.W., 2010. The current status, development trend and strategy of the bio-pesticide industry in China. Hubei Agric. Sci., 49: 2287-2290.

Cranshaw, W.S., 2008. Bacillus thuringiensis. Colorado State University Extension No. 5, pp. 556.

Çuhadar, A.G.I., Aksu, Y. and Babacan, N., 2000. Investigations on Neodiprion sertifer (Geoff.) damaged in forests in Artvin Region. J. Forest Engin., 37: 18-24.

Dadasoglu, F., Tozlu, G., Kotan, R., Gokturk, T. and Kenan, K., 2016. Biological control of pine sawfly (Diprion pini L.) and molecular characterisation of effective strains. Romanian Biotechnol. Lett., 21: 2.

Dajoz, R., 2000. Insects and forests. The role of diversity of insects in the forest environment. Intercept Ltd., London.

Dajoz, R., 2001. Forest Entomology: insects and the forest. Mundi-Press Editions, Madrid.

Demirbag, Z. and Beldüz, A.O., 1997. The Importance of Baculovirus in biological control. Kükem Derg., 20: 49-58.

Eminağaoğlu, Ö., Akyıldırım, H. and Aksu, G., 2015. Artvin’s Natural Plants. Promat, Istanbul, pp. 27-52.

EPA, 2014. Pesticides: Regulating pesticide. Environmental Protection Agency, U.S.A. http://www.epa.gov

Ge´ri, C., 1988. The pine sawfly in central France. In: Dynamics of forest insect populations: Patterns, causes, implications (ed. A.A. Berryman). Plenum Press, New York, pp. 377-405. https://doi.org/10.1007/978-1-4899-0789-9_19

Göktürk, T., 2017. The effect of Pyrethrum and Bacillus thuringiensis against Pristiphora abietina (Christ, 1791) (Hymenoptera: Tenthredinidae). Artvin Coruh Uni. J. Forest. Fac., 18: 83-87.

Göktürk, T., Tozlu, E. and Kotan, R., 2018. Prospects of entomopathogenic bacteria and fungi for biological control of Ricania simulans (Walker 1851) (Hemiptera: Ricaniidae). Pakistan J. Zool., 50: 75-82, 2018. DOI: http://dx.doi.org/10.17582/journal.pjz/2018.50.1.75.82

Gulhane, P.A., Gomashe, A.V. and Sundarkar, K.M., 2015. Influence of pesticides on nitrogen fixing bacteria. Int. J. Tech. Res. Applic., 3: 157-160.

Güncan, A. and Durmuşoğlu, E., 2004. An evaluation on natural insecticides with herbal origin. Hasad Dergisi, 20: 26-32.

Inmaculada, G.R., Sa´nchez, J., Gruppe, A., Martı´nez-Ramı´rez, A.C., Rausell, C., Real, M.D. and Bravo, A., 2001. Mode of action of Bacillus thuringiensis PS86Q3 strain in hymenopteran forest pests. Insect Biochem. mol. Biol., 31: 849-856. https://doi.org/10.1016/S0965-1748(01)00030-3

Keesvan, F. and Amanda, T., 2013. Activity of Bacillus thuringiensis cyt1Ba crystal protein against hymenopteran forest pests. J. Inverteb. Pathol., 113: 160-162. https://doi.org/10.1016/j.jip.2013.03.007

Lindstedt, C., Mappes, J., Päivinen, J. and Varama, M., 2006. Effects of group size and pine defence chemicals on Diprionid sawfly survival against ant predation. Oecologia, 150: 519-526. https://doi.org/10.1007/s00442-006-0572-3

Lord, J.C., 2005. From Metchnikoff to Monsanto and beyond: The path of microbial control. J. Inverteb. Pathol., 89: 19-29. https://doi.org/10.1016/j.jip.2005.04.006

Martin, J.C. and Bonneau, X., 2006. Bacillus thuringiensis, 30 years of control of cluster caterpillars. Phytoma, 590: 4-7.

Mohamed, M.A., Coppel, H.C. and Podgwaite, J.D., 1982. Persistence in soil and on foliage of nucleopolyhedrosis virus of the European pine sawfly, Neodiprion sertifer (Hymenoptera: Diprionidae). Environ. Ent., 11: 1116-1118. https://doi.org/10.1093/ee/11.5.1116

Nafisi, S., 1999. Determination of harmful insect species and their biology in Pinus brutia (Tenora) in Karabuk Forest Operation Directorate, Kaplan afforestation areas. PhD Thesis, Zonguldak Karaelmas University, Graduate School of Natural and Applied Sciences Department of Forest Engineering, Bartın, Turkey. pp. 134.

Olofsson, E., 1987. Mortality factors in a population of Neodiprion sertifer (Hymenoptera: Diprionidae). Oikos, 48: 297-303. https://doi.org/10.2307/3565517

Östrand, F., Anderbrant, O. and Jönsson, P., 2000. Behavior of male pine sawflies, Neodiprion sertifer, released downwind from pheromone sources. Ent. Exp. Appl., 95: 119-128. https://doi.org/10.1046/j.1570-7458.2000.00649.x

Porcar, M., Go´mez F., Gruppe A., Go´mez-Pajuelo A, Segura I. and Schroder, R., 2008. Hymenopteran specificity of Bacillus thuringiensis strain PS86Q3. Biol. Contr., 45: 427-432. https://doi.org/10.1016/j.biocontrol.2008.02.002

Romanyk, D. and Cadahía, D., 2003. Insect pests in the Spanish forest stands, 4th ed. Ministerio de Medio Ambiente, Madrid.

Shaukat, A., Yusuf, Z., Ghulam Muhammad, A. and Farhat, N., 2010. Bacillus thuringiensis and its application in agriculture. Afri. J. Biotechnol., 9: 2022-2031.

Şimşek, Z. and Kondur, Y., 2006. Bacillus thuringiensis and its application in agriculture. Kastamonu Uni.J. Forest.Facul., 6: 105-107.

Turrisi, G. and Bella, S., 1999. First reporting of Diprionidae for the Sicilian fauna (Hymenoptera: Synphyta). Bull. Soc. Ent. Ital., 131: 179-182.

Van Frankenhuyzen, K., 2009. Insecticidal activity of Bacillus thuringiensis crystal proteins. J. Inverteb. Pathol., 101: 1-16. https://doi.org/10.1016/j.jip.2009.02.009

Van Frankenhuyzen, K. and Gringorten, L., 1991. Frass failure and pupation failure as quantal measurements of Bacillus thuringiensis toxicity to Lepidoptera. J. Inverteb. Pathol., 58: 465-467. https://doi.org/10.1016/0022-2011(91)90199-Z

Van Frankenhuyzen, K. and Tonon, A., 2013. Activity of Bacillus thuringiensis Cyt1Ba crystal protein against hymenopteran forest pests. J. Inverteb. Pathol., 113: 160-162. https://doi.org/10.1016/j.jip.2013.03.007

Yaman, M., Nalçacıoğlu, R. and Demirbağ, Z., 2001. Viral control of the European pine sawfly, Neodiprion sertifer (Geoffroy) in Turkey. Turk. J. Biol., 25: 419-425.