Integrated Control of Tropilaelaps clareae And Varroa destructor In Apis mellifera, L. Colonies

Transcription

1 Scholarly Journal of Agricultural Science Vol. 4(1), pp January, 2014 Available online at ISSN Scholarly-Journals Full Length Research Paper Integrated Control of Tropilaelaps clareae And Varroa destructor In Apis mellifera, L. Colonies *Rashid Mahmood, **Atta Ul Mohsin, **Saima Asad, *Shazia Raja, *Ghulam Sarwar *Honeybee Research Institute, National Agricultural Research Centre, Islamabad, Pakistan ** PMAS-Arid Agriculture University, Rawalpindi, Pakistan. Accepted 14 November 2013 Integrated Management was carried out to determine the effects of three different treatments groups. i.e. 4gm thymol + 3.2% OA and 65% formic acid (T 1 ), 5% clove oil + Tobacco extract and 4gm thymol+3.2% OA (T 2 ) and 5% clove oil + Tobacco extract and 65 % formic acid (T 3 ) for the control of ectoparasitic mites i.e. Tropilaelaps clareae and Varroa destructor populations in honeybee Apis mellifera colonies round the year at Honey bee Research Institute, NARC, Islamabad. Average mite mortality, efficacy and honey yield was calculated. It was found that T 1 had the highest mite mortality number 121, 724 and efficacy 86, % for T. clareae and V. destructor, respectively. The total honey production harvested from colonies treated with different acaricides was also determined and significantly more mean amount of honey was produced (30 kg) from the hives treated with 4gm thymol + 3.2% OA and 65% formic acid. It was observed that during all experiments treatment cause no effect upon queen and adult honey bee activities. Key words: - Formic acid, Oxalic acid, IPM, Ectoparasitic mites, honey. INTRODUCTION In Pakistan, beekeeping is a profitable business. It is reported that there are more than 4,000 beekeepers rearing Apis mellifera in the beehives, about 400,000 colonies of A. mellifera has been producing 10,000 MT honey annually and families are being benefited from beekeeping (Annual Report, PARC ). Pakistan Agricultural Research Council (PARC) in late introduced the western honeybee A. mellifera from Australia among the commercial beekeepers because least output of A. cerana (Waghchour and Martin, 2008). Just after the introduction of A. mellifera in Pakistan, V. destructor mite became a serious pest of this newly introduced A. mellifera and attacked over a large number of honeybee colonies (Ahmad, 1988). Ectoparasitic mites infestations results in low yield of honey, swarming and absconding of bee colonies. The two mite species V. destructor and T. clareae are considered as the cause of continued increasing *Corresponding author. rashid_ento1@ yahoo.com. infestation among the Apis mellifera colonies in Asia (De Jong et al., 1982). Each year a considerable damage among in bee colonies were caused by mite infestation. As a result, capital flight was observed among the beekeeping industry (Khan et al., 1987). Besides the economical loss of both honey bees and honey yield, it was expected; infested colony may migrate or die (Needham, 1988). Varroa mites growths depend on all honeybee stages from larva to adult bees. A large number of adult bees were found in front of bee hive attacked by mites. Colonies heavily infected by Varroa mite become unproductive (Ritter, 1981). The mite T. clareae is parasitic on bee brood which causes brood malformation, death of the bees and subsequent colony decline or absconding. Development requires about one week and mites are dispersed on bees. T. clareae are small mites and not easily seen or collected. This mite attacks both the adult and brood. T. clareae occurs on 5 species of bee - Apis cerana, A. dorsata, A. florea, A. laboriosa and A. mellifera. T. clareae is known to have a wide distribution in Asia extending eastwards from Iran to Papua New Guinea

2 Mahmood et al. 33 (Matheson, 1995).The greatest infestation occurred in February, March and April with a decline from May to August (Camphor et al., 2005). Poor management of bee colonies, hive microclimate, strength of the colony increased the prevalence of T. clareae in bee colonies (Mahavir and Gupta, 1999). T. clareae caused 30-70% colony loss of A. mellifera with reduced honey production (Woo and Lee, 1997). Presently, synthetic acaricides i.e. chlorobenzilate, sulphur, phenothiazine, amitraz or different pyrethroids can be used to control these mites. Different kinds of acaricides were used effectively to control mites infestation but with the passage of time mite population started getting resistant against many successful acaricides (Loglio and Plebani, 1992). While designing the present study an integrated management approach was taken into consideration for the control of mites. In this regard various concentrations of organic chemicals, clove oil (Syzygium aromaticum) and Tobacco (Nicotiana tabacum) extract will be tested individually and in various combinations around the year to evaluate their efficacy against mite populations of honeybees. As a result a set of new recommendations would be approached to be practiced by the beekeepers to manage the mite population and will improve the honey yield and indirectly will impact the livelihood and economic condition of the beekeepers. MATERIALS AND METHODS About 150 adult and sealed brood populations of Honeybee Research Institute apiaries were assessed for infestation before selecting the experimental colonies. To collect the sample of 250 bees/ colony for mite infestations level by using alcohol wash technique (De Jong et al., 1982). The mite infestation was also evaluated by opening 100 cells of sealed brood before treatment (Burgett and Burikam, 1985) and for the assessment of mite population level in debris mite collection trays kept for 24hrs in the bottom board of bee hive and mites was counted (Devlin, 2001). Finally, thirty queen right honeybee colonies in Langstroth hives were used on equal mite infestation levels with at least ten bee frames. The colonies were placed in the premises of Honeybee Research Institute, NARC, Islamabad at different locality of beekeeping areas. Each honeybee colony was equipped with a modified bottom board and a mite collection tray (mite excluder) which was placed through the back side of the hive, without disturbing colony. Efficacy of mite infestation was calculated by count of falling mites in debris. The honeybee colonies of each group were placed at appropriate distance of 5 meters. Colony strength (number of combs covered with bees, brood areas, and amount of food) were equal (Rashid et al., 2011). Colonies were divided into three groups of ten colonies each with the following treatments. One group (T1) = Three applications of 4 gm thymol % oxalic acid solution in the November-December, 2010 and two application of 20 ml formic acid (65%) in July,2011. Second group (T2) = Three applications of 5% clove oil (Syzygium aromaticum) + Tobacco (Nicotiana tabacum) extract in March, 2010 and two applications of 4gm thymol+3.2% OA solution in December, 2010 and January, Third group (T3) = Three applications of 5% clove oil + Tobacco extract in July, 2010 and two applications of 65 % formic acid in March, Treatments were applied every evening when the honeybees were present in the hives. Thymol crystals (finely grinded) were placed in Petri dishes (80mm dia) on top of the brood frame under the top cover of hives spaces that contained bees and 65% formic acid (20 ml) applied on each card board placed in the mite collection trays placed in the deep bottom board of the hive (Rashid et al., 2011). Oxalic acid was applied in sugar syrup. To obtain 3.2 % OA solution, 75 g oxalic acid dehydrate was mixed with 1 liter of sugar water (1:1) (Prandin et al., 2001). Treatments were only delivered to frame spaces that contained bees; any empty frame was not treated. The 5 ml mixture was trickled directly on the adult bees in between two frames using a syringe as recommended (Imdorf et al., 1997 and Brodsgaard et al., 1999). 15 milliliters of clove oil and tobacco extract was sprayed by using plastic sprayer (500 ml). Solutions of all clove oil and tobacco extract concentrations were sprinkled over the bees inside the colonies (Zaitoon, 2001). At the end; all the experimental colonies were given Fluvalinate (Apistan) strip for knockdown. Apistan strips were removed from the colonies after four weeks and dropped dead mites were counted (Marcangeli and Garcia, 2004). All the colonies were checked for dead worker bees and queens at the end of treatment application. The efficacy of the treatments was calculated by using following formula (Marinelli et al., 2004): HONEY YIELD Honey was harvested after all experiments with the help of manual honey harvester and compared honey yield of treated and control honeybee colonies. Honey (Acacia and ber) production was measured by taking the weight of each hive body used for honey collection before and after the honey extraction process. The weight difference was considered as the amount of harvestable honey (Rashid et al., 2011).

3 Scholarly J. Agric. Sci. 34 Figure 1. Mean number of mites fallen by various treatments for T. clareae and V. destructor bars STATISTICAL ANALYSIS All data collected by performing experiments were statistically being analyzed through MSTAT C computer based software (Freed and Eisensmith, 1986). Analysis of Variance was used at 5% probability level (Montgomery, 2001). RESULTS AND DISCUSSION The experiment shows that the number of Tropilaelaps mites fallen for the T1, T2 and T3 ranged between , and and in the case of Varroa the ranges were , and , respectively. The mean number of Tropilaelaps and Varroa fallen for each treatment T1, T2 and T3 were ± 1.19, 66.90± 1.89, 88.90± 2.30 and ± 4.47, 175± 2.09, 195± 3.18 respectively as shown in Figure 1. When different treatments were compared a highly significant difference was found for the number of fallen mites. The highest number of mites fell in T1 and when different treatments were compared it was found that only T1 was highly significantly different from all the other treatments, which clearly showed T1 to be the most effective against Tropilaelaps and Varroa mites (Table 1) at 5% level of significance. Results are confirmed by Rashid et al., 2011 who concluded that 65% Formic acid and thymol more effective for the control of T. clareae. Rashid et al., 2012a reported 3.2% OA concentration are very effective for the control of Varroa. Rashid et al., 2012b also confirmed 4gm thymol + 3.2% OA combination is effective for the control of both types of ectoparasitic mites in broodless condition. Amrine and Noel, 2007 concluded that 50% formic acid, seperamint, lemongrass, essential oils with use of screen bottom boards trays collectively effective for the control of Varroa mites. Harold et al. (1989) reported that 94% mites were killed by application of 4 treatements of formic acid and the most effective treatment (62% of mites killed) was with 40 ml of 65% formic acid (Greatti et al., 1993). Thymol effectual against mites but safe to honeybees (Bollhalder 1998 and Calderone 1999). Chiesa (1991) and Gal et al. (1992) Lensky et al. (1996) reported that 30% thymol was harmful to bee colonies during summer. Oxalic acid was found very effective for control of mites is in confirmation with the results showing that OA is very effective against V. destructor (Gregorc and Planinc 2001 and 2002; Gregorc and Poklular 2003; Marinelli et al., 2004). Imdorf et al., (1997) claimed that OA did not effect on brood area and Wachendorfer et al., (1985) observed no bee mortatlity, no loss of queen and no supersedure

4 Mahmood et al. 35 Table 1. Multiple Comparisons of different treatments for the control of mites Treatment Mite Mortality (#) Efficacy (%) Honey T. clareae V. destructor T. clareae V. destructor Yield (Kg) 4gm thymol + 3.2% OA a a a a a and 65% formic acid 5% clove oil+ tobacco extract 66.90c c b b b and 4gm thymol + 3.2% OA 5% clove oil+ tobacco 88.90b b c c b extract and 65 % formic acid LSD (0.05) F-test 214** 8458** 30.1** 494** 101** C.V **Highly significant at 0.1%. Figure 2. The mean efficacy (%) by various treatments for T. clareae and V. destructor bars. by the application of OA. Further in case of Tropilaelaps clareae the range of efficacy in colonies treated with T1, T2 and T3 were 84-88, and 68-83%, respectively. The mean value of efficacy 86± 0.37, 81± 066 and 75± 1.49 for the said treatments also varied between different treatments and when compared were found to be significantly different (at 5% level of significance) where T1 was found to have the highest efficacy. For the Varroa mites the efficacy range for T1, T2 and T3 was 97-98, and 81-86%. The mean value of efficacy for T1, T2 and T3 were 98± 0.74, 92± 0.23 and 83± 0.53 respectively. The results showed a highly significant difference between efficacies (at 5% level of significance). The T1 again showed the highest average efficacy of 98% (Figure 2, Table 1). Results confirmed by Rashid et al., 2011, 2012a,b that by the application of thymol, formic acid and oxalic acid have more effective for the control of ectoparasitic bee mites without any side effects. Three application of formic acid in bee hive shown

5 Scholarly J. Agric. Sci. 36 Figure 3. The mean amount of honey produced from colonies treated with different treatments good efficacy against mite control (Mutenelli et a1., 1994 and Van Veen et al., 1998) which is also confirmed by the experiment where 2-3 doses of formic acid were used. The honey produced from different hives when treated with different treatments was also weighed at the end of experiment. The range of honey in colonies treated with T1, T2, T3 was 28-33, and kg, respectively. The mean amount of honey produced in kg from T1, T2 and T3 is shown in Figure 3 and Table 1. Results showed a significantly more amount of honey (30 kilogram) from the colonies treated with T1 (at 5% level of significance). Results confirmed by Rashid et al., 2011, 2012a,b that by the application of thymol, formic acid and oxalic acid produced more honey without any side effects. Mattila and Otis (1999) showed that honey yield was reduced by 30% during the formic acid treatment. On contrast, Mutinelli et al. (1996) reported low bee mortality in all tests of formic acid and also El-Shaarawy (1999) found that honey yield increased when apiaries treated with formic acid. Less value of C.V. indicates high precision of the experiment in mite mortality, efficacy and honey yield in first group (4gm thymol + 3.2% OA and 65% formic acid) as compared to other groups (Table i). CONCLUSION It is concluded that 4gm thymol + 3.2% oxalic acid solution and 65% formic acid applied throughout the year along with maintaining regular re-queening and use of modified bottom board trays have shown 2 times in T. clareae and 4 times in V. destructor best results on mite mortality. Average honey yield increased up to 30 kilogram in T1 group treated colonies. RECOMMENDATIONS Based on research findings, it is recommended that regular re-queening with hygienic queen, use of modified bottom boards along with three applications of 4 gm thymol+3.2% Oxalic acid in mid winter at weekly intervals, two application of 65% formic acid in mid summer fortnightly and apistan strips should be hanged in late summer for a period of one month for successfully controlling both ectoparasitic mites (Tropilaelaps clareae and Varroa destructor) of Apis mellifera and for the production of quality honey yield. REFERENCES Ahmad, R. (1988). Honeybee parasitic mites and their control in Pakistan, Prog. Far., 8: Amrine, J.W., Noel, R. (2007). Formic Acid Fumigator for Controlling Varroa Mites in Honey Bee Hives, Int. J. Acarol., 32(2): Bollhalder, F. (1998). Thymovar for Varroa control, Schweizerische Bienen Zeitung., 121: Brodsgaard, C. J., Jensen, S. E., Hansen, C. W., Hansen, H. (1999). Spring treatment with oxalic acid in honeybee colonies as Varroa control, DIAS Report, Horticulture No.6, 2nd edition.pp16. Burgett, D.M., Burikam, I. (1985). Number of adult honeybees (Hymenoptera: Apidae) occupying a comb: A standard for estimating colony populations, J. Econ. Entomol., 78: Calderone, N.W. (1999). Evaluation of formic acid and a Thymol-based blend of natural products for the fall control of Varroa jacobsoni (Acari: Varroidae) in colonies of Apis mellifera (Hymenoptera: Apidae), J. Econ. Entomol., 92: Camphor, E.S.W., Hashmi, A.A., Writter, W., Bowen, I.D. (2005). Seasonal changes in mite (Tropilaelaps clareae) and honeybees (Apis mellifera) population in Apistan treated and untreated colonies, Apiacta., 40: Chiesa, F. (1991). Effective control of Varroatosis using powdered thymol, Apidol., 22 (2): Dejong, D., Deandrea, R.D., Goncalves, L.S. (1982). A comparative analysis of shaking solutions for the detection of Varroa jacobsoni on

6 Mahmood et al. 37 adult honeybees, Apidol., 13: Devlin, S. (2001). Comparative Analyses of sampling methods for Varroa mite (Varroa destructor Anderson and Trueman) in honey bees (Apis mellifera L.), MPM.Thesis, Simon Fraser Univ., Burnaby BC, Canada. El-Shaarawy, M. O. (1999). Evaluation of Apiguard and formic acid as control agents against Varroa jacobsoni infesting honeybee colonies, Proceed. Apimondia 99, Congress XXVIe, Vancouver Sept., Canada pp. Freed, R. D., Eisensmith, S.P. (1986). MSTAT Microcomputer Statistical program. Michigan State University of Agriculture and Applied Science, Michigan. USA. Gal, H., Slabezki, Y., Lensky.Y. (1992). A preliminary report on the effect of origanum oil and thymol applications in honeybee (Apis mellifera L.) colonies in a subtropical climate on population levels of Varroa jacobsoni, Bee Sci., 2 (4): Greattti, M., Bartbattimi, R.D., Agaro, M. (1993). Treatment of Varroasis; Formic acid as a control method against Varroa jacobsoni, Obiettivi e Documenti Veterinari., 14(6): Gregorc, A., Poklukar, J. (2003). Rotenone and oxalic acids as alternative acaricidal treatments for Varroa destructor in honey bee colonies, Vet.Parasito.,111: Gregore, A., Planinc, I. (2002). The control of Varroa destructor using oxalic acid, Vet. J., 163 (3): Gregore, A., Planinc, I. (2001). Acaricidal effect of oxalic acid in honeybee (Apis mellifera) colonies, Apidologie., 32(4): Harold, H., Wolfgang, R., Elizabeth, S.W. (1989). The control of parasitic bee mites: Varroa jacobsoni, acarapis woodi and Tropilaelaps clareae with formic acid, Am. Bee J., 129: Imdorf, A., Bogdanov, S., Ochoa, R.I., Calderone, N.W. (1999). Use of essential oils for the control of Varroa jacobsoni Oud. in honey bee colonies, Apidol., 30: Imdorf, A., Charriere, J. D., Bachofen, B. (1997). Efficiency checking of the Varroa jacobsoni control methods by means of oxalic acid, Apiacta., 32: Khan, B. M., Chaudhry, M.I., Khawaja, P. (1987). Acarine disease of honey bees and its control in Pakistan, Pak. J. Forest., 37: Lensky, Y., Slabezki, Y., Gal, H., Gerson, V., Dechmani, V., Jirasavetakul, V. (1996). Integrated control of Varroa mite (Varroa jacobsoni Oud.) in Apis mellifera colonies in Thailand,. Final report., Agric. Res. Agreement for Dev. Countries. 96 pp. Loglio, G., Plebani, G. (1992). Valutanione dell efficacia dell Apistan, Apis. Mod., 83: Mahavir, G., Gupta, M. (1999). Infestation of Apis mellifera L. colonies with ectoparasitic mites, Ann. Biol., Ludhiana 15(2): Marcangeli, J., Garcia, M. D. C. (2004). Effect of Apis mellifera (Apidae) honeybee brood amount on Oxavar acaricide efficacy against the mite Varroa destructor (Varroidae), Rev. Soc. Entomol. Agr., 63(3-4): Marinelli, E., Pulcini,P., Margio, C., De Pace, F., Allegrini, F., Persona, F., Oddo, L. (2004). Oxalic acid by Varrox to Varroa control in Central Italy, Apiacta., 39: Matheson, A. (1995). World bee health report, Bee World., 74: Mattila, H.R., Otis, G.W. (1999). Trials of Apiguard, thymol-based miticides. Part1. Efficacy for control of parasitic mites and residues in honey, Am. Bee J., 139 (12): Montgomery, D.C. (2001). Design and Analysis of Experiments, 5 th Ed. John Willy and Sons, New York. p Mutinelli, F., Cremasco, S., Irsra, A. (1994). Formic acid in the control of Varroatosis: a practical approach, J. Vet. Med., 41: Mutinelli, F., Cremasco, S., Isara, A., Baggio, A., Nanetti, A., Massi, S. (1996). Organic acids and Apilife VAR in the control of Varroosis in Italy, Apicoltore Moderno., 87(3): Needham, G. R. (1988). Status report on Varroa jacobsoni, Am. Bee J., 128: PARC. ( ). Honeybee,. Ann. Rep., PARC. P. 98. Prandin, L., Nicoletta, D., Barbara, G., Ornella, D., Roberto, P., Franco, M. (2001). A scientific note on long-term stability of a home-made oxalic acid water sugar solution for controlling varroosis, Apidol., 32: Rashid, M., Wagchoure, E. S., Moshin, A.U., Raja, S., Sarwar, G. (2012a). Control of ectoparasitic mite varroa destructor in honeybee (Apis mellifera) Colonies by using different concentrations of oxalic acid, J.Anim. Plant Sci., 22(1): Rashid, M., Wagchoure, E. S., Moshin, A.U., Raja, S., Sarwar, G. (2012b). Control of ectoparasitic mites in honeybee (Apis mellifera L.) colonies by using thymol and Oxalic acid, Pak. J. Zoo., 44(3): Rashid, M., Wagchoure, E. S., Raja, S., Sarwar, G., Aslam, M. (2011). Effect of thymol and formic acid against ectoparasitic brood mite Tropilaelaps clareae in Apis mellifera colonies, Pak. J. Zoo., 43(1): Ritter, W. (1981). Varroa disease of honeybee Apis mellifera, Bee world., 62(4): Van Veen, J., Caledron Fallas, A., Cubero Murillo, A., Arce, H. (1998). Varroa jacobsoni in Costa Rica: detection, spread, and treatment with Formic acid, Bee World.,79:5-10. Wachendorfer, G., Fijalkowski, J., Kaiser, E., Seinsche, D., Siebentritt, J. (1985). Labor und Feldversuche mit der Illertisser Milbenplatte als neue Anwendungsform der Ameisensäure im Rahmen der Varroatose-Bekämpfung. [Laboratory and field tests with the illertisser milbenplatte (mite plate) a new way of application of formic acid to control varroatosis.], Apidol., 16: Waghchour-Camphor, E.S., Martin, S. J. (2008). Beekeeping in Pakistan a bright future in a troubled land, Am. Bee J., 148: Woo, K.S., Lee, J.H. (1997). Current status of honeybee mites in Koressa,. Honeybee Sci., 18(4): Zaitoon, A.A. (2001). Evaluation of certain plant extracts for the control of parasitic bee mites, Varroa jacobsoni, J. Pest and environ. Sci., 9:77-88.