Our study was conducted with the permission of United States Sugar Corporation. We want to thank R. Raid, S. Raid and T. Lang for their assistant. We also M. Stanback and three anonymous reviewers for suggestions that improved this manuscript.

1. Construction of Swarm Trap Boxes
NOTE: There are two alternate protocols for swarm trap construction included as steps 1.1 and 1.2. There are several additional ways trap boxes could be constructed. The following two methods are presented here because we have tested them in experiments and they have been successful. For either protocol, use 1.3 cm to 2 cm thick non-pressure treated plywood.
<ol>
	<li>Construction of Swarm Trap Protocol Option 1: Building a Trap Box
	<ol>
		<li>Cut six pieces of plywood 30 cm ...

<ol>
	<li>Winston, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biology+and+management+of+Africanized+honey+bees.">The biology and management of Africanized honey bees.</a> Annu Rev Entomol. 37, 173-193 (1992).</li><li>Schmidt, J. O., Hurley, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selection+of+nest+cavities+by+Africanized+and+European+honey+bees.">Selection of nest cavities by Africanized and European honey bees.</a> Apidologie. 26, 467-475 (1995).</li><li>Eadie, J., Sherman, P., Semel, B., Casro, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conspecific+Brood+Parasitism,+Population+Dynamics,+and+the+Conservation+of+Cavity-Nesting+Birds.">Conspecific Brood Parasitism, Population Dynamics, and the Conservation of Cavity-Nesting Birds.</a> In: Behavioral Ecology and Conservation Biology. , 306-340 (1998).</li><li>Newton, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+nest+sites+in+limiting+the+numbers+of+hole-nesting+birds:+a+review.">The role of nest sites in limiting the numbers of hole-nesting birds: a review.</a> Biol Conserv. 70, 265-276 (1994).</li><li>White, T. H., Abreu-Gonz&#225;lez, W., Toledo-Gonz&#225;lez, M., Torres-B&#225;ez, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+the+field:+artificial+nest+cavities+for+Amazona+parrots.">From the field: artificial nest cavities for Amazona parrots.</a> Wildl Soc Bull. 33, 756-760 (2005).</li><li>Vaughan, C., Nemeth, N., Marineros, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ecology+and+management+of+natural+and+artificial+Scarlet+Macaw+(Ara+macao)+nest+cavities+in+Costa+Rica.">Ecology and management of natural and artificial Scarlet Macaw (Ara macao) nest cavities in Costa Rica.</a> Ornitol Neotrap. 14, 381-396 (2003).</li><li>WCS Guatemala Program. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrated+interventions+to+conserve+Scarlet+Macaws+as+flagships+for+the+Maya+Biosphere+Reserve,+Guatemala.">Integrated interventions to conserve Scarlet Macaws as flagships for the Maya Biosphere Reserve, Guatemala.</a> 2009-2010 Report to BBC Wildlife Fund. , (2011).</li><li>Fish and Wildlife Service. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Puerto Rican Parrot (Amazona vittata) 5-Year Review: Summary and Evaluation. , (2008).</li><li>Berkunsky, I., DiazLuque, J. A., Kacoliris, F. P., Daniele, G., Milpacher, S., Gilardi, J. D., Martin, S., Eckles, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=10+Years:+Blue-throated+Macaw+Conservation.">10 Years: Blue-throated Macaw Conservation.</a> PsittaScene. 24, 3-5 (2012).</li><li>Synder, N., McGowen, P., Gilardi, J. D., Grajal, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Parrots: Status Survey and Conservation Action Plan 2000-2004. , (2000).</li><li>Delnicki, D. E., Bolen, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+Black-Bellied+Whistling+duck+nest+sites+by+other+species.">Use of Black-Bellied Whistling duck nest sites by other species.</a> SW Naturalist. 22, 275-277 (1977).</li><li>Twedt, D. J., Henne-Kerr, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Artificial+Cavities+Enhance+Breeding+Bird+Densities+in+Managed+Cottonwood+Forests.">Artificial Cavities Enhance Breeding Bird Densities in Managed Cottonwood Forests.</a> Wildl Soc Bull. 27, 680-687 (2001).</li><li>Carlile, N., Priddel, D., Zino, F., Natividad, C., Wingate, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+four+successful+recovery+programmes+for+threatened+sub-tropical+petrels.">A review of four successful recovery programmes for threatened sub-tropical petrels.</a> Mar Ornithol. 31, 185-192 (2003).</li><li>Oldroyd, B. P., Lawler, S. H., Crozier, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Do+feral+honey+bees+(Apis+mellifera)+and+regent+parrots+(Polytelis+anthopeplus)+compete+for+nest+sites.">Do feral honey bees (Apis mellifera) and regent parrots (Polytelis anthopeplus) compete for nest sites.</a> Aust J Ecol. 19, 444-450 (1994).</li><li>Downs, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Artificial+nest+boxes+and+wild+Cape+Parrots+Poicephalus+robustus:+persistence+pays+off.">Artificial nest boxes and wild Cape Parrots Poicephalus robustus: persistence pays off.</a> Ostrich. 76, 222-224 (2005).</li><li>Seeley, T. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+Nest+Cavity+Volume+by+the+Honey+Bee+(Apis+mellifera).">Measurement of Nest Cavity Volume by the Honey Bee (Apis mellifera).</a> Behav Ecol Sociobiol. 2, 201-227 (1977).</li><li>Schneider, S. S., DeGrandi-Hoffman, G., Smith, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+African+Honey+Bee:+Factors+contributing+to+a+successful+biological+invasion.">The African Honey Bee: Factors contributing to a successful biological invasion.</a> Annu Rev Entomol. 49, 351-376 (2004).</li><li>Seely, T. D., Visscher, P. K., Passino, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Group+decision+making+in+honey+bee+swarms.">Group decision making in honey bee swarms.</a> Behav Ecol Sociobiol. 94, 220-229 (2006).</li><li>Taylor, K. S., Waller, G. D., Crowder, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impairment+of+a+classical+conditioning+response+of+the+honey+bee+(Apis+mellifera+L.)+by+sublethal+doses+of+synthetic+pyrethroid+insecticides.">Impairment of a classical conditioning response of the honey bee (Apis mellifera L.) by sublethal doses of synthetic pyrethroid insecticides.</a> Apidologie. 18, 243-252 (1987).</li><li>Ingram, E. M., Augustin, J., Ellis, M. D., Siegfried, B. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluating+sub-lethal+effects+of+orchard-applied+pyrethroids+using+viedo-tracking+software+to+quantify+honey+bee+behaviors.">Evaluating sub-lethal effects of orchard-applied pyrethroids using viedo-tracking software to quantify honey bee behaviors.</a> Chemosphere. 135, 272-277 (2015).</li><li>Rieth, J. P., Levin, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+repellent+effect+of+two+pyrethroid+insecticides+on+the+honey+bee.">The repellent effect of two pyrethroid insecticides on the honey bee.</a> Physiol Entomol. 13, 213-218 (1988).</li><li>Efstathion, C. A., Bardunias, P. M., Boyd, J. D., William, K. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+push-pull+integrated+pest+management+scheme+for+preventing+use+of+parrot+nest+boxes+by+invasive+Africanized+honey+bees.">A push-pull integrated pest management scheme for preventing use of parrot nest boxes by invasive Africanized honey bees.</a> J Field Ornithol. 86, 65-72 (2015).</li><li>Seeley, T. D., Morse, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nest+site+selection+by+the+honey+bee,+Apis+mellifera.">Nest site selection by the honey bee, Apis mellifera.</a> Insect Sco. 25, 323-337 (1978).</li><li>Schmidt, J. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Attraction+of+reproductive+honey+bee+swarms+to+artificial+nests+by+nasonov+pheromone.">Attraction of reproductive honey bee swarms to artificial nests by nasonov pheromone.</a> J Chem Ecol. 20, 1053-1056 (1994).</li></ol>

The authors have nothing to disclose.

As a highly successful invasive insect17, AHB constitute a major competitor for cavity-nesting birds throughout their range. The protocol presented here is designed to reduce the colonization rates of bird nest boxes by AHB. The main components of this design include 1) the use of a repellant and bird-safe insecticide, permethrin; 2) The provision of an alternate nesting site for bees; and 3) the use of an attractant pheromone inside trap boxes. Bees usually make the &#34;right&#34; decision when choosing a ne...

Since the accidental introduction of more than two dozen African honey bee queens into Sao Paulo, Brazil in 19571, these insects have spread throughout South and Central America and up into the southern United States, usurping and replacing the less well adapted European honey bees (EHB) in the feral population. These hybrid Africanized honey bees (AHB) have traits that put them in conflict with both humans and wildlife. They exhibit highly defensive behavior, reproduce very fast, and are less selective about where they make a home1,2.
Although nest boxes have long been used by ornithologists to enhance breeding by cavity-nesting birds3,4, their use has been shown to be especially effective for enhancing the breeding of threatened and endangered parrots in the tropics5,6. Unfortunately, Africanized honey bees often choose these same nest boxes&#160;&#8211; sometimes even killing the occupants. Additionally, these AHB pose a significant threat to biologists when they are conducting nest studies. In areas of South America where AHB have been established, several parrot conservation projects have seen their artificial nest boxes occupied by AHB7-10. Competition for nest boxes with honey bees is widespread and is not confined to parrots. Many other bird species both in the New and Old World are affected11-15. Without efforts to mitigate the threat of these invasive AHB, recent successes in parrot conservation could be lost.
Current practices for managing competition from bees includes the use of repeated, high doses of permethrin7, manually removing swarms, closing nest entrances immediately after breeding season8, or the application of chemicals to the outsides of nest boxes15. The labor involved and the danger to biologists render these strategies less than optimal. Development of a prevention protocol is warranted.
Honey bee colonies multiply via a process known as swarming, when the old queen leaves the colony with up to three fourths of the worker bees16. Africanized honey bees&#39; high reproductive rate allows them to swarm more frequently (four to eight times per year) than EHB (who swarm once or twice per year)17. Moreover, AHB may swarm throughout the year rather than during a particular season like EHB. Consequently, AHB can quickly occupy most of the suitable nesting sites in a given area.
Honey bee house hunting behavior can be utilized to influence their choice of nest site. A small fraction of bees in a swarm fly off to locate and inspect new potential nest sites. Several potential nest sites are scouted for simultaneously. These scout bees return to the swarm and perform dances to indicate the location of the site. The stronger the dancing is for a site, the faster the number of scout bees builds up at that site18. Therefore, bees dance stronger for better quality sites, compared to sites that are mediocre or poor. Once a quorum threshold is reached, the swarm takes off and flies to the new chosen home site18.
Sub-lethal contact doses of the insecticide permethrin hamper a scout bee&#39;s ability to orient and reduce its activity level19,20. An application of permethrin inside nest boxes at first should be repellant to scout bees21, and overtime as the chemical degrades, scout bees exposed to permethrin during their assessment of the box, should be rendered less able to perform strong recruitment dances, decreasing the amount of scout bees recruited to the site. A more suitable (ideal cavity volume, small entrance hole located at the bottom, and a pheromone attractant) alternate nest site is provided in the form of a trap box for the bees. This excellent trap site should reach a quorum faster than the insecticide treated bird box leading the swarm to choose the trap box over the bird box.
As an example, we describe an experiment to reduce colonization of Barn Owl artificial nest boxes by AHB in South Florida sugarcane fields. Our objective was to test the effectiveness of using a push-pull method that will &#34;push&#34; bees away from owl nest boxes treated with an insecticide and &#34;pull&#34; them toward pheromone baited swarm traps.

<table border="0" cellpadding="0" cellspacing="0" width="652">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Citral (ACROS Organics)</td>
			<td style="width:117px;">Fisher Scientific</td>
			<td style="width:119px;">AC11044-1000</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Geraniol (ACROS Organics)</td>
			<td style="width:117px;">Fisher Scientific</td>
			<td>AC41090-1000</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">PermaCap CS</td>
			<td style="width:117px;">BASF</td>
			<td style="width:119px;"></td>
			<td>controlled release permethrin</td>
		</tr>
	</tbody>
</table>

a push-pull protocol to reduce colonization of bird nest boxes by honey bees

Introduction of the invasive Africanized honey bee (AHB) into the Neotropics is a serious problem for many cavity nesting birds, specifically parrots. These bees select cavities that are suitable nest sites for birds, resulting in competition. The difficulty of removing bees and their defensive behavior makes a prevention protocol necessary. Here, we describe a push-pull integrated pest management protocol to deter bees from inhabiting bird boxes by applying a bird safe insecticide, permethrin, to repel bees from nest boxes, while simultaneously attracting them to pheromone-baited swarm traps. Shown here is an example experiment using Barn Owl nest boxes. This protocol successfully reduced colonization of Barn Owl nest boxes by Africanized honey bees. This protocol is flexible, allowing adjustments to accommodate a wide range of bird species and habitats. This protocol could benefit conservation efforts where AHB are located.

A previous small pilot push-pull study conducted in Brazil showed swarm traps were effective and reduced bee occupation of parrot nest boxes22. To determine if the push-pull method can be effective at a larger scale to reduce colonization of bird nest boxes by AHB, we conducted a field study in sugarcane fields in southern Florida, USA. Sixty Barn Owl nest boxes were installed and each was paired with a swarm trap located 15 m away. Every swarm trap received a pheromone lure. O...

Watch this Scientific Journal Video about A Push-pull Protocol to Reduce Colonization of Bird Nest Boxes by Honey Bees at JoVE.com

A Push-pull Protocol to Reduce Colonization of Bird Nest Boxes by Honey Bees

Preventing colonization of bird nest boxes by invasive Africanized honey bees is important for conservation efforts of nest-site limited birds. We provide an integrated pest management approach to &#34;push&#34; bees away from nest boxes with a repellant insecticide, permethrin, and &#34;pull&#34; them toward pheromone baited swarm traps.

Introduction of the invasive Africanized honey bee (AHB) into the Neotropics is a serious problem for many cavity nesting birds, specifically parrots. ...

The overall goal of this procedure is to prevent honey bee colonization of bird nest boxes by using insect repellent in the bird nest boxes and a pheromone lure in the swarm trap boxes. This method can help solve the conservation issue of Africanized honey bee hybrids competing for cavities with cavity-nesting birds. The main advantages of this technique are that it's safe for the birds, deters honey bees without the need to kill entire colonies or swarms and deters blood-feeding ectoparasites.We first had the idea for this method when we had to deal with an Africanized honey bee colony in a screech owl nest box here in South Florida. Visual demonstration of this method is critical as the implementation of this procedure varies according to the environmental and bee colony circumstances. Before placing the swarm trap box, add one milliliter of a two to one mixture of citral and geraniol into a 1.5 milliliter snap top polypropylene microcentrifugation tube and wrap the tube in aluminum foil to protect the lure from light.Then store the lure in a dark place at room temperature. To install the swarm trap on a post, fix a nine by nine centimeter pressure-treated 2.4 meter long post into the ground, 10 to 15 meters from where the bird nest box is located. Next, use at least two screws to secure a 1.3 centimeter thick 20 by 20 centimeter pressure-treated wooden hanging board in the middle of one side of the trap box.With the top of the board, set 10 centimeters above the top of the box. Then place a screw approximately five centimeters below where the top of the hanging board will rest and attach the hanging board to the post such that the trap box is 1.5 to three meters above the ground to allow the safe and easy removal of the box after colonization. To install the swarm trap box on a tree, first drill a five millimeter hole in the upper back corners of each side of the box and thread a wire through the holes.Next, select a tree approximately 10 to 15 meters away from where a bird nest box is located and secure the wire around the tree 1.5 to three meters off the ground. Then confirm that the box is level and will not tip over. When the swarm trap box has been secured, use a pushpin to puncture the cap of the tube of lure and place the lure at the bottom of the trap box near the entrance.Then use a 3.7 liter hand pump sprayer to coat the entire inside of the bird nest box with a freshly-prepared permethrin for poultry or caged birds until the liquid runs off the sides of the box. If the swarm is to be utilized, allow the bees to fully colonize the trap box for about two weeks. Then at night when all of the bees are inside, don the appropriate protective equipment and use a small towel to plug the trap box entrance.The next morning collect the box, move it to a new location at least four kilometers away and remove the towel. If destruction of the bee colony is warranted, as soon as the colony is discovered, plug the entrance hole with a small towel and remove the box. Seal the trap box completely in a plastic bag and leave the colony in the full sun.After several days remove the dead bees and comb. If the use of an insecticide is preferred, as soon as the colony is discovered, spray the inside of the trap box with an insecticide spray and plug the entrance with a small towel. After 24 hours, remove the dead bees and comb.Of the 77 swarms caught in this representative experiment, 75 were in swarm traps. Only two owl nest boxes were colonized, both of which were untreated. Significantly more bees chose swarm traps over untreated or treated owl boxes and several swarm trap boxes were colonized by bees more than once.Once mastered, this technique can be completed in under one hour if it is performed properly. While attempting this procedure it's important to remember to be safe. Always place the swarm trap boxes where they will be easy to remove and relocate them at night when the bees will not be active.This procedure can be modified to accommodate a wide range of bird species and habitats. After its development, this technique paved the way for researchers in the field of conservation to explore tools for managing the bee swarm colonization of bird nest boxes in recovery programs of endangered parrots. After watching this video you should have a good understanding of how to set up bee swarm trap boxes and to safely remove and relocate or destroy the bees that have colonized the trap box.Don't forget that working with Africanized honey bees can be extremely hazardous and that precautions such as wearing bee suits and gloves should always be taken while performing this procedure.

a-push-pull-protocol-to-reduce-colonization-bird-nest-boxes-honey

Research

JoVE Journal

Environment

8.5K Görüntüleme.  University of Florida. The overall goal of this procedure is to prevent honey bee colonization of bird nest boxes by using insect repellent in the bird nest boxes and a pheromone lure in the swarm trap boxes. This method can help solve the conservation issue of Africanized honey bee hybrids competing for cavities with cavity-nesting birds. The main advantages of this technique are that it's safe for the birds, deters honey bees without the need to kill entire colonies or swarms and deters blood-feeding ectoparasite...