How effective propolis is on preventing bee viruses.

Brianna Breiland, Amy Puckett, Paige Salmon, Nicole Wienke1University of Wisconsin-River Falls, WI

Introduction

The honey bee, Apis mellifera, is an insect that makes honey by going around to flowering plants and trees; they then collect the nectar from the flowering plants and bring it back to the hive[1]. They are transported all around the United States to pollinate crops and plants. One third of the world’s agricultural production is dependent on honey bees. If a honey bee finds a good source of food it does a series of movements or a dance to show exactly where the food can be found. The waggle dance tells the bees what direction and how far the food source is from the hive.  The bees produce a product called propolis with in the hive. Propolis is made from bees collecting the sticky substance that trees produce to protect their buds from fungus and infection.  After the bee brings that sticky substance back to the hive it is used to seal cracks, smooth rough wood, and encase foreign objects with in the hive[3]. Our hypothesis is that the more propolis formed in the hive the less virus will be detected in the hive. In theory, hive containing a propolis envelope will have minimal virus infection when compared to the other 2 hives. Recently there has been an increase in what is believed to be a virus called Colony Collapse Disorder (CCD). CCD is a phenomenon in which worker bees from a beehive or honey bee colony abruptly disappear. This disorder was noticed in the United States around 2006 and is a serious problem threatening the health of honey bees and the economic stability of commercial beekeeping and pollination operations in the United States[2]. Despite a number of claims in the general and scientific media, a cause or causes of CCD have not been identified by researchers. Due to CCD more than 30 percent of the honey bee colonies have died throughout the United States. There has been a world-wide decline in the population of honey bees and other native pollinators. There is not a single known cause to CCD but there are many factors that contribute to it. Those include poor nutrition, pesticides, and Israeli Acute Paralysis Virus (IAPV)[4]. Honey bees suffer from multiple viruses but the three which are research focused on are Black Queen Cell Virus (BQCV), Deformed Wing Virus (DWV), and Israeli Acute Paralysis Virus (IAPV), all three of which are an RNA virus affecting the honey bee, Apis mellifera. BQCV causes the queen larva cells to turn dark brown or black and die. The virus is thought to be associated with Nosema, a spore forming unicellular parasite. Nosema significantly affects the bees and causes the colony to fail to thrive as a result. DWV was isolated from symptomatic bees, located in Japan, in the early 1980’s, causes wing and abdominal deformities. These deformities are often found on adult honey bees infested with varroa mites. Symptoms include damaged appendages, stubby, useless wings, shorted and rounded abdomens, discoloring and paralysis of the legs and wings. Symptomatic bees have a severely-reduced life span and are typically thrown out of the hive to reduce the spread of the virus. IAPV first arrived in the United States in 2006 due to the transport of infected honey bees from Austrailia in 2005. IAPV is closely associated with CCD from the amount of IAPV genetic material in bees suffering from Colony Collapse disorder.

ConclusionReferences

1. Honey bee, Apis mellifera. Available from: http://www.fcps.edu/islandcreekes/ecology/honey_bee.htm

2. USDA: CCD. Available from: http://www.ars.usda.gov/news/docs.htm?docid=15572 3. Basic Beekeeping. Available from: http://

basicbeekeeping.blogspot.com/2011/12/lesson-113-sticky-subject-of-propolis.html

4. Saving the honey bee, Scientific American, April 2009, page 40-47

5. Biology 9th edition, McGraw Hill,2011. Avaliable from: http://www.bio.davidson.edu/genomics/method/cDNAproduction

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Figure 1. The picture on the left illustrates the propolis formed on the sides of the hive by the honey bees in their colony. The picture on the right shows the multiple features of a honey bee.

Materials and MethodsDuring September 2012, honey bees were collected from three apiaries located in Pierce and St. Croix counties. The various apiaries were either a control hive containing no propolis, a propolis trap, and a propolis

envelope. Place 5 bees in a sterile petri plate. Remove abdomen of one bee and transfer the abdomen into a Kaypak bag in an ice bucket. Sterilize the scalpel and forceps 3 times before removing the next abdomen. Cut abdomen and repeat. Label 4 micro-centrifuge tubes with the bee number, initials and date. In addition add the bee number to the lids. On three of the tubes write RNA and on the fourth write “Lysate.” Add 0.5 ml Lysis Buffer per bee to the bag dispensing the buffer inside the bag to avoid wetting the top of the bag. Heat seal the bag, pushing out as much air as possible without getting the top of the bag wet. Using an empty 50 ml tube as a rolling pin, smash the bee abdomens in the bag thoroughly, mashing them for about 30 sec until they are completely homogenized except for the exoskeleton. Massage the liquid in the bad to mix fully, then draw off 620 µl of liquid and transfer it to one of your labeled micro-centrifuge tubes, avoiding bubbles.

Colony Numbers BQCV DWV IAPV6X - - -12.92

20 Bee Pool -12.32 -11.38 -18.787X -9.51 -8.94 -20.38

20 Bee Pool -9.44 -11.76 -15.4313

X -10.14 -14.8620 Bee Pool -10.66 -4.48 -17.02

14X -12.19 -3.55 -18.61

20 Bee Pool -10.43 3.73 -17.5317

X -12.69 -4.8925 -19.1520 Bee Pool -11.3 2.71 -18.91

18X - -10.27 -9.53

20 Bee Pool No Ct No Ct -10.8222

X - -7.035 -14.8420 Bee Pool No Ct -7.66 -12.64

23X -10.02 -4.213 16.206

20 Bee Pool No Ct -3.196 -17.1826

X -11.426 -11.443 -17.7220 Bee Pool -9.61 -8.656 -16.97

28X - -11.54 -13.56

20 Bee Pool -10.67 12.41 -15.46630

X - -9.265 -17.4720 Bee Pool -10.55 -2.906 15.6933

33X - -13.125 -19.3

20 Bee Pool -10.525 -15.506 -15.7136

X -4.5325 -14.88520 Bee Pool -10.27 1.336 -14.4367