Managing Varroa Mites in Colonies of Honey Bees By Jeff Harris, Mississippi State Extension In recent years honey bee health has become a primary focus of researchers in response to several episodes in which commercial colonies were lost at unusually high rates in the U.S. and Canada. Although not fully understood, high colony mortality stemmed from multiple factors that included the parasitic mite Varroa destructor Anderson & Trueman, viruses vectored to bees by varroa mites, pesticide exposure, residues of agrochemicals in hives, and poor nutrition. Varroa mites and the viruses they vector are currently viewed as the primary killers of honey bees worldwide. The acaricides used to control varroa mites are an additional threat when sequestered as chemical residues in combs. Chemicals placed in colonies contaminate hive products (e.g. beeswax) and may cause sub- lethal effects on individual bees that reduce overall colony health. Use of miticides also results in mite populations with genetic resistance to these chemicals. For these reasons, use of chemicals to control varroa mites should be minimized. This goal is best achieved by implementation of integrated pest management (IPM) strategies that slow the growth of mite populations and reduce the need for miticides to control the mites. This pamphlet provides an overview of the biology of varroa mites and IPM approaches that can be used to manage varroa mite populations. Basic Biology of the Varroa Mite The varroa mite is the most serious pest of the Western honey bee (Apis mellifera L.) throughout the world. This mite was first described on the Eastern honey bee (Apis cerana F.) from Java in 1904. The mite was originally called Varroa jacobsoni Oudemans, but research published in 2000 indicated that there existed more than one species of varroa mites. It turns out that V. jacobsoni usually does not kill or damage colonies of the Western honey bee, and V. destructor is the species causing most harm to bees. The first reports of varroa mites attacking Western honey bee were in the early 1960s in Asia. The varroa mite has since become established on every continent except Australia, although the mite will likely arrive there very soon. Varroa mites spread between continents via commercial transport of bees by beekeepers, and by movement of swarms that fly long distances or hitch rides on ships, airplanes and trucks. Varroa mites move within infected regions by the same transport mechanisms. Within apiaries, varroa mites move with drifting bees, or they hitch rides on robber bees that attack hives weakened by mite parasitism. Figure 1 – Adult female varroa mites live on adult bees when not reproducing in the capped brood cells (photo by Stephen Ausmus, USDA, ARS). Adult female varroa mites are oval and flat, about 1.1 mm long and 1.5 mm wide. They are tan to reddish-brown in color and can be seen easily with the unaided eye (Fig 1). Male mites are considerably smaller and are lightly tan in color. Male mites are not usually seen outside brood cells. Varroa mites reproduce within brood cells where they feed upon the developing bee pupae to obtain nourishment for egg production (Fig 2). Mite offspring also feed on the host pupa to obtain nourishment for growth and development. When colonies of bees are raising drone brood, varroa mites prefer to invade drone brood about 9-times more than worker brood. This is significant because the development time of drones is a few days longer than workers (24 and 21 days, respectively), and the mites can produce more adult daughters when in drone brood, and mite populations can grow faster than when a colony only has worker brood. Adult bees serve as intermediate hosts when there is no brood available. Varroa mites attach to adult bees between the abdominal segments or between body regions (head-thorax-abdomen), making them difficult to detect. These are also places from which they can easily feed on the hemolymph or blood of the adult bee. Figure 2 - Life cycle of the Varroa Mite. Adult female mites live on adult bees for an average of 1 week between attempts at reproduction in brood cells. Each mite will produce 3-4 daughters and 1 son when reproducing in a worker brood cell. Usually, only the mother mite and 1-2 adult daughters leave the brood cell with the emerging bee. Each adult female mite will attempt 3-4 reproductive bouts in her life. Adult female mites invade brood cells that contain 5-day old worker larvae, and the adult bees cap these brood cells shortly afterward. Each mite lays up to 4 or 5 eggs at approximately 30- hour intervals. The first egg usually develops into a male, and the later ones into females. The development of offspring proceeds from egg to six-legged larva (within the egg envelope), to eight-legged protonymph, to deutonymph, and finally to a sexually mature adult mite in 5.5 days (female) to 6.5 days (male) (Fig. 3). The larvae and protonymphs of male and female mites look identical, but the deutonymphs and adults are easily differentiated. The females are oval in shape, while the males are more triangular in shape. Mites shed their cuticles when molting into the next nymphal stage. However, only the shed cuticles from the last molt (deutonymph into adult mite) are easily seen in brood cells. Young adult mites mate in the capped cells. Adult males and all immature mites die after the host bee emerges from the brood cell (when nest bees clean the cells). Young adult daughters and the mother mite exit the brood cell with the emerging host, and they eventually transfer to other passing bees. A mite enters another brood cell in ca. 7 days during periods when brood is available. However, varroa mites may survive over >100 days when no brood is available, such as during the winter months. Figure 3 – Developmental stages of varroa mites. Varroa mites develop through a larval stage that has six legs, and after “hatching” from the egg, each mite progresses through two nymphal stages. Adult bees growing from a mite-infested brood cell suffer from loss of blood and are often underweight. Their lifespans may be greatly reduced, and the most severe damage occurs when mites vector harmful viruses to the bees. Although many viruses can be vectored, it seems that a particularly virulent form of the honey bee Deformed Wing Virus (DWV) kills most colonies. The degree of damage caused to be pupae from direct feeding by the mites depends on the number of mites parasitizing each bee larva. One or two invading mites will produce enough offspring to decrease the vitality of the emerging bee. Higher numbers of varroa mites per cell result in malformations like shortened abdomens, misshapen wings, and deformed legs or even in the death of the pupa. Normal colony functions become disrupted when young bees that normally progress through a series of age-related tasks are lost prematurely from the bee population. Eventually, the mite population increases to economically damaging levels, and a colony of bees weakens and begins to exhibit symptoms of a condition known as parasitic mite syndrome (PMS). The basic symptoms are a very weak colony with brood combs having a poor capped brood pattern and sick larvae showing a yellowing or browning color caused by viral diseases (Fig 4). Usually a colony exhibiting PMS is lost and cannot be easily saved. Figure 4 – Parasitic Mite Syndrome (PMS) from a highly mite-infested colony. The open brood cells show sick and yellowing larvae suffering from viral diseases. Loss of these individuals results in a capped brood pattern showing many empty cells (photo by Audrey Sheridan, Mississippi State University). Integrated pest management (IPM) and Varroa Mites IPM is a method of dealing with pests and parasites that differs from the application of chemical treatments on a regular schedule. It is based on the realization that one cannot chemically eradicate pests or parasites but must continually manage their populations. IPM (Fig 5) involves mixing different tactics (genetic/cultural methods, mechanical/physical methods and chemical treatments) and knowing critical times in the life cycles of the target organisms that make them vulnerable to control methods. Chemicals are only applied when pest populations are above an action or economic threshold, rather than on a timed schedule. The reduced use of chemicals to control varroa mites limits possible contamination of hive products and delays the development of resistance to the chemical by the mites. It also reduces the exposure of the bees to potentially harmful chemicals. This is especially significant because most of the miticides used to kill varroa mites are known to have some negative health consequences for honey bees. Sampling for Varroa Mites A key component of IPM is that decisions to use chemicals for controlling pests are based on sampling (monitoring and identification) the pest population and only treating when a critical threshold has been reached. Generally, the action threshold reflects a population of mites that if not treated could quickly grow to damaging levels that cause economic injury to your hive. The decision on whether to apply or not to apply a chemical is based on thresholds derived from sampling the pest population. In beekeeping, this is not as easy as in row crops, but is still possible. Figure 5 – The IPM Pyramid. The goal of integrated pest management is to combine several activities at the base of the pyramid to prevent or slow pest populations from reaching damaging levels that require intervention with a chemical treatment. Monitoring or sampling is used to make treatment decisions. If a chemical treatment is warranted, the choice for intervention should be of the least toxic or harmful chemical available. The economic threshold for varroa mite populations in colonies with 25,000 – 34,000 bees is somewhere in the range 3,172 – 4,261 total mites in August for the southern U.S. This range corresponds to 15-38 mites found on a sample of 300 adult bees, which is a mite load of 5-12%. Most beekeepers prefer to use the 5% end of the range as the action threshold. Migratory beekeepers may need to use a lower threshold because there is a lot of movement of mites between holding yards and among apiaries from different commercial beekeepers at pollination sites. Additionally, research suggests that some viruses vectored by varroa mites are becoming more virulent through time. More research is needed to determine the best minimal thresholds for a variety of beekeeping practices. Until new thresholds are established, it might be prudent to intervene when the total mite load in a colony is 2,000 – 2,500 mites, which is a population only weeks from a currently accepted economic threshold (Table 1). The mite population is distributed among the adult bees and the capped brood within a colony. Although there is considerable variation, typically one third of the mite population resides on adult bees, and two thirds of the mites are in capped brood. This distribution assumes that brood production is normal and has not been interrupted by loss of a queen. This assumed distribution is probably less accurate when drone production is highest in early spring, but it provides a reasonable basis for interpreting the different sampling methods to be described below. Table 1 was generated by assuming the economic or action threshold is the midpoint of 3,700 mites for the range of 3,172 – 4,261 mites. The mite load on adult bees was calculated by assuming one-third of all mites were on bees, and the bee population was 24,000 – 34,000 adult bees. The time to reach 3,700 mites was estimated using exponential growth rates measured for mite populations in a typical spring-summer season (not during hot and dry periods associated with drought) in the southern U.S. Table 1 - Miticide Treatment Threshold for Varroa Mites Total Mite Mites on Time of Beekeeper Action2 Year Population Adult Bees1 Wait; 9—12 weeks for mite population to 500 mites 0.5 – 0.7% reach threshold Wait; 6—8 weeks for mite population to summer 1,000 mites 1.0 – 1.4% reach threshold Treat with miticide; 2—3 weeks for mite 2,000 mites 2.0 – 2.8% population to reach threshold Treat with miticide; 1 – 2 weeks for mite autumn  2,500 mites 2.4 – 3.3% population to reach threshold Treat with miticide immediately; at or above anytime  3,700 mites 3.6 – 5.1% economic threshold 1Range given for colony population equal to 24,000 – 34,000 adult bees. 2Range based on mite population growth rates during typical and non-drought seasons in the southern U.S. A. Sampling Mites on Adult Bees The most convenient methods of sampling varroa mites involve estimating the prevalence of mites on the adult bees. The prevalence is often described as a percentage. For example, a 5% mite load means that there were 5 adult mites counted for every 100 bees in the sample. Most beekeepers prefer not to kill bees during sampling, but people with many colonies might find it more efficient to collect samples from many colonies at once and measure the mite loads at a more convenient time later. The most commonly used non-destructive sampling method is the powdered sugar shake. The destructive method involves washing samples of dead bees with alcohol or a soapy water mixture. 1. Sugar Shake Method: The powdered sugar roll or sugar shake provides a means for estimating the mite load without killing the adult bees that are sampled. The method hinges on the ability of dusts with small particle size to dislodge the adult mites from the bodies of the adult bees. The small particles of 10× confectioners’ sugar create a barrier between the bodies of bees and the specially modified feet of the mites, which are like small suction cups. The first step is to make a jar for sampling adult bees. A wide-mouthed quart or pint sized canning jar works nicely. The trick is to replace the solid dome of the canning jar with a circular piece of 8×8 wire mesh cut to fit the jar. The mesh is held in place with the ring normally used to secure the canning dome lid. When bees are rolled and coated in the sugar, the feet of the mites eventually contact the powder and they lose their grip. The sugar can then be poured onto a sheet of wax paper, and the falling mites can be counted. If it is windy, the sugar can be sprinkled onto a heavy duty white paper plate that resists being blown away. To sample a colony, it is best to locate the queen and make sure she does not accidentally end up in the sampled bees. Varroa mites reside on younger bees within the broodnest. Choose two combs with capped and/or uncapped brood from the brood chamber and gently shake the adhering bees from the two combs into an empty box (e.g. a cardboard nuc box works well). Mix the bees and then take 400 bees (½ cup) as a sample and place them into the sampling jar. Secure the mesh wire with the canning ring and add 2 tablespoons of powdered sugar through the mesh onto the bees. Mix the bees and sugar to thoroughly coat all of the bees. Wait about 2-3 minutes before turning the jar on end and shake the sugar from the jar onto the wax paper or paper plate. Continue shaking the bees for a minute or so to dislodge all of the mites. A second cycle of adding sugar and shaking can be used to verify that all mites have been dislodged from the bees. The sugar-coated bees can be returned to their colony after sampling, and they will be cleaned by nestmates. The mites will appear as small brown oval flecks in the powdered sugar. Sometimes movement of their legs makes them easier to see. If you have trouble seeing the mites, a small amount of water can be added to dissolve the sugar and reveal the mites. To estimate the mite load, count the total number of mites dropped from the sample and divide by the total number of bees that were sampled and multiply by 100. For example, (20 mites divided by 400 bees) times 100 equals 5%. 2. Alcohol wash: The alcohol wash is similar to the sugar shake except that the adult bees are killed prior to measuring the mite load. Bees are taken from two combs of capped brood as in the previous section, and 400 worker bees are put into a jar or into a Ziploc bag, which is then sealed. Jars or bags should be labeled to identify each colony being sampled, and samples can be stored on ice during transport from an apiary. Samples in Ziploc bags can be frozen if washing will be done at a later date. When you are ready to wash frozen samples, transfer the bees from the Ziploc bags into canning jars and add 70% ethanol to completely cover the bees. Be sure to examine the Ziploc bags for any mites and transfer them into the jars with the bees. If bee samples are collected in canning jars while in the field, 70% ethanol can be added to them immediately, especially if they are to be washed on the following day or so. The level of alcohol put into each jar should completely cover the adult bees. Secure the solid dome lid onto each jar with the ring. Washing works best if the bees have soaked in the alcohol for at least a day. The bees should be shaken vigorously for several minutes before being poured through a filter made of 8×8 wire mesh placed over a catch reservoir. The bees will be retained by the mesh, while the ethanol and mites pass into the reservoir (Fig 6). If the reservoir is white, the mites show well for counting. After counting the first wash of mites, decant the ethanol from the sample and discard the mites. Wash the sample of bees again. Several washes may be needed to dislodge all mites. The mite load is found as in the previous section by dividing the total number of mites by the total number of bees that were sampled and multiplying by 100%. Figure 6 – Alcohol wash of varroa mites from a sample of adult worker bees. The bees are retained by a mesh filter while the 70% ethanol and mites pass to the reservoir. The white color of the reservoir allows easy counting of the mites. If you do not have access to ethanol, a soapy water solution can be used to wash the mites from the bees. Add 1-2 ml (15-30 drops) liquid dish detergent to 500 ml (ca. 1 pint) of water to make the washing solution. Agitate the bees in the detergent solution for several minutes before filtering. Longer agitation periods by placing the jars of bees onto mechanical shakers for several hours will increase the efficiency of this method. B. Using Mite Drop to the Floor as Indicator of Mite Population The increased popularity of screen floors in hives as a non-chemical control method for varroa mites (see below) offers a fairly passive and relatively non-disruptive method for sampling colonies. The method is based on the phenomenon that the number of varroa mites falling to the floor correlates to the mite population within the colony. Specifically, the degree of “mite drop” reflects the total mite population – mites in brood cells and on adult bees. Presumably, adult female mites fall from worker bees regularly, and dead adult mites are removed from brood cells and dropped to the floor by nest cleaning bees. The sugar shake or alcohol wash only estimates the mite load on the adult bees. The basic procedure is to insert a sticky cardboard covered with adhesive beneath the screened floor of the hive. The 8×8 screen mesh allows mites to fall through to the adhesive on the sticky board but prevents the bees from contacting the sampling surface. The adhesive may be a thin layer of non-stick cooking spray or petroleum jelly applied to the board. Many commercial screened bottom boards are made with a groove to which a sticky board can be easily inserted from behind the hive. The commercial sticky boards usually come with a sampling grid to make counting the mites easier. Sticky boards can also be made from white cardboard and cut to fit the typical bottom board. The sticky board is left in place for 3 days, and the number of dead mites is counted. The daily “mite drop” is estimated by dividing the total mites by the number of days for the sampling period. The economic thresholds estimated for using this technique vary wildly from region to region. However, a reasonably conservative threshold for late summer in Mississippi would be a daily mite drop >40-50 mites per day. C. Making Apiary-Level Treatment Decisions Practical sampling plans have been developed that beekeepers can use to make treatment decisions for whole apiaries based on samples from a few colonies (Lee et al. 2010). Commercial beekeepers needing to make apiary-level treatment decisions do not have the time or resources to sample every colony they own, which could be hundreds to thousands of colonies. Although sampling schemes were developed to make apiary-level treatment decisions, a beekeeper cannot make a decision about treating all of his apiaries based on the sampling decision made for one apiary. This is because the mite loads can vary significantly between apiaries within a single commercial operation. The basic procedure for apiary-level decisions involves averaging the adult bee mite loads taken from a sub-set of colonies within the apiary – each colony is sampled by an alcohol wash of 300- 400 bees (as above). The average mite density of these samples will reflect the average mite load of the entire apiary. If the average mite load exceeds the action threshold, the entire apiary is treated with chemicals to control varroa mites. The number of colonies sampled in an apiary hinges on the number of colonies within the apiary. The colonies sampled should be chosen at random and should span the entire geographical range of the apiary (not just concentrated in one area). For example, apiary-level treatment decisions can be made for apiaries with > 20 colonies by sampling no more than 8 colonies with a reasonable level of precision. This means that a beekeeper samples 8 different colonies in an apiary of 25 colonies, and he only samples 8 colonies in another apiary of 60 colonies. The researchers that developed the schemes did recommend fewer samples for smaller apiaries (Table 2). Table 2 – Number of Colonies to Sample for Apiary-Level Treatment Decisions No. Colonies No. in Apiary Sampled >20 8 20 6 10 5 4 3 Non-chemical Methods for Slowing Growth of Mite Populations Various non-chemical methods can be used to reduce the growth of mite populations in colonies of honey bees. For example, immigration of mites between apiaries can be limited if the distance between apiaries is increased from 1-2 miles to several miles. Similarly, the distance between colonies within an apiary can be increased from a few feet to twenty feet or more to reduce the drift of varroa mites among colonies. Placing colonies in irregular groups rather than straight rows will also reduce the drift of bees and mites between colonies. Beekeeping practices may need to be changed to avoid enhancing mite populations. For example, beekeepers commonly combine weakened colonies with stronger ones to save the hive equipment (e.g. to keep combs from being damaged by larvae of the Small Hive Beetle). This practice may actually transfer varroa mites and viral diseases to the stronger colony. Each beekeeper should evaluate his or her beekeeping routines to decide if certain procedures need modification to avoid doing things that inadvertently worsen the varroa mite problem within the apiary. Other strategies to combat varroa mites include the use of dusts such as powdered sugar to remove mites from all of adult bees in a colony, and purposely creating a break in brood production by caging queen honey bees. Although these methods may be useful under some conditions, they are relatively cumbersome to implement and effectiveness may be limited. The following non-chemical methods are either popularly used by beekeepers, or have been found to control varroa mite populations by multiple researchers. A. Trapping Mites in Drone Brood These methods take advantage of the natural preference that varroa mites have for drone brood over worker brood. They prefer drone brood about 9-times more than worker brood, and because of the longer development time for drones (24 days) versus workers (21 days), each