Honey Bee Diseases Christopher Cripps, DVM Betterbee Grennwich, NY

Honey Bee Diseases Christopher Cripps, DVM Betterbee Grennwich, NY Honey bee disease etiologies Bacterial o American Foulbrood o European Foulbrood Fungal o Chalkbrood o Nosema Parasitic o Varroa mites o Tracheal mites o Small Hive beetles o Tropilaelaps Viral o Over 20 different viruses Toxicities o Pesticides Idiopathic o Colony Collapse Disorder Environmental o Starvation o Excess Humidity Varroa destructor Foreign Animal Disease in USA prior to 1987 Now very common across USA If bees sick, this is the first differential Reproduce well with honeybees Carry diseases Complex control methods, no silver bullet Bee keepers... Bee havers Bee hadders o Life b.m. o Tracheal mites 1984 o Varroa mites 1987 Varroa destructor life cycle Mites on bees = Phoretic mites Female leaves bee, enters brood about to be capped Lays eggs starting 60 hours after capping Varroa life cycle First egg = male (haploid) Lays egg every 30 hours Rest are female Feed on pupa Mate as adults in cell Emerge with adult bee Drone brood preferred o ~15 days available o Worker ~ 12 days o Extra 2 mites per time in brood 450

Population dynamics Stop adding bees Keep adding mites More bees removed o worked to death Higher mites per bee More disease spread because more chance an infectious mite bites a bee. Varroa What problems caused? Puncture the exoskeleton of the bee Wound caused, and immuno-suppressors injected. Feed on fat bodies Dirty mouth parts? Contaminated saliva? Opportunistic invaders? Now over 20 viruses recognized in bees, most are spread by Varroa mites Immuno-suppression Viral Loads o More mites, more spread o Infective Dose varies by route of administration o Enhancement by mite of viral infectivity? o Types of viruses seen DWV, BQC, IAPV, CPV Decreased lifespan o More disease, more accidents, less exercise tolerance (oxygen carrying capacity) Birth Defects? Varroa treatments Originally: burn hives until they were eradicated Then: Treat with harsh chemicals 1x or 2x per year Now: o Monitor level of mites. o Treat if level too high. Soft chemicals or hard chemicals. o Continue other control methods. o Recheck mite levels Varroa mite monitors Visual bad! Drone brood exam Sugar Shake Ether Roll/alcohol wash Sticky Boards Varroa control - Chemicals Integrated Pest Management Chemicals often seen as last resort. Chemicals often used to treat everything whether needed or not. Concerns: o Kill mites, not bees Some side effects bees, brood, queens, sperm o Honey is a food product o Wax is considered a chemical sponge Studies show wax has a lot of residues, many are beekeeper applied miticides. Varroa mite control varroa mite control Natural Methods Drone Comb Removal 451

o Remove breeding mites Breeding resistant bees o Russian bees, Varroa Sensitive Hygiene, ankle-biters Swarming Splitting o More time without brood where mites hide and reproduce. Phoretic mites exposed to bees. Common clinical presentations My bees were very strong, now they are all gone I was new this year, so I did not have to worry about mites. Why are my bees all dead? I have honey in the hive, and a small cluster of dead bees. I treated in August and now in November my bees are all dead. I looked for mites, but didn t see them so I didn t do any treatments. I am a natural beekeeper, so I did not treat them. Tracheal mites (Acarapis woodi) 1919 First found in Isle of Wight in English Channel US Honey Bee Act of 1922 prevented import 1984, tracheal mites found in USA. Extensive losses, breeding efforts, treatments, now hardly recognized Microscopic mite that lives in breathing tubes and is transmitted bee to bee. Control was chemicals (menthol) and disruption of the bee to bee transfer (grease) American foulbrood Paenibacillus larvae Easily spreads Contaminated comb, honey, clothing, tools & hands Kills pupa Ropes Clears milk Smells AFB How to diagnose Piece of comb 2 inches square Wrap in newspaper NOT in ziplock or plastic wrap Not molded or rotten Clinical signs ELISA tests Milk Test Laboratory o Aerobic culture on blood sagar o Also do sensitivity Kirby Bauer OTC and TYL o USDA or veterinary labs? Antibiotics to treat? Vegetative Stage - clinical disease Spore Stage able to spread disease, but not visible o Able to be inactive for several decades Prevention of AFB vs Cover Up? Radiation? Once a beekeeper gets on antibiotics, they are loathe to stop. Many books advocate antibiotics once or twice a year. 452

Shook swarm method AFB not extensive Shake all bees off frames into a box Burn Frames and scorch wood of hive bodies Place bees on new frames with bare foundation Bees use honey to power wax glands, excrete any spores outside instead of storing contaminated honey Feed antibiotic while building comb Variable success depending on severity. Most states do not allow this for AFB treatment. American foulbrood Is it best to suppress clinical signs by feeding antibiotics? Is it best to allow it to show up and then burn it to rid the apiary of spores? Overall, maybe 1% incidence Historically a lot higher 40% in New York State mid 1900s Reason most states have bee inspectors Many beekeepers and inspectors fear a massive outbreak once antibiotics are not available OTC. Others think this will help weed out bad beekeepers. European foulbrood Melissococcus plutonius Kills larvae Bacteria outcompetes larva for food and larva starves. Much more common than AFB Stress association o Forage availability o Humidity o Population of adults EFB diagnosis and control Clinical signs ELISA Culture o Frequent overgrowth with other bacteria New York State: o EFB is changing? Seeing it without characteristic upturning of larvae State inspector calling it AFB, lab confirms EFB Requeen Feed Increase population Feed Oxytetracycline o Resistance?? Shook swarm method is advocated in Europe Recurrence common, regulatory effort is minimal Chalk brood Ascosphaera apis Fungus Very common but not very severe problem Often spring time when low bee numbers per brood and have chilled brood Clears as colony strengthens and temperatures warm up Requeen if problem. 453

Nosema Caused by a single cell protozoan Two different types o Nosema apis Diarrhea disease of early spring May affect packages and weak colonies o Nosema ceranae o Newly recognized in US o Colony loss and dwindling of population o Bees tend not to demonstrate diarrhea o Significance not clear (2009) o Can be a severe problem (2010) Infects cells lining of midgut Less efficient use of protein and greater energy consumption due to poor absorption of nutrients A strong colony may not show symptoms Like other bee diseases, stressors make Nosema symptoms much worse. o Poor quality pollen o Cool damp weather o Small colonies o Failing queens o Long periods without cleansing flights Diagnosis Microscopy for spore counts Counts vary widely between bees and by season Healthy prolific colonies will experience decreasing Nosema levels Spore counts o Crush a number of bees with known water, Mix thoroughly o Count with hematocytometer o -ORo Smash single bee onto slide o Look for spores as high or low numbers o Problem is if 3 or more out of 10 have high numbers Diarrhea does not equal Nosema Normal feces High ash feed o Non-digestible feeds o Darker honey Lack of flying weather Amoeba? Treatment Nosema is a spore former Treatment may be necessary It is not appropriate to treat blindly though many do Little correlation between spore counts and illness Fumagilin???? o Nosema comes back worse after treatment? Reason to rotate frames out more often? Acetic acid or ozone fumigation of old combs? Fumagilin Antibiotic mixed in sugar syrup. Do not mix in hot syrup. 454

Apply to bees after or before honey flow, not during honey flow. No active NADA in the USA. Product imported from Canada and FDA CVM uses enforcement discretion to allow its import. Not on list of human medically important antibiotics, so still OTC Small hive beetles Aethina tumida Africa origination Egg, Larva, Pupa, Adult complete metamorphosis Pupa is outside hive in sandy soil Emerging as adult, can fly miles to find a bee hive. Fast movement Look for dark places to hide Can overwhelm weak hives Bees usually jail them by chasing to a hiding spot and not allowing them to leave Traps rely on this bees chase beetles into trap, oil in bottom, so beetles fall in and can t get out. Beetle larvae destroy honey Extract honey as soon as removed from hive Treatments and control Oil Traps Check Mite (coumaphos) hidden in piece of cardboard, mite accesses to hide, bees can t follow o Off the market currently Ground treatments GuardStar o Kill pupating mites o Beetles seek ground beyond the treatment Off label treatments as beekeepers experiment? Wax moths Strong populations of bees keep control. Dead or weak hive problem Brood combs destroyed Confused with Small Hive Beetle larvae Paradichlorobenzene control in stored equipment. Colony collapse disorder Colony of bees decreases from populous to brood with few adults and a queen without explanation Pesticides? Environment? Pathogens? Management? Neonicotinoids Fungicides Drought, mono-culture ag, yards lack of forage Nosema Varroa and viruses Poor management Pesticides Pickup pesticide from crop or exposed by spraying or forage on broken bags of pesticides o Large pile of dead bees in the hive/at hive entrance o Collect sample of bees, freeze them. o Call your state pesticide contact. o USDA pesticide screen ~$300 per sample. Neonicotinoids systemic pesticide o Safe for mammals o In pollen or nectar? o Sub-lethal effects on bees are an added stressor 455

Foreign animal diseases Tropilaelaps mites o Similar life cycle to Varroa mites o Asian Honey Bee mite Apis dorsata Brachypeplus basalis o Beetle similar to Small Hive Beetle o Now found in USA in commercial colonies on west coast Emerging diseases? Sepsis and Hemocyte Loss in Honey Bees (Apis mellifera) Infected with Serratia marcescens Strain Sicaria Citation: Burritt NL, Foss NJ, Neeno-Eckwall EC, Church JO, Hilger AM, Hildebrand JA, et al. (2016) Sepsis and Hemocyte Loss in Honey Bees (Apis mellifera) Infected with Serratia marcescens Strain Sicaria. PLoS ONE 11(12): e0167752. doi:10.1371/journal.pone.0167752 456

Honey Bee Veterinary Services Christopher Cripps, DVM Betterbee Grennwich, NY Veterinarians and bees FDA now mandating veterinary involvement for antibiotic use Veterinarians need to jump up and seize the opportunity! USDA does not look to veterinary involvement Bees need o Preventive Care o Parasite Control o Disease Diagnosis o Disease Treatment o Beekeeper education Veterinarian in the apiary For a valid VCPR, the veterinarian needs to visit the apiary (bee yard) Moving bees to the veterinary office is difficult Dead hives may be brought to the office? Smartphones and pictures? o What can you learn on site? o How strong are the hives o Where are they kept o Water available? o How does the beekeeper keep the bees? o How does the beekeeper act around the bees? o Unrecognized conditions in other hives? o When visiting, you should have a veil that will protect your face and head You should have a clean hive tool and smoker It may be advisable to have the beekeeper provide these for you You should not wear leather gloves. We find Nitrile milker gloves work well Have sampling supplies available plastic bags, bee sampling bottles, newspaper Walk and conduct yourself in a slow deliberate manner to avoid upsetting bees. Establish a list to examine so you can do a thorough physical exam Clean up when you leave bucket of water, chlorinated cleaner like Comet, steel wool, torch to heat hive tool History why are we here? What have you done? External examination environment, hive appearance, behavior of bees at entrances, soil Internal examination behavior of bees, size of hive, spacing of frames, smells, presence of other insects, abnormalities of the bees Internal examination brood frames, color, type of brood, pollen, honey stores, wax moths, beetles, larval diseases, brood pattern Internal examination what is on the bottom board or sticky board Commercial beekeepers Sign a prescription or VFD so I can do what I always do. Disease sampling Disease recognition Disease management Protocols for when to use treatments All the stuff listed for Hobbyists Hobbyist beekeepers Education How to work bees 457

What is normal? What caused this problem? Do I have a problem? What do I do about it? Veterinarian in the apiary Veterinarians are highly trained in physiology, anatomy, disease, diagnosing, treatment and preventing the FDA recognizes this and is why they require a veterinarian before antibiotics are used. Many beekeepers are older and have done bees for a lot of years. They can still learn about bees! People earn PhDs in beekeeping There are a lot of stories in beekeeping that may or may not be true. They also may or may not apply to the situation at hand. More is missed for not looking than not knowing Thomas McCrae Veterinarian in the apiary Subjective Objective Assessment Plan Works as well in the apiary as it would in the hospital Medical records prove VCPR if any issues with VFD or prescriptions Antibiotics in honey bees Commercial beekeepers o Move bees into crops for pollination o Poor biosecurity o High disease exposure o Feed antibiotics to kill whatever they can to prevent bringing home disease Commercial beekeepers o Weak hive treatments and prevention o Queen mating hives Any beekeeper o Prevention of American Foulbrood o Should we? Are we just covering up previous bad management? Veterinary Client (Beekeeper) Patient (beehive) Relationship o Physical presence at apiary o Yearly? Every 6 months? o Personally acquainted with husbandry o Willing to accept responsibility for medical management and follow up o Beekeeper willing to follow directions There is a lot of non-labeled product going into beehives. Can you control this behavior? 3 antibiotics with Honey Bee labels o Oxytetracylcine, Tylosin, Lincomycin All are given to bees by mixing powdered antibiotic with powdered sugar or mixing in a grease patty Apply 2 Tablespoons to hive by spreading around outside of hive. Do not apply to brood Oxytet has VFD and RX forms Others have only RX forms Residue avoidance Antibiotics will appear in honey Honey is tested for antibiotic by some packers No reporting requirement to regulatory bodies o Milk is different from honey 458

Beekeepers feed antibiotics before any honey being collected or after removed honey for human consumption o Honey for humans usually collected late spring though early fall. ELISA tests available for on farm or in plant testing Antibiotics in honey bees Prior to writing VFD or RX, do I need a diagnosis? Prevention, Control, Treatment o FDA terms without definitive definitions o Control means there is disease and you are keeping it from spreading in others that you treat What is the quality of a diagnosis? o Physical signs o In house/hive side testing o 3rd party laboratory testing Hive side or in clinic ELISA tests About $13 each AFB or EFB Highly purified monoclonal antibodies used to detect antigen of bacteria Mix samples in bottle of fluid, mix well, place sample on well, wait 5 minutes Line on C=control T=test Culturing honey bee bacteria Paenibacillus larvae o Grey white colonies on agar. Orange possible also o Spore forming and motile rods o G+, but difficult to stain o Catalase -, esculin + o Also consider wet mount. Only bee pathogen that shows Brownian motion. Melissococcus plutonius o Small white, shiny, well defined colonies o Non motile cocci in pairs or chains o Difficult to grow, other associated bacteria also grow USDA lab Beltsville MD Brownian motion on wet mount Culture on blood agar o Maybe heat shock AFB makes a spore AFB fluorescent under UV light Smell Kirby Bauer Disk for resistance 5microgram Oxytetracycline disk Most isolates clear at least 50mm zone Less than 50mm zone is resistant Dead hive necropsy Lots of causes of dead hives. Diagnosis of cause of death or ruling out a cause may help beekeeper apply limited resources to prevention Starvation Varroa Mites American Foulbrood Freezing The Necropsy Are any bees present 459

Honey present? Bees present? Organized cluster? Brood present? Fecal staining? Testing Using deadout resources If AFB and Nosema can be ruled out combs can be reused Honey can be fed Combs will greatly aid packages or nucs Moldy combs Secure resources before robbing/wax moths Many beekeepers will destroy these valuable resources in case a low incidence disease caused the deadout. Parasitic mite syndrome Guanine deposits = Mite poop European foulbrood Curled larvae Out competed for food Spotty brood patterns Culture ELISA Confused with AFB It is a great service to diagnose correctly as this does not need to be burned! Diagnosing conditions Laying worker All drone brood Worker did not get inseminated so only lays haploid eggs and cannot reach bottom of cells Treatment o Add a queen via a nuc o Combine with other queenright hive My hive swarmed two weeks ago, now there are no eggs or queen, how do I introduce a new queen? Maybe a queen is not needed. By knowing normal bee biology, we know swarm leaves between a week prior to new queen emerging and the day of emergence, it takes her a week to mature, then she goes on another week of mating flights. At 2 weeks post swarm, no eggs or queen normally. New queen may start laying soon. Chilled brood Expand brood nest Cold weather, so bees cluster and cannot keep brood warm Beekeeper reverses hives and splits brood A lot of resources for bees to clean this up Overheated bees Bees are a heat producer Need ventilation Nucs or packages or hives In transport, usually netted, not locked in Queen s sperm storage may kill future fertility. Euthanasia of bees Sprays require contact and are effective 460

Sprays are short lived Sprays tend not to get carried into a hive If bees are inside a cavity, sprays won t work. You are better to open the cavity and remove the combs and bees. Overturned trucks, ground bees (yellow jackets) Soapy hot water Firefighting foam Surfactants that break the surface tension and allow liquid to enter trachea of bees. Microscopy Tracheal Mites Nosema Veterinarians and beekeepers Veterinarians are highly trained biologists that can systematically examine a system for abnormalities. Veterinarians are highly trained in disease diagnosis, treatment and prevention More is missed for not looking than not knowing People earn PhDs in beekeeping There is a lot of superstition out there. The FDA is looking for veterinarians to bring the science. 461

Honey Bees and Veterinarians Christopher Cripps, DVM Betterbee Grennwich, NY Chris Cripps Started beekeeping with the Boy Scout Beekeeping Merit Badge in mid 80 s in New Hampshire Cornell classes in Bee Biology with labs Ohio DVM OSU o Inspector for 2 counties (Columbus area) o Make appointment with owners, look over hives for diseases, take samples for lab as needed, report health status to owner and recommend any treatment needed. o Moved 6 hives to a horse farm in Ohio from NH New York Greenwich o Arrived in 1995 with 6 hives, all my stuff, and $10 in the back of a U-haul o Kept up to 12 hives, sold honey, moved bees for pollination o Worked as dairy veterinarian at Battenkill Veterinary Bovine September 2012 bought Betterbee (bee supply business) and left veterinary practice??? Now partners with veterinarians Joe Cali and Jack Rath and 2 others. Betterbee Beekeeping supply business that sells bee hives, frames, foundation, extractors, queen rearing supplies, fencing, honey bottles, bears, caps, books, classes Beekeepers Serving Beekeepers Teach beginner & intermediate beekeeper classes Answer questions from customers Supply scientific information about beekeeping Honey bee veterinary practices With 3 veterinarians, I thought we had the largest group of veterinarians in honey bees Visited Wilbanks Apiary in Claxton, Georgia 7 veterinarians worked there one year- Argentinians Honey bees Veterinary curricula in many countries include honey bees USA and Canadian schools do not include bees Don t worry, you did not miss it, it is not offered New antibiotic labeling and registration requires a veterinarian to write an order for any antibiotic to be fed to animals Honey Bees are Food Producing Animals that have been fed antibiotics Only insect listed as food producing animal How did it work previously? Antibiotics with labels for Honey Bees: Oxytetracycline Tylosin Lincomycin Indications: for the control of American or European Foulbrood Self-reporting survey 2015 of ~5000 beekeepers about 1/14 reported antibiotic use They averaged ~900 hives Honey bees and antibiotics: How did it work before? Commercial beekeepers feed antibiotics to prevent disease o Mostly oxytetracycline mixed in sugar that has been over the counter Little understanding of bacteriology o Oxytet to start o Tylosin used if oxytet doesn t work o Lincomycin seldom used Gather hives in highly populated areas for pollination California almonds, Maine blueberries 462

o Cesspools of disease that cannot be avoided because the pay is so good and may mean the difference between having a profit or loss for the year Honey bees and antibiotics: How does it work now? Prior to transportation, must obtain a certificate of inspection Issued by state apiarist Bee inspectors visit apiaries, remove brood from a percent of hives (~10%). Submit samples to USDA if any concerns or issue health certificate if no concerns Veterinary epidemiologist might help what percentage of frames should be inspected to find disease assuming 1% prevalence? Positive American Foulbrood (AFB) may invoke state-mandated burning of affected hives Long lasting spores are contagious to other hives within 3 mile radius Burning beehives worth $500 each makes beekeepers wary of outside inspection Beekeepers like other food producers Frugal Expect value for money spent Veterinarians sell what? o Knowledge and disease control techniques o Signature on antibiotic orders that they did not previously need to have If you come to the table to offer services, make sure you offer value o Get Educated! Important points in bee biology Complete Metamorphosis o Eggs, Larvae, Pupae, Adult o 1 to 2,000 eggs per day Worker egg to adult o 21 days Queen egg to adult o 16.5 days Drone egg to adult o 24 days o Important for Varroa Mites! 3 distinct castes of bees: o Queen (diploid) fertile female that lays eggs o Workers (diploid) infertile females that care for young (brood), gather food, clean and defend hive. o Drones (haploid) males that breed queens Bee sex and genetics Queen emerges from cell After a week of maturing in the hive, starts mating flights Mates with 8-20+ drones over next week Never mates again Does not mate with drones in hive Sperm must live many years in queen s spermathecal Langstroth 1852 Bee Space = 3/8 inch Movable Frame Hives Wax production Carbohydrate drives glands on abdomen in young workers Feeding sugar can push more wax production Wax used to make all comb Paper used by wasps and hornet, not bees! 463

Lipophilic chemicals persist in wax o Chronic pesticide exposure Lots of brood leads to a lot of bees which lead to a lot of honey Package of bees is 3 lbs o Common way to start Contains ~10,000 bees Colony that survived winter will be small and grow through the year Starter bee colonies Swarm Package Nucleus Hive Honey bee food storage Protein = pollen Need variety of pollens for a balanced Amino Acid profile Carbohydrates = nectar Perhaps 20% solids converted to 17% water Invertase, dehydration Mixture, ferment = bee bread Do fungicides hurt bees? Honey bees, directions and food Drifting Bees make orientation flights to determine where their hive is before foraging. Bees may return to wrong hive if many in same area Welcomed if they have food Robbing Strong hives defend themselves from robbing Weak hives robbed by strong hives Drifting and Robbing can lead to disease spread Winter time Honey bees keep cluster heated throughout the winter ~95deg F when raising brood in January Hornets and wasps overwinter on or underground as individuals Bees don t defecate in hive Process honey to keep hive warm Need to fly to defecate Cleansing Flights Once below 50⁰F, cannot operate muscles, fall out of air Protective clothing Veils Blinking attracts bees Gloves o Leather cannot be cleaned o Nitrile gloves Coveralls and jackets with incorporated veils Smokers Calms bees Burn many different Materials Light with a propane torch 464

Store safely if driving between apiaries Clean bellows well to avoid disease spread Bee stings Stinger is modified ovipositor o Drones do not sting Worker sting barbed o Rip insides of bee out o Only one sting then dead Swelling normal reaction More times stung, less swelling Anaphylaxis possible o Epinephrine o Diphenhydramine Beekeeper behaviors Work bees in a calm manner Sudden jarring motions elicit defensive behavior Falling barometric pressure, rain increase bee defensiveness Black colored clothing not good Wool or fleece not good Bees get legs caught Suit of armor and go hard at bees Wear nothing but veil and be gentle Crushing bees can lead to more disease issues Veterinarians and beekeepers Diseases for which antibiotics may be used American Foulbrood European Foulbrood Other disease treatments are Over the Counter EPA regulated mite treatments Non-medically important antibiotic To order antibiotics: 2 choices Prescription Requires VCPR in most states Typical form used in clinics Oxytetracycline, tylosin or lincomycin as water soluble forms Send to licensed pharmacist or give to client ELDU is allowed Keep records per state law Expiration max??? Veterinary feed directive Requires VCPR New to most veterinarians Oxytetracycline only Tylosin, Lincomycin have no VFD approval for bees Send to licensed medicated feed mill or distributor ELDU prohibited, but not enforced. Keep original records for 2 plus years Expiration max 6 months Extra label drug use Labeled antibiotics 465

For the CONTROL of o Indicates there is a diagnosis of AFB or EFB o Treating other hives to prevent spread Most antibiotic use will be for PREVENTION or TREATMENT o No antibiotics labeled in honey bees for these indications VFD cannot be written for ELDU but RX can o New FDA announcement allows ELDU in minor species with some restrictions. Check these out first! o Still too limiting to help beekeepers??? Honey bee prescription issues Identification Beehives tend not to be individually identified o No state tags Apiaries identified by address Most states say you must personally visit the apiary for a valid VCPR Follow up may be by other means once valid VCPR is established o Telephone o Pictures/Email AVMA PLIT will acknowledge your work with bees Products available Feed mills may make product for VFD fulfillment Need not be antibiotic licensed feed mill Type A medicated article (Terramycin) comes in bag for 8000 treatments Prescription product of oxytetracycline is in much smaller packets Should we be using antibiotics? European foulbrood o Can be a problem and antibiotics can be convenient treatment o Small queen rearing hives American Foulbrood o May cover up the clinical signs and hide disease o Still contagious o Transfer disease to people buying used equipment o Many people think it should not be used o Europe prohibits Veterinarians and bees Who is in a better place to provide this than veterinarians? To provide those service, gain knowledge If you understand the disease and its nuances, the beekeeper may view you as a source of knowledge rather than another bill Join the Honey Bee Veterinary o New website to help beekeepers find veterinarians 466