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The Investigation of Honey Bee Diseases After Colony Losses in Hatay and Adana Provinces of Turkey

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The Investigation of Honey Bee Diseases After Colony Losses in Hatay and Adana Provinces of Turkey MELLIFERA 10-20:24-31 (2010) HARUM 24 RESEARCH ARTICLE THE INVESTIGATION OF HONEY BEE DISEASES AFTER COLONY LOSSES IN HATAY AND ADANA PROVINCES OF TURKEY TÜRKİYE, ADANA-HATAY İLLERİNDEKİ KOLONİ KAYIPLARININ ARDINDAN BAL ARISI HASTALIKLARININ İNCELENMESİ Aygün Yalçınkaya* • Nevin Keskin* Summary: Sudden colony losses occurred in Turkey, mostly in Hatay and Adana region in the year of 2007. After this event, all of our laboratory experiments were focused on this region. The aim of the study was to investigate the situation of honey bee diseases in Hatay- Adana region and to determine probable causes of colony losses. In this research, 97 and 88 honey bee brood combs were collected from Adana and Hatay during the spring and autumn field works. The total numbers of investigated adult honey bee were 3880 from Adana and 3520 from Hatay. All debris and adult honey bee samples were investigated under dissection microscope for diagnosis of Varroasis. Spore counting method was used for Nosemo- sis. “Guanine visualization” and “dissection” methods were used for diagnosis of Acarapiasis. Larval samples were inoculated to selective media for determination of American and European Foulbrood. For bacterial identification, biochemical tests used and Gram- stained slides were investigated under light microscope. As a result of laboratory analyses, Varroasis was determined in the ratio of 98% and high level Nosemosis was determined in the ratio of 12.97%. As a result of diagnostic tests that were applied to all honey bee samples, American Foulbrood and European Foulbrood were detected at the rate of 29% an 19% respectively. The ratio of the healthy bees was 52%. Consequently, we saw that parasitic and microbial diseases exist and they are prevalent in this region but the level of diseases showed that not only a single disease but also the combination of diseases and other factors could cause severe colony losses. Keywords: American Foulbrood, European Foulbrood, Varroasis, Nosemosis, Acarapiasis, Colony Losses Özet: Türkiye’de 2007 yılında, çoğunlukla Hatay ve Adana bölgesinde ani koloni kayıpları meydana gelmiştir. Bu kayıplardan sonra laboratuar çalışmalarımız bu bölge üzerinde yoğunlaşmıştır. Çalışmanın amacı, Adana-Hatay bölgesindeki arı hastalıklarını araştırmak ve koloni kayıplarının olası sebeplerini belirlemektir. Bu çalışmada, ilkbahar ve sonbahar arazi çalışmalarında Adana ve Hatay’dan 97 ve 88 yavrulu petek toplanmıştır. Adana’dan 3880, Hatay’dan 3520 adet ergin bal arısı incelenmiştir. Tüm deb- ris ve ergin bal arısı örnekleri Varroasis tanısı için diseksiyon mikroskobu altında incelenmiş; Nosemosis için spor sayma yöntemi kullanılmıştır Acarapiasis tanısı için Guanin görüntüleme ve diseksiyon yöntem- leri bir arada uygulanmıştır. Amerikan ve Avrupa yavru çürüklüğü hastalıklarının belirlenmesi amacıy- la larva örneklerinden, seçici-ayırdedici besiyerlerine inokulasyonu yapılmıştır. Bakteriyel tanımlama yapılması amacıyla biyokimyasal testler kullanılmış; Gram boyama yapılan preparatlar ışık mikroskobu altında incelenmiştir. Laboratuar analizleri sonucunda Varroasis % 98, yüksek seviyedeki Nosema % 12,97 oranında tespit edilmiştir. Tüm bal arısı örneklerine uygulanan tanısal testler sonucunda Amerikan Yavru Çürüklüğü % 29 ve Avrupa Yavru Çürüklüğü % 19 olarak saptanmıştır. Sağlıklı arıların oranı ise % 52 olarak tespit edilmiştir. Sonuç olarak, bölgede paraziter ve mikrobiyal hastalıkların yaygın olarak bulunduğu görülmüş ancak hiç bir hastalığın tek başına büyük çaplı kayba neden olmayacağı, koloni koyıplarında arı hastalıkları ve diğer faktörlerin bir arada etkili olduğu kanaatine varılmıştır. Anahtar kelimeler: Amerikan Yavru Çürüklüğü, Avrupa Yavru Çürüklüğü, Varroasis, Nosemosis, Aca- rapiasis, Koloni Kayıpları *Hacettepe University, Department of Biology, Bee Health Lab., Beytepe Campus, 06800, Ankara/Turkey e-mail: [email protected] 25 Introduction The situation of Nosemosis still is not clear worldwide Honey bees have important roles not only in the pro- and further researches are necessary. duction of honey and bee products, but also pollina- ting of agricultural crops. Colony depletion and dise- American foulbrood (AFB) is caused by the bacterium ase outbreak in managed honey bees are not unusual. Paenibacillus larvae and European foulbrood (EFB) However, serious colony losses and decline in bee po- is caused by the bacterium Melissococcus pluton pulations have been reported in recent years and the (Bailey,1983). Both species are Gram-positive bacte- term CCD (Colony Collapse Disorder) was first used ria. Melissococcus pluton is a chain, single or double all over the world in the year of 2006 (vanEngelsdorp formed coccus. Several other bacteria like Acromo- et al., 2007). This decline in pollinators is a global bacter euridae, Enterococcus faecalis, Paenibacillus threat to biodiversity. Therefore honey bee health has alvei, Brevibacillus laterosporus maybe associated a great impact on economy and biodiversity worldwi- with EFB (Forsgren, 2010). Paenibacillus larvae is a de (Jaffé et al., 2009). spore forming, Gram-positive rod-shaped bacterium. AFB is common bacterial disease affecting apiculture There are a lot of parasites and pathogen microor- worldwide. It is included in the Notifiable Disease list ganisms associated with honey bees affecting adult by the World Organization for Animal Health (OIE). bees and brood. Parasitic mite Varroa destructor is AFB is difficult to be managed by apiculturists be- the most damaging honey bee parasite in the world cause the pathogen produces environmentally stable (Rosenkronz et al, 2010). Mite infection which causes spores which are very virulent and infectious after dramatic colony losses have repeatedly occurred in decades, as well as resistant to heat, to desiccation affected countries. Adult Varroa mites feed on adult and chemical disinfectants and it spreads easily and bees, but reproduction depends on bee brood. Both the rapidly within a colony, among colonies in an api- adult female and her offspring feed on pupae, where ary and between apiaries (Ashiralieva and Genersch, they can cause damage by infestation of hemolymph, 2006; Fries et al. 2006) resulting in severe nutritional deficits for the develo- ping bee (Bowen-Walker and Gunn, 2001, Duay et al., It is aimed to search all kinds of honey bee diseases 2003, Garedew 2004). Varroa mites can transmit se- in Hatay and Adana provinces in this study. Because, veral viruses which cause severe damages on bees and Hatay and Adana provinces serve as a resource for cit- colony losses (Chen et al., 2006). Acarapis woodi is rus and cotton honey, and have important role in Tur- another mite related to honey bees. Tracheal mite Aca- kish Beekeeping as wintering location for migratory rapis woodi damages tracheal walls mechanically and beekeepers. Colony losses in these provinces caused causes blockage on tracheas by its metabolic products great impact on Turkish Beekeeping industry. (Erickson et al.,1999) Nosema apis and Nosema ceranae are intracellular Material and Methods microsporidian parasites infecting the midgut epit- Collection of Samples helial cells of adult honeybees (Forsgren and Fries, In this study adult and brood honey bee samples were 2010). Nosema ceranae is more virulent than N. apis collected from Adana and Hatay provinces at early and it is replacing with Nosema apis in most count- spring and late fall of 2006-2007 season, in coopera- ries and Nosemosis is worldwide (Higes et al., 2007, tion with Adana and Hatay Beekeeping Associations Klee et al., 2007, Paxton et al., 2007). It is known to (Table 1). All samples were examined for parasitic and be correlated with reduced lifespan of individual bees, microbial diseases. Field studies were carried out in reduced performance of colonies and increased winter Aladağ, Ceyhan, İmamoğlu, Karataş, Karaisalı, Ko- mortality (Fries, 1993). On the other hand, some pub- zan, Pozantı, Saimbeyli, Seyhan, Tufanbeyli, Yumur- lications emphasized that Nosema ceranae is not rep- talık districts of Adana province and Antakya, Belen, lacing with Nosema apis and their virulence is not so Dörtyol, Erzin, Hassa, İskenderun, Kırıkhan, Rey- different in some countries (Forsgren and Fries, 2010). hanlı, Samandağ, Yayladağ districts of Hatay provin- MELLIFERA 26 ce. Live adult bee samples and brood were collected 0.2x0.2x0.1=0.004 mm cube 9x0.004= 0.036mm cube directly from colonies and brought to the laboratories A/0.036= mm cube x 1000= number of spore/ml in cold conditions (4 0C). Examination of Brood Honey Bee Samples Examination of Adult Honey Bee Samples Brood foundation was examined for clinical signs. All adult bees were firstly investigated under dissec- Diseased larva appears moist and darkened, cell cap- tion method for Varroasis and other symptoms like ping is concave and possibly punctured in foulbrood swollen abdomen and hair losses. Varroa mites have diseases. For isolation of bacteria, larval samples were been detected on adult bees and also in debris. For inoculated firstly to Nutrient Broth, Brain Heart In- the detection of the tracheal mite Acarapis woodi, two fusion Broth and then from broth cultures to Brain different methods were used: Dissection method and Heart Infusion Agar with Nalidixic acid and MYPG Guanin visualization method. Dissection method was Agar (Mueller-Hinton broth, yeast extract, potassi- applied due to OIE manual. Adult bees were secured um phosphate, glucose and pyruvate). Cultures were on their backs. Their heads and forelegs were remo- incubated
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