Naslov
UTJECAJ DODATAKA HRANI NA STUPANJ INVAZIJE MIKROSPORIDIJOM Nosema ceranae, BIOKEMIJSKE I HISTOKEMIJSKE POKAZATELJE U MEDONOSNE PČELE (Apis mellifera)
(INFLUENCE OF FOOD SUPPLEMENTS ON DEGREE OF INFECTION BY MICROSPORIDIUM Nosema ceranae, BIOCHEMICAL AND HISTOCHEMICAL INDICATORS IN THE HONEYBEE (Apis mellifera))

Autori
Ribarić, Jasna

Vrsta, podvrsta i kategorija rada
Ocjenski radovi, doktorska disertacija

Fakultet
Veterinarski fakultet

Mjesto
Zagreb

Datum
31.03

Godina
2016

Stranica
100

Mentor
Tlak Gajger, Ivana ; Nejedli, Srebrenka

Ključne riječi
Apis mellifera; nozemoza tipa C; dodatci hrani za pčele; NozevitPlus; EkoZeoPet; BEEWELL AminoPlus
(Apis mellifera; nosemosis type C; feed additives; NozevitPlus; EkoZeoPet; BEEWELL AminoPlus)

Sažetak
Nametnička bolest nozemoza tipa C uzrokovana mikrosporidijom Nosema ceranae jedna je od najraširenijih bolesti medonosnih pčela u svijetu. Bolest uzrokuje naglu depopulaciju odraslih pčela i moguće propadanje invadiranih pčelinjih zajednica čime značajno narušava održivost i biološku ravnotežu prirodnih ekosustava, i profitabilnost pčelarenja kao poljoprivredne grane. Obzirom na moguće gospodarske štete, brzo umnažanje spora uzročnika N. ceranae te ne dozvoljenu uporabu antibiotika u pčelarstvu Europske unije, cilj ovog istraživanja bio je utvrditi učinkovitost višekratne primjene dodataka hrani za pčele biljnog sastava NozevitPlus, mineralnog sastava EkoZeoPet i proteinsko-vitaminskog sastava BEEWELL AminoPlus na stupanj invadiranosti pčelinjih zajednica mikrosporidijom N. ceranae, kao i biokemijske (glukozu, ukupne bjelančevine i masti u ličinkama) i histokemijske (prisutnost i distribuciju više vrsta mukopolisaharida i aktivnost proteolitičkih enzima u srednjem crijevu odraslih pčela) pokazatelje te jačinu pčelinjih zajednica. Rezultati ovog istraživanja pokazuju da primjena sva tri dodatka hrani za pčele pozitivno utječe na smanjenje broja spora N. ceranae i jačinu pčelinjih zajednica, te se stoga mogu preporučiti kao alternativni način kontroliranja i liječenja ove bolesti.

Izvorni jezik
Hrvatski

Znanstvena područja
Veterinarska medicina

Napomena
Extended abstract Honeybees (Apis mellifera) are the predominant and most economically important group of pollinators in most geographical regions. Honey bee colony losses and declines, which are the result of multiple stressors, working independently or synergistically to impact honey bee health, have caused much concern worldwide. These losses threaten the economic stability of commercial beekeeping and have serious implications to pollination services for both cultivated and wild plants. Nosemosis type C, caused by Nosema ceranae infection is now considered a major health problem affecting beekeeping worldwide. The endoparasitic fungal infections of N. ceranae adversely affect honey bee colony health and can result in complete colony collapse. The only widely used treatment for nosemosis type C, fumagillin, is banned in the EU. Moreover, prolonged treatment with fumagillin may contribute to drug resistance and may exacerbate N. ceranae infection rather than supress it. As treatments for N. ceranae are currently unavailable in many countries, one of the biggest challenges being faced by the scientists are further studies of potential treatments or beekeeping techniques which are urgently required to combat the rapid spread of this dangerous emerging disease. In the present study the effect of repeated applications of three food supplements NozevitPlus, EkoZeoPet and BEEWELL AminoPlus (including herbs, minerals and protein ingredients) have been tested to control N. ceranae infection in honey bee colonies. The experiment was conducted during July and August of 2014. During the experiment 30 honeybee colonies of the same apiary, naturally infected with Nosema spp., were randomly divided in three experimental and control group of honeybee colonies. The experimental groups were treated with sugar syrup supplemented with food supplements NozevitPlus, EkoZeoPet and BEEWELL AminoPlus. None of the colonies showed symptoms of other disease and none received any other treatment. The adult honeybee and larval samples were collected on 1st, 10th, 20th and 30th day of feeding. To evaluate Nosema spp. infection in the colonies, samples of forager bees and house bees were collected at noon. The spore concentration was determined by counting with a Bürker-Türk hematocytometer chamber and the Nosema species was determined by multiplex PCR. The adult bee population and brood production were evaluated throughout the assay by analyzing the frames covered by bees and quantifying the number of brood cells. Total protein concentrations were determined using a modified Lowry Protein Assay in suspension of larvae tissue. Glucose in larvae samples was measured colorimetrically using this commercially available Glucose Assay Kit and the total fat content was done by gas chromatography. Extraction of the fat from previously homogenised larvae was performed by the method of Bligh and Dyer. Degreased cuts of the midgut were stained according to the HE method in order to determine the general morphological characteristics of the tissue, and stained with special stains for determination of enzymes activity. In addition to examining the histopathological structure as well as content and distribution of mucosubstances and histochemical activity of aminopeptidase a variety of stains were used to describe neutral, acid and sulphate mucopolysaccharides (periodic acid-Schiff [PAS], alcian blue pH 1.0 [AB1], alcian blue pH 2.5 [AB2.5], toluidine blue [TB]). All composite forager bee and house bee samples analysed tested positive for N. ceranae infection. Infection by N. apis was not detected. The study demonstrated that the disease was not cured, but all three food supplements administrated via sugar syrup reduced development of microsporidium N. ceranae in honeybee midgut (NozevitPlus: 11, 35% on 10. day, 23, 56% on 20. day and 50, 54% on 30. day ; EkoZeoPet: 15, 62% on 10. day, 7, 68% on 20. day and 35, 51% on 30. day ; BEEWELL Aminoplus: 21, 33% on 10. day, 26, 89% on 20. day and 57, 31% on 30. day). Mean of total protein concentration observed in this study ranged for NozevitPlus between 0, 094 and 0, 137, for EkoZeoPet between 0, 088 and 0, 120 and for BEEWELL AminoPlus between 0, 087 and 0, 114 (mg/mg larvae tissue). The glucose mean values concentrations ranges were as follows: NozevitPlus 9, 25-10, 58, EkoZeoPet 9, 24-10, 57, BEEWELL AminoPlus 9, 12-10, 13 (mg/mg larvae tissue) and the total lipids mean values concentrations ranges were as follows: NozevitPlus 0, 031-0, 034, EkoZeoPet 0, 033-0, 036, BEEWELL AminoPlus 0, 023-0, 028 (mg/mg larvae tissue). Among these results isn’t found statistically significant difference, as well as between experimental and control group of honeybee colonies. These results demonstrate the efficacy of these food supplements in reducing spores found in the honeybee mid-gut over a short period of time and suggest that they may limit mortality in bees and be beneficial in nosemosis type C treatment. Further field tests are needed to provide stronger evidence on the potential of these three food supplements.

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