Bihejvioralna odbrana sjeničko-pešterskog ekotipa medonosne pčele od krpelja Varroa destructor

Two behaviours of honey bees, hygienic and grooming, are mechanisms of defense against brood diseases and parasitic mites, including Varroa destructor. Apis mellifera colonies remove the worker brood infested with Varroa destructor mites from the nest (hygienic behaviour), and groom the mites off other adult bees (grooming behaviour). In this study hygienic and grooming behaviours of Sjeničko-Pešterski honey bee ecotype were analysed in 440 honey bee colonies from 11 localities in the region of Sjeničko-Pešterski plateau, Podpešterje, Golija Mt. and Rogozna ML At each locality 40 honey bee colonies were investigated: 10 potent colonies with one-year old queen, 10 potent colonies with two-year old queen, 10 medium potent and 10 weak honey bee colonies. Hygienic behaviour was expressed in a range from 95.12% to 99.50% in potent honey bee colonies with one-year old and two-year old queens. Statistically highly significant (p lt 0.01) differences were registered among the analysed honey bee colonies at the investigated region, in favour of the potent honey bee colonies, compared to the medium potent and weak colonies. Also, statistically highly significant (p lt 0.01) differences were recorded between potent colonies with one-year old queens and colonies with two-year old queens, in favour of the colonies with one-year old queens. In general, investigated colonies belong to a category of the so called "hygienic colonies", as the efficiency of elimination of damaged pupae amounted to 91.50%. Grooming behaviour of Sjeničko-Pešterski honey bee ecotype potentially exists, but its significance cannot be discussed as, on the whole, investigated colonies showed potential of 34,04%. Our results point to an indisputable relationship between analysed behaviours and the strength of honey bee colonies: hygienic behaviour is more expressed in potent colonies (from 95.12% to 99.50%) regardless of queen age; grooming behaviour was expressed only in potent honey bee colonies with one-year old queen at all 11 localities, where the number of damaged mites ranged from 36,05% to 39,61%. The damaged mites were separated into six categories. The most frequent category of damage was damaged legs (53.38% in potent colonies with one-year old queens and 52.02% in potent colonies with two-year old queens). The potent honey bee colonies from the investigated region especially with one-year old queen, could be used for highly selected breeds improving and queens rearing.Higijensko i negovateljsko ponašanje predstavljaju dva mehanizma odbrane od bolesti legla i parazitskih krpelja, uključujući Varroa destructor. Društva Apis mellifera uklanjaju iz satnih ćelija radiličko leglo infestirano krpeljima Varroa destructor (higijensko ponašanje) i skidaju krpelje sa adultnih pčela (negovateljsko ponašanje). U ovom radu, obavljena je analiza higijenskog i negovateljskog ponašanja pčela sjeničko-pešterskog ekotipa kod ukupno 440 pčelinjih društava sa 11 lokaliteta Sjeničko-Pešterske visoravni Podpešterja, Golije i Rogozne. Na svakom lokalitetu ispitivano je 40 pčelinjih društava: 10 jakih društava sa jednogodišnjom maticom, 10 jakih društava sa dvogodišnjom maticom, 10 srednje jakih i 10 slabih pčelinjih zajednica. Higijensko ponašanje bilo je ispoljeno u opsegu od 95,12% do 99,50% kod jakih pčelinjih zajednica sa jednogodišnjim i dvogodišnjim maticama. Između ispitivanih pčelinjih društava pomenutog područja registrovane su statistički visoko značajne razlike (p lt 0.01) u ispoljenosti higijenskog ponašanja u korist jakih pčelinjih zajednica u odnosu na srednje jake i slabe. Takođe, visoko značajne razlike (p lt 0.01) u ispoljenosti ove osobine zabeležene su i između pčelinjih zajednica sa jednogodišnjim i dvogodišnjim maticama, u korist pčelinjih zajednica sa jednogodišnjim maticama. Generalno, pčelinja društva sa analiziranog područja Sjeničko-Pešterske visoravni, Podpešterja, Golije i Rogozne pripadaju kategoriji higijenskih društava, obzirom da je prosečna efikasnost eliminacije oštećenih lutki svih zajednica iznosila 91,50%. Negovateljsko ponašanje kod pčela sjeničko-pešterskog ekotipa potencijalno postoji, ali se ne može govoriti o izraženosti ove osobine, obzirom da je globalna sposobnost eliminacije krpelja sa oštećenjima u ukupnom broju odbačenih krpelja svih ispitivanih pčelinjih zajednica bila 34,04%. Ovi rezultati ukazuju na postojanje neosporne povezanosti analiziranih oblika ponašanja i jačine pčelinje zajednice: higijensko ponašanje je vise ispoljeno kod jakih društava (od 95.12% do 99,50%) nezavisno od starosti matice, dok je negovateljsko ponašanje bilo izraženo samo kod pčelinjih zajednica iz kategorije jakih društava sa jednogodišnjom maticom na svim lokalitetima (od 36,05% do 39,61 % oštećenih krpelja u odnosu na celokupan uzorak). Oštećeni krpelji su podeljeni u 6 kategorija. Najčešći tip oštećenja predstavljaju oštećene noge (53.38% u jakim društvima sa jednogodišnjom maticom i 52.02% kod jakih društava sa dvogodišnjom maticom). Jaka pčelinja društva sa ispitivanog područja, i to naročito ona sa jednogodišnjom maticom, mogu koristiti za poboljšanje visoko selekcionisanih sojeva, kao i u proizvodnji selekcionisanih matica sa izraženim higijenskim i negovateljskim ponašanjem

Heritabilnost higijenskog ponašanja sive medonosne pčele (Apis mellifera carnica)

Breeding grey A. m. carnica queens for hygienic behavior is beneficial for commercial beekeeping in Serbia, since hygienic behavior is associated with the resistance to the mite Varroa destructor, as well as to American foulbrood and chalk brood. In this work, heritability of hygienic behavior was analyzed in autochthonous grey A. m. carnica honey bees from Šumadija region in order to ascertain if the expression of their hygienic behavior could be enhanced through selective breeding. Heritability of hygienic behavior was monitored through three queen generations and estimated by one-parent-offspring regression method (motherdaughter regression method). The heritability values for hygienic behavior were h2=0.63±0.02 in relationship between daughter queens (F1s) and breeder mother queens (Ps), h2=0.45±0.01 in relationship between grand-daughter queens (F2s) and Ps and h2=0.44±0.02 in relationship between F2s and F1s. These results show that the expression of hygienic behavior through selective breeding could be enhanced and the best results could be achieved in the F1 generation. It is important to emphasize that the F1 colonies bred for hygienic behavior in our study suffer no apparent costs in reproductive and productive performances.Odgajanje matica sive medonosne pčele A. m. carnica sa ispoljenim higijenskim ponašanjem od velikog je značaja za komercijalno pčelarenje u Srbiji, obzirom da je higijensko ponašanje povezano sa otpornošću prema krpelju Varroa destructor, kao i prema izazivačima bolesti američke truleži pčelinjeg legla i krečnog legla. U ovom radu, ispitivana je heritabilnost higijenskog ponašanja kod autohtone sive medonosne pčele A. m. carnica iz Šumadije da bi se utvrdilo da li ispoljenost higijenskog ponašanja kod tih pčela može da se poveća putem selektivnog gajenja. Heritabilnost higijenskog ponašanja praćena je kroz tri generacije matica i procenjivana metodom regresije jedan roditelj-potomak (metoda regresije majka-ćerka). Vrednosti koeficijenta heritabilnosti higijenskog ponašanja bile su h2=0,63±0,02 između ćerki matica (F1s) i majki matica (Ps), h2=0,45±0,01 između unuka matica (F2s) i Ps, i h2=0.44±0.02 između F2s i F1s. Ovi rezultati pokazuju da ispoljenost higijenskog ponašanja kod analiziranih pčela može biti povećana, kao i da se najbolji rezultati mogu postići u F1 generaciji. Pri tome je značajno istaći da kod društava F1 generacije odgajenih na higijensko ponašanje u našem radu nije bilo nikakvih negativnih posledica u pogledu reproduktivnih i produktivnih sposobnosti

Učestalost hromozomskih aberacija u funkciji evaluacije genotoksičnog potencijala fumagilina in vivo

Dicyclohexylamine (fumagillin), as an antibiotic produced by fermentation of Aspergillus fumigatus, is used in human medicine for the treatment of intestinal microsporidiosis in patients with HIV infection, intestinal amebiasis and microsporidial keratoconjunctivitis. In veterinary medicine fumagillin is effective in suppressing microsporidiosis of bees and fish. In this study, the genotoxicity of fumagillin was evaluated in mouse bone marrow cells using chromosome aberrations (CA) assay. Dicyclohexylamine was administered to mice by gavage in a dose of 25, 50, 75 mg/kg b.w., with water-sugar syrup as the negative control and cyclophosphamide as the positive control (40 mg/kg b.w) Significantly increased frequency (p lt 0.01 or p lt 0.001) of numerical chromosomal aberrations (aneupliodies and polyploidies) and structural chromosomal aberrations of gaps, breaks and centric rings were observed only at the highest experimental dose of dicyclohexylamine, compared with the negative control. However, in point of induction of Rb translocations, both the median (50 mg/kg b.w) and highest (75 mg/kg b.w) experimental dose showed a significant (p lt 00.001) increase (7.12 ± 0.26 and 9.00 ± 0.10, respectively) in comparison with the negative control (0.00 ± 0.00). Chromosomes 4 and 19 participated in these Rb translocations. These results suggest that dicyclohexilamine (fumagillin) has genotoxic potential in mammal in vivo chromosomal aberration (CA) test system.Dicyclohexylamin (fumagilin), kao antibiotik koga proizvodi gljivica Aspergillus fumigatus, se koristi u humanoj medicini u borbi protiv intestinalnih mikrosporidioza kod pacijenata sa HIV infekcijom, ali i za tretman obolelih od intestinalnih amebijaza i mikrosporidijanog keratokonjunktivitisa. U veterinarskoj medicini efikasan je u lečenju mikrosporidioza pčela i drugih insekata uzrokovanih sa Nosema apis, N. cerani, N. bombici, ali i mikrosporidioza riba uzrokovanih sa Myxobolus cerebralis i Tetracapsuloides bryosalmonae. U ovom radu ispitivana je genotoksičnost dicikloheksilamina kvantifikacijom i kvalifikacijom hromozomskih aberacija u ćelijama kostne srži miševa soja BALB/c. Dicikloheksilamin je aplikovan miševima intragastrično u dozama od 25, 50, 75 mg/kg b.w., pri čemu je vodeno-šećerni sirup predstavljao negativnu, a cyclophosphamid (40 mg/kg b.w) poozitivnu kontrolu. Utvrđeno je da samo najveća doza dicikloheksilamina signifikantno povećava učestalost (p lt 0.01 ili p lt 0.001) numeričkih hromozomskih aberacija (aneupliodija i poliploidija) i strukturnih hromozomskih aberacija tipa gapova, prekida i centričnih ringova u poređenju sa negativnom kontrolom. Međutim, srednja (50 mg/kg b.w) i najviša (75 mg/kg b.w) eksperimentalna doza izazvale su signifikantno (p lt 0.001) povećanje (7.12 ± 0.26 i 9.00 ± 0.10, respektivno) Robetrsonovih (Rb) translokacija u poređenju sa negativnom kontrolom (0.00 ± 0.00). U formiranju ovih Rb translokacija utvrđeno je da učestvuju hromozomi 4 i 19. Ovi rezultati sugerišu da je dicikloheksilamin (fumagilin) ispoljio visok genotoksični potencijal sa sposobnošću indukcije i Rb. Translokacija

Heritabilnost negovateljskog ponašanja sive medonosne pčele (Apis mellifera Carnica)

Grooming behavior is considered an important defensive mechanism of honey bees against Varroa mites. The aim of this study was to reveal whether grooming behavior is a useful criterion in breeding of Varroa-tolerant bees. To obtain a reliable evaluation the environmental influences were excluded. The degree of grooming potential was estimated by the percentage of damaged mites in the total number of fallen mites. The heritability of grooming behavior throughout the three consecutive generations of queens was assessed by mother-daughter regression method. Among unselected queens, expressed grooming behavior was recorded only in colonies with F1 queens (36.27%), but not in colonies with P queens and F2 queens (33.69%, 31.66%, respectively). Significant differences in grooming behavior were found between colonies of P and F1 queens (p lt 0.001), and between colonies of P and F2 queens (p lt 0.05). However, all of the three generations of selected queens showed expressed grooming behavior (37.99%, 39.42% and 38.58% in Ps, F1s and F2s, respectively) without significant (p>0.05) difference among them. Nevertheless, the relatively low heritability of grooming behavior in the three generations of queens examined (h2yx=0.49±0.02; h2zx=0.18±0.01; h2zy=0.16±0.01) indicate that breeding colonies for grooming behavior only cannot be advised to beekeepers whose aim is to breed bees highly tolerant to Varroa mites.Negovateljsko ponašanje se smatra značajnim mehanizmom odbrane pčela od Varroa krpelja. Cilj ovog rada je bilo ispitivanje negovateljskog ponašanja, procena njegove heritabilnosti i mogućnosti povećanja ekspresije te osobine putem selekcije. Radi dobijanja pouzdanih rezultata korišćenja je metodologija kojom se uticaj spoljašnjih faktora isključuje. Ispoljenost negovateljskog ponašanja procenjivana je na osnovu procenta oštećenih u ukupnom broju otpalih krpelja. Heritabilnost negovateljskog ponašanja praćena na kroz generacije matica i procenjivana metodom regresije majka-ćerka. Među neselekcionisanim maticama, negovateljsko ponašanje bilo je izraženo samo kod matica F1 generacije (36,27%), ali ne i kod P (33,69%) i F2 generacije (31,66%). Statistički značajne razlike u negovateljskom ponašanju zabeležene su između društava P i F1 matica (p lt 0,001) i između društava P i F2 matica (p lt 0,05). Međutim, selekcionisane matice sve tri generacije (Ps, F1s, F2s) su imale izraženo negovateljsko ponašanje (37,99%, 39,42% i 38,58%) bez statistički značajnih (p>0,05) razlika među njima. Ipak, nizak koeficijent heritabilnosti praćene osobine (h2yx=0,49±0,02; h2zx=0,18±0,01; h2zy=0,16±0,01) ukazuje da se pčelarima ne može preporučiti selekcija pčela samo na negovateljsko ponašanje ako je njihov cilj uzgoj pčelinjih zajednica povećane otpornosti na Varroa krpelje

Evaluation of 11 microsatellite loci for their use in paternity testing in Yugoslav Pied cattle (YU Simmental cattle)

Eleven microsatellite loci (TGLA227.BM2113, TGLA53, ETH10, SPS115, TGLA126, TGLA122, INRA023, ETH3, ETH225, BM1824) were evaluated for their use in paternity testing in the Yugoslav Pied cattle (YU Simmental cattle) population in Serbia. A total of 40 animals were tested. At the 11 tested loci, a total of 91 alleles were detected. The mean number of alleles per locus was 8.273. Polymorphism information content (PIC) values ranged from 0.58 to 0.88 with the mean value of 0.72. The most informative loci were: TGLAS3 (14 alleles, PIC = 0.88), TGLA227(11 alleles, PIC = 0.82), INRA023 (11 alleles, PIC = 0.86), BM2113 (9 alleles, PIC = 0.80). Combined power of discrimination (CPD) for the 11 microsatellite loci was 0.999. The results of the present study confirm that the analysed set of 11 microsatellite markers recommended by ISAG is suitable for paternity testing in Yugoslav Pied cattle in Serbia

Genetic variation of apis mellifera from Serbia inferred from mitochondrial analysis

Two honeybee subspecies inhabit Serbia; Apis mellifera carnica and A. m. macedonica. Both belong to eastern Mediterranean (C) evolutionary lineage. Furthermore three Serbian honeybee ecotypes restricted to particular regions, were defined through morphometry and cytogenetic analyses. In this study, mitochondrial data have been used to analyze the molecular diversity of the honeybee population from Serbia. Seven haplotypes of the C evolutionary lineage have been found, two of them are newly described (C2o and C2p) and restricted to two regions, which ultimately increased the number of haplotypes found in this lineage. Comparisons with surrounding honeybee populations suggest a hybrid situation between A. m. carnica and A. m. macedonica and also introgression from A. m. ligustica. The results should be considered when dealing with future conservation strategies, and for pathogen-parasite-tolerant breeding programs

Banat donkey, a neglected donkey breed from the central Balkans (Serbia)

The dominant donkey breed in the Balkans is the mid-sized Balkan donkey with a grey to chocolate coat color. Local breeders from Serbia, however, still maintain a few larger individuals of a lighter coat color, named Banat donkey, and speculate that they are descendants of a Spanish donkey heard that had been transferred to the Banat region by the Hapsburg Queen Maria Theresa in the XVIII century for a specific purpose, to work in local vineyards. We have previously found a unique nuclear gene-pool and a prevalence of mitochondrial Clade 2 haplotypes in several such animals. In this study, we: (i) perform a comparative analysis of 18 morphological traits of the Banat donkey (seven individuals), Balkan donkey (53 individuals from two sub-populations of this breed) and the potential hybrids (eight individuals), and demonstrate the morphological distinctiveness of the Banat donkey, highlighting the diagnostic traits for distinguishing the breed: hip height, croup width, body length and chest depth; (ii) reanalyse published nuclear microsatellite data for these groups, and reveal that, although severely depopulated, the genetically distinct Banat donkey is not severely affected by the loss of genetic diversity and inbreeding; (iii) demonstrate that previously published Banat donkey mitochondrial haplotypes, analyzed genealogically together with those reported in ancient and modern individuals from Spain, Italy, Turkey, Cyprus and Africa, are shared with three Spanish breeds and individuals belonging to Amiata and some other Italian breeds. A unique morphological feature present in Banat and Somali wild donkeys, but also in Amiata donkeys, black stripes on legs, suggests that the origin of Clade 2 donkeys may be much more complex than previously thought. Actions to preserve the Banat donkey, a valuable but critically endangered genetic resource (<100 individuals), are urgent

Dietary conjugated linoleic acid influences the content of stearinic acid in porcine adipose tissue

The present study was conducted in order to determine the effects of supplementation of a growing-finishing pig diet with 0.5% conjugated linoleic acid (CLA) on production characteristics and slaughter traits. Ninety-seven female Swedish Landrace pigs were used. The control group of animals was fed a regular diet (n = 49), while the experimental group of animals (n = 48) received a diet where part of the soybean oil was substituted with commercially enriched CLA oil (containing at least 56% of CLA isomers, 28% cis-9, trans-11 and 28% trans-10, cis-12). The experiment lasted 44 days; porkers were fed from an initial weight of 66.0 up until a final weight of 103.5 kg. Feed conversion ratio, carcass and ham weight, percentage of lean meat and subcutaneous fat tissue as well as intramuscular fat were recorded. The fatty acid content of ham intramuscular fat tissue was determined by HPLC. No statistically significant influence of CLA was observed, either on carcass and ham weight, or on fat percentage in subcutaneous and intramuscular tissue. Dietary CLA enrichment proved to increase the content of stearinic acid in intramuscular fat tissue, 17.29 13.26 % in experimental and 15.87 33.71 % in control group of pigs (P < 0.01). The obtained production results show no statistically significant changes in main production traits between the two groups of animals. The observed difference in the content of stearinic acid (P < 0.01) implies firmer fat tissue, which has a practical value in pig bacon fattening

Uticaj cimiazol hidrohlorida na mitotsku i proliferativnu aktivnost u kulturama humanih limfocita

Cymiazole hydrochloride is the active ingredient of a synthetic systemically acting acaricide used in beekeeping (Apitol®, Apichem®). Considering that residues of cymiazole hydrochloride were detected in all bee products being used for human nutrition and as alternative medications in human and veterinary medicine, the objective of this study was to evaluate the effects of cymiazole hydrochloride on mitotic activity of cultured human lymphocytes. The mitotic and proliferation indices in phytohaemagglutinin-activated human peripheral blood lymphocytes were determined using three experimental concentrations of cymiazole hydrochloride (0.01; 0.1; 1mg/ml). All experimental concentrations of cymiazole hydrochloride increased mitotic index values (MI) by a statistically highly significant amount (p lt 0.001). Moreover, proliferation index (PI) was also changed very significantly (p lt 0.001) after each experimental cymiazole hydrochloride treatment. The dose dependent increase in proliferation index values clearly demonstrates the ability of cymiazole hydrochloride to induce significant alterations of cell cycle kinetics having an influence on G0 state and arousing cell division. This indicates a risk from consuming bee products with cymiazole hydrochloride residues particularly in those predisposed to malignant diseases. Strict respect to the withrawal time while using cymiazole hydrochloride (Apitol®, Apichem®) is of great importance because residue levels higher than the permitted one can easily occur in honey and other bee products from treated honey bee colonies.Cimiazol hidrohlorid predstavlja aktivnu supstancu sintetičkih akaricida sa sistemskim dejstvom koji se koriste u pčelarstvu (Apitol®, Apichem®). Obzirom da su rezidue cimiazol hidrohlorida detektovane u svim pčelinjim proizvodima, koji se koriste u ishrani i kao alternativni lekovi u humanoj i veterinarskoj medicini, cilj ovog rada bio je ispitivanje efekata cimiazol hidrohlorida na mitotsku i proliferativnu aktivnost humanih limfocita u kulturi. Analizirani su mitotski i proliferativni indeksi humanih limfocita aktiviranih fitohemaglutininom pri čemu su koršćene tri eksperimentalne koncentracije cimiazol hidrohlorida u opsegu od 0.001 mg/ml do 1 mg/ml. Sve tri eksperimentalne koncentracije cimiazol hidrohlorida izazvale su vrlo visoko signifikantno (p lt 0.001) povećanje vrednosti mitotskog indeksa (MI) humanih limfocita. Takođe, vrednosti proliferativnog indeksa (PI) su vrlo visoko signifikantno (p lt 0.001) povećane nakon svakog eksperimentalnog tretmana cimiazol hidrohloridom. Dozno zavisno povećanje vrednosti proliferativnog indeksa jasno pokazuje sposobnost cimiazol hidrohlorida da izazove značajne promene u kinetici ćelijskog ciklusa utičući na G0 tačku i podstičući ćelijsku deobu, što ukazuje na rizik prilikom konzumiranja pčelinjih proizvoda sa reziduama cimiazol hidrohlorida, posebno kod jedinki sklonih malignitetima. Izuzetno je značajno strogo poštovanje karence prilikom upotrebe cimiazol hidrohlorida (Apitol®, Apichem®) jer rezidue, u količini većoj od dozvoljene, mogu veoma lako dospeti u med i druge pčelinje proizvode dobijene od tretiranih pčelinjih društava

A molecular genetic approach to roebuck individual identification in the case of poaching in Serbia

Application of the molecular genetic methods in forensic cases dealing with wild animals has significantly increased recently. These techniques are practically used in order to help solving four key problems : determination of kind of the wild animal, geographic origin, kinship ties and individual identification. In this work the first case of introducing the examination of polimorphism of microsatelite genetic markers within forensic analysis in the cases of poaching in Serbia is presented. The objectives of this forensic analysis was to determine if the meat confiscated during house search of the suspect comes from roebuck origin (Capreolus capreolus), which remains had been found by a game warden in the field during closed season, where the suspect denied the offense, claiming that the meat comes from other roebuck that had been shot during the previous hunting season. DNK was isolated from the skin and fur samples taken from the roebuck corpse found in the woods, as well as from the frozen meat found in the suspect’s house. Both amplification and polimorphism examination of the eight microsatelite markers (ROE01, NVHRT21, NVHRT24, NVHRT48, NVHRT73, RT7 AND RT27) were carried out. In all the examined samples, the same pattern of variability of the tested microsatelites was determined, that is it was proved that DNK profiles of the samples taken from roebuck corpse were identical to DNK profile of the meat sample found in the suspect’s house. This result clearly indicates that all the examined biological samples originate from the same animal, and consequently represents forensically valid evidence in the case of roebuck poaching. [Projekat Ministarstva nauke Republike Srbije, br. III46002