proceedings of the international symposium on animal ...the first written record of thevarieties of...

Proceedings of the International Symposium on Animal Science 2016,

24-25th November 2016, Belgrade, Serbia

The publishing of this book was made possible by the support of Ministry of Education, Science and Technological Development and Ministry of Agriculture and Environmental protection of Republic of Serbia.

All papers of the International Symposium on Animal Science 2016 have been reviewed by the members of the International Scientific Committee and competent reviewers proposed by the International Scientific Committee.

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INTERNATIONAL Symposium On Animal Science (2016 ; Belgrade) Proceedings of the International Symposium on Animal Science 2016,24-25November 2016, Belgrade, Serbia [Elektronski izvor] / [editor in chiefZoran Popović]. - Belgrade : University, Faculty of Agriculture, 2016(Belgrade : Pekograf). - 1 elektronski optički disk (CD-ROM) ; 12 cm

Sistemski zahtevi: Nisu navedeni. - Nasl. sa naslovne strane dokumenta.-Tiraž 150. - Bibliografija uz svaki rad.

ISBN 978-86-7834-261-5

a) Сточарство - ЗборнициCOBISS.SR-ID 227407884

University of Belgrade,Faculty of Agriculture, Nemanjina 6, 11080 Belgrade-Zemun, Belgrade

Prof. Dr Milica Petrovic Prof. Dr Zoran Popovic Vladimir BrašanacFaculty of Agriculture, Nemanjina 6, 11080 Zemun, Belgrade Pekograf d.o.o Vojni put 2, 11080 Belgrade

150

II

INTERNATIONAL SYMPOSIUM ON ANIMAL SCIENCE 2016 24-25 November 2016, Belgrade-Zemun, Serbia

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Poster presentation

FOREWING DIFFERENCES BETWEEN HONEY BEES FROM BANAT AND CENTRAL SERBIA

Nedić N.1, Mirjanić G.2, Jevtić G.3, Anđelković B.1, Plavša N.4

Abstract

The first written record of the varieties of honey bees in Serbia may be found in the work of Grozdanić (1926) who performed morphological comparisons of honey bee samples from Banat with A. m. carnica, A. m. ligustica, A. m. cypria and A. m. syriaca and suggested the separation of the bees from Banat into subspecies A. m. banatica. Due to the adaptable abilities of bees to specific habitat conditions in the Pannonian plain, it is assumed that different geographic ecotypes were created there.

Because of this the aim of our research was to analyze a part of morphological characters of honey bees from Banat and to compare them with honey bees from Central Serbia, in order to help distinguish and preserve indigenous honey bee varieties.

The samples were collected in three different locations in Banat and one location in Central Serbia. In this study, 14 morphometric characters were measured in accordance with the standard method. By means of a variance analysis very significant differences were determined between the two examined bee groups. On the basis of LSD test we have determined that some honey bee groups from Banat differed very significantly (P<0.01) from honey bees in Central Serbia regarding A4, D7 and E9 angles. Honey bees from Uljma (Southern Banat region) differed very significantly from examined honey bees as regards the size of O26 angle (42.11±3.03o). Group of bees from Central Serbia had a very significantly larger width of forewing in relation to Banat honey bees. Investigation of morphometric characters on the forewing could contribute to a clearer separation of groups of bees inside domestic population.

Keywords: honey bee, morphometry, Banat, Central Serbia

1Nedić Nebojša, PhD, Associate Professor; Anđelković Bojan, PhD student, University of Belgrade, Faculty of Agriculture, Belgrade, Serbia; 2 Mirjanić Goran, PhD, Assistant Professor; University of Banja Luka, Faculty of Agriculture, Banja Luka, Bosnia and Herzegovina; 3 Jevtić Goran, PhD, Research associate, Institute for Forage Crops, Globoder, Kruševac, Serbia; 4 Plavša Nada, PhD, Associate Professor, University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia; Corresponding author: Nedić Nebojša, email: [email protected]

mailto:[email protected]


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Introduction

Apis mellifera L. is an indigenous species in Europe, Africa, and Asia (including Saudi Arabia, Iran, and the Ural Mountains in Russia). This species is widespread all over the world due to multiple migrations and introductions by humans (Meixner et al., 2013). Apis mellifera has about 29 subspecies in different geographic regions of the world and there are significant differences between them (Munoz et al., 2009).

Honey bees adapt to different environmental conditions in which they live. In order to achieve excellent production results, for which the beekeepers are most interested, breeders select lines with desirable characteristics (Nedić et al., 2011a). Selection of more productive honey bees with uniform performance can lead to uniformity of bee populations thus reducing genetic variability and adaptation to local conditions (Meixner et al., 2013).

Morphometry is a good method used in taxonomy and systematics of bees (Stevanović, 2002). Set of 36 morphometric characteristics used by Ruttner (1988) was utilized for the most famous biographical honey bee taxonomy and establishment of "classical" morphometry (Čapek and Chlebo, 2016). DuPraw (1965) introduced the measurement of 11 angles on the front wing, which proved to be very useful in research, because these characters are less influenced by environmental conditions and, thus, they represent good indicators of genetic relationships between populations (Meixner et. al. 2007; Abou- Shaara, 2013).

The first written record of the varieties of honey bees in Serbia can be found in the paper published by Živanović (1893) from the end of XIX century. He states that, in Srem, there are two varieties of honey bees, one dark and other resembles the Italian bee (Apis mellifera ligustica) because of two yellow rings on the abdomen.

Grozdanić (1926) suggested the separation of the bees from Banat into subspecies A. m. banatica based on morphological comparisons of samples of honey bee from Banat with A. m. carnica, A. m. ligustica, A. m. cypria and A. m. syriaca. Krunić (1967), studying the variability of honey bees from the area of the Pannonian Plain, concluded that they constitute a homogeneous whole, which should be distinguished as a race A. m. panonica. In her later studies, Stevanović (2002) researched honey bees of Sjenica-Pester, Timok and Banat ecotypes. The largest morphological differences were found between the Sjenica- Pester and Banat honey bee ecotypes. According to Mladenović et al. (2011), there is variability in the morphological characters of yellow bees in northern Serbia. Pihler (2012) notes that, in the geographical area of Vojvodina, honey bee populations are reminiscent to the races Apis mellifera carnica, Apis mellifera macedonica, Apis mellifera ligustica, considering their morphometric characteristics.

It is important to identify variation within Serbian honey bee population, in order to help distinguish and preserve indigenous Carniolan honeybee. The aim of this study was to analyze the size of the 14 morphometric traits of honey bee samples from Banat and from Kraljevo (central part of Serbia).


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Material and Methods

Samples of 15 adult worker bees (Meixner et al., 2007 and 2013) were collected from 4 different apiaries along Serbia: Gudurica (45o17´07.22´Ń, 21o44´80.25´É) (group I), Vršac (45o13´19.17´Ń, 21o35´51.06´É) (group II), Uljma (45o05´05.81´Ń, 21o16´06.52´É) (group III), Kraljevo (43o70´00.50"N 20o60´51.44"E) (group IV). Bees were preserved in 95% ethanol, and then dissected, and the right forewing were mounted on glass slides and measured using Leica XTL-3400D binocular microscope and software package IL 1009 in accordance with the standard method.

The following characters were used for measurements: angle A4, angle B4, angle D7, angle E9, angle L13, angle J10, angle J16, angle N23, angle K19, angle G18, angle O26, cubital index (CUBI), forewing length (FWL); forewing width (FWW).

Univariate (variance) statistical analyses were conducted for 14 morphological traits of worker bee samples from different locations of the territory of Serbia. A descriptive statistical analysis was carried out and comparisons between locations were determined by LSD Test. All measurements of morphological characteristics for the bees were analyzed by multivariate discriminant analyses.


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Results and Discussion

The values of descriptive statistics of morphological characteristics of honey bees from the site of Banat and Royal are shown in Table 1. Table 1. Means and standard deviation (Sd) of selected measures for the bee samples. Sizes of angles are given in degree (o), size of other morphometrical characters are given in micrometer (µm). Char.

Origin of Samples (locations) Average Gudurica (I) Vršac (II) Uljma (III) Kraljevo (IV)

A4 29.96±1.72 ab*

28.91±1.77 a

30.53±2.42 ab

31.22±1.58 b

30.15±2.00

B4 109.77±3.99 a

108.34±5.44 a

108.94±5.41 a

104.70±4.21 a

107.94±5.08

D7 97.83±2.33 b

95.93±3.13 ab

97.13±2.93 b

93.44±2.79 a

96.08±3.22

E9 23.70±1.55 b

22.38±1.44 ab

22.99±1.33 b

21.44±1.46 a

22.63±1.64

L13 14.25±1.25 a

15.03±1.17 a

13.82±1.90 a

14.69±2.04 a

14.49±1.43

J10 56.09±3.29 a

51.22±2.36 a

52.73±2.66 a

52.77±3.88 a

53.67±3.56

J16 90.01±2.03 a

93.45±2.96 a

88.65±3.58 a

92.72±2.10 a

91.44±3.56

N23 94.87±3.19 a

93.61±2.63 a

90.26±2.40 a

91.03±2.23 a

92.57±3.62

K19 77.01±1.44 a

78.18±2.18 a

79.61±2.61 a

79.19±2.07 a

78.36±2.45

G18 91.50±2.06 a

91.26±3.94 a

90.13±4.21 a

89.16±3.51 a

90.69±3.10

O26 37.32±2.82 a

35.39±3.01 a

42.11±3.03 b

37.07±4.33 a

37.97±4.13

FWL 9061.93±109.0 2

8841.90±97.1 1

8926.35±155.5 6

9297.13±123.4 3

9031.83±211.0 4

FW W

3207.13±54.74 a

3143.88±91.3 7

3152.25±76.47 a

3320.37±69.01 b

3205.91±101.2 4

CUBI 2.54±0.36 a

2.52±0.42 a

2.45±0.44 a

2.69±0.37 a

2.55±0.40

*Means for the same characteristics followed by different letters within locations are significantly different (P<0.01) according to variance analysis followed by LSD test.

The average value of the angle A4 varied from 28.91° (group II-Vršac) to 31.22° (group IV-Kraljevo). The present results are in agreement with the results of Nedić et al. (2011b). Pihler (2012), in his research, shows slightly higher value of angle A4 from 29.74° ± 4.85° in bees from Banat. Analysis of variance showed that there is a statistically significant difference (P <0.01) between the angles A4 in group II (Vršac) and group IV (Kraljevo). The size of angle B4 in the observed samples ranged from 104.70° (group IV) to 109.77° (group I), with an average value of 107.94°. In his studies, Cermak (1999) states that the


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honey bees of Carniolan race had the value for B4 angle of 107.4°. Somewhat different values are stated by Stevanović (2002) for bees of Sjenica-Pešter, Banat and Timok ecotype, that had an average size of the angle B4 109.3°, 111.2° and 109.9°, respectively.

Variation interval of the angle D7 in examined groups of bees ranged from 93.44° to 97.83° with an average value of 96.08°. These results are consistent with the results Stevanović (2002) in which the size of the angle D7 ranged from 94.9° to 96.7°. Kauhausen-Keller and Keller (1994) found a slightly higher value for the test character (97.5°), in their morphological research of bees from lines of Apis mellifera carnica from beekeeping institute in Kirchhain. Analysis of variance showed that there is a very significant difference (P <0.01) in the size of the angle D7 between the groups of honey bees from central Serbia and honey bees from the area of Banat (groups I and III).

In the observed groups of bees the size of the angle E9 was the lowest in the group IV (21.44°), and the largest in the group I (23.70°), with an average of 22.63°. The results are consistent with the results Ruttner (1988) who stated value of 12.23 ± 2.08° for the size of the angle E9 for A. m. carnica. Slightly lesser values for angle E9 in bees of Sjenica-Pešter, Banat and Timok ecotype (18.9°, 20.0° and 19.7°, respectively) were found in studies conducted by Stevanović (2002). Results of the analysis of variance showed a significant difference between the groups of bees from central Serbia and bees from all three groups from the area of Banat.

Average values for angles L13, J10, J16, N23, K19 and G18 were 14.49±1.43°, 53.67±3.56°, 91.44±3.56°, 92.57±3.62°, 78.36±2.45° and 90.69±3.10°, respectively. Determined differences of average values of these angles between the groups of bees were not statistically significant.

The angle O26 in the observed samples ranged from 35.39° (group II) to 42.11° (group III), with an average value of 37.97°. The results for this characteristic are in accordance with the values for Carniolan race. According to investigation of Maul and Hahn (1994), the size of angle O26 in the reference sample of A. m. carnica was 37.97 ± 3.6°. Pihler (2012), in the study of bees from Banat, found the average value of the angle E9 (39.88 ± 5.52°). Analysis of variance showed that the values of these angles differ significantly between sites (P <0.01).

Beside certain angles, the discrimination of groups of bees was very significantly influenced by the other studied morphometric characters on the wings. The data obtained by descriptive statistics (table 1) show that there are variations from 8841.90 μm (group II) to 9297.13 μm (group IV) in the length of the front wing. Bees from Central Serbia (IV) differed significantly for this character (P <0.01) from groups I, II, III and VI of bees from Banat. Also, a very significant difference (P <0.01) in the width of the front wing was found between group of bees from Central Serbia (IV) and groups of bees originated from Banat. Wing width ranged from the 3143.88 μm (II) to 3320.37 μm in the observed samples, with an average value of 3205.91 μm. A similar value of this character (3.23mm) for the bees from Banat was reported by Jevtić (2007).

Value of cubital index in our research ranged from 2.45 (III) to 2.69 (IV), with an average value of 2.55. The values were in the range for Carniolan race. Ruttner (1988) stated that


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the value of the cubital index for A. m. carnica is 2.589±0.418. No statistically significant difference between the groups of bees was determined for cubital index.

For the characterization of subpopulations within races, multivariate statistical methods may be used to estimate similarities and/or dissimilarities (Kauhausen-Keller and Keller, 1994).

Graph 1. View of the distribution of bee groups centroids by location (CV1 vs. CV2)

The results of multivariate discriminant analyses of the measurements of the 14 characteristics showed significant differences existed between the four groups of bees (Wilk´s λ = 0.0297, P<0.00001). The 1st axis (CV 1) explained 79.7%, the 2nd (CV 2) 15.4% and the 3rd axis (CV 3) 4.9% of the total variation. Graph 1 shows the distribution of the centroids of the analyzed bee samples according to the first discriminant axis which explains most of the total variability. According to these analyses the local honey bee samples from Kraljevo in Central Serbia (group IV) showed most difference compared to most other groups of bees originated from Banat (Graph 1).

The group of bees from Vršac was the most different from the other two groups sampled bees from Banat (Graph 1). Distinguishing between groups of honey bees is contributed mostly to morphometric characters: angle A4, angle D7, angle E9, angle O26, length and width of the front wings.

Based on the results of this analysis, we could speak about the honey bee population from Banat that are similar to each other and their common difference from the pattern of bees from central Serbia. Various geographical, climatic and botanical conditions in areas have caused a differentiation of Carniolan bee subspecies. Then there is the function of anthropogenic factors, which is reflected in the unplanned replacement of honey bee queens of unknown origin and production capabilities.

Root 1 vs. Root 2 6

5

4

3

2

1

0

-1 -2 -3 -4

-6 -4 -2 0

Root 1

2 4

I II

6 III IV

Root

2


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Conclusion

Based on the results obtained by morphometric analyses of 14 characteristics of the front wings of bees from the site of Banat and Central Serbia it can be concluded that there is a great phenotypic variability. The analysis of variance showed that the sampled groups of bees highly statistically significantly differed in 6 properties (A4, D7, E9, O26, FWL, FWW).

Very significant differences (P <0.01) between the bees from the area of Banat and groups of bees from central Serbia occurs in 35.7% of the observed characteristics. Very significant differences (P <0.01) between groups of bees from the area of Banat occurs only in 14.3% of the observed characteristics.

Results of the multivariate discriminant analysis show significant differences between the sampled group of bees (Wilk's λ = 0.0297, P <0.00001), where a group of bees from Central Serbia was clearly distinguished in relation to the sampled bees from Banat.

The values of certain morphometric characters of examined especially to the group of bees from Kraljevo showed a deviation from the standard values for Carniolan honey bee race. Variability in honey bee phenotype determined for groups of bees from Banat indicate the existence of a population of bees that mutually differ less in comparison with bees from central Serbia. Bees from the Banat area were sampled in the plain area, while the bees from central Serbia were sampled from the area that can be considered mountainous. Various environmental factors like relief and flora influenced the adjustment of bees to different conditions of life prevailing in these two areas, and are able to influence the inter- variability examined groups of bees. We should also not rule out the influence of man through migratory beekeeping, free replacement of honey bee queens of unknown origin and with it the possibility of an impact on the existing bee populations in Serbia.

Research of characters of the front wings has contributed to the separation of the sampled group of bees. Determined variability provides an opportunity for further promotion and selection of bees that meet the criteria for the morphometric traits established for the Apis mellifera carnica.

Acknowledgment

The research was financed by the Ministry of Science and Technological Development, the Republic of Serbia projects 46008 and 46009. We are grateful to all beekeepers that contributed to worker bee samples.


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References

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http://www.beedol.cz/dawino/DAWINO_protocol_EN.pdf

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