V-XVII a. Lietuvos arklių plaštakų ir pėdų osteometrinė analizė bei biologinio amžiaus nustatymas pagal dantų struktūrą ; Osteometrical analysis of metacarpal and metatarsal bones of V-XVII c. Lithuanian horse and biological age determination according to teeth structure

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LITHUANIAN VETERINARY ACADEMY The research work has been carried out at the Lithuanian Veterinary Acad- emy, Department of Anatomy and Physiology, in 2002–2006. Research supervisor – Prof. at Incumbent dr. Linas Daugnora (Biomedical Sciences, Veterinary Medicine – 12 B), Lithuanian Veterinary Academy. Chairman of the Veterinary Medicine Council – Doc. dr. Albina Aniulien ė (Biomedical Sciences, Veterinary Medicine – 12 B), Lithuanian Veterinary Academy. Snieguol ė Veli čkait ė Members: Prof. dr. Bronius Bakutis (Biomedical Sciences, Veterinary Medicine – 12 B), Lithuanian Veterinary Academy; Prof. habil. dr. Vytautas Špakauskas (Biomedical Sciences, Veterinary Medicine – 12 B), Lithuanian Veterinary Academy; Prof. dr. Neringa Paužien ė (Biomedical Sciences, Medicine – 07 B), Kaunas University of Medical; OSTEOMETRICAL ANALYSIS OF Dr Laima Bal čiauskien ė (Biomedical Sciences, Ecology and Environmental Science – 03B), Institute of Ecology of Vilnius University. METACARPAL AND METATARSAL BONES OF V–XVII C. LITHUANIAN HORSE AND Opponents: BIOLOGICAL AGE DETERMINATION ACCORDING Dr. Ingrida Monkevi čien ė (Biomedical Sciences, Veterinary Medicine – TO TEETH STRUCTURE 12 B), Lithuanian Veterinary Academy; Doc. dr. Rimantas Jankauskas (Biomedical Sciences, Medicine – 07 B), Vilnius University.
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LITHUANIAN VETERINARY ACADEMY  
     Snieguol Veli č kait       OSTEOMETRICAL ANALYSIS OF METACARPAL AND METATARSAL BONES OF VXVII C. LITHUANIAN HORSE AND BIOLOGICAL AGE DETERMINATION ACCORDING TO TEETH STRUCTURE      SUMMARY OF DOCTORAL DISSERTATION BIOMEDICAL SCIENCES, VETERINARY MEDICINE (12B)       KAUNAS, 2006
 The research work has been carried out at the Lithuanian Veterinary Acad-emy, Department of Anatomy and Physiology, in 20022006.   Research supervisor Prof. at Incumbent dr. Linas Daugnora (Biomedical Sciences, Veterinary Medicine  12 B), Lithuanian Veterinary Academy.  Chairman of the Veterinary Medicine Council  Doc. dr. Albina Aniulien (Biomedical Sciences, Veterinary Medicine  12 B), Lithuanian Veterinary Academy.  Members: Prof. dr. Bronius Bakutis (Biomedical Sciences, Veterinary Medicine  12 B), Lithuanian Veterinary Academy; Prof. habil. dr. Vytautas pakauskas (Biomedical Sciences, Veterinary Medicine  12 B), Lithuanian Veterinary Academy; Prof. dr. Neringa Pauien (Biomedical Sciences, Medicine  07 B), Kaunas University of Medical; Dr Laima Bal č iauskien (Biomedical Sciences, Ecology and Environmental Science  03B), Institute of Ecology of Vilnius University.  Opponents: Dr. Ingrida Monkevi č ien (Biomedical Sciences, Veterinary Medicine  12 B), Lithuanian Veterinary Academy; Doc. dr. Rimantas Jankauskas (Biomedical Sciences, Medicine  07 B), Vilnius University.  Public defense of Doctoral thesis in Veterinary Medicine Science Council will take place at Lithuanian Veterinary Academy I auditorium 14 am on 30 th of November, 2006.   Address: Til s 18 LT-47181, Kaunas, Lithuania. The abstract of doctoral dissertation has been send on 30 th of October, 2006 according to the confirmed address list. This dissertation is available at the libraries of the Lithuanian Veterinary Academy and Veterinary Institute of Lithuanian Veterinary Academy.
LIETUVOS VETERINARIJOS AKADEMIJA            Snieguol Veli č kait       VXVII a. LIETUVOS ARKLI  PLATAK IR P D OSTEOMETRIN ANALIZ BEI BIOLOGINIO AMIAUS NUSTATYMAS PAGAL DANT STRUKT R Ą      Daktaro disertacijos santrauka Biomedicinos mokslai, veterinarin medicina (12 B)       Kaunas, 2006
Disertacija rengta 2002-2006 metais Lietuvos veterinarijos akademijos Anatomijos ir fiziologijos katedroje.  Mokslinis vadovas  E. prof. p. dr. Linas Daugnora (biomedicinos mokslai, veterinarin medicina  12 B), Lietuvos veterinarijos akademija.   Disertacija ginama Lietuvos veterinarijos akademijos Veterinarin s medicinos mokslo krypties taryboje:  Pirminink Doc. dr. Albina Aniulien  (biomedicinos mokslai, veterinarin  medicina    12 B), Lietuvos veterinarijos akademija.  Nariai: Prof. dr. Bronius Bakutis (biomedicinos mokslai, veterinarin  medicina  12 B), Lietuvos veterinarijos akademija; Prof. habil. dr. Vytautas pakauskas (biomedicinos mokslai, veterinarin  medicina  12 B), Lietuvos veterinarijos akademija; Prof. dr. Neringa Pauien (biomedicinos mokslai, medicina  07 B), Kauno medicinos universitetas; Dr. Laima Bal č iauskien  (biomedicinos mokslai, ekologija ir aplinkotyra  03B), Vilniaus universiteto Ekologijos institutas.  Oponentai: Dr. Ingrida Monkevi č ien , (biomedicinos mokslai, veterinarin  medicina  12 B), Lietuvos veterinarijos akademija; Doc. dr. Rimantas Jankauskas (biomedicinos mokslai, medicina  07 B), Vilniaus universitetas.   Disertacija bus ginama vieame Veterinarin s medicinos mokslo krypties tarybos pos dyje 2006 m. lapkri č io 30 d. 14 val. Lietuvos veterinarijos akademijos I auditorijoje. Adresas: Til s g. 18, 47181 Kaunas, Lietuva.   Disertacijos santrauka isiuntin ta 2006 m. spalio m n. 30 d. pagal pat-virtint ą  adres  s ą ra ą . Disertacij ą  galima peri r ti Lietuvos veterinarijos akademijos ir LVA Veterinarijos instituto bibliotekose.
INTRODUCTION  The first horse burials in Lithuania were found in the coastal burial ground dated to the 2d4th centuries (Volkait  -Kulikauskien , 2001; Michelbertas, 1984). Most abundant data were available from Central Lithuania where horse burials were known since the middle of the 1 st  cen-tury. Horse skeletons found in Marvel  burial ground (7th11th centuries) were most thoroughly investigated. Many publications were prepared based on this material (Daugnora, 1994; Daugnora, 1996; Bertaius, Daugnora, 1997; Bertaius, Daugnora, 2001). Horses remains uncovered in other places than burial grounds  hill forts, cities or castles have been less in-vestigated by osteologists so far. The objects mentioned above must be stud-ied for comprehensive and more objective knowledge and view about the types of horses bred in Lithuania and in the neighbouring countries (Poland, Russia, Latvia, etc.) in the past. Date of archaeological material from the 5th century included many well preserved horse bones what enabled following up the evolution of the Lithuanian horse until today. Material about horse height, constitution and age is very rare. Metacarpal and metatarsal bones, and skull or at least the lower jaw with teeth could be pointed out as most valuable material. Teeth belong to the best preserved elements of excavations. Such factors as dentition or wear allow determin-ing the animal age. However solitary molars (as they found the most fre-quently) do not able to provide this kind of information. Therefore, the methods allowing determining the horse age according to the tooth height and number of cementum rings are very.  The aim of the research The aim of the present work was to describe the whiter height, type and age of horses bred in the 5th17th centuries in the present territory of Lithuania and to specify and evaluate the methods of determining the bio-logical age of an individual.  The tasks of the research 1. Determining the wither height of horses bred in the 5th17th centuries in the territory of Lithuania. 2. Determining the types of horses according to the relative width of metacarpus and metatarsus bones. 3. Assessment of the level of objectiveness of the methods used for de-termining the biological age according teeth structure.  Originality of research:  Osteological material from the 5th17th centuries excavated in Lithuania was analysed and the whiter height and type of the then bred horses. The age
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of the excavated and present horses were determinate using the radiographic and computer tomography research methods and by counting rings of ce-mentum.  MATERIAL AND METHODS  The osteological and osteometric bone analysis was performed at the Laboratory of Osteology, Department of Anatomy and Physiology of the Lithuanian Veterinary Academy (LVA). The analysed osteological material was collected from the following ar-eas: Upper town of Kernav  (13th15th century); Pakalnikiai (12th15th century); Obeliai (12th14th century); Taurapilis (5th6th century); Skub -tai (9th12th century); Plinkaigalis (5th6th century); Degsn   Labotik s (5th12th century); Vilnius (14th  the first half of the 15th century and the second half of the 15th  17th century) and Marvel (11th17th century ). The skulls of present horses whose age were known were collected in 19962002. The horses were used for the purposes of students training. The used oseological material was stored at the repositories of the Laboratory of Osteology, Department of Anatomy and Physiology of the LVA.  Osteometry and determining of the horse height  The minimal number of individuals (MNI) was determined by T. Whites (1953) method. According to the relative width of diaphysis of metacarpal and metatarsal bones (SD*100/GL) the horses were classified into types within A. Brauners classification. The wither height of horses was determined using the V. O. Vitts (Vitt, 1952) methods. The metacarpal and metatarsal bones were measured by A. von den Driesch (1976) method. The bones were measured with the calliper (precision 0.1 mm).  Methods for determining the horse age  The horse age was determined according to the wear of incisors using Goodys (1997) schemes and corrected according to tables given by Muyllei and others (1999), M. Levine (1982), I. A. Silver (1969), A. F. Klimov (1955), R. Getty (1955).  Radiographic and computer tomographic images of teeth  The excavated material often contained individual molars or fragments of jaw without incisors what made more difficult to determine the horse age. Using the radiographic and tomogramphic images the teeth were measured and thus the horse age determined. The lower jaws of present horses of known age were taken and teeth measured to correct the radiographic and computer tomographic methods. The biological age of horses was determined based on the teeth height data.
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The height of teeth was statistically compared with the chronological age. The excavated horse teeth were also measured in radiographic imaging. The horse age determined by teeth measuring was compared with the horse age determined by other methods. The radiographic imagings were made at the LVA Dr L. Kriau č eli nas Clinics of Small Animals using Americomp spectra 325e apparatus (parameters of ray exposition 125 kV300 mA). The height of all molars between the masticator surface and furcation of the root and the length of teeth on the left side of the jaw were measured on the ra-diographic images. The computer tomograms were made at the Radiological Clinics of Kau-nas Medical University using 6-ply Siemens Spamatom Emotion and 16-ply General Electric Light speed multi-section computer tomographic scanners. The jaws were scanned by 2.5 mm thick layers. The height of all molars between the masticator surface and furcation of the root and the length of teeth on the left side of the lower jaw were measured on the computer tomo-graphy images. For correction of measuring data obtained from radiographic images and computer tomography, molars were chiselled from the lower jaw and their height, width and length were measured. The age of horses was determined according to the height of measured molars by M. Levine (1982) method.  Histological examination of teeth  Teeth sections were made at the Forensic Criminology Laboratory, Insti-tute of Forensic Medicine of Mykolas Romeris University. The first molars (M 1 ) from the lower jaw of present and excavated horses were taken for examination. Horizontal sections of the roots of horse teeth were made by methods described in the works of S. Trumpickait -Dzek č iorien (2000) and R. Bojarun (2003) and using microtome LEICA SP 1600. For determining the reliability of the data obtained histological prepara-tions of decalcinated teeth were made. These histological preparations were prepared by method described in C. Azorit et al. (2002).  Statistical data analysis  Statistical data analysis was carried out at the Department of Applied Mathematics of Kaunas Technological University using the SPSS 9.0 statis-tical analysis system for Windows. The following methods of statistical analysis were applied in the present work: 1. Descriptive statistics. Digital characteristics for analysed dimensions were calculated (frequencies, relative frequencies, minimal and maximal values, arithmetic mean and standard deviation (SD)). 2. Hypotheses about compatibility of actual and normal pattern of ana-
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lysed dimensions were checked using KolmogorovSmirnov criterion. The Wilcocksons criterion of signs was used for examination of dependent data sets. Statistical significance of the differences of metacarpal and metatarsal bones length in different localities and data on teeth height and length ob-tained from roentgenograms and tomograms was determined. 3. Correlation analysis. The strength of the link between the metatarsal bones length and width, chronological age of teeth and biological age de-termined by measuring teeth with the aid of trammel and in roentgenograms and tomograms, as well as between the number of cementum rings and chronological age was evaluated using Spidermans ranking correlation co-efficient. Hypotheses about significance of correlation coefficients (H o : cor-relation coefficient equals to zero) were checked. If zero hypothesis H o was rejected with significance value p<0.05 the correlation was considered sig-nificant. 4. Regression analysis was employed for assessment of relationship be-tween data sets. 5. The data were considered statistically significant when p<0.05.  RESULTS  Osteometry and whiter height of horse The length of metacarpal bones ranged from 170.2 mm to 250 mm (Ta-ble 1) and the length of metatarsal bones varied from 198.9 mm to 291 mm (Table 2).  Table 1. Length of the horse metacarpal bones  Locality Nofu bmobnesr  Avvaelruaeg e dSetavinadtiaordn Min Max Kernav  13 205,70 18,20 179,6 230 Vilnius 14ththe1st half of the 15th c. 5 207,1 17,5 188,0 229,9 tVhiel n1i5utsh th1e 72thd  chalf of 8 225,6 17,8 200,0 250,0 . Obeliai 2 180,45 14,07 170,5 190,4 Taurapilis 2 205,4 6,65 200,7 210,1 Plinkaigalis 2 210,65 0,35 210,4 210,9 Pakalnikiai 12 199,25 16,37 180,5 220,4 Degsn Labotik s 3 190,3 20,05 170,2 210,3 Marvel  72 192,8 8,35 181 216
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Table 2. Length of horse metatarsal bones y Number Max Localitof bones Average dSetavinadtiarodn Min Kernav  12 242 18,18 198,9 271,8 Vilniufs  t1he4 t1h5tthh ec1. st 3 241,3 15,9225,1 256,8 half o Vilnius the 2d half of the 15th17th c. 8 258,8 22,9 226,8 291,0 Obeliai 2 220 14 210,2 230 Taurapilis 2 240,7 0,28 240,4 240,8   Plinkaigalis 1 260,2 - - -Pakalnikiai 3 264,0 5,86 260,5 270,8 Degsn Labotik s 2 235,5 7,64 230,1 240,9 Marvel  48 234,14 8,83 219 258  Comparison of the length of horse metacarpal bones from Marvel with the items from Kernav , Vilnius (the 14ththe first half of the 15th century), Vilnius (the second half of the 15ththe 17th century), Pakalnikiai, Obeliai, Taurapilis, Plinkaigalis, Degsn  Labotik s showed that according to Wil-cocksons criterion the differences of the length of the horse metacarpal bones from these localities were statistically significant (z=-3.779, p<0.001). Comparison of the length of metatarsal bones from Marvel  with the items from Kernav , Vilnius (the 14ththe first half of the 15th century), Vilnius (the second half of the 15ththe 17th century), Pakalnikiai, Obeliai, Taurapilis, Plinkaigalis, Degsn   Labotik s showed that according to Vilkoksons criterion the differences of the length of the horse feet bones from these localities were statistically significant (z = -3.779, p<0.001). According to the relative width of metacarpal bones (Table 3), horses of type III were dominant in Kernav  though remains but all five types of horses were found. The data about Vilnius in the 14ththe first half of the 15th century were rare however they contain evidence about types IIII of horses. The material about Vilnius in the second half of the 15ththe 17th century contained remains of horse types III and IV. Only three metacarpal bones were uncovered in Obeliai. According to their size they belonged to horse type III. The horses buried in Taurapilis were of type IV, in Plinkai-galis III and IV, in Pakalnikiai dominated type III, and in Degsn   Labotik s types II and V. According to the relative width of metatarsal (Table 4), type II and III of horses were dominant in Marvel and Kernav . The same types were found in the material from Vilnius (the second half of the 15ththe 17th century). Types II and III were dominant in Obeliai and 9  
Pakalnikiai, IV in Taurapilis, III in Plinkaigalis, and II in Degsn   Labotik s. The values of the length (GL) of metacarpal and metatarsal bones (Fig. 2) and the thinnest area of diaphysis (SD) arranged on different axes (Fig. 1) yielded the dispersion pattern of individual bone measurements. The data were widely dispersed. The data of individuals (points in the set) differed considerably from the data of main limb bone massifs. This implies that burials contained remains of different types of horses. The correlation coefficient of metacarpal bones length and width of diaphyses was high (r=0.68, p<0.01). The statistically correlation of length of metacarpal bones and width of diaphyses were significant (r=0.56, p<0.01).  Table 3. Relative width of metacarpal bones diaphysis (SD/GL)  Vilnius, Vilnius, Degsn LocalitynKaevr - vMealr - 11st4o thfh a lf sh1ea5cltfoh nofd  ObeliaiTpaiulirs-  kaPilgina-li snPiakkal--Labo- a iai tik s 15th c. 17th c. Width o diaphy-sis (SD/GL) accord-ing to Brauner Type I (up to 1 7,7 1 1 1 25 13.5%) Type II (13.5-1 7,7 9 12 2 50 4 26,7 14.5%) Type III (14.5-8 61,5 25 33,8 1 25 5 62,5 3 100 1 33,3 6 40 2 66,7 15.5%) Type IV (15.5-2 15,4 29 39,2 3 37,5 2 100 2 66,7 4 26,7 16.5%) Type V (16.5-1 7,7 10 13,5 1 6,6 1 33,3 17.5%) Total 12 100 74 100 4 100 8 100 2 100 2 100 3 100 15 100 3 100  10
Table 4. Relative width of metatarsal bones diaphysis (SD/GL)  Vilnius, Vilnius, 14th -Locality nKaevr - vMealr 1sto hf a lf 21d5 othfh alf Obeliai aTpaiulri-s  kaPilginal-isnPiakkaila-i-D tiLeagksb nos - 15th c. 17th c. Width of dia-physis (SD/GL) accord-ing to Brauner Type I (up to 3 25,0 4 8.2 1 50 2 28,6 1 50 11%) II 5 41,7 25 1 50 2 100 (T1y2p%e ) 20 40,8 2 28,6 1 Type III (12-4 33,3 22 44,9 1 50 3 42,8 3 75 1 33,3 1 50 1 50 13%) Type IV (13- 3 6,1 2 66,7 14%) Total 12 100 49 100 2 100 7 100 4 100 3 100 2 100 2 100 2 100  Metacarpal and metatarsal bones were the most abundant and the best preserved items of archaeological material. Therefore, determining the height of horses was not a difficult task. The Lithuanian archaeological ma-terial from the 5th17th c. contained remains of horses whose wither height ranged from less than 112 cm to 160 cm (Table 5).  Teeth measuring  Radiographic and topographic images of 26 present horses were made order to find out which of the methods was more helpful in teeth measuring. Assessment of obtained measuring data by Kolmogorov  Smirnov test re-vealed that normal distribution was obtained only for measuring tooth length p>0.5 and tooth height p>0.5 in computer tomograms. Statistical signifi-cance of the differences between measuring values from tomograms and roentgenograms was determined by Vilkoksons criterion. The obtained statistical significance value of the differences between tooth length values from roentgenograms and tomograms was p<0.001. 11
Metacarpus
40 35 30 25 20 170 180 190 200 210 220 230 240 250 260 Bone lenght, mm Vilnius 14 th -1 st half of 15 th c. Vilnius 2 d half of the 15 th - 17 th c. Kernav Marvel Pakalnikiai   Fig. 1. Length of horse metacarpal bones (GL) versus width of hand diaphyses (SD)   Metatarsus 40 35 30 25 20 15 190 200 210 220 230 240 250 260 270 280 290 300 Bone lenght, mm Vilnius 14 th - 1 st half of 15 th c. Vilnius 2 d half of the 15 th - 17 th c. Kernav  Marvel Pakalnikiai   Fig. 2. Length of horse metatarsal bones (GL)  versus width of feet diaphyses (SD) 12
Statistical significance of the differences of tooth height data was p<0.001. The difference between the average tooth length values in roent-genograms and tomograms was 0.31 mm and as well as the average values of tooth height was 0.48 mm. The obtained results showed that teeth measuring results obtained from roentgenograms were more accurate. The tomographic method was also applicable but the average error of tooth width obtained by this method ranged from 2.8 to 8.7% and the average error of tooth length ranged from 7.7 to 11.5%.  Table 5. Distribution of data in horse wither height, %  ilnius taHoc eciVogirthtdt,i ng nKaevr- Pakal-Obeliai raTpaiuli-s Plgianlkisa iDegsn V14th2Vidl nhiaulsf  - r-Laboti 1st half o cm  nikiaik s 15tohf 1175tthfh  c.vMeal  c. <112 4 25 20 112120, very 16 20 22,2 6,2 36,1 small s1m20all 128, 32 20 50 80  60 22,2 12,5 52,8 128136, smaller 24 13,3 25 20 66,6 20 33,3 31 than av- ,3 11,1  erage a1v3e6ra1g4e4, 24 46,7   3,3  22,2 25   144152, larger       12,5  than av-erage 152160, 12 5 large , n 25 15 4 5 3 5 9 16 72  Determining biological age according to the height of molars  The determining of horses age was based on tooth height values obtained from radiographic and computer tomography images using calliper and re-13
count using A. Levine (1982) method (Table 6). All molars in the lower jaw of 26 present horses (whose age was known) were measured. The smallest difference of biological horse age determined according to the height of second molars  (P2) from the chronological age determined by measuring the teeth with a calliper was 2 years and the greatest difference 2.87 years. The correlation coefficient between the chronological and bio-logical age determined by measuring with a calliper was also high (r=0.809, p<0.01). The chronological age and biological age determined based on to-mograms were in no statistically significant correlation. The greatest difference of biological age average determined according to the third molars (P3) on tomograms was 2.4 years. The correlation coeffi-cient between the chronological age and the age determined using a trammel was high (r=0,854, p<0.01). The chronological age and age determined by teeth measuring on roentgenograms statistically significantly correlated (r=0.785, p<0.01). The correlation coefficient between the chronological age and age determined by tooth height on tomograms showed strong correla-tion of data r=0.723, p<0.05. The difference between the age determined according to the tooth height of fourth molars using a calliper and the chronological age was 2.05 years. The greatest difference was determined when the age was evaluated by measuring teeth on tomograms 2.20. The correlation coefficient between the chronological age and age determined by measuring teeth with a tram-mel was high (r=0.835 p<0.01). The correlation coefficient between the chronological and biological age determined by a trammel was also high (r=0.835, p<0.01). The difference between the average values of chronological age of first molars and biological age determined by measuring on tomograms was 2.87 years. The correlation coefficient between the chronological age and the age determined using a calliper was high (r=0.827, p<0.01). Statistically signifi-cant correlation was determined between the chronological age and age de-termined by measuring teeth on roentgenograms (r=0.827, p<0.01). The correlation coefficient between the chronological age and age determined by teeth measuring on tomograms was also high (r=0.777, p< 0.01). The absolute difference of chronological age of the second molars from the biological age determined by roentgenograms was 1.9 years and from the biological age determined by tomograms 2.4 years. The chronological and biological age determined by teeth measuring in all cases was in strong correlation. 
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Table 6. Results of determining the biological age of animals and dif-ferences from the chronological age  StatiChro-Diffference Hr fDroioflmfe rceihcnracole-  Ht fDrioffme rence nolo- rom chro-stical index gical Hs clhorgoicnaol- naogge nological age age age Average(P2) 8,5 6,88 2,00 6,85 2,08 7,20 2,87 N 26 26 26 26 26 15 15 Min 2,5 3 0 2 0 4 0 Max 17 13 10 14 9 11 9 Standard deviation 4,40 2,93 2,54 3,02 2,46 2,43 2,81 Average (P3) 9,76 8,86 1,29 9,14 1,57 9,40 2,40 N 21 21 21 21 21 10 10 Min 5 5 0 6 0 7 0 Max 17 15 8 15 8 16 8 Standard deviation 3,94 3,14 2,28 3,21 2,38 2,72 2,55 Average (P4) 9,76 8,38 2,05 8,24 1,95 9,00 2,20 N 21 21 21 21 21 10 10 Min 5 4 0 4 0 5 0 Max 17 16 9 15 9 15 8 Standard deviation 3,94 3,38 2,13 3,56 2,20 2,83 2,57 Average (M1) 8,5 6,88 2,00 6,85 2,08 7,20 2,87 N 26 26 26 26 26 15 15 Min 2,5 3 0 2 0 4 0 Max 17 13 10 14 9 11 9 Standard deviation 4,40 2,93 2,54 3,02 2,46 2,43 2,81 Average (M2) 9,76 8,43 1,76 8,29 1,90 8,70 2,40 N 21 21 21 21 21 10 10 Min 5 4 0 4 0 5 0 Max 17 14 9 14 9 13 8 Standard deviation 3,94 3,30 2,02 3,33 2,07 2,71 2,41 Average (M3) 9,76 9,00 2,00 9,14 2,33 9,00 3,00 N 21 21 21 21 21 10 10 Min 5 3 0 3 0 6 0 Max 17 20 9 20 9 13 9 Standard deviation 3,94 4,02 2,63 4,34 2,69 2,62 3,23  Hs  biological age according to tooth height measured with the calliper; Hr  biological age according to tooth height measured on radiographic images; Ht  biological age according to tooth height measured on computer tomography images.
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The greatest difference of the biological age of the third molars from the chronological age was obtained when teeth were measured on topographic images (3 years) and the smallest when teeth were measured using a calliper (2 years). The correlation coefficient between the chronological age and biological age determined by a calliper was high (r=0.719 p<0.01). There was no correlation between the chronological and biological age determined by measuring teeth on computer tomography images. The presented data showed that the smallest differences from the chrono-logical age were obtained when teeth were measured using a calliper. The greatest differences were obtained when teeth were measured on computer tomography images.  Determining the age of excavated horse according to the height of molars The age of all excavated horses was determined according to the wear of incisors (WI) and according to the shift from milk-teeth to permanent teeth (MPTS). The age of horses was determined based on tooth height values obtained from radiographic and computer tomography images measuring data obtained using a calliper and counted by A. Levine (1982) method (Ta-ble 7). All molars in the lower jaw of 16 exacvated horses were measured. The difference of age of horses determined by measuring the second mo-lars (P2) with a trammel and on roentgenograms from the age determined according to WI and MPTS was 2.13 years. The difference between the age determined by measuring teeth on a tomogram and the chronological age was 2.33 years. The correlation coefficient between the age determined ac-cording to WI and MPTS and the age determined by teeth measuring showed no correlation between these values p>0.05. However the data of individual measuring were in statistically significant correlation r=0.921, p<0.01. The smallest difference of 3.13 years was obtained between the age of the third molars determined according to WI and MPTS and the age deter-mined with the aid of a calliper. The greatest difference was obtained when teeth were measured on tomograms  3.56 years. The age determined ac-cording to WI and MPTS is in no correlation with the age determined by measuring the teeth height p>0.05. The greatest difference of 3.25 years was obtained between the age of the fourth molars (P4) determined according to WI and MPTS and the age determined by measuring tooth height on radiographic images. The age de-termined according to WI and MPTS is in no correlation with the biological age determined by measuring the teeth height p>0.05.  
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Table 7.  Results of determining the biological age of excavated horses and differences from the chronological age  - Difference Difference Age de Difference from the age from the Statistical atcetcroom riWdniIendg  Hamfgrieon emdde  ttahecre-- aged idnetge tr-Hr determined index s ding to according to Ht mined ac-and coWrI and WI and coWrI and o MPTS MPTS MPTS MPTS Average(P2) 8,25 9,60 2,13 9,87 2,13 9,73 2,33  16 15 15 15 15 15 15 Min 5 6 0 6 0 6 1 Max 16 15 8 15 8 15 8 STDV 3,70 3,07 1,88 3,09 1,96 2,96 1,88 Average (P3) 8,25 9,88 3,13 10 3,38 9,94 3,56  16 16 16 16 16 16 16 Min 5 6 0 6 1 6 0 Max 16 20 15 20 20 20 15 STDV 3,69 4,19 3,99 3,93 3,86 3,94 3,86 Average (P4) 8,25 9,50 3,13 9,75 3,25 9,50 3,13  16 16 16 16 16 16 16 Min 5 6 0 6 0 6 0 Max 16 20 15 20 15 20 15 STDV 3,69 3,67 3,84 3,69 3,80 3,62 3,76 Average(M1) 8,25 9,13 2,38 9,00 2,50 9,06 2,69  16 16 16 16 16 16 16 Min 5 6 0 6 0 5 0 Max 16 18 13 17 12 18 13 STDV 3,70 3,32 3,12 2,97 2,78 3,19 3,16 Average(M2) 8,25 9,69 2,69 9,19 2,94 9,38 2,94  16 16 16 16 16 16 16 Min 5 6 0 5 0 6 0 Max 16 15 9 15 9 15 9 STDV 3,69 2,77 1,96 2,76 2,32 2,55 2,32 Average(M3) 8,25 11,4 4,13 11,27 4,13 11,8 4,40  16 15 15 15 15 15 15 Min 5 7 1 7 1 8 2 Max 16 20 14 20 14 20 14 STDV 3,69 4,05 3,14 4,20 3,18 3,84 3,02 Hs  biological age according to tooth height measured with the aid of calliper; Hr  biological age according to tooth height measured on radiographic images; Ht  biological age according to tooth height measured on computer tomography images. 17
The difference between the ages of the first molars (M1) determined accord-ing to WI and MPTS and the age determined by measuring teeth height with a calliper was 2.38 years. There were no correlation between the age of the first molars (M1) determined according to WI and MPTS and the biological age determined by teeth measuring p>0.05. The greatest difference of 2.94 years was obtained between the age of the second molars (M2) determined according to teeth height measuring on radiographic and computer tomography images. The average biological age determined by measuring with a calliper is 9.69 years. Its difference from the age determined by WI and MPTS was 2.69 years. Statistically signifi-cant correlation was established only between the age determined according to WI and MPTS and biological age determined by measuring with a tram-mel r=0.635, p<0.01. There were no correlation between the age determined according to WI and MPTS and the biological age determined by measuring the teeth height on tomograms, p>0.05. The difference between the average values of the age determined accord-ing to WI and MPTS and biological age determined by measuring the third molars (M3) with a calliper and on radiographic images was 4.13 years. There were no correlation between the age determined according to WI and MPTS and the biological age determined by teeth height measuring p>0.05. The greatest determined difference was between the age according to WI and MPTS and the biological age determined on the basis of tomograms.  Counting of annual cementum rings in the teeth microstructure of present horses The main research results showed in Table 8. The annual cementum rings were counted on the teeth of 26 present horses based on 30 histological samples. The results of three counting methods of cementum rings corre-lated very high (r>0.968, p<0.01). This meaned that counting results of an-nual cementum rings were very objective and differences of counting pro-duced no major effect on the data. Therefore the correlation coefficient be-tween the age determined by annual rings and chronological age was high r=0.94, p<0.01. The absolute difference of the data set from the chronologi-cal age was 0.76 years on the average. The accuracy of age determined for older animals decreased; the difference of the whole data set was 0.76 years (8.78%). For young (up to 4 years of age) individuals this value was smaller  0.54 years (6.32%), for individuals aged up to 10 years 0.73 years (8.54%) and for individuals aged more than ten years 0.88 (10.3%).  
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Table 8. Results of determining biological age of present horses and 18 difference from the chronological age  16 Statistical indexChrono-vAavlerea goef  Determined Absoel uftreo dmi ftfheer -14  logical cemeuntum bae age rings iological gchernocnological age Average (all) 8,54 8,13 8,63 0,76 12 N 26 26 26 26 Min 2,5 1,3 1,8 0 10 Max 17 15,7 16,2 2,8 Standard deviation 4,40 4,44 4,43 0,84 8 Average (up to 4 years) 3,2 2,16 2,66 0,54 6 N 5 5 5 5 Min 2,5 1,3 1,8 0 4 Max 4 3,3 3,8 1,7 Standard deviation 0,57 0,77 0,77 0,73 2 Average (410 years) 6,55 6,66 7,16 0,73 0 2 4 6 8 10 12 14 16 N 11 11 11 11  Min 5 5,1 5,6 0  Max 9 11 11,5 2,8 Fig. 3. Mutual dependence of annual cementum rings on the teeth of Standard deviation 1,04 1,57 1,57 1,01 present horses (axis X) and chronological age (axis Y) . Broken line marks Average individual cases and solid line stands for cubic regression. (more than 10 years) 13,4 12,74 13,22 0,88  N 10 10 10 10 Min 10 8,5 9 0 and T d a i b f l f e e  r 9 e . n cRe efsruoltms  tohf e dcehtreornmoilnoignigc abl iaogleo g ical age of excavated horses Max 17 15,7 16,2 2,2  Standard deviation 2,37 2,37 2,40 0,76  Absolute dif- Arithmetic  Age Dependence between the number of cementum rings and chronological accord- Average Deter- ference from difference age used a non-linear character: the highest correlation was obtained using Statistical index iWnIg  atnod  cneummenetru omf lmoiniecda l baigoe-ctotehrrdem iainngge et dod  aec-W-I fardcoectmoe rrtdhmiein ngae gtdoe    cubic (Y = b 0 +b 1 X+b 2 X 2 +b 3 X 3 ) (Fig. 3) regression model (r=0.943). MPTS rings g d MPTS WI and MPTS   an Counting of annual cementum rings in the tooth microstructure of Average(all) 10,81 10,58 11,08 0,68 -0,26 excavated horses  N 8 8 8 8 8 The age of excavated horses was determined according to the shift of Min 5 4,3 4,8 0 -2,2 milk teethpermanent teeth (MPTS) and wear of incisors (WI). Cementum Max 16 15,5 16 2,2 1,2 rings were counted in 8 histological samples (40%). The obtained results Standard deviation 4,72 4,86 4,86 0,91 1,24 (Table 9) correlated highly. The correlation coefficient is r>0.922, p<0.01. Different r i n models showed  that the determined interdependence The ages determined by counting cementum rings and by WI and MPTS used a n-lineegarr ecshsaroacter: the highest correlation was obtained using cubic also correlated highly r>0.946, p<0.01. The average difference between the on ages determined according cementum rings for the whole data set and ac- regression models (Y = b 0 +b 1 X+b 2 X 2 ) (Fig. 5.) (r =  0.942) and (Y = cording WI and MPTS was 0.68 years. b 0 +b 1 X+b 2 X 2 +b 3 X 3 ) (Fig. 4). 19 20
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