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Within the Bosnian Mountain Horse Registry (BMH), some specific coat colours are permitted. Accurate determination of coat colours can be challenging as there are variations in coat colour shades and several dun dilution variants occur. The previously found single nucleotide polymorphisms (SNPs) within two coat colour loci, T-box 3 (TBX3) and Agouti Signalling Protein (ASIP), were genotyped in 313 BMH individuals. The obtained genotypes were then compared to the predicted phenotypes by using the observed coat colour types from the International Association of Bosnian Mountain Horse Breeders (IABMHB) database. It was found that the dark bay and black coat colours were the most representative coat colours both phenotypically and genotypically. The frequency of the dominant Dun (D) dilution allele is (0.09), and this frequency is higher than the previously predicted frequency recorded in the available BMH registers, as also demonstrated. Among the identified alleles, there was a discrepancy or inconsistency between the predicted coat colour based on genotypes and the observed coat colour in 73 horses (23%). The most frequent error concerned the misclassification of horses with genotypes aa and Aa at the ASIP gene, non-dun1/non-dun1 (nd1/nd1) and non-dun2/non-dun1 (nd1/nd2) at the TBX3 gene, which can be associated with the occurrence of slight dilution phenotypes in these individuals. In contrast to the Konik and Hucul breeds, no homozygosity of the D allele was found in the BMH. The D allele can be easily overlooked or not recognised in different phenotypic groups, such as dark bay and black horses. Therefore, the hypothesis that Dun dilution effects itself is not as strongly epistatic in the BMH as described in other horse breeds. This could be the result of an additional genetic modifier suppressing the phenotypic effect of the D allele. It also suggests that there has been a persistent selection pressure in favour of dark base colours, which may have contributed to the observed differences in BMH. Finally, a significant proportion (35%) of BMH individuals with genetically black coat colour were officially classified as dark bay. We hypothesise that the difficulty in visually distinguishing these two phenotypes is due to an independent locus upstream of the ASIP gene. This locus was recently identified as a factor that darkens the typical pigmentation in dark bay horses. The results of the study confirm the importance of molecular testing in accurately determining the coat colour of horses. This would help to avoid errors in coat colour descriptions in official breeding records and provide valuable information for selective breeding programmes aimed at producing specific and desired coat colours. Variabilnost genov ASIP in DUN, odgovornih za barvo dlake pri bosanskem planinskem konju Določanje barv dlake pri bosanskem planinskem konju (BPK) je lahko izziv, ker obstajajo razlike v barvnih odtenkih dlake in več oblik redčenja barve dlake (plavci). Pri 313 konjih pasme BPK smo genotipizirali polimorfizme posameznih nukleotidov (SNP) znotraj dveh lokusov za barvo dlake T-box 3 (TBX3) in polimorfizma 11-bp indel znotraj Agouti signalnega proteina (ASIP). Posamezne genotipe smo nato primerjali z vpisanimi fenotipi barv dlake iz baze podatkov MZRBPK (Mednarodno združenje rejcev bosanskega planinskega konja). Ugotovljeno je bilo, da sta bili temna rjava in črna najbolj reprezentativni barvi dlake pri BPK. Pogostnost prevladujočega alela za redčenje Dun (D) v študiji je višja (0,09) od predhodno zabeležene v razpoložljivem registru BPK. Med ugotovljenimi aleli je prišlo do neskladja ali nedoslednosti med predvideno barvo dlake na podlagi genotipov in opazovano barvo dlake pri 73 konjih (23%). Najpogostejša napaka se je nanašala na napačno razvrstitev konjev z genotipoma aa in Aa v genu ASIP, non-dun1/non-dun1 (nd1/nd1) in non-dun1/non-dun2 (nd1/nd2) pri genu TBX3, kar je lahko povezano s pojavom redčenja barve dlake pri teh fenotipih.V nasprotju s pasmami Konik in Hucul pri BPK ni bila ugotovljena homozigotnost alela D. Alel D je mogoče zlahka spregledati ali ga ne prepoznati v različnih fenotipskih skupinah, kot so temni rjavci in vranci. Zato hipoteza, da učinki redčenja Dun (plavci) pri BPK sami po sebi niso tako močno epistatični, kot je to opisano pri drugih pasmah konj. Rezultati študije potrjujejo pomen molekularnega testiranja pri natančnem določanju barve dlake konj. To bi pripomoglo k preprečevanju napak pri opisih barve dlak v uradnih rejskih evidencah in pomembna informacija pri selekciji na posebne in želene barve dlake pri konjih. Ključne besede: barva dlake konj; dun redčenje barve dlake; polimorfizem DNK; frekvenca alelov; frekvenca genotipa; bosanski planinski konj

The aim of this study was to investigate the influences of sex and age on haematological values in the Posavje Horse breed. A total of 163 healthy Posavje horses (30 foals, 94 mares and 39 stallions) were used in this study; their complete blood counts and a leucogram were obtained with a haematological analyser. The horses were classified into five groups: foals (1 to 6 months, n = 30), 3 to 6 years (n = 8 stallions/21 mares), 7 to 9 years (n = 9 stallions/22 mares), 10 to 13 years (n = 8 stallions/20 mares), 14 to 15 years (n = 6 stallions/10 mares) and 16 and over (n = 8 stallions/21 mares). The results obtained show an influence of sex on haematological parameters, with red blood cell count (RBC), haematocrit (HCT) and haemoglobin concentration (HGB) being higher in stallions (P < 0.001) and white blood cell count (WBC) being higher in mares. Differences between the age groups of the Posavje horses examined indicate a decrease in RBC and HGB with a compensatory increase in mean corpuscular volume and mean corpuscular haemoglobin, a decrease in WBC and platelet counts (PLT) and proportion of lymphocytes, and an increase of neutrophil to lymphocyte ratio (N/L) with age (P < 0.001). Although the Posavje horse is classified as a draft horse breed, its haematological parameters show characteristics common to warm-blooded breeds, with the exception of the N/L ratio. One of the most important findings of this study is a higher neutrophil count in reproductively active breeding stallions. Higher levels of RBC, HGB, HCT and neutrophil count in the Posavje stallions suggest an effect of androgens (testosterone), which may be an effective mechanism to prevent infections, that can affect the survival of the stallions and thus the evolution of the species. KARAKTERIZACIJA HEMATOLOŠKEGA PROFILA PRI POSAVSKEM KONJU Izvleček: Cilj raziskave je bil proučiti vpliv spola in starosti na hematološke parametre pri pasmi posavski konj. V raziskavo je bilo vključenih 163 konj posavske pasme (30 žrebet, 94 kobil in 39 žrebcev), pri katerih smo v vzorcih krvi določali hematološke parametre s hematološkim analizatorjem. Diferencialna bela krvna slika in razmerje med nevtrofilci in limfociti (N/L) je bilo določeno na krvnih razmazih. Konje smo razdelili v pet starostnih skupin: žrebeta (od 1 do 6 mesecev, n = 30), 3 do 6 let (n = 8 žrebcev/21 kobil), 7 do 9 let (n = 9 žrebcev/22 kobil), 10 do 13 let (n = 9 žrebcev/20 kobil), 14 do 15 let (n = 6 žrebcev/10 kobil) ter 16 in več let (n = 8 žrebcev/21 kobil). Rezultati naše raziskave kažejo vpliv spola na preiskovane hematološke parametre; pri žrebcih so število rdečih krvnih celic (RBC), hematokrit (HCT) in koncentracija hemoglobina (HGB) značilno višji (P < 0,001), pri kobilah pa je višje število belih krvnih celic (WBC). Med starostnimi skupinami posavskih konj smo ugotovili zmanjšanje RBC in HGB in posledično kompenzacijo s povečanjem povprečnega volumna in hemoglobina eritrocitov, zmanjšanjem števila levkocitov, trombocitov (PLT) in limfocitov ter povečanjem razmerja med nevtrofilci in limfociti (N/L) s starostjo (P < 0,001). Posavski konj po zunanjosti spada med hladnokrvne konje, v raziskavi ugotovljeni hematološki profil pa kaže značilnosti, ki so skupne toplokrvnim pasmam konj, z izjemo razmerja N/L. Ena od pomembnejših ugotovitev te študije je večje število nevtrofilcev pri aktivnih plemenskih žrebcih. Višje vrednosti RBC, HGB, HCT in števila nevtrofilcev pri posavskih žrebcih kažejo učinek androgenov (testosterona), kar bi lahko bil učinkovit mehanizem za preprečevanje okužb, ki lahko vplivajo na preživetje žrebcev in s tem na evolucijo vrste. Ključne besede: avtohtone pasme; posavski konj; hematologija; starost; spol

Different approaches and classification systems have been established to describe equine coat colour, which varies between breeds and countries. In the present study, we first characterised the coat colour variability in the Slovenian Posavje Horse applying colour spectrophotometry following the CIE L*a*b system. As derived from the classification system of Sponenberg (light bay, bay, mahogany bay, brown and seal brown), the phenotype categories could be confirmed by spectrophotometric data. Furthermore, L*a*b values revealed comparable high phenotypic variability of bay coat colour in the Posavje breed, and the darker shades of bay coat colour were associated with the ASIP and MC1R genotype combination A/a E/E. CIE L*a*b colour spectrophotometry represents an effective tool to characterise and quantify coat colour in horses, especially in chestnut horses, for which the underlying genetic background of coat colour variation remains unknown. Key words: Posavje Horse; MC1R; ASIP; coat colour; spectrophotometry; CIE L*a*b KARAKTERIZACIJA BARVE DLAKE PRI POSAVSKEM KONJU Izvleček: Za opis barv konj se uporabljajo različni pristopi in klasifikacijski sistemi, ki se razlikujejo med posameznimi pasmami in državami. V raziskavi smo najprej opredelili različne barve dlake pri posavskem konju z metodo barvne spektrofotometrije po sistemu CIE L*a*b*. Fenotipsko razdelitev barv dlake po Sponenbergu (light bay/svetli rjavec, bay/rjavec, mahogany bay/kostanjev, brown/temni rjavec and seal brown/črnkast rjavec) smo potrdili s spektrofotometričnimi podatki. Vrednosti L*a*b so pri posavskem konju pokazale primerljivo visoko fenotipsko variabilnost rjave barve, pri tem so bili temnejši odtenki povezani z ASIP in MC1R kombinacijo genotipa A/a E/E. Barvna spektrofotometrija po sistemu CIE L*a*b predstavlja učinkovito orodje za kvalitativno in kvantitativno opredelitev/določanje barv pri konjih, zlasti pri lisjakih, pri katerih še vedno ni znana genska osnova variabilnosti v barvi dlake. Ključne besede: posavski konj; MC1R; ASIP; barva dlake; spektrofotometrija; CIE L*a*b

Slovenian Cold-Blooded horse is an autochthonous horse breed in Slovenia, traditionally reared in the North-Eastern and Northern parts of Slovenia. Today the breed is widespread all over the country. Breeding program for the Slovenian Cold-Blooded horse was accepted 2005 when the Association of Slovenian Cold-Blooded Horse Breeders was established, too. The aim of the study was to analyse conformation and gaits traits of the Slovenian Cold-Blooded horse. Likewise, we tried to evaluate fixed effect, affecting the included traits. Data were collected during the classifications of Slovenian Cold-Blooded horse performed from 1996 to 2011. In this study, 1920 horses were included, 52 of which were stallions and 1868 mares. The scoring system included 8 measured and 10 scored traits. Data were analysed by GLM procedure of statistical package SAS/STAT considering sex, age at scoring and birth year as fixed effects. Stallions of Slovenian Cold-Blooded horses were on the average 152.4±0.56 cm high at withers (stick), while mares were 151.22±0.11 cm. Body length (stallions 163.95±1.48 cm; mares 164.28±0.17 cm) was on the average larger than the height at wither thus indicating the rectangular body frame.

In the nineteenth and early twentieth century the Medžimurje horse played an important role as a draft and working horse in the river Mura region. Nowadays, this area is split between Hungary, Austria, Slovenia and Croatia. In the frame of the project "Management of traditional transboundary breeds on example of a nearly forgotten breed the Murinsulaner", we collected data for cold blooded horses similar to the Medžimurje horse. Horses of this type do not exist in Austria anymore, in Croatia they were registered as an authothonous breed »Medjimurje horse«, and in Slovenia and Hungary this horse has a special section in the stud book for cold blooded horses as a Medžimurje type of horse. The Slovenian part of the population of the Medžimurje horse is quite heterogeneous regarding body measures. With respect to the scoring system used in the above mentioned project, the majority of scored animals were classified as »desired« or »satisfactory«. In order to further develop or to preserve this type of horse or to form a consolidated breed we would have to make a breeding program which would prevent mating among related animals and to cooperate with neighbouring countries in the field of breeding and selection. The only hope to preserve this type of horse on the long term in the region is by using it as a tourist attraction for carriage riding and/or as a draft horse in agriculture in marginal regions.