The importance of local breeds as genetic reservoirs of valuable genetic variation is well established. Pig breeding in Central and South-Eastern Europe has a long tradition that led to the formation ...of several local pig breeds. In the present study, genetic diversity parameters were analysed in six autochthonous pig breeds from Slovenia, Croatia and Serbia (Banija spotted, Black Slavonian, Turopolje pig, Swallow-bellied Mangalitsa, Moravka and Krskopolje pig). Animals from each of these breeds were genotyped using microsatellites and single nucleotide polymorphisms (SNPs). The results obtained with these two marker systems and those based on pedigree data were compared. In addition, we estimated inbreeding levels based on the distribution of runs of homozygosity (ROH) and identified genomic regions under selection pressure using ROH islands and the integrated haplotype score (iHS).
The lowest heterozygosity values calculated from microsatellite and SNP data were observed in the Turopolje pig. The observed heterozygosity was higher than the expected heterozygosity in the Black Slavonian, Moravka and Turopolje pig. Both types of markers allowed us to distinguish clusters of individuals belonging to each breed. The analysis of admixture between breeds revealed potential gene flow between the Mangalitsa and Moravka, and between the Mangalitsa and Black Slavonian, but no introgression events were detected in the Banija spotted and Turopolje pig. The distribution of ROH across the genome was not uniform. Analysis of the ROH islands identified genomic regions with an extremely high frequency of shared ROH within the Swallow-bellied Mangalitsa, which harboured genes associated with cholesterol biosynthesis, fatty acid metabolism and daily weight gain. The iHS approach to detect signatures of selection revealed candidate regions containing genes with potential roles in reproduction traits and disease resistance.
Based on the estimation of population parameters obtained from three data sets, we showed the existence of relationships among the six pig breeds analysed here. Analysis of the distribution of ROH allowed us to estimate the level of inbreeding and the extent of homozygous regions in these breeds. The iHS analysis revealed genomic regions potentially associated with phenotypic traits and allowed the detection of genomic regions under selection pressure.
Meat from Black Slavonian pig (Crna slavonska, CS), an autochthonous Croatian breed, is used for the production of high-quality products, which makes it an interesting target for meat fraud. The aim ...of the study was to develop a set of microsatellite markers suitable for precise genetic traceability of the meat originating from this breed. Initially, 23 microsatellite markers were selected and grouped into three multiplex reactions for the genetic characterization of the seven breeds included in the study; subsequently, a set of eight microsatellite markers for traceability of CS pig was created based on the match probability value. The results indicated that when eight highly polymorphic loci were combined, the chance of finding identical genotype in two random individuals was about three in ten million (10−7). This provided the basis for establishing a reliable genetic traceability system for meat originating from CS pigs.
•A microsatellite set was selected for animal identification of Black Slavonian (Crna slavonska) pig breed.•The set was evaluated and further optimised using Match Probability (MP) value.•The set was validated for meat traceability of Black Slavonian pig breed.
The Turopolje pig (TP) is a local Croatian pig breed that almost became extinct in the second half of the 20th century. Today, the TP is still endangered, and a new conservation strategy based on ...products with higher added value is needed to preserve the breed. There is little information on the quality of TP meat products such as smoked and dry-cured ham, including the impact of natural feeds or processing innovations such as smoke reduction. This study, therefore, investigated the effects of the animal's diet (either conventionally fed or acorn-supplemented) and the processing method (standard or lightly smoked) on the quality traits of dry-cured TP ham. Twenty hams, evenly distributed among the treatments, were processed for 15 months and then analysed for physicochemical and textural traits, volatiles and sensory profile. The hams from acorn-supplemented pigs lost less weight during processing (
≤ 0.05). Otherwise, the diet had no significant effect on most examined ham traits. The exceptions were protein content and the texture parameter hardness, which decreased (
≤ 0.05), and the degree of proteolysis and colour parameters, which increased (
≤ 0.05) as a result of acorn supplementation. However, these effects were generally small and varied between the inner (
) and outer (
) muscles. Furthermore, acorn supplementation was associated with less typical ham odour and lower sensory scores for sweetness and colour uniformity (
≤ 0.05). The smoke reduction had no effect on the physicochemical and colour properties but resulted in a significant reduction (
≤ 0.05) in the volatile phenolic compounds and an improved texture to the hams. This was reflected both in reduced (
≤ 0.05) hardness, identified in the instrumental analysis, and in an increased (
≤ 0.05) softness, solubility and moistness, identified in the sensory evaluation. To summarize, the quality of the TP ham under the conditions studied was only slightly affected by acorn supplementation, whereas reduced smoking had a more significant effect, which was mainly reflected in an improved texture.
The aim of this work was to evaluate the influence of cheese milk standardization on chemical composition, fat and protein recovery, yield and sensory properties of Croatian soft Protected ...Geographical Indication (PGI) Lički škripavac cheese. Standardization of milk to the casein/fat ratio of 0.7 was carried out by adding skimmed milk powder (SMP) to cheese milk and by skimming part of the milk fat. Results showed that losses of fat by whey were significantly (
< 0.05) lower after Lički škripavac cheese produced from standardized milk by skimming part of the milk fat. Standardization of cheese milk by addition of SMP caused higher losses of protein (
< 0.05) and total solids (
< 0.0001) by whey. Both methods of cheese milk standardization caused a significant (
< 0.01) decrease in milk fat and fat in dry matter content in cheese. In contrast, standardization of cheese milk caused a significant (
< 0.01) increase in protein content in cheese milk. Moisture in non-fat substance (MNFS) significantly (
< 0.05) decreased. Optimization of the casein/fat ratio did not cause a significant increase in fat recovery, but protein recovery significantly increased (
< 0.01). Addition of SMP to cheese milk significantly (
< 0.01) increased actual and adjusted cheese yield. The addition of SMP led to a noticeably higher (
= 0.10) sensory score of Lički škripavac cheese.
Black Slavonian (DBS) pig and Turopolje pig (DTP) are endangered autochthonous Croatian pig breeds that are still reared in a traditional way under semi-wild conditions in an area populated with ...growing wild boar populations. This raises the possibility of hybridization between domestic pigs and wild boars, a possibility often debated but rarely analyzed genetically. We analyzed potential hybridization between populations of domestic pigs (DBS, N=28; DTP, N=46) and wild boars (WBS) in eastern and western Slavonia (N=20 each) using 14 microsatellite markers and a Bayesian clustering algorithm (STRUCTURE). The STRUCTURE analysis classified all animals into corresponding WBS, DBS and DTP clusters. However, the DTP cluster contained four DTP animals with WBS membership coefficients (q) ranging from 0.25 to 0.35. These results provide genetic evidence of recent gene flow from WBS into DTP. We recommend monitoring the purity of DTP individuals to preserve the genetic integrity of this critically endangered breed. More generally, our findings provide an example of multiple hybridization events in the history of pig domestication.
The microsatellite (MS) genetic diversity studies on CS pig breed showed subclustering of this breed. Structuring results based on MS data of 70 CS pigs were compared to the MC1R coat color gene ...genotyping results. The MS analysis included also commercial pig breeds to elucidate the relationship of different CS breed gene pools towards commercial pig breeds. The Structure results and the results of Unweighted Pair Group Method with Arithmetic Mean (UPGMA) individual clustering based on genetic distances revealed presence of three gene pools in CS breed. Comparison with the genotyping results of the MC1R coat color gene showed that one gene pool consists of MC1R homozygous black animals, while animals from the other two gene pools (CS2 and CS3) were MC1R heterozygous.
This study aimed to determine the relationship between intramuscular fat (IMF) content and fatty acid (FA) composition of pork. The IMF content and the FA composition were determined in the samples ...(n = 30) of muscle tissue (m. longissimus dorsi) taken from the pig carcasses from the intensive pork production system in Republic of Croatia. The fat content was analysed by standard ISO method, while FA composition was determined by liquid gas chromatography using the in situ transesterification method. Results revealed the significant positive correlations and regressions of monounsaturated FA (MUFA) on IMF content, while reverse relationship existed between IMF and the most of the polyunsaturated FA (PUFA). Although there was a significant relation between the IMF content and the majority of individual saturated FA (SFA), the effect of IMF content on the proportion of total SFA in pork was not clear. This can be explained by the existence of significant correlations and regressions but with an opposite directions for major SFA-s, which shares in IMF were simultaneously increasing (e.g. 16:0) and decreasing (e.g. 18:0) with an increase in IMF content.
Banija spotted pig breed was created at the end of 19th century by crossing domestic white sows with drooping ears and/or Turopolje pig sows with black Berkshire. After a period of stagnation in ...breeding, the population is newly recognized only few years ago and is currently in the process of breed valorisation. The aim of this study was to assess genetic diversity and structure of Banija spotted pig (BS) in comparison with geographically close populations, and to assess the differentiation level of Banija spotted pig. Based on the genotypes obtained for 24 ISAG-FAO recommended pig microsatellite markers, basic genetic diversity indicators were determined for 30 samples of BS, 20 samples of Turopolje and Crna Slavonska pig each, and 17 samples of Landrace population. Observed heterozygosity ranged from 0.36 to 0.6 and was high in BS population (0.58). Average allelic richness (range 2 - 5.2) and the rarefacted number of private alleles (range 9 to 27) were the highest in BS. Inbreeding coefficient in BS was estimated 0.06 (bootstrapping confidence interval 0.022 to 0.123). Structure analysis showed that there are five clusters in the four analysed populations, where BS are clearly distinguished from other populations and substructured. Wright's fixation index estimates are in accordance with the clustering results. In conclusion, the population of Banija spotted pig shows great genetic diversity and is genetically different from neighbouring (geographically close) pig populations.
Livanjski cheese belongs to the group of hard cheeses which is traditionally produced in Livno (Bosnia and Herzegovina). Proteolytic changes during the ripening of Livanjski cheese have not been ...investigated extensively. The aim of this paper was to determine its proteolytic changes during the different stages of ripening. Five Livanjski cheeses (from raw cow’s or a mixture of sheep’s and cow’s milk) were observed during the ripening to evaluate its typical proteolytic profile. An electophoretic profile of Livanjski cheese was determined by Urea-polyacrylamide gel electrophoresis (urea-PAGE) and a densitometric evaluation of the urea-PAGE gels was performed using a densitometer. The water-soluble nitrogen fraction in the total nitrogen (WSN %TN) and the 12%-TCA-soluble nitrogen fraction in the total nitrogen (TCA-SN %TN) of the cheese were determined using the Kjeldahl method. Degradation of αs1-casein by chymosin caused a significant decrease (P < 0.05) of relative content of this protein in Livanjski cheese at the sixth week point of ripening. Due to the activity of chymosin on αs1-casein, αs1-I-casein and αs1-II-casein developed, which caused a significant increase (P < 0.05) of Index alpha. The relative ratio of β-casein significantly decreased (P < 0.05) during ripening leading to a significant accumulation (P < 0.05) of degraded product (sum γ1-casein, γ2-casein and γ3-casein). These proteolytic changes caused a significant increase (P < 0.05) of Index betta. Accumulation of medium, small peptides and amino acids caused a significant (P < 0.05) increase of the relative content of WSN %TN and TCA-SN %TN. In general, proteolysis of Livanjski cheese during ripening was moderate probably due to the low moisture content and low water activity, although it was produced from raw milk. Taking into account that the ratio β-casein : αs1-casein at the end of ripening was 1.46, it could be concluded that degradation of αs1-casein could be the indicator of the maturity of Livanjski cheese. Due to that Livanjski cheese could be classified as “α-type of ripening” cheese.
Genetic diversity and breed structure of Banija spotted pig (BS) was analyzed trough 721 pedigree records and polymorphism of 23 microsatellites on 30 BS pigs. Two phylogenetic trees were constructed ...from microsatellite and pedigree information. The inbreeding coefficient obtained by microsatellite markers was 5.6%, while the inbreeding coefficient from pedigree analysis was 3.68%. The rate of inbreeding per generation was 1.74% and effective population size was 28.81. In phylogenetic analyses, relationship coefficients and genetic distances between individuals were calculated using microsatellite and pedigree data. Phylogenetic trees from microsatellite markers and pedigree corresponded well to each other and showed consistency between microsatellite and pedigree information. From the data obtained by microsatellite markers and pedigree, two subpopulations can be observed. The existence of two subpopulations can be explained by two different paths of breed genesis. Thus, future work in the conservation process should include methods such as optimal contribution selection including factorial mating, in order to make genetic progress and control the rate of inbreeding.
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