Runs of homozygosity provide a genome landscape picture of inbreeding and genetic history of European autochthonous and commercial pig breeds.

ROHs are long stretches of DNA homozygous at each polymorphic position. The proportion of genome covered by ROHs and their length are indicators of the level and origin of inbreeding. Frequent common ROHs within the same population define ROH islands and indicate hotspots of selection. In this work, we investigated ROHs in a total of 1131 pigs from 20 European local pig breeds and in three cosmopolitan breeds, genotyped with the GGP Porcine HD Genomic Profiler. plink software was used to identify ROHs. Size classes and genomic inbreeding parameters were evaluated. ROH islands were defined by evaluating different thresholds of homozygous SNP frequency. A functional overview of breed-specific ROH islands was obtained via over-representation analyses of GO biological processes. Mora Romagnola and Turopolje breeds had the largest proportions of genome covered with ROH (~1003 and ~955 Mb respectively), whereas Nero Siciliano and Sarda breeds had the lowest proportions (~207 and 247 Mb respectively). The highest proportion of long ROH (>16 Mb) was in Apulo-Calabrese, Mora Romagnola and Casertana. The largest number of ROH islands was identified in the Italian Landrace (n = 32), Cinta Senese (n = 26) and Lithuanian White Old Type (n = 22) breeds. Several ROH islands were in regions encompassing genes known to affect morphological traits. Comparative ROH structure analysis among breeds indicated the similar genetic structure of local breeds across Europe. This study contributed to understanding of the genetic history of the investigated pig breeds and provided information to manage these pig genetic resources.

Genome-wide detection of copy number variants in European autochthonous and commercial pig breeds by whole-genome sequencing of DNA pools identified breed-characterising copy number states.

In this study, we identified copy number variants (CNVs) in 19 European autochthonous pig breeds and in two commercial breeds (Italian Large White and Italian Duroc) that represent important genetic resources for this species. The genome of 725 pigs was sequenced using a breed-specific DNA pooling approach (30-35 animals per pool) obtaining an average depth per pool of 42×. This approach maximised CNV discovery as well as the related copy number states characterising, on average, the analysed breeds. By mining more than 17.5 billion reads, we identified a total of 9592 CNVs (~683 CNVs per breed) and 3710 CNV regions (CNVRs; 1.15% of the reference pig genome), with an average of 77 CNVRs per breed that were considered as private. A few CNVRs were analysed in more detail, together with other information derived from sequencing data. For example, the CNVR encompassing the KIT gene was associated with coat colour phenotypes in the analysed breeds, confirming the role of the multiple copies in determining breed-specific coat colours. The CNVR covering the MSRB3 gene was associated with ear size in most breeds. The CNVRs affecting the ELOVL6 and ZNF622 genes were private features observed in the Lithuanian Indigenous Wattle and in the Turopolje pig breeds respectively. Overall, the genome variability unravelled here can explain part of the genetic diversity among breeds and might contribute to explain their origin, history and adaptation to a variety of production systems.

Influence of outdoor rearing and oleic acid supplementation on lipid characteristics of muscle and adipose tissues from obese Alentejano pigs.

Data evaluating the effect of oleic-acid-enriched diets and outdoor rearing (with access to pasture) on the quality of Alentejano (AL) pork and fat are limited. The aim of this work was to study the effects of feeding low (LO) or high oleic acid (HO) diets to AL pigs reared in individual pens (IND) or outdoors (OUT) with access to pasture. Biceps femoris and dorsal subcutaneous fat (DSF) samples were collected to analyse chemical composition and lipids fatty acid (FA) profile. The expression of seven genes involved in lipogenesis, lipolysis and FA transport, as well as two transcription factors were also evaluated in the muscle. B. femoris and DSF presented lower lipid content in OUT-reared than in IND-reared pigs. Two genes involved in lipogenesis, ACACA and FASN, were under-expressed in B. femoris from OUT-reared pigs, while two involved in lipolysis, LPL and HSL were over-expressed. HO diet also significantly reduced FASN expression. Both rearing system and diet affected the FA profile of tissues, particularly DSF. Overall, OUT-reared pigs presented lower n6 to n3 FA ratios in all tissues, and lower monounsaturated (MUFA) and higher polyunsaturated fatty acids (PUFA) content in DSF. When compared to LO-fed ones, HO-fed pigs presented higher oleic acid proportions in all tissues, and lower SFA, higher MUFA and PUFA levels, as well as lower saturation, atherogenic and thrombogenic indexes in DSF. This study shows that rearing system and oleic acid supplementation have complementary effects and influence the nutritional quality of pork and DSF.

Effect of litter birth weight standardization before first suckling on colostrum intake, passive immunization, pre-weaning survival, and growth of the piglets.

Within-litter variation in birth weight is a relevant factor in pig production. This study aimed at comparing pre-weaning mortality, colostrum intake (CI), passive immunization, and growth of piglets from litters of uniform (UN) or heterogeneous (HET) birth weights. The study included 52 multiparous sows (Large White × Landrace) and their litters. Two types of litters were constituted based on birth weight, namely: UN or HET, the control group, using piglets from two to three sows farrowing approximately at the same time. At birth, piglets were weighed, identified, and placed in a box under an IR lamp. At the end of farrowing, piglets were re-weighed and allotted to groups UN or HET (12 per litter) with average weights of 1394 and 1390 g, respectively, and allowed to suckle (time 0). They were re-weighed 24 h later to estimate CI and sows' colostrum yield. At time 0, the average intra-litter CV (%) in weight of experimental litters were 9.3 ±â€¯0.8 (SEM) and 27.8 ±â€¯0.7 in groups UN and HET, respectively (P < 0.001). At 2 days of age, blood samples were taken from the piglets of 11 litters five UN and six HET) and serum Immunoglobulin G (IgG) contents were determined. Mean CI/piglet/litter was similar in both groups, that is, 415 ±â€¯13 in UN and 395 ±â€¯13 g in HET (P = 0.28), but was less variable in UN litters (CV = 22.4 ±â€¯2 vs 36.0 ±â€¯2%, P < 0.001). The IgG levels at 2 days of age were higher in piglets from UN litters (22.5 ±â€¯0.8 vs 18.4 ±â€¯0.7 g/l; P < 0.001) but the CV of IgG levels was not different between litter type (P= 0.46). Mortality up to 21 days of age was lower in UN litters (6.4 vs 11.9%, P = 0.03). The BW at 21 days was not different between litter type (P = 0.25) but it was less variable among piglets from UN litters (CV: 17.1 ±â€¯1.3 vs 25.7 ±â€¯1.3%; P = 0.01). Results reveal that CI is less variable and mortality is lower in piglets from litters of UN birth weight. The results infer that genetic improvement to decrease variation in birth weight within-litter could have a positive effect on homogeneous CI and thus contribute to reducing piglet mortality.

Growth, blood, carcass and meat quality traits from local pig breeds and their crosses.

Contrary to intensive pig production, local pig breeds and their production systems are able to respond to the high criteria and expectations of modern society in regard to some environmental aspects, animal welfare, food quality and healthiness. This study proposes the recovery, study and use of a cross between two local breeds, contributing to animal biodiversity conservation and to the income of local pig producers. This work studied the growth performance and blood, carcass and meat quality traits of Alentejano (AL), Bísaro (BI) and Ribatejano (RI) (AL × BI, BI × AL) castrated male pigs. Raised outdoors, pigs were fed commercial diets ad libitum and killed at ~65 kg (trial 1, n = 10 from each genotype) and ~150 kg BW (trial 2, n = 9 from each genotype). In trial 1, AL and AL × BI attained slaughter weight later than BI and BI × AL pigs, with AL presenting lower average daily gains than the other genotypes (P < 0.001). Alentejano and RI pigs presented higher (P < 0.01) levels of plasma total protein than BI. Overall, carcass traits were affected by genotype, with length (P < 0.01), yield (P = 0.07) and lean cut proportions (P < 0.01) lower in AL than BI, and intermediate values for crossed pigs. Conversely, AL pigs presented higher fat cut proportion (P < 0.01), average backfat thickness (P < 0.001) and 'zwei punkte' fat depth (P < 0.01) than BI and RI pigs. Alentejano pigs also presented higher Longissimus lumborum (LL) intramuscular fat (P < 0.05), myoglobin content and ultimate pH (P < 0.01), but lower total collagen (P < 0.05), drip (P < 0.001) and cooking losses (P < 0.01), and shear force (P < 0.001) than all other genotypes. Finally, LL showed a more intense red colour in AL than in BI pigs. In trial 2, AL pigs confirmed to be a slow-growing obese breed with lower bone and lean cut proportions than BI, and higher LL intramuscular fat, richer colour, lower water loss and higher tenderness. In both trials, RI pigs grew faster, with higher lean and lower fat cut proportions and backfat thickness, and with overall LL characteristics comparable to those observed in AL pigs. This work demonstrates some clear differences between AL and BI breeds while showing that their crosses present intermediate characteristics in most studied traits. These data on RI pigs can be useful to breeders' associations and farmers in order to consider the use of these crosses as an option or complement to pure line breeding.

Genomic diversity, linkage disequilibrium and selection signatures in European local pig breeds assessed with a high density SNP chip.

Genetic characterization of local breeds is essential to preserve their genomic variability, to advance conservation policies and to contribute to their promotion and sustainability. Genomic diversity of twenty European local pig breeds and a small sample of Spanish wild pigs was assessed using high density SNP chips. A total of 992 DNA samples were analyzed with the GeneSeek Genomic Profiler (GGP) 70 K HD porcine genotyping chip. Genotype data was employed to compute genetic diversity, population differentiation and structure, genetic distances, linkage disequilibrium and effective population size. Our results point out several breeds, such as Turopolje, Apulo Calabrese, Casertana, Mora Romagnola and Lithuanian indigenous wattle, having the lowest genetic diversity, supported by low heterozygosity and very small effective population size, demonstrating the need of enhanced conservation strategies. Principal components analysis showed the clustering of the individuals of the same breed, with few breeds being clearly isolated from the rest. Several breeds were partially overlapped, suggesting genetic closeness, which was particularly marked in the case of Iberian and Alentejana breeds. Spanish wild boar was also narrowly related to other western populations, in agreement with recurrent admixture between wild and domestic animals. We also searched across the genome for loci under diversifying selection based on FST outlier tests. Candidate genes that may underlie differences in adaptation to specific environments and productive systems and phenotypic traits were detected in potentially selected genomic regions.

Fat area and lipid droplet morphology of porcine oocytes during in vitro maturation with trans-10, cis-12 conjugated linoleic acid and forskolin.

Lipid droplets (LD) in porcine oocytes form a dark mass reaching almost all cytoplasm. Herein we investigated changes in fat areas, cytoplasmic tone and LD morphology during in vitro maturation (IVM) of porcine oocytes cultured with 100 µM trans-10, cis-12 conjugated linoleic acid (t10,c12 CLA) or 10 µM forskolin at different time periods. Four groups were constituted: control, excipient, t10,c12 CLA and forskolin, with drugs being supplemented during 44 to 48 h and the initial 22 to 24 h in Experiments 1 and 2, respectively. In Experiment 3, forskolin was supplemented for the first 2 h. Matured oocytes were inseminated with frozen-thawed boar semen and cleavage rate recorded. Before and during IVM, samples of oocytes were evaluated for LD, total and fat areas and fat gray value or for meiotic progression. Results showed that forskolin supplementation during 44 to 48 h or 22 to 24 h inhibits oocyte maturation (exp. 1: forskolin = 5.1 ± 8.0%, control = 72.6 ± 5.0%; exp. 2: forskolin = 24.3 ± 7.4%, control = 71.6 ± 5.6%) and cleavage (exp. 1: forskolin = 0.0 ± 0.0%, control = 55.4 ± 4.1%; exp. 2: forskolin = 8.3 ± 3.3%, control = 54.5 ± 3.0%). Forskolin also reduced oocyte and fat areas. In Experiment 3, forskolin negative effect on oocyte maturation and cleavage disappeared, although minor (P ⩽ 0.03) LD and oocyte fat areas were identified at 22 to 24 h of IVM. Oocytes supplemented with t10,c12 CLA during 44 to 48 h presented a lighter (P ⩽ 0.04) colour tone cytoplasm than those of control and forskolin. In conclusion, t10,c12 CLA and forskolin were capable of modifying the distribution and morphology of cytoplasmic LD during porcine oocyte maturation, thus reducing its lipid content in a time-dependent manner.