Discriminant Analysis as a Tool to Classify Grasslands Based on Near-Infrared Spectra.

This study aims to classify plant communities by applying discriminant analysis based on principal components (DAPC) on near-infrared spectra (FT-NIRS) starting from fresh herbage samples. Grassland samples (n~156) belonged to (i) recent alfalfa pure crops (CAA), (ii) recent grass-legume mixtures (GLM), (iii) permanent meadows derived from old alfalfa stands that were re-colonized (PMA), and iv) permanent meadows originated from old grass-legume mixtures (PLM). Samples were scanned using FT-NIRS, and a multivariate exploration of the original spectra was performed using DAPC. The following two scenarios were proposed: (i) cross-validation, where all data were used for model training, and (ii) semi-external validation, where the group assignment was performed without samples of the training set. The first two components explained 98% of the total variability. The DAPC model resulted in an overall assignment success rate of 77%, and, from cross-validation, it emerged that it was possible to assign the CAA and PMA to their group with more than of 80% of success, which were different in botanical and chemical composition. In comparison, GLM and PLM obtained lower success of assignment (~52%). External validation suggested similarity between PLM and GLM groups (93%) and between GLM and PLM (77%). However, a dataset increase could improve group differentiation.

Persistence of autozygosity in crossbreds between autochthonous and cosmopolitan breeds of swine: a simulation study.

Crossbreeding might be a valid strategy to valorize local pig breeds. Crossbreeding should reduce homozygosity and, as a consequence, yield hybrid vigor for fitness and production traits. This study aimed to quantify the persistence of autozygosity in terminal crossbred pigs compared with purebreds and, in turn, identify genomic regions where autozygosity's persistence would not be found. The study was based on genotyping data from 20 European local pig breeds and three cosmopolitan pig breeds used to simulate crossbred offspring. This study consisted of two steps. First, one hundred matings were simulated for each pairwise combination of the 23 considered breeds (for a total of 276 combinations), ignoring the sex of the parent individuals in order to generate purebred and crossbred matings leveraging all the germplasm available. Second, a few preselected terminal-maternal breed pairs were used to mimic a realistic terminal crossbreeding system: (i) Mora Romagnola (boars) or Cinta Senese (boars) crossed with Large White (sows) or Landrace (sows); (ii) Duroc (boars) crossed with Mora Romagnola (sows) or Cinta Senese (sows). Runs of homozygosity was used to estimate genome-wide autozygosity (FROH). Observed FROH was higher in purebreds than in crossbreds, although some crossbred combinations showed higher FROH than other purebred combinations. Among the purebreds, the highest FROH values were observed in Mora Romagnola and Turopolje (0.50 and 0.46, respectively). FROH ranged from 0.04 to 0.16 in the crossbreds Alentejana × Large White and Alentejana × Iberian, respectively. Persistence of autozygosity was found in several genomic segments harboring regions where quantitative trait loci (QTLs) were found in the literature. The regions were enriched in QTLs involved in fatty acid metabolism and associated with performance traits. This simulation shows that autozygosity persists in most breed combinations of terminal crosses. Results suggest that a strategy for crossbreeding is implemented when leveraging autochthonous and cosmopolitan breeds to obtain most of the hybrid vigor.

Discriminant analysis as a tool to classify farm hay in dairy farms.

Hay is one of the primary constituents of ruminant feed, and rapid classification systems of nutritional value are essential. A reliable approach to evaluating hay quality is a combination of visual combined inspection by NIRS analysis. The analysis was carried out on 1,639 samples of hay collected from 2016 to 2021 in northern Italy. Discriminant analysis (DAPC) on five hay types (FOM, forage mixtures; APG, first alfalfa cutting with prevalence of graminaceous >50%; PRA, prevailing alfalfa >50%; PUA, purity alfalfa >95%; and PEM, permanent meadows) was performed by ex-ante visual inspection categorization and NIRS analysis. This study aimed to provide a complementary method to differentiate hay types and classify unknown samples. Two scenarios were used: i) all data were used for model training, and the discriminant functions were extracted based on all samples; ii) the assignment of each group was assessed without samples belonging to the training set group. DAPC model resulted in an overall assignment success rate of 66%; precisely, the success was 84, 79, 69, 37, and 27% for PUA, FOM, PRA, APG, and PEM, respectively. In the second scenario, three groups showed percentages of posterior assignment probability higher than 70% to only one group: PUA with PRA (~ 99%), PRA with PUA (~71%), and PEM with FOM (~75%). Discriminant analysis can be successfully used to differentiate hay types and could also be used to assess factors related to hay quality in addition to NIRS analysis.

Effects of a Ceiling Fan Ventilation System and THI on Young Limousin Bulls' Social Behaviour.

The study investigated the relationship between the temperature humidity index (THI) and the behaviour of 24 young fattening Limousin bulls reared in two farms in Tuscany, Italy. In each farm, six animals were undergone to ceiling fans (switched on at THI values up to 72), and six animals represented the control group. The trial lasted three days for two consecutive weeks in August 2020. Behavioural observations were conducted using scan sampling technique and eating, ruminating, drinking, resting and other social activities were registered every 5 min, from 9.30 am to 4.00 pm. Two different microclimatic conditions were evaluated to assess the effect of the ventilation system: normal (THI < 78) and alert (THI ≥ 78) conditions. Results showed that the ventilation system had significant effects increasing inactivity and lying down compared to control groups and decreasing eating and drinking activities. THI alert condition caused a significant decrease in eating and an increase in lying down behaviours. Ventilation system did not influence the animals' cleanliness. The ceiling fans' efficiency in changing the behaviour of young fattening bulls was demonstrated but further studies are needed to assess the ventilation system effects, especially during longer heat stress periods.

Influence of Wet Ageing on Beef Quality Traits.

Fresh samples of Longissimus thoracis of Charolais (n = 12), Romagnola (n = 15), Limousine (n = 77), and crossbreed (n = 62) animals were evaluated with different storage periods (0, 4, 9, and 14 days). Proximate analysis (i.e., pH, humidity, color, free water content, and physical parameters) was performed for each sample. The data obtained were evaluated with a mixed model, setting 5 fixed effects (breed, storage time, animals age, EUROP conformation, number of animal transports) and the animal as random. The results demonstrated that meat quality was affected mainly by the wet ageing period and that the visual and tactile parameters were also found to be susceptible to the storage time. The conservation entailed a decrease in meat humidity and an increase in L* and b* traits; it also led to a decrease in the hardness of the sample, in turn affecting the other texture profile analysis parameters considered. Fixed effects affected in different ways the traits analyzed i.e., pH and humidity changed with breed, as well as with EUROP classification, animals' age for some TPA parameters, and the number of animal transports for both visual and tactile parameters. Wet ageing influenced the meat quality, often improving it, confirming how important further research would be to identify precise storage times in relation to the parameters studied.

Admixture and breed traceability in European indigenous pig breeds and wild boar using genome-wide SNP data.

Preserving diversity of indigenous pig (Sus scrofa) breeds is a key factor to (i) sustain the pork chain (both at local and global scales) including the production of high-quality branded products, (ii) enrich the animal biobanking and (iii) progress conservation policies. Single nucleotide polymorphism (SNP) chips offer the opportunity for whole-genome comparisons among individuals and breeds. Animals from twenty European local pigs breeds, reared in nine countries (Croatia: Black Slavonian, Turopolje; France: Basque, Gascon; Germany: Schwabisch-Hällisches Schwein; Italy: Apulo Calabrese, Casertana, Cinta Senese, Mora Romagnola, Nero Siciliano, Sarda; Lithuania: Indigenous Wattle, White Old Type; Portugal: Alentejana, Bísara; Serbia: Moravka, Swallow-Bellied Mangalitsa; Slovenia: Krskopolje pig; Spain: Iberian, Majorcan Black), and three commercial breeds (Duroc, Landrace and Large White) were sampled and genotyped with the GeneSeek Genomic Profiler (GGP) 70 K HD porcine genotyping chip. A dataset of 51 Wild Boars from nine countries was also added, summing up to 1186 pigs (~ 49 pigs/breed). The aim was to: (i) investigate individual admixture ancestries and (ii) assess breed traceability via discriminant analysis on principal components (DAPC). Albeit the mosaic of shared ancestries found for Nero Siciliano, Sarda and Moravka, admixture analysis indicated independent evolvement for the rest of the breeds. High prediction accuracy of DAPC mark SNP data as a reliable solution for the traceability of breed-specific pig products.

Keep Garfagnina alive. An integrated study on patterns of homozygosity, genomic inbreeding, admixture and breed traceability of the Italian Garfagnina goat breed.

The objective of this study was to investigate the genetic diversity of the Garfagnina (GRF) goat, a breed that currently risks extinction. For this purpose, 48 goats were genotyped with the Illumina CaprineSNP50 BeadChip and analyzed together with 214 goats belonging to 9 other Italian breeds (~25 goats/breed), whose genotypes were available from the AdaptMap project [Argentata (ARG), Bionda dell'Adamello (BIO), Ciociara Grigia (CCG), Di Teramo (DIT), Garganica (GAR), Girgentana (GGT), Orobica (ORO), Valdostana (VAL) and Valpassiria (VSS)]. Comparative analyses were conducted on i) runs of homozygosity (ROH), ii) admixture ancestries and iii) the accuracy of breed traceability via discriminant analysis on principal components (DAPC) based on cross-validation. ROH analyses was used to assess the genetic diversity of GRF, while admixture and DAPC to evaluate its relationship to the other breeds. For GRF, common ROH (more than 45% in GRF samples) was detected on CHR 12 at, roughly 50.25-50.94Mbp (ARS1 assembly), which spans the CENPJ (centromere protein) and IL17D (interleukin 17D) genes. The same area of common ROH was also present in DIT, while a broader region (~49.25-51.94Mbp) was shared among the ARG, CCG, and GGT. Admixture analysis revealed a small region of common ancestry from GRF shared by BIO, VSS, ARG and CCG breeds. The DAPC model yielded 100% assignment success for GRF. Overall, our results support the identification of GRF as a distinct native Italian goat breed. This work can contribute to planning conservation programmes to save GRF from extinction and will improve the understanding of the socio-agro-economic factors related with the farming of GRF.

Genetic diversity and population history of eight Italian beef cattle breeds using measures of autozygosity.

In the present study, GeneSeek GGP-LDv4 33k single nucleotide polymorphism chip was used to detect runs of homozygosity (ROH) in eight Italian beef cattle breeds, six breeds with distribution limited to Tuscany (Calvana, Mucca Pisana, Pontremolese) or Sardinia (Sarda, Sardo Bruna and Sardo Modicana) and two cosmopolitan breeds (Charolais and Limousine). ROH detection analyses were used to estimate autozygosity and inbreeding and to identify genomic regions with high frequency of ROH, which might reflect selection signatures. Comparative analysis among breeds revealed differences in length and distribution of ROH and inbreeding levels. The Charolais, Limousine, Sarda, and Sardo Bruna breeds were found to have a high frequency of short ROH (~ 15.000); Calvana and Mucca Pisana presented also runs longer than 16 Mbp. The highest level of average genomic inbreeding was observed in Tuscan breeds, around 0.3, while Sardinian and cosmopolitan breeds showed values around 0.2. The population structure and genetic distances were analyzed through principal component and multidimensional scaling analyses, and resulted in a clear separation among the breeds, with clusters related to productive purposes. The frequency of ROH occurrence revealed eight breed-specific genomic regions where genes of potential selective and conservative interest are located (e.g. MYOG, CHI3L1, CHIT1 (BTA16), TIMELESS, APOF, OR10P1, OR6C4, OR2AP1, OR6C2, OR6C68, CACNG2 (BTA5), COL5A2 and COL3A1 (BTA2)). In all breeds, we found the largest proportion of homozygous by descent segments to be those that represent inbreeding events that occurred around 32 generations ago, with Tuscan breeds also having a significant proportion of segments relating to more recent inbreeding.

Estimation of Linkage Disequilibrium and Effective Population Size in Three Italian Autochthonous Beef Breeds.

The objective was to investigate the pattern of linkage disequilibrium (LD) in three local beef breeds, namely, Calvana (n = 174), Mucca Pisana (n = 270), and Pontremolese (n = 44). As a control group, samples of the Italian Limousin breed (n = 100) were used. All cattle were genotyped with the GeneSeek GGP-LDv4 33k SNP chip containing 30,111 SNPs. The genotype quality control for each breed was conducted separately, and SNPs with call rate < 0.95 and minor allele frequency (MAF) > 1% were used for the analysis. LD extent was estimated in PLINK v1.9 using the squared correlation between pairs of loci (r2) across autosomes. Moreover, r2 values were used to calculate historical and contemporary effective population size (Ne) in each breed. Average r2 was similar in Calvana and Mucca Pisana (~0.14) and higher in Pontremolese (0.17); Limousin presented the lowest LD extent (0.07). LD up to 0.11-0.15 was persistent in the local breeds up to 0.75 Mbp, while in Limousin, it showed a more rapid decay. Variation of different LD levels across autosomes was observed in all the breeds. The results demonstrated a rapid decrease in Ne across generations for local breeds, and the contemporary population size observed in the local breeds, ranging from 41.7 in Calvana to 17 in Pontremolese, underlined the demographic alarming situation.

Population Structure and Genetic Diversity of Italian Beef Breeds as a Tool for Planning Conservation and Selection Strategies.

The aim was to investigate the population structure of eight beef breeds: three local Tuscan breeds under extinction, Calvana (CAL), Mucca Pisana (MUP), and Pontremolese (PON); three local unselected breeds reared in Sardinia, Sarda (SAR), Sardo Bruna (SAB), and Sardo Modicana (SAM); and two cosmopolitan breeds, Charolais (CHA) and Limousine (LIM), reared in the same regions. An effective population size ranges between 14.62 (PON) to 39.79 (SAM) in local breeds, 90.29 for CHA, and 135.65 for LIM. The average inbreeding coefficients were higher in Tuscan breeds (7.25%, 5.10%, and 3.64% for MUP, CAL, and PON, respectively) compared to the Sardinian breeds (1.23%, 1.66%, and 1.90% in SAB, SAM, and SAR, respectively), while for CHA and LIM they were <1%. The highest rates of mating between half-siblings were observed for CAL and MUP (~9% and 6.5%, respectively), while the highest rate of parent-offspring mating was ~8% for MUP. Our findings describe the urgent situation of the three Tuscan breeds and support the application of conservation measures and/or the development of breeding programs. Development of breeding strategies is suggested for the Sardinian breeds.