Simone Ceccobelli

Phylogenetic Relationships of Three Italian Merino-Derived Sheep Breeds Evaluated through a Complete Mitogenome Analysis

In Italy, the crisis of the wool industry triggered the necessity to reconvert the two traditional Merino-derived breeds, Gentile di Puglia and Sopravissana, to meat production, by creating the Merinizzata Italiana. The aim of the present study was to assess the genetic diversity of these three Italian Merino-derived (IMd) breeds by examining the molecular information encoded in the maternally-inherited mitochondrial DNA (mtDNA). A parallel molecular investigation was performed on the putative paternal and maternal breeds, the Merino from Spain and the Appenninica from Italy, respectively, as well as on three unrelated dairy breeds (Sarda and Comisana from Italy, and Lacaune from France). Firstly, the mtDNA control region of 291 samples was analyzed. When comparing the overall genetic distances among the eight stocks, the three IMd breeds clustered together close to the Appenninica, thus confirming its parental role. Among the 90 IMd samples, 82 different haplotypes were observed, almost all belonging to haplogroup B, and only one to A. For 23 mtDNAs, including nine IMd, the analysis was then brought to the level of entire mitogenomes. Three distinct sub-haplogroups within B were found to encompass the IMd samples, with one clade (B1a2a1) apparently restricted to those sheep. Thus, despite experiencing a drastic reduction in number (mainly due to changes in breeding practices driven by the economy), the IMd breeds still represent a reservoir of distinctive mitochondrial variants, which could potentially contribute to the development of conservation and management programs of Italian sheep breeds.

Phylogeny, genetic relationships and population structure of five Italian local chicken breeds

Number and population size of local chicken breeds in Italy is considered to be critical. Molecular data can be used to provide reliable insight into the diversity of chicken breeds. The first aim of this study was to investigate the maternal genetic origin of five Italian local chicken breeds (Ancona, Livorno, Modenese, Romagnola and Valdarnese bianca) based on mitochondrial DNA (mtDNA) information. Secondly, the extent of the genetic diversity, population structure and the genetic relationships among these chicken populations, by using 27 microsatellite markers, were assessed. To achieve these targets, a 506 bp fragment of the D-loop region was sequenced in 50 chickens of the five breeds. Eighteen variable sites were observed which defined 12 haplotypes. They were assigned to three clades and two maternal lineages. Results indicated that 90% of the haplotypes are related to clade E, which has been described to originate from the Indian subcontinent. For the microsatellite analysis, 137 individual blood samples from the five Italian breeds were included. A total of 147 alleles were detected at 27 microsatellite loci. The five Italian breeds showed a slightly higher degree of inbreeding (FIS=0.08) than the commercial populations that served as reference. Structure analysis showed a separation of the Italian breeds from the reference populations. A further sub-clustering allowed discriminating among the five different Italian breeds. This research provides insight into population structure, relatedness and variability of the five studied breeds.

A microsatellites-based survey on the genetic structure of two Italian local chicken breeds

The biodiversity safeguard is an important goal of poultry production in every developed country. Nowadays, the high chicken meat demand from the world market has been leading to a large spread of strongly producing commercial chicken lines. The creation of these standard types is causing a progressive loss of genetic variability. Ancona and Livorno are two Italian autochthonous chicken breeds which represent a great resource in terms of specific genetic richness. Aim of this study is to investigate the genetic diversity of these breeds as potential valuable genetic variability source. In fact, in spite of their endangered status, these chicken breeds are very appreciated for their ability to adapt themselves to extensive organic rearing systems. Blood samples from 131 individuals were collected and genotyped through a thirty microsatellitesbased analysis. All the observed descriptive statistical indexes suggested a heterozygosity deficiency and an inbreeding level (mean observed heterozygosity = 0.46, mean expected heterozygosity = 0.53, Fis in Ancona and Livorno = 0.251 and 0.086). The tree from inter-individual DAS distance using Neighbour-Joining algorithm and the FCA analysis showed a higher internal variability in Livorno than in Ancona. STRUCTURE analysis showed the genetic uniqueness of the breeds and the presence of sub-groups in Ancona originating from a possible genetic isolation. This research could be a suitable starting point to set up improved selection schemes and a potential preliminary genotypic test for all the cocks to be used in the selection.

Merino and Merino-derived sheep breeds: a genome-wide intercontinental study

BackgroundMerino and Merino-derived sheep breeds have been widely distributed across the world, both as purebred and admixed populations. They represent an economically and historically important genetic resource which over time has been used as the basis for the development of new breeds. In order to examine the genetic influence of Merino in the context of a global collection of domestic sheep breeds, we analyzed genotype data that were obtained with the OvineSNP50 BeadChip (Illumina) for 671 individuals from 37 populations, including a subset of breeds from the Sheep HapMap dataset.ResultsBased on a multi-dimensional scaling analysis, we highlighted four main clusters in this dataset, which corresponded to wild sheep, mouflon, primitive North European breeds and modern sheep (including Merino), respectively. The neighbor-network analysis further differentiated North-European and Mediterranean domestic breeds, with subclusters of Merino and Merino-derived breeds, other Spanish breeds and other Italian breeds. Model-based clustering, migration analysis and haplotype sharing indicated that genetic exchange occurred between archaic populations and also that a more recent Merino-mediated gene flow to several Merino-derived populations around the world took place. The close relationship between Spanish Merino and other Spanish breeds was consistent with an Iberian origin for the Merino breed, with possible earlier contributions from other Mediterranean stocks. The Merino populations from Australia, New Zealand and China were clearly separated from their European ancestors. We observed a genetic substructuring in the Spanish Merino population, which reflects recent herd management practices.ConclusionsOur data suggest that intensive gene flow, founder effects and geographic isolation are the main factors that determined the genetic makeup of current Merino and Merino-derived breeds. To explain how the current Merino and Merino-derived breeds were obtained, we propose a scenario that includes several consecutive migrations of sheep populations that may serve as working hypotheses for subsequent studies.

Genetic diversity in four populations of Nguni (Zulu) sheep assessed by microsatellite analysis

Zulu sheep are found mainly in the rural KwaZulu-Natal province and the numbers are declining due to indiscriminate inbreeding. There is thus a need for phenotypic and genetic characterisation as a first phase for planning conservation strategies. Zulu sheep populations sampled were from Makhathini research station (MS) (n=33), University of Zululand (UZ) (n=21), a community at KwaMthethwa (KM) (n=32) and from Msinga (EM) (n=33). One European breed Appenninica (AP) was used as out group. Microsatellite analysis using 29 microsatellite loci was used in this study. Among the Zulu sheep, the mean number of alleles per locus was the lowest (3.86) in UZ and the highest (6.24) was realised in EM. The mean values of observed and expected heterozygosity were 0.57 and 0.61, respectively. Neighbour-joining tree showed two main Zulu sheep clusters: the UZ, KM and MS sheep populations clustered together and the second cluster included only representatives from the EM population. The STRUCTURE analysis showed that KM, AP and EM were founded in separate clusters, whereas UZ and MS clustered together. The study demonstrated that there was a common origin of the population from the research stations (MS and UZ populations). It also demonstrated that the EM had a different history for the other three populations. This work suggests that exchange of rams could be useful in reducing inbreeding when considering conservation breeding programmes.

The role of mitochondrial DNA to determine the origin of domestic chicken

Mitochondrial DNA (mtDNA) is has recently lost relevance especially when utilised to study species that are characterised with a history of several migrations. Nonetheless, mtDNA can still represents a useful additional tool in the study of molecular genetic diversity. The reason for the adoption of mtDNA is that it is easy to amplify because it appears in multiple copies in the cells and the mitochondrial gene content is strongly conserved across generations. Thousands of published studies have reached conclusions about population history, patterns of gene flow, genetic structure, and species limits, on the basis of mtDNA sequence variation. MtDNA has been used to study phylo-geographic structure of avian species, and to identify the number of maternal lineages and their geographic origins. Most studies of chicken mtDNA rely on sequences of partial control region but recent researches used the complete mtDNA genome to reconstruct the history of animal domestication. The first genetic study on mtDNA suggests that the Indochinese Red Junglefowl subspecies Gallus gallus gallus is the primary ancestor of the domestic chicken (Gallus gallus domesticus). Other studies showed that at least three subspecies of Gallus gallus were enrolled in the origin of domestic chicken breeds, and that there may be at least two domestication centres: one in Southeast Asia and one in the Indian subcontinent. The authors suggested nine highly divergent clades (named clade A-I) related to geographical distribution in a wide range of domestic chickens and Red Junglefowls across Eurasian regions. Understanding when chickens were transported out of domestication centres and the directions in which they were moved provides information about prehistoric human migration, trade routes and cultural diffusion. MtDNA has been used to infer regions of domestication and to identify the number of maternal lineages and their geographic origins in macroevolution studies.

Genetic diversity and relationships among Italian Merino derived breeds assessed by microsatellites

Abstract The conservation of genetic variability is one of the main objectives in the field of genetics applied to domestic livestock. Among molecular markers, microsatellites are particularly appreciated and widely utilized for the study of animal genome. In this study a genetic characterization of three Italian Merino derived sheep breeds was carried out by 30 microsatellites markers; the genetic relationships between these breeds and the Spanish ancestors were also investigated. All the microsatellites examined resulted polymorphic and a total of 375 alleles were detected. Fis values of the three Italian Merinos demonstrate a low level of inbreeding. The results show a good genetic variability of all the studied breeds; at the same time, the genetic identity of each breed is confirmed. These molecular data can be utilized to improve the present selection schemes and the plans to preserve genetic diversity.

Genetic differentiation between Segugio dell'Appennino and Segugio Maremmano dog breeds assessed by microsatellite markers

The domestic dog (Canis Familiaris) was the first species to be tamed and bred to occupy a variety of functions. The Segugio dell’Appennino and Segugio Maremmano are two Italian breeds used extensively as hunting dogs. Microsatellites and other genetic markers are useful tools in evaluating the within and between breed genetic variation. The aim of this study was to investigate genetic differentiation among the two breeds through the analysis of 21 microsatellite markers. The two populations share the same geographical regions but have been over time selected on different parameters to achieve specific functions. All the analysed microsatellite markers were polymorphic and the average number of alleles per locus was 8.19. The mean FIT index (0.051; P<0.05) highlights that at some point in time, the normal gene flow among the animals was disrupted, giving rise to a heterozygote deficiency in both breeds, and this is confirmed by the mean FST fixation index (0.010; P<0.05) clearly indicating an absence of a significant genetic differentiation between the two breeds. The mean FIS value was significantly different from zero (0.042) (P<0.05) reconfirming the presence of a lack of heterozygosity in the studied samples. The values of observed and expected heterozygosity were similar in the two breeds. AMOVA, PCA and STRUCTURE analysis, all emphasise the lack of significant differences among the two breeds in terms of genetic differentiation. The presence of a population substructure is probably due to a genetic introgression from different Segugio breeds, that can be confirmed with further studies.

Mitochondrial DNA variants of Podolian cattle breeds testify for a dual maternal origin

Background Over the past 15 years, 300 out of 6000 breeds of all farm animal species identified by the Food and Agriculture Organization of the United Nations (FAO) have gone extinct. Among cattle, many Podolian breeds are seriously endangered in various European areas. Podolian cattle include a group of very ancient European breeds, phenotypically close to the aurochs ancestors (Bos primigenius). The aim of the present study was to assess the genetic diversity of Podolian breeds and to reconstruct their origin. Methodology The mitochondrial DNA (mtDNA) control-regions of 18 Podolian breeds have been phylogenetically assessed. Nine non-Podolian breeds have been also included for comparison. Conclusion The overall analysis clearly highlights some peculiarities in the mtDNA gene pool of some Podolian breeds. In particular, a principal component analysis point to a genetic proximity between five breeds (Chianina, Marchigiana, Maremmana, Podolica Italiana and Romagnola) reared in Central Italy and the Turkish Grey. We here propose the suggestive hypothesis of a dual ancestral contribution to the present gene pool of Podolian breeds, one deriving from Eastern European cattle; the other arising from the arrival of Middle Eastern cattle into Central Italy through a different route, perhaps by sea, ferried by Etruscan boats. The historical migration of Podolian cattle from North Eastern Europe towards Italy has not cancelled the mtDNA footprints of this previous ancient migration.

Origin and complete breed standard of Maltese Black breed

This review is a first attempt to give a comprehensive historical account on the evolution of the Maltese Black chicken breed. The initiative consolidates available information to propose hypothesis on the origins of the breed, and to develop a comprehensive breed standard in conformity with the format of the American Breed Standard. The Maltese Black was established in 1950 as a rustic, dual purpose breed capable of producing adequate egg and chicken for consumption. In the 1960s, it was replaced with commercially available stock, marginalising the breed and restricting its existence to small dispersed populations. An initial attempt to have an in-situ conservation of the Maltese Black started in 1998 with a small flock of about 400 chickens that were later relocated to the Agricultural Research and Development Centre in Malta with the intention of maintaining a nucleus flock as a measure for a long term ex-situ conservation strategy. A recent survey highlights that the present population has drifted significantly away from standards first published in 1950. In efforts to consolidate the breed standard definition of the Maltese Black, the breed standards of related Mediterranean breeds, as confirmed by molecular markers, were consulted to translate and address the missing gaps in the previous breed standards. The updated breed standard presented will act as the bench mark against which all future breeding and selection programs are compared.