Tumuli of IA inSouthern Slovenia

MOESAN

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Brittany
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more celtic
Y-DNA haplogroup
R1b - L21/S145*
mtDNA haplogroup
H3c
Is someone here aware of this paper, seemingly recent, and interesting with some Y-R1b-U152? (discovered thank to Bernard SECHER°
Some kind of Italics there?

[h=5]Abstract[/h] DNA analysis demonstrates that all seven individuals buried in an Early Iron Age barrow at Dolge njive, southeast Slovenia, are close biological relatives. Although group composition does not suggest strict adherence to a patrilineal or matrilineal kinship system, the funerary tradition appears highly gendered, with family links through both the male and female line being important in structuring communities. We explore the implications for our understandings of kinship and funerary practices in Early Iron Age southeast Europe.
[h=5]URI[/h] http://hdl.handle.net/10454/19104
[h=5]Version[/h] Accepted manuscript
[h=5]Citation[/h] Armit I, Fischer C-E, Koon H.E.C et al (2022) Kinship practices in Early Iron Age southeast Europe: genetic and isotopic analysis of burials from the Dolge njive barrow cemetery, Dolenjska, Slovenia. Antiquity. Accepted for publication.
 
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Kinship practices in Early Iron Age southeast Europe: genetic and isotopic analysis of
burials from the Dolge njive barrow cemetery, Dolenjska, Slovenia
Abstract
DNA analysis demonstrates that all seven individuals buried in an Early Iron Age barrow at
Dolge njive, southeast Slovenia, are close biological relatives. Although group composition
does not suggest strict adherence to a patrilineal or matrilineal kinship system, the funerary
tradition appears highly gendered, with family links through both the male and female line
being important in structuring communities. We explore the implications for our
understandings of kinship and funerary practices in Early Iron Age southeast Europe.
Key words: ancient DNA; Iron Age; Slovenia; barrows; isotope analysis; kinship
Introduction
The beginning of the Early Iron Age (c. 800–450 BC) in southeast Europe was accompanied
by significant social changes, many of them apparently related to a growing intensity of
contacts and exchange between communities around the head of the Adriatic, and with the
urbanising societies of the wider Mediterranean world. These changes are marked in eastern
Slovenia, as well as in the broader area between the Eastern Alps and Western Pannonia, by
the emergence of new centres of population comprising large hillforts associated with
extensive barrow cemeteries and, in some cases, evidence for iron-working (e.g. Teržan
1990; Mason 1996; Dular & Tecco Hvala 2007; Mason & Mlekuž 2016; Črešnar & Mele
2019; Črešnar et al. 2020). In the Early Iron Age Dolenjska group (southeast Slovenia and
northern Croatia), which is the focus of this paper, funerary rites shifted from cremation
burials in flat cemeteries to inhumation, usually comprising multiple graves under a
substantial earthen barrow, often with significant quantities of grave goods. These new
centres can be linked to the emergence of extended hierarchies that developed to control and
exploit production and inter-regional trade in, for example, iron, salt and amber.
Although it has been suggested that burial in these barrows might have been based on
familial links, with individual barrows being associated with specific lineages (Dular &
Tecco Hvala 2007: 123–6, 237–45; Teržan 2010), this has been hard to demonstrate using
traditional archaeological techniques. As part of the HERA-funded ENTRANS (Encounters
and Transformations in Iron Age Europe) Project (Armit et al. 2014; 2016), osteological and isotope analysis was applied to sites in the region, with further aDNA analysis obtained through the COMMIOS (Communities and Connectivities: Iron Age Britons and their Continental Neighbours) Project. This paper details the results of work on one of these sites,the Dolge njive barrow cemetery, and examines their implications for our widerunderstanding of human mobility and family structure during this dynamic period of southeast European prehistory.The Dolge njive barrow cemeteryThe Dolge njive cemetery forms part of one of the largest mortuary complexes of the Early Iron Age Dolenjska group, which stretches over southeast Slovenia and part of northernCroatia (Figure 1). The complex centres on the large (12.68 ha) hillfort at Veliki Vinji vrh and comprises an estimated 145 barrows. Four main groups ascend to the northwestern entrance of the hillfort from the Topličica valley, while a further 45 dispersed barrows,erected individually or in smaller groups, extend across a wider area of over 25 km2. Many of these barrows were excavated in the late nineteenth century with a relatively poor standard of field-recording and documentation; modern excavation has confirmed, however, that skeletal remains in the area are generally very poorly preserved or absent (Dular & Tecco Hvala 2007: 191; Mason & Mlekuž 2016).The Dolge njive cemetery itself is located between two deeply incised valleys at the foot of the Vinji vrh massif, southeast of the hillfort (Figure 2). Excavations in 2002, in advance of motorway construction, revealed the poorly preserved remains of three Early Iron Age barrows. Two of these were constructed on the site of Late Bronze Age cremation platforms,whilst the third was located a short distance to the east and connected to the others by a Late Bronze Age hollow-way with associated deposits of cremated bone (Mason 2005; Mason & Mlekuž 2016). The remains of an Early Iron Age farmstead or small settlement, comprisingtwo cobbled surfaces and two apparently domestic structures, were discovered at Pod Vovkom to the southwest of the site (Križ 2005; Figure 2). Its location was undoubtedly influenced by the proximity of the Krka River, but may also have taken advantage of one ofthe possible route-ways from the valley to the hillfort.Page 2 of 51

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Two of the Dolge njive barrows (2 and 3) had been largely destroyed by a combination of
Roman settlement activity and medieval agriculture, though both contained at least one
inhumation, in each case accompanied by spearheads (Figure 3a; Table 1). Barrow 1,
however, was better preserved, covering the remains of six graves containing seven
inhumation burials (Figure 3b). All six graves contained extended supine inhumations,
including one double burial (Burial 3) comprising two individuals buried head to toe (Table
1). Five of the graves were arranged in a rough circle around the perimeter of the barrow,
while the other (Burial 1) lay more centrally. This latter grave, however, cut the edge of
Burial 3 and cannot therefore be primary. Although central graves in the region tend to be the
earliest within each barrow, there are exceptions in which they belong to the later or even the
latest phases of a barrow (e.g. Križ 2019: 277, 293).
The limited evidence for inter-cutting in Barrow 1 (Figure 3b), and the slight degree to which
the graves intersect, appears to indicate that the later graves were laid out to respect the
earlier ones: this suggests either that the earlier graves were marked on the surface, and/or
that the graves were dug over a relatively short period. All of the graves contained grave
goods, though the number and composition varied (Table 1); they date on typological
grounds mostly to the Stična (I) phase of the Dolenjska Early Iron Age chronology, i.e. Ha
C(1). Coupled with stratigraphic and aDNA information (see below), the bodies were most
likely deposited over a relatively short period in the early/mid seventh centu
 
Osteological analysis
The skeletal remains were characterised by cortical exfoliation, root etching and were
generally heavily fragmented and incomplete (Nicholls 2017). This was particularly the case
for the less dense bones of the axial skeleton (vertebrae, sterna, ribs and ossa coxae) and
crania, which hampered osteological assessment of age and sex. The dentition exhibited
varying degrees of preservation, but survived most frequently as loose teeth, which permitted
age estimation. All remains appear to belong to young (c. 20–35) and middle adults (c. 36–
50), and we could tentatively assign sex based on skeletal morphology in only five cases
(Table 1). We noted no pathological alterations, although this is unsurprising given the poor
condition of the bones.
 
Ancient DNA analysis
We successfully analysed aDNA from all nine individuals recovered from Dolge njive; seven
from Barrow 1, and single individuals from each of Barrows 2 and 3. The genomic data
obtained for these individuals allowed us to determine genetic sex based on the ratio of Y
chromosome sequences to combined X and Y chromosome sequences (Table 1), as well as
maternal (mitochondrial) and paternal (Y chromosome) lineages (see online Supplementary
Information, which also contains information on the population genetics of the group).
Regarding the mitochondrial lineages, six of the seven samples from Barrow 1 belong to the
H1e5 haplogroup and the seventh carries the H haplogroup. Individuals from Barrows 2 and
3 carry the H5a6 and H1ba haplogroups respectively. All males (across all three barrows)
carry a R1b Y chromosome haplogroup, which is one of the major Y chromosome
haplogroups in Europe following the Late Neolithic/Bronze Age transition; it became
widespread in Europe during the second half of the third millennium BC and is ultimately
linked to ancestry from the Eurasian steppe (Allentoft et al. 2015; Haak et al. 2015).
To explore potential genetic relationships among the Dolge njive individuals, we used the
software READ (Monroy-Kuhn et al. 2015). Within Barrow 1, we found that all seven
individuals were close biological relatives (Figure 4). Burial 5 represents the father of the
individuals in Burials 1, 3a, 3b and 4: three brothers and a sister. The young woman from
Burial 2 is a second-degree relative of these siblings and their father. She is most likely,
therefore, the granddaughter of the man from Burial 5 and niece of the four siblings. Since
she shares her mitochondrial haplogroup with the siblings, it is likely that her mother was a
sister of this group. Burial 6 is a third-degree relative of the siblings (Burials 1, 3a, 3b and 4a)
with whom he shares the same mitochondrial haplogroup. He may be their maternal cousin,
mother’s half-sibling, or great-uncle. Finally, our results highlight that the individuals
interred in Barrows 2 and 3 were not close biological relatives either of each other or of the
family group buried in Barrow 1
 
We analysed multiple isotopes from bones and teeth from all individuals in Barrow 1,
examining evidence for diet and mobility (see Supplementary Information). Where possible,
we sampled multiple elements from each individual to explore intra-individual isotopic
heterogeneity, i.e. lifetime variability. In addition to exploring diet and mobility, the nature of
this assemblage provides a rare opportunity to examine the variation in isotope ratios within a
familial group.
The δ13C and δ15N isotope ratios for the group range from -16.6‰ to -13.6‰ and 7.9‰ to
9.5‰ respectively, indicating a terrestrial-based diet, composed of a mixture of C3 and C4
plants with some herbivorous animal protein (Tykot 2004; Hedges & Reynard 2007). Values
from different elements from each individual show little variation (see Figure 5), indicating
consumption of similar sources of terrestrial protein throughout their lives. δ13CCARB-COL
values consistently exceed 4‰, reflecting a diet high in C4 carbohydrates, likely millet
(Supplementary Table 1; cf. Lightfoot et al. 2013). This is consistent with the limited
previous isotopic analyses for this period in Slovenia (Nicholls et al. 2020; Murray &
Schoeninger 1988) and contemporary botanical evidence (Dular & Tecco-Hvala 2007).
Strontium results are variable, with 87Sr/86Sr ranging from 0.7091 to 0.7102, and
concentrations from 59 to 226 ppm (Supplementary Table 3; Figures 5b and 6). The δ18OCARB
isotope ratios occupy a relatively narrow range of 21.1‰ to 22.8‰. The 86Sr/87Sr values are
consistent with the variable local geology with no evidence for childhood mobility.
Strontium concentrations may reflect the trophic level of food consumed by an individual,
decreasing with increasing trophic level (Evans et al. 2006). The wide variation in strontium
concentrations here (Supplementary Table 3; Figure 6) suggests that two of the brothers
(Burials 3a and 4) consumed more meat and dairy than the rest, while their father (Burial 5),
exhibits by far the highest Sr concentration and thus appears to have eaten more lower trophic
level food. This is consistent with the latter’s δ15N values, which are the lowest in both
dentine and rib collagen. The tooth enamel sampled for this analysis reflects the Sr, O and
CCARB isotopic compositions of food consumed during the formation of the tooth crown in
early childhood. It appears, therefore, that the father (Burial 5) had a different childhood diet
and/or location to his children.
 
AMS dating
Six AMS dates were obtained from the skeletal remains: one from each of the graves in
Barrow 1 (Table 2; Figure 7). The results form a consistent series in the period c. 800–540 cal
BC. The radiocarbon calibration problems associated with the Hallstatt plateau, however, are
such that the AMS dates remain highly imprecise and not readily amenable to further
resolution through Bayesian analysis. They are nonetheless consistent with the typological
dates for the grave goods, which suggest deposition in the first half or the middle of the
seventh century BC.
 
I have jumped over the "discussion" I can post later.
Here some interesting aspect beside the society, kinship and other aspects.



Phenotypes
We used the HIrisPlex SNPs panel (Walsh et al. 2013; Chaitanya et al. 2018), which analyses
41 SNPs, to predict eye and hair colour, and skin pigmentation.
We were able to predict eye colour for four individuals:
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- I5684 has a substantial probability of having blue eyes (76%)
- I5685 has a substantial probability of having blue eyes (64%)
- I5686 has a substantial probability of having blue eyes (76%)
- I5687 has a substantial probability of having blue eyes (78%)
It was not possible to predict hair or skin colour with any confidence due to the absence of
data from too many essential SNPs.
Population Genomics Analysis
In order to carry out population genomics analysis we used the database provided by the
David Reich Laboratory (which curates previously published data from many sources), and
which is available at https://reich.hms.harvard.edu/allen-ancient-dna-resource-aadr-
downloadable-genotypes-present-day-and-ancient-dna-data
The comparative dataset comprises different types of data based on capture and shotgun
sequencing. The shotgun sequencing data are in many cases derived from low coverage
genomes (from 0.1 to 0.5X). This means that the recovered sequences are randomly
distributed across the genome and not targeted as in capture analysis.
Of the first degree relatives, we only use individual I5685 for the following analyses (except
the PCA), as this individual has the best coverage of the group comprising I5685, I5687,
I23971, I22933 and I229
 
I23971, I22933 and I22936 - sorry

Principal Component Analysis
The PCA is built by projecting the ancient DNA data onto a set of modern European and
Near Eastern populations. Here we project available data for Iron Age populations in Western
Europe (Gamba et al. 2014; Allentoft et al. 2015; Schiffels et al. 2016; Martiniano et al.
2016; Damgaard et al. 2018; Mathieson et al. 2018; Antonio et al. 2019; Jarve et al. 2019;
Olalde et al. 2019; Saag et al. 2019; Sikora et al. 2019; Brunel et al. 2020; Fernandes et al.
2020; Marcus et al. 2020; Margaryan et al. 2020).
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Our samples from Dolge njive, as well as others samples from Slovenia, plot in an
intermediate position between Western European samples (UK, France, Spain) and Southern
European samples (Italy) (Figure S1). However, one individual (I22936) seems to have more
affinity with Southern European samples (Italy, Eastern Mediterranean or Punic samples
from Sardinia), but this outlying position potentially reflects the limited coverage (24864
SNPs among the 600 000 SNPs targeted).
Affinities among the Dolge njive samples
We performed a qpWave analysis as implemented in ADMIXTOOLS (Patterson et al. 2012)
to determine if individuals from Dolge njive can be defined as a clade, following Fernandes
et al. 2020. For this test, we used the following set of populations: Mbuti.SDG, EHG,
Russia_Afanasievo, Turkey_N, Germany_EN_LBK, WHG, Russia_Samara_EBA_Yamnaya.
Outliers are defined when p < 0.1.
Our results highlight that all individuals from the Dolge njive site form a clade, including
individual I22936 who is an apparent outlier on the PCA (and who was buried in a separate
barrow from the biologically related individuals in Barrow 1). This is consistent with their
outlying position in the PCA being due to limited data size.
Affinities with ancestral populations
We performed a qpAdm analysis as implemented in ADMIXTOOLS (Haak et al. 2015) to
investigate the percentage of several ancestries in the individuals from Dolge njive. Here we
estimate the relative proportions of EEF, Yamnaya (Steppe) and Western Hunter Gatherer
ancestries. We performed this test on each sample.
Our results indicate that individuals from Dolge njive seem quite homogeneous in terms of
ancestry (Figure S2; note that two individuals (I22933 and I23971) are not plotted as the
model does not fit (p value < 0.05)).
Affinities with contemporaneous populations
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We computed an outgroup-f3 statistic (Raghavan et al. 2014) as implemented in
ADMIXTOOLS in the form (Mbuti; Individual from Dolge njive, Iron Age*) where we
tested the individuals from Dolge njive against the available Iron Age groups. This statistic
allows us to explore the affinities between two groups: a high f3 means that the groups are
close and a low f3 means that the groups show few affinities.
Our results show that the individuals from Dolge njive are closer to other samples from
Slovenia than from other parts of Europe, with the exception of two outliers: one from Spain
and one from England.
Data Availability
The raw data are available as aligned sequences (bam files) through the European Nucleotide
Archive under accession number PRJEBXXXXX [to be completed on acceptance]
 
I cannot post the copy of the PCA, too heavy! Sh...!
 
On PCA, at first sight, the paper studied people are close to other IA Slovenian, at the crossroads of N-Spain, S-France, N-Italy; evoks to me some Italic group or Italo-Etruscan group. Or Illyrians if Illyrians were kind of para-italic speakers (close to Liburnians?) and not close linguistically to Dardanians, closer themselves to proto-Albanians, maybe? Just a bet. No money waited.
 
I cannot post the copy of the PCA, too heavy! Sh...!

Here you go:

x43Hwiu.png


Also, here is the admixture chart.

WIfSj5S.png
 
Thanks, Jovialis, I'm a poor old man of the first steam-computers generation.
 
Has someone some clues about archeologic (cultural) aspect?
 
Thanks, Jovialis, I'm a poor old man of the first steam-computers generation.

No worries, I am some what of a "digital immigrant" myself, since I only had my first computer at age 18. The new generation are true "Digital natives", my toddler already knows how to use a touch screen.
 
It also looks like other IA Italians cluster with the Etruscan/Latins, but also the so-called outliers are present. I'm not sure if this PCA is reusing R437, and R850, but there seems to be a couple more as well. Maybe IA Greeks in Italy? I think the one adjacent to the Scythians, and further to their "west" is the R1 Proto-Villanovan sample.

Totally my speculation:

aLD85Vk.png
 
^^^^^^^^^^^
Are they using different shapes to differentiate among populations or are just using colors?
 
Has someone some clues about archeologic (cultural) aspect?
Dolge njive cemetery tested here is Urnfield classified who were successors of Hügelgräberkultur. Immediate successors of this culture are Hallstatt and further the La Tène Celts.

As was mentioned in the paper: „DNA analysis demonstrates that all seven individuals buried in an Early Iron Age barrow at Dolge njive, southeast Slovenia, are close biological relatives.“ They essentially represent one family/clan. The paternal DNA is R-L2 which is very typical for the before mentioned cultural complex.

These have nothing to do with Cetina/Dinaric culture nor do they match their time frames or archaeogenetic records for that. As far as I can tell they are intermediate between Rhine Celts and IA Latin or IA East Adriatic samples (Cetina/Dinaric) which is to be expected from Cisalpine Celts.
 
Dolge njive cemetery tested here is Urnfield classified who were successors of Hügelgräberkultur. Immediate successors of this culture are Hallstatt and further the La Tène Celts.

As was mentioned in the paper: „DNA analysis demonstrates that all seven individuals buried in an Early Iron Age barrow at Dolge njive, southeast Slovenia, are close biological relatives.“ They essentially represent one family/clan. The paternal DNA is R-L2 which is very typical for the before mentioned cultural complex.

These have nothing to do with Cetina/Dinaric culture nor do they match their time frames or archaeogenetic records for that. As far as I can tell they are intermediate between Rhine Celts and IA Latin or IA East Adriatic samples (Cetina/Dinaric) which is to be expected from Cisalpine Celts.


R-L2 is in NE Italy , istria, croatia as well
 
Dolge Nijve seems to cluster in the range of Cetina culture.

At the autosomal level, yes they show a mean close to the Cetina culture ones, apparently stable since EBA until IA.
That said, they are separated geographically, at least at IA, and the dominant Y haplo in Cetina seems having been Y-J2b-L283.
But the Slovenian tumuli people, Y-R1b all of them, are so close parents that it proves nothing at a less narrow scale of population.
Also, the Cetina ancestors could have come from northern places not too far from Eastern Slovenia.
Only suppositions todate in absence of archeological definitions of the present study tumuli. I 've found nothing on the web helas...
 

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