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Antiquity Vol 80 No 307 March 2006

Indian traders in ancient Bali: a reconsideration of the evidence
Ronald McLauchlan & Richard G. Thomas

Reply
J.S. Lansing, A.J. Redd, T.M. Karafet, J. Watkins, I W. Ardika, S.P.K. Surata, J.W. Schoenfelder, M. Campbell, A.M. Merriwether & M.F. Hammer

Indian traders in ancient Bali: a reconsideration of the evidence

Ronald McLauchlan & Richard G. Thomas

Introduction
Discussions among archaeologists concerning the dates and direction of ancient cultural influences during periods of Southeast Asian prehistory have gradually shifted their emphasis from theories based on mass migration waves to questions concerning smaller groups of people who were apparently able to effect a shift in the material culture of the populations with whom they were in contact. Unfortunately, as yet, no distinctive chemical marker has been identified that will unambiguously indicate nature of these contacts, their scale, their date and the identity of the peoples undertaking these trading activities. In their recent paper An Indian trader in ancient Bali (Lansing et al. 2004), Lansing et al. seemingly broke the impasse, providing both the first earliest direct evidence and a datable marker for individual contact between ancient Bali and India. Since the work presented by Lansing et al. points to a biomarker capable of isolating migrants from the indigenous populations of Southeast Asia, the following study was undertaken in order to assess the wider applicability of this discovery, in particular for our own work on the 'Indianised' culture of Viet Nam. Unfortunately, our results indicate that the scientific evidence for an Indian trader may have been overstated and that much further work is required before the date, forms and directions of 'foreign' influence within Ancient Bali can be isolated and identified.

The argument
The evidence for early trading contact between Bali and India advanced by Lansing et al. rests on four pillars. These are that:

  1. The tooth which forms the focus of the study was found in association with imported objects of Indian origin;
  2. The single radiocarbon determination on the tooth yielded a date consistent with dates for Rouletted Wares and the Kharaoshthi graffito found at the site, both said to be of Indian origin;
  3. Stable carbon isotopic analysis of the tooth yielded a result indicating the individual's primary food source was of terrestrial rather than marine origin (indicative of a 'foreigner' rather than a native Balinese);
  4. Mitochondrial DNA analysis of the tooth combined with computer-based modelling of the Ancient Balinese population, confirmed the foreign extraction and likely north-east Indian ancestry of its original owner.
Unfortunately, on the basis of the evidence published to date, this argument does not seem to be well-founded.

Origins of the sample
The first problem with the proposition advanced by Lansing et al. is that no trace of the tooth or its archaeological context in a trench PCN III is to be found in the publications relating to Ardika's excavations at the site of Pacung. Four fragments of animal bones were recovered from below 3.5m within the single trench opened at Pacung, PCN I, amongst them a tooth of uncertain identification, labelled 'possibly bovine'. If the tooth analysed was indeed that recovered from PCN I then doubts must exist as to the integrity of the archaeological context, the veracity of the date of the sample and the stated association with the Rouletted Ware ceramics. Details of the Pacung site provided by Ardika indicate the tooth was unassociated with any other material indicative of a burial, raising questions as to how the tooth was lost by its original owner. The nature of the stratigraphy within PCN I, makes it impossible to determine whether this tooth was deposited in higher layers, arriving in its final resting place either through natural bioturbidation or via subsequent well digging/infilling operations. It is, for instance, notable that a plastic bag and ballpoint pen were recovered from the in-filled well of PCN I at the same level at which the archaeological material was found within Layer 7!

Dating the sample
Dating the age of the tooth by its association with Rouletted Ware ceramic fragments of Indian origin and glass beads seems doubtful, as only one sherd of Rouletted Ware was found at the PCN I site and no glass beads. The Rouletted Ware, glass beads, the radiocarbon dated pottery sherd and the sherd with Kharoshthi script to which Lansing et al. refer appear to be those recovered from a site at Sembiran, some 300 metres west of Pacung. As noted by the excavators, Pacung and Sembiran are independent sites and were subject to different depositional histories and transformational influences.

Although two human inhumations were indeed discovered in a Sembiran trench (SBN VII), the associations necessary to support the conclusions drawn by Lansing et al. are absent. The human inhumations within SBN VII were not associated with any finds, including Rouletted Wares. Instead, it seems all Rouletted Ware fragments were originally derived from Layer 7 and those found within Layer 6 had been redistributed when the burial pit was cut though into the lower layer. This conclusion seems to be borne out in the results of the AMS radiocarbon dating, which, contra Lansing et al., do not indicate any chronometric relationship between the Rouletted Ware and the inhumations. The dating ranges for two radiocarbon determinations reported for Sembiran indicate that the earliest possible date of manufacture for the Arikamedu type 10 pottery fell within the range 1100 BC to 450 BC. In contrast, AMS radiocarbon dating of the Pacung tooth indicates that the tooth ceased its association with a living person at an indeterminate date somewhere between 340 BC - 20 BC. The only other 14C analysis from Sembiran comes from a sample of unspecified material within Layer 6 located some 0.5 metres above that of the Rouletted Ware. This material ceased actively exchanging carbon with its reservoir at a date somewhere between AD 770 and AD 1240. The dates suggest that the Sembiran burials belong to an entirely separate temporal horizon from that of the Rouletted Wares and throw into question both the chronometric integrity of the Layer 6 and the 14C-date obtained from the tooth (if it is indeed the date of this inhumation that was measured). There is no prima fasciae evidence for any cultural association between a tooth, recovered from either Pacung or Sembiran, and Rouletted Wares. The calibrated date ranges of the Rouletted Wares and the tooth do not overlap and in fact are separated by at least 110 and anywhere up to 1080 years.

Palaeodietary evidence for a 'foreign' tooth
Analytical evidence that the tooth formerly belonged to an individual who was not an indigenous Balinese 'but rather to a foreigner' stems from analysis of the 13C/12C ratio within the tooth. For Lansing et al. (2004) the value of d13C = -21.1‰ indicates that the individual's diet was primarily terrestrial. In contrast, the proximity of the sea to the Sembiran/Pacung sites suggests to the authors that the ancient Balinese inhabitants existed of a diet composed primarily of marine foods, said to be typical of coastal Indonesian peoples.

From the details given of the 13C/12C analyses, such a conclusion would seem to be premature. Any archaeological study based on results obtained from a single tooth and on a comparison with global 13C/12C ranges of various food sources would seem to oversimplify:

  • the dietary choices available to individuals and the possible sources of food;
  • variability of 13C/12C within the local drinking water;
  • the degree of variability of 13C/12C between the different fractions of the tooth structure;
  • spatial and temporal variability of 13C/12C values within species; as well as
  • the degree of post-depositional diagenetic turnover that has taken place.

As it stands, the single figure of d13C = -21.1‰ is meaningless. Its significance should be re-evaluated, after adjustment using appropriate offsets, against the range of possible dietary d13C values for the specified area in Bali and dietary d13C values applicable to 'foreign traders'. As it stands, even if the tooth belonged to a foreigner trader, it is unclear why that person's food intake would have been significantly different to a 'typical' Balinese diet?

Mitochondrial DNA analysis
The study presented by Lansing et al. ultimately focuses upon the results of their study of the mitochondrial DNA within the tooth. Bearing in mind the uncertain origins of the sample and the length of time since excavation, it is regrettable that Lansing et al. provide no information regarding the protocols employed to assess the authenticity of their mtDNA analyses. As the matter stands, their results and conclusions drawn should be regarded with extreme caution.

Several researchers have pointed to the dangers of assaying prehistoric populations using a restricted set of polymorphisms culled from modern populations since these may carry much less genetic variation than their ancestors, or may be more distantly related to ancient populations than is currently recognised. To assay ancient specimens for only a few diagnostic markers is to invite incorrect haplogroup assignments. Furthermore, whilst it has been demonstrated that in cold dry environments mtDNA has the potential to survive in the burial environment for thousands of years these environmental conditions are certainly not those of the Sembiran/Pacung sites where temperature, site hydrology, redox potential, pH and microbial infestation would all be conducive to oxidative and hydrolytic chemical attack and enzymatic attack on the tooth's mtDNA. Post-mortem hydrolytic damage results in deamination and depurination of nucleotide sites in the hypervariable and coding regions of the tooth's mtDNA. This will result in amplified sequences containing artefacts, particularly if a small number of DNA templates initiate the polymerase chain reaction (PCR). These artefacts, which are particularly troublesome if the quantity of pre-PCR mtDNA is low, will, if undetected, lead to incorrect mtDNA typing of the tooth and to subsequent errors in any phylogenetic analysis. In fact, the ancient mtDNA tooth's hypervariable region sites 16261 and those that are associated with haplogroup A (16223, 16290 and 16319) are sites which are susceptible to diagenetic alteration. Given the undoubtedly degraded nature of any existing endogenous tooth mtDNA and that the only reliable indication for a haplogroup A classification is that obtained from the single A-G transition at point 663 in the coding region, any conclusion that the Sembiran tooth necessarily derived from its north-east Indian ancestry would seem to be premature. Even supposing that the mtDNA derived from the tooth is authentic and its mtDNA analysis reliable, the mtDNA haplotype within the 'Pacung tooth' is not unique. Apart from the two private substitutions detected, the haplotype reported for the tooth is present in current Asian populations and is not exclusive to a particular geographic area.

Summary
Unfortunately, too little detail is given regarding the tooth, the circumstances of its discovery and the conditions of its storage to enable anyone to evaluate the validity of the claims now being made for it. On the basis of the currently available information, there is insufficient evidence to substantiate any of the conclusions made for the tooth found at Pacung/Sembiran and in particular the assertion that the tooth belonged to an Indian (foreign) trader present on Bali c. 150-200 BC. It is important to note that this study does not preclude the possibility that the presence of Rouletted Ware and Kharaoshthi graffito may still indicate direct trading links with India. Unfortunately, this conclusion cannot be reached via the evidence presented in the article. It would also seem that major obstacles exist between reliable mtDNA determinations and specifying the provenance of ancient Southeast Asian individuals, at least until some other more distinctive biomarker becomes available.

References


Ronald McLauchlan & Richard G. Thomas: Forensic Sciences, University of Western Sydney, Hawkesbury Campus. Locked Bag 1797, Penrith South DC 1797, Australia.




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Reply

J.S. Lansing, A.J. Redd, T.M. Karafet, J. Watkins, I W. Ardika, S.P.K. Surata, J.W. Schoenfelder, M. Campbell, A.M. Merriwether & M.F. Hammer.

Origins of the sample
As stated in our original report (Lansing et al. 2004), the analysed tooth originated from the excavation unit PCN (Pacung) III, not from PCN I. PCN III was excavated by Ardika and his colleagues in 1999 and 2000 (Tim Jurusan Arkeologi 1999, 2000), after the publication of the English-language articles that were available at the time we submitted to Antiquity.Responding to McLauchlan and Thomas' critique provides us with an opportunity to present some additional information about this work.

PCN III is approximately 210m east of SBN (Sembiran) IX. From the Sembiran 'area of Indian pottery distribution' as presented in Ardika et al. (1997) Figure 1, PCN III is on the far side of a possible former small stream and within the land of the current village of Pacung. PCN III is about 150m north-east of the refilled 1978 well at PCN I, and the well is thus irrelevant to the integrity of the stratigraphic situation in which the tooth was found. The significance of the single sherd of Rouletted Ware (RW) found in 1989 in PCN I has now been confirmed by four RW sherds from PCN II, one from spit 37 of PCN III, and three from spit 38 of PCN III (Tim Jurusan Arkeologi 1999: 44, Ardika 2003: 209-210); further support for a trade connection to India is provided by a base sherd of Arikamedu Type 141 recovered from spit 38 of PCN III (Tim Jurusan Arkeologi 2000: 4; cf. Begley 1991: 182-3).

We did inadvertently describe the sampled tooth as having been discovered 'below spit 3.5'; as McLauchlan and Thomas correctly surmised, we intended to report that it was found 'below 3.5m'. At this depth, between spits 35 and 39 in PCN III, were found not only the four RW sherds from this square, but also our tooth, glass beads, and three flexed human burials, each arranged with its head at the south (toward the mountains). The local pottery wares from these levels are similar to those found with the more numerous Indian sherds at Sembiran, strengthening the impression of general contemporaneity and close cultural relatedness between the Early Metal Period deposits at the Sembiran and Pacung sites (Ardika 2003).

Dating the sample
Our assessment of the age of the sampled tooth is based on the correspondence between the radiocarbon date obtained directly from the tooth itself and the best dates available for the imported ceramics found in the same stratigraphic context. Though each is subject to some uncertainty, the degree of match between the direct date and the contextual date increases our confidence in the general validity of both.

McLauchlan and Thomas write that the direct 14C date obtained from our tooth indicates 'an indeterminate date somewhere between 340 BC - 20 BC'. While not inaccurate, this exaggerates the likelihood of the tooth dating to the third or fourth century BC. For the conventional date received (2110±40 BP; Beta - 161920), the calibrated date range is discontinuous at the two-sigma level as calculated by Beta Analytic at the time of their original reporting, covering both 210 to 40 calBC and 340 to 320 calBC, while excluding years from 320 to 210 BC. Recalibration using the IntCal04 curve and the latest version of the OxCal program (Bronk Ramsey 2001, 2005; Reimer et al. 2004) yields a similar discontinuous range and allows for further clarification: there is a 88.6 per cent probability that the sample dates to between 210 and 30 calBC and a 6.8 per cent probability that it dates to between 350 and 300 calBC.

As established by XRD and NAA studies, the Rouletted Ware found at Sembiran and Pacung is undeniably of South Asian origin (Ardika et al. 1993). This ware travelled widely, and is known from sites in inland and eastern coastal India, from Sri Lanka, and from as far west as the Egyptian Red Sea coast. Based on associations with datable finds including well-known Mediterranean wares, scholars working at different sites have proposed varying initial and final production dates for RW; a range from 250 BC to AD 200 covers most of these estimates (Ardika 1991: 65-66, 70-1; Ardika & Bellwood 1991; Begley 1988: 440, 1991:176-81, 193-4; Coningham & Batt 1999: 128-9; Gogte 1997; Tomber 2000). The director of excavations at Arikamedu, the type site for RW and for other Indian pottery types recovered in association with it at Sembiran and Pacung, considers the Balinese finds indicative of 'contact between the eastern coast of India and Southeast Asia in the first century BC/AD time span (or later)' (Begley 1996: 25).

It is thus entirely plausible that the sampled tooth and RW were deposited at Pacung in the same period, perhaps in or around the first century BC. McLauchlan and Thomas' assertion that 'the calibrated date ranges of the Rouletted Wares and the tooth do not overlap' is simultaneously beside the point and inaccurate: not only does ceramic cross-dating provide a range that does overlap with the tooth's 14C date, but the two Sembiran radiocarbon dates that they reference do not date Rouletted Ware. The later of these two dates (1010±110 BP = 770 - 1260 CalAD via OxCal/IntCal04) is from 50 cm above the Rouletted Ware. As Ardika and Bellwood note, it provides a terminus ante quem (1991: 228). The earlier date (2660±100 BP = 1100 - 400 CalBC via OxCal/IntCal04), which is admittedly problematic (Ardika 1991: 70; Bellwood et al. 1992: 166), was obtained from rice husk temper in a sherd of a large black-slipped storage jar. The ware of this sherd is of probable Indian origin based on XRD and NAA results and has a stratigraphic centre of gravity at Sembiran that suggests its introduction may pre-date RW (Ardika & Bellwood 1991); this increases the plausibility of the arrival of an Indian trader during the earlier portions of the tooth's radiocarbon date range.

It is unclear whether the analysed tooth originally belonged to one of the three human burials recovered at a similar depth in PCN III, but this does not negate the scenario established by ceramic, radiocarbon, and stratigraphic evidence: the tooth was likely deposited in an era in which Indian pottery was also arriving on Bali's shores. Bioturbation may impart a degree of 'fuzziness' to the stratigraphy, but did not completely erase the connection between the tooth and the artefacts. Even if it had, however, we submit that the date itself would be of sufficient interest (given the results of our DNA analyses) to warrant publication for the purposes of encouraging further studies. The question mark in our title indicates the exploratory nature of our original note.

Paleodietary evidence for a 'foreign' tooth
McLaughlan and Thomas suggest that our interpretation of the 13C/12C ratio ignores several sources of case-to-case variability that make citing a single value 'meaningless'. Investigation of the dietary significance of the ratio was originally undertaken to assure ourselves that the effect of 'old' marine reservoir carbon on the apparent age was negligible (Bruce Owen pers. comm. 2002; cf. Barrett et al. 2000). A larger study could have better controlled for potential error sources such as diagenetic change. However, studies of archaeological human remains indicate that d13C values as negative as our sample's (-21.1‰) are quite rare among populations with easy access to marine foods. Since this note is not the place for exhaustive review, we will note only a few examples. In Orkney, Barrett et al. (2000) treated values of -20.6‰ and below as 100 per cent terrestrial diets, and out of 22 samples reported only one with a ratio more negative than ours. In a study in the Marianas, collagen d13C values of -18.1‰ to -15.7‰ were found (n=23), and a calculation based on d13C measurements from local terrestrial animal and C3 plant food resources indicated a hypothetical d13C value of -20.5‰ for a pure C3 diet under prehistoric conditions (Ambrose et al. 1997). A second Marianas study (McGovern-Wilson & Quinn 1996) used a 'most terrestrial' end-point value of -22.53‰, but this was taken from a cow rather than from local flora and fauna; once again, our PCN III individual falls outside the range of observed d13C values for archaeological human remains (-16.6‰ to -19.4‰; n=10).

In general, most researchers agree that there are two main dietary causes for values less negative than ours: diets containing marine food, and diets featuring C4 plants (certain sedges and grasses such as foxtail millet, maize, sugar cane, and sea grasses, though not rice or most other cereals). Disentangling these two factors is impossible without information beyond the d13C value, but this is irrelevant to our particular concern since a d13C of -21.1‰ suggests that neither C4 plants nor sealife made an important contribution to diet. There are also a few diet components that may cause values to shift in a more-negative direction. The 'canopy effect' (Krigbaum 2003), however, primarily affects understory plants and foragers in closed tropical forests. The effects of the consumption of zooplankton-eating seabirds and fish (McGovern-Wilson & Quinn 1996) were insufficient to mask the signature of other marine life in the Marianas, and the same would most likely have been true on Bali. We feel that a parsimonious explanation for the observed ratio is that the Pacung individual was non-local, originating either abroad or from an inland locale on Bali.

Ancient DNA
McLauchlan and Thomas also question the authenticity of our ancient DNA (aDNA) sequence and the reliability of the median-joining network analysis. We list their critiques here along with our responses:

  1. Lansing et al. provide no information regarding the protocols employed. This statement is correct. The aDNA extraction was carried out in a dedicated extraction laboratory that has positive pressure and HEPA filtration. All surfaces are covered by high intensity ultra violet lights. The tooth was split in two with a scratch awl and hammer and the inside was cored out using a dremmel tool inside a custom built aDNA glove box. A phenol-chloroform extraction was carried out using silica during precipitation. The polymerase chain reactions (PCR) were set up in a separate aDNA laboratory using positive and negative controls and a second aDNA glove box. PCR cycling was performed in another building. The extraction and PCR were replicated two times on separate days. None of the individuals who performed the aDNA analysis matched the Bali mtDNA sequence.
  2. Conditions for mtDNA survival are most closely met in cold, dry environments unlike those of the Sembiran/Pacung sites. We agree. Nonetheless, the scientific knowledge of mtDNA preservation is still young and should not preclude the search for ancient mtDNA in teeth, or other biological specimens, from all environments.
  3. The mtDNA might have been damaged via deamination and depurination thus creating sequence artefacts. We think that DNA decay is unlikely. The combination of the restriction site 663G and the control region sites (16189C, 16223T, 16290T, and 16319A) are both consistent with Haplogroup A. Artefacts from DNA decay would not create a consistent fit between the restriction site and the control region substitutions.
  4. Apart from the two private substitutions detected, the haplotype reported for the tooth is present in current Asian populations and is not exclusive to a particular geographic region. This statement is partially correct but misleading. Figure one from Lansing et al. (2004) shows one haplotype with multiple Indian (i.e. Asian) sequences that lack the two private substitutions and that cluster closest to the Bali tooth sequence. With the addition of seven more sequences (see Figure 1) we found that a Mansi (North Asian) individual also joins this set of sequences.. Thus, the tooth haplotype, excluding the two private substitutions, is shared with two Asian populations - five Indians and one Mansi. The weight of the evidence still points toward the subcontinent and not toward the single Mansi individual from North Asia.
  5. A reworking of the Lansing et al. (2004) median network to include additional relevant data is perhaps appropriate. We built the network presented in Figure 1 in response to this suggestion, which was omitted from the final version of McLauchlan and Thomas' comment.

Figure 1

Figure 1. We downloaded 214 global samples from Max Ingman's web site (http://www.genpat.uu.se/mtDB/sequences.php) and searched for haplogroup A sequences and found the following seven individuals: one Canarian (Maca-Meyer et al. 2001); one Mixteca-Baja individual (Mishmar et al. 2003); one Chuckchi (Ingman et al. 2000); one Ket (Starikovskaya et al. 2005); one Mansi (Starikovskaya et al. 2005); one Korean (Mishmar et al. 2003); and one Taiwanese (Mishmar et al. 2003). We built the network with the inclusion of the additional seven haplogroup A sequences (shown in italics). None of these mtDNA sequences matches the ancient tooth sites (663G, 16189C, 16223T, 16240G, 16261T, and 16319A). Click to enlarge.

This network is very similar to the original network. Our conclusions from Lansing et al. (2004) remain the same; we hypothesise that the ancient tooth belonged to an individual of north-east Indian origin.

References

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J.S. Lansing, A.J. Redd, T.M. Karafet, J. Watkins, I W. Ardika, S.P.K. Surata, J.W. Schoenfelder, M. Campbell, A.M. Merriwether & M.F. Hammer.

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