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Paleoparasitological study on the soil sediment samples from archaeological sites of ancient Silla Kingdom in Korean peninsula
Quaternary International xxx (2015) 1e7
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Paleoparasitological study on the soil sediment samples from archaeological sites of ancient Silla Kingdom in Korean peninsula Myeung Ju Kim a, Min Seo b, Chang Seok Oh c, Jong-Yil Chai d, Jinju Lee e, Gab-jin Kim f, Won Young Ma g, Soon Jo Choi h, Karl Reinhard i, Adauto Araujo j, Dong Hoon Shin c, * a
Department of Anatomy, Dankook University College of Medicine, San 29, Anseo-Dong, Cheonan 330-715, South Korea Department of Parasitology, Dankook University College of Medicine, San 29, Anseo-Dong, Cheonan 330-715, South Korea Bioanthropology and Paleopathology Lab, Institute of Forensic Science, Seoul National University College of Medicine, 28 Yongon-Dong, Chongno-Gu, Seoul 110-799, South Korea d Department of Parasitology and Tropical Medicine, Seoul National University College of Medicine, 28 Yongong-Dong, Chongno-Gu, Seoul 110-799, South Korea e Woori Research Institute of Cultural Properties, Changwon, South Korea f Dongyang Research Center of Korean Heritage, Busan Metropolitan City, South Korea g The Korea Institute of Heritage, Hanam-si, South Korea h Foundation of Silla Cultural Heritage Research Institute, South Korea i School of Natural Resources, University of Nebraska-Lincoln, Lincoln, NE 68588, USA j ~o Oswaldo Cruz, FIOCRUZ, Manguinhos, Rio de Janeiro 21040-360, Brazil Fundaça b c
a r t i c l e i n f o
a b s t r a c t
Article history: Available online xxx
Due to paleoparasitology's relatively late beginnings, the fundamental data necessary for any reasonably complete understanding of parasitic infection patterns in Korean history remains insufficient. Especially with respect to ancient samples dating to before the Joseon Dynasty (1392e1910 CE), few cases have been analyzed by parasitological techniques. In the present study, we therefore undertook a series of paleoparasitological examinations of archaeological samples from the ancient Silla Kingdom (57 BCE e935 CE). Specifically, in soil samples obtained from shell midden and mountain top fortress sites, we observed Ascaris lumbricoides, Trichuris trichiura, Dicrocoelid and Taenia sp. eggs. The results of our current investigation of strata and samples from the prosperous first-millennium era of the Silla Kingdom are encouraging for a comprehensive understanding of the parasitic infection patterns in the earlier days of Korean history that could not be obtained by any previous studies. © 2015 Elsevier Ltd and INQUA. All rights reserved.
Keywords: Parasite egg Korea Ancient Paleoparasitological study Silla Kingdom
1. Introduction Paleoparasitology is the scientific study of ancient parasite eggs or larvae remnant in archaeological samples, usually after long burial durations. Paleoparasitologists, in examining soil sediments or coprolite samples obtained from archaeological sites worldwide, discover the evidence of ancient parasitic infections by conventional and/or molecular parasitological techniques. They not only confirm the existence of specific parasitic infections in the past, but also trace the histories of hosteparasite relationships, thereby broadening our knowledge of the evolution of parasitic infections (Ferreira et al., 2000, 2011; Reinhard and Araujo, 2008; Seo et al., 2014a). Paleoparasitological studies have also been conducted in South Korea. Even before full-fledged studies began, pioneering * Corresponding author. E-mail address: [email protected] (D.H. Shin).
researchers had made significant contributions to the establishment of the discipline in that country. The earliest paleoparasitological study in South Korea was carried out on samples from ShinchangDong, a famous prehistoric wetland site (Kanehara and Kanehara, 1997). In another pioneering Korean study on archaeological soil samples, ancient parasite eggs were also detected, specifically Ascaris, Trichuris and Clonorchis eggs, in pit-soil samples from the Unified Silla period (668e935 CE). In so doing, they demonstrated the potential of archaeological sites as fruitful fields of paleopathological endeavor for later investigations (Han et al., 2003). From those early years, paleoparasitological study steadily gained momentum in Korea. Thus far, the most remarkable information has been gleaned from archaeological samples (i.e. mummy coprolites or soil sediments) dating to the Joseon period (1392e1910 CE). In a series of such studies, many questions with respect to historically changing patterns of parasitic infection have begun to be answered (Seo et al., 2007, 2008, 2010, 2011, 2014a,
http://dx.doi.org/10.1016/j.quaint.2015.02.007 1040-6182/© 2015 Elsevier Ltd and INQUA. All rights reserved.
Please cite this article in press as: Kim, M.J., et al., Paleoparasitological study on the soil sediment samples from archaeological sites of ancient Silla Kingdom in Korean peninsula, Quaternary International (2015), http://dx.doi.org/10.1016/j.quaint.2015.02.007
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M.J. Kim et al. / Quaternary International xxx (2015) 1e7
2014b; Lee et al., 2009, 2011; Shin et al., 2009a, 2009b, 2011a, 2011b, 2012a, 2012b, 2013a; Oh et al., 2010a, 2010b). Despite remarkable contributions such as these, however, overall parasitic infection patterns in Korean history have remained obscure. The previous outcomes were mainly obtained from relatively recent Joseon period (1392e1910 CE) samples. From samples dating to earlier historical eras, only scant information has been forthcoming. In order to fill this research lacuna, archaeologists and parasitologists have closely collaborated over the past several years to acquire information on the parasitic infection patterns prevailing in the distant Korean past. As an integral part of that effort, the current study investigated excavation sites of the ancient Silla Kingdom (57 BCE e 935 CE). During the first millennium CE, several tribes and kingdoms in the southeastern part of what is now South Korea had vied for hegemony. Silla (57 BCE e 935 CE) emerged as the overall victor, annexing its adversaries throughout the entire Korean peninsula by 676 CE. As is consistent with the fact that the Silla Kingdom prospered for nearly a thousand years in Gyeongsang province, developing from a small city-state into a well-organized kingdom with a vast territory, many cultural remains have been discovered in the present southeastern provinces of the Korean peninsula, the political and cultural heartland of the kingdom. Paleoparasitological studies on the same provinces will thereby provide us with invaluable chances to study one of the most prosperous areas in the ancient history of Korea. Our series of examinations of samples from archaeological sites dating to the Silla period will afford evidence better enabling us to understand historical Korean parasitic infection patterns more comprehensively. 2. Materials and methods Our paleoparasitological studies in the heartland of the ancient Silla Kingdom were mainly conducted at archaeological sites in the Gyeongsang provinces, the southeastern part of the Korean
peninsula. Archeological excavations at each site were carried out by the Silla Cultural Heritage Research Institute (for GNM_A), the Korea Institute of Heritage (for GNM_B), the Woori Research Institute of Cultural Properties (for Hwawang Sansung), and the Dongyang Research Center of Korean Heritage (for Bongwhang-Dong). The archaeological sampling was a collaborative effort between Dankook University and Seoul National University researchers. The locations of all of the archaeological sites investigated are indicated on the map provided in Fig. 1. In order to move ahead with our ancient parasitology investigation, archaeologists delicately examined the geological strata at each archaeological site. The dates of the soil strata were estimated based on the available cultural and geological information, and most of the dates corresponded to the period of Silla Kingdom. After pinpointing the candidate strata, the parasitologists collected samples from them. To minimize the risk of contamination, soil samples were collected while wearing protective gloves, sterilized gowns, and masks (Fig. S1). Owing to the fact that our sampling was conducted on soils left exposed at open-air sites, surface soils within a 1-m radius of the site also were sampled as negative controls. When the results on the surface soils showed negativity, the data from the geological strata were regarded as authentic. The archaeological information on the soil samples (n ¼ 143) is summarized in Table 1. In Gyeongju City, the Silla capital, we collected samples from two different archaeological sites: GNM_A and GNM_B. The total number of Gyeongju samples was 67. At the GNM_A site, the samples (n ¼ 58) represented four different subareas (A to D). The sampled strata were date-estimated to the 7th to 9th century (the Unified Silla period in Korean history). Three samples (B to D) were collected from the strata that had been in side gutters on the streets of Gyeongju city when it was the Silla capital. Sewage in those streets almost certainly would have been drained through them (Fig. S2). At the GNM_B site, the nine sediment samples collected had been in a storage place dating to the 7th to 9th century CE (Fig. S3).
Table 1 The archaeological information of the Silla samples used in this study. District
Sites/strata in the district
GNM_A (Gyeongju)
A B C D Seokjo Yugu 1
GNM_B (Gyeongju) Bonghwang-Dong (Gimhae)
Hwawang Sansung (Changnyeong)
Sample number 24 5 17 12 9
Estimated period
Archaeological information
Silla 7the9th Century CE
Not determined for its usage in Silla period Side Gutter in the capital of Silla kingdom
A storing place
Joseon Goryeo Flood Precipitates A
6 3 6
Silla 7the9th Century CE Joseon Goryeo ND
Shell-midden Flood Precipitates B I
6 6
Silla Silla
12
Silla Before unification of Korean peninsular in 676 CE
II
14
Silla 6the9th Century CE
III
10
Silla Before unification of Korean peninsular in 676 CE Joseon Silla
IV V Total
5 8 143
The geological layer of Joseon period The geological layer of Goryeo period The geological layer is situated between those of Goryeo and Silla periods. It could be made by flood overflow. The exact date could not be determined by archaeologists because any cultural remains for date estimation were not found in the layer Shell-midden layer of Silla period The geological layer made by flood overflow. Possibly made during Silla period Sample from Grid 4. The geological strata formed at the interim period of Reservoirs #2 and #3. Corresponded to Ancient Silla kingdom stratum before unification of Korean peninsula (676 CE) Sample from precipitates in Reservoir#2 (inside). The geological strata were formed during the reservoir sill in use. Reservior#2 looks to have been mainly used after unification of Korean peninsula Sample from Reservoir#3 (base). The reservoir layer was formed before unification of Korean peninsula Geological strata formed during Joseon period Geological strata formed during Sillla period
Please cite this article in press as: Kim, M.J., et al., Paleoparasitological study on the soil sediment samples from archaeological sites of ancient Silla Kingdom in Korean peninsula, Quaternary International (2015), http://dx.doi.org/10.1016/j.quaint.2015.02.007
M.J. Kim et al. / Quaternary International xxx (2015) 1e7
Fig. 1. The map of South Korea showing sampling sites in this study. Red-shaded area is Gyeongsang provinces where we performed the sampling for the current study. A, Gyeongju; B, Changnyeong; C, Gimhae. We obtained parasite-egg positive results from the various archaeological sites of the ancient Silla period. The current results form a striking contrast to our previous paleoparasitological study on the samples from southwestern part of South Korea (blue-shaded area). The study in 2011, though we examined 593 samples from 17 sites, showed very few positive findings (Kim et al., 2013). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
At the Bongwhang-Dong site, we conducted sampling from five different geological strata (Joseon, Goryeo, Flood_Precipitates_A, Shell-midden, and Flood_Precipitates_B). The total number of samples was 27. Of these, the Shell-midden (n ¼ 6) and
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Flood_Precipitates_B samples (n ¼ 6) corresponded to the Silla period. The Flood_Precipitates_A strata age could not be estimated, due to historical disturbances by floods. For comparative purposes, we also sampled from Joseon (n ¼ 6) and Goryeo (n ¼ 3) strata (Fig. S4). Additionally, we conducted sampling at an ancient fortress built on the top of Hwawang Mountain. The total number of samples collected there was 49. Sample sets I, II and III represented 6th to 9th century ruins of a reservoir that had been constructed and maintained by the Silla people. Samples I and III were deemed likely to have been formed before the unification of the Korean peninsula (676 CE), while sample II was estimated to have been precipitated post-unification (the Unified Silla period: 676e935 CE). Sample V, another Silla sample, was collected from a site very close to the south wall of the fortress. We also obtained Joseon samples (sample IV, n ¼ 5) for comparison (Fig. S5). The detailed sample information is summarized in Table 1. After transporting the samples to our lab, we prepared them for light-microscopic examination (Callen and Cameron, 1960; Criscione et al., 2007). Briefly, the samples were re-hydrated in 0.5% trisodium phosphate solution and filtered through multiplelayer gauze. After one additional precipitation, the upper turbid layer was discarded, and the precipitates were dissolved in 10% neutral-buffered formalin. The precipitate solutions were then dropped onto slides and examined under light microscopy (Olympus, Japan). A total of 200 ml of each sample (10 ml 20 times each or 20 ml 10 times each) was used. The sizes of the eggs were measured, and the numbers of parasite eggs per gram (EPG) was also estimated. 3. Results There were no parasite eggs discovered in the negative-control samples from the surface soils, ruling out any possibility of false
Fig. 2. Results of paleoparasitological examination on Bongwhang-Dong sample. Top figure is geological strata where we sampled. Dots indicate each sampling point. Yellow and red dots indicate the points showing negative and positive results, respectively. The eggs (bottom) are those identified in the samples. Scale bars ¼ 20 mm. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Please cite this article in press as: Kim, M.J., et al., Paleoparasitological study on the soil sediment samples from archaeological sites of ancient Silla Kingdom in Korean peninsula, Quaternary International (2015), http://dx.doi.org/10.1016/j.quaint.2015.02.007
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M.J. Kim et al. / Quaternary International xxx (2015) 1e7
positivity caused by accidental introduction of surface soils into the examined geological strata. In the cases of the samples from GNM_A and _B (Gyeongju) representing the Silla period, we could not identify any ancient parasite eggs, even though they had been collected from highly populated areas of the Silla capital. However, in geological-strata samples from Bongwhang-Dong and Hwawang Sansung, we observed many parasite eggs. At the Bongwhang-Dong site, ancient parasite eggs were found in the samples taken from the Goryeo, Flood_Precipitates_A, Shellmidden and Flood_Precipitates_B layers, whereas none was found in the same site's Joseon layer (Fig. 2). The Shell-midden and Food_Precipitates_B layers showed particularly heavy contamination during the Silla period. With regard to the samples from the Hwawang Sansung site, we found that most of the Silla samples (I, II and III) from the fortress reservoir exhibited ancient parasite eggs. On the other hand, we could not find any eggs in either the Joseon (IV) or Silla sample (V) taken from an area very close to the south wall of the same Silla fortress (Fig. 3). In the samples from the Bongwhang-Dong and Hwawang Sansung sites, numerous Ascaris and Trichuris sp. eggs were identified. Interestingly, in those Bongwhang-Dong samples, dicrocoelid eggs also were discovered (Fig. 2); and in the Hwawang Sansung samples, Taenia sp. eggs, a species very rarely detected in South Korea, were found (Fig. 3). The characteristics of Ascaris, Trichuris, Taenia spp. and dicrocoelid eggs observed in this study are well matched with previous reports on the species (Garcia, 2007; Searcey et al., 2013). The egg sizes and EPGs estimated in this study are summarized in Table 2.
from the pre-20th century strata in the Joseon Dynasty's capital area (a part of what is now modern Seoul Metropolitan city). Our discovery of many eggs prompted a suspicion that pre-modern major cities in East Asia might have been seriously contaminated by parasite eggs that almost certainly have originated from the inhabitants' feces (Shin et al., 2011b, 2013b). The remarkable achievements of paleoparasitological studies on ancient samples notwithstanding, it is still disappointing that most discoveries have been at archaeological sites of relatively recent origin. More in-depth studies on samples representing pre-Joseon periods are therefore needed for a comprehensive and complete understanding of Korea's parasitological history. The results of our current investigation of strata and samples from the prosperous firstmillennium era of the Silla Kingdom are therefore very encouraging. In this study, we found many ancient parasite eggs in the samples from the Silla strata of the Hwawang Sansung (a mountaintop fortress reservoir) and Bongwhang-Dong (shell-midden) sites. The Bongwhang-Dong site was informative because we discovered ancient parasite eggs in shell-midden samples from the Silla period. In Japan, Matsui et al. (2003) observed ancient parasite eggs in samples from a shell-midden site that had been a garbage dump dating to the prehistoric Jomon period. The authors attributed the presence of the eggs there to the shell-midden site's possible role as a kind of toilet in pre-historic Jomon society (Matsui et al., 2003). Our detection of ancient parasite eggs in shell-midden samples from the Silla period thus indicates that the East Asian people still used the midden as a substitute or complementary toilet, or at least, that they might have dumped night soils there, even after the end of the pre-historic age.
Table 2 Positive results of paleoparasitological examinations in this study. Sites
Area/strata in the site
Estimated period
Parasite eggs identified
Average dimensions
Bonghwang-Dong (Gimhae)
Goryeo
Goryeo 935e1392 ND
Trichuris trichiura
51.2 26.6
1.6
Ascaris lumbricoides Trichuris trichiura
55.0 50.0 50.2 22.5
0.8 3.3
Ascaris lumbricoides Trichuris trichiura Ascaris lumbricoides Trichuris trichiura Dicrocoelid Ascaris lumbricoides Trichuris trichiura Taenia spp. Ascaris lumbricoides Trichuris trichiura Trichuris trichiura Ascaris lumbricoides Taenia spp.
61.5 46.0 47.4 25.6 60.0 47.5 50.3 26.9 41.7 26.0 62.4 46.3 49.0 25.2 39.8 38.6 58.3 50.0 47.5 24.5 50.0 25.0 60.0 43.9 35.0 31.0
5.0 0.8 3.3 11.6 2.5 5.0 5.6 0.4 3.2 5.4 2.5 7.0 0.5
Flood Precipitates A Shell-midden
Hwawang Sansung (Changnyeong)
Silla
Flood Precipitates B I
Silla
II
Silla 6the9th Century CE Silla Before unification of Korean peninsular in 676 CE
III
Silla Before unification of Korean peninsular in 676 CE
4. Discussion In general, heavy parasite-egg contamination of soil is known to correlate well with higher parasitic infection prevalence among inhabitants of the given area (Schulz and Kroeger, 1992; Uga et al., 1995, 1997; Chongsuvivatwong et al., 1999). This is no less true of ancient samples from hundred-to-thousand year-old archaeological sites. It is highly probable that detection of parasite eggs in soil samples from ancient or medieval towns or cities will correlate very closely with the parasitic infection patterns among the relevant historical peoples (Monckton, 1995; Matsui et al., 2003; Fernandes et al., 2005; da Rocha et al., 2006; Mitchell et al., 2008). We have experienced such situations in our studies on ancient samples obtained from a variety of archaeological sites in South Korea. One case involved the examination of soil-sediment samples
EPG
Despite the contributions of the studies just noted, information on shell-midden sites in East Asian history, from the perspective of paleopathology at least, remains largely incomplete. We still do not have any idea of when the use of shell-midden sites finally disappeared in East Asian countries. Did the transition coincide with the adoption of more advanced forms of toilets? Only future studies could answer such a question. Next, we discuss the outcomes of the examinations of the Hwawang-Sansung fortress samples. In ancient Korea, many fortresses were built, in every corner of the country and especially on mountaintop areas. These were refuges in times of war, though recent historical studies have revealed that they were more than that as well. Mountain fortresses were established political centers that were significantly populated even in peacetime (Seo, 2010, 2012). Considering the semi-permanent statuses of such
Please cite this article in press as: Kim, M.J., et al., Paleoparasitological study on the soil sediment samples from archaeological sites of ancient Silla Kingdom in Korean peninsula, Quaternary International (2015), http://dx.doi.org/10.1016/j.quaint.2015.02.007
M.J. Kim et al. / Quaternary International xxx (2015) 1e7
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Fig. 3. Results of paleoparasitological examination on Hwawang Sansung samples. The figures for archeological sites represent each area (I to V) where we sampled. Yellow and red dots indicate the sampling points showing negative and positive results, respectively. SW, south wall of the fortress. The bottom figures of parasite eggs are those found in the ancient samples I, II and III. Scale bars ¼ 10 mm. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
fortresses, their inner areas must have been to some extents contaminated by human wastes, and therefore, at least relatively unsanitary. The evidence gathered in the current study supported this speculation. For example, in the fortress reservoir samples dating to the Silla period, we detected ancient parasite eggs. As to the question of why parasite eggs were plentifully identified in the ancient reservoir samples, previous research on ancient moats around permanent settlements can be very informative. For instance, upon our detection of ancient parasite eggs in the samples from a moat constructed around the Silla King's palace in Gyeongju city, we conjectured that the phenomenon might have been caused by a continual feces-contaminated inflow, from the King's palace into the surrounding moat area (Shin et al., 2009c). Inflow-contamination of an ancient moat encircling a permanent settlement also has been reported in Japan. Matsui et al. (2003) found evidence that wastewater or excrement from a settlement dating to the Yayoi period (450 BCEe300 CE) had been continuously discharged into the surrounding moat. Although the stagnant water in the moat might have made for serious sanitary
problems, this system seemingly was indispensable to them in any case, as human waste had to be removed from the settlement somehow. Like the discharging inflows to ancient moats around the settlements (Matsui et al., 2003; Shin et al., 2009c), creeks or streams in the fortress must have flown into the reservoir, which had been constructed at a lower altitude. Human wastes scattered in the fortress and riding on the stream flows could also have been drained into the reservoir. The ancient eggs finally detected in the reservoir samples could well have originated in the feces. Based on our study, we speculated that parasite re-infection must have been highly prevalent among the fortress-dwellers; and the eggs laid by the adult parasites must have been defecated into the soil of the region, contaminating the fortress area continuously during the ancient Silla period. Of the ancient parasite eggs discovered in various Silla samples of this study, dicrocoelid and Taenia eggs have been reported only very rarely in previous works on ancient samples in South Korea (Lee et al., 2011). Dicrocoelium sp. has attracted
Please cite this article in press as: Kim, M.J., et al., Paleoparasitological study on the soil sediment samples from archaeological sites of ancient Silla Kingdom in Korean peninsula, Quaternary International (2015), http://dx.doi.org/10.1016/j.quaint.2015.02.007
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M.J. Kim et al. / Quaternary International xxx (2015) 1e7
paleoparasitologists' interest for its importance to understanding of historical human migrations that have occurred between continents. The detection of dicrocoelid eggs in Western Europe was reported in samples dating to as early as 550,000 years BCE to the 16th century CE, while New World samples began to exhibit the same species at around the 17th century CE, after European colonization (Le Bailly and Bouchet, 2010). However, the number of relevant reports is still not sufficient to constitute a global overview of ancient dicrocoeliosis history. The reports on the presence of ancient Dicrocoelium eggs in European, North American and African archaeological samples have been sporadic (Dittmar and Steyn, 2004). In Asia, no reports on ancient Dicrocoelium eggs have been available until now. In this regard, our detection of Dicrocoelium eggs in ancient Silla samples, the firstever such report for Asia, is very meaningful. At least, this clearly shows that, like any other Old World peoples, the dicrocoelid also infected Asians even before the Columbian discovery of America. Another noteworthy finding in the present study was the detection of Taenia eggs in the Hwawang Sansung fortress samples. According to Matsui et al. (2003), Taenia sp. has been very rarely detected in ancient Japanese samples. An exception was the sample from the cesspits at the Koro-Kan site, the 8th century guesthouse in Japan. About the presence of ancient Taenia sp. eggs in the cesspit samples, Matsui et al. (2003) speculated that the eggs might not have originated from Japanese, but rather, that they could have been deposited by diplomatic delegates from foreign countries. The authors' hypothesis was clearly based upon the speculation that the people in neighboring countries, namely China or Korea, might have been more vulnerable to taeniasis by enjoying pork and beef much freely than could the ancient Japanese. In Japan during the ancient period, meat-eating was strictly prohibited for religious reasons (Matsui et al., 2003). However, Taenia sp. eggs from ancient East Asian samples have only rarely been reported. Among the multiple studies and many samples of the past several years, only one sample, taken from a Joseon tomb, contained Taenia sp. eggs (Lee et al., 2011). Moreover, given the lack of any reports on the presence of ancient Taenia eggs in South Korean samples similar in age to those in the Koro-Kan case, Matsui et al. (2003)'s speculation that the Taenia sp. eggs detected in ancient Japanese samples had actually originated from visiting Korean or Chinese delegates was a subject for further studies. In this regard, the detection of Taenia sp. eggs in the Silla samples is very interesting. According to the current results, it could not be completely ruled out that Taenia sp. infection was prevalent among Korean populations even in ancient times. However, in any case, the hypothesis requires further corroboration in forthcoming research. For instance, the difference in prevalence between ancient Korean and Japanese peoples must also be clarified by future paleoparasitological studies. 5. Conclusions Given the late start of paleoparasitological studies in Korea, fundamental data necessary for any reasonably complete understanding of parasitic infection patterns are still insufficient. Our present series of paleoparasitological examinations of ancient Korean archaeological samples hopefully will go some way toward supplementing that data. In our study, we analyzed soil-sediment samples obtained from sites dating to the Silla Kingdom (57 BCEe935 CE). Specifically, Ascaris lumbricoides, Trichuris trichiura, Taenia spp. and dicrocoelid eggs were observed in a shellmidden and at a mountaintop fortress. We were able to derive invaluable data on parasitic infection patterns of ancient Koreans, information that previously was difficult to obtain.
Acknowledgements The first two authors contributed equally to this study. This study was supported by the National Research Institute of Cultural Heritage, Korea (NRICH-1207-B04F-1). Appendix A. Supplementary data Supplementary data related to this article can be found at http:// dx.doi.org/10.1016/j.quaint.2015.02.007. References Callen, E.O., Cameron, T.W.M., 1960. A prehistoric diet as revealed in coprolites. New Scientist 8, 35e40. Chongsuvivatwong, V., Uga, S., Nagnaen, W., 1999. Soil contamination and infections by soil-transmitted helminths in an endemic village in southern Thailand. The Southeast Asian Journal of Tropical Medicine and Public Health 30, 64e67. Criscione, C.D., Anderson, J.D., Sudimack, D., Peng, W., Jha, B., Williams-Blangero, S., Anderson, T.J., 2007. Disentangling hybridization and host colonization in parasitic roundworms of humans and pigs. Proceedings of the Royal Society B: Biological Sciences 274, 2669e2677. da Rocha, G.C., Harter-Lailheugue, S., Le Bailly, M., Araújo, A., Ferreira, L.F., da SerraFreire, N.M., Bouchet, F., 2006. Paleoparasitological remains revealed by seven rias do Instituto historic contexts from “Place d'Armes”, Namur, Belgium. Memo Oswaldo Cruz 101 (Suppl. 2), 43e52. Dittmar, K., Steyn, M., 2004. Paleoparasitological analysis of coprolites from K2, an Iron Age archaeological site in South Africa: the first finding of Dicrocoelium sp. eggs. Journal of Parasitology 90 (1), 171e173. Fernandes, A., Ferreira, L.F., Gonçalves, M.L., Bouchet, F., Klein, C.H., Iguchi, T., Sianto, L., Araujo, A., 2005. Intestinal parasite analysis in organic sediments collected from a 16th-century Belgian archeological site. Cadernos de saúde pública 21, 329e332. Ferreira, L.F., Britto, C., Cardoso, M.A., Fernandez, O., Reinhard, K., Araújo, A., 2000. Paleoparasitology of Chagas disease revealed by infected tissues from Chilean mummies. Acta Tropica 75, 79e84. Ferreira, L.F., Reinhard, K., Araújo, A., 2011. Fundamentos da Paleoparasitologia. Editora Fiocruz, Rio de Janeiro, p. 484. Garcia, L.S., 2007. Diagnostic Medical Parasitology, fifth ed. ASM Press, Washington D.C. Han, E.T., Guk, S.M., Kim, J.L., Jeong, H.J., Kim, S.N., Chai, J.Y., 2003. Detection of parasite eggs from archaeological excavations in the Republic of Korea. rias do Instituto Oswaldo Cruz 98 (Suppl. l), 123e126. Memo Kanehara, M., Kanehara, M., 1997. Shinchangdong Jeosupji Yujeokeui Hwabunbunseok mit Gisaengchungraneui Bunseok. In: Gwangju Shinchangdong Jeosupji Yujeok, vol. I. National Museum of Gwangju, Gwangju. Kim, M.J., Shin, D.H., Song, M.J., Song, H.Y., Seo, M., 2013. Paleoparasitological surveys for detection of helminth eggs in archaeological sites of Jeolla-do and Jejudo. The Korean Journal of Parasitology 51 (4), 489e492. Le Bailly, M., Bouchet, F., 2010. Ancient dicrocoeliosis: occurrence, distribution and migration. Acta Tropica 115 (3), 175e180. Lee, H.J., Shin, D.H., Seo, M., 2011. Discovery of taeniid eggs from a 17th century tomb in Korea. The Korean Journal of Parasitology 49 (3), 327e329. Lee, I.S., Lee, E.J., Park, J.B., Baek, S.H., Oh, C.S., Lee, S.D., Kim, Y.S., Bok, G.D., Hong, J.W., Lim, D.S., Shin, M.H., Seo, M., Shin, D.H., 2009. Acute traumatic death of a 17th century general based on examination of mummified remains found in Korea. Annals of Anatomy 191, 309e320. Matsui, A., Kanehara, M., Kanehara, M., 2003. Palaeoparasitology in Japanediscovrias do Instituto Oswaldo Cruz 98 (Suppl. 1), ery of toilet features. Memo 127e136. Mitchell, P.D., Stern, E., Tepper, Y., 2008. Dysentery in the crusader kingdom of Jerusalem: and ELISA analysis of two medieval latrines in the City of Acre (Israel). Journal of Archaeological Science 35, 1849e1853. Monckton, A., 1995. Environmental Archaeology in Leicestershire. Transactions of the Leicestershire. Archaeological and Historical Society 69, 32e41. Oh, C.S., Seo, M., Chai, J.Y., Lee, S.J., Kim, M.J., Park, J.B., Shin, D.H., 2010a. Amplification and sequencing of Trichuris trichiura ancient DNA extracted from archaeological sediments. Journal of Archaeological Science 37 (6), 1269e1273. Oh, C.S., Seo, M., Lim, N.J., Lee, S.J., Lee, E.J., Lee, S.D., Shin, D.H., 2010b. Paleoparasitological report on Ascaris aDNA from an ancient East Asian sample. rias do Instituto Oswaldo Cruz 105 (2), 225e228. Memo Reinhard, K.J., Araujo, A., 2008. Archaeoparasitology. In: Pearsall, D.M. (Ed.), Encyclopedia of Archaeology. Academic Press, New York, pp. 494e501. Schulz, S., Kroeger, A., 1992. Soil contamination with Ascaris lumbricoides eggs as an indicator of environmental hygiene in urban areas of north-east Brazil. Journal of Tropical Medicine and Hygiene 95, 95e103. Searcey, N., Reinhard, K.J., Egarter-Vigl, E., Maixner, F., Piombino-Mascali, D., Zink, A.R., van der Sanden, W.A.B., Gardner, S.L., Bianucci, R., 2013. Parasitism of the Zweeloo Woman: Dicrocoeliasis evidenced in a Roman Period Bog Mummy. International Journal of Paleopathology 3 (3), 224e228.
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ancient parasite eggs recovered from Korean mummies of the Joseon Dynasty. Journal of Parasitology 95, 137e145. Shin, D.H., Oh, C.S., Chai, J.-Y., Lee, H.-J., Seo, M., 2011a. Enterobius vermicularis eggs discovered in coprolites from a Medieval Korean Mummy. The Korean Journal of Parasitology 49 (3), 323e326. Shin, D.H., Oh, C.S., Chai, J.Y., Ji, M.J., Lee, H.J., Seo, M., 2012b. Sixteenth century Gymnophalodes seoi infection on the coast of the Korean peninsula. Journal of Parasitology 98 (6), 1283e1286. Shin, D.H., Oh, C.S., Chung, T., Yi, Y.S., Chai, J.Y., Seo, M., 2009c. Detection of parasite eggs from a moat encircling the royal palace of Silla, the ancient Korean Kingdom. Journal of Archaeological Science 36 (11), 2534e2539. Shin, D.H., Oh, C.S., Lee, H.J., Chai, J.Y., Lee, S.J., Hong, D.-W., Lee, S.D., Seo, M., 2013a. Ancient DNA analysis on Clonorchis sinensis eggs remained in samples from medieval Korean mummy. Journal of Archaeological Science 40, 211e216. Shin, D.H., Oh, C.S., Lee, S.J., Chai, J.Y., Kim, J., Lee, S.D., Park, J.B., Choi, I.-H., Lee, H.J., Seo, M., 2011b. Paleo-parasitological study on the soils collected from archaeological sites in old district of Seoul City. Journal of Archaeological Science 38, 3555e3559. Shin, D.H., Oh, C.S., Lee, S.J., Lee, E.-J., Yim, S.G., Kim, M.J., Kim, Y.-S., Lee, S.D., Lee, Y.S., Seo, M., 2012a. Ectopic paragonimiasis from 400 year old female mummy of Korea. Journal of Archaeological Science 39, 1103e1110. Shin, D.H., Oh, C.S., Shin, Y.M., Cho, C.W., Seo, M., 2013b. The pattern of ancient parasite egg contamination in the private residence, alley, ditch and streambed soils of old Seoul City, the capital of Joseon dynasty. International Journal of Paleopathology 3, 208e213. Uga, S., Nagnaen, W., Chongsuvivatwong, V., 1997. Contamination of soil with parasite eggs and oocysts in southern Thailand. The Southeast Asian Journal of Tropical Medicine and Public Health 28 (Suppl. 3), 14e17. Uga, S., Ono, K., Kataoka, N., Safriah, A., Tantular, I.S., Dachlan, Y.P., Ranuh, I.G., 1995. Contamination of soil with parasite eggs in Surabaya, Indonesia. The Southeast Asian Journal of Tropical Medicine and Public Health 26, 730e734.
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Paleoparasitological study on the soil sediment samples from archaeological sites of ancient Silla Kingdom in Korean peninsula Myeung Ju Kim a, Min Seo b, Chang Seok Oh c, Jong-Yil Chai d, Jinju Lee e, Gab-jin Kim f, Won Young Ma g, Soon Jo Choi h, Karl Reinhard i, Adauto Araujo j, Dong Hoon Shin c, * a
Department of Anatomy, Dankook University College of Medicine, San 29, Anseo-Dong, Cheonan 330-715, South Korea Department of Parasitology, Dankook University College of Medicine, San 29, Anseo-Dong, Cheonan 330-715, South Korea Bioanthropology and Paleopathology Lab, Institute of Forensic Science, Seoul National University College of Medicine, 28 Yongon-Dong, Chongno-Gu, Seoul 110-799, South Korea d Department of Parasitology and Tropical Medicine, Seoul National University College of Medicine, 28 Yongong-Dong, Chongno-Gu, Seoul 110-799, South Korea e Woori Research Institute of Cultural Properties, Changwon, South Korea f Dongyang Research Center of Korean Heritage, Busan Metropolitan City, South Korea g The Korea Institute of Heritage, Hanam-si, South Korea h Foundation of Silla Cultural Heritage Research Institute, South Korea i School of Natural Resources, University of Nebraska-Lincoln, Lincoln, NE 68588, USA j ~o Oswaldo Cruz, FIOCRUZ, Manguinhos, Rio de Janeiro 21040-360, Brazil Fundaça b c
a r t i c l e i n f o
a b s t r a c t
Article history: Available online xxx
Due to paleoparasitology's relatively late beginnings, the fundamental data necessary for any reasonably complete understanding of parasitic infection patterns in Korean history remains insufficient. Especially with respect to ancient samples dating to before the Joseon Dynasty (1392e1910 CE), few cases have been analyzed by parasitological techniques. In the present study, we therefore undertook a series of paleoparasitological examinations of archaeological samples from the ancient Silla Kingdom (57 BCE e935 CE). Specifically, in soil samples obtained from shell midden and mountain top fortress sites, we observed Ascaris lumbricoides, Trichuris trichiura, Dicrocoelid and Taenia sp. eggs. The results of our current investigation of strata and samples from the prosperous first-millennium era of the Silla Kingdom are encouraging for a comprehensive understanding of the parasitic infection patterns in the earlier days of Korean history that could not be obtained by any previous studies. © 2015 Elsevier Ltd and INQUA. All rights reserved.
Keywords: Parasite egg Korea Ancient Paleoparasitological study Silla Kingdom
1. Introduction Paleoparasitology is the scientific study of ancient parasite eggs or larvae remnant in archaeological samples, usually after long burial durations. Paleoparasitologists, in examining soil sediments or coprolite samples obtained from archaeological sites worldwide, discover the evidence of ancient parasitic infections by conventional and/or molecular parasitological techniques. They not only confirm the existence of specific parasitic infections in the past, but also trace the histories of hosteparasite relationships, thereby broadening our knowledge of the evolution of parasitic infections (Ferreira et al., 2000, 2011; Reinhard and Araujo, 2008; Seo et al., 2014a). Paleoparasitological studies have also been conducted in South Korea. Even before full-fledged studies began, pioneering * Corresponding author. E-mail address: [email protected] (D.H. Shin).
researchers had made significant contributions to the establishment of the discipline in that country. The earliest paleoparasitological study in South Korea was carried out on samples from ShinchangDong, a famous prehistoric wetland site (Kanehara and Kanehara, 1997). In another pioneering Korean study on archaeological soil samples, ancient parasite eggs were also detected, specifically Ascaris, Trichuris and Clonorchis eggs, in pit-soil samples from the Unified Silla period (668e935 CE). In so doing, they demonstrated the potential of archaeological sites as fruitful fields of paleopathological endeavor for later investigations (Han et al., 2003). From those early years, paleoparasitological study steadily gained momentum in Korea. Thus far, the most remarkable information has been gleaned from archaeological samples (i.e. mummy coprolites or soil sediments) dating to the Joseon period (1392e1910 CE). In a series of such studies, many questions with respect to historically changing patterns of parasitic infection have begun to be answered (Seo et al., 2007, 2008, 2010, 2011, 2014a,
http://dx.doi.org/10.1016/j.quaint.2015.02.007 1040-6182/© 2015 Elsevier Ltd and INQUA. All rights reserved.
Please cite this article in press as: Kim, M.J., et al., Paleoparasitological study on the soil sediment samples from archaeological sites of ancient Silla Kingdom in Korean peninsula, Quaternary International (2015), http://dx.doi.org/10.1016/j.quaint.2015.02.007
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M.J. Kim et al. / Quaternary International xxx (2015) 1e7
2014b; Lee et al., 2009, 2011; Shin et al., 2009a, 2009b, 2011a, 2011b, 2012a, 2012b, 2013a; Oh et al., 2010a, 2010b). Despite remarkable contributions such as these, however, overall parasitic infection patterns in Korean history have remained obscure. The previous outcomes were mainly obtained from relatively recent Joseon period (1392e1910 CE) samples. From samples dating to earlier historical eras, only scant information has been forthcoming. In order to fill this research lacuna, archaeologists and parasitologists have closely collaborated over the past several years to acquire information on the parasitic infection patterns prevailing in the distant Korean past. As an integral part of that effort, the current study investigated excavation sites of the ancient Silla Kingdom (57 BCE e 935 CE). During the first millennium CE, several tribes and kingdoms in the southeastern part of what is now South Korea had vied for hegemony. Silla (57 BCE e 935 CE) emerged as the overall victor, annexing its adversaries throughout the entire Korean peninsula by 676 CE. As is consistent with the fact that the Silla Kingdom prospered for nearly a thousand years in Gyeongsang province, developing from a small city-state into a well-organized kingdom with a vast territory, many cultural remains have been discovered in the present southeastern provinces of the Korean peninsula, the political and cultural heartland of the kingdom. Paleoparasitological studies on the same provinces will thereby provide us with invaluable chances to study one of the most prosperous areas in the ancient history of Korea. Our series of examinations of samples from archaeological sites dating to the Silla period will afford evidence better enabling us to understand historical Korean parasitic infection patterns more comprehensively. 2. Materials and methods Our paleoparasitological studies in the heartland of the ancient Silla Kingdom were mainly conducted at archaeological sites in the Gyeongsang provinces, the southeastern part of the Korean
peninsula. Archeological excavations at each site were carried out by the Silla Cultural Heritage Research Institute (for GNM_A), the Korea Institute of Heritage (for GNM_B), the Woori Research Institute of Cultural Properties (for Hwawang Sansung), and the Dongyang Research Center of Korean Heritage (for Bongwhang-Dong). The archaeological sampling was a collaborative effort between Dankook University and Seoul National University researchers. The locations of all of the archaeological sites investigated are indicated on the map provided in Fig. 1. In order to move ahead with our ancient parasitology investigation, archaeologists delicately examined the geological strata at each archaeological site. The dates of the soil strata were estimated based on the available cultural and geological information, and most of the dates corresponded to the period of Silla Kingdom. After pinpointing the candidate strata, the parasitologists collected samples from them. To minimize the risk of contamination, soil samples were collected while wearing protective gloves, sterilized gowns, and masks (Fig. S1). Owing to the fact that our sampling was conducted on soils left exposed at open-air sites, surface soils within a 1-m radius of the site also were sampled as negative controls. When the results on the surface soils showed negativity, the data from the geological strata were regarded as authentic. The archaeological information on the soil samples (n ¼ 143) is summarized in Table 1. In Gyeongju City, the Silla capital, we collected samples from two different archaeological sites: GNM_A and GNM_B. The total number of Gyeongju samples was 67. At the GNM_A site, the samples (n ¼ 58) represented four different subareas (A to D). The sampled strata were date-estimated to the 7th to 9th century (the Unified Silla period in Korean history). Three samples (B to D) were collected from the strata that had been in side gutters on the streets of Gyeongju city when it was the Silla capital. Sewage in those streets almost certainly would have been drained through them (Fig. S2). At the GNM_B site, the nine sediment samples collected had been in a storage place dating to the 7th to 9th century CE (Fig. S3).
Table 1 The archaeological information of the Silla samples used in this study. District
Sites/strata in the district
GNM_A (Gyeongju)
A B C D Seokjo Yugu 1
GNM_B (Gyeongju) Bonghwang-Dong (Gimhae)
Hwawang Sansung (Changnyeong)
Sample number 24 5 17 12 9
Estimated period
Archaeological information
Silla 7the9th Century CE
Not determined for its usage in Silla period Side Gutter in the capital of Silla kingdom
A storing place
Joseon Goryeo Flood Precipitates A
6 3 6
Silla 7the9th Century CE Joseon Goryeo ND
Shell-midden Flood Precipitates B I
6 6
Silla Silla
12
Silla Before unification of Korean peninsular in 676 CE
II
14
Silla 6the9th Century CE
III
10
Silla Before unification of Korean peninsular in 676 CE Joseon Silla
IV V Total
5 8 143
The geological layer of Joseon period The geological layer of Goryeo period The geological layer is situated between those of Goryeo and Silla periods. It could be made by flood overflow. The exact date could not be determined by archaeologists because any cultural remains for date estimation were not found in the layer Shell-midden layer of Silla period The geological layer made by flood overflow. Possibly made during Silla period Sample from Grid 4. The geological strata formed at the interim period of Reservoirs #2 and #3. Corresponded to Ancient Silla kingdom stratum before unification of Korean peninsula (676 CE) Sample from precipitates in Reservoir#2 (inside). The geological strata were formed during the reservoir sill in use. Reservior#2 looks to have been mainly used after unification of Korean peninsula Sample from Reservoir#3 (base). The reservoir layer was formed before unification of Korean peninsula Geological strata formed during Joseon period Geological strata formed during Sillla period
Please cite this article in press as: Kim, M.J., et al., Paleoparasitological study on the soil sediment samples from archaeological sites of ancient Silla Kingdom in Korean peninsula, Quaternary International (2015), http://dx.doi.org/10.1016/j.quaint.2015.02.007
M.J. Kim et al. / Quaternary International xxx (2015) 1e7
Fig. 1. The map of South Korea showing sampling sites in this study. Red-shaded area is Gyeongsang provinces where we performed the sampling for the current study. A, Gyeongju; B, Changnyeong; C, Gimhae. We obtained parasite-egg positive results from the various archaeological sites of the ancient Silla period. The current results form a striking contrast to our previous paleoparasitological study on the samples from southwestern part of South Korea (blue-shaded area). The study in 2011, though we examined 593 samples from 17 sites, showed very few positive findings (Kim et al., 2013). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
At the Bongwhang-Dong site, we conducted sampling from five different geological strata (Joseon, Goryeo, Flood_Precipitates_A, Shell-midden, and Flood_Precipitates_B). The total number of samples was 27. Of these, the Shell-midden (n ¼ 6) and
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Flood_Precipitates_B samples (n ¼ 6) corresponded to the Silla period. The Flood_Precipitates_A strata age could not be estimated, due to historical disturbances by floods. For comparative purposes, we also sampled from Joseon (n ¼ 6) and Goryeo (n ¼ 3) strata (Fig. S4). Additionally, we conducted sampling at an ancient fortress built on the top of Hwawang Mountain. The total number of samples collected there was 49. Sample sets I, II and III represented 6th to 9th century ruins of a reservoir that had been constructed and maintained by the Silla people. Samples I and III were deemed likely to have been formed before the unification of the Korean peninsula (676 CE), while sample II was estimated to have been precipitated post-unification (the Unified Silla period: 676e935 CE). Sample V, another Silla sample, was collected from a site very close to the south wall of the fortress. We also obtained Joseon samples (sample IV, n ¼ 5) for comparison (Fig. S5). The detailed sample information is summarized in Table 1. After transporting the samples to our lab, we prepared them for light-microscopic examination (Callen and Cameron, 1960; Criscione et al., 2007). Briefly, the samples were re-hydrated in 0.5% trisodium phosphate solution and filtered through multiplelayer gauze. After one additional precipitation, the upper turbid layer was discarded, and the precipitates were dissolved in 10% neutral-buffered formalin. The precipitate solutions were then dropped onto slides and examined under light microscopy (Olympus, Japan). A total of 200 ml of each sample (10 ml 20 times each or 20 ml 10 times each) was used. The sizes of the eggs were measured, and the numbers of parasite eggs per gram (EPG) was also estimated. 3. Results There were no parasite eggs discovered in the negative-control samples from the surface soils, ruling out any possibility of false
Fig. 2. Results of paleoparasitological examination on Bongwhang-Dong sample. Top figure is geological strata where we sampled. Dots indicate each sampling point. Yellow and red dots indicate the points showing negative and positive results, respectively. The eggs (bottom) are those identified in the samples. Scale bars ¼ 20 mm. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Please cite this article in press as: Kim, M.J., et al., Paleoparasitological study on the soil sediment samples from archaeological sites of ancient Silla Kingdom in Korean peninsula, Quaternary International (2015), http://dx.doi.org/10.1016/j.quaint.2015.02.007
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positivity caused by accidental introduction of surface soils into the examined geological strata. In the cases of the samples from GNM_A and _B (Gyeongju) representing the Silla period, we could not identify any ancient parasite eggs, even though they had been collected from highly populated areas of the Silla capital. However, in geological-strata samples from Bongwhang-Dong and Hwawang Sansung, we observed many parasite eggs. At the Bongwhang-Dong site, ancient parasite eggs were found in the samples taken from the Goryeo, Flood_Precipitates_A, Shellmidden and Flood_Precipitates_B layers, whereas none was found in the same site's Joseon layer (Fig. 2). The Shell-midden and Food_Precipitates_B layers showed particularly heavy contamination during the Silla period. With regard to the samples from the Hwawang Sansung site, we found that most of the Silla samples (I, II and III) from the fortress reservoir exhibited ancient parasite eggs. On the other hand, we could not find any eggs in either the Joseon (IV) or Silla sample (V) taken from an area very close to the south wall of the same Silla fortress (Fig. 3). In the samples from the Bongwhang-Dong and Hwawang Sansung sites, numerous Ascaris and Trichuris sp. eggs were identified. Interestingly, in those Bongwhang-Dong samples, dicrocoelid eggs also were discovered (Fig. 2); and in the Hwawang Sansung samples, Taenia sp. eggs, a species very rarely detected in South Korea, were found (Fig. 3). The characteristics of Ascaris, Trichuris, Taenia spp. and dicrocoelid eggs observed in this study are well matched with previous reports on the species (Garcia, 2007; Searcey et al., 2013). The egg sizes and EPGs estimated in this study are summarized in Table 2.
from the pre-20th century strata in the Joseon Dynasty's capital area (a part of what is now modern Seoul Metropolitan city). Our discovery of many eggs prompted a suspicion that pre-modern major cities in East Asia might have been seriously contaminated by parasite eggs that almost certainly have originated from the inhabitants' feces (Shin et al., 2011b, 2013b). The remarkable achievements of paleoparasitological studies on ancient samples notwithstanding, it is still disappointing that most discoveries have been at archaeological sites of relatively recent origin. More in-depth studies on samples representing pre-Joseon periods are therefore needed for a comprehensive and complete understanding of Korea's parasitological history. The results of our current investigation of strata and samples from the prosperous firstmillennium era of the Silla Kingdom are therefore very encouraging. In this study, we found many ancient parasite eggs in the samples from the Silla strata of the Hwawang Sansung (a mountaintop fortress reservoir) and Bongwhang-Dong (shell-midden) sites. The Bongwhang-Dong site was informative because we discovered ancient parasite eggs in shell-midden samples from the Silla period. In Japan, Matsui et al. (2003) observed ancient parasite eggs in samples from a shell-midden site that had been a garbage dump dating to the prehistoric Jomon period. The authors attributed the presence of the eggs there to the shell-midden site's possible role as a kind of toilet in pre-historic Jomon society (Matsui et al., 2003). Our detection of ancient parasite eggs in shell-midden samples from the Silla period thus indicates that the East Asian people still used the midden as a substitute or complementary toilet, or at least, that they might have dumped night soils there, even after the end of the pre-historic age.
Table 2 Positive results of paleoparasitological examinations in this study. Sites
Area/strata in the site
Estimated period
Parasite eggs identified
Average dimensions
Bonghwang-Dong (Gimhae)
Goryeo
Goryeo 935e1392 ND
Trichuris trichiura
51.2 26.6
1.6
Ascaris lumbricoides Trichuris trichiura
55.0 50.0 50.2 22.5
0.8 3.3
Ascaris lumbricoides Trichuris trichiura Ascaris lumbricoides Trichuris trichiura Dicrocoelid Ascaris lumbricoides Trichuris trichiura Taenia spp. Ascaris lumbricoides Trichuris trichiura Trichuris trichiura Ascaris lumbricoides Taenia spp.
61.5 46.0 47.4 25.6 60.0 47.5 50.3 26.9 41.7 26.0 62.4 46.3 49.0 25.2 39.8 38.6 58.3 50.0 47.5 24.5 50.0 25.0 60.0 43.9 35.0 31.0
5.0 0.8 3.3 11.6 2.5 5.0 5.6 0.4 3.2 5.4 2.5 7.0 0.5
Flood Precipitates A Shell-midden
Hwawang Sansung (Changnyeong)
Silla
Flood Precipitates B I
Silla
II
Silla 6the9th Century CE Silla Before unification of Korean peninsular in 676 CE
III
Silla Before unification of Korean peninsular in 676 CE
4. Discussion In general, heavy parasite-egg contamination of soil is known to correlate well with higher parasitic infection prevalence among inhabitants of the given area (Schulz and Kroeger, 1992; Uga et al., 1995, 1997; Chongsuvivatwong et al., 1999). This is no less true of ancient samples from hundred-to-thousand year-old archaeological sites. It is highly probable that detection of parasite eggs in soil samples from ancient or medieval towns or cities will correlate very closely with the parasitic infection patterns among the relevant historical peoples (Monckton, 1995; Matsui et al., 2003; Fernandes et al., 2005; da Rocha et al., 2006; Mitchell et al., 2008). We have experienced such situations in our studies on ancient samples obtained from a variety of archaeological sites in South Korea. One case involved the examination of soil-sediment samples
EPG
Despite the contributions of the studies just noted, information on shell-midden sites in East Asian history, from the perspective of paleopathology at least, remains largely incomplete. We still do not have any idea of when the use of shell-midden sites finally disappeared in East Asian countries. Did the transition coincide with the adoption of more advanced forms of toilets? Only future studies could answer such a question. Next, we discuss the outcomes of the examinations of the Hwawang-Sansung fortress samples. In ancient Korea, many fortresses were built, in every corner of the country and especially on mountaintop areas. These were refuges in times of war, though recent historical studies have revealed that they were more than that as well. Mountain fortresses were established political centers that were significantly populated even in peacetime (Seo, 2010, 2012). Considering the semi-permanent statuses of such
Please cite this article in press as: Kim, M.J., et al., Paleoparasitological study on the soil sediment samples from archaeological sites of ancient Silla Kingdom in Korean peninsula, Quaternary International (2015), http://dx.doi.org/10.1016/j.quaint.2015.02.007
M.J. Kim et al. / Quaternary International xxx (2015) 1e7
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Fig. 3. Results of paleoparasitological examination on Hwawang Sansung samples. The figures for archeological sites represent each area (I to V) where we sampled. Yellow and red dots indicate the sampling points showing negative and positive results, respectively. SW, south wall of the fortress. The bottom figures of parasite eggs are those found in the ancient samples I, II and III. Scale bars ¼ 10 mm. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
fortresses, their inner areas must have been to some extents contaminated by human wastes, and therefore, at least relatively unsanitary. The evidence gathered in the current study supported this speculation. For example, in the fortress reservoir samples dating to the Silla period, we detected ancient parasite eggs. As to the question of why parasite eggs were plentifully identified in the ancient reservoir samples, previous research on ancient moats around permanent settlements can be very informative. For instance, upon our detection of ancient parasite eggs in the samples from a moat constructed around the Silla King's palace in Gyeongju city, we conjectured that the phenomenon might have been caused by a continual feces-contaminated inflow, from the King's palace into the surrounding moat area (Shin et al., 2009c). Inflow-contamination of an ancient moat encircling a permanent settlement also has been reported in Japan. Matsui et al. (2003) found evidence that wastewater or excrement from a settlement dating to the Yayoi period (450 BCEe300 CE) had been continuously discharged into the surrounding moat. Although the stagnant water in the moat might have made for serious sanitary
problems, this system seemingly was indispensable to them in any case, as human waste had to be removed from the settlement somehow. Like the discharging inflows to ancient moats around the settlements (Matsui et al., 2003; Shin et al., 2009c), creeks or streams in the fortress must have flown into the reservoir, which had been constructed at a lower altitude. Human wastes scattered in the fortress and riding on the stream flows could also have been drained into the reservoir. The ancient eggs finally detected in the reservoir samples could well have originated in the feces. Based on our study, we speculated that parasite re-infection must have been highly prevalent among the fortress-dwellers; and the eggs laid by the adult parasites must have been defecated into the soil of the region, contaminating the fortress area continuously during the ancient Silla period. Of the ancient parasite eggs discovered in various Silla samples of this study, dicrocoelid and Taenia eggs have been reported only very rarely in previous works on ancient samples in South Korea (Lee et al., 2011). Dicrocoelium sp. has attracted
Please cite this article in press as: Kim, M.J., et al., Paleoparasitological study on the soil sediment samples from archaeological sites of ancient Silla Kingdom in Korean peninsula, Quaternary International (2015), http://dx.doi.org/10.1016/j.quaint.2015.02.007
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M.J. Kim et al. / Quaternary International xxx (2015) 1e7
paleoparasitologists' interest for its importance to understanding of historical human migrations that have occurred between continents. The detection of dicrocoelid eggs in Western Europe was reported in samples dating to as early as 550,000 years BCE to the 16th century CE, while New World samples began to exhibit the same species at around the 17th century CE, after European colonization (Le Bailly and Bouchet, 2010). However, the number of relevant reports is still not sufficient to constitute a global overview of ancient dicrocoeliosis history. The reports on the presence of ancient Dicrocoelium eggs in European, North American and African archaeological samples have been sporadic (Dittmar and Steyn, 2004). In Asia, no reports on ancient Dicrocoelium eggs have been available until now. In this regard, our detection of Dicrocoelium eggs in ancient Silla samples, the firstever such report for Asia, is very meaningful. At least, this clearly shows that, like any other Old World peoples, the dicrocoelid also infected Asians even before the Columbian discovery of America. Another noteworthy finding in the present study was the detection of Taenia eggs in the Hwawang Sansung fortress samples. According to Matsui et al. (2003), Taenia sp. has been very rarely detected in ancient Japanese samples. An exception was the sample from the cesspits at the Koro-Kan site, the 8th century guesthouse in Japan. About the presence of ancient Taenia sp. eggs in the cesspit samples, Matsui et al. (2003) speculated that the eggs might not have originated from Japanese, but rather, that they could have been deposited by diplomatic delegates from foreign countries. The authors' hypothesis was clearly based upon the speculation that the people in neighboring countries, namely China or Korea, might have been more vulnerable to taeniasis by enjoying pork and beef much freely than could the ancient Japanese. In Japan during the ancient period, meat-eating was strictly prohibited for religious reasons (Matsui et al., 2003). However, Taenia sp. eggs from ancient East Asian samples have only rarely been reported. Among the multiple studies and many samples of the past several years, only one sample, taken from a Joseon tomb, contained Taenia sp. eggs (Lee et al., 2011). Moreover, given the lack of any reports on the presence of ancient Taenia eggs in South Korean samples similar in age to those in the Koro-Kan case, Matsui et al. (2003)'s speculation that the Taenia sp. eggs detected in ancient Japanese samples had actually originated from visiting Korean or Chinese delegates was a subject for further studies. In this regard, the detection of Taenia sp. eggs in the Silla samples is very interesting. According to the current results, it could not be completely ruled out that Taenia sp. infection was prevalent among Korean populations even in ancient times. However, in any case, the hypothesis requires further corroboration in forthcoming research. For instance, the difference in prevalence between ancient Korean and Japanese peoples must also be clarified by future paleoparasitological studies. 5. Conclusions Given the late start of paleoparasitological studies in Korea, fundamental data necessary for any reasonably complete understanding of parasitic infection patterns are still insufficient. Our present series of paleoparasitological examinations of ancient Korean archaeological samples hopefully will go some way toward supplementing that data. In our study, we analyzed soil-sediment samples obtained from sites dating to the Silla Kingdom (57 BCEe935 CE). Specifically, Ascaris lumbricoides, Trichuris trichiura, Taenia spp. and dicrocoelid eggs were observed in a shellmidden and at a mountaintop fortress. We were able to derive invaluable data on parasitic infection patterns of ancient Koreans, information that previously was difficult to obtain.
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Please cite this article in press as: Kim, M.J., et al., Paleoparasitological study on the soil sediment samples from archaeological sites of ancient Silla Kingdom in Korean peninsula, Quaternary International (2015), http://dx.doi.org/10.1016/j.quaint.2015.02.007