Molecular insights into the history of plague
Michel Drancourt, and Didier Raoult
Unité des rickettsies, CNRS UPRES-A 6020, faculté de médecine, université de la Méditerranée, 27 boulevard Jean Moulin, 13385 Marseille cedex 5, France
Microbes and Infection
Volume 4, Issue 1 , January 2002, Pages 105-109
Article Outline
1. Historical descriptions of plague
4. Retrospective diagnosis of plague
The history of plague is often as confusing as the recorded history of mankind itself. There are numerous references to plagues that may have been due to Yersinia pestis in ancient texts, including the Old Testament. The first recorded outbreak of an epidemic consistent with plague, however, was in
After an absence of plague for 600 years in
2. The microbiology of plague
During the Hong Kong epidemic in June 1894, Alexandre Yersin 10 and Shibasaburo Kitasato independently announced within a few days of one another the isolation of the plague organism, which was named Yersinia pestis in 1944 11 and 12. Yersin gave us a clear description of bubonic plague in
3. The limits of historical descriptions of plague and controversial issues regarding the history of the disease
Historical descriptions of plague are found in paintings and in old texts which provide epidemiological and clinical descriptions of the disease. The interpretation of historical texts is often limited by the absence of the original text itself, problems with the translation of ancient words into contemporary language and a lack of precise medical terms in old languages. Also, until the second part of 19th century, confusion existed between diseases presenting with similar signs and symptoms, for example typhus and typhoid fever 30. In historical documents, a description of an epidemic associated with high mortality rates that occur within a few days and the presence of buboes probably has the highest predictive value for plague. Although encountered in other contemporary infectious diseases, buboes––large, painful, engorged lymph nodes––are a useful sign to distinguish plague. None of the above three characteristics alone, however, is sufficient for a diagnosis of plague, and this has caused controversies regarding the etiology and the epidemiology of the disease 31. In particular, for suspected plague epidemics that occurred before 1894, only degrees of probability can be offered as to whether these were in fact caused by Y. pestis. Descriptions of the plague of Athens for example 32 are not consistent with any disease we know and bubonic plague, typhus, smallpox 33, staphylococcal toxic shock syndrome-complicated influenza 34, Ebola fever 32, melioidosis 35 and other diseases 36 have been regarded as the diseases probably involved. Also, as the high levels of mortality and transmissibility associated with the Black Death were not observed during the third plague pandemic, controversies arose regarding the etiology of the Black Death, and some historians believed it was not plague. The alternatives have included anthrax, typhus, tuberculosis and hemorrhagic fever 8; 37 and 38. As we show below, modern molecular methods have enabled us to demonstrate clearly that the second pandemic was in fact due to Y. pestis 39 and 40 Controversies have also arisen regarding the reservoirs in epidemics of plague. Yersin established that the black rat, Rattus rattus was the reservoir of Y. pestis during the third pandemic 10. Observation that some plague epidemics lacked an obvious murine reservoir, however, led to the discovery of telluric resistance Y. pestis 14. Changes in the numbers and geographical distribution of R. rattus populations have been found to correlate with plague epidemics during the first and second pandemics, supporting the role of this species as a reservoir for Y. pestis during these periods. Several workers described the presence of R. rattus on the southern coasts of the Mediterranean sea in the ancient world and the rat was confirmed to be present in Corsica, the Baleares Islands and Pompei in the 4th–2nd century BC 41. R. rattus developed a close association with humans during the Roman expansion and the early Middle Ages but remained localized in small territories in urban areas until the 11th century. Archeological data from Antiquity indicate that black rats migrated from Mediterranean harbors along the routes and their geographical distribution matched that of the Justinian plague 42. During the second millenium, economic prosperity resulted in a rapid expansion of trade between European and Eastern countries, and between southern and northern European countries. The R. rattus population grew very rapidly and this species has now been found in almost all the archeological sites in
4. Retrospective diagnosis of plague
Molecular biology tools enable the detection of microbial genome fragments in ancient human remains and thus the possibility of making retrospective diagnoses of ancient diseases. By polymerase chain reaction (PCR) sections of microbial DNA may be amplified enzymatically and then sequenced to assess the percentage of similarity between parts of the genomes of ancient microorganisms and those of their modern-day counterparts deposited in electronic databases. DNA is known to persist for long periods after the death of an organism 47. There are, however, chemical modifications and fragmentation of the ancient DNA 48 that may occur and limit the application of the above techniques. Also, uncharacterized inhibitors of the PCR may be present in ancient DNA samples 49. Molecular tools were first applied to detect mycobacterial DNA in ancient human corpses that had macroscopic signs of either tuberculosis 50 or leprosy 51. Mummified tissues 51 and 52 or, more frequently, remnants of bones have been used to detect microorganisms that might have infected the person 48; 53 and 54. While mummified tissues are rare, bones can be found more readily but require time-consuming decalcification processing before DNA extraction can be carried out. During these procedures the samples may be contaminated with microorganisms present in the environment, a particular problem when trying to detect Y. pestis, which may be present in soil. Also, bone samples are best used to detect microorganisms that cause lesions within the bones themselves. For example in syphilis or tuberculosis, where the causative bacteria may multiply in the bones and therefore multiple DNA copies are available for amplification. Where septicemia is a feature of the disease, such as plague, bone may not be a good sample as there may only be very few bacteria present.
In humans with plague, death usually occurs during the septicemic phase of the disease and virtually all well-vascularized tissues are contaminated by Y. pestis, including the dental pulp. We therefore postulated that dental pulp, by virtue of its good vascularization, durability and natural sterility, would be a suitable sample on which to attempt the demonstration of Y. pestis by molecular biology techniques. We have tested our hypothesis on other organisms causing septicemia and have detected Coxiella burnetii DNA in dental pulp extracted from experimentally infected mice 55 and Rickettsia rickettsii DNA in the dental pulp of experimentally infected guinea-pigs (unpublished data). To determine if this was also possible for Y. pestis, we extracted DNA from the dental pulp of teeth extracted from skeletal remains of people suspected to have died of plague 39. Molecular targets that we attempted to amplify included the plasmid-encoded pla gene, encoding a virulence factor of Y. pestis, and the rpoB gene, encoding the beta-subunit of the bacterial RNA polymerase, which is a molecular target used for the identification of enteric bacteria 56. In our first set of experiments, dental pulp was extracted from the teeth of three skeletal remains of people suspected of having died of plague in Marseilles in 1722 and from two corpses buried in 1590 in Lambesc, a southern French village where there had also been a plague outbreak 39. As negative controls, dental pulp was extracted from skeletal remains in medieval tombs in
5. Conclusions
Combining the use of dental pulp as a source of DNA and our `suicide PCR' protocol may facilitate the detection of the DNA of ancient pathogens responsible for septicemic diseases in the past. Cremation, as practiced by Greeks during the
J.N. Biraben In: Les hommes et la peste en
J.N. Biraben and J. Le Goff , The plague in the early middle ages. In: R Forster and O Ranum, Editors, Biology of Man in History, The Johns Hopkins University Press, Baltimore, MD (1975), pp. 48–80.
R. Pollitzer Plague, World Health Organization,
V.J. Derbes and De , Mussis and the Great Plague of 1348. A forgotten episode of bacterial warfare. JAMA 196 (1966), pp. 179–182.
De Mussis G., Historia de morbo. Mortalitate quae fuit anno Dei MCCCXLVIII, original manuscript in codex 59, library of the University of Wroclaw, Poland.
P. Ziegler The Black Death, Alan Sutton Publishing Inc, Wolfeboro Falls, NH (1991).
J. Enselme , Gloses sur le passage dans la ville d'Avignon de la grande mortalité de 1348. Rev. Lyon M 17 (1969), pp. 697–710. Abstract-MEDLINE | $Order Document
S. Scott, C.J. Duncan and S.R. Duncan , The plague in
M. Signoli, J. da Silva, E. Georgeon, G. Léonetti and O. Dutour , Verification of death during the Great Plague of Marseilles: anthropological data from the excavation of mass grave of `l'Observance'. C.R. Acad. Sci. Paris 322 (1996), pp. 333–339. Abstract-GEOBASE | $Order Document
A. Yersin , La peste bubonique à
L. Gross , How the plague bacillus and its transmission through fleas were discovered: reminiscences from my years at the Pasteur Institute in
J.J. van Loghem , The classification of the plague bacillus, Antonie van Leeuwenhoeck. J. Serol. Microbiol. 10 (1944), pp. 15–16.
P.L. Simond , La propagation de la peste. Ann. Inst. Pasteur (
H. Mollaret , Conservation expérimentale de la peste dans le sol. Bull. Soc. Pathol. Exot. 6 (1963), pp. 1169–1183.
Y. Karimi , Conservation naturelle de la peste dans le sol. Bull. Soc. Pathol. Exotique 6 (1963), pp. 1183–1186.
M. Baltazard , Evolution de la recherche sur l'épidémiologie de la peste. Med. Mal. Infect. 1 (1971), pp. 203–218.
Human plague in 1994. Wkly Epidemiol. Rec. 71 (1996), pp. 165–168.
A.K. Akiev , Epidemiology and incidence of plague in the world, 1958–1979. Bull. WHO 60 (1982), pp. 165–169. Abstract-MEDLINE | Abstract-EMBASE | $Order Document
Human plague – United States, 1993–1994. MMWR Morb. Mortal. Wkly Rep. 43 (1994), pp. 242–246.
N.S. Deodhar, V.L. Yemul and K. Banerjee , Plague that never was: a review of the alleged plague outbreaks in 1994.
A. Guiyoule, B. Rasoamanana, C. Buchrieser, P. Michel, S. Chanteau and E. Carniel , Recent emergence of new variants of Yersinia pestis in Madagascar. J. Clin. Microbiol. 35 (1997), pp. 2826–2833. Abstract-MEDLINE | Abstract-EMBASE | $Order Document
M. Galimand, A. Guiyoule, G. Gerbaud, B. Rasoamanana, S. Chanteau, E. Carniel and P. Couvalin , Multidrug resistance in Yersinia pestis mediated by a transferable plasmid. N. Engl. J. M 337 (1997), pp. 677–680. Abstract-MEDLINE | Abstract-EMBASE | $Order Document
D.T. Dennis and J.M. Hughes , Multidrug resistance in plague. N. Engl. J. M 337 (1997), pp. 702–704. Abstract-MEDLINE | Abstract-EMBASE | $Order Document | Full Text via CrossRef
A. Ibrahim, B.M. Goebel, W. Liesack, M. Griffiths and E. Stackebrandt , The phylogeny of the genus Yersinia based on 16S rDNA sequences. FEMS Microbiol. Lett. 114 (1993), pp. 173–178. Abstract-Elsevier BIOBASE | $Order Document
H. Bercovier, H.H. Mollaret, J.M. Alonso, J. Brault, G.R. Fanning, A.G. Steigerwalt and D.J. Brenner , Intra- and interspecies relatedness of Yersinia pestis by DNA hybridization and its relationship to Y. pseudotuberculosis. Curr. Microbiol. 4 (1980), pp. 225–229. Abstract-EMBASE | $Order Document
M. Achtman, K. Zurth, G. Morelli, G. Torrea, A. Guiyoule and E. Carniel , Yersinia pestis, the cause of plague, is a recently emerged clone of Yersinia pseudotuberculosis. Proc. Natl. Acad. Sci. 96 (1999), pp. 14043–14048. Abstract-MEDLINE | Abstract-EMBASE | Abstract-Elsevier BIOBASE | $Order Document | Full Text via CrossRef
R. Davignat , Variétés de l'espèce Pasteurella pestis. Nouvelle hypothèse. Bull. OMS 4 (1951), pp. 247–263.
A. Guiyoule, F. Grimont, I. Iteman, P.A.D. Grimont, M. Lefèvre and E. Carniel , Plague pandemics investigated by ribotyping of Yersinia pestis strains. J. Clin. Microbiol. 32 (1994), pp. 634–641. Abstract-MEDLINE | Abstract-Elsevier BIOBASE | Abstract-EMBASE | $Order Document
L.T. Wu, J.W.H. Chun, R. Pollitzer and C.Y. Wu Plague: a Manual for Medical and Public Health Workers,
W. Jenner On the Identity or Non-identity of Typhoid and Typhus Fevers, C & J Adlard,
P. Slack , The Black Death past and present. 2. Some historical problems. Trans. Roy. Soc. Trop. Hyg. 83 (1989), pp. 461–463. Abstract-MEDLINE | Abstract-EMBASE | $Order Document
P.E. Olson, C.S. Hames, A.S. Benenson and E.N. Genovese , The Thucydides syndrome : Ebola déjà vu ? (or Ebola reemergent ?). Emerg. Infect. Dis. 2 (1996), pp. 155–156. Abstract-MEDLINE | $Order Document
F.P. Retief and L. Cilliers , The epidemic of
A.D. Langmuir, T.D. Worthen, J. Solomon, C.G. Ray and E. Peterson , The Thucydides syndrome : a new hypothesis for the cause of the plague of
E.M. David , "The good and the bad dying indiscriminately": the Athenian plague reconsidered. Pharos of Alpha Omega Alpha Spring 63 (2000), pp. 4–7. Abstract-MEDLINE | $Order Document
A.J. Holladay , The Thucydides syndrome: another view. N. Engl. J. M 315 (1986), pp. 1170–1173. Abstract-MEDLINE | $Order Document
G. Twigg The Black Death. A biological reappraisal, Bratsford,
E. Weiss , Rickettsias. In: J Lederberg, Editor, Encyclopedia of Microbiology, Academic,
M. Drancourt, G. Aboudharam, M. Signoli, O. Dutour and D. Raoult , Detection of 400-year-old Yersinia pestis DNA in human dental pulp. An approach to the diagnosis of ancient septicemia. Proc. Natl. Acad. Sci. USA 95 (1998), pp. 12637–12640. Abstract-MEDLINE | $Order Document | Full Text via CrossRef
D. Raoult, G. Aboudharam, E. Crubezy, G. Larrouy, B. Ludes and M. Drancourt , Suicide amplification of the medieval Black Death bacillus. Proc. Natl. Acad. Sci. USA 97 (2000), pp. 12800–12803. Abstract-MEDLINE | Abstract-EMBASE | Abstract-Elsevier BIOBASE | $Order Document | Full Text via CrossRef
F. Audoin-Rouzeau , Le rat noir (Rattus rattus) et la peste dans l'Occident antique et médiéval. Bull. Soc. Pathol. Exot. 92 (1999), pp. 422–426. Abstract-MEDLINE | $Order Document
J.N. Biraben and J. Le Goff , La peste dans le haut Moyen Age. In: A Collin, Editor, Annales E.S.C., vol. 24,
G. Blanc and M. Baltazard , Documents sur la peste. Arch. Inst. Pasteur Maroc 3 (1945), pp. 349–354.
E.A. Eckert , Seasonality of plague in early modern
P. Slack The Impact of Plague in Tudor and Stuart England, Routledge,
J.C. Beaucournu , A propos du vecteur de la peste en
S. Pääbo , Molecular cloning of ancient Egyptian mummy DNA. Nature 314 (1985), pp. 644–645. Abstract-EMBASE | Abstract-MEDLINE | $Order Document
S. Pääbo , Ancient DNA: extraction, characterization, molecular cloning, and enzymatic amplification. Proc. Natl. Acad. Sci. USA 86 (1989), pp. 1939–1943. Abstract-MEDLINE | Abstract-EMBASE | $Order Document
C. Hänni, T. Brousseau, V. Laudet and D. Stehelin , Isopropanol precipitation removes PCR inhibitors from ancient bone extracts. Nucl. Acids. Res. 23 (1995), pp. 881–882. Abstract-MEDLINE | Abstract-Elsevier BIOBASE | Abstract-EMBASE | $Order Document
G.M. Taylor, M. Goyal, A.J. Legge, R.J. Shaw and D. Young , Genotypic analysis of Mycobacterium tuberculosis from medieval human remains. Microbiology 145 (1999), pp. 899–904. Abstract-EMBASE | Abstract-MEDLINE | $Order Document
A. Rafi, M. Spigelman, J. Standford, E. Lemma and H. Donoghue , Zias J., Mycobacterium leprae DNA from ancient bone detected by PCR. Lancet 343 (1994), pp. 1360–1361. Abstract | Full Text + Links | PDF (331 K)
A. Nerlich, C.J. Haas, A. Zink, U. Szeimies and H.G. Hagedorn , Molecular evidence for tuberculosis in ancient Egyptian mummy. Lancet 350 (1997), pp. 1404–1404. Abstract-EMBASE | Abstract-MEDLINE | $Order Document
M. Spigelman and E. Lemma , The use of the polymerase chain reaction to detect Mycobacterium tuberculosis DNA in ancient skeletons. Int. J. Osteoarcheol. 3 (1993), pp. 137–143.
G.M. Taylor, M. Crossey, J.A. Saldanha and T. Waldron , Detection of Mycobacterium tuberculosis bacterial DNA in medieval skeletal remains using polymerase chain reaction. J. Archeol. Sci. 23 (1996), pp. 789–798.
G. Aboudharam, B. Lascola, D. Raoult and M. Drancourt , Detection of Coxiella burnetii DNA in dental pulp during experimental bacteremia. Microb. Pathog. 28 (2000), pp. 249–254. Abstract | Abstract + References | PDF (290 K)