Molecular Confirmation of Yersinia pestis in 6th century Bavaria

Erasing any lingering doubts about the agent of the Plague of Justinian, a group of German biological anthropologists have shown conclusively that Yersinia pestis caused an epidemic in a 6th century Bavarian cemetery at Aschheim. Harbeck et al (2013) provide a convincing refutation of previous theories about the etiologic agent of the Plague of Justinian.   Returning to the same cemetery where plague was previously reported, two independent labs using the most modern standards to prevent contamination confirmed Yersinia pestis from multiple burials within the cemetery making this the best characterized Early Medieval plague cemetery.

The cemetery, called Aschheim, is in Bavaria outside of Munich. It contains the remains of 438 people with an unusually high number of multiple graves but no disordered mass graves. The 19 multiple burials contained two to five individuals arranged in lines. The cemetery was dated archaeologically to 500-700 AD with remains being carbon dated ranging from 530 to 680, all consistent with the 541 pandemic and its aftermath. Harbeck et al (2013) tested 19 individuals from 12 multiple graves. From these, there were eight positive samples, but only one produced enough aDNA to do some SNP genotyping. Added to the previous paper, this makes 11 positive individuals from this cemetery. Given the tenuous survival of aDNA, 11 positive individuals out of 21 tested in the two combined papers is a very good success rate. This is a cemetery that the F1 antigen test would be interesting since it could be used on the entire cemetery without great cost or labor. More sensitive than aDNA, the antigen test could tell us the percentage of plague deaths in the cemetery.

Individual A120 was screened with several SNPs that mapped it to an early region of the phylogenetic tree in the 0.ANT section. This makes the Plague of Justinian isolate ancestral to the Black Death isolates (yellow boxes below) from East Smithfield. This section whose only point of diversity is 0.ANT1 at node 4. Date predictions for the nodes of diversity in the tree fits with the Plague of Justinian falling in this region.  Modern isolates that  form this region of the phylogenetic tree all come from central Asia (around Tibet), suggesting that like the Black Death, the Plague of Justinian also originated in Asia. Overall, everything fits in well with expectations for the first pandemic.

(Harbeck et al, 2013. Fig. 1)

(Harbeck et al, 2013. Fig. 1)

Reference:

Harbeck M, Seifert L, Hänsch S, Wagner DM, Birdsell D, et al. (2013) Yersinia pestis DNA from Skeletal Remains from the 6th Century AD Reveals Insights into Justinianic Plague. PLoS Pathog 9(5): e1003349. doi:10.1371/journal.ppat.1003349

Wiechmann I, & Grupe G (2005). Detection of Yersinia pestis DNA in two early medieval skeletal finds from Aschheim (Upper Bavaria, 6th century A.D.). American journal of physical anthropology, 126 (1), 48-55 PMID: 15386257

The Dancing Plague of 1518


John Waller. The Dancing Plague: The Strange, True Story of an Extraordinary Illness. Sourcebooks, 2009 (paperback). Previously published as A Time to Dance, A Time to Die: The Extraordinary Story of the Dancing Plague of 1518 (Icon books, hardback, 2008).

Topic: Dancing Mania, choreomania

Time and Place: Strasbourg, Holy Roman Empire, 1518

Audience: General

Discussion:

This was a good book to wrap up 2011 with – from the Arab Spring, to summer revolutions and the fall’s occupy movement – conditions are approaching those of 1518. Among the peasants of Strasbourg, life in the early 16th century had become miserable. The church and monasteries left the peasants spiritually desolate and literally starving while their tithes of grain were sold at prices they couldn’t afford. Poor harvests and high inflation made famine a real possibility. There were several peasant revolts leading up to 1518 that were brutally put down with public executions and massacres. Ancient ties between the nobility, church and common people had broken down such that secular and ecclesiastic office holders no longer seemed to care for the welfare of the common people.

In July 1518 Frau Toffea was outdoors in the street of Strasbourg when she began to dance. There was no music or joy in her dancing. The spectacle soon became horrifying as it became apparent that she could not stop. Eventually she collapsed from the heat and exhaustion. When she woke up, she began to dance again in the same frenzy. Soon other people in Strasbourg began their trance-like dancing ignoring the heat and the need for food and water. When they were able to speak, they begged bystanders to make them stop. Before long in the July heat, people began dropping not to get up again – did they dance to an exhausted death or was there a pathological cause?

The people of Strasbourg didn’t see this as a psychological disorder but either physical or spiritual. At first they turned to the newly respected, university trained physicians who strangely proscribed more dancing as a cure. They assigned people to make sure the afflicted kept dancing even when they were capable of stopping. It became very apparent that this was not helping the death rate. Before long physicians and townspeople agreed that this was a spiritual ailment that required a pilgrimage to the local shrine of St Vitas. Fascinatingly the people believed that a saint patronized by epileptics could/would also curse people to uncontrolled movements like the dancing if they were displeased. Throughout this region of Germany and the Netherlands there are shrines to Vitas and other saints to prevent epilepsy and the dancing plague. Late medieval people greatly feared epilepsy and similar disorders because it could be perceived as demonic possession or a curse (by victim as well as bystander). Waller argues that this was something like a case of spiritual post traumatic stress disorder. His argument is too complicated for me to explain here but it is a worthwhile and thought provoking discussion.

Narrative: B+ The narrative was well written and kept me eager to continue reading. This is no small feat without a central cast of characters to follow through the book. He tries to use Frau Troffea as a continuing theme but just doesn’t have enough information to really flesh out her life.  There were places that I wished for more scientific context. I wish he had not saved so much of the science for the last chapter.

Historical Content: A- This is not a time period that I am very familiar so it is hard to assess how well he covered the historical questions. While he discussed actions of the church, he apparently didn’t have sources from within the church. This seems strange as the church usually has better resources than secular courts. The discussion of the religious context was good and mostly from the lay viewpoint.

Scientific Content: B-/C+ He seems to be reaching too far in some of the psychological parallels he tries to draw to the choreomania (dancing plague). Practices that intentionally create trances or mystical dancing are not good matches to the unintentional and unwanted dancing in 1518. The contagious nature of the dancing of 1518 is based at least partially upon the fear of the mania. Waller describes all mystical experiences in terms of pathology, which seems unwarranted. His brief discussion of an outbreak of uncontrollable laughing in Tanzania in 1963 left me wanting to know more (p. 216). A recent Ugandan outbreak of “nodding syndrome” where youths display uncontrollable nodding when they try to eat has been associated with a parasite that causes river blindness. In all three outbreaks, these may be related to epileptic-like behavior. My point here is that we can’t jump to the conclusion that initial cases are all psychological, even if some of the contagious nature does seem psychological (people being effected by watching etc).

I also have to take issue with a reviewers quote the publisher put on the back of the book. A quote from New Scientist stated “It’s a book to make you grateful for the historical increase in human sanity.” Part of the author’s argument is that manifestations of stress and PTSD are culturally dependent and that we express stress differently today. When we consider how many people today take medication for anxiety/stress, depression, PTSD or other psychological conditions it is questionable if there has been a “historical increase in human sanity” – not that manifestations of stress are necessarily measures of sanity anyway.

References and Usability: B The bibliography and notes are integrated together. This makes the bibliography much more difficult to use. The notes are consecutively numbered for the entire book and were a little sparse.  There are 238 notes for 231 pages of text. There were plenty of places I would have liked to have seen that he had a reference for a fact.

Illustrations:  B The maps and illustrations included were okay. It could have used a map of the local Strasbourg area that included the shrine of St Vitas.

Overall, I did enjoy the book and it is a very interesting episode in medical history. Community reaction to the outbreak is as interesting and influential as the disorder itself.  The power of the brain is almost always underestimated. Even if it wasn’t completely psychological, it manifested in ways that were surely under the control of the brain.

A tangent:  one of my favorite quotes from Waller’s book is not on the dancing mania at all. “Syphilis was the flagellum Dei, God’s whip, a stark warning about the sinfulness of adultery and fornication.” If it was the flagellum Dei before the invention of the microscope, imagine how they would have reacted to seeing a spirochete!

Pausing for Peace

Christmas is supposed to be a time of peace and good will toward all men. The Christmas spirit can be found in the most unlikely of places. Perhaps it is in the middle of these places that Christmas is yearned for more than anywhere else. Such was the case on Christmas eve 1914 when both sides agreed to pause for peace and fellowship in the midst of No Man’s Land during the war to end all wars.  Let us commemorate that remarkable event with a you-tube that I hope becomes a classic.

Trench Fever in German Mass Burial

Trench fever seems to be all the rage these days in paleomicrobiology. It seems as though every time Bartonella quintana is added to a panel of pathogens for aDNA screening its found at some level. So far its been found in in a tooth from 4000 before present, in late medieval Venice, 14th century France, and Napoleonic Europe.

Construction at the University of Kassel in Germany discovered a mass grave  revealing “most of the individuals had been males of the age classes juvenis and adultas“. Grumbkow et al (2011) report that local historians, anthropologists and medical examiners concluded that it was a military cohort that died of epidemic disease in the late 18th to early 19th century. They focused on an reputed outbreak of typhoid fever in the winter of 1813/14 that had been linked with regional epidemics started by Napoleon’s troops fleeing from the battle of Leipzig. However, Grumbkow et al (2011) do not present any evidence to directly link their sample of remains to any army that took part in the battle, nor the reasoning behind dating the remains to late 18th to early 19th century for that matter.

To investigate whether these people were victims of a typhoid fever outbreak, Grumbkow et al obtained samples of 18 skeletons for DNA screening. They note that typhoid fever was a common diagnosis for any fever that caused red spots, but could include typhus, parathyroid fever and trench fever. Therefore they screened DNA extracted from 16 femurs and 2 humeri for Salmonella species, Salmonella enterica typhi, Bartonella quintana, Rickettsia prowazekii, and for the bacterial 16S rDNA (ribosomal DNA). As you might have guessed, they only found Bartonella quintana in three bones, that is 16.6.%. This is roughly what Roualt et al (2006) found at Vilnius at 20% for Bartonella quintana.

However, I do have a few concerns with this paper. First, the work is incomplete. They mention in their discussion that they need to screen for more Bartonella quintana genes to confirm their results. This is especially important because the samples they did amplify were all 100% identical to the genebank sequences (making contamination more likely). They list the 16S rDNA primers along with the others but never present any data for these primers. They used diluted “bacterial-positive control DNA” as a control for PCR inhibitors from the soil, which may be what the 16S rDNA primers were used for but it is not explicit. Contamination concerns usually preclude the use of positive controls but they write that “amplification with positive controls and spiked samples always showed the expected results”. The near standard ‘suicide PCR’ protocol for aDNA was not used. Brief communication or not, more information is needed.

ResearchBlogging.org

Grumbkow, P., Zipp, A., Seidenberg, V., Fehren-Schmitz, L., Kempf, V., Groß, U., & Hummel, S. (2011). Brief communication: Evidence of Bartonella quintana infections in skeletons of a historical mass grave in Kassel, Germany American Journal of Physical Anthropology DOI: 10.1002/ajpa.21551  [Epub ahead of print]

A Plague Crypt from Late Medieval Bavaria

St Leonard Catholic Church in Machnung-Pichl, near Ingolstadt Bavaria, Germany held a secret for many years. Renovations to the church back in 1984 found a mass burial site under the sacristy, 75 human skeletons stacked like lasagna in four layers with a little dirt between each layer (Wiechmann, Harbeck, & Grupe, 2010). The design of the site is a little unclear. They say it was not a dug pit and that it can only be dated to 1200 to 1500 CE by the Gothic design of surrounding building structures. From this I conclude that it was some kind of crypt, even if the structure wasn’t originally intended to be a crypt.  I don’t understand why the skeletons couldn’t be carbon dated but it apparently wasn’t done. Without more precise dates, we can’t be sure that all of the bodies were deposited over a short period.

Wiechmann, Harbeck and Grupe (2010) were interested in these skeletons as possible plague victims. Out of the 33 skeletons examined, Yersinia pestis DNA was detected in 10 individuals. A 30% positive result for aDNA testing of a site is I think as high has I’ve read before. Six positive skeletons were selected for further study with an additional four primer pairs for the Y. pestis plasmid pPCP1, including testing for the plasminogen activator and pesticin (a bacteriocin). All of the new markers were detected for at least one skeleton. From this data they conclude that the skeletons found in the mass grave represented plague victims.

Tran, Raoult, & Drancourt (2011) sent a letter to the editor to appear in the May issue of Emerging Infectious Diseases to contest a claim that Wiechman, Harbeck and Grupe didn’t make! The original letter describing these findings made no claim for which biovar of Yersinia pestis is present (Wiechmann, Harbeck, & Grupe, 2010 & 2011). Tran, Raoult and Drancourt (2011) appear to be anticipating that they will find that this strain does not belong to the oreintalis biovar, which would conflict with their previous assertions that all medieval Y. pestis they have found belong to the orientalis biovar. The wrangling over the biovars  and lineages of medieval Yersinia pestis is just beginning.

Historically what matters is that 75 people, mostly victims of Yersinia pestis, were buried beneath the Church of St Leonard in the Late Middle Ages. Of the six people tested by Wiechmann, Harbeck and Grupe (2010), there were 2 females and 4 males between ages of 8 and about 25. They were not clergy. If this is a representative selection of individuals, they probably were not high status townspeople based on their ages. The lineage of their Yersinia pestis is not the only unsolved mystery about these people left to be solved.

ResearchBlogging.org

Wiechmann I, Harbeck M, & Grupe G (2010). Yersinia pestis DNA sequences in late medieval skeletal finds, Bavaria. Emerging infectious diseases, 16 (11), 1806-7 PMID: 21029555

Tran T-N-N, Raoult D, Drancourt M. Yersinia pestis DNA sequences in late medieval skeletal finds, Bavaria [letter]. Emerg Infect Dis [serial on the Internet]. 2011 May [cited April 2011]. http://www.cdc.gov/EID/content/17/5/955.htm DOI: 10.3201/eid1705.101777

Wiechmann I, Harbeck M, Grupe G. Yersinia pestis DNA sequences in late medieval skeletal finds, Bavaria [letter]. Emerg Infect Dis [serial on the Internet]. 2011 May [cited April 2011]. http://www.cdc.gov/EID/content/17/5/955.htm  DOI: 10.3201/eid1705.102013

Plague DNA from Late Antique Bavaria

ResearchBlogging.org

The first plague pandemic was not recorded in Bavaria, or anywhere in the Germanic territory that I am aware of. The grave was not a typical ‘plague pit’. It was a rich grave of an adult woman and a young girl (individuals 166 and 167) from a cemetery in Aschheim, Bavaria. With no visible signs of illness or trauma, the remains were sent for a full genetic workup where they a revealed big surprise.

Ingrid Wiechmann extracted DNA from several teeth from each skeleton. She confirmed their gender by amplifying the amelogenin gene, and failure to amplify the Y specific marker. They were unable to prove that these individuals are mother and child because nuclear DNA was inconsistently amplified from the child. The mitochondrial DNA did amplify well and both yielded group 2A–C, haplotype 52, suggesting that they could be from the same matrilinear group.

So far, all of this was basically expected, but then Wiechmann amplified a section of the Yersinia pestis pla gene from each skeleton.  The plague testing was a stab in the dark based purely on the sixth century date for the remains and lack of other discernible causes of death. This should be a game changer.

This study is very important for plague research, particularly for first pandemic research, for three reasons. It provided timely support for Y. pestis as the agent of the medieval plagues in Europe. Yersinia skeptics had been growing quite loud and insistent lately, some even going so far as to claim that the agent in northern Europe was different than around the Mediterranean! Finding DNA from the first pandemic period also shows that it should be possible to extract plague DNA from the entire period of the late antique  and medieval plagues. Second, recovery of plague DNA came from a region where plague had never been previously reported in contemporary documents or by modern testing. When you look at the map below and realize that there are no plague reports from Bavaria or anywhere east of it, you realize how small and coastal our contemporary documentary record is for the first pandemic.

Mapping the first pandemic beyond the European coast and former Roman province is really in the court of the physical anthropologists. The documentary evidence for the first pandemic is really slim. This is a problem that archaeologists can solve if they begin routinely screening period graves. Immunological screening by the new immunochromographic (dipstick-like) test should be cheaper and more sensitive (Pusch et al, 2004 & Bianucci et al, 2008). This test can probably be done by archaeologists themselves. Extracting DNA from these remains will also be important in determining how many clones caused the waves of the first pandemic, and that will tell us how many times plague was introduced to Europe.

The third reason this find is so important is that it was found in a very normal, high status grave. This woman and child were carefully buried with “conspicuous grave goods” including a pair of bow fibulae one of which had a runic inscription on the back. These bow fibulae are considered to be high status goods in sixth century southwestern Germany reflected in this grave by leather coverings for them on the belt of each individual. Wiechmann and Grupe suggest that these non-locally made items may have been inherited. Finding plague in such carefully prepared graves in a cemetery that showed no “hints of mass infection or epidemic” suggests that plague victims could be in any period grave without obvious other cause of death. It seems likely that plague victims are being overlooked in many sixth and seventh century cemeteries.

Reference:

Wiechmann I, & Grupe G (2005). Detection of Yersinia pestis DNA in two early medieval skeletal finds from Aschheim (Upper Bavaria, 6th century A.D.). American journal of physical anthropology, 126 (1), 48-55 PMID: 15386257

Bianucci, R., Rahalison, L., Massa, E., Peluso, A., Ferroglio, E., & Signoli, M. (2008). Technical note: A rapid diagnostic test detects plague in ancient human remains: An example of the interaction between archeological and biological approaches (southeastern France, 16th–18th centuries) American Journal of Physical Anthropology, 136 (3), 361-367 DOI: 10.1002/ajpa.20818

Pusch CM, Rahalison L, Blin N, Nicholson GJ, & Czarnetzki A (2004). Yersinial F1 antigen and the cause of Black Death. The Lancet infectious diseases, 4 (8), 484-5 PMID: 15288817