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]

Lice, Ancient DNA, and Napoleon’s Grand Army

Identification of human body lice. (Raoult et al, 2006)

Identification of human body lice. (Raoult et al, 2006)

Life in Napoleon’s Grand Army wasn’t always so grand. The Russian campaign was a disaster, recently most tangibly manifest in the mass grave found at Vilnius, Lithuania, in 2001. Local records suggested that the remains belonged to Napoleon’s soldiers who paused at Vilnius during their retreat from Moscow in 1812. The densely packed bodies were buried at the same time leaving behind buttons, buckles and gear of 40 regiments of Napoleon’s army. The initial trench revealed 717 skeletons at 7 corpses per meter squared, predicting 2000-3000 corpses at the site. Winter weather may have claimed most of the soldiers, but they were also known to have been plagued by lice and fevers.

For Raoult et al (2006) this was the perfect test case for detecting lice and louse-borne diseases in archaeological remains. After perfecting their technique of isolating cultured, dried human lice (Pediculus humanus humanus) in the laboratory*, they obtained soil from the mass grave at Vilnius. They isolated fragments of five lice from two kilograms of soil containing remains of bones and clothing, and confirmed the identification by “binocular magnification” (right), scanning electron microscopy and isolated P. humanus humanus DNA to confirm the species. Three of the lice produced Bartonella quintana DNA (trench fever) but none of the lice yielded Rickettsia powazekii DNA (typhus) or Borellia recurrentis DNA (relapsing fever).

They extracted DNA from 72 unerupted teeth from 35 skeletons for ‘suicide’ PCR pathogen detection. They found R. powazekii (typhus) in teeth from three skeletons and B. quintana (trench fever) teeth from seven skeletons. Overall, louse-transmitted disease was identified in 10 out of 35 skeletons (28.6%). B. quintana is believed to be the most common louse-transmitted pathogen in Antiquity and the Middle Ages. Trench fever alone is usually not fatal but when combined with the harsh conditions on the retreat from Russia it could have been a significant contributing factor. Epidemic typhus on the other hand is a more virulent organism and is more likely to be the cause of death of the three individuals in whose teeth R. powazekii was discovered.

The ability to identify pathogen vectors, in this case the human louse, and amplify pathogen DNA from both ancient vectors and humans is a big advance in our understanding of infectious disease in pre-modern populations.

* I don’t envy the research assistants or students who had to maintain the colonies of human lice in the lab, or roast them incubators to simulate conditions in soil.

ResearchBlogging.org
Raoult D, Dutour O, Houhamdi L, Jankauskas R, Fournier PE, Ardagna Y, Drancourt M, Signoli M, La VD, Macia Y, & Aboudharam G (2006). Evidence for louse-transmitted diseases in soldiers of Napoleon’s Grand Army in Vilnius. The Journal of infectious diseases, 193 (1), 112-20 PMID: 16323139

Trench Fever and Plague in 14th Century France

Five Skeletons from Bondy, France. (Source: Tran et al, 2011a)

The Marseille plague group has been suggesting for some time now that human lice could be a major vector of medieval plague. To test their hypothesis the group devised a multiplex PCR screening method to rapidly screen many aDNA samples for seven pathogens that could cause medieval epidemics, including relapsing fever and trench fever transmitted by human lice. Their method was previously published for a  screening of mass burials from medieval Venice (Tran et al, 2011b).

In their most recent publication, they have examined 14 teeth from five skeletons found in Bondy, near Paris, France (Tran et al, 2011a). The remains are from people of varied ages and both genders, radiocarbon dated to the 14th century (1297-1373 with 70% probability). They were also screened for the pathogens that cause anthrax, trench fever, relapsing fever, typhus, typhoid fever, smallpox and plague. As in Venice, they only found two of these, trench fever (Bartonella quintana) and plague (Yersinia pestis) (Tran et al, 2011a; Tran et al, 2011b). Overall, B. quintana was found in three individuals and Y. pestis in two individuals, with one individual (#73, far left skeleton half revealed in the picture) positive for both. Trench fever is transmitted by the human louse and Tran et al  (2011a) see this as supporting their hypothesis that lice transmitted the plague as well. While one unfortunate co-infected individual is not proof, it does add one more piece of evidence to support their hypothesis. Plague was confirmed by sequencing and identified as the orientalis biovar. This group has previously asserted that the orientalis biovar is transmitted much more efficiently by lice than other Y. pestis biovars (Avyadurai et al, 2010).

They are citing one more paper as ‘in press’ in Tran et al, 2011a. It looks like there is a summary paper coming up that will lay out all their arguments for lice as transmitters of medieval plague, so stay tuned!

ResearchBlogging.org

Tran TN, Forestier CL, Drancourt M, Raoult D, & Aboudharam G (2011a). Brief communication: Co-detection of Bartonella quintana and Yersinia pestis in an 11th-15th burial site in Bondy, France. American journal of physical anthropology PMID: 21541920

Tran TN, Signoli M, Fozzati L, Aboudharam G, Raoult D, & Drancourt M (2011b). High throughput, multiplexed pathogen detection authenticates plague waves in medieval Venice, Italy. PloS one, 6 (3) PMID: 21423736

Ayyadurai S, Sebbane F, Raoult D, & Drancourt M (2010). Body lice, yersinia pestis orientalis, and black death. Emerging infectious diseases, 16 (5), 892-3 PMID: 20409400