Neonate Human Remains: A Window of Opportunity to the Molecular Study of Ancient Syphilis

Abstract

Ancient DNA (aDNA) analysis can be a useful tool in bacterial disease diagnosis in human remains. However, while the recovery of Mycobacterium spp. has been widely successful, several authors report unsuccessful results regarding ancient treponemal DNA, casting doubts on the usefulness of this technique for the diagnosis of ancient syphilis. Here, we present results from an analysis of four newborn specimens recovered from the crypt of “La Ermita de la Soledad” (XVI–XVII centuries), located in the province of Huelva in the southwest of Spain. We extracted and analyzed aDNA in three independent laboratories, following specific procedures generally practiced in the aDNA field, including cloning of the amplified DNA fragments and sequencing of several clones. This is the most ancient case, reported to date, from which detection of DNA from T. pallidum subspecies pallidum has been successful in more than one individual, and we put forward a hypothesis to explain this result, taking into account the course of the disease in neonate individuals.

Ancient pathogens in museal dry bone specimens: analysis of paleocytology and aDNA

“…The identification of treponemal aDNA in the case of adult treponemal disease by Kolman et al [40] could not be repeated. Nevertheless, Montiel et al [48] succeeded in detecting treponemal aDNA from archaeological neonate human remains. They argue that congenital syphilis produces lethal effects to the fetus or newborn rapidly so that the level of spirochetes is high enough for the detection of aDNA.…”

“…Kolman et al [40] present the only positive aDNA detection in a case of venereal syphilis from Easter Island. Montiel et al [48] detected treponemal aDNA in congenitally infected neonate human remains. Failed attempts to identify ancient treponemal DNA sequences were conducted by Bouwman and Brown [11], Barnes and Thomas [7], and von Hunnius et al [84].…”

“…The primers F284 (5′-GTGATCCTCTGTCATCCCCG-3′) and R370 (5′-TCCACCTCACGAGACAAAGG-3′) from Montiel et al[48] were used to amplify a fragment of the acidic repeat protein gene (arp). According to Montiel et al[48] und Kolman et al[40] amplification was conducted in a 25 µL reaction with GeneAmp buffer (PE Applied Biosystems, Script SK under assistance of the PCR-Script Cloning Kit and cloned in XL1-Blue MRF' cells (Stratagene, La Jolla). M13 or PCR primers were used for sequencing with a Sequencing Kit FS version (PE Applied Biosystems) and a Perkin-Elmer 373 DNA Sequencer.…”

Ancient bacterial genomes reveal a formerly unknown diversity ofTreponema pallidumstrains in early modern Europe

“…Most notably, the bones that likely contain a large amount of treponemal agents belong to congenitally infected neonates. These fragile remains rarely survive and are, even when present, often overlooked in the archaeological record 44,45 . Previously, it would not have been feasible to use samples with low bacterial loads to detect pathogen DNA; however, recent advances in target-enrichment, high-throughput sequencing and sensitive screening methods for aDNA have aided overcoming this issue 46 .…”

Molecular analysis of ancient caries

“…The diseases that have been molecularly studied so far are those believed to have caused the majority of the documented human epidemics, such as tuberculosis (Mycobacterium tuberculosis) [1,2], plague (Yersinia pestis) [3,4], leprosy (Mycobacterium leprae) [5,6] and syphilis (Treponema pallidum pallidum) [7,8], among others [9][10][11]. However, there are other microorganisms, such as bacteria from the dental plaque, that have accompanied humans since remote times for which their history is still not well established, and that could help us to understand prehistoric populations in depth.…”