Osteological and molecular identification of brucellosis in ancient Butrint, Albania

Abstract

Ancient skeletal remains can harbor unique information about past civilizations at both the morphological and molecular levels. For instance, a number of diseases manifest in bone, some of which have been confirmed through DNA analysis, verifying their presence in ancient populations. In this study, anthropological analysis of skeletal remains from the ancient Albanian city of Butrint identified individuals with severe circular lytic lesions on their thoracic and lumbar vertebrae. Differential diagnosis suggested that the lesions resulted from pathologies known to affect these skeletal regions, such as tuberculosis (TB) or brucellosis. Relevant bones of two adolescent males from the 10th to 13th century AD that displayed the lesions, along with unaffected individuals, were collected in the field. Genetic screening of the skeletal samples for TB was repeatedly negative, thus additional testing for Brucella spp.-bacteria of livestock and the causative agent of brucellosis in humans-was conducted. Two Brucella DNA markers, the IS6501 insertion element and Bcsp31 gene, amplified from the affected vertebrae and/or ribs, whereas all unaffected individuals and control samples were negative. Subsequent DNA sequencing confirmed the presence of the brucellar IS6501 insertion element. On the basis of the skeletal lesions, negative tests for TB, and positive Brucella findings, we report a confirmed occurrence of brucellosis in archaeologically recovered human bone. These findings suggest that brucellosis has been endemic to the area since at least the Middle Ages.

Skeletal evidence of brucellosis in a medicolegal context: A report of two cases

“…The lumbar section of the spine is affected in 80% of the cases [1]. The inflammatory brucellar course in the vertebral column initially affects the intervertebral disc tissues and the bone of the antero-superior corner of the body immediately below the affected disc [1,7,29]. Subsequently the fibres of the anterior longitudinal ligament are affected, as is the periosteum of the anterior face of the vertebral body [1].…”

“…Direct person-to-person spread is extremely rare [6]. An incubation period ranging from three weeks to several months is required before the onset of the disease [7].…”

“…Primary infection typically manifests as chronic respiratory illness, fever and splenomegaly [7]. Secondary infection of skeletal tissue can occur when the bacteria become systemic and spread to cancellous bone [7].…”

Brucellosis in Immunocompromised Hosts

“…The occurrence of brucellosis in archaeologically recovered human bones, belonging to 2 young patients who lived in Albania between the 10th and 13th centuries, has been confirmed using DNA sequencing technology [96]. Also, 6 old cases of brucellosis with skeletal involvement belonging to the period between the 5th and 12th centuries have been recovered from Western Europe Mediterranean countries using molecular techniques such as PCR.…”

Evidence of tuberculosis among children in medieval (13th–15th century) Wrocław: A case study of hip joint tuberculosis in a juvenile skeleton excavated from the crypt of the St. Elizabeth church

“…Wimberger's sign), metaphyseal serration, sawtooth -like appearance of the metaphysis; mild to severe periosteal reactions (Rasool and Govender, 1989), eg. lamellar periostitis within the anterior surface of the tibiae (sabre shins) Brucellosis Sacro-illiac joints and lumbar vertebrae (Mutolo et al, 2012;D'Anastasio, et al, 2011)large peripheral joints (hip and knee more frequently affected than sacroiliac joint and spine) (Alshaalan et al, 2014) sacroiliitis or lumbar spondylitis (D'Anastasio et al, 2011); osteomyelitis in long bones and the pelvis, arthritis (Kelly et al, 1960;Ariza et al, 1985) …”