GINO FORNACIARI
Department of Oncology
Paleopathology Laboratory
University of Pisa, Italy

Despite current opinion, the series of mummies in Italy, and in particular the single mummies, are quite numerous (Di Colo 1910; Terribile and Corrain 1986; Fulcheri 1991; Fornaciari and Capasso 1996). According to a recent survey there are 315 preserved bodies of Saints, including at least 25 mummies (Fulcheri 1996). The mummies are distributed over the whole Italian territory from the north (Aufderheide and Aufderheide 1991) to the south (Fornaciari and Gamba 1993), where it is possible to find the most important collections. The burials, which represent extremely precious paleopathological material, range from the medieval period, to the Renaissance, to more recent times, principally between the 17th and 19th centuries (Table 1). The samples vary from small numbers to several thousands of individuals as for example the Catacombs of the Capuchins of Palermo (Di Colo 1910).
The mummies in Italy can be divided into two different typological groups: natural and artificial mummies. The natural mummies, such as the mummy of Santa Zita in Lucca, Tuscany (Fornaciari et al. 1989h), or of Pandolfo Malatesta, prince of Fano (central Italy) (Fornaciari and Torino 1995), form the most consistent part of the Italian collections and were preserved for climatic and environmental reasons, without direct human intervention. On the other hand the artificial mummies, such as the royal mummies of the Aragonese Mausoleum of San Domenico Maggiore in Naples (Fornaciari 1986), were exclusive of particular members of society, either of high social classes or of great importance for the community, as for example kings or saints. With regard to the first category, there is sometimes documentary evidence of the special treatment to which the bodies were submitted, in order to facilitate the drying process and preservation. Natural mummification was facilitated by particular drainage methods of the cadaveric fluids, which were carried out in the crypts of the
churches, especially in southern Italy. Up until the past century, after the death of an important personage, the body was placed and left for several months in the vault of a church, probably in a seated position, with the cadaveric sewage collected in large stone or pottery vases named “cantarelle”. Even today, in Naples, the expression “drain off !” is an omen of death still addressed to people (Fornaciari 1984a; 1986). After a few months the body, still flexible but no longer draining, was laid horizontally in special tubs covered with volcanic soil, rich in minerals, which completed the dehydration and mummification processes. The corpse, by then completely desiccated and mummified, was dressed and positioned in its coffin (Fornaciari and Gamba 1993).
We shall now describe the mummies of the church of Santa Maria della Grazia, at Comiso (Sicily), and those of the Abbey of San Domenico Maggiore in Naples, which are the two series which have been studied more exhaustively.

THE CHURCH OF SANTA MARIA DELLA GRAZIA IN COMISO, SICILY
(18th-19th century)
The “Chapel of the Dead”, a mortuary chapel annexed to the church of Santa Maria della Grazia in Comiso at the beginning of the 18th century, contains 50 mummified bodies placed in niches above the entrance and in the right and lateral walls, at the sides (Fornaciari and Gamba 1993). Nearly all the bodies, which lie in an oblique position, with their faces always directed towards the inside of the church, belong to males whose age varies from young to adult to old. The individuals are Capuchin friars or laymen of the Third Order of the Capuchins. Twenty mummies are labelled with the individual’s name and date of death, ranging between 1742 and 1838. The mummies without a label are probably more ancient, dating back to 1693, when the church was erected. All the mummies were wearing monastic clothes, except for one, dressed in characteristic 18th century civilian clothes, which was placed vertically in the middle of the left wall. Examination of the bodies revealed that the mummies were natural, in the sense that none of the individuals had been submitted to any type of treatment, either evisceration or craniotomy. Natural mummification of the bodies was probably made possible by the hot and dry climate of Comiso, located at the same latitude as Tunis. Confirmation that the bodies in the chapel were dressed only after mummification derives from the discovery that all the monastic clothes presented a posterior longitudinal cut running from the bottom to the hood, and this cut must have been made to facilitate the dressing of a dry and therefore very stiff human body.
From the paleopathological point of view, the provincial Bernardino del Comiso, who died in 1742 at the approximate age of fifty, was affected by severe arteriosclerosis with calcifications of the lumbar aorta and iliac arteries; a 35 year old individual presents an enormous enlargement of the thyroid gland, histologically a colloid goiter (Fornaciari et al. 1994b); another, of about 45 years of age, suffered from massive splenomegaly with infarctual areas; a man aged 30-35 shows pulmonary fibrosis with multiple, apical calcifications, and was probably affected by tuberculosis; another individual, who died at about 60 years of age, displays diffuse skin acariasis, treated “in vita” by sulphur ointments, with hyperkeratosis and abundant eggs, nymphs and mites at different stages of development still in situ. There are also a case of colon dìverticulosis, two cases of inguino-scrotal hernia and a case of varicose veins, with ulcers, of the lower limbs (Fornaciari and Gamba 1993).

THE BASILICA OF SAN DOMENICO MAGGIORE IN NAPLES
(15th-16th century)
The Basilica of San Domenico Maggiore, which dates back to the beginning of the 14th century, is one of the largest and most important churches in Naples. The humanist
Giovanni Pontano and the philosophers Tommaso Campanella and Giordano Bruno studied in this Abbey. Saint Thomas Aquinas taught in the annexed convent of the Dominicans. The impressive Sacristy of San Domenico Maggiore, in a suspended gateway close to the vault, contains 38 wooden coffins, or “arks”, with the bodies of 10 Aragonese princes and other Neapolitan nobles, who died in the 15th and 16th centuries. The coffins were initially placed in different parts of the church, but in 1594 Philip II, King of Spain, ordered that they should be gathered in the Sacristy (Miele 1977). The sarcophagi, richly dressed in precious clothes made of silk, brocade and other material, are distributed in two rows, one above the other. The smaller coffins of the lower row are generally of anonymous individuals, while the larger coffins of the upper row are identified by the coats-of-arms and the names of the personages buried inside. In particular they include the Aragonese kings Alfonso I (who died in 1458), Ferrante I (1494), Ferrante Il (1496), Queen Giovanna IV (1518) and the Marquis of Pescara Ferdinando d’Avalos, who won the famous battle of Pavia against the French King François I in 1525.
A first survey showed that the bodies contained in the coffins were generally mummified and well preserved. That of Pietro of Aragon, III Duke of Montalto, who died in 1552 at the age of 12 (Fornaciari 1984b), is an example of a typical burial of Santa Maria Maggiore. It consists in a large external sarcophagus (or “ark’), containing a second coarse wooden coffin; there are plants, such as laurel, rosemary and box covering the richly dressed body in 16th century style, which lies on a layer of small lime fragments, for the drainage of the cadaveric fluids. The mummy, with the hands crossed on the pubis, resulted in an excellent state of preservation.
The mummies of San Domenico Maggiore are unique in Italy not only for the antiquity and excellent state of preservation of the bodies, but also for the fame of the personages, whose lives and causes of death are well known. King Ferrante II, for example, died of malaria, while the Marquis of Pescara died of pulmonary tuberculosis. The possibility of comparing the paleopathological with the historical data provided extremely interesting results. Up to now, the only mummies of this type known in Europe have been those of the Hapsburg princes in Vienna (Kleiss 1977).
The sarcophagi of San Domenico Maggiore were carefully examined from 1984 to 1987 by a team of the Institute of Pathology of Pisa University. Radiography followed by anthropological and autoptic examination on site were carried out on the mummies; the laboratory studies were performed in Pisa. The extremely precious clothes and the jewels of the buried individuals have been carefully recovered and will soon be restored and displayed at the Museum of Capodimonte by the Superintendence of Naples.
Let us now summarise the results obtained so far: of the 38 sarcophagi explored, 8 were found to be empty, while one contained a double deposition. All depositions resulted to be more or less disturbed. There were 27 primary depositions and 4 secondary depositions, or redepositions. Fifteen individuals, equal to 48.4% of the totality, had been submitted to embalming, while 12, equal to 38.7%, had not been treated (Table 2). Therefore, the majority of the individuals had been embalmed and this is certainly not surprising, considering the high social class of the individuals buried in San Domenico. From the physician Ulisse Aldrovandi we know that during the Renaissance “the European kings and great personages used to entrust embalming of their bodies to their doctors and surgeons” (Aldrovandi 1602). With regard to San Domenico, a document informs us about the embalming of the body of Antonio of Aragon, IV Duke of Montalto, who died in 1584: “Lo corpo e in sacrestia imbalsamato” (his body embalmed in the sacristy) (Vultaggio 1984). The skeletonized individuals were 6, corresponding to 22.2% of the totality; however, there was only one case of skeletonization, corresponding to 6.7%, among the embalmed individuals (Table 2).
For the partial preservation of the bodies the embalming process was obviously very important . In this respect, 9 individuals out of 14, including 5 children and 4 adults, had been eviscerated by a long anterior incision running from the neck basis to the pubic symphisis (Table 3). In order to penetrate the thorax, the sternum was cut or sewn or even removed; otherwise, the ribs or costal cartilages were cut sideways with the aid of shears; in one particular case both operations were performed. The sternum was generally cut in new-born babies and sucklings, and less often in adults where, as in modern autopsies, the costal cartilages were cut on each side of the sternum and the anterior thoracic wall was removed. The other 5 individuals, which were 1 child and 4 adults, only presented an abdominal incision running from the xyphoid process of the sternum to the pubis (Table 3). In this case the thorax was necessarily eviscerated through the diaphragm. Of the 15 cases of embalmed individuals 13 had the brain removed by craniotomy, which was horizontal and circular in 7 cases, or posterior and often circular, in 6 cases (Table 4). Widespread unfleshing of the muscular masses was observed at the level of the dorsum, the glutei, and the limbs in 4 cases, which included 2 new-born babies and 2 adults. The embalming material consisted principally of resinous substances, which were present in 10 cases; wool or similar material, and clay or earthy substances, were present in 6 cases; lime was found in 4 cases, leaves or twigs in 3 cases, while tow, sponges and mercury were only used twice (Table 5). Finally, in 4 cases, including 2 new-born babies and 2 adults, the body was wrapped in bandages previously soaked in resinous substances. Wood for the inner coffins was determined in 16 cases of primary depositions; there was a high incidence of pine-wood, used in 9 depositions, followed by poplar in 4 depositions and chestnut wood in 3 (Table 6). Resinous essences, which were thought to favour the preservation of the bodies, were also used in the case of the coffins. These very complex evisceration and embalming methods indicate long-practised and diffused customs but, as already said, some well preserved individuals show no apparent signs of embalming. Natural mummification of the bodies can probably be attributed to the warm and dry climate of Naples during the summer. Preservation of the bodies may have been favoured by the particular microclimatic conditions of San Domenico Maggiore, as well as by the disposition of the coffins, placed near the windows of the sacristy, at about 5 m of height. Furthermore, two large rooms, certainly devoted to dehydration of the bodies, have been discovered recently in the crypts of San Domenico. These two rooms were provided with numbered spaces for the coffins, with wide ventilation shafts, and with thick sand beds, for the gathering of the cadaveric fluids. The mummies of San Domenico are likely to have been submitted to the previously described treatment of natural mummification practised in Naples until the last century.
We shall now describe two cases of infectious disease and two cases of neoplastic pathology, from a strictly paleopathological point of view.
The mummy of an anonymous 2-year-old boy, whose death dates back to the mid 16th century (radiocarbon dating is 1569± 60), presented a diffuse vesiculo-pustular exanthema type eruption. Macroscopic aspects and regional distribution suggested a case of smallpox. This possibility was confirmed by light microscopy and indirect immunofluorescence with anti-vaccine-virus antibody. Electron microscopy revealed, among the residual bands of collagen fibres, pyknotic nuclei, and membrane remains with rare desmosomes, many egg-shaped, dense virus-like particles (250 x 50 nm), composed of a central dense region (or core) surrounded by a low density area. Following incubation with human anti-vaccinia-virus antiserum, after protein A-gold complex immunostaining, the particles were completely covered by protein A-gold. These results showed that the antigenic structure of the viral particles was well preserved and that this Neapolitan child died of a severe form of smallpox some four centuries ago (Fornaciari and Marchetti 1986a, 1986b, 1986c, 1986d, 1986e; Fornaciari et al. 1989i).
The study of a case of treponematosis in the mummy of Maria of Aragon (1503-1568), Marquess of Vasto in southern Italy, appeared to be particularly interesting.
Famous for her beauty, this noblewoman of the Italian Renaissance belonged to the intellectual and religious circles of lschia, which also included a friend of Michelangelo’s, the poetess Vittoria Colonna. An oval 15×10 mm cutaneous ulcer covered by a linen dressing with ivy leaves appeared on the left arm of the mummy. lndirect immunofluorescence with human anti-treponema pallidum antibody identified a large number of filaments with strong yellow-green fluorescence and the morphological characteristics of fluorescent treponemes. Morphological aspects typical of the spirochetes, as for example the axial fibril, were evidenced by the ultrastructural study. lmmunohistochemical and ultrastructural findings clearly demonstrated treponemal infection and the cutaneous ulcer resulted to be typical of third-stage luetic gumma. Venereal syphilis was the most probable diagnosis (Fornaciari et al. 1989c, 1989d, 1989e, 1994a). This discovery is extremely important since it dates back to the 16th century and can help to clarify the biology of treponema in the epidemic phase of the disease.
Another important paleopathological case is that of Ferdinando Orsini, Duke of Gravina in Apulia, who died in 1549. The mummy presents wide erosion of the upper orbital margin and the glabella, and complete destruction of the right nasal and retro-orbital bones. Histology showed solid neoplasia, with cords of spindle-shaped cells destroying compact and spongy bone and forming osseous lacunae, with no bone reaction. A widely destroying skin epithelium seems to be the most probable diagnosis (Fornaciari et al. 1989b).
The artificial mummy of Ferrante I of Aragon, King of Naples, who died in 1494 at 63 years of age, was submitted to autopsy which revealed in the small pelvis a fragment of a cave muscular organ, which reached the dimensions of 6x4x1cm after dehydration. Histologically, neoplastic epithelial cells disposed in cords, nests and glands were disseminated in a fibrous stroma containing scattered striated muscular fibres. The cells were tall, crowded, with abundant cytoplasm and pseudo-stratified pleomorphic hyperchromatic nuclei. The scarce mucus was limited to the pseudoglandular formations, as appeared from the specific Alcian-blue staining. The use of a monoclonal antibody versus pancytokeratin displayed strong intracytoplasmic immunoreactivity of the tumoral cells. The ultrastructural study evidenced well preserved pleomorphic nuclei with indented membranes. These results clearly indicate a mucinous adenocarcinoma infiltrating the muscular-fibrous layers of the small pelvis. However, the site of the primary neoplasm was at first impossible to establish: the histological features only suggested prostatic adenocarcinoma or an adenocarcinoma of the digestive tract (Fornaciari et al. 1993). Since colorectal tumours are characterised by frequent mutations of the K-ras oncogene – whereas in prostatic adenocarcinomas such mutations are extremely rare – we decided to investigate the status of the K-ras gene in the DNA tumour extracted from the mummified tumour tissue (Caligo et al. 1994). The samples were subjected to a nested PCR protocol designed to yield a 77 bp K-ras fragment encompassing codon 12, the main hotspot for mutations in colon cancer. The hybridization with 32P-labelled mutation specific oligonucleotide probes showed the presence, in the tumour sample, of a K-ras codon 12 point mutation: the normal sequence GGT (glycine) was altered to GAT (aspartic acid). This data clearly demonstrates that Ferrante I was affected by a cancer of the digestive tract, most probably a colon adenocarcinoma (Marchetti et al. 1996). The observed genetic change indeed represents the most frequent mutation of the K-ras gene in present-day colorectal cancer and it is thought to be induced by alkylating agents (Topal 1988). Therefore we speculate that mutagens similar to those responsible for the induction of K-ras mutations in contemporary colon cancer could be present in the rich diet of a court of the XV century, as supported by paleonutritional studies (Fornaciari et al. 1989f). This is the first time an oncogene mutation has been evidenced in an ancient tumour.
In conclusion, the paleopathological study of the mummies of eleven adult individuals from the Abbey of San Domenico Maggiore, with good or excellent preservation, allowed us to diagnose two cases of cancer. The number of available specimens was very limited ; however, it was possible to observe an incidence of neoplastic pathology (1 8.8%), similar to the one we find nowadays.
Among the “minor” pathologies we wish to point out the mortal stab-wound, with surrounding hemorragic infarction, between the eighth and the ninth left rib in the mummy of an anonymous gentleman who died at the age of about 27 in the second half of the 16th century (Fornaciari et al. 1989a); a case of liver cirrhosis with typical nodules; a case of acute oedema, or pneumonia, in an individual with severe anthracosis; an individual affected by calculosis of the gall-bladder with chronic colicystitis (Fornaciari et al. 1989g).
Very important is the recent discovery of a sequence of hepatitis E virus (HEV) in the DNA extracted from the mummy of Maria of Aragon (Marota et al. 1996)
The majority of Italian mummies have not yet been examined, but future researches in
this field will certainly provide extremely important results for the history of diseases.

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TABLES

SITE N. TYPE CENTURY

VENZONE 15 NATURAL 14th-18th
(Northern Italy)
URBANIA 18 NATURAL 17th-19th
(Central Italy)
MEDICI MAUSOLEUM 39 NATURAL & 16th-18th
(S.Lorenzo, Florence) ARTIFICIAL
FERENTILLO 16 NATURAL 18th-19th
(Central Italy)
ROCCA DI PAPA hundreds NATURAL 16th-18th
(Central Italy)
NAVELLI hundreds NATURAL 13th-19th
(Central Italy)
ARAGONESE MAUS. 31 NATURAL & 15th-19th
(S.Domenico, Naples) ARTIFICIAL
ALTAVILLA IRPINA hundreds NATURAL 18th-19th
(southern Italy)
SAVOCA tens NATURAL 17th-18th
(Sicily)
Comiso 50 NATURAL 18th-19th
(Sicily)
CAPUCHINS CATAC. thousands NATURAL 16th-20th
(Palermo)

Table 1. Collections of mummies in Italy.

TOTAL No. INDIVIDUALS 31
___________________________
PRIMARY DEPOSITIONS 27 (87.1%)
SECONDARY DEPOSITIONS 4 (12.9%)
NATURAL MUMMIES 12 (38.7%) SKELETONIZED 5 (41.7%)
ARTIFICIAL MUMMIES 15 (48.4%) SKELETONIZED 1 ( 6.7%)

Table 2. Basilica of S. Domenico Maggiore (Naples): general situation
of the depositions.

JUGULO-PUBIC INCISION 9
XIPHO-PUBIC INCISION 5
UMBILICAL-TRANSVERSE 4
(associate with the above)

Table 3. Basilica of S. Domenico Maggiore (Naples): types of evisceration
in artificial mummies.

NUMBER OF MUMMIES 15
_______________________________

HORIZONTAL CRANIOTOMY 7
POSTERIOR CRANIOTOMY 6
WITHOUT CRANIOTOMY 2

Table 4. Basilica of S. Domenico Maggiore (Naples): types of craniotomy in
artificial mummies.

RESINS WOOL SOIL LIME LEAVES TOW SPONGES MERCURY COTTON
OR OR OR 
CLAY ASH TWIGS
10 6 6 4 3 2 2 2 1
66.7% 40.0% 40.0% 26.7% 20.0% 13.3% 13.3% 13.3% 6.7%

Table 5. Basilica of S. Domenico Maggiore (Naples): frequency of different
embalming materials in artificial mummies.

NATURAL MUMMIES ARTIFICIAL MUMMIES TOTAL
FIR 5 4 9 (56.2%)
POPLAR 2 2 4 (25.0%)
CHESTNUT 0 3 3 (18,7%)

Table 6. Basilica of S. Domenico Maggiore (Naples): types of wood used
for inner coffins in primary depositions.