Genetic Studies in Paraguay: Blood Group, Red Cell, and Serum Genetic Patterns of the Guayaki and Ayore Indians, Mennonite Settlers, and Seven Other Indian Tribes of the Paraguayan Chaco 1

STEPHEN M. BROWN,2 D. CARLETON GAJDUSEK,3 WEBSTER C. LEYSHON,4 ARTHUR G. STEINBERG,5 KENNETH S. B R O W N 4

AND CYRIL C. CURTAIN 6

2 National Institute of Child Health and Human Development, N.I.H., Bethesda; present address: Program in Epidemiology, School of Public Health, University of California, Berkeley; 3 National Institute of Neuro-logic Diseases and Stroke, N.I.H., Bethesda; 4 Human Genetics Branch, National Institute for Dental Research, N.I.H., Bethesda; 5 Department of Biology, Case Western Reserve University, Cleveland; 6 Division of Animal Health, Commonwealth Scientific and Industrial Research Organization, Melbourne

KEY WORDS P a r a g u a y • S o u t h A m e r i c a n I n d i a n s • M e n n o n i t e s • B lood g r o u p s • S e r u m G l o b u l i n s • H a p t o g l o b i n s • H e m o g l o b i n s .

ABSTRACT G e n e t i c s t u d i e s of 5 4 0 P a r a g u a y a n I n d i a n s f r o m n i n e tr ibal g r o u p s a n d 51 M e n n o n i t e s are p r e s e n t e d f o r ABO, M N S s , P 1 ; Rh , Kell , L e w i s , D u f f y , D i e g o ; f o r s e r u m i m m u n o g l o b u l i n s a n d h a p t o g l o b i n s , G 6 P D - d e f i c i e n c y , a n d t h a l a s s e m i a trait.

Group O g e n e f r e q u e n c i e s f or al l I n d i a n g r o u p s w e r e 1 .00; f or r ( c d e ) , 0 . 0 0 . T a p i e t e , L e n g u a , T o b a , a n d S a n a p a n a R, ( C D E ) f r e q u e n c i e s w e r e a m o n g t h e

h i g h e s t ever reported . N f r e q u e n c i e s w e r e h i g h f o r A c h e K w e r a ( G u a y a k i ) , L e n g u a , Cherot i , Gua-

rayu , T a p i e t e ; N a n d s l o w f o r Ayore . M S f r e q u e n c i e s w e r e h i g h f o r S a n a p a n a , L e n g u a , Ayore; N s for T a p i e t e . D i e g o w a s n o t a b l y a b s e n t f or T o b a , L e n g u a , G u a r a y u , T a p i e t e , Ayore . H o m o g e n e o u s f r e q u e n c i e s f o r Fya ( 1 . 0 0 0 ) o c c u r r e d a m o n g G u a r a y u a n d T a p i e t e , a n d f o r P, a m o n g G u a y a k i . Inv(a) f r e q u e n c i e s w e r e l o w f o r Cherot i , C h u l u p i , G u a y a k i . Hp 1 a m o n g G u a y a k i ( A c h e K w e r a 0 . 1 5 ) i s l o w e s t e v e r reported .

G 6 P D d e f i c i e n c y a n d a b n o r m a l h e m o g l o b i n s w e r e u n i f o r m l y a b s e n t f r o m al l groups .

M e n n o n i t e r e s u l t s w e r e h o m o g e n e o u s a n d p o i n t t o w a r d D u t c h o r i g i n s . D i f f e r e n c e s a m o n g g r o u p s s tud ied , a n d b e t w e e n P a r a g u a y a n a n d o t h e r A m e r -

i n d s e m p h a s i z e i m p o r t a n c e of g e n e t i c dr i f t a n d f o u n d e r pr inc ip l e . A b a n d o n m e n t o f the ir tr ibes by m i x e d - b l o o d o f f s p r i n g i s part ly r e s p o n s i b l e f or a p p a r e n t g e n e t i c pur i ty a n d h o m o g e n e i t y o f groups .

THE PRESENT STUDY

Blood specimens from 540 representa-tives of nine Paraguayan Indian tribes and 51 Mennonite colonists in the Gran Chaco have been studied for blood group, serum, and red cell genetic factors. Data are pre-sented for ABO, MNSs, P„, Rh-Hr, Kell, Lewis, Duffy and Diego blood groups; for glucose-6-phosphate-dehydrogenase (G6PD)

A M . J . P H Y S . A N T H R O P . , 41: 3 1 7 - 3 4 4 .

deficiency and thalassemia trait; for Gm and Inv serum immunoglobulin markers, and for serum haptoglobins.

ETHNOGRAPHIC A N D BIOLOGIC BACKGROUND

Southeast Paraguay. The Paraguay ' Supported in part by Grant GM 07214 from the

National Institute of General Medical Sciences, Na-tional Institutes of Health, U.S.A.

317

3 1 8 S. BROWN, GAJDUSEK, LEYSHON, STEINBERG, K. BROWN A N D CURTAIN

River, in its southward course, divides Paraguay into halves. To the south and east of the river lie the rich forests in which the more densely populated areas of the country are found (fig. 1). Dry for most of the year, these forests are flooded during the brief summer rainy season. Here still roam two or three bands of Guayaki Indians, a hunting, honey-gather-ing group of cannibalistic, white-skinned people. Two of these bands, recently brought out of the forests and settled, are included in this study.

The Guayaki are of the Tupi-Guarani ethnolinguistic group, from which the mainstream of Paraguayan mestizo culture derives. Much has been written, mostly based on legend and hearsay and very brief encounters, about these fascinating, shy, stone-age, short-statured wanderers since their first contact with Jesuits in the eighteenth century. It is likely that Pedro Lozano, Jesuit historian, first contacted the Guayaki between 1717 and 1736 (Lozano, 1873), and describes another Jesuit lin-guist as having had three earlier contacts. From that time until they emerged from the forests in 1959 and settled in the region of San Juan Neopomuceno, their history is elusive. The Guayaki live amidst more ad-vanced Guarani tribes who have evolved into the agricultural sedentary mestizo people of Eastern Paraguay. During the eighteenth century, Guarani war parties were dispatched by the Jesuits to capture Guayaki for purposes of proselytization. More recently the Guarani mestizos have often raided Guayaki bands in reprisal for their killing of a horse or cow for meat, or as means of obtaining cheap domestic manpower. These raids have distorted the age and sex distribution of the Guayaki population.

Late in the nineteenth century, La Hitte and ten Kate twice visited and studied the Guayaki (La Hitte and ten Kate, 1897) and documented with photographs the Indians, their tools and a complete skeleton. There-after, intermittent contacts with the Guayaki have occurred until their recent settlement (Mayntzhusen, '25; Bertoni, '41).

Two "clans" or bands of Guayaki, the Ache Gatu and the Ache Kwera, collec-tively referred to as the "southern group"

by Metraux ( '46) , once roamed the region of Tayao, about 50 miles north of Villarica, west of the Parana River. They emerged and settled in 1959 and 1962, respectively. At least two other Guayaki bands inhabit the region north and east of Villarica, and at least two bands have been sighted fur-ther north, near the Brazilian border. Thus, the two bands of this study comprise from one-third to one-half of the total Guayaki population. The earlier settled Gatu tribe had been captured in the early 1950's, but escaped; the band numbered twenty at the time of its emergence in 1959, but others joined later. The Kwera group, who emerged in 1962, brought the total Guayaki population of the settlement to about 120, of which number at least 45 died of epidemic respiratory (primarily measles and influenza-like syndromes), and gastrointestinal infections during the first four years af ter their emergence.

The Guayaki are a polyandrous, highly promiscuous, inbred and incestuous group, with a paucity of younger females of child-bearing age, owing to their custom of female child sacrifice on the death of an adult, and the capture of young girls by local farmers. Cannibalism was practiced among the Ache Gatu, with a variety of ritual taboos, when death f rom any cause struck a member of the band. Because of the small band size and the relatively small number of young fecund females, the Guayaki show a dramatic occurrence of "founder" or "bottleneck" effect in which their successive gene pools are determined by very few individuals.

Demographically, the Guayaki group studied, exclusive of the 88 listed recently deceased individuals, can be summarized as follows:

Mean age Population (years) *

M F T M F T

Ache Kwera Study group

(al l ages ) 23 17 40 20 20 20 Adults * 11 11 22 33 29 31 Children * 12 6 18 8 3 6

Ache Gatu Study group

(a l l ages ) 14 8 22 25 35 29 Adults 9 6 15 37 43 39 Children 5 2 7 4 8 5

* Ages of subjects were estimated, and adults in-cluded all 5:13 years, children all < 13 years.

GENETIC STUDIES IN PARAGUAY 3 1 9

1 8 °

2 0 °

22°

2 4 °

L E G E N D o too zoo soo

• City or town Kilometers I m m m l I I • Bleeding sites M«.. I I U I I I II M T H 0 100 200 800

Fig. 1 Map of Paraguay, northern Argentina, and southeastern Bolivia, showing loca-tions of bleeding sites, Indian encampments, Mennonite colonies, and other place names mentioned in text.

S O U T H A M E R I C A

Piitrt* _ I Sastra Q \ Ptt«rf* _ Casado• Fltodclfla • • Lomo Plata

FERNHEIM YoUa Sanaa • |/ / La EiMron NEULAND I M

MENNONITE COLONIES

3 2 0 S. BROWN, GAJDUSEK, LEYSHON, STEINBERG, K. BROWN AND CURTAIN

Six Kwera and seven Gatu individuals were missing from the study, and our in-digenous informants reported 40 Kwera and 48 Gatu as dead. These figures bring to 86 and 77, respectively, the sizes of the two groups before they had settled, some-what higher figures than our estimate from other sources of information. They also indicate the death of 52% during the four years after first settling.

Physically, the Guayaki are an anomaly among Amerinds. They are fair, almost white-skinned, and short in stature relative to other Tupi-Guarani groups, and many males are prematurely bald, rare for Amer-ican Indians (figs. 2, 3) . They resemble closely people of Japanese or Korean origin. Cutis marmorata, a marble-like mottled or spotted discoloration, was observed on the shins of many of the children, apparently from long exposure to fires near which they sleep (fig. 4) . Characteristic tattoos, incised in fine parallel lines and raised by scarification, cover much of the bodies of most adults (fig. 5) . Seroepidemiologic data and historical information confirm

Fig. 2 A Guayaki man, probably in his 30's, demonstrating short stature, fair complexion, and unusual facial appearance of this group.

the existence of virgin soil epidemics after they had settled in their encampment and received frequent visitors (Brown and Gajdusek, '69b ); of the 88 listed as dead, 16 can be attributed to the measles epi-demic of July 1963, and 13 were relatively recent deaths of infants and young chil-dren. They all suffered, without exception, from continual coryza from overlapping upper respiratory infections during the first several years after settlement. Everyone had persistent purulent coryza and cough, which they claimed had been previously unknown to them.

The food gathering culture of the Gua-yaki centers about honey, insect larvae, and parts of the pindo palm (Cocos roman-zoffiana). They hunt small game with the largest bows and arrows to be found in the hemisphere. They are totally devoid of any organized agriculture or animal husbandry, or were so at the time of their emergence. While on hunting expeditions they made thatched-hut camps to last two or three days. Nudity is the norm. Guayaki crafts are confined chiefly to crude weaving, tool making and food preparation. Before settling, the Guayaki were extremely timid and fled when they encountered strangers.

More details about this expedition can be found in the work of Gajdusek ( '63) and a succinct summary of other data in Brown and Gajdusek ( '69b); historical an-notation in the work of Lozano (1873) , Mayntzhusen ( '25) and Bertoni ( '41) ; a summary of the film taken on this visit in the appendix to a monograph by Sorenson and Gajdusek ( '66) ; and the most reliable early sources of physical anthropologic in-formation in the works of La Hitte and ten Kate (1897) . Susnik ('61, '62), Cadogan and Colleville ('63, '64), Clastres and Sebag ( '63) , and Clastres ( '64) pro-vide recent ethnographic information.

Northwest Paraguay. To the north and west of the Paraguay River lies the Gran Chaco, more than 150,000 square miles of wastes, arid and sparsely populated, dry but briefly flooded during rainy seasons, densely vegetated with almost impene-trable brush. These wastes stretch north-ward to Brazil, melting southward into the Pampas of Argentina, and westward to the salt marshes and sub-Andean hills of south-eastern Bolivia. The unfriendly conditions

GENETIC STUDIES IN PARAGUAY 3 2 1

>i i > >

Fig. 3 Premature baldness occurs commonly among Guayaki men (estimated age, late 30's).

of the Chaco predisposed it to traversal, rather than colonization, by the Spanish. Despite early incursions by Jesuit and later Franciscan missions, the Chaco has re-

mained virtually unsettled until very re-cently. It is the last refuge of some of the most primitive hunter-gatherer groups of American Indians. Some 27 tribal groups

3 2 2 S. BROWN, GAJDUSEK, LEYSHON, STEINBERG, K. BROWN AND CURTAIN

Fig. 4 Cutis marmorata is found commonly on the shins of Guayaki Indians, especially of children. This is probably caused by the children's habit of sleeping on their side with their shins toward the fire.

still inhabit the area. Despite the apparent desolation, wild fruit , beans and edible bromeliads appear in abundance, as well as a variety of insects and small mam-

mals including deer, rhea and peccary, wild horses, monkeys and tapirs. Fish can be caught during the rainy season and shortly afterward.

GENETIC STUDIES IN PARAGUAY 3 2 3

Fig. 5 Ritual tattooing by scarification in fine parallel l ines results in this appearance on the back and arms among Guayaki adults.

Historically, little is known of the primi-tive Chaco Indians prior to the Conquest. The Guaicuruan, Matacoan and Lule-Vilelan groups, three of the seven primary

ethnic stocks, fought the Spanish in the early 16th Century and were largely mas-sacred. In spite of Jesuit and Franciscan attempts at missionization, the northern

3 2 4 S. BROWN, GAJDUSEK, LEYSHON, STEINBERG, K. BROWN AND CURTAIN

Chaco was never really settled: Argen-tinians occupied the southern part. Chaco Indian tribes remained culturally intact well into the 20th Century, long after most other Amerindian groups' cultures had dissolved. With the introduction of the horse, several tribes gave up agriculture and became savage warriors, often con-quering and subjugating other indigenes with hostility and fierceness equal to that with which they resisted the Spanish occu-pation. The War of the Triple Alliance, 1864-70, and the Chaco War of 1932-38 led to large scale loss of life among the Paraguayan mestizo and Indian popula-tions.

Culturally, representatives of six of the seven ethnolinguistic groups still survive in the Chaco. The Lule-Vilelan group is thought to be completely extinct. The cul-tures of Chaco Indian tribes resemble those of the North American Plains Indians. Of particular biological interest is the fact that several of the tribes practice intentional abortion and female infanticide. Mission-ary sources reveal a history of virgin soil epidemics with high mortality. Our own observations and seroepidemiological data (Brown and Gajdusek, '69a) indicate that Chaco tribes are currently completely sus-ceptible to certain common viruses. They have been swept by many virgin soil epi-demics. Seroepidemiology indicates such recent epidemics of measles, influenza, parainfluenza, and a number of picor-naviruses. Toxoplasma gondii seems highly endemic. The often reputed prevalence of syphilis is not supported by serological evi-dence. We observed cases of tuberculosis and bacterial pneumonia.

The colonization by German-speaking Mennonites, emigrants from Russia, some by way of Canada, has had a major effect upon Chaco Indian culture, second in im-portance to the influence of the missions. They have missionized and in some cases intermarried with the Indians, changed their way of dress, and taught them religious hymns. They, along with con-temporary missionaries, and mining or lumber camps, comprise the sole influence of Western culture on the Chaco Indians. The Mennonites are a pacifistic Protestant, highly inbred group, an offshoot of the Swiss Anabaptist movement from which

the Hutterites and Amish are also derived. They migrated to Holland, then Germany, and later Russia, and eventually to Canada to escape religious persecution. By the 1920's their Canadian colonists numbered more than 20,000. When certain rights were curtailed in Canada, a group of sev-eral hundred founded the first Chaco colony, Menno, in 1926. It was here the subjects were chosen for our blood group analysis. In the next twenty years two nearby colonies were founded for new emigrants f rom Poland and Russia, with more Canadians joining the original group. Two other Mennonite colonies were also founded in eastern Paraguay and three more in Uruguay, Argentina, and Colombia.

Mennonite marriage is strict and per-manent; average (not completed) family size is 4.8. Populations have been affected both,by World War II casualties among the males in the Neuland (Russian) colony, and generally, by the typhoid epidemic in 1926-27, which claimed more than 150 lives, about 10% of the Menno population. Fifty-one colonists from Menno served as subjects for our present blood group and serum genetic studies.

More complete general information con-cerning the Mennonites can be gathered from the works of Janzen ('44, '62), Fretz ( '53) , and Smith ( '50) ; and genetic stud-ies include the works of McKusick, Hostetler and Egeland ( '64a) ; McKusick, Hostetler, Egeland and Eldridge, ( '64b); and Jackson, Symon, Praden, Kaehr and Mann ( '68) . General background reading on Chaco Indian history and culture is to be found in the works of Brinton (1898), Kersten ( '05) , Lehmann-Nitsche ( '08), Metraux ( '46) , Belaieff ('46, '58), Beals ( '61) , and many citations in Steward ( ' 4 6 - 5 9 ) , and O'Leary ( '63). Table 1 lists all groups studied by ethnolinguistic group, site of collection of blood specimens and number of individuals studied. The eight Chaco tribes in the present study are dis-cussed below.

Toba. The Guaicuruan Toba number about 5000, and contain six subdivisions. Since the time of first European contact many have inhabited the region immedi-ately surrounding the Mennonite colony. The blood group genetic traits of 45 of the

GENETIC STUDIES IN PARAGUAY 3 2 5

approximately 120 now settled there are analyzed in this study.

Most Toba inhabit the region between the Pilcomayo and Bermejo Rivers. Many hundreds have been settled in two missions near the Pilcomayo. One group was settled in the Chiriguano mission at Machareti, subsequently adopting that language. An-other group lives in the region of Menno Colony at Laguna Pora.

Despite early baptism and missioniza-tion by Jesuits and Franciscans, the Toba were among the fiercest resistors of Chaco settlement, well into the present century. Intentional abortion and infanticide are commonplace in their culture, polygyny rare but present.

Detailed sources on the Toba include Metraux ('37, '46), Campana ( '03) , Koch-Grunberg ( '03) , Lehmann-Nitsche ( '23) , Karsten ( '23) , and Nordenskiold ( '13) .

Sanapana (Kyisapang). These 1000 Indians inhabiting the northwest area of the Chaco plains are of the Mascoian linguistic group. Their location at the time of first European contact is uncertain. The 105 in the present study were located at La Esperanza mission, about 16 km north-east of Menno Colony. They are hardly acculturated, practice little agriculture, and barely mix with the Toba and Lengua Indians of nearby Laguna Pora mission. Other Sanapana live in the area south of Puerto Sastre on the Salado and Galvan Rivers, and in the vicinity of Puerto Casado, all north and east of the Men-nonite colonies. They farm primitively, mainly melons and gourds. They were one of the few Chacoan groups to use the canoe as a means of travel. More complete material on this group can be found in the works of Metraux ( '46) , Koch-Grunberg ( '02) , and Loukotka ( '31) .

Lengua. The second of the Mascoian-derived groups studied is the Lengua, whose 10 or 11 bands total between 3000 and 4000 in number. Not to be confused with the Lengua-Enimaga of the Matacoan linguistic group, their habitat stretches westward from the Paraguay River to the Mennonite country. This is the same area inhabited by their forebears, the ancient Mascoi. Since 1887, many have lived in missions founded by English missionaries. The group in this study, learning to fa rm

at the Yalve Sanga Mission of the Men-nonites, numbers between 700 and 800, and shares the mission with 500 Chulupi. Among them, intentional abortion and female infanticide are commonplace. They have an apparent awareness of the trans-missibility of disease since they carefully burn the belongings of deceased persons, keep their sick, close relatives in isolation, and purify the air in the vicinity of death and burial with burning torches. Further information on this well-studied group can be found in the works of Hawtrey ( '01) , Grubb ( '14) , Coryn ( '22) , Koch-Grunberg ( '02) , and Metraux ( '46) .

Chulupi (Ashluslay). The Chulupi, comprised of four tribal groups whose total population is between 4000 and 5000, is derived from the Matacoan linguistic fam-ily whose tribes occupied a solid block across the Chaco from the Paraguay River to the Andes. The terms Chunipi and Tapiete, sometimes erroneously used as synonyms for the Chulupi, actually desig-nate other tribes. Chulupi presently occupy the southeastern Chaco in an area between the Mennonite colonies and the Pilcomayo River (southern border of Paraguay). They were not mentioned in any literature until the 19th Century and except for the studies of Nordenskiold, have received scant atten-tion. Early during the 20th Century they began to migrate southward to the Argen-tine sugar cane fields, where accultura-tion began, and they returned northward with European goods. During the Chaco War, many fled southward and relocated in Argentina. Although during the present century estimates of their population have ranged as high as 15,000, their numbers have dwindled recently. We found nearly 100 Chulupi at Marescal Estigarribia ( the former "Camacho") in a Catholic mission about 55 km northwest of Filadelfia, and a few, along with a group of Cheroti, in a settlement fur ther to the west. Nearly 100 were also in the Mennonite Mission at Filadelfia; others are known to be living at Yalve Sanga. Metraux ( '46) names four other Catholic Chulupi missions, admin-istered by the same order (Oblates of Mary Immaculate) , as the group at Marescal Estigarribia. The Chulupi live in the larg-est villages of any Chaco Indians, often as large as 1000. They were much displaced

3 2 6 S. BROWN, GAJDUSEK, LEYSHON, STEINBERG, K. BROWN A N D CURTAIN

by the Chaco wars, living in the areas most affected. They seized upon animal hus-bandry early, although they remained a non-equestrian tribe socially. They also have raised more crops than other groups. Further information about this group can be found in the writings of Palavecino ( '39) , Lehmann-Nitsche ( '36) , Norden-skiold ('13, '19), and Metraux ( '46) .

Cheroti (Munhui). The Cheroti, or Choroti, also of the Matacoan family, presently number about 2500 to 3000. At the time of first contact in 1736, they lived just north of the upper Pilcomayo River, near what is now the southwestern corner of Paraguay, less than 100 km downstream from their present day concentration. Earlier in the century they were noted to live close by the river. The two groups we encountered just west of Marescal Esti-garribia and at Santa Rosa, over 100 km west of there, were both 125 to 200 km away from the river. Santa Rosa is located near the military outpost, Fortin Teniente Hernandarias. The Cheroti have remained largely uninfluenced by the incursions of European and Chiriguano cultures. Their present culture is still that of a semi-nomadic desert band of honey- and food-gatherers who hunt and fish with bow and arrow. They appear extremely dirty and shy. Many are also able to speak the related Chulupi language, and bands of these two tribal groups seem to remain in contact with each other. Further facts about this group can be gathered from the writings of Rosen ( '06) , Metraux ( '46) , and from our own experiences with them, in the journal of Gajdusek ( '63) .

Ay ore (Moro). This violent tribe of honey- and salt-gatherers is of Zamu-coan origin, although debate concerning whether their forebears were Guaranocas or Morotocos is unsettled. Their total num-ber is approximately 3000, comprising at least 16 bands, whose names are known (15 more are thought to be extinct). Their first Jesuit contact (1691) and subsequent missionization provide evidence for early cross-contact with Indians of Chiquitoan and Guarani origin. Words of these deriva-tions are still found in the Ayore language. The lebensraum of this group stretches from the Mennonite colonies northwest-ward to Lake San Luis in east-central

Bolivia, encompassing a total area of more than 200,000 square miles, probably larger than any known human group not dependent upon the horse, camel or dog-sled for transportation. The mystery sur-rounding the relationship between the Paraguayan group ("Moro") and the Bolivian group ("Ayore," or "Ayoweo") be-came clear on our visit, when we learned that the Bolivian Protestant and Para-guayan Catholic and Mennonite missions shared common members who roamed freely back and forth. The Ayore roam in bands of 50 to 150, casually murdering intruders, a fact well documented by the history of their bloody contacts with at least three groups of religious missionaries who have successfully contacted them in the last 30 years. Settled groups of Ayore include those at the Tobite mission near Robore, 300 km east of Santa Cruz, Bolivia (New Tribes Mission); two missions (Zapoco and Bella Vista), about 25 km northeast of Santa Cruz, in the vicinity of San Ignacio; and the Catholic mission known as "Maria Ausiliadora" of the Salesian Order, which contains the group we visited 50 km north of Filadelfia, near Fortin Teniente Montania. This group, which recently numbered 200, has been reduced by a series of devastating epi-demics; these accounted for more than forty deaths during one epidemic alone. Seroepidemiologic data previously summar-ized (Brown and Gajdusek, '69a), and to be described in more detail in a fu ture pub-lication, lend support to this history. The demographic composition of the Ayore at the Salesian mission camp was as follows.

Population Mean age

M F T M F T

Study group (a l l ages ) 38 35 73 24 25 25

Adults * 28 28 56 30 29 30 Children * 10 7 17 9 8 9 * Ages of subjects were estimated, and adults in-

cluded all — 13 years, children all < 13 years.

Culturally, the Ayore are hunter-gather-ers, living in a shared pool of dirt and salivary and fecal microbes. They too prac-tice intentional abortion and female in-fanticide. Although they have not been intensively studied in anthropological lit-erature, the missions with whom they have settled have collected a wealth of informa-

GENETIC STUDIES IN PARAGUAY 3 2 7

tion about them, the best of which (Pen-cille and Pencille, '63) is unpublished. For further sources see the works of Kelm ('60, '63), Pencille ('60, '61a, '61b, '61c, '67), Pencille and Pencille ('60, '63), Janzen ( '62) , Norwood ( '62) , South America Indian Mission (n .d . ) , Susnik ( '63) , and our own journal and cinema (Gajdusek, '63, Sorenson and Gajdusek, '66) .

Guarayu This tribe, whose culture and origin are distinct from most other Chaco indigenes, is of the Tupi-Guarani family, sometimes assigned to the Chiriguano subdivision. Certain Guarani migrated westward in waves, starting in the late fifteenth century, from eastern Paraguay to the edges of the Inca Empire. The Chiriguano, Guarayu, and other groups settled in the Bolivian Cordillera, where their culture became successively modified by contact with the neighboring Chiquito and Mojo. They were missionized by Jesuits in the late seventeenth century, and by the Franciscans f rom 1840 to the present. In the period following Bolivian independence, many Guarayu left their missions, returning to the bush. Their sub-sistence culture today is largely aboriginal: they hunt with bow and arrow, and use poisons for both hunting and fishing, and practice collectively primitive agriculture, primarily involving maize and manioc. The earlier, large Guarani communal house has been replaced by a one family dwelling. The preferred marriage for a female was with maternal uncle or cross-cousin. Their primary Guarani culture included ceremo-nial cannibalism, nudity and communal-ism, all of which are now abandoned. Pres-ent population is about 5000, in mission locations documented by Metraux ('48). At least 700 died in the influenza pandemic of 1917-18. Detailed information on this group is found in Metraux ('29, '48) , D'Orbigny (1839, vol. 1), Nordenskiold ( '17) , Pierini ( '10) , Schmidt ( '36) , and Snethlage ( '35) .

Tapiete. This group consists of about 2000, in five bands. They have adopted the Guarani language of their Chiriguano neighbors to the west, with whom they have had a servant relationship for cen-turies. Despite the many influences on them by Chiriguano, their material cul-

ture remains essentially similar to Mata-coan groups, such as the Chulupi (who are often misnamed "Tapiete"). The Tapiete have been misnamed "Yanaiga," under which title they are known to have been at war with the Siriono for many years. They have also waged war with the Tsirakua, near Izozog, where several bands have settled since the Chaco War, and only recently made peace with the Caraja. Their original area of habitation was along the north bank of the upper Pil-comayo, perhaps pushed southward by white settlers. Since the Chaco War, they settled near Fort Oruro, Izozog, and at Marescal Estigarribia. The band included in this study, called "Guasurangue" after their old chief (Colleville, '72), is settled at the latter camp. Further information about this group is found in Metraux ('46, '49), Belaieff ( '46) , and citations noted in Steward ( '46- '59) , Stewart ( '52) , and Key and Key ( '61) .

MATERIALS AND METHODS

Specimens. The 591 blood specimens were collected between September 3 and September 22, 1963 from the groups shown in table 1. Blood was collected in sterile vacuumatic venules, was stored at 0°C within 24 hours and was then shipped to the National Institutes of Health via air freight from Asuncion. When it arrived, separation of clots from serum, and dis-tribution of serum aliquots was performed. In almost all cases this took place within five days of the time of collection. At that time clots were stored at 4°C and serum at — 20 °C until used for genetic or im-munologic tests.

Red cell genetic studies. Immediately upon storage at 4°C the clots were studied for red cell groups and types. The red cells were typed for the following antigens: A, A,, B, M, N, S, s, U, He, P., C, Cw, D, E, c, e, K, Kp\ Kp\ k, Le', Fy', and Di\ The methods for storing clots, testing and computing results are similar to those de-scribed by Pollitzer, Hartmann, Moore, Rosenfield, Smith, Hakim, Schmidt, and Leyshon ( '62) , and Pollitzer, Waggoner and Leyshon ( '67) . The method of prep-aration of hemoglobin solution and the criteria for defining thalassemia trait are described by Curtain, Kidson, Gajdusek,

3 2 8 S. BROWN, GAJDUSEK, LEYSHON, STEINBERG, K. BROWN A N D CURTAIN

TABLE 1 Ethnic origin of specimens studied

Ethnic group studied Site of collection Number bled

Southeast Paraguay Tupi-Guarani

Guayaki Ache Kwera Ache Gatu

Total Guayaki

Northwest Paraguay Guaicuruan

Toba

Mascoian Sanapana

Lengua

Total Lengua

Matacoan Chulupi (Ash lus lay )

Total Chulupi

Cheroti ( M u n h u i )

Total Cheroti

Z amucoan Ayore (Moro)

Total Ayore

Tupi-Guarani Guarayu Tapiete

Intertribal Mixtures Lengua-Maca Lengua-Chulupi Chulupi-Cheroti Chulupi-Guarayu

Total Mixtures

Mennonite Colonists Total in study

Arroyo Moroti Arroyo Moroti

Loma Plata

La Esperanza Yalve Sanga Menno Colony, Loma Plata

Marescal Estigarribia Filadelfia Settlement west of Marescal

Estigarribia

Santa Rosa Settlement west of Marescal

Estigarribia

Salesian Camp Menno Colony, Loma Plata

Marescal Estigarribia Marescal Estigarribia

Yalve Sanga Yalve Sanga Filadelfia Marescal Estigarribia

Menno Colony, Loma Plata

40 22

45

104 47

3

27 91

21

10

72 1

15 19

51

62

45

104

50

125

31

73

15 19

16

51 591

and Gorman ( '62) , and the method of hemoglobin electrophoresis employed is also described by Curtain ( '62). The test for identification of G6PD is that of Rom-berg and Horecker ( '55) .

Serum, genetic studies. Serum gamma Globulin Gm and Inv factors were tested according to the method described else-where (Steinberg, '62), the only modifica-tion being the change in terminology from the lettered to the numbered Gm factors (WHO, '65). (The samples were tested prior to 1965 for G m ( l ) , Gm(2) , Gm(5 ) ,

Gm(6) , and I n v ( l ) . Except for Inv(2) , these were the only allotypes for which sera were then available. Serum hapto-globin testing for all tribal groups was per-formed by Dr. Kenneth Brown, using a discontinuous acrylamide gel technique described in the E.C. Apparatus Technical Bulletin 134, 1968 (Brown and Johnson, 7 0 ) .

RESULTS

Phenotypic incidence and gene fre-quencies for blood groups and serum

GENETIC STUDIES IN PARAGUAY 3 2 9

genetic traits are given in tables 2 to 8. Results based on testing with 19 antisera for nine major blood groups cited above, for G6PD deficiency, haptoglobins and Gm and Inv immunoglobin markers are de-scribed. Gene frequencies have been cal-culated for all groups containing ten or more individuals bled and are given with two significant digits. MNSs results were estimated by gene counting methods de-scribed by Mourant ( '54) and modified by one of us (SMB) since maximum likeli-hood estimates are not appropriate when dealing with small population isolates not in Hardy-Weinberg equilibrium. ABO gene frequencies have been calculated using the method of Bernstein ( '30) where indicated. Simple gene counting for Rh frequencies started with the assumptions that r = O for all Indian groups, and R° = O for Men-nonites. These assumptions were based upon both empirical expectations and maximum likelihood estimates using the computer program of the Human Genetics Branch, National Institute of Dental Re-search, based on the method of Kurczynski and Steinberg ( '67) . Gene counting was also the method used in cases where Kell, or serum factors required computation (i.e., were not homogeneously positive or negative for all members of the group in question). For all other cases requiring computation, for the other blood groups, results were estimated by the simple bi-nomial square in which the phenotypic incidence of the recessive trait is assumed to be the square of its gene frequency. For the Duffy group, Fy" testing was not done. The assumption we made eliminates the existence of the third (amorphic) gene, Fy, found in African populations (see Chown, Lewis and Kaita, '65), assumed to be present when Fy° and Fy" are both absent. We have assumed Fyb to be a clas-sical recessive gene whose frequency is the square root of the proportion 1 minus the occurrence by Fy" positive. Since Le(b) and H testing were not performed Le and le frequencies could not be estimated.

Blood group genetic results ABO results. ABO tests yielded homo-

geneous results, except for the Mennonites, and are presented in table 2. All Amer-indians tested were Group O, except for

one individual described as Chulupi-Guarayu mixture, who was group B.

The ABO results for the Mennonites are somewhat unusual for Caucasians of west-tern European origin. Their total incidence of phenotypic Group A was but 37% , and the significance of this finding will be dis-cussed below. Their incidence of Group O was unusually high and none of them showed the presence of Group B.

MNSs-U-He results. The results for this group are presented in table 3. The high-est frequencies were found for MMSs, MMss, MNSs, and MNss. The unusually high incidences of the last mentioned phenotype in the Guayaki and Tapiete are worthy of note. The Ay ore showed a re-markably high phenotypic incidence of both M and S, higher than 90% for each.

For all Indian tribes tested, of the four possible chromosome combinations, the highest frequencies were for chromosome Ms, except for the tribes of the Mascoian ethnolinguistic group and for the Ayore, where MS predominated. These MNSs re-sults are discussed at greater length below, where comparisons with other indigenous groups were made. All persons tested were Henshaw (He) negative and U positive. The individuals tested for U and He were identical to those tested for Duffy (Fy*), numbers noted in table 6, except that only 7 of the 8 of mixed tribal derivation were tested for U and He.

Pi results. The phenotypic frequencies of Pi positive individuals among the Indians tested was quite high, only two groups being lower than 50%. The Ayore were the lowest, with a 32% positive result. The Guayaki were homogeneously positive. Gene frequencies ranged from 0.18 for the Ayore to 1.0 for the Guayaki,

Rh-Hr results. Classically, Amerin-dians have complete absence of chromo-some r (cde) and thus of Rh-negative in-dividuals. This result bore true in our results, as shown in table 4. There it can be seen that among all Indians bled, there was no instance of a homogeneously Rh-negative r / r (cde/cde) individual; further-more, there were only very few instances of significant numbers of individuals occurring in the two columns where r (cde) was an alternative to R° (cDe) for one of a pair of chromosomes. The high-

3 3 0 S. BROWN, GAJDUSEK, LEYSHON, STEINBERG, K. BROWN A N D CURTAIN

TABLE 2 ABO blood group. Distribution of phenotypes and gene frequencies

Ethnic group studied Number

Phenotypic results Gene frequencies Ethnic group studied

Number O A B AB O A B

Southeast Paraguay Guayaki

Ache Kwera 40 40 0 0 0 1.00 0.00 0.00 Ache Gatu 22 22 0 0 0 1.00 0.00 0.00

Total Guayaki 62 62 0 0 0 1.00 0.00 0.00

Northwest Paraguay Toba 14 14 0 0 0 1.00 0 .00 0.00 Sanapana 104 104 0 0 0 1.00 0.00 0.00 Lengua 50 50 0 0 0 1.00 0.00 0.00 Chulupi 86 86 0 0 0 1.00 0 .00 0 .00 Cheroti 31 31 0 0 0 1.00 0.00 0 .00 Ayore (Moro) 73 73 0 0 0 1.00 0.00 0.00 Guarayu 15 15 0 0 0 1.00 0.00 0 .00 Tapiete 19 19 0 0 0 1.00 0 .00 0.00 Intertribal Mixtures 12 11 0 1 0 0.96 0.00 0 .04 Mennonite Colonists 51 32 19 1 0 0 0.79 0.21 0 .00

• Includes 15 type Ai and 4 type A2.

est occurrences of such incidences were the Guarayu, the Toba and the Guayaki. The maximum likelihood estimation of gene frequencies, however, showed the probable frequency of r (cde ) to be O in all cases, favoring small, finite frequencies for R° (cDe).

It is noted that R' (CDe) and R2 (cDE) are the most frequently present chromo-somes. The incidences of R2 (cDE ) tended to be low for groups where R' (CDe) was high, and conversely.

R' (CDE) was present in all but the Guayaki, Guarayu, and Mennonites (0 .01-0.25), the incidence amongst the Toba, Sanapana, Lengua, and Tapiete being ex-traordinarily high (table 4) .

The Mennonites showed high phenotypic frequencies for Rh.Rh, (CCDee), RhiRh* (CcDEe), and Rh.rh (CdDee). Their chro-mosome frequencies are not unusual for Caucasians and will be discussed below. All individuals were tested for rh" (Cw), except the 62 Guayaki, and all were nega-tive.

Kell-Cellano results. All of the Indians tested were homogeneously positive for k and Kp\ and negative for K and Kp*, as shown in table 5. In the Mennonites all persons were positive for k and Kp", but also three were positive for Kp* and one for K. It should be noted that while gene frequencies were estimated as if for two separate loci, each with two alleles, this

group is probably one complex locus of the cistron type similar to MNSs, and chromo-some frequencies could be computed for K-Kp', K-Kp\ k-Kp• and kKpb.

Lewis group results. Since results are based on testing only for the Le antigen, no gene frequencies have been estimated.

Several groups showed complete ab-sence of Le": the Mennonites, Guarayu, Tapiete, Ayore, Chulupi, Sanapana, and the Ache Kwera band of the Guayaki.

Duffy results. All groups tested showed a particularly high phenotypic frequency of Fy\ Of the three Tupi-Guarani groups tested, two (the Guayaki and the Tapiete) were all positive and the third, the Gua-rayu, had only one negative. The Mennon-its were 71% positive. Results are sum-marized in table 6.

Diego results. The Ayore, Guarayu, Toba and Tapiete were uniformly negative for the Diego factor, and the Chulupi had only one positive (table 6) . All other Indian groups' frequencies for Di" were low but positive.

G6PD and Hemoglobin results. G6PD deficiency and thalassemia trait were absent for all Indians and Mennonites studied.

Serum genetic results Serum globulin results. Results for the

Gm and Inv serum markers and hapto-globins are given in tables 7 and 8. Only

TABLE 3 MNSs blood group. Distribution of phenotypes and chromosome frequencies

Phenotypic results

Ethnic group studied M MN N

jutheast Paraguay Guayaki

Ache Kwera

Ache Gatu

Total Guayaki

Number

SS Ss ss SS Ss Sf SS Ss s: 5 Chromosome frequencies

Number No. % No. % No. % No. % No. % No. % No. % No. % No. % MS Ms NS Ns

40 0 0 4 10 9 22 0 0 3 8 17 42 0 0 0 0 7 18 0.09 0.49 0 . 0 0 0.42

22 1 5 • 6 2 7 14 64 0 0 1 5 0 0 0 0 0 0 0 0 0.20 0.77 0 . 0 0 0.02

62 1 2 10 16 23 37 0 0 4 6 17 27 0 0 0 0 7 11 0.13 0.59 0 . 0 0 0.28

orthwest Paraguay

Toba 10 3 30 3 30 0 0 1 10 2 20 1 10 0 0 0 0 0 0 0.65 0.15 0.00 0.20

Sanapana 89 25 28 36 40 11 12 3 3 7 8 5 6 0 0 2 2 0 0 0.52 0.38 0.05 0 .06

Lengua 39 9 23 6 15 5 13 1 3 6 15 4 10 1 3 4 10 3 8 0.37 0.29 0.12 0.22

Chulupi 66 5 8 21 3 2 19 29 1 2 8 12 10 15 0 0 1 2 1 2 0.25 0.58 0.04 0 .13

Cheroti 28 3 11 4 14 6 21 0 0 8 29 4 14 0 0 1 4 2 7 0.28 0.40 0.06 0 .26

Ayore (Moro) 72 36 50 27 38 4 6 0 0 5 7 0 0 0 0 0 0 0 0 0.72 0.24 0.00 0.04

Guarayu 13 0 0 4 31 2 15 2 15 0 0 3 23 0 0 1 8 1 8 0.25 0.41 0.10 0.25

Tapiete 19 0 0 3 16 3 16 1 5 1 5 10 53 0 0 0 0 1 5 0.16 0.47 0.00 0.37

Intertribal Mixtures 8 2 25 0 0 0 0 1 12 3 38 2 25 0 0 0 0 0 0 — — — ' —

Mennonite Colonists 51 3 6 4 8 7 14 2 4 4 8 25 49- 0 0 1 2 5 10 0.16 0.42 0.03 0.39

All U Positive. All He Negative.

TABLE 4 Rh-Hr blood group. Distribution of phenotypes and chromosome frequencies

Phenotypic results Chromosome frequencies

Ethnic group studied Rh,Rh, Rh,Rh2 Rh2Rh2 RhiRh, RhjRh, Rh,Rh, Rh,Rh„ i Rh2Rh„» rh (CCDee) (CcDEe) (ccDEE) (CCDEe) (CcDEE) (CCDEE) (CcDee) (ccDEe) (ccddee) Ri R2 Ro R, T

Number No. % No. % No. % No. % No. % No. % No. % No. % No. % (CDe) (cDE) (cDe) (CDE) (cde)

>utheast Paraguay

Guayaki Ache Kwera 40 10 25 14 35 5 12 0 0 0 0 0 0 6 15 5 12 0 0 0.50 0.36 0.14 0.00 0.00

Ache Gatu 22 14 64 6 27 0 0 0 0 0 0 0 0 1 5 1 5 0 0 0.80 0.16 0.05 0.00 0.00

Total Guayaki 62 24 39 20 32 5 8 0 0 0 0 0 0 7 11 6 10 0 0 0.61 0.29 0.11 0.00 0.00

orthwest Paraguay

Toba 10 1 10 2 20 2 2 0 2 20 0 0 1 10 1 10 1 10 0 0 0.35 0.35 0.10 0.20 0.00

Sanapana 97 5 5 6 6 53 55 2 2 20 21 11 11 0 0 0 0 0 0 0.09 0.68 0.00 0.23 0.00

Lengua 46 6 13 12 26 6 13 11 24 4 9 4 9 1 2 2 4 0 0 0.39 0.33 0.03 0.25 0.00

Chulupi 70 12 17 32 4 6 20 2 9 2 3 2 3 0 0 2 3 0 0 0 0 0.43 0.53 0.01 0 .03 0.00

Cheroti 29 5 17 13 45 3 10 2 7 6 21 0 0 0 0 0 0 0 0 0 .43 0.43 0.00 0.14 0.00

Ayore (Moro) 73 39 53 28 38 4 5 0 0 2 3 0 0 0 0 0 0 0 0 0 .73 0.26 0.00 0.01 0.00

Guarayu 13 4 31 4 31 3 2 3 0 0 0 0 0 0 1 8 1 8 0 0 0.50 0.42 0.08 0.00 0.00

Tapiete 19 0 0 0 0 10 5 3 5 26 3 16 1 5 0 0 0 0 0 0 0.13 0.61 0.00 0.26 0.00

Intertribal Mixtures 10 0 0 2 20 8 8 0 0 0 0 0 0 0 _ 0 0 0 0 0 0 0.10 0.90 0.00 0.00 0.00

Mennonite Colonists 51 15 29 18 35 0 0 0 0 0 0 0 0 12 24 1 2 5 10 0.59 0.19 0.00 0.00 0 .23

All individuals except the 62 Guayaki Indians were tested for Cw and all were Cw negative. ' The phenotype CcDee is assumed to be Rh|Rh„ for Indians, Rhirh for Mennonites. 2 The phenotype ccDEe is assumed to be Rh2RhQ for Indians, Rh2rh for Mennonites.

GENETIC STUDIES IN PARAGUAY 3 3 3

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G m ( l ) and Gm(2) were found among the Indian samples. The frequency of Gm' was estimated by computing the square root of the homozygous case; of Gm' by gene counting; of Gm',z by summing the two and subtracting from 1.0. The frequency of Inv' was also computed by the square root method, its dominance over its absence being assumed on the basis of much pub-lished data. Haptoglobin frequencies for Hp 1 and Hp 2 were computed by the simple gene counting method, after ex-cluding from totals all specimens which tested type O-O. (Although this is some times a genetically determined trait, it is also environmentally determined in some cases because of hepatic disease or other acquired pathologic conditions).

The Gm and Inv results are not in dis-agreement with other results for South American Indians (Salzano and Steinberg, '65; Shreffler and Steinberg, '67; Salzano, Steinberg and Tepfenhart, '73). In the literature putatively pure South American Indian populations lack the allotypes Gm(5) and Gm(6) , and they show a pre-dominance of the phenotype G m ( l ) over the phenotype G m ( l , 2 ) and, therefore, a relatively greater frequency of Gm' com-pared to Gm' *. In the paper cited, which deals with Indians from various regions of Brazil, frequencies for Gm' range from 0.55-0.89. There is a negligible incidence ( < 0 . 0 1 - 0 . 0 4 ) of the haplotype Gm which did not occur among any of the Indian groups tested in our present study. Our results are interesting in that the Guayaki tribe, specifically the Ache Kwera band, show a greater frequency of Gm(l,2) than G m ( l ) . The Tapiete tribe, also Tupi-Guarani, had a 100% phenotypic fre-quency for G m ( l ) , but only 19 samples were tested. In general, the gene fre-quencies form Gm' were high, above 0.8 for all tribal groups except the Guayaki. It is of interest that two of the three Tupi-Guarani groups which were included in our sample also were highest for Gm' fre-quency. The occurrence of the unusually low Gm' frequency for Ache Kwera group (Guayaki), also Guarani-derived group, is only one of the myriad of anthropological phenomena which set this tribe apart from others. As expected, the Mennonites were

w CO

TABLE 6 Other blood group antigens studied: Results for P, Duffy, Diego, and Lewis antigens. Distribution of phenotypes and gene frequencies

Ethnic group studied

Phenotypic results

Number Pi

No. % No. Le»

No. Fy«

No. Di«

Gene frequencies

Fy Dia

a v o

z o >

a d CO W «

5 CO X O z CO H W W w V o

w SB o ^ z > z a o c » H >

Southwest Paraguay Guayaki

Ache Kwera Ache Gatu

Total Guayaki

Northwest Paraguay Toba Sanapana

Lengua

Chulupi

Cheroti Ayore ( Moro )

Guarayu Tapiete

Intertribal Mixtures

Mennonite Colonists

4 0 22 62

10 87 8 9 96 39 4 5 65 66 28 72 73 12 19

7 8 9

51

4 0 22 62

4

86

18

4 2 24 23

5 16

4

41

100 100 100

4 0

97

4 6

64 86 32

4 2 84 57

80

0 5 2

10 0

8

0

11 0

0 0 0

40 22 62

7 78

38

61

25 68

11

19

8

36

100 100 100

70 90

97

94

89 94

92 100

100

" 71

5 6

11

28

0

I 4

0 0 0

12 27 18

29

0

2 14

0 0 0

11 0

1.00 1.00 1.00

0.23

0.82

0.27

0 .40

0.62 0.18

0.24 0.60

0.56

1.00 1.00 1.00

0.45 0.68

0.87

0.75

0.67 0.76

0.71 1.00

0.46

0.06 0.15 0.09

0.00

0.16

0.00

0.01 0.07

0.00 0.00 0.00

0.00

TABLE 7 Gm and Inv serum immunoglobulin system. Distribution of phenotypes, Gm Haplotype frequencies, and Inv1 allele frequency '

Phenotypic results

Gm Haplotype or allele frequencies

Ethnic group studied Number No. %

1,2 No. %

5 1,5 1 ,2 ,5 No. % No. % No. % No.

Inv Inv a-f

Gm

1, 2

Inv

(1) (-1)

O M z M H l-H O CO H G a »—« M (O

> > o a >

Southwest Paraguay Guayaki

Ache Kwera 4 0 15 38 2 5 6 2 0 0 0 0 0 0 9 2 2 0.61 0.39 0.00 0.12 0.88 Ache Gatu 21 14 67 7 3 3 0 0 0 0 0 0 7 33 0.82 0.18 0.00 0.18 0.82

Total Guayaki 61 29 48 3 2 52 0 0 0 0 0 0 16 26 0 .69 0.31 0.00 0.14 0.86

Northwest Paraguay Toba 42 34 81 8 19 0 0 0 0 0 0 39 93 0.90 0.10 0.00 0.73 0.27 Sanapana 97 69 71 28 2 9 0 0 0 0 0 0 81 84 0.84 0.16 0.00 0.59 0.41 Lengua 45 41 91 4 9 0 0 0 0 0 0 39 87 0 .95 0.05 0.00 0.63 0.37 Chulupi 121 85 70 3 6 30 0 0 0 0 0 0 4 3 3 6 0.84 0.16 0.00 0.20 0.80 Cheroti 28 24 86 4 14 0 0 0 0 0 0 5 18 0 .93 0 .07 0.00 0.09 0.91 Ayore (Moro) 71 70 99 1 1 0 0 0 0 0 0 4 2 59 0.99 0.01 0.00 0.36 0.64 Guar ay u 14 13 93 1 7 0 0 0 0 0 0 11 79 0 .96 0.04 0.00 0.54 0.46 Tapiete 19 19 100 0 0 0 0 0 0 0 0 17 89 1.00 0.00 0.00 0.68 0.32 Intertribal Mixtures 15 15 100 0 0 0 0 0 0 0 0 8 53 1.00 0.00 0.00 0.32 0.68 Mennonite Colonists 51 0 0 0 0 49 96 1 2 1 2 17 33 0.01 0.01 0.98 0.18 0.82

> All samples were tested for Gm (1, 2, 5, 6 ) and Inv (1) . Only positive reactions indicated.

W W cn

3 3 6 S. BROWN, GAJDUSEK, LEYSHON, STEINBERG, K. BROWN A N D CURTAIN

TABLE 8 Serum haptoglobins. Distribution of phenotypes and gene frequencies

Phenotypic results

Ethnic group studied Number No. No.

2 - 1 No.

2 - 2 No.

0-0 Gene frequencies

Hpl Hp2

Southeast Paraguay Guayaki

Ache Kwera 40 0 0 12 30 27 68 1 2 0.15 0.85 Ache Gatu 22 3 14 9 41 10 45 0 0 0.34 0 .66

Total Guayaki 62 3 5 21 34 37 60 1 2 0 .22 0.78 Northwest Paraguay

Toba 45 5 11 26 58 14 31 0 0 0 .40 0 .60 Sanapana 101 56 55 39 39 6 6 0 0 0.75 0.25 Lengua 48 13 27 28 58 7 15 0 0 0.56 0.44 Chulupi 125 37 30 58 46 29 23 1 1 0.53 0.47 Cheroti 40 15 38 18 45 5 12 2 5 0 .63 0.37 Ayore (Moro) 72 12 17 35 49 25 35 0 0 0.4 1 0.59 Guarayu 15 5 33 5 33 2 13 3 20 0.62 0.38 Tapiete 19 1 5 12 63 6 32 0 0 0 .37 0.63 Intertribal Mixtures 16 2 12 9 56 5 31 0 0 0.41 0.59 Mennonite Colonists 47 5 11 27 57 15 32 0 0 0.39 0.61

homogeneously high for Gm3, a haplotype characteristic of Caucasoids.

The frequency of Inv' ranged from 0.09 for the Cheroti to 0.73 for the Toba. The Guayaki figure was only just higher than the Cheroti, in that respect. These two rep-resent the lowest Inv' frequencies thus far reported from South America, as reviewed by Salzano et al. ( '73) . The phenotypic and haplotype frequencies are shown in table 7.

Haptoglobin results. Haptoglobin re-sults are summarized in table 8. The occur-rence of phenotype 1-1 and the gene fre-quencies for Hp 1 were lower than is usual for American Indians. In the case of the Guayaki, this was one of the lowest Hp 1 gene frequencies ever recorded. This was in part due to the complete absence of type 1-1 among 39 members of the Ache Kwera tribal band tested, although 31% of the Ache Kwera were heterozygous. Gene frequencies of Hp 1 ranged from 0.15 for the Ache Kwera (Guayaki) to 0.75 for the Sanapana.

DISCUSSION

ABO. These homogeneous data for Group O are consistent with results in studies of other South American Indians (Salzano, '57). It is also consistent with the results which have been reported for Paraguayan or Chaco Indians (Mazza and Franke, '27a, '27b; Vellard, '34; Ribeiro, '34; Ribeiro, Berardinelli and Roiter, '35;

Alvarez, '39; Urizar, '42; Salzano, '57; Smith, '50; Matson, Sutton, Swanson and Robinson, ' 68a, '69), which include an early study (Vellard, '34), in which three Guayaki Indians were bled. Among North American Indians relatively high fre-quencies of A do occur, as annotated by the early discoveries among the Blood and Blackfoot, by Matson and Schrader ( '33) , a phenomenon which continues to mani-fest itself in the northwest U. S. and Canada (e.g., the recent study of B. M. Alfred et al., '70, of the Anaham) . It is consistent with the existence of a potenti-ally lethal disease, hemolytic disease of the newborn, caused by ABO incompatibility, that given a population which has a high frequency for O, that frequency should move toward 1.00 with time.

We found an unusually low incidence of Group A in the Mennonite group. There are at least three other studies of blood groups for isolates derived from Dutch or Swiss Anabaptists (McKusick, Bias, Norum and Cross, '67; Steinberg, Bleibtreu, Kur-czynski, Martin, and Kurczynski, '67; and Jackson, Hymon, Pruden, Kaehr and Mann, ' 6 8 ) .

The Paraguayan Mennonites are differ-ent from their North American relatives. While the incidences of blood Group A among North American Amish are 54% and 74% in the two demes, Holmes County, Ohio, and Lancaster County, Penn-sylvania, that of our Mennonite population

GENETIC STUDIES IN PARAGUAY 3 3 7

was 37%. The chief difference appeared to be a decreased incidence of Group Ai, in which the Lancaster County Amish incidence is 65% and the Mennonites 29% . The A2 incidences were 9% and 7% , respectively, for Amish and Mennonites. Since the Mennonites we studied have a much higher incidence of Group O and a lower incidence of Group A than we might have expected, it should be pointed out that while the Amish are thought to be derived from Berne Swiss, the Mennonites affirm their origins to be Dutch (Smith, '50). This becomes significant in that the Alpine Swiss are one of two groups in Europe known to be peculiarly high in their incidence of Group A (the other being Scandinavia).

Steinberg et al. ( '67) studied the Hut-terites, an Alpine Swiss Anabaptist group who settled in the northcentral U. S. and central Canada in 1874-79. In that study extensive blood group data are provided in the form of gene frequencies for one sub-isolate (group of colonies) of the three now existent. The gene frequency for Group A (0.325) is still considerably higher than our Mennonites' figure of 0.21, and their A2 frequency considerably higher. The Hutterites' frequency for gene A (0.325) is quite similar to that given by Mourant for the Berne-derived Swiss (0.303), as is their frequency for O (0.653; Berne Swiss 0.629), and B (0.022; Berne Swiss: 0.067). Further data on 1,386 Amish in Adams County, Indiana, were recently published by Jackson et al. ( '68) . Their phenotypic incidences of Groups A ( 6 4 . 6 % ) and O ( 1 5 . 6 % ) show them to be similar to the other two Amish demes, with phenotypic frequencies of A quite high, O quite low, and lying between the Holmes and Lancaster County figures. Thus, the Mennonites' high frequency for O (0.79) and low frequency for A (0.21) contrasts markedly with the data provided from North American Amish and Hutterite demes.

These data probably illustrate the effect of genetic drift in well defined isolates thought to be of similar ethnic origin in a period of less than two centuries. However, neither selection nor differences in origin can be ruled out as causes. If drift is re-sponsible, it is interesting that the Amish

and Hutterites all illustrated an upward drift in the frequency of A, downward of O, while the case with Paraguayan Men-nonites was just the reverse.

MNSsU and Henshaw. When analyzed for gene frequencies of M, N, S, and s individually, the general rule is a low fre-quency of N and a low frequency for S among American Indians. In that respect frequencies for N are quite high among the Paraguayan Indians, being above 0.30 in several cases (Ache Kwera band of Guayaki, Lengua, Cheroti, Guarayu, and Tapiete). In one case (Ache Kwera) the frequency was greater than 0.4. Frequen-cies for N at or near O have been reported, particularly in Peru (Matson, Sutton, Swanson and Robinson, '66a), but the ex-tremely low frequency among the Ayore is notable in that respect. The high s and low S rule was observed in all groups ex-cept the Sanapana, Toba, and Ayore (all with s < 0.5); the Ayore case of s = 0.28 was the lowest; the only two cafees of South American Indians with lower frequencies for s are the Isconahua of Peru and the Chacobo of Bolivia (Matson et al., '66a, '66b), in both of which Ms was the pre-dominant form of s present.

When analyzed for frequencies of the individual chromosomes, one expects that Ms be present in the highest frequencies, and NS in the lowest. In general, this was observed for the Paraguayan groups; however, in four cases the NS chromosome was totally absent: both Guayaki bands, the Ayore and the Tapiete. It should be noted that all of these but the Ayore are of the Tupi-Guarani ethnolinguistic group. Similar results have been observed for the Zamucoan-speaking Moro (Ayore) and Chamococo (Matson et al., '68a). The predicted serial order of frequencies, Ms > Ms > Ns > NS did not occur in just those groups where MS was higher than Ms.

All individuals tested were positive for the U antigen, including the 51 Mennon-ites. Significant percentages of U-negative individuals occur in African populations.

The Mennonites' results for MNSsU were not dissimilar to expected values for a western European group, with a nearly absent NS, and high values for Ns and Ms. Ms was higher than would be usual for

3 3 8 S. BROWN, GAJDUSEK, LEYSHON, STEINBERG, K. BROWN AND CURTAIN

such a group, for it is uncommon that Ms be higher than 0.30 for groups other than those of Celtic (Welsh, Scotch, or Irish) origin (Mourant, '54). Steinberg et al. ('67) did show that MNSs gene frequencies exhibit a remarkable amount of drift in their study of Hutterite colonies. Data from the Jackson et al. ( '68) study show that our Mennonite results are much closer to phenotypic MNS frequencies among the Swiss than among the Indiana Amish. All individuals were negative for Henshaw antigen (He) , which is found in low frequencies among African groups, absent in Caucasians.

Pi. Frequencies for P, are generally high in Ameridians, but vary greatly. The homogeneity illustrated by the Guayaki (both bands) in their total positivity for Pi-factor has been seen in few other tribes of American Indians: e.g., the Chacobo, the Isconahua, and the Yagua (Matson et al., '66b) and Calchaqui (Matson et al., '69). The frequency for P, of 0.56 for the Mennonites is similar to the European fre-quencies, which run above 0.50. The P, gene frequency for the Lancaster County Amish was 0.573 (McKusick et al., '67).

RH. The most unusual finding among the Rh results were astoundingly high fre-quencies of the chromosome R" (CDE). Paraguayan Indians tend to show a rela-tively high chromosome frequency for R' (CDE); our data, in that respect, are not unlike that for the only other Rh study of Paraguayan Chaco Indians (Matson et al., '68a). The Toba, Sanapana, Lengua and Tapiete frequencies of R' (CDE) are among the highest ever found in any human group; the highest known value for R' (CDE) outside South America, to our knowledge, is that among Kurdistani Jews (0.09) (Gurevitch and Margolis, '63); the highest reported for Amerin-dians to date is 0.275 (Matson et al., '68a). We do not believe that frequencies higher than 0.1 have been reported for any group other than five Paraguayan groups: the Sanapana, Lengua, Cheroti, Moro, and Chulupi (Matson et al., '68a). Other than this increased frequency of R" (CDE), Rh-results are not inconsistent with the data reported in other studies of South American Indians, which have been cited.

It should be noted that because of the

possibility of misclassification, pointed out by Layrisse, Layrisse and Gershowitz ( '70) , chromosome frequencies among American Indians for R' (CDE) may be misleadingly depressed. This is dependent upon a tech-nical problem with anti-rh' (anti-C) anti-sera of certain types which leads to mis-classification of RhJ lh j as R h j t h , and Rh.rh as Rh2rh. The order of magnitude of this error, as reported by Chown does not exceed 0.071.

The Mennonites' Rh-Hr chromosome frequencies were somewhat different from Western European Caucasians' frequen-cies. Their 0.22 frequency of r (cde) was significantly lower than that frequency of "d", 0.388, given by Mourant ( '54) for the Berne Swiss, of r (cde) for the Dutch, and many other European groups. The Men-nonite frequency for R' (CDe) was some-what higher than the corresponding fre-quencies for both groups of Hutterites, in Steinberg et al.'s ( '67) study. The pheno-typic incidence of ' Rh-positivity ( 9 0 % ) was somewhat higher than that of Jackson et al. ( '68) for Indiana Amish (32% ). The phenotypic frequencies of the Mennonites were quite similar to McKusick's figures for the Pennsylvania Amish, both, for exam-ple, showing the absence of Rh2Rh2 with significant and high frequencies for RhiRhi, RhiRh2, and rh. The latter in-cidence was a good deal higher in the Pennsylvania group (0.275) than in any of the related Anabaptists, and was thus typical of the deme only.

Kelt group. The absence of Kell-posi-tive individuals among the Indians tested is typical of American Indian populations. Although Pantin and Junqueira ( '52) found 23.3% of 73 Brazilian Indians to be Kell-positive, Chown and Lewis ( '57) have argued that the presence of the K antigen in North American Indians or Eskimos indicates Caucasian admixture. No puta-tively pure American Indian population ever tested for Kell and Cellano antigens has shown homozygous K, and very few have heterozygous Kk, and the Pantin and Junqueira work is a notable exception in that regard (Matson et al., '68a; Allen, '59; Salzano, '61). Caucasians' frequencies for k are also generally high, but rarely 1.0 (Mourant, '54).

Duffy group. Only two groups showed

GENETIC STUDIES IN PARAGUAY 3 3 9

homogeneity for the Duffy antigen; the Guayaki and the Tapiete were all positive for Fy\ Other estimated gene frequencies among the Indian groups were above 0.67. This is the expected range for Amerinds, and the frequency of 1.0 has not been re-ported often. The Mennonites' result of 0.46 for Fj/° is just above the 0.41 seen in Swiss and other western European Cauca-sians (Mourant, '54). McKusick's Amish had an Fya frequency of 0.56 in Lancaster County, Pa. (McKusick et al., '67). The frequency for the Indiana Amish was 0.546 (Jackson et al., '68).

Diego group. The interesting fact about our results for Di* is more its absence from five Indian groups than its presence in the rest. However, this phenomenon has occurred in other groups, as reported in two Brazilian tribes (Matson, Sutton, Pessoa, Swanson, and Robinson, '68b). It is a marker for persons of Mongoloid origin.

Serum globulins. The Gm and Inv fre-quencies are similar to those found in other Amerindian groups (Salzano and Stein-berg, '65; Salzano et al., 7 3 ) . The intra-Guarani relationship has already been discussed.

Inv'. Frequencies among South Ameri-can Indians range from 0.19 to 0.76 in data from previously published work, reviewed by Salzano et al. ( '73) . Our data in that regard are not unusual, except for some extremely low values for the Guayaki bands, a dramatic 0.12 for the Ache Kwera and for Cheroti (0 .09) . The Ache Kwera also have a very low frequency of Hp1. The Chulupi Inv1 value is also low, 0.20.

Serum haptoglobins. The range for the frequency of Hp 1 in South American Indians is 0.21 to 0.90, both Venezuelan figures (Arends and Gallango, '62, '64). The low incidence for the Yupa tribe is only very slightly lower than our computed frequency for the Guayaki. When consid-ering the Ache Kwera as a separate, endo-gamous band, which they were until only recently (Brown and Gajdusek, , ' 69b) , their Hp 1 gene frequency becomes the lowest reported for South American In-dians. Similar data have been reported by Matson et al. ( '69) , but without the band breakdown for the Guayaki.

Sources. Other than early works in

studying ABO distribution, the only studies of Paraguayan or Chaco Indians are the works of Matson et al. ( '68a, '69), and a study of H-Le" salivary secretion of Maca Indians by Salzano, Moreno, Palatnik and Gershowitz ( 7 0 ) . The earlier ABO sources are listed above.

ACKNOWLEDGMENTS

An expedition of this magnitude would be impossible without the help of many individuals.

For his inspiring ideas and important expertise, our thanks to the late Dr. Alfred Metraux; and to Dr. Jean Guiart, who arranged a meeting with Pierre and Helen Clastres and with Lucien Sebag, our expert guides and friends during the visit to the Guayaki Indians and to the Chaco. To Don Manuel Pereira for his aid on our mission to the Guayakis.

To officials of the government of Para-guay, who allowed this mission to be con-cluded successfully,-particularly Sr. Juan Alfonso Borgognon, Director of the De-partamiento de Asuntos Indigenos; and to Dra. Branka J. Susnik of the Museo Etno-grafico del Paraguay, for her helpful infor-mation concerning indigenous groups.

To the many officials of the U. S. govern-ment and U. S. Embassy in Asuncion for helping with technical and legal arrange-ments, travel, meetings and logistics, especially Dr. Eugene McCarthy, Director of the Servicio Cooperativo Interamericano de Salud Publica; Dr. Clifford A. Pease, and Mr. William P. Snow.

To the officials of the French Embassy in Asuncion, in helping us to contact those French anthropologists named above.

To Mr. Roy Wiggins, Catholic Relief Or-ganization in Asuncion, who helped ar-range our trip to the Moro (Ayore) Indians; and to Father Bruno Stella at the Catholic Mission Camp Maria Ausiliodora, where we met the Moro. To Father Bulber and the other priests and seminarians of the Oblates of Mary Immaculate, under whose auspices the Marescal Estigarribia mission to the Chulupi, Guarayu, and Guasurangue band of Tapiete Indians is run.

To the many persons associated with the Mennonite Colonies in the Chaco, who helped with our visit to Loma Plata and Chaco tribes; in particular, our thanks to

3 4 0 S. BROWN, GAJDUSEK, LEYSHON, STEINBERG, K. BROWN AND CURTAIN

Miss Ann Klassen and Dr. Labun, Dr. Willms, Dr. Wilhelm Kaethler and Mr. Frank Wiens; and to Mr. David Hein, head of the Yalve Sanga Mission of the Mennon-ites to the Lenguas, Tobas, and Chulupis.

To the South America Indian Mission and the New Tribes Mission for providing us with abundant background information on the Ayores.

We also thank Dr. Nicholas Escobar, Director of the Texaco Company in Colom-bia, and his staff at Orito for helicopter transport and other logistics support which he made available to use in the Putumayo region. We are grateful to Sr. Ugo Cuevas of Cali, Colombia for sea transport from Buenaventura to Belem on the Rio Docampado, and the return voyage in one of his shrimping vessels.

To all those associated with our labora-tories who have helped to prepare and edit data and the final manuscript, and in par-ticular, Marion Poms, Peggy Dawson and Dulcie Brown for their patient and fre-quent assistance.

LITERATURE CITED Alfred, B. M., T. D. Stout, M. Lee, J. Birkbeck

and N. L. Petrakis 1970 Blood groups, phos-phoglucomutase and cerumen types of the Anaham (Chilcotin) Indians. Am. J. Phys. Anthrop., 32: 329-338.

Allen, F. H., Jr. 1959 Summary of blood group phenotypes in some aboriginal Americans. Am. J. Phys. Anthrop., 17: 86.

Alvarez, A. G. 1939 Comprobaciones biologicas en aborigenes argentinos — consideraciones sobre los grupos sanguineos de los Matacos. Comision Honoraria de Reducciones de Indios, Buenos Aires, 6: 25-34 .

Arends, T., and M. L. Gallango 1960 Hapto-globin and transferrin groups in Venezuela. Eighth Congress of the International Society of Blood Transfusion, Tokyo. Proceedings. (S. Karger, Basel, 1962) .

1962 Frequencies of haptoglobin types in various South American populations, Mexico. Proceedings. (S. Karger, Basel, 1964)

Beals, C. 1961 Gos: God of the Chaco. In: Nomads and Empire Builders, pp. 124-143. Chilton Company, Philadelphia and New York.

Belaieff, J. 1946 The present day Indians of the Gran Chaco. In: Bureau of American Ethnology, Bull. No. 143. Handbook of South American Indians. Julian H. Steward, ed. Smithsonian Institution, Washington, D.C. (Re-published, Cooper Square, New York, 1963) , 1: 371-380.

1958 The Indian as I met him in the Chaco. Miscellanea Paul Rivet Octogenario Dictata (2 vols.) . (Proceedings of the 8th In-

ternational Congress of Americanists) , 2: 43-66 , 1958. (Mexico)

Bernstein, F. 1930 Fortgesetzte Untersuchungen aus der Theorie der Blutgruppen. Ztschr. fur induktiv. Abstammungs und Vererbungslehre, 56: 233-273.

Bertoni, M. S. 1941 Los Guayakies. Revista de la Sociedad Cientifica del Paraguay, 2: 1-62.

Brinton, D. G. 1898 The linguistic cartography of the Chaco region. Proc. American Philoso-phical Society, 37: 178-205.

Brown, K. S., and R. S. Johnson 1970 Popu-lation studies of Southwestern Indian tribes. III. Serum protein variations of Zuni and Papago Indians. Human Heredity, 20: 281-286.

Brown, S. M., and D. C. Gajdusek 1969a In-fanticide, intentional abortion, and live burial in a murderous band of honey and salt gather-ing nomadic Chaco Indians: Human ecology of the Ai'yore (Moro) Indians. Program and Abstracts, 79th Annual Meeting of the Ameri-can Pediatric Society, Atlantic City, April 3 0 -May 3, p. 27.

1969b Population control by child sacrifice and the teaching of cannibalism by children in a nomadic, polyandrous, hunting-gathering group; Guayaki Indians of Paraguay. Program and Abstracts, 79th Annual Meeting of the American Pediatric -Society, Atlantic City, April 30-May 3, p. 28.

Cadogan, L., and M. de Colleville 1963 Les Indiens Guayaki de L'Ynaro (Paraguay) (Premiere Partie). Bulletin de la Faculte des Lettres de Strasbourg, 41 (Tilas III): 439-458.

1964 Les Indiens Guayaki de L'Ynaro (Paraguay) (Seconde Partie). Bulletin de la Faculte des Lettres de Strasbourg, 41 (Tilas IVy. 499-532.

Campana, D. D. 1903 Contributo all'etnografia , dei Toba. Archivio per l'Antropologia e la Etnologia, Florence, 33: 287-322.

Chown, B„ and M. Lewis 1957 The Kell anti-gen in American Indians. Am. J. Phys. An-throp., 15: 149-156.

Chown, B„ M. Lewis and H. Kaita 1965 The Duffy blood group system in Caucasians: evi-dence for a new allele. Am. J. Hum. Genet., 17: 384-389.

Clastres, P. 1964 Compte rendu de mission chez les indiens Guayaki (Paraguay). L'Homme, Revue Fran?aise de Anthropologic publiee par l'Ecole Pratique des Hautes Etudes, Sorbonne, 4: 122-125.

Clastres, P., and L. Sebag 1963 Cannibalisme et mort chez les Guayakis. Revista de Museu Paulista, Sao Paulo, n.s. 14: 174-181.

Colleville, M. de 1972 Personal communica-tion, based upon his conversation with Dra. Branka Susnik of the Museo Etnografico de Historia Natural, Asuncion.

Coryn, A. 1922 Los indios Lenguas, sus cos-tumbres y su idioma, con compendio de gramatica y vocabulario. Anales de la Sociedad Cientifica Argentina, Buenos Aires, 93: 2 2 1 -282.

Curtain, C. C. 1962 Separation of haemo-globins Lepore and H from A and A2 in starch

GENETIC STUDIES IN PARAGUAY 3 4 1

or acrylamide gels buffered with tris-E.D.T.A. J. Clin. Path., IS: 288-289.

Curtain, C. C., C. Kidson, D. C. Gajdusek and J. G. Gorman 1962 Distribution pattern, population genetics and anthropological sig-nificance of thalassemia and abnormal hemo-globins in Melanesia. Am. J. Phys. Anthrop., 20: 475—483.

D'Orbigny, A. D. 1839 L'homme americain de l'Amerique meridionale. Pitois-Levrault, Paris.

Fretz, J. W. 1953 Pilgrims in Paraguay. Herald Press, Scottsdale, Pa.

Gajdusek, D. C. 1963 Paraguayan Indian Ex-peditions to the Guayaki and Chaco Indians, August 25 to September 28, 1963. National In-stitute of Neurological Diseases and Stroke, National Institutes of Health, Bethesda.

Grubb, W. B. 1914 An unknown people in an unknown land: an account of the l i fe and customs of the Lengua Indians of the Para-guayan Chaco, with adventures and experi-ences during twenty years' pioneering and ex-ploration amongst them. 4th edition. Seeley, Service and Co., Ltd., London.

Gurevitch, J., and E. Margolis 1963 Blood groups among Kurdistani Jews. Ann. Eugenics, London, 18: 94-95.

Hawtrey, S. H. C. 1901 The Lengua Indians of the Paraguayan Chaco. J. Roy. Anthrop. Inst., London, 31: 280-299.

Jackson, C. E., W. E. Symon, E. L. Pruden, I. M. Kaehr and J. D. Mann 1968 Consanguinity and blood group distribution in an Amish iso-late. Am. J. Hum. Genet., 20: 522-527.

Janzen, A. E. 1944 Glimpses of South America. Mennonite Brethren Publishing House, Hills-boro, Kansas, 130 pp.

— — 1962 The Moro's spear. Board of Mis-sions of the Conference of the Mennonite Brethren Church, Hillsboro.

Karsten, R. 1923 The Toba Indians of the Bolivian Gran Chaco. Acta Acad. Aboensis (Abo, Finland) , 4: 1-126.

Kelm, H. 1960 Zur Frage der ethnographischen Einordnung der Ayore, Moro und Yanaigua im Ostbolivianischen Tiefland. Baessler-Archiv, n.s. 8: 335-361.

1963 Die Zamuco (Ostbolivien). Z. Ethnol. (Berl in) , 88: 66-85.

Kersten, L. 1905 Die Indianerstamme des Gran Chaco bis zum Ausgange des 18 Jahrhunderts. Ein Beitrag zur historischen Ethnographie Siidamerikas. Inter. Arch. Ethnographie, Leiden, 17: 1-75.

Key, H., and M. Key 1961 Bolivian Indian tribes: Classification, bibliography, and map of present language distribution. Summer Institute of Linguistics, Publications in Linguistics and Related Fields, No. 15, Norman, Oklahoma.

Koch-Griinberg, T. 1902 Die Maskoi-Gruppe im Gran Chaco. Mitteilungen der Anthropologi-schen Gesselschaft, Vienna, 32: 130-148.

1903 Die Guaikuru-Gruppe, Mitteilungen der Anthropologischen Gesellschaft, Vienna, 33: 1 - 1 2 8 .

k ' n r n V i f r r r A o r » H R T H n r w l f o r i n t ; - n 1 , .

zymology, S. P. Colowick and N. O. Kaplan, eds. Academic Press, New York, 1: 323-327.

Kurczynski, T. W., and A. G. Steinberg 1967 A general program for max imum likelihood estimation of gene frequencies. Am. J. Hum. Genet., 19: 178-179.

La Hitte, C. de, and H. F. C. ten Kate 1897 Notes ethnographiques sur les Indiens Guaya-quis et description de leure caxacteres physi-ques. Anales del Museo de La Plata, Vol. 2.

Layrisse, Z., M. Layrisse and H. Gershowitz 1970 Blood typing studies in American Indians: mis-classification of R, phenotypes. Am. J. Phys. Anthrop., 32: 465-470.

Lehmann-Nitsche, R. 1908 Estudios antropolo-gicos sobre los Chiriguanos, Chorotes, Matacos, y Tobas (Chaco occidental). Anales del Museo de la Plata, n.s. 1: 53-149.

1923 Mitologia sudamericana VI. La astronomia de los Tobas. Revista del Museo de La Plata, 27: 267-285.

1936 Die sprachliche Stellung der Choropi (Gran Chaco). Z. Ethnol. (Berl in) , 68: 118-124, 303-304.

Loukotka, C. 1931 Vocabularios ineditos o pocos conocidos de los idiomas Camakoko, Sanapana, Angaita, y Sapuki. Revista del Instituto de Etnologia de la Universidad Nacional de Tucu-man, 1: 557-592.

Lozano, P. 1873-74 Historia de la conquista del Paraguay, Rio de la Plata y Tucuman, Andres Lamas, ed. Biblioteca del Rio La Plata, Buenos Aires.

Matson, G. A., and H. F. Schrader 1933 Blood grouping among the "Blackfeet" and "Blood" tribes of American Indians. J. Immun., 25. 155-163.

Matson, G. A., J. Swanson and A. Robinson 1966b Distribution of hereditary blood groups among Indians in South America. III. In Bolivia. Am. J. Phys. Anthrop., 25: 13-33.

Matson, G. A., H. E. Sutton, J. Swanson and A. Robinson 1966a Distribution of heredi-tary blood groups among Indians in South America. II. In Peru. Am. J. Phys. Anthrop., 24: 325-349.

Matson, G. A. 1968a Distribution of hereditary blood groups among Indians in South America. VI. In Paraguay. Am. J. Phys. Anthrop., 29. 81-98.

1969 Distribution of hereditary blood groups among Indians in South America. VII. In Argentina. Am. J. Phys. Anthrop., 30: 61-84.

Matson, G. A., H. E. Sutton, E. M. Pessoa, J. Swan-son and A. Robinson 1968b Distribution of hereditary blood groups among Indians in South America. V. In Northern Brazil. Am. J. Phys. Anthrop., 28: 303-330.

Mayntzhusen, F. C. 1925 Guayaki-Forschungen. Z. Ethnol. (Berl in) , 57: 315-318.

Mazza, S., and I. Franke 1927a Grupos san-guineos de indios y de auctoctonos del norte argentino (nota preliminar). La Prensa Med. Argentina, 14: 408.

1927b Groupos sanguineos de indios y

3 4 2 S. BROWN, GAJDUSEK, LEYSHON, STEINBERG, K. BROWN AND CURTAIN

McKusick, V. A., J. A. Hostetler and J. A. Egeland 1964a Genetic studies of the Amish. Back-ground and potentialities. Bull. Johns Hopkins Hosp., 115: 203-222.

McKusick, V. A., J. A. Hostetler, J. A. Egeland and R. Eldridge 1964b The distribution of certain genes in the old order Amish. Cold Spring Harbor Sympos. Quantit. Biol., 29: 9 9 -114.

McKusick, V. A., W. B. Bias, R. A. Norum and H. E. Cross 1967 Blood groups in two Amish demes. Humangenetik, 5: 36-41 .

Metraux, A. 1929 Un ancien document peu connu sur les Guarayu de la Bolivie orientale. Anthropos, 24: 913-941.

1937 Etudes d'ethnographie Toba-Pilaga (Gran Chaco). Anthropos, 32: 171-194, 3 7 8 -401.

1946 Ethnography of the Chaco. In: Bureau of American Ethnology, Bull. No. 143, Handbook of South American Indians. Julian H. Steward, ed. Smithsonian Institution, Wash-ington, D.C. (Republished, Cooper Square, New York, 1963), 1: 197-370.

1948 Tribes of eastern Bolivia and the Madeira headwaters, the Guarayu and Pau-serna. In: Bureau of American~Ethnology, Bull. No. 143, Handbook of South American Indians. Julian H. Steward, ed. Smithsonian Institution, Washington, D. C. (Republished Cooper Square, New York, 1963) , 3: 430-438.

1949 Warfare, cannibalism, and human trophies. In: Bureau of American Ethnology, Bull. No. 143, Handbook of South American Indians. Julian H. Steward, ed. Smithsonian In-stitution, Washington, D.C., 5: 383-409.

Mourant, A. E. 1954 The distribution of the human blood groups. Charles C Thomas, Spring-field, Illinois.

Nordenskiold, E. 1913 Indianer leben, El Gran Chaco (Sudamerika). G. Nerseburger, Leipzig.

1917 The Guarani invasion of the Inca empire in the sixteenth century: an historical Indian migration. Geogr. Rev., 4: 103-121.

—> 1919 An ethno-geographical analysis of the material culture of two Indian tribes in the

•Gran Chaco. Comparative Ethnographical Studies, I: 1-293. -

Norwood, E. B. 1962 The fullness of time for the Guiday Gosode. Good Neyvs Broadcaste (Back to the Bible Broadcast. Monthly Supple-mentary Publicatiori, Lincoln, Nebraska), 20: 12-15. •' . _

O'Leary, T. J. 1963 Ethnographic bibliography of South America. Human Relations Area File, New Haven, Connecticut.

Palavecino, E. 1939 Relevamiento antropome-trico de un Indio Ashlushlai. Notas del Museo de La Plata, Universidad Nacional, 4: 413-414.

Pantin, A. M., and P. C. Junqueira 1952 Blood groups of Brazilian Indians. Am. J. Phys. Anthrop., 10: 395-405.

Pencille, W. 1960 Operation — Guiday Gosode. Amazon Valley Indian. (South America Indian Mission, West Palm Beach, Florida), 55 (10 ) : 4-5 .

1961a Uejai, saved three times. Ama-

zon Valley Indian. (South America Indian Mis-sion, West Palm Beach, Florida), 5 6 ( 2 ) : 1 -3

1961b Bolivia — Guiday Gosode. Ama-zon Valley Indian. (South America Indian Mis-sion, West Palm Beach, Florida), 5 6 ( 8 ) : 1-3.

1961c Bolivia — Guiday Gosode. Ama-zon Valley Indian. (South America Indian Mis-sion, West Palm Beach, Florida), 5 6 ( 9 ) : 3-4 .

1967 Bridging the gap from stone age to space age. Evangelical Missions Quarterly. (Evangelical Missions Information Service, Springfield, Pennsylvania) , 4 ( 1 ) : 15-26.

Pencille, W., and H. Pencille 1960 Paganism in the raw. Amazon Valley Indian. (South America Indian Mission, West Palm Beach, Florida), 5 5 ( 1 2 ) : 1 -3 .

1963 Ayoreo culture. Unpublished man- • uscript, San Jose de Chiquito, Bolivia, and South America Indian Mission, Lake Worth, Florida, 104 pp.

* Pierini, F. 1910 Mitologii de los Guaiayos de Bolivia. Anthropos, 5: 703-710.

Pollitzer, W. S„ R. C. Hartmann, H. Moore, R. E. Rosenfield, H. Smith, S. Hakim, P. J. Schmidt and W. C. Leyshon 1962 Blood types of the Cherokee Indians. Am. J. Phys. Anthrop., 20: 33—43.

Pollitzer, W. S., D. S. Phelps, R. E. Waggoner and W. C. Leyshon 1967 Catawba Indians: Mor-phology, genetics and history. Am. J. Phys. Anthrop., 26: 5-14 .

Ribeiro, L. 1934 Les grpupes sanguins chez les Indiens Guaranis du Bresil (The blood groups of the Guarani Indians of Brazil). C. R. Soc. Biol. (Paris) , 117: 777-778.

Ribeiro, L„ W. Berardinelli and M. Roiter 1935 Grupo sanguineo dos indios guaranys (Blood groups of the Guarani Indians). Arquivos de medicina legal e identificacao (Rio de Janeiro), 5 ( 1 1 ) : 59-65 .

Rosen, E. C. G. B. von 1906 The Chorotes Indians in the Bolivian Chaco: A preliminary report dedicated to the ,14th International Congress of Americanists at Stuttgart, 1904. (Internationaler amerikanisten-kongress: vier-zehnten tagung, Stuttgart*. 1904). W. Kohl-hammer, Stuttgart, 2: 649-658.

Salzano, F. M. 1957 The blood grotips -of South American Indians. Am. j. Phys. Anthrop., 15: 555-579. .. "*'. > / f ' s S '

- — r — 1961 Studies ori the Caingang lndiaris. III. Blood groups. Am. ..J,: Phys. Anthrop., 19: 391-404. . . ".

Salzano, F. M., and A. G. Steinberg. 1965 The Gm and Inv groups of Indians from Santa Catarina, Brazil. Am. J. Hum. Genet., 17: 273-279.

Salzano, F. M., R. Moreno, M. Palatnik and H. Gershowitz 1970 Demography and H-Le* salivary secretion of the Maca Indians of Para-guay. Am. J. Phys. Anthrop., 33: 383-388.

Salzano, F. M., A. G. Steinberg and M. A. Tepfen-hart 1973 Gm and Inv allotypes of Brazilian Cayapo Indians. Am. J. Hum. Genet., 25: 167-177.

Schmidt, M. 1936 Los Guarayu. Revista de la Sociedad Cientifica del Paraguay, 3. 158-194.

GENETIC STUDIES IN PARAGUAY 3 4 3

Shreffler, D. C., and A. G. Steinberg 1967 Fur-ther studies on the Xavante Indians. IV. Serum protein groups and the SCi trait of saliva in the Simoes Lopes and Sao Marcos Xavantes. Am. J. Hum. Genet., 19: 514-523.

Smith, W. 1950 Paraguayan Interlude. Herald Press, Scottsdale, Pa.

-y: Snethlage, E. H. 1935 Nachrichten iiber die Pauserna-Guarayu, die Siriono am Rio Baures und die S7 Simonianes in der Nahe der Serra S. Simon. Z. Ethnol., 67: 278-293.

Sorenson, E. R., and D. C. Gajdusek 1966 The study of child behavior and development in primitive cultures: A research archive for ethnopediatric film investigations of styles in the patterning of the nervous system. Pediatrics, 37(Part II) (Suppl . ) : 149-243.

South America Indian Mission, Inc. n.d. The story of Zapoco. South America Indian Mission, West Palm Beach, Florida.

Steinberg, A. G. 1962 Progress in the study of genetically determined human gamma globulin types ( the Gm and Inv groups). Progr. Med. Genet., 2: 1 -33 .

Steinberg, A. G„ H. K. Bleibtreu, T. W. Kurczyn-ski, A. O. Martin and E. M. Kurczynski 1967 Genetic studies on an inbred human isolate. In: Third International Congress of Human Ge-netics, Chicago, 1966. Proceedings. J. F. Crow, and J. V. Neel, eds. Johns Hopkins Press, Balti-more, pp. 267-289.

Steward, J. H„ ed. 1946-59 Handbook of South American Indians. In: Bureau of Amer-ican Ethnology, Bull. No. 143. Smithsonian In-stitution, Washington, D.C.

Stewart, T. D. 1952 A bibliography of physical anthropology in Latin America: 1937-1948. Wenner Gren Foundation for Anthropological Research, New York.

Susnik, B. J. 1961 Estudios Guayaki. Boletin de la Sociedad Cientifica del Paraguay y del Museo Dr. Andres Barbero Etnografico de Historia Natural, 5: 1-217.

1962 Vocabulario ace-Guayaki (con-tinuacion). Boletin de la Sociedad Cientifica del Paraguay y del Museo Dr. Andres Barbero Etnografico de Historia Natural, 6: 105-220.

1963 La lengua de los Ayoweos-Moros. Boletin de la Sociedad Cientifica del Paraguay y del Museo Etnografico, 8: 1-148.

Urizar, R. 1942 Grupos sanguineos de aucto-tonos del Chaco Paraguayo. America Indigena (Mexico City), 2: 49-50 .

Vellard, J. 1934 Quelques recherches sur les groupes sanguins chez des Indiens du Paraguay. C. R. Soc. Biol. (Paris ) , 116: 4 7 1 ^ 7 2 .

W. H. O. Scientific Group on Genes, Genotypes, and Allotypes of Immunoglobulins 1965 Notation for genetic factors of human im-munoglobulins. Bull. WHO, 33: 721-724.