Paraaitology (1974), 68, 117-121:; 1717 With 4 plates

Further parasites of the family Garniidae (Coccidiida: Haemosporidiidea) in Brazilian lizards. Fallisia effusa gen.nov., sp.nov. and Fallisia modesta gen.nov., sp.nov. R. LAINSON,* IRENE LANDAUt and J. J. SHA W*

(Receivecl10 August 1973)

SUMMARY FaUisia effusa gen.nov., sp.nov. (Haemosporidüdea: Garniidae) is de- scribed iI;Lthe teiid lizard, N eusticurus bicarinatus, from Pará State, north Brazil. Schizogony and gametogony both take place within thrombocytes and white cells of the peripheral blood. There is no development in the red blood cells, by which character the organism is difIerentiated from Garnia, the other known genus within the family. Another species,FaUisiamodesta gen.nov., sp.nov., is described in the iguanid lizard, Tropidurus torquatus hispidus, algo from Pará, Brazil. It is distinguished from F. effusa by its development principally in the lymphocytes, as opposed to the thrombo- . cytes. There are other, striking morphological differences between the two species. It is recommended that great care is needed in the interpretation of so-called 'exo-erythrocytic stages' of saurian plasmodia, which may well really belong to members of the Garniidae, in mixed infections.

INTRODUCTION Recently (Lainson, Landau & Shaw, 1971) we erected a new sporozoan fami1y, Garniidae (Coccidiida: Haemosporidiidea), for the inclusion of some newly dis- covered, non-pigmented haemosporidia of South American lizards. These parasites undergo schizogony and dimorphic gametogony in the cells of the peripheral blood of the vertebrate host but, when development is in the redblood cells, there is no production of malarial pigmento By these characters the parasites are differ- entiated from those of the existing three families of the suborder Haemospori- diidea: from the Plasmodüdae and the Haemoproteidae by the absence of pigment, and from the Leucocytozoidae and the Haemoproteidae by the asexual cycle in the circulating blood. The type genus Garnia is characterized by its erythrocytic schizogony and gametogony within immature and mature red blood cells of the peripheral blood, with no production of malarial pigmento In some species there is developed a variable number of azurophilic granules in the cytoplasm, but these are negative to polarized light. Exoerythrocytic schizogony occurs in lymphocytes, monocytes and throm bocytes. Known speciesinclude Garnia gonatodi (Telford, 197O) Lainson, * The WellcomeParasitology Unit, Caixa Postal 3, Belém Pará, Brazil. t MuseumNational d'Histoire Naturelle, Laboratoire de Zoologie(Vers), 57 Rue Cuvier, Paris Ve, France. 8-2 118 R. LAINSON, IRENE LANDAU AND J. J.. SHAW

Landau & Shaw, 1971; G. morula (Telford, 1970) Lainson, Landau & Shaw, 1971; G. telfordi Lainson, Landau & Shaw, 1971; and G. utingensis Lainson, Landau & Shaw, 1971. When these parasites were first examined by the electron microscope, minute cytoplasmic inclusions were thought to be sub-light-microscopic grains of pigmento Subsequent opinion, however, is that this was not so, and that we can confidently reiterate our previous definition that the parasites of this family do not produce malarial pigment when developing in the red blood cells. The bulk of the present paper deals with the discovery of an extraordinary new haemosporidian of the teiid lizard Neusticurus bicarinatus (Linnaeus, 1758) Duméril & Bibron, 1839, from Ananindeua and Capanema, Pará State, north Brazil. Although the parasite is clearly related to Garnia, it undergoes both schizo- gony and gametogony in the thrombocytes and, more rarely, the lymphocytes. There is no development in the red blood cells, and we reeI that this peculiarity alone justifies separating the parasite from the genus Garnia within the family Garniidae. The name Fallisia gen.nov. is proposed, in honour of Professor Murray Fallis who has contributed much to our knowledge of other pigmentless haemosporidia, the Leucocytozoidae. A further species of this parasite is also described in the iguanid lizard Tropidurus torquatus hispidus (Spix, 1825) Burt & Burt, 1930, from Belém, Pará, Brazil.

MATERIAL AND METHODS N. bicarinatus is a small, semi-aquaticlizard of the family Teiidae. It lives in humid forested aroos, particularly along the banks of the rivers and streamsin which it rapidly seeks protection against would-be predators. The lizards were kept in glass aguaria, with the option of wet or dry conditions, and a number of loose stones under which these animaIs like to hide. T. torquatushispidus, on the other hand, is a sun-Iovingreptile. It prefers open country, waste land, roadsidesor the larger forest clearings.This lizard keepswell in dry aguaria, and water is neededonly for drinking purposes. The lizards were alI bled from the orbital sinus with a glass pipette drawn to a very fine point. Blood illms were dried quickly, fixed at once in absolute methyl alcohol, and stained for 1-2 h in Giemsa stain (30 drops to 15 ml distilled water buffered to pH 7.4).

DESORIPTION OF THE PARASITES Fallisia effusa gen.nov.,sp.nov. (PI. 1, figs. 1-15; PI. 2, figs. 16-19; PI. 3, figs. 29-40; PI. 4, figs. 41-46) In spite of a long study of blood films from many infected lizards and the examination of day-to-day developmentin individual animaIs, it has remained difficult to be sure of the complete evolution of the parasite in the peripheral blood. Thus, while it is simple to state that infection starts with a period of schizogonyand endswith an infection consistingalmost entirely of mature gameto- Haemosporidia01 lizards

cytes, it is not so easy to interpret the abundance of bizarre forros of the organism in between these two stages. Luckily the infection is extremely common in N. bicarinatus, enabling a large amount of material to be examined. Twenty-six out of thirty-two lizards examined were positive. The youngest parasites seen within the thrombocytes (76 %) or lymphocytes (14 %) measured only ,...,2.0 x 1.0 ",m (PI. 1, figo 3) and are clearly derived from mature, rupturing schizonts (PI. 1, figo 8; PI. 3, figo 34). Some of these merozoites give rise to further schizonts and others apparently develop directly into male and female gametocytes.

Schizogony The length of the initial schizogony cycle remains undetermined as, in the absence of experimental transmission, we have been unable to follow the infection from the very beginning. The young trophozoite had an irregular shape, often stellate, with tiny pseudo- podia. Nuclear division (PI. 1, figs. 4-8; PI. 3, figs. 29-34) gives rise to separate chromatin masses (PI. 3, figo 30) or, frequently, t? a network ofnuclear material studded with more intensely stained granules (PI. 1, figs. 5,6). With increasing nuclear division the parasite assumes a smoother outline, the final product being round or oval (PI. 1, figo 7): mature schizonts measure '" 10.0-14.0 p;m in diameter and produce from 50-150 merozoites. These tiny bodies are mostly tear-shaped to triangular (PI. 1, figo 8; PI. 3, figo 34) and measure '" 2.0 x 1.0 p;m: they enter new thrombocytes and lymphocytes to form further schizonts, or gametocytes.

Gametogony Young gametocytesbegin to appear early in the infection, when schizogonyis still at a relatively low leveI and when the infection is light. Someof the invading merozoitesdo not undergo nuclear division, but enlarge: whilst still very small they are readily recognizable as young male and female gametocytes (PI. 1, figs. 9, 10). When there is only one parasitein the cell, these early stagesare oval to bean-shaped,but may be distorted by pressure from adjacent parasites in multiple infections. Some already show a variable number of tiny azurophilic granules, such as seenin speciesof Garnia. The microgametocytehas the usual diffusely scattered nuclear material, often with a distinct nucleolus, and has an overall pink hue which contrasts strongly with the vivid blue cytoplasm of the macrogametocyte.The nucleus of the female parasite forros a well-defined area, often band-form: again, there is usually a conspicuousnucleolus. The normal shape of the mature gametocytesappears to be an elongateoval (PI. 1, figs. 11, 12)but they are frequently deformedby pressure againstthe host-cellnucleus, or againstone anotherin cells with multiple infections (PI. 1, figo 14; PI. 3, figo 40). The mature female measures'" 10.0x 6.0 p,m: the males are smaller, '" 8.0 x 5,0 p,m. Both sexesmay have 1-3 prominent vacuoles (PI. 3, figs. 39, 40); more usually there is a single vacuole, measuring up to 1.0 p,m. Counts of gametocytes,in three different lizards which showed only mature

119 R. LAINSON, IRENE LANDAU AND J. J. SHAW forms, gave the proportions ofmales to females as 107: 136,119: 149 and 121: 181. A iourth lizard, however, initially showed only 2: 98; and 1 week later it appeared to have only female parasites in the peripheral blood (out of 230 gametocytes counted). We are at a 10ssto explain this strange disappearance of the micro- gametocytes. As far as we could determine, the development time of the gametocytes is 7 to 10 days. M ultiple infection of tM host cells We have previousIy stressed that muItipIe invasion of the ce11sis a prominent feature ofthe genus Garnia. It is even more pronounced in Fallisia effusa. Thus, a study of lizards with light and presumabIy recent infections showed non-infected and infected thrombocytes in the proportion of 25:29. DoubIe infections were very rare and the parasites were aImost aIways trophozoites and young schizonts, derived from a sma11number of mature schizonts of the tyPe described above. Two days Iater the number of mature schizonts had increased and the proportion of non-infectedjinfected ce11swas now 13: 41. Numerous ce11salready contained two or three parasites and, on subsequent days, the number of infected ce11ssteadiIy increased until virtua.lIy 100% ofthe thrombocytes were parasitized with variabIe mixtures oftrophozoites, schizonts and deveIoping gametocytes (PI. 1, figo 15; PI. 2, 16-19; PI. 4, figs. 41-46). At this stage the picture becomes even more compIicated by what appears to be a second forro of schizont producing a very small number of merozoites. These seem to produce a mixture of further schizonts or gametocytes, a11 within the same unruptured host ce11. Other strange, tenuous forms are produced which apparentIy break up into separate masses containing variabIe numbers of nucIei (PI. 1, figo 15; PI. 2, figo 18). The presence, already, of merozoites, young schizonts and growing gametocytes from the preceding schizogony makes it difficuIt to decide on the fate of many of the parasites resuIting from this second form of division. It may be that many of them deveIop no further: indeed, when they occupy a ce11containing a mature schizont it seems most unlikeIy that they can reach maturity before the ce11 ruptures (PI. 4, figo 43). Whatever the case, the schizogony cycIe sIowIy abates, Ieaving thrombocytes and Iymphocytes containing only mature gametocytes. The resuIts of the extra- ordinary muItipIe invasion of the ce11sis sti11 we11reflected in the number of gametocytes per 0011,which varies from 1 to 5 (PI. 1, figs. 12-14; PI. 3, figs. 36-40).

E.ffects on the Mst cells Even when they are quite small the trophozoites, schizonts and gameto~ytes alI have a profound effect on the host cell, in particular the thrombocyte. The thrombocytes of N.. bicarinatus are oval to cylindrical bodies, averaging 13..0x. 5,,0p,m, and possessing a densely staining nucleus of '" 9,,0x 5.0 p,m. Th~ cytoplasm is highly vacuolated and almost colourless, even after lengthy staining. It isbounded, however, by a delicate membrane which stains bright pink (PI. 1, 120 Haemosporidiao/ lizards 121 figo 1). When the cells are unduly traumatized, as in thin blood films; the cytoplasm may be extruded and appears as a thin, frothy layer adhering to the outside ofthe cell wall (PI. 1, figo 2). This gives the false impression of a delicate membranous structure lying within the cytoplasm of the thrombocyte: this feature is stressed, here, becauseuntil we examined normal thrombocytes we were under the mistaken impression that the membrane was produced by the parasite. With entry and growth of the parasite the limiting membrane of the thrombo- cyte thickens remarkably, often to such an extent that the infected cell takes on a cyst-like appearance: this is particularly so in cells containing mature gameto- cytes (PI. 1, figs. 11-14; PI. 3, figs. 37-40). Maximum alteration in the shape ofthe cells is seen in those containing mature schizonts or gametocytes, and in thoSe with multiple infections: it is mainly manifested by a shift from elongate to spherical, the infected thrombocytes sometimes reaching' '" 16.0 x 14.0 p,m (PI. 1, figs. 11-1.4; PI. 3, figs. 35-40). A great many parasitized thrombocytes undergo the partial extrusion of con- tents, discussed above, giving the erroneous impression that the parasites are actively emerging from the cells (PI. 1, figo 14; PI. 3, figs. 35,39). The host cell nucleus undergoes relativeIy little change, but there is some enlargement and distortion when it is squashed to one end or side of the thrombocyte. Infected lymphocytes were uncommon but readily recognizable by their lack of the pink, hypertrophied cell wall. Sporogony Attempts to induce exflagellation of the male gametocytesin slide/coversIip preparations have so far failed. Attempts are in progressto obtain sporogonic stagesin a variety of blood-suckinginsects.

Bost specificity We have not attempted to infect other speciesof lizards by blood inoculation, but have found no natlÚ'al infections of F. e.ffusain any other kind of lizard from the garoeareas of forest.

Fallisia modesta gen.nov.,sp.nov. (PI. 2, figs. 20-28; PI. 4, figs. 47-52). This is recorded in a singIe specimenof T. torquatushispidus captured on the roadsidein Utinga, near Belém. The infection was light but was foIlowed,in bIood films, for 15 days untiI the animal died.

Schizogony F. ?rWdestais a small, neat parasite developingprincipally in the lymphocytes, more rarely the monocytes. A solitary female gametocyte was once seen in a thrombocyte. The schizonts(PI. 2, figs. 20, 21; PI. 4, figs. 47, 48) are delicatelittle bodiesand remarkably consistentboth in dimensionsand in the number of merozoitesthey produce. They are round to oval, ranging from 5.0 x 5.0 to 9.0 x 7,5 p,m,average 5.5 x 5.0 p,m. The most commonnumber of merozoitesproduced is 16, this being 122 LAINSON, IRENE LANDAU AND J. J. SHAW recorded in 17 out of 28 mature forms examined: the range is from 1.2to 24. The merozoites are similar to those of F. e.ffusa,measuring about 2.0 x 1.0 p,m, and most often being tear-shaped (PI. 2, figo 22; PI. 4, figo 52).

Gametogony The young gametocytesare soondistinguishable as malesand femalesby their individual staining reaction (PI. 2, figa. 24, 27) and they quickly develop very conspicuousazurophilic granules. These are particularly noticeable in the pale- coloured male. The mature gametocytesare round to oval (PI. 2, figa. 25, 26; PI. 4, figa. 49, 50) and there is little difference in size for the two sexes.The male averages7.0 x 5.5 pm (range 8.5 x 5.5 to 6.0 x 6.0 pm) and the female 7.5 x 5.5 pm (range 8.5 x 5.5 to 7.0 x 6.0 pm). There is no conspicuouscytüplasmic vacuole, as is almost always seenin F. effusa,and the female nucleustends to be round or oval rather than bana-formoBoth males and femalesoften showa distinct nucleolus.

M ultiple infection of the host cells As in F. effusa, there is a strange period of multiple infection of the host celIs. This results either from repeated invasion of a given celI by merozoites of the primary schizonts, or from the secondary type of schizogony as described in F. effusa. This process is not nearly so pronounced as in that parasite, however, and is mostly seen in 'smear-cells' (PI. 2, figo 28; PI. 4, figo 51). We cannot be sure of the true nature of these host cells, but they are probably monocytes: they clearly are exceedingly fra~e, and alI appear to become ruptured in making the blood films. The nucleus becomes smeared out into a thin pink layer, with the liberated parasites remaining in a little group by the nuclear remnants. The rupturing , smear-celIs' may liberate single schizonts, morphologically similar to those in the lymphocytes, or a mixture of mature and maturing parasites, particularly gametocytes (PI. 2, figo 28; PI. 4, figs. 51, 52). Up to the death of the lizard these appeared slightly smalIer than the mature gametocytes in the lympho- cytes, but we reei that this is simply a question of immaturity.

Effects on lhe host cells The nucleus of the infected lymphocytes is strongly indented by the larger parasites, particularly the mature gametocytes, and the cytoplasm becomes virtually non-existent. We have already discussedthe infected 'smear-cells': the rupture of these cells cannot be regardedas purely due to the parasite, how- ever, as they are seenquite abundantly in noI:l-infectedTropidurus.

Sporogony:host specificity No information. DEFINITIONS 1. Fallisia gen.nov. With the general characters as previously given for the family Garniidae (Sporozoa:Coccidiida: Haemosporidiidea).Both schizogonyand gametogonyin

R. H aemosporidia of lizards 123 thrombocytes and white cenaof the circulating blood. No developmentin the red blood cena,by which characterthe parasiteis differentiated from Garnia,the other known genus of the family.

2. Fallisia effusa sp.nov. Principally parasitizingthe thrombocytes,more rarely the lymphocytes.Mature forms causing rounding and enlargementof the cell, displacementand flattening of the nucleus,and a peculiar hypertrophy of the thrombocyte waJIto give a cyst- 1ike appearance. Young trophozoites irregular in shape and moderately amoeboid. Schizonts becomingovoid or round towards maturity, whenthey are 10.0-14.0p,m in diameter and produce 50to 150tiny merozoites:merozoites tear-shaped and ,..,2.0 x 1.0p,m. Multiple invasion of the host celIsvery pronounced: secondary,smaller schizonts produce a mixture of asexual and sexual stages within the unruptured cell. Presence,algo, of many bizarre and possibly aberrant forms. Immature gametocytes appear early on in the infection and throughout the period of schizogony: they are quickly differentiated into microgametocytesand macrogametocytesby the overall pink colour of the male and the blue staining cytoplasm of the female. Mature gametocyteselongate-oval, averaging 10.0x 6.0 p,m (female)and 8.0 x 5.0 p,m (male). Both sexeswith one to three prominent, cytoplasmic vacuoles (usually one) and variable numbers of small azurophilicgranules. Older infections showing only gametocytesin circulating blood, often from one to five per cell, due to multiple infection of cells in the earlier stages of the infection. ROST.The lizard (Teüdae)Neusticurus bicarinatus (Linnaeus, 1758)Duméril & Bibron, 1839. LoCALITY. Riverine, tropical rain-forest: Ananindeua and Capanema,Pará State, north Brazil. TYPE MATERIAL.The Wellcome Parasitology Unit, The Instituto Evandro Chagas,Belém, Pará, Brazil, and the Museum of Natural History (Vers), Paris, France.3. Fallisia modesta sp.nov. Principally in the lymphocyteSof the circulating blood, very rarely the thrombo- cytes. Mature forms stongly indenting the host nucleus. Schizonts small and compact, round to oval, from 5.0 p,m to 9.0 x 7.5 p,m in diameter, average 5.5 x 5.0 p,m. Number of merozoites 12-24, most usually 16: tear-shapedand ,..,2.0 x 1.0 p,m. Multiple invasion of host cells, but nót so pronounced as in F. e.ffusa. Production of mixtures of schizonts and gametocytesin so-called 'smear-cells' which inevitably rupture in the preparation of blood films. Immature gametocytesappearing throughout period of schizogonyand are early on distinguishedas malesand femalesby their different stamingreaction. Mature gametocytes round or oval, averaging 7.5 x 5.5 p,m (female) and 7.0 x 5,5 p,m (male). No obvious cytoplasmic vacuole as in F. e.ffusa,but azurophilic granules ~ell developedand especiallyprominent in the male. 124 LAINSON, IRENE LANDAU AND J. J. SHAW

ROST.The lizard (Iguanidae) Tropidurus torquatus hispidus (Spix, 1825) Burt & Burt. 1930. LOCALITY. Roadside, near lake: Utinga, near Belém, Pará, north Brazil. TYPE MATERIAL. The Wellcome Parasitology Unit, the Instituto Evandro Chagas, Belém, Pará, Brazil, and the Museum <;>fNatural Ristory (Vers), Paris, France.

DISCUSSION Lizards ofthe New World are súrprisingly common1yinfected with pigmentless haemosporidia,and the infections are found in a wide range of different animaIs, in different habitats. There are now recordedone new genus,Saurocytozoon in the family Leucoc~ozoidae (Lainson & Shaw, 1969), four speciesof Garnia in the family Garniidaeand, now, two speciesof the newgenus Fallisia in the samefamily. Further speciesof the latter two parasitesare at presentbeing studied and wi11be describedin the near future. We mention this to stressthe remarkable fact that these interesting parasites should have remained undetected (or unrecognized)for so long, in spite of the active interest of numerous workers in saurian Plasnwdium species,throughout the New World, over a considerablenumber of years. It is early yet to theorize with any great confidenceon the relationship that the Garniidae may have with other, better known membersof the Haemosporidiidea, especiallyas we know so little about their sporogonyand vectors. At presentwe can but suggestthatthey form an evolutionary 1ink betweenthe Plasmodiidaeon the one hand (Garnia)and the Leucocytozoidaeon the other (Fallisia). The existenceof this group of parasitesraises some doubts as to previous inter- pretation of so-càlIed 'exo'"er~hroc~ic stages' of many of the known saurian Plasmodiumspecies. Unhappily it has generalIybeen assumed that any schizogonic process within white celIs or thrombocytes belonged to the Plasmodium being described.We have common1yfound lizards with mixed infections of Plasnwdium, Garnia and/or Fallisia, and clearly great caro must be taken before conclusions are drawn -preferably after repeated examination of blood films from selected animaIs over long periods. As with the Plasmodiidaeof lizards, we are very ignorant of the natural vectors of the Garniidae. It is remarkable, however, that infections are found in widely different speciesin very different ecologicalniches. This suggeststhat different vectors may be involved, although we do strongly suspectthe ubiquitous Culi- cOides,species of which seemto inhabit aJmostalI habitats: mosquito speciesalgo remain natural suspects,of course. Phlebotomid s~ndfliesare possiblevectors of some malarial parasites of lizards innorthAmerica (Ayala & Lee, 1970).In ourareaofBrazil, however, agreat many infected lizards are to be found where sandfliesdo not occur -in private gardens within the city of Belém, for example. In the meantime we can but hope that continuing attempts to produce sporo- gony in experimental insects may eventualIy bear fruit and lead to a better understanding of these organisms.

R. Parasitology, V 01. 68, Pare 2 Plate 1

1 Parasitology, VoZ. 68, Pare 2 Plate 2

LAINSON. J.RENE LANDAU ANDJ. J. SHAW

R. ParasitoZogy, VoZ. 68, Pari 2 Plate 3

R. LAINSON, IREKE LANDAU AND J. J. SHAW Parasitology, V 01. 68, Pare 2 Plate 4

R. LAINSON, IRENE LANDAU AND J. J..SHAW Haemasparidia aI lizards 125 The cQst ofthe colour plates was defrayed by the Wellcome Parasitology Unit,Instituto Evandro Chagas, Belém, Pará, Brazil.

We are most grateful to Dr William E. Duellman, Curator, Division of Her- petology, the University of Kansas, for the identification of the lizards.

REFERENCES AYALA, S. C. & LEE, D. (1970). Saurian malaria: development of sporozoites in two species of phlebotomine sandflies. Science 167, 891-2. LAINSON,R. & SHAW, J. J. (1969). A new haemosporidian of lizards, Saurocytozoontupinambi gen.nov., sp.nov., in Tupinumbus nigropunctatus (Teüdae). Parasitology 59, 159-62. LAINSON,R., LANDAU,I. & SHAW,J. J. (1971). On a new family ofnon-pigmented parasites in the blood ofreptiles: Garntidae faro. novo (Coccidiida: Haemosporidüdea). Some species of the new genus Garnia. lnternational Journal oi Parasitology 1, 241-50.

EXPLANATION OF PLATES PLATE 1 Figa. 1-15. FaUisia eff118agen.nov., sp.nov., from the lizard NeUBticurus bicarinatUB,as seen in the thrombocytes of Giemsa-stained blood films. 1, normal thrombocyte; 2, traumatized thrombocyte, showing partial extrusion of cytoplasm; 3, young trophozoite; 4-7, primary schizogony in early infection, note partia! extrusion of cell contenta in some instances; 8, ruptured, mature primary schizont; 9, 10, developing microgametocyte and macrogameto- cyte; 11-14, mature maIe and female gametocytes, showing hypertrophy of the host cell walI, multiple infections and partia! extrusion of the cell contents; 15, multiple infection with a. microgametocyte, young schizonts and a number of trophozoites.

PLATE 2 Figs. 16-19. Fallisia effusagen.nov., sp.nov., in the lizard Neusticurus bicarinatus. Multiple infection of thrombocytes with developing schizonts and gametocytes, and products of secondary schizogony. Figs. 20-28. Fallisia modestagen.nov., sp.nov., in the lizard Tropidurus torquatus hispidus. 20,21, primary schizogony in a lymphocyte and a monocyte; 22, mature, ruptured primary schizont; 23, 24, young female and male gametocytes in lymphocytes; 25, 26, mature female and male gametocytes; 27, multiple infection of a lymphocyte with trophozoites, young macrogametocyte and two mature male gametocytes; 28, 'smear-cell' with liberated schizont and two developing macrogametocytes. PLATE 3Figa. 29-40. Fallisia effUBagen.nov., sp.nov., in the lizard NeUBtwurUBbicarinatUB. 29-34,primary schizogony in the thrombocytes; 35, two young gametocytes, partial1y extruded from the thrombocyte; 36-40, developing and mature gametocytes, in variable numbers, inthrombocytes. Note hypertrophy of cell wall and prominent cytoplasmic vacuoles of thegametocytes.

PLATE 4 Figs. 41-46. Fallisia effusa gen.nov., sp.no'v., in the lizard Neusticurus bicarinatus. 41, group of infected thrombocytes in a lizard showing almost 100 % infection rate of these cells; 42-46, secondary schizogony, giving rise to multiple infections with developing schizonts, gametocytes and many bizarre forms. Figs. 47-52. Fallisia modestagen.nov., sp.nov., in the lizard Tropidurus torquatus hispidus. 47, 48, primary schizogony in lymphocytes; 49, lymphocyte containing a mature micro- gametocyte and a young schizont; 50, mature remate gametocyte in lymphocyte; 51,52, ruptured 'smear-cells ' liberating a macrogametocyte, a microgametocyte and merozoites from a mature schizont. Printed in GreatBritain