300 Cytologia 34

Studies of the Blood of Meloidae (Coleoptera) 1. The haemocytes of Epicauta cinerea (Forster), and a synonymy of haemocyte terminologies1

A. P. Gupta2

Department of Entomology and Economic Zoology Rutgers-The State University New Brunswick, New Jersey, U. S. A.

Received July 22, 1968

Synopsis Haemocytes of the blister beetle, Epicauta cinerea, have been de scribed. Haemocyte terminologies in many classifications of insect haemocytes have been synonymized with a view to providing a summary of different haemocyte classifications for reference.

Introduction Since Leydig's (1859) description of round and spindle-shaped cells and Magretti's (1881) and Cuenot's (1890) references to amoeboid cells in the blood of Meloe proscarabaeus L., haemocytes of several beetles have been described (Graber 1871, Lutz 1895, Hollande 1909, 1911, Barratt and Arnold 1910, Muttkowski 1924, Poisson and Patay 1939, Millara 1947, Arvy et al. 1948, Jones 1950, 1954, and McLaughlin and Allen 1965). However, none of these workers, with the exception of Hollande, has studied haemocytes of any meloid. Hollande (1911) included Lytta and Afeloe among the many Coleoptera that he studied. Beauregard (1890) has made only a passing reference to the blood cells in his extensive monograph on the blister beetles. Haemocytes of insects have been the subject of investigation for a fairly long time, and several attempts have been made to classify them. Such classifications are generally based either on the morphology and functions of the haemocytes or on their general staining and histochemical reactions. Thus, it is not unusual to find the same haemocyte or its various forms being referred to by different names, a situation which has inevitably resulted in a confusing mass of terminology. Since several physiological functions are attributed to certain haemocytes or their variant forms, it is necessary that insect haematologists follow a uniform terminology and classification in order to avoid any confusion in the interpretation and understanding of works of different authors. According to Millara (1947), Cuenot (1896) first classified haemocytes

1 Paper of the Journal Series , Agricultural Experiment Station, Rutgers-The State University New Brunswick, New Jersey. 2 Associate Professor . 1969 Studies on the Blood of Meloidae (Coleoptera) I 301 into four categories, and was later followed in this attempt by Hollande (1909 , 1911) and others. Wigglesworth (1939) summarized most of the earlier classifications, and prepared a classification which has been widely accepted . More recently, Jones (1962) revised Wigglesworth's classification with a few modifications. Jones' classification has been adopted in the present work with one exception, that is the category of prohaemocytes has not been recognized . It has been stated elsewhere (Gupta and Sutherland 1966) that prohaemocytes are earlier or young stages of plasmatocytes. The objectives of this paper are 1) to describe briefly and illustrate the haemocytes of E. cinerea , and 2) to atttempt to restore some order in haemocyte classification by synony mizing various terminologies, with a view to providing a summary of different classifications for reference.

Materials and methods

Observations were made on fixed and unfixed haemolymph, samples of which were always taken from the antennae of adult beetles of both sexes collected from wild golden-rod (Solidago sp.) plants. Examination of haemo cytes was done in hanging drops of physiological saline (NaCl, 0.9g; KCl,

0.042g; CaCl2, 0.082g; NaHCO3; 0.002g; distilled H2O, 100ml) with 2 versene. All observations were made at room temperature (28•‹-30•Ž); All photomicrographs were taken by a dark phase contrast Leitz microscope at a magnification of 400.

Observations (Figs. 1-7, 9, 11, 13, 14) Examinations of haemolymph from Epicauta, revealed the presence of plasmatocytes, granular haemocytes, spherule cells, oenocytoids, and cysto cytes. Adipohaemocytes, podocytes and vermiform cells were not observed. Plasmatocytes were predominant and were observed as round, fusiform, and lamellate cells. The cytoplasm in some of them appeared granular and the nuclei were usually centrally located. Occasionally binucleate plasmatocytes were also observed. Granular haemocytes were comparatively very rare. The nucleus in these cells was usually indistinct, due to the presence of cytoplasmic granules. Both round and fusiforms granular cells were observed. Spherule cells were found to be more numerous than the granular cells and were present either as round or fusiform cells. The spherules in the cytoplasm varied in number from two to several in various cells. Their size was also variable, sometimes the same cell showing spherules of different sizes. The nucleus in most spherule cells is usually concealed by the spherules. Mature spherule cells were observed in the process of releasing their spherules into the haemolymph. Oenocytoids, like the granular haemocytes, were compara tively very rare. They were usually fusiform in shape, although spherical forms were sometimes observed. The nucleus was generally eccentric. The 302 A. P. Gupta Cytologia 34 fi ne cytoplasmic canaliculi and granular strands, considered characteristic of oenocytoids in Galleria and other insects, were not observed. The cystocytes are usually hyaline cells with characteristic cartwheel-like nuclei. They are usually fewer in number than other haemocytes, although several of them are

Figs. 1-7. 1 and 2, plasmatocyte (Epicauta cinerea). 3 and 4, granular haemocyte (E . cinerea). 5 and 6, spherule cell (E. cinerea). 7, oenocytoid (E. cinerea). 1969 Studies on the Blood of Meloidae (Coleoptera) I 303 observed in a coagulated sample of haemolymph. In addition to the above inentioned haemocytes, clusters of spherules were also observed in the haemo lymph. In several samples of unfixed haemolymph, coagulation spots were formed, and these provided material for study of coagulation patterns and

Figs. 8-11. 8, podocyte (Periplaneta americana nymph). 9, cystocyte (E. cinerea). 10, adipohaemocyte (Galleria mellonella). 11, cluster of spherules; notice that the spherules look nucleated (E. cinerea). 304 A. P. Gupta Cytologia 34

Figs. 12-14. 12, vermiform cell (P. americana). 13 and 14, coagulation patterns (E. cinerea). 1969 Studies on the Blood of Meloidae (Coleoptera) I 305

the role of haemocytes, if any, in the process of coagulation . Examination of several coagulation spots showed only cystocytes . Whether cystocytes actually cause coagulation or are the result of coagulation is difficult to say . It should be noted here that in samples in which a differential count would show very few cystocytes, induced coagulation would reveal several of these cells. Probably during such induced coagulation, certain fragile plasmatocytes are unable to withstand the mechanical stress-and perhaps some physiological stress in vivo-and suddenly release their cytoplasmic contents, which form the peculiar cytoplasmic strands, so characteristic of the coagulation pattern . These granular cytoplasmic strands ramify randomly and enclose several cystocytes. No granular islands around the cystocytes were observed, although cytoplasmic veils enveloping all of the cytoplasmic strands, and the cystocytes were visible. Occasionally, coagulation spots without cytoplasmic strands were also observed.

Synonymy of haemocyte terminologies The terminologies synonymized in Table 1 are variously based on stained or unstained fresh or fixed haemolymph. Some are based on histochemical studies on the haemocytes. Some descriptions included haemocytes of only immature stages, some of adults, and some of both stages. Homologies have been established on the basis of description, line drawings, and photomicro graphs of original authors. Cells which could not be homologized have been included in the category of "others." In some cases cells included in this category may not necessarily be haemocytes. Generous use of footnotes has been made to include pertinent information on the original classifications. Except in a few cases, only Anglicized versions of original terms have been included in the table. Terms in parenthesis were not used by the original authors, but were adopted by the present writer after scrutinizing original descriptions and figures, and represent only approximations to the category they have been assigned to in the table. The identity of term "with?" is uncertain. Haemocytes categorized as phagocytes and amoebocytes by previous authors have been assigned to the category of plasmatocytes, although they could also be included with granular haemocytes, spherule cells, and adipo haemocytes, inasmuch as these latter three forms are also phagocytic in nature. No attempt has been made to include the current generic and specific names of the species by many of the earlier authors. Where no specific name of the insect was given by the original author only common familial or ordinal names have been included. Ordinal designations in the table have been included for all the species, whether these taxonomic designations were given by the original authors or not. The name of the author of every species has been omitted. Taxonomic designations were checked from Cresson (1887), Kirby (1892), Heyden et al. (1906), Blatchley (1920), Leng (1920), Brues et al. (1954), Harz (1957), Borror and Delong (1960), and Imms (1960). All Table 1.

(Continued) Table 1. (continued)

1 Graber 1871. Although he has attempted to arrange the species he studied under four categories, several of the insects in each category show more than one form of haemocyte. (Continued) Table 1. (Continued)

(Continued) Table 1. (Continued)

2 Magretti 1881. Some of his figures indicate podocytes; he also figured coagulation pattern in Meloe. 3 Lutz 1895. Describes cells only by their shape, and does not use any definite term. (Continued) Table 1. (Continued)

(Continued) Table 1. (Continued)

* Kollman 1908. According to him granular cells are absent in Coleoptera, Lepidoptera, Hymenoptera and Diptera. (Continued) Table 1. (Continued)

5 Hollande 1909. Although he did not find granular haemocytes and oenocytoids in Coleoptera, he recognized their presence and also that of "adipoleucocytes" in other insects. (Continued) Table 1. (Continued)

6 Poyarkoff 1910. He considered oenocytoids homologous to the lymphocytes of Hollande (1911). (Continued) Table 1. (Continued)

(Continued) Table 1. (Continued)

7 Tillyard 1917. From his description, it appears that his 'miocytes' are plasmatocytes, although he could have mistaken muscle cells for haemo cytes. (Continued) Table 1. (Continued)

8 Muttkowski 1924. Presents only line drawings. His stage III might contain granular haemocytes, spherule cells and adipohaemocytes. (Continued) Table 1. (Continued)

9 Poisson 1924. His leucocytoid and oenocyte correspond to oenocytoid of Hollande (1911). (Continued) Table 1. (Continued)

(Continued) Table 1. (Continued)

10 Paillot and Noel 1928. They considered proleucocytes and phagocytes of Hollande (1909, 1911) as macronucleocytes. According to them these cells are not phagocytic but micronucleocytes are. From their figures, it appears that the so-called prohaemocytes and the macronucleo cytes are plasmatocytes. (Continued) Table 1. (Continued)

(Continued) Table 1. (Continued)

(Continued) Table 1. (Continued)

11 Rooseboom 1937. Her granular haemocytes look more like spherule cells. (Continued) Table 1. (Continued)

(Continued) Table 1. (Continued)

(Continued) Table 1. (Continued)

12 Millara 1947. She included macro- and micronucleocytes in phagocytes; according to her, young leucocytes (recognized by Hufnagel, Poisson, and Hamilton) are intermediate stages between proleucocytes and phagocytes; she does not recognize spherule cell as a distinct type and thinks these are derived from granular haemocytes. (Continued) Table 1. (Continued)

(Continued) Table 1. (Continued)

13 Arvy et al . 1948. Their leucoblast corresponds to the so-called proleucocytes of other authors. 14 Arvy et at . 1949. According to them, their " Leucocytes a cytoplasm basophile" correspond to the so-called lymphocytes of some authors. (Continued) Table 1. (Continued)

15 Akesson 1954 . His type II seems to be a plasmatocyte. (Continued) Table 1. (Continued)

(Continued) Table 1. (Continued)

16 Wigglesworth 1956 . According to him phagocytic amoebocytes include plasmatocytes, podocytes and spheroidocytes and large granular and agranular cells. His amoebocytes seem to include spherule cells as well. (Continued) Table 1. (Continued)

17 Arnold 1959. Recognizes only two types, but his figures 12 and 16 indicate granular hemocytes and figure 19 a vermiform cell . 18 Hrdy 1960. Although he synonymizes his rhegmatocyte with Gregoire's coagulocyte (=cystocyte), the photomicrographs suggest that it is perhaps a young plasmatocyte, and his spheroidocyte (Fig. 11, Plate II) looks like a granular haemocyte. (Continued) Table 1. (Continued)

19 Nittono 1960. Considered imaginal spherule cells as a separate category, and has not homologized it with any other haemocyte of other authors. (Continued) Table 1. (Continued)

2) Jones 1965. Considers Wigglesworth's (1956) oenocytoid in R. prolixus as a granular cell. Wigglesworth (1966, personal communi cation), however, does not agree with Jones' designation of granular cells and oenocytoids. Wigglesworth also believes that the adipocytes in Rhodnius are in fact detached fat-body cells. (Continued) Table 1. (Continued)

21 Wille and Vecchi 1966. Compared Picnonuclocyte with Yeager's (1945) Eruptive cell (=spherule cells), and they have not mentioned spherule cells. It seems that their Prochromatinocytes and Chromatinocytes are various stages of spherule cells . Their Oenocytoids seems to be doubtful, since they look like degenerate Plasmatocytes. 1969 Studies on the Blood of Meloidae (Coleoptera) I 335 classifications in the table are arranged chronologically. Table 1 does not necessarily include all haemocyte classifications that have been proposed to date. Since podocytes, vermiform cells and adipohaemocytes were not observed in Epicauta, photomicrographs of these cells were taken from Periplaneta americana (L.) and Galleria mellonella L. for the purpose of illustration (Figs. 8, 10 and 12).

Discussion As far as is known only Hollande (1911) has attempted to distinguish various forms of haemocytes in any blister beetle. It appears from the literature that no single species of beetle possesses all seven types of haemo cytes included in Table 1. For example, of those authors who have recognized various types of haemocytes in beetles, Hollande (1909, 1911), Poisson and Patay (1939), Millara (1947), and Arvy et al. (1948) have not reported adipo haemocytes, podocytes, vermiform cells, and cystocytes in the beetles they studied. The present writer also did not observe adipohaemocytes, podocytes, and vermiform cells in E. cinerea. Jones (1950), however, has reported adipohaemocytes (=spheroidocytes) and vermiform cells in Tenebrio molitor L. It has been stated earlier that prohaemocytes are not recognized as a distinct type by Gupta and Sutherland (1966). Earlier, Jones (1956) considered them as miniature plasmatocytes, and Clark and Harvey (1965) have supported Jones' observation. Granular haemocytes have been reported in a few beetles (Hollande 1911, Millara 1947 and Arvy et al. 1948). Jones (1950, 1954) and McLaughlin and Allen (1965) did not observe granular cell in Tenebrio molitor and Anthonomus grandis Boheman, respectively. In E. cinerea, granular haemocytes are present but are rarely observed. Hollande (1909) for the first time demonstrated the presence of spherule cells in several beetles. These haemocytes, however, were not observed by Barratt and Arnold (1910), Muttkowski (1924), Poisson and Patay (1939), Arvy et al. (1948), and Jones (1950, 1954) in various beetles that they studied. Only Millara (1947) and McLaughlin and Allen (1965) have reported spherule cells in their studies. Spherule cells in Epicauta are comparatively more abundant than granular haemocytes. Oenocytoids were first observed in several beetles by Hollande (1911). Subsequently, these cells were reported only by Poisson and Patay (1939), Millara (1947), and Jones (1950, 1954). In E. cinerea oenocytoids are present but are rarer than granular haemocytes. Jones (1950, 1954) and McLaughlin and Allen (1965) demonstrated cystocytes in T. molitor and A. grandis, respectively. These haemocytes were occasionally observed in E. cinerea. Clusters of spherules reported in the present study have also been observed by the present writer in several species of cockroaches, wax moth, and mealworm. Gupta and Sutherland (1967) have suggested that these clusters of spherules are probably precursors of new cells. 336 A. P. Gupta Cytologia 34

Coagulation patterns have been described by Gregoire (1951, 1955, 1957 and 1959) in a few blister beetles: Meloe proscarabaeus L., M. violaceus Marsh, several species of Mylabris (Zonabris) and Epicauta grammica (Waldheim). Based on studies of coagulation processes in several insects, Gregoire (1955, 1957) proposed four coagulation patterns. According to him, Meloe, Zonabris and Epicauta all show predominantly Pattern I, in which islands of coagulation form around the cystocytes. Zonabris and Epicauta, however, also show Pattern III, which is a combination of patterns I and II. In the latter pattern, the cystocytes form several granular cytoplasmic strands, which in certain cases are embedded in glassy veils of plasmal origin. In Epicauta cinerea, the coagulation pattern does not fall into either pattern I or III, as described by Gregoire in the blister beetles he studied. In this species, the coagulation pattern more or less corresponds to Gregoire's type II, Magretti's (1881) Figure 2 also vaguely suggests pattern II in Meloe. On the basis of his study of coagulation processes in several species of insects, Gregoire (1957) considered pattern I as the most generalized and primitive type and pattern II as belonging to relatively recent groups of insects. Within the family Meloidae, it is difficult to say which of the two patterns is the more generalized type, since only three genera have so far been studied. It should be noted here that on the basis of the internal anatomy, the present writer (Gupta 1965) has shown that Meloe and Epicauta are more closely related to each other than either one of these genera is to Mylabris. It seems that perhaps pattern III is the basic type at least in the subfamily Meloinae, to which all these genera belong (Kaszab 1959, Gupta 1965), and from this basic pattern patterns I and II secondarily evolved. A detailed study of the coagulation processes in several genera of the Meloidae is needed. As far as is known, no comprehensive synonymy of haemocyte termi nologies is available. Munson (1953), Nittono (1960), and Choudhuri (1964) have presented brief synonymies in a few classifications, the last two authors dealing only with those concerning some Lepidoptera and Orthoptera, re spectively. Cuenot (1890) was perhaps the first to use a specific terminology (amoebocyte) for blood cells in insects. Most authors, however, both before and after Cuenot, continued to describe cells by their shapes (Table 1). Graber (1871) probably made the first attempt to arrange various insects into four groups on the basis of haemocytes, although several of the insects in each of his groups show more than one form of haemocyte. Cuenot's (1890) term "amoebocyte" has been adopted b y several authors (Mercier 1906, Tillyard 1917, Muttkowski 1924, Hamilton 1931, Poisson and Patay 1939, and Wiggles worth 1956). Of these authors Hamilton and Poisson and Patay have used the terms leucocyte and phagocyte, respectively, as synonymous to amoebo cytes. It seems that Kollman (1908) and Metalnikov (1908) first adopted the term leucocyte, and the following year Hollande (1909) used the term phagocyte . He also introduced the term lymphocyte at about this time. It should be 1969 Studies on the Blood of Meloidae (Coleoptera) I 337 noted here that Hufnagel (1918), Metalnikow and Gaschen (1922) , Glaser (1927) and Metalnikoff (1927) considered leucocyte and phagocyte as being synonymous. Several authors (Hollande 1909, 1911, Poyarkoff 1910, Hufnagel 1918, Ishimori 1925, Glaser 1927, Metalnikoff 1927 , Mathur and Soni 1937, Dennell 1947, Millara 1947, Akesson 1954 , Ogel 1955, and Choudhuri 1964) have variously recognized the categories of proleucocytes, granular cells, spherule cells, adipohaemocytes, and oenocytoids , and have also retained the category of phagocytes in their respective classifications . Recognition of the category of phagocytes with those of prohaemocytes, granular cells, spherule cell and adipohaemocyte in the same classification seems to be inconsistent in view of the fact that phagocytosis has been reported in prohaemocytes (Yeager 1945), granular cells (Yeager 1945 and Jones 1956), spherule cells (Kollman 1908, Cameron 1934 and Akesson 1954), and in adipohaemocytes (Zeller 1938 and Yeager 1945). Of the three terms (leucocyte, phagocyte, and lymphocyte) the first two are still used by some workers while the latter seems to have fallen into disuse. The term which has survived to date with little or no change since its adoption by Hollande in 1911, is proleucocyte. Arnold (1952) used the term prohaemocyte, which is commonly used by American authors, the European workers still preferring the term proleucocyte. Paillot (1919) introduced two new terms, micro and macronucleocyte on the basis of the size of the nucleus and the amount of cytoplasm. The macronucleocytes are homologous to the so-called proleucocytes or prohaemocytes. and the micronucleocytes to the leucocytes, the phagocytes, and the lymphocytes. It is interesting to note that Hollande did not recognize Paillot's terminology until 1928 (Hollande and Aghar). Other workers who adopted micro and macronucleocytes in their classifications are: Aghar (1928), Bogojavlensky (1932), Rooseboom (1937), Lepesme (1938), Wigglesworth (1939), Chauvin (1956), and Tuzet and Manier (1959). Yeager (1945) introduced several new terminologies of insect haemocytes, the most generally accepted of which are plasmatocyte, podocyte, vermiform cell, and the cystocyte, the latter term being originally meant for granular cells. Plasmatocyte is equivalent to what other workers call leucocyte, phagocyte, or lymphocyte. Podocyte probably included star-shaped, amoeba like cells of Graber (1871) and radiate cells of Lutz (1895). Jones (1950) for the first time applied the term cystocyte to those haemocytes which cause coagulation, and which have been referred to as coagulocytes by Gregoire (1951). Jones (1846) first established the category of granular cells, and later Cuenot (1896) mentioned amoebocytes with finely granular cytoplasm. Hollande (1909) should undoubtedly be credited with the introduction of the term spherule cell, which has been referred to only as spherule by a few authors (Poyarkoff 1910, Patton 1963 and Whitten 1964). Hollande (1911) also intro duced the term adipoleucocyte, although Kollman (1908) had earlier used the 338 A. P. Gupta Cytologia 34 term adipo-spherule cell in his study of the haemocytes of some invertebrates. Wigglesworth (1965) has called these cells adipocytes. No two terms have caused so much confusion as oenocytes and oenocytoids. Oenocytes differ from oenocytoids in that they are ectodermal in orgin, are usually segmentally arranged, are generally yellow in color, and are not haemocytes, although in Gerris they supposedly give rise to oenocytoids (Wigglesworth 1965). Hollande (1911) introduced the term oenocytoid and later (1920) gave a detailed description of these cells. In order to avoid any confusion between the terms oenocytes (proposed by Wielowiejski 1886) and oenocytoids, Hollande (1914) proposed to replace the term oenocyte by "cerodecytes ." It is not unlikely that several earlier authors mistook oenocytes for oenocytoids. Even after Hollande's (1920) detailed description of oenocy toids, several authors (Metalnikow and Gaschen 1922, Muller 1925, Tateiwa 1928, Metalnikov and Chorine 1929, Bogojavlensky 1932 and Cameron 1934) have used the term oenocvte instead of oenocvtoid in their respective works. In addition to the above-mentioned haemocyte terminologies, several authors have, from time to time, proposed terms, many or all of which have not been generally accepted. For example, haemocytoblast of Bogojavlensky (1932); leucoblast of Arvy and associates (1944, 1948), and proleucocytoid and prohaemocytoid of Yeager (1945) and Jones (1950), respectively. Yeager also introduced the term nematocyte and proposed the term spheroidocyte for adipohaemocyte. Rizki in his works used the terms lamellocyte (1962), and crystal cell (1962), the latter has also been adopted by Whitten (1964) and is probably an oenocytoid. Terms such as seleniform cells (Poyarkoff 1910), miocytes (Tillyard 1917), splanchnocytes (Muttkowski 1924), teratocytes (Hollande 1920), pycnonucleocyte (Morganthaler 1953, Wille and Vecchi 1966), nucleocyte, and rhegmatocyte (Hrdy 1957) are rarely encountered in the literature. Finally, Jones (1965) introduced the term granulocytophagous cell in his work on Rhodnius prolixus Stal, and Scharrer (1965) called Ritter's (1965) "anucleate crescent body" a crescent cell in the cockroach, Grom phadorhina portentosa Schaum. Examination of the many species listed in Table 1 reveals that, as far as is known, haemocytes in several orders of insects have not been studied. For example, among the Apterygote orders, only Thysanura have been studied. Of the two Paleopterous orders Odonata and Emphemeroptera, the latter have apparently not yet been studied. Among the Orthopteroid group, Isoptera and Embioptera still await studies. In the Hemipteroid complex, no account of haemocytes is available in Zoraptera, Phthiraptera, Corrodontia, and Thysanoptera, and finally, in the Neuropteroid group haemocytes have not yet been studied in Raphidoidea, Megaloptera, Neuroptera, Mecoptera, and Siphonaptera. In terms of number of species studied in various orders, Orthoptera and Coleoptera appear to be the most extensively studied groups. Next to these are perhaps Lepidoptera, Hymenoptera, and the Diptera, in 1969 Studies on the Blood of Meloidae (Coleoptera) I 339 that order. In addition to the Heteroptera , Homoptera, and Odonata, of which only a few species have so far been studied , Dermaptera, Plecoptera, Trichoptera, and Thysanura are the most poorly studied groups . Table 1 reveals that, with the exception of plasmatocytes , and perhaps also the granular haemocytes, all other types of haemocytes are not present in all insect orders. For example, no spherule cells , adipohaemocytes, oenocytoids, podocytes, vermiform cells, and cystocytes have been reported in Thysanura, Odonata, Dermaptera, Plecoptera and Trichoptera. The present writer, however, found (1967) spherule cell and cystocytes in Thysanura and Dermaptera. Granular haemocytes, spherule cells, and adipohaemocytes are lacking in Hymenoptera; and Heteroptera and Homoptera do not seem to possess spherule cells, podocytes, vermiform cells, and cystocytes. Only in Orthoptera, Coleoptera, Lepidoptera, and Diptera have all types of haemocytes been reported. It must be emphasized, however, that several of the species listed in Table 1 were studied at a time when very little was known about insect haemocytes, and different authors used different terminologies to suit their descriptions. It is obvious from the foregoing that a detailed study of haemocytes in most insect orders is needed. Such a study would place the presently recognized classification on a firmer ground. It might also reveal some haemocyte types, which have not yet been reported. Furthermore, it would enable us to speculate on the probable evolutionary stages of various haemocyte types and their physiological roles. The present writer, realizing the importance of a comprehensive account of hemocytes in Insecta, hopes to study hemocytes in various insect orders and will report the results later.

Summary

Epicauta cinerea possesses plasmatocytes, granular haemocytes, spherule cells, oenocytoids, and cystocytes. Adipohemocytes, podocytes and vermiform cells are absent in the blood of this beetle. Haemocyte terminologies have been synonymized, using several classifi cations by various authors. This study revealed that haemocytes have not yet been studied in Protura, Diplura, Collembola, Isoptera, Embioptera, Zoraptera, Phthiraptera, Corrodontia, Thysanoptera, Raphidoidea, Megaloptera, Neuroptera, Mecoptera, and Siphonaptera. In terms of number of species studied, Orthoptera and Coleoptera are the most extensively studied groups, while Dermaptera, Plecoptera, Trichoptera and Thysanura are the most poorly studied orders. Of all the orders, only Orthoptera, Coleoptera, Lepidoptera, and Diptera, have all the 7 or 8 types of haemocytes.

Acknowledgments

I am thankful to Professor Sir Vincent B. Wigglesworth, Cambridge, England, for reading the initial manuscript, and to Professor John B. Schmitt 340 A. P. Gupta Cytologia 34 of our department for providing speciment of Anisolabis maritima (Gene). The secretarial assistance of Mrs. Florence B. Stanik is sincerely appreciated.

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