Morphology of the male reproductive system and sperm of Leptoglossus zonatus (Dallas, 1852) (Heteroptera: Coreidae).

Taxonomic data on Coreidae have been fragmented over time and need to be revised. Likewise, data related to the development of germ cells and the features of the male reproductive system, including sperm, will contribute to understanding the biological mechanisms of reproduction and the systematics of its representatives. Aiming to provide these data, we describe the morphology of the male reproductive system and spermatozoa of Leptoglossus zonatus using light and transmission electron microscopies, respectively. Each of the two testes is surrounded by a bright red-pigmented sheath and formed by seven follicles arranged side by side. The two vasa deferentia are filled with individualized sperm, especially in their final portion, which is dilated and curved. After dilation, the vasa deferentia receive the ducts of the accessory glands of mesodermal origin. The other unpaired accessory gland is of ectodermal origin and opens into the ejaculatory duct. Both glandular types are densely coiled and have lumens filled with secreted material. Testicular follicles contain cysts with germ cells at different stages of spermatogenesis, indicating continuous production of gametes throughout adult life. Mature sperm measure around 310 µm long, with a nucleus of 36 µm and a flagellum formed only by an axoneme of 9 + 9 + 2 microtubules and two symmetrical mitochondrial derivatives. Like the sperm of other Heteroptera, the acrosome has a single structure (without perforatorium), there are no accessory bodies in the flagella, and the mitochondrial derivatives are connected to the axonemes, supporting the synapomorphic condition of these characteristics for this suborder of bedbugs. RESEARCH HIGHLIGHTS: The Leptoglossus zonatus sperm are slender and long, about 310 µm in length, and a nucleus 36 µm long. Spermatogenesis occurs throughout adult life and equally in the seven testicular follicles. The centriole adjunct in L. zonatus sperm does not give rise to accessory bodies. The ectodermal gland produces a filamentous secretion, whereas in the ectodermal sac, the secretion is globular.

Sperm morphology of Tingidae Laporte, 1833 (Miroidea: Cimicomorpha).

Here, we describe for the first time the sperm morphology of Tingidae (Heteroptera). They are small insects presenting lacy patterns on their pronotum and hemielytra and are exclusively phytophagous, with many economically important species. We studied five species of the tribe Tingini (Tinginae): Teleonemia scrupulosa, Vatiga illudens, Gargaphia lunulata, Leptopharsa sp., and Corythucha arcuata. Their spermiogenesis process is similar to other Heteroptera, with some differences in the formation of the centriole adjunct. This structure extends in the anteroposterior spermatid axis, flanking the nucleus, possibly contributing to nucleus remodeling and sperm elongation. The mature sperm of Tingidae is also similar to that of other Heteroptera, with features that corroborate the group's monophyly. Our data support previous results for their sister family, Miridae, which exhibits some characteristics exclusive to this taxon, not present in Tingidae or other Heteroptera. They also support the sister relationship of the genera Gargaphia and Leptopharsa and suggest closer relationship between Vatiga and Corythucha. Overall, this study sheds light on the sperm ultrastructure of Tingidae and provides information for understanding the evolution and diversity of Heteroptera. RESEARCH HIGHLIGHTS: The spermiogenesis process and mature sperm are similar to other Heteroptera The centriole adjunct is derived from a strip of a pericentriolar material extending from the centriole Tingidae and Miridae are distinguishable using sperm morphology.

Morphology of the male reproductive system and spermatozoa of Smicridea (Rhyacophylax) iguazu Flint, 1983 (Trichoptera, Hydropsychidae).

The Trichoptera, holometabolous aquatic insects found worldwide except in Antarctica, exhibit a unique feature in their sperm, which are solely nucleated (eupyrene). Current knowledge on Trichoptera sperm is limited to Old World species. To enhance our understanding of their reproductive biology and contribute to systematic discussions, we describe the male reproductive system and spermatozoa of Smicridea (Rhyacophylax) iguazu Flint, 1983 (Hydropsychidae). This species lacks seminal vesicles, possesses piriform to oval-shaped testes with spermatozoa grouped in apical bundles and dense filamentous material filling other areas. The vasa deferentia are long and a pair of elongated accessory glands displays distinct proximal and distal regions. The relatively short (∼40 µm) spermatozoa are nucleated, aflagellated, and immobile. Further research could explore variations and assess the taxonomic utility of these features for genus identification within Hydropsychidae.

Morphology of the male reproductive system and spermatozoa of Mahanarva fimbriolata (Stål, 1854) (Hemiptera: Cercopidae) with evidence of a spermatostyle in the sperm conjugation.

Changes in harvesting methods in the early 1990s led to Mahanarva fimbriolata becoming the most prevalent pest in sugarcane plantations. Encouraged by the scarcity of research on the reproductive system and spermatology of Auchenorrhyncha, we provide a detailed description of the male reproductive system and spermatozoa of this cercopid. This will not only aid in taxonomic purposes but also help in understanding the reproductive biology of the species. The male reproductive system, spermatozoa, and sperm bundles of M. fimbriolata were investigated using light microscopy. Our results reveal a reproductive system consisting of a pair of testes, each with 36 follicles, two vasa deferentia, two well-developed seminal vesicles, a single pair of accessory glands, and an ejaculatory duct. Here, we also describe and explore the particularities of the formation of secondary sperm bundles, where cells interestingly bind only to the apical region of the spermatostyle. We also propose some possible benefits of this unusual organization for the reproductive success of the species.

Morphology of the Female and Male Reproductive Tracts and More Data on the Spermatostyle in the Brazilian Gyretes sp. (Coleoptera, Adephaga, Gyrinidae).

We investigated the male and female reproductive tracts of Gyretes sp. with light and transmission electron microscopies. The male has a pair of testes with a single coiled follicle, followed by short efferent ducts, which have a similar shape and diameter to the testes. Long ducts (epididymides) with differential epithelium open in a pair of long vasa deferentia that lead to the accessory glands. Glycoprotein secretions from the vas deferens epithelium constitute the spermatostyle for spermatozoa aggregation. The female has numerous ovarioles per ovary, a coiled fertilization duct, an accessory gland, and an elongated vagina. Spermatozoa are stored as unaggregated cells in the fertilization duct. In Gyrinidae, the testes and accessory glands show diverse shapes, and the female sperm storage organs vary in shape, size, and type and may play a role in the interaction with sperm aggregates. Testes with a single follicle and vasa deferentia opening in the accessory glands of Gyretes sp. are features shared with other Gyrinidae and other Adephaga. We proposed adding this latter trait to characterize this suborder of beetles. The morphology of the reproductive organs in both sexes contributes to comparative analyses and knowledge of the reproductive biology of Gyretes and may provide additional features for systematics.

Sperm ultrastructure of Pycnoderes incurvus (Hemiptera: Miridae).

The sperm morphology can provide helpful information about sexual selection, phylogeny, and the evolutionary history of a given animal group. However, there is limited or no knowledge of many taxa, especially those belonging to insects, a vast and highly diverse group. An example is the Miridae, or plant bugs, which belong to the infraorder Cimicomorpha (Heteroptera), where only three out of 17 families have published data on their sperm morphology. Here we described the Miridae sperm structure by analyzing Pycnoderes incurvus sperm under light and transmission electron microscopy. In this species, the spermatozoa were as long and slender as those of most insects. However, the anterior-most region was twisted, a characteristic first reported for Heteroptera. The acrosome was coated with electron-dense material, most likely extra-acrosomal. The centriole adjunct was a notably long, cylindrical and compact structure connecting the nucleus to the flagellar elements, with just clove-like electron-lucent points in cross-section, also features unique to Miridae so far. The flagella exhibited an axoneme of 9 + 9 + 2 microtubules and two symmetrical mitochondrial derivatives. The latter two partially embrace the axoneme, and each exhibits two paracrystalline areas and a bridge connecting it to the axoneme; these are considered Heteroptera synapomorphies that support their monophyly. RESEARCH HIGHLIGHTS: The P. incurvus sperm showed a twisted acrosome, the first reported for Heteroptera. The centriolar adjunct is a sole structure linking the nucleus and flagellum. The flagella presented the synapomorphies supporting Heteroptera monophyly.

First Record of Zaprionus tuberculatus Malloch, 1932 (Diptera: Drosophilidae) in Minas Gerais, Brazil

ABSTRACT The Afrotropical Zaprionus Coquillett, 1902 (Diptera: Drosophilidae) has gained visibility since the 1990s due to the geographic expansion of Zaprionus indianus Gupta, 1970. More recently, a second species, Zaprionus tuberculatus Malloch, 1932, invaded regions outside its original African regions, causing economic concerns, particularly in Europe. In 2021, Z. tuberculatus was captured for the first time in the Americas, specifically in urban parks, and preserved fragments of the Brazilian Cerrado, causing concerns about the competition with native drosophilids. Here we report the occurrence of Z. tuberculatus 900 km from its first record in urban and rural areas of Viçosa, MG, but not in forest fragments. Considering the great capacity for dispersion and the potential of Z. tuberculatus to compete with native drosophilids species, as well as the potential harm to fruit production when co-occurring with Drosophila suzukii (Matsumura, 1931), further studies would be necessary to monitor this invasion and create mechanisms to control it.

New Findings on the Sperm Structure of Tenebrionoidea (Insecta, Coleoptera).

The sperm ultrastructure of a few representative species of Tenebrionoidea was studied. Two species belong to the Mordellidae (Mordellistena brevicauda and Hoshihananomia sp.), one species to Oedemeridae (Oedemera nobilis), and one species to Tenebrionidae (Accanthopus velikensis). It is confirmed that Mordellidae are characterized by the lowest number of spermatozoa per cyst (up to 64), a number shared with Ripiphoridae. In contrast, in the two other families, up to 512 spermatozoa per cyst are observed, the same number present, for example, in Tenebrionidae. Also, as in the other more derived families of tenebrionoids studied so far, during spermatogenesis in O. nobilis and A. velikensis, sperm nuclei are regularly distributed in two sets at opposite poles of the cysts. On the contrary, the Mordellidae species do not exhibit this peculiar process. However, during spermiogenesis, the bundles of sperm bend to form a loop in their median region, quite evident in the Hoshihananomia sp., characterized by long sperm. This process, which also occurs in Ripiphoridae, probably enables individuals to produce long sperm without an increase in testicular volume. The sperm looping could be a consequence of the asynchronous growth between cyst size and sperm length. The sperm ultrastructure of the Mordellidae species reveals that they can be differentiated from other Tenebrionoidea based on the shape and size of some sperm components, such as the accessory bodies and the mitochondrial derivatives. They also show an uncommon stiff and immotile posterior flagellar region provided with only accessory tubules. These results contribute to a better knowledge of the phylogenetic relationship of the basal families of the large group of Tenebrionoidea.

The sperm ultrastructure of members of basal Tenebrionoidea (Coleoptera).

The sperm ultrastructure of some beetles of Tenebrionoidea was studied with particular attention to those of the Ripiphoridae, Mordellidae, and Meloidae. These three groups are often thought to form a clade, which is the sister group of the remaining Tenebrionoidea. The testes of the two former families have thinner but longer spermatic cysts containing fewer and longer sperm. Within each cyst all sperm cells have the same orientation, but cross sections showed that the orientation of the axonemes alternate between adjacent cysts, possibly due to the cysts bending on themselves. In both families the sperm has a bilayered acrosome and the flagellum, which shows mitochondrial derivatives starting laterally to the nuclear base, has a typical 9 + 9+2 axoneme with accessory tubules provided with 16 protofilaments in their wall, and well-structured triangular shaped accessory bodies. In Mordellistena sp (Mordellidae) sperm, both mitochondrial derivatives and accessory bodies are somewhat asymmetrical. Moreover, the flagellum shows a very thin and long tail end provided with only accessory tubules. Meloidae species have testes with thicker sperm cysts containing numerous shorter sperm. Within the individual cysts the sperm flagella exhibit an alternating orientation of their axonemes as consequence of a peculiar spermatogenetic process. The flagellar structure is similar to that of the above-mentioned species, but the accessory bodies are not well defined and constituted by fuzzy material. In Mylabris hieracii (Meloidae) sperm, the acrosome is flat with a conspicuous perforatorium and its nucleus has a peculiar quadrangular section. Berberomeloe majalis sperm has a large acrosome with an unusual pentagonal perforatorium. The centriolar structure of Mylabris variabilis shows a complex of dense radial links connecting the microtubular structures to the plasma membrane. These results suggest that Ripiphoridae have a closer relationship with Mordellidae than with Meloidae. These findings are in agreement with results obtained with molecular data.

The sperm ultrastructure of Pytho depressus (Linnaeus, 1767) (Coleoptera, Pythidae).

The sperm ultrastructure of Pytho depressus (Pythidae) is described in this study. The sperm are short cells, about 85-90 µm long, with an acrosome consisting of three layers, a cylindrical nucleus, which at its base has the initial region of two mitochondrial derivatives. The flagellum has two well-developed triangular accessory bodies, and a 9 + 9+2 axonemal pattern with accessory tubules provided with 16 protofilaments in their wall. The structure and shape of the accessory bodies are diagnostic characters within the superfamily. The sperm morphology of P. depressus can be easily distinguished from those of Ripiphoridae, Meloidae and Tenebrionidae. The P. depressus sperm are organized in cysts as in other species of the group but the sperm are not well aligned and show an antiparallel orientation, a feature also observed in other tenebrionids. The phylogenetic implications of the observed sperm features are discussed in the context of comparative sperm ultrastructure of other insect species.

The sperm ultrastructure of the click beetles (Elateridae) and related groups (Buprestidae and Lampyridae).

In the present study, we describe the sperm morphology of 11 species of Elateriformia (9 elaterids, 1 lampyrid and 1 buprestid) using transmission electron microscopy. All species exhibited sperm that is not usually observed in insects in general. The most highlighted features are the displacement of the nucleus running parallel to the flagellar components, hitherto observed only in coccinellid and carabid beetles, and the presence of thin and dense structures along the nucleus, probably derived from the centriole adjunct, a feature that is so far exclusive to these insects. The other structures are a typical axoneme for insects with 9 + 9 + 2 microtubules, in a position diametrically opposite relative to the nucleus, two slender, symmetrical mitochondrial derivatives and a pair of discrete accessory bodies. This arrangement provides a bilaterally symmetrical flagellum, which favourably influences sperm hydrodynamics, as will be discussed. The occurrence of this unusual structural arrangement in the sperm of species from superfamilies that are phylogenetically as distant as Elateroidea and Buprestoidea support the monophyly of the infraorder Elateriformia, as proposed by some previous molecular studies.

Sperm ultrastructure of three species of ladybirds (Coleoptera, Coccinellidae).

The sperm of three coccinellid species belonging to the subfamilies Chilocorinae, Coccinellinae and Epilachninae were studied under light and transmission electron microscopy. The basic sperm structure of these ladybirds is common to that of the other previously studied species, especially the acrosome in front of the basal body and not the nucleus, with this latter running parallel with the flagellar components. In the Chilocorinae Platynaspis luteorubra (Platynaspidini) the sperm are of the type 1, as in Scymnini and Coccinellini, since they exhibited a cylindrical basal body with 9 + 0 and then 9 + 2, microtubules continuing further in an initial flagellar portion with the only axoneme devoid of accessory structures. The sperm exhibits a thin nucleus and mitochondrial derivatives. Such uniformity of sperm ultrastructure could be indicative of the occurrence of a close relationship between Platynaspidini and Scymnini as also proposed in the previous studies. Conversely, they differ markedly from the sperm of type 3 observed in the Chilocorini Exochomus quadripustulatus. In the Coccinellini Propylea quatuordecimpunctata the sperm are also of the type 1, but they can be easily differentiated from those of the other Coccinellinae studied so far, because of their very short acrosome without the posterior extension, the relatively thicker mitochondrial derivatives and the cylindrical nucleus. Epilachna clandestina sperm resemble those of E. quadripustulatus but differ from them, because they exhibit an elliptical nucleus which is anteriorly very thin, and the asymmetrical mitochondrial derivatives in the anterior extremity, with the greater one starting at the same level of the basal body, rather than at the nucleus level, as it occurs in E. quadripustulatus. Because of the differences observed in the sperm ultrastructure we propose a new sperm type (Type 4) for E. clandestina. This study on ladybird spermatozoon ultrastructure clearly indicates that the current classifications of coccinellids do not reflect the natural history of this well-known insect family.

Sperm morphology of Elasmus polistis Burks, 1971 (Hymenoptera: Chalcidoidea: Eulophidae).

The sperm morphology of the parasitoid Elasmus polistis (Eulophidae) has been investigated with light and transmission electron microscopy. The sperm were filiform and spiraled, with 165.6 (± 4.6) µm in length, and showed a distinctive head, formed by a one-layered small acrosome and a nucleus, and a flagellar region. An extracellular sheath from which many long filaments radiated out covered the acrosome and part of the nucleus. The spiral nucleus, with 24.1 (± 1.3) µm in length, was filled with homogeneously compact chromatin. In the nucleus-flagellum transition, the centriole adjunct extended posteriorly from the nuclear base in a spiral around the basal body, which has two central microtubules, and axoneme for approximately 1.1 µm. The two mitochondrial derivatives began roughly at the same level and at the base of the centriole adjunct. In cross-section, they were symmetrical, with a slightly oval shape and a smaller diameter in comparison to the axoneme. The latter, also spiraled, consisted of 9 + 9 + 2 microtubules that was formed from the basal body situated just below and aligned with the nucleus. The E. polistis sperm showed the same basic structures and morphological characteristics as observed in other Chalcidoidea. However, it was possible to distinguish the sperm of this species from those of other Eulophidae by (i) the long length of the centriole adjunct on the flagellum, and (ii) the presence of two central microtubules within the basal body. The sperm characteristics suggest that Eulophidae is closely related to Trichogrammatidae and both families are more similar to Eurytomidae, Pteromalidae, and Torymidae than Agaonidae.

The peculiar structure of the flagellar axoneme in Coccinellidae (Insecta-Coleoptera).

The ultrastructure of the complex organisation of the spermatozoa in Harmonia axyridis and Adalia decempunctata (Coccinellidae) was studied, with particular emphasis on the origin of the anterior shifting of the axonemal structure, which becomes parallel to the nucleus in the sperm flagellum. In studying the spermiogenesis, a centriolar remodelling was observed with the long centriole, present in the early spermatids, transformed in the spermatozoa into an exceptionally long and narrowed basal body (about 0.16 × 3.5-4.0 µm long) displaying a 9 + 0 microtubular pattern in the proximal part and a 9 + 2 pattern in the following part; this is a characteristic not observed in any other pterygotan insect. The sperm also have a very long acrosome surrounded by a dense layer of material extending along the whole basal body. These two uncommon features were discussed in the light of sperm movement.

Re-description of the sperm morphology of Dalbulus maidis (Hemiptera: Cicadellidae: Deltocephalinae).

The corn leafhopper, Dalbulus maidis, which is one of the most important vectors of maize diseases, had the morphology of its spermatozoa described more than 40 years ago. However, the description is imprecise, requiring certain rectifications. Hence, we re-describe the sperm morphology of this species using light microscopy and transmission electron microscopy. Their spermatozoa measured from 118.1 to 128.5 µm in length, this value being about three times smaller than what was previously described. The head region is formed by a small acrosome and the nucleus. The acrosome is paracrystalline and has the bifurcated base that is docked in two cavities on one side of the anterior region of the nucleus. The nucleus, which is linear and thin, measured from 19.3 to 22.9 µm in length, and exhibited homogeneously compact chromatin. In cross-section, it has a thumbtack shape, except in the anterior and posterior regions, where it is oval. The flagellum consists of an axoneme with 9 + 9 + 2 microtubules, two similar mitochondrial derivatives, two accessory bodies each associated with a small sub-ellipsoidal structure, and a 'center-flagellar material'. In all the spermatozoa observed, the final portion of the flagellum was branched into four filaments. Some of these morphological features are now being re-described, while others have been observed for the first time in the Cicadellidae family, as the center-flagellar material.

Fine structure of the ladybird spermatozoa (Insecta, Coleoptera, Coccinellidae).

The sperm structure of several ladybird species belonging to different subfamilies of Coccinellidae was studied. Three main sperm types were clearly recognized, and were characterized by differences in acrosomal length, the presence of a dense coat around the acrosome, the length of the basal body, the amount of the centriole adjunct material, and the diameter of the mitochondrial derivatives. However, the whole group shares a pattern of the posterior sperm region uncommon for insects, in which the axoneme and other flagellar components are running parallel with the nucleus. As a general conclusion, this study has revealed an inconsistency between the sperm structure and the systematics of the group, indicating that the generic concepts within the group do not reflect a natural classification, a statement also shared by molecular studies.

Male reproductive tract and spermatozoa ultrastructure in the grasshopper Orphulella punctata (De Geer, 1773) (Insecta, Orthoptera, Caelifera).

Identification Orphulellini grasshoppers (Acrididae: Gomphocerinae) species has been difficult due to high polymorphism rate. Orphulella Giglio-Tos, 1894 is a genus with widespread geographical distribution and poor descriptions. Orphulella punctata (De Geer, 1773) has an extensive record of occurrence and available information about the phallic complex, however, there is poor data describing other parts of the male reproductive tract. The objective of this study was characterizes the internal organs of the male reproductive system and spermatozoa of O. punctata. Orphulella punctata testes are of Fountain type, each having only four follicles. Spermatozoa into the seminal vesicle are arranged in bundles with c.a. 2320 µm length, with a nucleus 110 µm long. The spermatozoa are covered by a glycocalyx, the nucleus is cylindrical with condensed chromatin and connected to the flagellum by a dense and lamellar centriole adjunct. The axoneme have 9 + 9 + 2 pattern and present two symmetrical mitochondrial derivatives. A fibrous net and two flat membranous cisternae fill the space between the axoneme and mitochondrial derivatives. This is the first description of the reproductive system of a Gomphocerinae representative.

Sperm morphology of predatory pirate bugs Amphiareus constrictus and Blaptostethus pallescens (Heteroptera: Anthocoridae) with phylogenetic inferences.

The sperm morphology of two predatory bugs Amphiareus constrictus and Blaptostethus pallescens, representing the tribes Dufouriellini and Blasptostethini, respectively, was described using light and transmission electron microscopy. The spermatozoa of Amphiareus constrictus and Blaptostethus pallescens are fine and long, each measuring 216.6µm and 181.0µm in length, of which 37.0µm and 11.6µm, respectively, comprise the nuclei. When stained with DAPI (for DNA), the posterior half of the nucleus in B. pallescens exhibited low fluorescence, while in A. constrictus this feature was observed only in the last 6µm. In both species, as in Heteroptera in general, the spermatozoa have, in the head region, an acrosome and nucleus, and in the flagellar region, an axoneme with 9 accessory tubules, 9 peripheral doublets and 2 central microtubules (9+9+2 microtubules), 2 mitochondrial derivatives (MDs), and a centriolar adjunct in the nucleus-flagellum transition. However, unlike most Heteroptera, in these species, the MDs are asymmetric, and the centriolar adjunct is quite long and encompasses completely the posterior nuclear end and the anterior tips of the MDs. These features are considered as derived, thus supporting the condition derived of Anthocoridae within Cimicomorpha. In addition, several traits of the spermatozoa of these two species easily distinguish one species (and probably a tribe) from the other; for example, the difference of formats in the MDs, and the long anterior projection of the centriolar adjunct parallel to the nucleus in B. pallescens.

Evidence of a procentriole during spermiogenesis in the coccinellid insect Adalia decempunctata (L): An ultrastructural study.

We studied spermatogenesis and spermiogenesis in Adalia decempunctata (L), a beetle of the Coccinellidae family. The spermatocyte exhibits two centrioles which elongate to form a pair of primary cilia. A novel structure, appearing in cross sections as a dense droplet, is observed near the long centriole during spermiogenesis, and is soon accompanied by a procentriole (PCL). PCL structure consists of singlet microtubules, a central tubule and an incomplete cartwheel. The PCL persists until the end of spermiogenesis, when it vanishes together with the dense droplet. The sperm has an exceptionally long basal body and the nucleus is disposed parallel to the flagellar components, a peculiar trait shared by other species of the coccinellid group. The presence of a procentriole suggested by the use of antibodies is discussed.

Testicular, spermatogenesis and sperm morphology in Martarega bentoi (Heteroptera: Notonectidae).

The testicular, spermatogenesis and sperm morphology of the backswimmer Martarega bentoi was described using light and transmission electron microscopy. In this species, a pair of testes, two deferent ducts, two different pairs of accessory glands, and an ejaculatory duct form the male reproductive system. Each testis consists of two testicular follicles, which are arranged side by side in snail shape. The follicles are filled with cysts at different stages of spermatogenesis, but in the same cyst the germ cells (up to 64) are in the same stage. At the end of spermatogenesis, the sperm cells are very long, with the flagellum measuring approximately 2500 µm in length, the nucleus only 19 µm, and the acrosome, with two distinct regions, 300 µm. The flagellum is composed of an axoneme, with a 9 + 9 + 2 microtubular pattern, and 2 asymmetric mitochondrial derivatives (MDs). These have the anterior ends inserted into two cavities at the nucleus base, exhibit two paracrystalline inclusions, and have bridges linking them to the axoneme. Few spermatozoa per cyst, asymmetry in size and shape of the MDs, as well as their insertion at the nuclear base are characteristics considered derived, and that differentiate the sperm of M. bentoi from those of the Nepomorpha, Belostomatidae and Nepidae.