Spermatophore transfer in Euchaeta species in a 2000 m water column

The numbers and placement of spermatophores transferred by males to females in the genus Euchaeta are examined in the northeastern Atlantic Ocean. Multiple placement of spermatophores is present in the five diel, vertically migrating species, E. acuta, E. pseudotonsa, E. gracilis, E. norvegica and E. hanseni. Direct placement covering the genital opening is usually, but not always the primary placement site. Males produce two sizes of spermatophores corresponding to the small directly placed and the large indirectly placed ones attached to the females. Females of the other species, E. barbata, E. barbata f. farrani, E. scotti, E. bisinuata, E. sarsi, E. abbreviata, E. longissima and E. bradyi, all meso- or bathypelagic non-vertically migrating species, have a single spermatophore attached to the genital opening. Spermatophore size is related to male and female prosome lengths and to the depth at which the species lives. Size variation of spermatophores within species decreases with increasing depth. Spermatophore transfer in deep-sea species, a single spermatophore directly placed, appears to be more efficient than in the epipelagic and mesopelagic vertical migrators where multiple placement involves a proportion of spermatophores with no direct connection to the genital opening.     

Insemination of the monogenean Neobenedenia girellae (Capsalidae,Benedeniinae)

In vitro spermatophore formation and insemination of Neobenedenia girellae (Monogenea: Capsalidae, Benedeniinae) were recorded on video and described for the first time. Upon contact of two individuals, the anterior adhesive discs of the donor firmly attached to the dorsal tegument of the recipient and the donor's fore body strongly contracted such that the genital pore region protruded and the penis was pushed anteriorly to protrude through the genital pore. It is hypothesised that the donor penis mechanically damaged the tegument of the recipient. The sperm and spermatophore matrix were released together through the penis, which was placed under the left anterior attachment disc immediately behind the adhesive pad. The spermatophore matrix containing the spermatozoa became solid and attached to the dorsal surface of recipient's body. When observed under scanning electron microscopy, the spermatophores were irregularly shaped, with a diameter of 52–83 μm. Under light microscopy they consisted of a proximal eosinophilic matrix portion and a distal thin-walled portion containing spermatozoa. Both parts were enclosed with a thin outer casing. Insemination occurred during and after spermatophore formation. Three types of insemination were recorded, unilateral and mutual insemination and self-insemination. The presence of self-insemination indicates that even a single N. girellae on a cultured fish may cause a significant parasite infection in the entire aquaculture system.     

Description of the male reproductive system of Paguristes eremita (Anomura,Diogenidae) and its placement in a phylogeny of diogenid species based on spermatozoal and spermatophore ultrastructure

The male gonopores, male reproductive apparatus, spermatophore and spermatozoa of the Mediterranean hermit crab Paguristes eremita are described, using interference phase microscopy, scanning electron microscopy and transmission electron microscopy. A correlation is made between the gonopore morphology and the different kinds of setae accompanying them, and the reproductive biology of these crabs. Each testes merges into a tubular duct made up of four zones: (1) the collecting tubule with free spermatozoa; (2) the proximal zone, where the ampulla of the spermatophores starts to be formed; (3) the medial zone, where the ampulla is completed, the stalk lengthens and the pedestal is formed; (4) the distal zone, where the mature spermatophores are stored. The sizes of the different parts of the spermatophore and of the sperm are given and their exterior morphology and ultrastructure described and compared to congeners. The morphology of the gonopore, male reproductive system, spermatophore and spermatozoa of P. eremita are species-specific, clearly distinguishing the species from the other members of the family. The available spermatozoal and spermatophore data is used to place P. eremita within a sperm phylogeny of the hermit crab family Diogenidae.     

On the occurrence of acid mucopolysaccharides in the spermatophores of two marine prawns,Penaeus indiens (Milne-Edwards) and Metapenaeus monoceros (Fabricius) (Crustacea: Macrura)

In marine prawns, spermatophores aid in sperm transfer. The prawn spermatophores, in general, consist of two parts, sperm sac and wing. The former encloses spermatozoa and the latter serves as an adhesive pad during transfer to the female's thelycum. This paper reports on the occurrence of acid mucopolysaccharides (AMPS) in the spermatophores of two penaeid prawns, Penaeus indicus (Milne-Edwards) and Metapenaeus monoceros (Fabricius). They, being the principal component of the spermatophores, have been characterized and quantified in the wing, sperm mass, sperm sac, and crystalline structures. In both prawns, the spermatophore is a simple structure. In P. indicus, it consists of sperm mass, enclosed within sperm sac, to which is attached the foliacious wing. In M. monoceros, the freshly extruded spermatophore consists of sperm mass and milky-white crystalline structures. Isolation and purification of AMPS resulted in the resolution of two toluidine blue-positive AMPS fractions in agarose-gel electrophoresis. Densitometric study of the standard and sample AMPS fractions reveals further that the two fractions correspond to chondroitin sulfate and hyaluronic acid. Quantitative assay on total AMPS shows that the sperm sac of P. indicus contains the maximum amount of AMPS (195.50 μg/mg), whereas its wing contains the least quantity of AMPS (43.68 μg/mg). The qualitative and quantitative variations found in the AMPS content of spermatophoric components of two prawns have been discussed in relation to their adhesion to thelycum as well as sperm storage and maintenance pending fertilization.     

Studies on in vitro Extrusion and Ultrastructure of the Spermatophore in Haemaphysalis longicornis (Acari: Ixodidae)

Copulation in ticks is completed by the insertion of the spermatophore into the female genital aperture by a male. The endospermatophore, a cord-like structure and contents are packed in the ectospermatophore of the completed spermatophore. The endospermatophore extrudes just after insertion of the tip of the spermatophore. Only the endospermatophore enters the female genital tract, and the ectospermatophore remains outside the female body. The extrusion is observed in vitro in Haemaphysalis longicornis at various concentrations of NaCl solution: the process is accelerated in less concentrated solutions. The cord-like structure and the endospermatophore finally receive contents extruded from the ectospermatophore. The tip of the cord-like structure connects to the surface of the endospermatophore, and together form a loop after extrusion. Ultrastructural observations confirmed that the ectospermatophore wall is composed of four layers, and the contents consist of male germ cells and three types of secretions from the male accessory genital glands. As in other ticks the male germ cells are elongated spermatids in spermatophores just after formation and extrusion. Adlerocysts described in other ticks are not found in the spermatophore of H. longicornis.     

Mode of transfer of spermatozoa in Orthoptera Tettigoniidae

A morphological and ultrastructural study was carried out on the spermatophore and spermatodoses of some species of Orthoptera Tettigoniidae. From the results concerning the spermatophore it emerged that this structure has a morphological and ultrastructural organization represented by a dilated ampulla and a peduncle or neck. From the examination of freshly deposited spermatophores and those at various time intervals thereafter, it was seen that these structures other than allowing gamete transfer, represent the site where spermatodesms, organized in the male genital tracts, undergo reorganization to acquire their definitive morphological and structural characteristics as found in the female genital tracts. The spermatodoses, in the same way as the spermatophore, represent capsules containing spermatodesms, which are originated in the spermatheca, their specific morphology seems to diversify according to the species considered. As regards their role, it is hypothesized that these structures represent a long-term conservation mechanism for spermatozoa inside the seminal receptacle.     

Spermatophore formation in the pseudoscorpion Chthonius ischnocheles (Chthoniidae)

The reproduction of pseudoscorpions involves indirect sperm transfer by means of spermatophores. The spermatophores are the product of the male genital atrium. A functional interpretation of spermatophore formation in Chthonius ischnocheles is based on evidence from (a) a morphological study of the genital atrium, the associated accessory glands and the musculature (b) males sectioned during spermatophore production (c) histochemical tests on the glands and their secretions (d) biochemical analyses of one gland (posterior dorsal) and its secretion (e) the behaviour of males during this process and (f) the structure of the spermatophore.
The anterior region of the genital atrium is concerned with the production of the sperm packet. The encysted sperm and the seminal fluid from the prostatic reservoir are encapsulated in a sperm packet by a secretion from two pairs of anterior glands. The posterior region of the genital atrium is responsible for the formation of the spermatophore stalk and its distal elaboration, the two lateral collars. These parts of the spermatophore arise from the fibroin secretion of the posterior dorsal gland; the shape of the spermatophore collars is correlated with their mould, the medial diverticula. In addition the lateral glands secrete a light oil which accumulates on a thickening of the spermatophore stalk proximal to the collars. This suggested that this secretion acts as a pheromone to attract females to the spermatophore since in this species males produce their spermatophores in the absence of females.     

Oriented spermatozoa in the spermatophore of the palaemonid shrimp, Macrobrachium rosenbergii

Estimates of the degree of orientation of spermatozoa within spermatophores of the freshwater shrimp, Macrobrachium rosenbergii, were made using electron micrographs of ultrathin sections cut transverse and parallel to the longitudinal axis of the spermatophores. Counts of the total number of longitudinal and non-longitudinal profiles of sperm spikes in transverse sections of spermatophores indicated that 67% of the spermatozoa were oriented with their spikes approximately perpendicular to the long axis of the spermatophore. In longitudinal sections, ~30% of the sperm spike profiles were oriented parallel to the longitudinal axis of the spermatophore, leaving ~70% of the profiles oriented in planes not parallel to the longitudinal spermatophore axis. Stereologic analyses based on the number of intersections of longitudinally sectioned sperm spike profiles per unit length of test lines placed over photographic images of ultrathin sections cut in both transverse and longitudinal planes of spermatophores gave a Saltykov approximation of 0.607, favoring the view that the spermatozoa were oriented with their spikes perpendicular to the long axis of the spermatophore. This orientation brings many of the sperm bases approximately parallel to the surface of the spermatophore, an orientation which may facilitate normal sperm-egg interaction during fertilization in this species.     

The reproductive biology of Euchaeta antarctica Giesbrecht (Copepoda: Calanoida) at South Georgia

The reproductive biology of the predatory calanoid copepod Euchaeta antarctica Giesbrecht was investigated in two interconnected fjord systems at South Georgia. Counts of the number of spermatophores attached to adult females and the number of egg sacs encountered, indicated probable peaks of reproduction in summer and winter. Patterns of spermatophore placement were examined and compared with data for E. norvegica (Boeck) from boreal waters. Elemental analysis indicated a high proportion of carbon and a low proportion of nitrogen in adult females and egg sacs from both sites.High winter carbon levels in adults seem related to their predatory feeding habits allowing high food intake throughout the year whereas in egg sacs it probably reflects extended development times and/or non-feeding naupliar stages.Mean adult female and egg clutch dry weights were higher in Cumberland East Bay during winter than in Moraine Fjord during summer. These differences are discussed in the context of relationships between fjord morphology and production levels.     

Description of the male reproductive system of Paguristes eremita (Anomura, Diogenidae) and its placement in a phylogeny of diogenid species based on spermatozoal and spermatophore ultrastructure

The male gonopores, male reproductive apparatus, spermatophore and spermatozoa of the Mediterranean hermit crab Paguristes eremita are described, using interference phase microscopy, scanning electron microscopy and transmission electron microscopy. A correlation is made between the gonopore morphology and the different kinds of setae accompanying them, and the reproductive biology of these crabs. Each testes merges into a tubular duct made up of four zones: (1) the collecting tubule with free spermatozoa; (2) the proximal zone, where the ampulla of the spermatophores starts to be formed; (3) the medial zone, where the ampulla is completed, the stalk lengthens and the pedestal is formed; (4) the distal zone, where the mature spermatophores are stored. The sizes of the different parts of the spermatophore and of the sperm are given and their exterior morphology and ultrastructure described and compared to congeners. The morphology of the gonopore, male reproductive system, spermatophore and spermatozoa of P. eremita are species-specific, clearly distinguishing the species from the other members of the family. The available spermatozoal and spermatophore data is used to place P. eremita within a sperm phylogeny of the hermit crab family Diogenidae.     

When do the Costs of Spermatogenesis Constrain Sperm Expenditure? Remarks on the Pattern of the Spermatogenic Cycle

The costs of spermatogenesis constrain sperm expenditure when sperm production per day is limited. Thus, males are challenged to allocate available resources to sperm production and other life history functions. However, this prevailing assumption is not applicable to species in which spermatogenesis becomes quiescent during the breeding season. Males of these species prepare large quantities of sperm before the breeding season. Among these species, constraints on ejaculates have been intensively investigated in salamanders that deposit spermatophores. Although it is predicted that sperm expenditure should not be limited because of abundantly prepared sperm, spermatophore deposition is often limited during the breeding season when vas deferens are full of sperm. We tested a hypothesis regarding limited spermatophore deposition by measuring sperm quantity and volume of spermatophores sequentially deposited by male eastern newts Notophthalmus viridescens. A male newt rarely deposits more than three spermatophores per mating. If depletion of non‐sperm components of spermatophores limits spermatophore deposition, we predicted that spermatophore volume decreases while sperm quantity remains constant as a male deposits more spermatophores. Alternatively, some regulative mechanisms allow a limited portion of available sperm to be expended per mating, in which sperm quantity is predicted to decrease while the spermatophore volume remains constant. Finally, depletion of non‐sperm components may regulate sperm expenditure, which predicted that both spermatophore volume and sperm quantity decrease. We found that both sperm quantity and the spermatophore volume decreased as a male deposited more spermatophores during a single mating. Sperm expenditure was constrained without the costs involved in active spermatogenesis, and depletion of non‐sperm components likely regulate sperm quantity loaded in spermatophores. In dissociated spermatogenesis, constrained sperm expenditure do not mean that costly spermatogenesis is directly limiting male mating capacity but rather suggest that the evolution of physiological mechanisms regulating sperm expenditure per mating maximizes male reproductive success.     

Spermatangium formation and sperm discharge in the Japanese pygmy squid Idiosepius paradoxus

In cephalopods, sperm discharge is an important event not only for sperm transfer but also influencing sperm storage capacity of attached spermatangia (everted spermatophores). To investigate sperm discharge from spermatangia and the condition of naturally attached spermatangia in Japanese pygmy squid (Idiosepius paradoxus) we (i) investigated the morphology of spermatophores and spermatangia, and the process of spermatophore evagination and sperm discharge from spermatangia obtained in vitro; (ii) observed spermatangia that were naturally attached to female squids at 6, 12, 18, 24 and 48 h after copulation to investigate alterations in naturally attached spermatangia with time. The spermatophore of I. paradoxus is slender and cylindrical and consists of a sperm mass, a cement body and an ejaculatory apparatus, which is similar to those of loliginid squids. The spermatangium is fishhook-shaped, its distal end being open and narrow. After the spermatangium is formed, the sperm mass gradually moves to the open end of the spermatangium, from where sperm are released. Sperm discharge is a rapid process immediately after the beginning of sperm release, but within 5 min changes to an intermittent release of sperm. Although the volume of residual spermatozoa differed among spermatangia that were naturally attached to a single individual, the probability that spermatangia would be empty increased with time. Most naturally attached spermatangia discharged almost all of their spermatozoa within 24 h after copulation, and no spermatangia were attached to females 48 h after copulation. These results suggest that sperm transfer from the spermatangium to the seminal receptacle must occur within 24 h, and that the spermatangium functions as a transient sperm storage organ in pygmy squids.     

Repeated visitations of spermatophores and polyandry in females of eriophyoid mites

Eriophyoid females store sperm either asymmetrically in one spermatheca, or symmetrically in both spermathecae. Previous studies have suggested that species in which females store sperm asymmetrically pick up sperm from only one spermatophore, while those with symmetrical sperm storage pick up sperm from two or more spermatophores during their lifetime. The aim of this study was to examine spermatophore visitation behaviour and symmetry of sperm storage in Aculops allotrichus from the black locust tree and Cecidophyopsis hendersoni from the yucca. This would indicate monandry or polyandry in these species. In both eriophyoids, the spermatophore visitation consisted of three phases: mounting, lying on the spermatophore and dismounting. Aculops allotrichus stored sperm asymmetrically. However, nearly one-third of the observed females visited two spermatophores, rather than only one in their lives. When A. allotrichus females visited two spermatophores they spent a similar amount of time at the first and at the second visitation. Also, the times of visitation of the first of the two spermatophores and the single spermatophore in a female lifetime did not differ significantly. This would suggest that apart from monandry, double insemination also occurs in this species. By contrast, C. hendersoni females were polyandrous. They stored sperm symmetrically and visited several spermatophores, on average 1.54 (max 6) per day, and up to 33 spermatophores in their lives. The benefits of repeated spermatophore visitation and the possible mechanisms of sperm storage in both species are discussed.     

Morphological and morphometric appraisal of the spermatophore of the southern hermit crab Isocheles sawayai (Anomura: Diogenidae), with comments on gonopores in both sexes

The spermatophore morphology of the hermit crab Isocheles sawayai from southwestern Atlantic (Brazil) is described. The spermatophores show similarities with those described for other members of the family Diogenidae, especially with the recently described Loxopagurus loxochelis. The spermatophore is composed of three major regions: a sperm filled head or ampulla, a columnar stalk and a foot or pedestal. The spermatophores show specific morphology in having a circular ampulla, and a constriction or neck between the ampulla (100 μm) and the thin (27 μm), long stalk (500 μm). The stalk penetrates less than half way into the spermatophore head. Most spermatophores show one of the small posterior projections on the underside of the ampulla as being bigger than the other, making it asymmetrical. The size of the spermatophore is related to hermit crab size with direct relationships found between spermatophore ampulla width, total length, and peduncle length with shield length of the hermit crab. The morphological characteristics of the spermatophore of I. sawayai are species-specific distinguishing it from other members of the family, and are useful to infer further phylogenetic relationships.     

Spermatophores of the salamander Ambystoma texanum

Living spermatozoa were observed in freshly deposited spermatophores and in fluid from vasa deferentia. In the distal, but not proximal, vas deferens spermatozoa moved together in whorls with heads and tails in alignment. Around the entire periphery of the spermatophore cap, similar slowly undulating groups of spermatozoa had their heads aligned and directed outward. Over time, some individual spermatozoa left the cap of the spermatophore and moved into the surrounding water (cap deterioration). Microscopical observations were made on spermatophore squashes and paraffin sections of spermatophores and vasa deferentia. Spermatozoa around the periphery of the cap were underlain by a PAS-positive membrane-like material. Cytoplasmic droplets, which were attached to spermatozoan necks in the vas deferens, were accumulated in the center of the spermatophore cap deep to the PAS-positive membrane. Spermatophore stalks were strongly PAS and Alcian blue positive and showed positive reaction for RNA. Tests for lipids and proteins were negative in the whole spermatophore. Electron microscopic observations showed the stalk of the spermatophore to be composed of rounded ‘balls’ of fibrous material. At the juncture of the stalk and cap a less dense fibrous material impacted the stalk enclosing many sperm tails and some heads and, although no attachment devices were visualized, the sperm were closely apposed to this material as was the spermatophore stalk. This finely filamentous material encircled the cap and was more prominent in some regions than others. The PAS-positive material detected with the light microscope was also observed with the electron microscope. It was circumferentially oriented and was composed of 200 Å packed filamentous densities. Sperm heads and tails were found lying external to the membrane, whereas only tails and cytoplasmic droplets occupied the core of the spermatophore. Cytoplasmic droplets were usually free of the sperm tail and contained membranous sacs and two types of nuage material.     

Daily production of spermatophores, sperm number and spermatophore size in two eriophyoid mite species

Under dissociated sperm transfer, (non-pairing) males deposit spermatophores on a substrate, while females seek spermatophores and pick up sperm on their own. Spermatophore expenditures of non-pairing males should be high, due to the increased uncertainty of sperm uptake by a female. In this study I examined spermatophore expenditures in two eriophyoid species that differed in the degree of dissociation between sexes: (1) Aculus fockeui (Nalepa and Trouessart) males rarely visit quiescent female nymphs (QFNs), and mostly deposit spermatophores all over the leaves, whereas (2) Aculops allotrichus (Nalepa) males guard QFNs for many hours and deposit several spermatophores beside them. Males of both species were collected from the field and tested in solitude. Aculus fockeui males deposited on average 19.1 spermatophores per day, whereas A. allotrichus deposited only 3.6 spermatophores per day, and had a very large coefficient of variation. Males and spermatophores of A. allotrichus were significantly smaller and contained less sperm than those of A. fockeui. In both eriophyoids, spermatophore size was fitted to the size of female genitalia and the height of females. The ratio between the diameter of spermatophore head and the width of a female genital coverflap was 0.6, whereas the ratio between the female leg and the length of spermatophore stalk was 0.5. Several factors could be responsible for the discrepancy in spermatophore expenditures between species. Among other factors, the effects of male size, male reproductive strategy and female genitalia size on spermatophore output and size of spermatophores are discussed.     

Spermatophore transfer in the hermit crab Clibanarius vittatus (Crustacea,Anomura, Diogenidae)

Although mating has been described in several hermit crab species, the mechanics of spermatophore transfer have not previously been demonstrated. Evidence from pleopod and gonopore morphology, video observations, and inseminated females indicates that in Clibanarius vittatus the male applies a spermatophoric mass directly onto the female via the gonopores rather than with modified pleopods 1-2 (gonopods) and/or genital papillae as in many other decapods. The single second pleopod of males of C. vittatus has a simple endopod with no apparent modifications for sperm transfer. There are no genital papillae extending from the male gonopores. The globular spermatophores are aligned in rows surrounded by a seminal secretion in the male ducts (vasa deferentia that terminate in ejaculatory ducts opening to the exterior via the gonopores). During copulation, described from time-lapse video recordings, the ventral surface of the last thoracic segment of the male, bearing the gonopores, was apposed to the ventral cephalothorax of the female. A massive amount of seminal secretion containing spermatophore ribbons, termed here the spermatophoric mass and described for the first time in a hermit crab species, was observed covering the sternites and coxae of pereopods 1-5 of a recently copulated female. It is suggested that during copulation the male emits the contents of the ejaculatory ducts directly onto the female without the aid of gonopods or genital papillae. Although spermatophore transfer is simple in C. vittatus, the presence of modified anterior pleopods or elongate genital papillae (sexual tubes) in other paguroidean species suggests the possibility of a more complex insemination process in these other hermit crabs.     

Spermatophore formation and sperm ultrastructure of Sundathelphusa philippina (Crustacea: Brachyura: Gecarcinucidae)

We investigated the morphology of spermatozoa, spermatophores and the anterior vas deferens (AVD) of the gecarcinucid freshwater crab Sundathelphusa philippina. The morphology of the acrosome (proportions, structure and arrangement of acrosomal layers) and the spermatophores complies with the known sperm and spermatophore morphology of the brachyuran family Gecarcinucidae. The sperm cells are packed within coenospermic spermatophores that are of a mucous type, lacking a complex spermatophore wall. Spermatophore formation takes place in the distal part of the AVD. The strongly proliferated inner epithelium of the vas deferens releases vesicles via apocrine secretion. These vesicles fuse with the incipient spermatophores that subsequently coalesce, thus forming the coenospermic aggregates that represent the mature spermatophores.     

Experimental evolution reveals divergence in female genital teeth morphology in response to sexual conflict intensity in a moth

The rapid evolutionary divergence of male genital structures under sexual selection is well documented. However, variation in female genital traits and the potential for sexual conflict to drive the coevolution between male and female traits has only recently received attention. In many lepidopterans, females possess genital teeth (collectively, signa). Comparative studies suggest these teeth, involved in the deflation of spermatophores, may have coevolved with male spermatophore thickness via sexually antagonistic coevolution in a contest over the rate of deflation of spermatophores within the reproductive tract. We tested the hypothesis that sexual conflict should generate coevolution between genital teeth and spermatophore morphology by examining these traits under experimental manipulation of sexual conflict intensity. Using micro‐CT scanning, we examined spermatophore and teeth morphology in populations of the Indian moth, Plodia interpunctella, which had been evolving for 110 generations under different adult sex‐ratio biases. We found divergence in female signa morphology in response to sexual conflict: females from female‐biased populations (reduced sexual conflict) developed wider signa. However, we found no evidence of coevolution between signa traits and spermatophore thickness as reported from comparative studies.     

The effect of the presence of quiescent female nymphs, males and their spermatophores on spermatophore placement in two species of eriophyoid mites

Under sex dissociated sperm transfer, females seek spermatophores and pick up sperm without male assistance. In several species males adjust spermatophore deposition rate to the presence of conspecifics. It is not known, however, which factors could favor such elasticity in non-pairing males. In this paper, we compare male response towards conspecifics between the sex dissociated eriophyoid mites Aculus fockeui (Nalepa and Trouessart) and Aculops allotrichus (Nalepa). The species differ significantly in male reproductive strategies and, consequently, the intensity of male–male-competition. Aculus fockeui males deposit spematophores all over the leaves and occasionally leave single spermatophores beside quiescent female nymphs (QFNs). In contrast, A. allotrichus males guard QFNs and encircle them with spermatophores. In this study, males of both species deposited spermatophores close to and apart from the rival spermatophores. Aculops allotrichus males had similar spermatophore output whether they were kept alone or in a group of seven males. They did not change spermatophore output in the presence of five rival spermatophores, a QFN or a QFN and varying number of rivals, either. In contrast, A. fockeui males increased spermatophore output in the presence of rival spermatophores or when on the arena with a QFN the male number increased to eight males. They did not respond, however, to the presence of a QFN and one rival or a QFN alone. The possible effect of the species-specific intensity of male–male competition, population density, the availability of receptive females and the rate of spermatophore output on the flexibility of eriophyoid spermatophore deposition is discussed.