Sperm drinking by female catfishes: a novel mode of insemination

Synopsis We report on unique reproductive behaviour and a new mode of egg insemination in a small catfish Corydoras aeneus (Callichthyidae). A male courts a female by presenting his abdomen to her. Before releasing eggs, the female attaches her mouth to the male's genital opening and directly drinks his sperm. The sperm pass through her intestine and are discharged together with eggs into the pouch formed by her pelvic fins. Thus, eggs are mixed with fresh non-dispersed sperm in an enclosed space, ensuring effective insemination. This mode of insemination is novel to fishes, but is likely not restricted to catfishes of the genus Corydoras.     

Ultrastructure of the spermatozoa and eggs of the ocean pout (Macrozoarces americanus L.), an internally fertilizing marine fish

Ultrastructure of sperm and eggs of the ocean pout (Macrozoarces americanus L.), an internally fertilizing marine teleost, was examined by scanning and transmission electron microscopy. The results showed that the sperm do not have an acrosome but have a very long mid-piece (one to two times the sperm head length) containing numerous well-developed elongated mitochondria. The sperm also have two tails (is biflagellate) each consisting of nine peripheral and one central pair (9 ± 2) of microtubules. This long mid-piece and the biflagellate nature of the sperm appear to be associated with the long life-span of the sperm and with sperm dispersal in the ovary to fertilize the eggs internally. The ocean pout eggs are enveloped by a porous chorionic membrane similar to that found in other teleosts but have two micropyles, a condition likely related to a mechanism of egg fertilization which increases the egg fertlity in the presence of low sperm numbers. Following insemination, some biochemically undefined excretions appeared on the surface of fertilized eggs and led to the acquisition of adherent capability of the eggs which formed a tightly associated egg mass in sea water. © 1995 wiley-Liss, Inc.     

Comparison of heterospermic and homospermic inseminations as measures of male fertility

This study attempted to determine a basis for the previously observed greater sensitivity of heterospermic tests when compared to homospermic tests for detecting differences in fertility between males. In theory, the results of heterospermic tests are an indication of the proportion of eggs fertilized per unit time whereas results of homospermic inseminations measure only the cumulative or final proportion of eggs fertilized. The fertilizing ability of sperm from males of CF1 and C57BL/6N strains of mice was compared homospermically using both relatively high and low concentrations of sperm and by measuring the proportion of eggs penetrated per unit of time. The fertilizing ability of sperm from these strains was also compared using heterospermic inseminations. When females were inseminated with a high concentration of sperm, males of both strains fertilized a high and indistinguishable percentage of eggs when examined after 30 hr. When females were inseminated with either a low concentration of sperm or when the proportion of eggs penetrated was measured at 5 hr, differences between strains of mice were distinguishable. Heterospermic insemination further magnified the observed difference between strains. The results of this study confirm that measuring the percentage of eggs fertilized per unit of time can enhance the magnitude of differences between males in fertility as compared to measuring only the final percentage of eggs fertilized. Measuring the percentage of eggs fertilized per unit of time does not, however, entirely account for the large differences observed between fertility of males when they are compared using heterospermic inseminations.     

RELATIONSHIPS BETWEEN SPERM LENGTH AND SPEED DIFFER AMONG THREE INTERNALLY AND THREE EXTERNALLY FERTILIZING SPECIES

It is often assumed that longer sperm, by virtue of their increased swimming speed, have a fertilization advantage over shorter sperm when in competition to fertilize eggs. However, there is surprisingly little evidence for a positive correlation between sperm length and speed. Here we use an approach that accounts for within‐male variation in sperm traits to examine the relationships between sperm length and sperm speed across a broad range of species, including three internally fertilizing species and three externally fertilizing species. Our results reveal that correlations between sperm size and speed are indeed present and possibly more common than currently thought. However, the direction of the correlations between sperm length and speed, which are more prevalent within a male's ejaculate than among males, were influenced by fertilization mode in contrasting and unexpected ways. Broadly, the patterns revealed that in externally fertilizing species sperm with longer flagellum and shorter heads relative to their flagellum swam faster, whereas in internally fertilizing species sperm with shorter flagellum and longer heads relative to their flagellum swam faster. We discuss these results in light of sperm competition theory and contrast the intraspecific patterns observed in this study with macroevolutionary patterns of sperm evolution reported elsewhere.     

Reproductive histology of Tomeurus gracilis Eigenmann, 1909 (Teleostei: Atherinomorpha: Poeciliidae) with comments on evolution of viviparity in atherinomorph fishes

Tomeurus gracilis is a species long considered pivotal in understanding the evolution of livebearing in atherinomorph fishes. Tomeurus gracilis is a zygoparous or embryoparous poeciliid: internal fertilization is followed by females laying fertilized eggs singly or retaining fertilized eggs until or near hatching. Tomeurus was hypothesized as the sister group of the viviparous poeciliids until it was proposed as a close relative of a derived viviparous poeciliid, Cnesterodon, hence nested among viviparous taxa rather than near the root of the tree. Here, we describe and compare reproductive morphological characters of the little‐known Tomeurus with those of representative atherinomorphs. In Tomeurus and Cnesterodon, sperm are packaged in naked sperm bundles, or spermatozeugmata, in a configuration considered here diagnostic of viviparous poeciliids. Testes are single and free sperm are stored in the ovary in both taxa in contrast to oviparous atherinomorphs in which testes are paired and sperm are not packaged and not stored in the ovary. Efferent ducts in Cnesterodon testes and other viviparous poeciliids have a PAS‐positive secretion demonstrating presence of a glycoprotein that inactivates sperm or prevents final sperm maturation. No PAS‐positive staining secretion was observed in Tomeurus or oviparous atherinomorphs. Tomeurus shares apomorphic reproductive characters, such as sperm bundle and testis morphology and a gonopodium, with viviparous poeciliids and plesiomorphic characters, such as a thick zona pellucida with filaments, with oviparous taxa. We do not postulate loss or reversal of viviparity in Tomeurus, and we corroborate its phylogenetic position as sister to the viviparous poeciliids. J. Morphol., 2010. © 2010 Wiley‐Liss, Inc.     

The evolution of the type I interferon gene family in mammals

A phylogenetic analysis of mammalian type I interferon (IFN) genes showed: (1) that the three main subfamilies of these genes in mammals (IFN-, IFN-, and IFN-) diverged after the divergence of birds and mammals but before radiation of the eutherian orders and (2) that IFN- diverged first. Although apparent cases of interlocus recombination among mouse IFN- genes were identified, the hypothesis that coding regions of IFN- genes have been homogenized within species by interlocus recombination was not supported. Flanking regions as well as coding regions of IFN- were more similar within human and mouse than between these species; and reconstruction of the pattern of nucleotide substitution in IFN- coding regions of four mammalian species by the maximum parsimony method suggested that parallel substitutions have occurred far more frequently between species than within species. Therefore, it seems likely that IFN- genes have duplicated independently within different eutherian orders. In general, type I IFN genes are subject to purifying selection, which in the case of IFN- and IFN- is strongest in the putative receptor-binding domains. However, analysis of the pattern of nucleotide substitution among IFN- genes suggested that positive Darwinian selection may have acted in some cases to diversify members of this subfamily at the amino acid level.     

Testing for non-randomness in sizes and habitats of West Indian lizards: choice of species pool affects conclusions from null models

Summary Several null models are proposed for testing whether size or habitat differences in West IndianAnolis lizards are greater than expected by chance. The models differ primarily in choice of the pool from which species are sampled to form random communities. Regardless of choice of pool, size differences in the Lesser Antilles are greater than null models predict; the pool using species on the known source (Puerto Rico) gives a greater variance in ratios but about the same mean ratio (for males), or a greater mean ratio (for females), compared with the pool composed of species on the islands being tested (the Stronget al., 1979, Galápagos procedure). On satellite islands of the Greater Antilles, sizes do not differ more than expected from null models. Pools composed of mainland-source species give null communities with more small or more large ratios than those composed of island species, depending upon whether four-species islands are included or excluded, respectively. Colwell and Winkler's unmodified Narcissus hypothesis is contradicted by these results in procedures where species not likely to be able to occur on small islands are included in the species pool. Using the most biologically reasonable, but not other, choices of source pool, species on satellite islands of the Greater Antilles differ more in structural habitat then expected by chance. In contrast to some of the results on size, here mainland-source pools are more likely to produce a statistically significant difference between real and random communities, as predicted by the Narcissus hypothesis. However, exclusion of structural habitat categories not found on satellite islands is necessary to achieve this significance.     

Invertebrate predator-prey body size relationships: an explanation for upper triangular food webs and patterns in food web structure?

Summary It has been suggested by Cohen and Newman (1985) that many of the patterns in published food webs can be derived from a stochastic model in which the species are arranged in a trophic hierarchy (the cascade model). We suggest that, if predators are larger than their prey, a trophic hierarchy can be generated on the basis of body size Empirical evidence from the literature shows that there is a positive relationship between predator and prey size for a range of invertebrates and that predators are usually larger than their prey. Using experimental data on an aquatic food web we show that body size can lead to the type of trophic hierarchy used in the cascade model, suggesting that many food web patterns may be a product of body size. This conclusion is discussed with respect to the limitations of the food web data and the relationship between static and dynamic models of web structure.     

Difference of fertilizing capacity between testicular sperm and vas deferens sperm in Cynops pyrrhogaster

The fertilizing capacity was compared between testicular and vas deferens sperm in Cynops pyrrhogaster. The testicular sperm was not capable of fertilizing jelly eggs. In contrast, the vas deferens sperm was already capable of fertilizing the newt jelly eggs. There was no inhibitory factor for fertilizing jelly eggs in the testis. These results suggest that the testicular sperm is immature as to the fertilizing capacity. The testicular sperm gained the fertilizing capacity for the jelly eggs by treatment with Holtfreter's solution or 1/20 strength Holtfreter's solution. The treatment may promote the step of maturation to achieve the fertilizing capacity. The treated testicular sperm did not fertilize dejellied eggs, although vas deferens sperm fertilized dejellied eggs. Therefore, the maturation state of the treated testicular sperm is different from that of vas deferens sperm. Newt sperm may be matured within the vas deferens, as the newt does not have an organ like the mammalian epididymis.     

The effects of testosterone, 5α-dihydrotestosterone, 3α-androstanediol,and 3β-androstanediol on the maturation of rabbit epididymal spermatozoa in organ culture

Summary The fertilizing ability of spermatozoa from epididymal tubules maintained in organ cultures from 1 to 7 days was assessed after artificial insemination into receptive does. It was found that spermatozoa from the distal corpus which were already capable of fertilizing eggs prior to the cultures retain this ability for 1 day without addition of hormone and for 3–4 days when testosterone (0.5 g/ml) or 5-dihydrotestosterone (0.5 g/ml) is added to the culture medium. Spermatozoa from the proximal corpus which were not capable of fertilizing eggs prior to the cultures remain so after 1 day in cultures without addition of hormone. Testosterone, 5-dihydrotestosterone, 3-androstanediol, or 3-androstanediol was added to cultures of proximal corpus at a concentration of 0.5 g/ml. Only with 5-DHT is the mean percentage of fertilization significantly higher than the percentage obtained without addition of hormone. Insulin does not potentiate the effect of 5-DHT on sperm fertilizing ability. Epithelial growth factor is ineffective. Spermatozoa from the caput epididymidis kept in cultures for 1 to 4 days remain infertile. The results are discussed in light of the morphological findings presented in the preceding communication and in relation to the physiological requirement for sperm maturation in the epididymis.     

Migration and division of cleavage nuclei in the gall midge,Wachtliella persicariae

Summary In the eggs ofWachtliella persicariae the cleavage nuclei move relative to the surrounding ooplasm. This active migration is caused by an organelle whose ultrastructure was studied throughout the mitotic cycle. It consists of a greatly enlarged polar cytaster derived from the mitotic apparatus, linked to the nucleus by 100 Å filaments. The microtubules of the cytaster were found only during periods of active nuclear migration, i.e., from the onset of anaphase to the early prophase of the next mitotic cycle. They are always solitary and follow the course of the astral rays, which are known to temporarily adhere to peripheral structures of the egg cell and to exert tractive forces. In contrast to the cytaster microtubules, the microtubules in the spindle are bundled and persist from early metaphase through late telophase.During ontogenesis the first migration cytaster is built up between 3 and 12 min after oviposition near the anterior egg pole, in the vicinity of the sperm nucleus. In non-inseminated eggs time lapse films show a migration cytaster to develop autonomously in a region free from nuclei, but it does not follow the normal path of the male pronucleus. In several cases the female pronucleus, which remains without a cytaster of its own, was observed to move to the cytaster generated in the absence of the male pronucleus. Whether or not it is adhering to a nucleus, the cytaster divides into two at the correct time, i.e, corresponding to the first cleavage division in fertilized eggs. In some non-inseminated eggs this type of pseudocleavage has been observed to occur repeatedly, giving rise to an increasing number of anucleate cytasters.     

Mycolic acid patterns of some species of Mycobacterium

Representative strains of some species of Mycobacterium were degraded by both acid and alkaline methanolysis. Two-dimensional thin-layer chromatography was used to determine the patterns of mycolic acids and other long-chain components in these methanolysates. Patterns composed of -, methoxy- and ketomycolates were found in Mycobacterium asiaticum, Mycobacterium bovix, Mycobacterium gastri, Mycobacterium gordonae, Mycobacterium kansasii, Mycobacterium marinum and Mycobacterium tuberculosis; a representative of Mycobacterium thermoresistibile also contained lower molecular weight -mycolates in addition to these three acids. In representatives or Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium nonchromogenicum, Mycobacterium novum, Mycobacterium paratuberculosis, Mycobacterium scrofulaceum, Mycobacterium terrae, Mycobacterium xenopi, and Mycobacterium sp. MNC 165 - and ketomycolates were accompanied by -carboxymycolates and 2-eicosanol and homologous alcohols which are derived from wax-ester mycolates. Mycobacterium fortuitum and Mycobacterium giae contained - and epoxymycolates and both serovars of Mycobacterium simiae had a very characteristic pattern of -, - and ketomycolic acids. Comparison with data for other mycobacteria showed the chemotaxonomic significance of these mycolic acid patterns.Abbreviations TLC thin-layer chromatography - TBDMS t-butyldimethylsilyl Supplementary data: Copies of the chromatographic patterns of the mycolic acids from all the strains examined can be provided on request from one of the authors (D.E.M.)     

Insemination patterns in HawaiianDrosophila species (Diptera: Drosophilidae) correlate with ovarian development

In 35 Hawaiian Drosophilaspecies collected from natural populations, all reproductively mature females were inseminated. Onset of receptivity to insemination broadly correlated with the vitellogenic stage of ovarian development, but species and species groups varied in the exact stage of vitellogenesis at which earliest insemination took place. A striking exception was observed in two picture-winged species of the adiastolasubgroup, which were precociously inseminated in previtellogenesis, close to the time of adult eclosion. For a given species, the range of ovarian stages from earliest receptivity to insemination of all females, termed the insemination period, also varied among species. This variability may be due to physiological variation between females in hormonal levels, low population densities, and lek behavior of males, or it may reflect variation in degree of female discrimination toward male courtship attempts. Consideration of the quantities of sperm observed stored in the female sperm storage organs, relative to the numbers of eggs produced, indicates obligatory multiple insemination for some species, notably the fungus breeders. In other species groups, there is no apparent necessity for remating, although it may occur.This paper is No. IV in the series Studies of Oogenesis in Natural Populations of Drosophilidae.     

Sperm velocity and longevity trade off each other and influence fertilization in the sea urchin Lytechinus variegatus

The theoretical prediction that fast sperm should be more effective at fertilizing eggs has never been documented empirically. Interspecific comparisons suggest an inverse relationship between sperm velocity and sperm longevity but this trade-off has never been demonstrated within a species. Here I investigate how sperm velocity and sperm longevity influence the patterns of fertilization in the sea urchin Lytechinus variegatus. In the laboratory I examined 11 male female pairs of sea urchins for variation in sperm velocity and sperm longevity, and determined the correlations of these traits with the percentage of eggs fertilized with serially diluted sperm. Males with faster sperm had higher rates of fertilization than males with slower sperm. Within individual males, as sperm aged they slowed down and showed a reduced percentage activity and lower rates of fertilization. Across males, the average velocity of freshly spawned sperm was inversely related to sperm longevity. These results establish the possibility that sperm traits are adapted for varying conditions along a continuum from sperm limitation to sperm competition.     

An explication of ‘wind illness’ in Northern Thailand

Wind illness is a very common complaint among the Northern Thai, yet is rarely recognized by Thai physicians trained in biomedicine. Persons most susceptible to wind illness are adult women who have ever borne a child. Consequently, data were obtained from 415 everparous women, 43% of whom reported ever having had wind illness and 57%, never having had it. In addition, 20 individuals who had ever had the syndrome were followed for case study, and 13 indigenous healers who traditionally treat clients suffering wind illness were interviewed. Their perceptions of the etiology, symptomatology and treatment of wind illness are reported in Part I. Part II is an attempt to define wind illness in terms of biomedicine and as a consequence of fertility. Part III synthesizes the emic and etic accounts with explanations for the perdurance of wind illness despite the advances of biomedicine and the recent fertility decline in Northern Thailand.     

The effects of testosterone, 5α-dihydrotestosterone, 3α-androstanediol,and 3β-androstanediol on epithelial fine structure of the rabbit epididymis in organ culture

Summary The fine structure of the corpus epididymidis of the rabbit has been studied following organ culture. Various modifications of tissue preparation and culture conditions were examined to obtain good maintenance of cellular integrity as well as to preserve sperm fertilizing ability.After 5 to 7 days in culture in the absence of hormonal support, the epididymal epithelium showed signs indicative of cellular regression. Such changes included shrinkage of the cells, loss of the border of stereocilia, decrease in smooth endoplasmic reticulum, and an increase in autophagic vacuoles. The presence of androgens in culture media prevented cellular regression to varying degrees, depending on the hormone utilized. With regard to maintenance of cellular integrity, potency of the androgens tested was as follows: 5-dihydrotestosterone >= 3-androstanediol > testosterone > 3-androstanediol. Addition of insulin to dihydrotestosterone-containing cultures resulted in no improvement in maintenance.Phagocytosis of spermatozoa by epithelial cells was observed in cultured tubules and the degree of spermiophagy was inversely proportional to successful maintenance of fine structural characteristics of epithelial cells.The morphological findings reported here correlate well with the fertilizing ability of spermatozoa from cultured epididymis as reported in an accompanying communication.     

Infrared analysis of eleven carrageenophytes from Baja California,Mexico

Infrared analyses of the carrageenan in ten species (representing four genera) of Gigartinaceae and one species of Hypneaceae in different reproductive phases from the northwestern coast of Baja California were studied. Cystocarpic samples of the Gigartinaceae presented varying degrees of a / hybrid. The degree of hybridization was determined based on the ratio between the peak absorbances at 805/845 cm^–1. A high correlation was observed between the 805/845 cm^–1 and 805/970 cm^–1 ratios. Tetrasporic samples of Gigartina leptorhynchos, Iridaea splendens, Rhodoglossum affine and R. roseum, presented a -carrageenan profile, whereas Gigartina tepida, G. exasperata, G. harveyana, G. canaliculata and G. spinosa presented a -carrageenan. The tetrasporic sample of Hypnea valentiae showed a -carrageenan with a very low degree of hybridization.     

Conditions for initiation of fertilization of eggs in the copulating elkhorn sculpin

The reason for failure to initiate fertilization internally was examined in a cottid fish, the elkhorn sculpin, Alcichthys alcicornis which has internal gametic association and external fertilization. While eggs could be activated in calcium free hypertonic media but not be fertilized, fertilization occurred in isotouic media rich in calcium ions. The rate of fertilization was dependent on calcium concentration, and eggs were not fertilized in solutions with a calcium ion concentration of less than 0·57 mmol kg⁻¹. Calcium ions could be replaced to some extent by magnesium ions, but the former were the more effective in fertilization. Since calcium ion concentration of ovarian fluid of A. alcicornis was 0·41 mmol kg⁻¹, it was inferred that low calcium concentration in the ovarian fluid was the cause of the failure of A. alcicornis eggs to fertilize internally.     

Life-history consequences of resource allocation of two bdelloid rotifer species

Two bdelloid species,Macrotrachela quadricornifera (aquatic species) andPhilodina vorax (terrestrial moss species), with similar survival but different age-specific fecundity schedules, were analyzed daily to determine growth rates and the volume invested in reproduction. The two species had similar growth patterns and started reproduction while still growing. In both, the size at maturity was independent of age.M. quadricornifera resumed growth after reaching a size plateau when reproduction was over, whileP. vorax continued to reproduce until death. Although the net reproductive rate ofP. vorax was consistently lower than that ofM. quadricornifera, the same percent of adult volume was invested in reproduction over its life time because its eggs were relatively bigger. The difference in reproductive rates is probably related to different partitioning of equal amounts of relative biomass: more small eggs for the aquatic rotifer vs. fewer big eggs for the terrestrial rotifer. Egg size might be related to the selective pressures of the environments.     

Cytological aspects of in vitro androgenesis in wheat (Triticum aestivum L.) using fluorescent microscopy

Summary Two winter wheat genotypes (Diószegi 200 and Mv 15) were compared for their in vitro androgenic capacity. On average, the induction frequency of embryogenic structures was 71.7% in Diószegi 200 and only 4.3% in Mv 15. The haploid induction ability of the two genotypes differed considerably, with Diószegi 200 being much higher. The difference in the in vitro inductability of the microspores may result from genetic differences which are manifested in the survival rate of the microspores during the culture period and their adaptability to in vitro conditions. Special DNA fluorochrornes were suitable for studying the different pathways of in vitro androgenesis. Our data indicate that the repeated equal divisions of the microspore nucleus might lead to pollen embryo formation, and subsequent divisions of the vegetative portion of the pollen grain after the first asymmetric microspore mitosis can result in pollen callus formation.