Genomic pedigree reconstruction identifies predictors of mating and reproductive success in an invasive vertebrate

The persistence of an invasive species is influenced by its reproductive ecology, and a successful control program must operate on this premise. However, the reproductive ecology of invasive species may be enigmatic due to factors that also limit their management, such as cryptic coloration and behavior. We explored the mating and reproductive ecology of the invasive Brown Treesnake (BTS: Boiga irregularis) by reconstructing a multigenerational genomic pedigree based on 654 single nucleotide polymorphisms for a geographically closed population established in 2004 on Guam (N = 426). The pedigree allowed annual estimates of individual mating and reproductive success to be inferred for snakes in the study population over a 14‐year period. We then employed generalized linear mixed models to gauge how well phenotypic and genomic data could predict sex‐specific annual mating and reproductive success. Average snout–vent length (SVL), average body condition index (BCI), and trappability were significantly related to annual mating success for males, with average SVL also related to annual mating success for females. Male and female annual reproductive success was positively affected by SVL, BCI, and trappability. Surprisingly, the degree to which individuals were inbred had no effect on annual mating or reproductive success. When juxtaposed with current control methods, these results indicate that baited traps, a common interdiction tool, may target fecund BTS in some regards but not others. Our study emphasizes the importance of reproductive ecology as a focus for improving BTS control and promotes genomic pedigree reconstruction for such an endeavor in this invasive species and others.     

Multiple paternity is a shared reproductive strategy in the live‐bearing surfperches (Embiotocidae) that may be associated with female fitness

According to Bateman's principle, female fecundity is limited relative to males, setting the expectation that males should be promiscuous, while females should be choosy and select fewer mates. However, several surfperches (Embiotocidae) exhibit multiple paternity within broods indicating that females mate with multiple males throughout the mating season. Previous studies found no correlation between mating success and reproductive success (i.e., a Bateman gradient). However, by including samples from a broader range of reproductive size classes, we found evidence of a Bateman gradient in two surfperch species from distinct embiotocid clades. Using microsatellite analyses, we found that 100% of the spotfin surfperch families sampled exhibit multiple paternity (Hyperprosopon anale, the basal taxon from the only clade that has not previously been investigated) indicating that this tactic is a shared reproductive strategy among surfperches. Further, we detected evidence for a Bateman gradient in H. anale; however, this result was not significant after correction for biases. Similarly, we found evidence for multiple paternity in 83% of the shiner surfperch families (Cymatogaster aggregata) sampled. When we combine these data with a previous study on the same species, representing a larger range of reproductive size classes and associated brood sizes, we detect a Bateman gradient in shiner surfperch for the first time that remains significant after several conservative tests for bias correction. These results indicate that sexual selection is likely complex in this system, with the potential for conflicting optima between sexes, and imply a positive shift in fertility (i.e., increasing number) and reproductive tactic with respect to the mating system and number of sires throughout the reproductive life history of females. We argue that the complex reproductive natural history of surfperches is characterized by several traits that may be associated with cryptic female choice, including protracted oogenesis, uterine sac complexity, and sperm storage.     

Genetic integrity of black sea bream (Acanthopagrus schlegeli) sperm following cryopreservation

Although semen cryopreservation has been applied successfully in many fish species, extensive variation in post‐thaw semen quality exists between species and individuals. AFLP (amplified restriction fragment length polymorphism) is a powerful method for detecting DNA polymorphisms at the individual, population, and species levels. The method has been successfully applied to boars (Sus domestica, Suidae, Artiodactyla, Mammalia) to detect and evaluate differences in DNA sequences that correspond with semen integretiy after employing various freezing techniques. Freezing and thawing of semen has also an effect of selecting for freezing‐resistant (or intact) and eliminating non‐viable or defective sperm. Only the fully intact and functional sperm, despite potential compromise by adverse freezing and osmotic stresses, retain fertility after thawing. The objective of this study was to use AFLP to assess any genetic changes associated with the effect of employed cryo‐methodology on the genetic integrity of sperm of the black sea bream (Acanthopagrus schlegeli) under different cryopreservation treatments. The cryopreservation protocols had no significant effect on sperm motility or survival rate of fertilized ova regardless of using fresh (% motile sperm 89.6 ± 3.0; % embryonic survival rate 54.4 ± 2.9) and frozen‐thawed semen (% motile sperm 80.2 ± 2.0; % embryonic survival rate 51.8 ± 2.0). The post‐thaw sperm motility and survival rates were not significantly different among the sperm samples of the five black sea bream males examined. In the present study, the remaining black sea bream sperm that survive the cryopreservation limit the power of AFLP to trace the genetic markers which correlate with the differences in the sensitivity of sperm to cryo‐injury. It is also possible that point mutations outside the AFLP priming sites may not have been detected. More thorough investigations are needed to determine whether such DNA fingerprints would be found in fish species.     

Direct genetic evidence for reproductive philopatry and associated fine‐scale migrations in female blacktip reef sharks (Carcharhinus melanopterus) in French Polynesia

Conservation of top predators has been emphasized as essential in an ecosystem due to their role in trophic chain regulation. Optimizing conservation strategies for these endangered marine top predators requires direct estimates of breeding patterns and connectivity as these are essential to understanding the population dynamics. There have been some attempts to investigate breeding patterns of reef sharks from litter reconstruction using molecular analyses. However, direct fine‐scale migrations of female sharks for parturition as well as connectivity at a medium scale like between islands remain mostly unknown. We used microsatellite DNA markers and a likelihood‐based parentage analysis to determine breeding patterns of female blacktip reef sharks in Moorea (Society Islands, French Polynesia). Most females gave birth at their home island but some migrated to specific nursery areas outside the area they are attached to, sometimes going to another island 50 km away across deep ocean. Our analysis also revealed that females migrated to the same nursery for every birthing event. Many offspring showed a high level of inbreeding indicating an overall reduced population size, restricted movements and dispersal, or specific mating behaviour. Females represent the vectors that transport the genes at nursery grounds, and their fidelity should thus define reproductive units. As females seem to be philopatric, males could be the ones dispersing genes between populations. These results highlight the need to conserve coastal zones where female reef sharks seem to exhibit philopatry during the breeding season.     

Environmental and demographic drivers of male mating success vary across sequential reproductive episodes in a polygynous breeder

Ecological and social factors underpinning the inequality of male mating success in animal societies can be related to sex ratio, sexual conflict between breeders, effects of nonbreeders, resource dispersion, climatic conditions, and the various sequential stages of mating competition that constitute the sexual selection process. Here, we conducted an individual‐based study to investigate how local resource availability and demography interact with annual climate conditions to determine the degree of male mating inequality, and thus opportunity for sexual selection across two sequential reproductive episodes (harem and subsequent mate acquisition) in a naturally regulated (feral) horse population in Sable Island National Park Preserve, Canada. Using a 5‐year, spatially explicit, mark‐resight dataset and hierarchical mixed‐effects linear modeling, we evaluated the influence of adult sex ratio (ASR) on mating success and then tested for effects of freshwater availability, density, unpaired male abundance, and precipitation during each breeding season. Unpaired male abundance, freshwater availability, and ASR differed in their effects on male mating success according to year and selection episode. Opportunity for sexual selection in males associated with harem acquisition increased with ASR, and unpaired male abundance further explained weather‐related interannual variation after accounting for ASR. In contrast, once a harem was secured, ASR had little effect on male mating inequality in regard to acquiring additional females, while interannual variation in mating inequality increased with decreasing freshwater availability. Our findings show that local demography, resource availability, and weather effect opportunity for sexual selection in males differently depending on selection episode, and can attenuate or accentuate effects of ASR.     

Gross anatomical and histomorphological features of the Acanthopagrus schlegelii digestive tract (Bleeker 1854) Perciformes,Sparidae

Acanthopagrus schlegelii is an autochthonous teleost species concerning the remarkable economic importance and prevalent fish cultivated in China as well as in different nations of South-East Asia. Little is known about the digestive tract (DT) morphology of A. schlegelii. Therefore, anatomical and histomorphological aspects of A. schlegelii DT were examined by light and scanning electron microscope (SEM). Anatomically, DT of A. schlegelii was mainly formed of oesophagus, well-developed stomach and fingers like pyloric caeca (four in number), intestinal regions and rectum. Histomorphologically, oesophagus occurred as a shorter tube-like organ presenting a longitudinal folded mucosa that connects oropharynx cavity to stomach. Stomach was a muscular thick-wall organ that included three regions, thickly longitudinal folds were observed in the first (cardiac) and last (pyloric) regions, whereas the second (fundic) region showed folds in different directions. Long villi were observed within pyloric caeca and anterior intestine. Tunica muscularis appeared narrow in the anterior intestine, whereas thicker in the posterior part of the intestine. Collectively, anatomical and histomorphological aspects of A. schlegelii DT are consistent with the carnivorous habit of this species. These data could be a potential source to modify better methods of nutrition and identify the DT pathogenic conditions in farming of this fish.     

Consequences of mating with siblings and nonsiblings on the reproductive success in a leaf beetle

Choosing a suitable mating partner is crucial for the fitness of an individual, whereby mating with siblings often results in inbreeding depression. We studied consequences of mating with siblings versus nonsiblings in the mustard leaf beetle, Phaedon cochleariae (Coleoptera: Chrysomelidae), on lifetime reproductive traits. Furthermore, we analyzed whether cuticular hydrocarbon (CHC) profiles are family specific and could potentially influence the mating behavior of young adults. We hypothesized a reduced reproductive success of females mated with siblings and a more rapid mating of males with nonsiblings. The hatching rate from eggs of sibling pairs was lower compared to that of nonsibling pairs, pointing to inbreeding depression. Furthermore, the number of eggs laid by females decreased over time in both sibling and nonsibling pairs. Interestingly, the CHC profiles and the body mass differed between families. However, the beetles did not avoid siblings and accepted them as readily as nonsiblings for mating in no‐choice tests. In summary, although it had negative consequences to mate a sibling and although siblings could potentially be recognized by their CHC profiles, the beetles did not show a delayed mating with siblings. Our results indicate that P. cochleariae beetles have not developed a precopulatory mechanism to avoid inbreeding, at least under the test conditions applied here. We predict that instead a polyandrous mating system and/or postcopulatory mechanisms might have evolved in this species by which inbreeding costs can be reduced.     

New records of a rare species of spariform fish Acanthopagrus schlegelii (Bleeker, 1854) (Osteichthyes: Sparidae) from Russian waters of the sea of Japan, with notes on the taxonomy of the genus Acanthopagrus Peters, 1855

A rare species of the spariform fish Acanthopagrus schlegelii from Russian waters is described. The synonymy of this species and the related literature data are analyzed. It is suggested that two species occur in Russian waters that differ in body coloration: A. schlegelii and A. czerskii.     

Intensive genetic assessment of the mating system and reproductive success in a semi-closed population of the mottled sculpin,Cottus bairdi

Most genetic surveys of parentage in nature sample only a small fraction of the breeding population. Here we apply microsatellite markers to deduce the genetic mating system and assess the reproductive success of females and males in an extensively collected, semi-closed stream population of the mottled sculpin fish, Cottus bairdi. In this species, males guard nest rocks where females deposit the eggs for fertilization. The potential exists for both males and females to mate with multiple partners and for males to provide parental care to genetically unrelated offspring. Four hundred and fifty-five adults and subadults, as well as 1,259 offspring from 23 nests, were genotyped at five polymorphic microsatellite loci. Multilocus maternal genotypes, deduced via genetic analyses of embryos, were reconstructed for more than 90% of the analysed nests, thus allowing both male and female reproductive success to be estimated accurately. There was no genetic evidence for cuckoldry, but one nest probably represents a takeover event. Successful males spawned with a mean of 2.8 partners, whereas each female apparently deposited her entire clutch of eggs in a single nest (mean fecundity = 66 eggs/female). On average, genetically deduced sires and dams were captured 1.6 and 9.3 metres from their respective nests, indicating little movement by breeders during the spawning season. Based on a 'genetic mark-recapture' estimate, the total number of potentially breeding adults (c. 570) was an order-of-magnitude larger than genetically based estimates of the effective number of breeders (c. 54). In addition, significantly fewer eggs per female were deposited in single than in multidam nests. Not only were perceived high-quality males spawning with multiple partners, but they were receiving more eggs from each female.     

Phytase can reduce theneedfor monocalcium phosphate supplementation in soybean and rapeseed meal‐based diets of black sea bream,Acanthopagrus schlegelii (Bleeker, 1854)

A feeding trial was conducted to study the effect of partial replacement of dietary monocalcium phosphate (MCP) with phytase on growth performance, feed utilization and phosphorus discharge in black sea bream, Acanthopagrus schlegelii. In the feeding trial, the control diet (designated as P1.5) was prepared with 1.5% MCP but without phytase, and the three other diets (designated as PP1.0, PP0.5 and PP0, respectively) were supplemented with 1.0%, 0.5% and 0% MCP, respectively, along with 200 mg (400 U) phytase/kg diet in each. Each diet was tested in triplicate tanks and fish were fed twice daily to satiation. After an 8‐week feeding trial in indoor flow‐through cylindrical fibreglass tanks (25 fish per tank, initial body weight: 11.5 ± 0.12 g), fish fed with PP1.0 and PP0.5 had no significant change in weight gain (WG), specific growth rate (SGR), protein efficiency rate (PER) or feed conversion ratio (FCR) compared to the control (p > .05), whereas fish fed with PP0 showed a significantly lower growth performance in the above parameters (p < .05). The addition of phytase did not affect the body composition or muscle composition. The apparent digestibility coefficients (ADCs) of crude protein and phosphorus increased when fish were fed diets in which MCP was replaced by phytase. Phosphorus discharge was also significantly reduced in fish fed diets in which MCP was replaced by phytase (10.2 ± 0.50 to 8.01 ± 0.47 g/kg weight gain). The present study suggests that dietary MCP can be reduced when phytase is added to the black sea bream diet, with a maximum MCP reduction level of up to 1% when phytase is supplemented at 200 mg (400 U)/kg diet. Thus, phytase in the diet of black sea bream is economically and ecologically beneficial.     

Quantitative measures of sexual selection reveal no evidence for sex-role reversal in a sea spider with prolonged paternal care

Taxa in which males alone invest in postzygotic care of offspring are often considered good models for investigating the proffered relationships between sexual selection and mating systems. In the pycnogonid sea spider Pycnogonum stearnsi, males carry large egg masses on their bodies for several weeks, so this species is a plausible candidate for sex-role reversal (greater intensity of sexual selection on females than on males). Here, we couple a microsatellite-based assessment of the mating system in a natural population with formal quantitative measures of genetic fitness to investigate the direction of sexual selection in P. stearnsi. Both sexes proved to be highly polygamous and showed similar standardized variances in reproductive and mating successes. Moreover, the fertility (number of progeny) of males and females appeared to be equally and highly dependent on mate access, as shown by similar Bateman gradients for the two sexes. The absence of sex-role reversal in this population of P. stearnsi is probably attributable to the fact that males are not limited by brooding space but have evolved an ability to carry large numbers of progeny. Body length was not a good predictor of male mating or reproductive success, so the aim of future studies should be to determine what traits are the targets of sexual selection in this species.     

Gamete traits influence the variance in reproductive success, the intensity of sexual selection, and the outcome of sexual conflict among congeneric sea urchins

Sea-urchin species differ in susceptibility to sperm limitation and polyspermy, but the influences of gamete traits on reproductive variance, sexual selection, and sexual conflict are unknown. I compared male and female reproductive success of two congeners at natural densities in the sea. The eggs of the species occurring at higher densities, Strongylocentrotus purpuratus, require higher sperm concentrations for fertilization but are more resistant to polyspermy compared to S. franciscanus. Both species show high variance in male fertilization success at all densities and high variance in female success at low densities, but they differ in female variance at high densities, where only S. franciscanus shows high female variance. The intensity of sexual selection based on Bateman gradients is high in males of both species, variable in S. franciscanus females, and low in S. purpuratus females. Strongylocentrotus franciscanus females experience sexual selection at low densities and sexual conflict at high densities. Strongylocentrotus purpuratus may rarely experience sperm limitation and may have evolved to ameliorate sexual conflict. This reduces the variance in female fertilization, providing females with more control over fertilization. Sperm availability influences sexual selection directly by determining sperm-egg encounter probabilities and indirectly through selection on gamete traits that alter reproductive variances.     

Alternative reproductive tactics and reproductive success in male Carollia perspicillata (Seba's short‐tailed bat)

The use of alternative reproductive tactics (ARTs) is widespread in animals. Males of some species may change tactics depending on age, body condition and social environment. Many bat species are polygynous where a fraction of males only have access to fertile females. For polygynous bats, knowledge of the reproductive success of males using different ARTs is scarce, and it remains unclear how age of males is related to switching decisions between social statuses. We studied a large captive population of Carollia perspicillata, where males are either harem holders, bachelors or peripheral males. Using a multistate procedure, we modelled the age‐related switches in reproductive tactics and in survival probability. From the model, we calculated the reproductive success and the frequencies of males displaying different reproductive tactics. As in mammals, the switch between social statuses is often related to age, we predicted that the transition probability of bachelor and peripheral males to harem status would increase with age. We show, however, that social status transition towards a harem holding position was not related to age. Reproductive success changed with age and social status. Harem males had a significantly higher reproductive success than bachelor males except between a short period from 3.8 to 4.4 years of age where success was similar, and a significantly higher reproductive success than peripheral males between 2.6 and 4.4 years of age. Harem males showed a clear decrease in the probability of maintaining social status with age, which suggests that senescence reduces resource holding potential.     

Phylogeny of Gracilariaceae (Rhodophyta): evidence from plastid and mitochondrial nucleotide sequences

Gracilariaceae are mostly pantropical red algae and include ~230 species in seven genera. Infrafamilial classification of the group has long been based on reproductive characters, but previous phylogenies have shown that traditionally circumscribed groups are not monophyletic. We performed phylogenetic analyses using two plastid (universal plastid amplicon and rbcL) and one mitochondrial (cox1) loci from a greatly expanded number of taxa to better assess generic relationships and understand patterns of character distributions. Our analyses produce the most well‐supported phylogeny of the family to date, and indicate that key characteristics of spermatangia and cystocarp type do not delineate genera as commonly suggested. Our results further indicate that Hydropuntia is not monophyletic. Given their morphological overlap with closely related members of Gracilaria, we propose that Hydropuntia be synonymized with the former. Our results additionally expand the known ranges of several Gracilariaceae species to include Brazil. Lastly, we demonstrate that the recently described Gracilaria yoneshigueana should be synonymized as G. domingensis based on morphological and molecular characters. These results demonstrate the utility of DNA barcoding for understanding poorly known and fragmentary materials of cryptic red algae.     

Structure of the female reproductive system of the lac insect,Kerria chinensis (Sternorrhyncha,Coccoidea: Kerridae)

The ovaries of female lac insects, Kerria chinensis Mahd (Sternorrhyncha: Coccoidea: Kerridae), at the last nymphal stage are composed of several balloon‐like clusters of cystocytes with different sizes. Each cluster consists of several clusters of cystocytes arranging in rosette forms. At the adult stage, the pair of ovaries consists of about 600 ovarioles of the telotrophic‐meroistic type. An unusual feature when considering most scale insects is that the lateral oviducts are highly branched, each with a number of short ovarioles. Each ovariole is subdivided into an anterior trophic chamber (tropharium) containing six or seven large trophocytes and a posterior vitellarium harbouring one oocyte which is connected with the trophic chamber via a nutritive cord. No terminal filament is present. Late‐stage adult females show synchronized development of the ovarioles, while in undernourished females, a small proportion of ovarioles proceed to maturity.     

Genetic and morphological evidence for two species of Leucocarbo shag (Aves,Pelecaniformes, Phalacrocoracidae) from southern South Island of New Zealand

Leucocarbo shags are a species‐rich seabird clade exhibiting a southern circumpolar distribution. New Zealand's endemic Stewart Island shag, Leucocarbo chalconotus (G. R. Gray, 1845), comprises two regional groups (Otago and Foveaux Strait) that show consistent differences in relative frequencies between pied (black and white) and bronze (wholly dark) plumages, the extent and colour of facial carunculation, body size (based on postcranial morphometrics), and breeding season. Moreover, previous genetic research on modern and historical specimens utilizing mitochondrial DNA control‐region sequences has also shown that the Otago and Foveaux lineages may not be sister taxa; instead, in several analyses the Otago lineage is sister to the endemic Chatham Island shag, Leucocarbo onslowi (Forbes, 1893). We present new ancient DNA analyses of the type specimens for the Otago and Foveaux Strait lineages of L. chalconotus, including a phylogenetic reanalysis of the available ancient, historical, and modern control‐region sequence data for these lineages (including L. onslowi), and additional statistical analyses incorporating new morphometric characters. These analyses indicate that under the diagnosable species concept the two lineages of Stewart Island shag represent two separate species, which we now recognize as the Otago shag, L. chalconotus (G. R. Gray, 1845), and the Foveaux shag, Leucocarbo stewarti (Ogilvie‐Grant, 1898).     

Pigment composition and photoacclimation as keys to the ecological success of Gonyostomum semen (Raphidophyceae,Stramenopiles)

Aquatic habitats are usually structured by light attenuation with depth resulting in different microalgal communities, each one adapted to a certain light regime by their specific pigment composition. Several taxa contain pigments restricted to one phylogenetic group, making them useful as marker pigments in phytoplankton community studies. The nuisance and invasive freshwater microalga Gonyostomum semen (Raphidophyceae) is mainly found in brown water lakes with sharp vertical gradients in light intensity and color. However, its pigment composition and potential photoadaptations have not been comprehensively studied. We analyzed the photopigment composition of 12 genetically different strains of G. semen by high performance liquid chromatography after acclimation to different light conditions. We confirmed the pigments chl a, chl c1c2, diadinoxanthin, trans‐neoxanthin, cis‐neoxanthin, α and β carotene, which have already been reported for G. semen. In addition, we identified, for the first time, the pigments violaxan‐thin, zeaxanthin, and alloxanthin in this species. Alloxanthin has never been observed in raphidophytes before, suggesting differences in evolutionary plastid acquisition between freshwater lineages and the well‐described marine species. The amount of total chl a per cell generally decreased with increasing light intensity. In contrast, the increasing ratios of the prominent pigments diadinoxanthin and alloxanthin per chl a with light intensity suggest photoprotective functions. In addition, we found significant variation in cell‐specific pigment concentration among strains, grouped by lake of origin, which might correspond to genetic differences between strains and populations.     

Unique mating behavior,and reproductive biology of a simultaneous hermaphroditic marine flatworm (Platyhelminthes,Tricladida, Maricola)

Flatworms generally are simultaneous hermaphrodites that exhibit various kinds of mating behavior. Here we report on the mating behavior and reproductive biology of the planarian Paucumara falcata. We recognized three phases in its mating behavior: a courtship, copulation, and postcopulatory phase. During the last‐mentioned phase, the partners showed a unique and very characteristic behavior in which their bodies intertwined, forming a spiral. Histological study of partners in copula revealed that the sclerotic tip of the musculo‐parenchymatic organ pierces the body wall of the partner and then becomes lodged in its parenchyma, suggesting that this organ may act as an anchor, thus stabilizing the worms during copulation. Similar organs in other species of marine triclad may also perform a stabilizing role during copulation. During copulation in individuals of P. falcata, sperm transfer was reciprocal or only unilateral. Copulation duration ranged 13–35 min (average 20 ± 5 min), irrespective of whether the mating was successful (i.e., resulted in the production of fertile cocoons). The spiraling phase lasted on average 10 min; some worms did not show the postcopulatory spiraling phase during their mating behavior. After successful copulation, an individual worm produced 1–12 fertile cocoons over a period of 1–17 days; from a cocoon hatched either one young (in 70% of the cases), or two young worms.     

Host influence on mating behavior and spermatophore reception correlated with reproductive output and longevity of femaleEarias insulana (Boisduval) (Lepidoptera: Noctuidae)

When mating and oviposition behavior ofEarias insulana were studied in the absence or presence of host or by providing only host odor, in comparison to other treatments host absence caused a delayed and reduced reproductive activity with occasional impairment of egg viability. Presence of host or its odor resulted in a short premating period and a significant increase in mating frequency, number of spermatophores transferred, number of eggs oviposited, and proportion of eggs fertilized. Compared to host odor, host accessibility had a larger impact on insect behavior, indicating the importance of host cantact. Under conditions of host accessibility, there existed a significant and positive correlation between the number of spermatophores received by a female and both the number of eggs she oviposited and her longevity. This indicated a role for male-derived nutrients in the reproduction ofE. insulana.