Paedomorphosis in the Ezo salamander (Hynobius retardatus) rediscovered after almost 90 years

Although paedomorphosis is widespread across salamander families, only two species have ever been documented to exhibit paedomorphosis in Hynobiidae. One of these two exceptional species is Hynobius retardatus in which paedomorphosis was first reported in 1924, in specimens from Lake Kuttara in Hokkaido. This population became extinct after the last observation in 1932; since then, no paedomorphs of this species have been reported anywhere. Here, we report the rediscovery of paedomorphs of this species. Three paedomorph-like male salamanders were collected from a pond in the south Hokkaido in December 2020 and April 2021; in size, these specimens were similar to metamorphosed adults but they still displayed larval features such as external gills and a well-developed caudal fin. An artificial fertilization experiment demonstrated that they were sexually compatible with metamorphosed females, thus, confirming them to be paedomorphs. Future efforts to find additional paedomorphs in this and other populations are required to assess the prevalence of paedomorphosis in H. retardatus and to improve understanding of the ecology and evolution of paedomorphisis in Urodela.

Separating Paternal and Maternal Contributions to Thermal Transgenerational Plasticity

Climate change is rapidly altering the thermal environment in terrestrial and aquatic systems. Transgenerational thermal plasticity (TGP) – which occurs when the temperatures experienced by the parental generation prior to the fertilization of gametes results in a change in offspring reaction norms – may mitigate the effects of climate change. Although “maternal effects” have been widely studied, relatively little is known about TGP effects in vertebrates, particularly paternal contributions. We used artificial fertilization to cross sheepshead minnow (Cyprinodon variegatus) parents exposed to either low (26°C) or high (32°C) temperatures and measured growth rates of the offspring over the first 8 weeks of life at both low and high temperatures. A linear mixed effects model was employed to quantify the effects of maternal, paternal, and offspring temperatures on offspring growth and fecundity. We found that the offspring growth rate up to 63 days post-hatch was affected by both the temperature they experienced directly and parental temperatures prior to fertilization. Growth was lowest when neither parents’ temperature matched the offspring temperature, indicating a strong transgenerational effect. Notably, offspring growth was highest when all three (offspring, sire, and dam) temperatures matched [although the three-way interaction was found to be marginally non-significant (P = 0.155)], suggesting that TGP effects were additive across significant sire-offspring (P < 0.001) and dam-offspring interactions (P < 0.001). Transgenerational effects on fecundity (GSI) were suggestive for both maternal and paternal effects, but not significant. The finding that thermal TGP is contributed by both parents strongly suggests that it has an epigenetic basis.

Optimization of Sperm Management and Fertilization in Zebrafish (Danio rerio (Hamilton))

The aim of the present study was to investigate the spontaneous motility of spermatozoa and to optimize sperm collection, short-term sperm storage, and fertilization in zebrafish Danio rerio. The movement of spermatozoon in water was propagated along the flagellum at 16 s after sperm activation then damped from the end of the flagellum for 35 s and fully disappeared at 61 s after activation. For artificial fertilization, milt must be added to an immobilizing solution, which stops the movement of sperm and keeps the sperm motionless until fertilization. E400 and Kurokura as isotonic solutions were shown to be suitable extenders to store sperm for fertilization for 6 h. E400 stored sperm for 12 h at 0–2 °C. Sperm motility decreased only to 36% at 12 h post stripping for the E400 extender and to 19% for the Kurokura extender. To achieve an optimal level of fertilization and swim-up larvae rates, a test tube with a well-defined amount of 6,000,000 spermatozoa in E400 extender per 100 eggs and 100 µL of activation solution has proven to be more successful than using a Petri dish. The highest fertilization and swim-up larvae rates reached 80% and 40–60%, respectively, with milt stored for 1.5 h in the E400 extender at 0–2 °C.

Centriolar defects, centrin 1 alterations, and FISH studies in human spermatozoa of a male partner of a couple that produces aneuploid embryos in natural and artificial fertilization

Purpose To study the potential paternal contribution to aneuploidies in the man of a couple who obtained trisomic embryos with natural and assisted fertilization. Methods Semen analysis, immunofluorescence for localization of tubulin and centrin 1, transmission electron microscopy (TEM), and fluorescence in situ hybridization (FISH) analysis for chromosomes 18 and 9 were performed. Sperm of fertile men were used as controls. Results The percentages of sperm motility and normal forms were decreased. The percentages of sperm with tail reduced in dimension, headless tails, coiled tails, and altered head-tail junction were significantly higher (P < 0.01) in the patient than in controls, whereas the percentage of sperm with a normal centrin 1 localization (two spots in the centriolar area) was significantly reduced (P < 0.01) in the patient. Immunofluorescence with anti-tubulin antibody showed that in most of the patient’s sperm connecting pieces (83.00 ± 1.78%), two spots were present, indicating prominent proximal centriole/centriolar adjunct and evident distal centriole, whereas controls’ sperm displayed a single spot, indicating the proximal centriole. The percentage of sperm with two spots was significantly higher (P < 0.01) in the patient than in controls. TEM analysis showed that centriolar adjuncts of the patient’s sperm were significantly longer (721.80 ± 122.26 nm) than in controls’ sperm (310.00 ± 64.11 nm; P < 0.001). The aneuploidy frequencies of the patient’s sperm, detected by FISH analysis, were increased with respect to controls. Conclusion A paternal contribution to sperm aneuploidies cannot be excluded since the patient’s sperm showed altered morphology, immature centriolar adjunct, presence of evident distal centriole, scarce presence of centrin 1, and high aneuploidy frequency.

Inseminating dose for the artificial fertilization of Brycon amazonicus (Teleostei: Characidae)

Summary Knowledge of the sperm–oocyte ratio in fish fertilization serves as the basis for studies on artificial reproduction and gamete manipulation. The aim of this study was to determine the minimum insemination dose for Brycon amazonicus oocyte fertilization. Female and male gametes were used and tested with the following doses of spermatozoa oocyte–1 ml–1: 10,000, 20,000, 40,000, 60,000 and 80,000 (in triplicate). Fertilization rates were calculated and estimated from the regression equation by applying the segmented regression model ‘Linear Response Plateau’ to determine the appropriate proportion of gametes. Based on the equation Ŷ = 14.3415 + 0.0007836X, the fertilization rate increased up to 63.34% as it reached a plateau with a proportion of 62,524 spermatozoa oocyte–1 ml–1, which is the minimum insemination dose recommended for artificial insemination of the species.

SINTASAN DAN PERTUMBUHAN LARVA CACING NIPAH Namalycastis rhodochorde (POLYCHAETA: NEREIDIDAE) PADA BUDIDAYA DENGAN DUA SUMBER PAKAN BERBEDA

Nypa Palm Worm Namalycastis rhodohorde has economic potential as an export commodity. Condition current population continues to decline due to habitat degradation and overharvesting. Cultivation be a solution to overcome these problems. On the laboratory scale this species has been successfully cultivated, but still faces obstacles to high mortality in early developmental stages. Larvae obtained by artificial fertilization and maintained with the same stocking density and container as well as the same volume of maintenance media only distinguished by two treated feed. Survival and growth of larvae feed did not differ between treatment seaweed and fermented palm fronds. Different stocking densities suggested for further research related to survival and growth of the Nypa palm worm larvae to obtain the best data for mass production

Larval development of nypa palm worm Namalycastis rhodochorde (Polychaeta: Nereididae)

Junardi, Anggraeni T, Ridwan A, Yowono E. 2020. Larval development of nypa palm worm Namalycastis rhodochorde (Polychaeta: Nereididae). Nusantara Bioscience 12: 148-153. Namalycastis intensive culture is still facing problems in mass production due to limited information on reproduction, especially in fertilization and production of larvae. The present research was designated to find out optimum salinity for fertilization and the production of nypa palm worm larvae (Namalycastis rhodochorde). Gamete samples were collected using a capillary glass tube inserted into ventro-lateral part of a body segment of a mature worm. Artificial fertilization was done by mixing the sperms and oocytes in a fertilization dish containing sterilized seawater as the medium. The larval development was observed until the benthic phase larvae (3-setigers). Fertilization was performed in medium salinity of 7-21‰ and water temperature of 25-29°C. The cleavage and larva stage occurred respectively within 28.20 to 58.67 minutes and within 72 to 80 hours after fertilization. The fertilization and larval development of N. rhodochorde were highly influenced by the medium salinity and temperature.