This paper highlights the teratogenic and toxic effects of Ganoderma lucidum (Lingzhi or Reishi mushroom) extract on zebrafish embryos. Hatchability, malformations, and lethality rate of zebrafish embryos were assessed to provide valuable information regarding the potential teratogenic activity of G. lucidum. Hatching was completed 48 h post treatment application (hpta) at 1% or lower concentrations of extract and embryo water. The hatching rate of embryos treated with 5% or higher concentrations was significantly lower (p> 0.05) than the control. Tail malformation was the most marked morphological abnormality in embryos at 72 hpta, which was obviously caused by 1% extract (55.56% tail malformation) and was observed in all embryos exposed to 5% of extract. Growth retardation was evident in embryos exposed to 5%, 10%, and 20%. However, lethal effect of extract of G. lucidum was dependent on dose and time of exposure. Mortality rates of embryos treated with 5% (44.44%) or higher concentrations of the extract was significantly higher (p > 0.05) than that of the control embryos at 72 hpta. These results suggest that G. lucidum extract has lethal and sub-lethal effects on zebrafish embryos.

The fertilization and cleavage of bovine oocytes matured by intra- or extra-follicular methods were investigated. Oocytes were fertilized in vitro or in the rabbit oviduct and cleavage was assessed after in vitro culture of in vitro fertilized oocytes and after in vivo culture (rabbit oviducts) of xenogenously fertilized oocytes. The effect of fertilization with fresh-diluted or frozen-thawed semen were also examined. The intra-follicular method did not increase the nuclear maturation rate as compared with the extra-follicular method (57.9 and 52.7%, respectively). However, the proportions of in vitro fertilized eggs (54.8%) and of cleaved eggs (two to eight cells; 34.6%) in the rabbit oviduct for 48 h after xenogenous fertilization were higher (P less than .025) in the intra-follicular oocytes than those of the extra-follicular oocytes (37.1 and 21.3%, respectively). It was also found that the use of fresh-diluted semen resulted in more cleaved eggs from the rabbit oviduct than the use of frozen-thawed semen (43.4 and 23.3% in the intra-follicular oocytes, P less than .025; 31.0 and 7.8% in the extra-follicular oocytes, P less than .05), while the appearance of cleaved eggs following in vitro fertilization was extremely low (0 to 6.6%). The present results demonstrated that the intra-follicular culture method of bovine oocytes provided a physiological environment for cytoplasmic maturation leading to higher fertilizability and development than the conventional in vitro culture of extra-follicular oocytes.

Arius manillensis is an endemic and economically important catfish of Laguna de Bay, Philippines. This study generated data on some aspects of its reproductive biology that are vital for the enhancement of propagation efforts for inland fisheries in the Southeast Asian region. Monthly assessments were conducted for one whole year to establish the distinct spawning season(s) through determinations of gonadosomatic index (GSI), fecundity through counting the number of two stages of follicles, and size at sexual maturity. Sex ratio was taken into account. Peaks in GSI occurred in March to April and July to August. These peaks correspond with months of peaks in fecundity. The parallel patterns of these reproductive aspects strengthen the occurrence of two distinct spawning seasons. GSI did not show significant correlation with body parameters while fecundity was significantly correlated with body weight (BW) and length (BL) and gonad weight (GW). Size and weight of apparent sexual maturity or peak reproductive capacity is about 20 cm and 200 g, respectively. Sex ratio was female-biased (2.11: 1). Female dominance indicates a unique reproductive behavior of A. manilensis where males could have evolved survival mechanism(s) to elude capture during breeding seasons and evidently during spawning on the cold months.

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