In vivo toxicity of bioreactor-grown biomass and exopolysaccharides from Malaysian tiger milk mushroom mycelium for potential future health applications

Natural mycelial biomass (MB) and exopolysaccharides (EPS) of Malaysian tiger milk mushroom Lignosus rhinocerus are considered high-end components due to their high commercial potential value in drug discovery. This study aims to evaluate the toxicity of the mushroom extracts’ generated in a bioreactor using the zebrafish embryo toxicity (ZFET) model assay as a new therapy for treating asthma. Both MB and EPS extracts, at concentrations 0.16–10 mg/mL, were tested for ZFET and early development effects on Zebrafish Embryos (ZE) during 24–120 h post-fertilisation (HPF). Findings revealed that MB was deemed safe with an LC50 of 0.77 mg/mL; the EPS were non-toxic (LC50 of 0.41 mg/mL). Neither MB nor EPS delayed hatching nor teratogenic defects in the treated ZE at a 2.5 mg/mL dose. There were no significant changes in the ZE heart rate after treatments with MB (130 beats/min) and EPS (140 beats/min), compared to that of normal ZE (120–180 beats/min). Mixing both natural compounds MB and EPS did not affect toxicity using ZFET testing; thus, intimating their safe future use as therapeutic interventions. This represents the first study to have used the ZFET assay on MB and EPS extracts of L. rhinocerus for future health applications.

Delayed mite hatching in response to mechanical stimuli simulating egg predation attempts

Delayed or induced hatching in response to predation risk has been reported mainly in aquatic systems, where waterborne cues from predators and injured neighbouring eggs are available. Newly emerged larvae of the terrestrial predatory mite Neoseiulus womersleyi are vulnerable to predation by con- and heterospecific predatory mites, whereas their eggs are not. We examined whether N. womersleyi embryos delay hatching in response to artificial mechanical stimuli that simulates egg predation attempts. When embryos near the hatching stage were artificially stimulated every 5 min for 60 min, most stopped hatching for the duration of the 60-min period, whereas unstimulated embryos did not. Stimulated embryos resumed hatching when the treatment was stopped, and the proportion of hatched stimulated embryos caught up with that of unstimulated embryos within 120 min after stimuli stopped. Since hatching did not stop in response to changes in gravity direction, the effect of direct mechanical stimuli on the eggs was considered a proximate factor in delayed hatching. These results suggest that N. womersleyi embryos recognise immediate predation risk via mechanical stimuli, and delay hatching until the predation risk is reduced.

Oviposition drives hatching order and developmental disparities with brood mates

Birth or hatching order can affect fitness. It has long been assumed that the order in which a bird's egg is laid (its oviposition) and first exposed to incubation relative to other eggs in the nest determines the order in which it hatches and the subsequent effects on development and survival. To my knowledge, this cause of hatching order has not been tested while controlling for laying-order effects on egg composition. Like those of many species, the last-laid eggs in clutches of the Lincoln's sparrow ( Melospiza lincolnii ) hatch last and produce nestlings with reduced growth and survival rates. Last-hatched Lincoln's sparrows also develop wider bills that are associated with reduced attractiveness of adult courtship songs. Using wild, free-ranging Lincoln's sparrows, I replaced the first or fourth freshly laid egg in four-egg host nests with a freshly laid first egg from a donor nest. I discovered that the experimental elevation of oviposition—which controlled for egg composition—delayed hatching, inhibited growth and widened bill gape, thus confirming the mechanism for hatching order and revealing a potential role for oviposition in reproductive success. Similar effects of postnatal family position could affect fitness in other taxa, as well.

Toxicity of 2 pg ethynylestradiol in brown trout embryos (Salmo trutta)

Endocrine disrupting chemicals are a threat to natural fish populations in the aquatic environment. Their toxicity is usually discussed relative to concentrations in the water the fish are exposed to. In the case of the synthetic compound 17-alpha-ethynylestradiol (EE2), a common and persistent estrogen, concentrations around 1 ng/L have repeatedly been found to induce toxic effects in fish. Here, we used brown trout (Salmo trutta) from a natural population to study EE2 take up and how it affects early life-history. We collected adults during the spawning season, produced 730 families in vitro (to control for potential maternal and paternal effects on embryo stress tolerance), and singly raised 7,300 embryos (in a 2 mL static system) that were either exposed to one dose of EE2 at 1 ng/L (i.e., 2 pg/embryo) or sham-treated. We found that EE2 concentration did not significantly change over a period of 3 months in control containers without embryos. Embryos took up most of the 2 pg EE2 within about 4 weeks at 4.6°C. EE2 treated embryos experienced higher mortality, delayed hatching of the survivors, and had reduced size at hatching. Our findings suggest that the toxicity of EE2 is often underestimated when discussed at the level of concentrations in water only.