Propagation method of saving valuable strains from a Mycobacterium liflandii infection in Western clawed frogs (Silurana tropicalis).

Mycobacterium liflandii has been responsible for an emerging infection reported in the international trade of Western clawed frogs (Silurana tropicalis). This study shows that this mycolactone-producing Mycobacterium (MPM) has expanded its distribution range to France. The results of this study suggest that the use of in vitro fertilization to maintain genetic lines could be a temporary solution for valuable S. tropicalis propagation.

Localization of microcystin-LR in medaka fish tissues after cyanotoxin gavage.

Microcystins (MCs) are toxic monocyclic heptapeptides produced by many cyanobacteria. Over 70 MCs have been successfully isolated and identified, of which MC-LR is the most commonly occurring toxin. Microcystins, especially MC-LR, cause toxic effects in mammals, birds and fish and are a recognized potent cause of environmental stress and pose a potential health hazard in aquatic ecosystems when heavy blooms of cyanobacteria appear. They also constitute a public health threat to people via drinking water and food chains. The concentrations of MC-LR can be very low, even in fish displaying severely disrupted tissues, which makes it essential to devise selective and sensitive histochemical methods for identifying and localizing MC-LR in target organs, such as liver and intestine. The aim of the study reported here was to analyze the presence of MC-LR in contaminated fish tissues using immunohistochemical methods. The present experiment involving subacute exposure confirmed our initial hypothesis that subacute and acute exposure to microcystin contamination can exacerbate physiological stress, induce sustained pathological damage, and affect the immune response in exposed medaka fish.

Mycobacterium szulgai infection in a captive population of African clawed frogs (Xenopus tropicalis).

A colony of captive Xenopus tropicalis became infected with Mycobacterium szulgai. Clinical signs, when observed, were lethargy, weight loss, and emaciation. Visceral granulomas were common findings at laparoscopy and necropsy. The diagnosis of mycobacteriosis was based on histologic appearance and Ziehl-Neelsen staining of tissues. The identification of M. szulgai organisms was based on comparison of the 16S rRNA gene sequence with several GenBank databases. There have been no reports of this mycobacterial species as the causative agent of naturally occurring disease in amphibians.

Microcystin-LR and embryo-larval development of medaka fish, Oryzias latipes. I. Effects on the digestive tract and associated systems.

The cyanobacterial hepatotoxin microcystin-LR (MC-LR) is a specific potent PP1 and PP2A protein phosphatase inhibitor. In view to obtain an integrated whole-body, understanding of the key target organs of MC-LR subsequent to embryonic exposure on the anatomy of medaka fish hatchlings, embryos at stage 19 were microinjected with a sublethal dose of MC-LR corresponding to 0.2 pg/vitellus. MC-LR-induced histo-pathological modifications of the alimentary system (i.e. digestive tract, pancreas, liver) were analysed in newly hatched embryos. Our data are indicative of an MC-LR-induced inhibition of both yolk sac resorption and gas concentrating swimbladder expansion. In contrast to control hatchlings, (i) no mucus-secreting cells in the oesophagus, (ii) a decreased folding of the stomach and intestine, (iii) a clear reduction in size of the exocrine pancreas associated with a destructuration of acinar units, and (iv) a strong decrease in the mass and size of the liver were observed in MC-LR treated embryos. Furthermore, as an indication of MC-LR-induced hepatic glycogen store depletion, unstained cytoplasmic areas present in control hatchling hepatocytes, were fully absent in all liver examined in treated embryos. Finally, as a general observation in MC-LR-treated embryos, our data clearly indicated terminal differentiation disorders in all organs associated with the digestive tract.

Effects of microcystin-LR on development of medaka fish embryos (Oryzias latipes).

Chronic and subchronic toxicity from exposure to microcystins, cyclic hepatotoxic heptapeptides from cyanobacteria, receives increasing attention as a public human health biohazard. So far, the effects of microcystin on fish have been studied mainly in adults, rather than during early life stages. Limitations of direct ambient exposure experiments to fish egg have resulted from the difficult access of microcystin through the egg chorion. Using a microinjection technology, we have introduced microcystin-LR (MC-LR) directly into one-cell stage embryos or into the vitellus of late neurula embryos (stage 19) or into the vitellus of stage 25 embryos of medaka (Oryzias latipes) at the onset of the liver anlage. Microinjection (100 pl; stage 1 or 2 nl; stage 19 or 25) of MC-LR resulted in a dose dependent mortality of embryos. Survival rates were reduced up to 90% with microcystin concentrations of 10 or 1 microg/ml (corresponding to 1-20 pg or 0.1-2 pg of toxin injected), injected either at stages 1, 19 or 25. Also, a dose dependent advanced embryonic hatching processing was observed; hatching being brought forward from 2 or 3 days compared to controls in most of the microcystin injected groups. In agreement with the known hepatotoxic effects of microcystin, injected embryos consistently displayed hepatobiliary abnormalities such as liver hypertrophy and hepatic hemorrhage, also evidenced in post-hatching juveniles. Thus, the methodology presented in this paper should be valuable tool to analyze the effects of toxins on the development of aquatic vertebrate embryos.

The axolotl (Ambystoma mexicanum), a neotenic amphibian, expresses functional thyroid hormone receptors.

Neotenic amphibians such as the axolotl (Ambystoma mexicanum) are often unable to undergo metamorphosis under natural conditions. It is thought that neoteny represents a deviation from the standard course of amphibian ontogeny, affecting the thyroid axis at different levels from the central nervous system to peripheral organs. Thyroid hormone receptors (TRs) that bind the thyroid hormone (TH) T(3) have been described in axolotl. However, the full sequences of TR were needed to better characterize the TH response and to be able to assess their functional capacity at the molecular level. We report that each of the alpha and beta axolotl TRs bind both DNA and TH, and they activate transcription in response to TH in a mammalian cell-based transient transfection assay. Moreover, both TRs are expressed in axolotl tissues. Interestingly, each TR gene generates alternatively spliced isoforms, harboring partial or total deletions of the ligand-binding domain, which are expressed in vivo. Further, we found that in the axolotl, TH regulates the expression of stromelysin 3 and collagenase 3, which are TH target genes in Xenopus. Taken together, these results suggest that axolotl TRs are functional and that the molecular basis of neoteny in the axolotl is not linked to a major defect in TH response in peripheral tissues.