In vitro and in vivo effects of chemotherapeutants on the oyster parasite, Perkinsus marinus

To investigate the potential of chemotherapeutants to control the oyster pathogen Perkinsus marinus, anticoccidial and antifungal compounds were tested in vitro on infected hemolymph and cultured P. marinus cells and in vivo on infected oysters. In addition, acute toxicity to oysters was determined for six anticoccidials. In vitro experiments with infected hemolymph consisted of 24 h exposure of 0.2 mL hemolymph aliquots to concentrations ranging from 100 mg/L to 0.01 mg/L of amphotericin-B, amprolium, arprinocid, cycloheximide, lasalocid, malachite green, monensin, sulfadimethoxine, and a potentiated sulfadimethoxine, followed by incubation in fluid thioglycollate medium (FTM) to determine prezoosporangia abundance. Lasalocid, malachite green, and amphotericin-B were the most effective compounds reducing prezoosporangia abundance, relative to the untreated control group, at concentrations as low as 10 mg/L. Cycloheximide, monensin, and to a lesser extent sulfadimethoxine, were also effective but only at the highest concentration tested (100 mg/L). In vitro experiments with cultured P. marinus consisted of 24 h exposure of 10&\sp5& cells to 100 mg/L, 10 mg/L, and 1 mg/L of amphotericin-B, and 100 mg/L of cimetidine, cycloheximide, fumagillin, 5-fluorocytosine, ketoconazole, lasalocid, and monensin, followed either by incubation in FTM to determine abundance and size of prezoosporangia, or by addition of Neutral Red to determine cell viability. Amphotericin-B, lasalocid, and monensin were effective in reducing prezoosporangia abundance, size, and/or cell viability. No effects of cycloheximide on cultured cells were apparent. Lasalocid, monensin, and malachite green, were toxic to oysters at concentrations below 10 mg/L. The 96-hr. LC50 for lasalocid was 0.59 mg/L. No median lethal dose was determined for monensin or malachite green, but oyster mortality resulted from exposures ranging from 1 mg/L to 10 mg/L of either compound. In three in vivo experiments, infected oysters were exposed to amprolium, arprinocid, cycloheximide, lasalocid, monensin, malachite green, potentiated sulfadimethoxine, and sulfadimethoxine at various concentrations. Only cycloheximide was effective in reducing P. marinus infections. After 15 days of exposure to 10 mg/L of cycloheximide, weighted prevalence significantly declined from 3.78 in untreated controls to 2.10 in treated oysters. Infections progressed after treatment was discontinued as indicated by an increase in weighted prevalence from 0.71 at the end of treatment to 1.31 one month later. (Abstract shortened by UMI.)

Association of Fungal Secondary Metabolism and Sclerotial Biology

Fungal secondary metabolism and morphological development have been shown to be intimately associated at the genetic level. Much of the literature has focused on the co-regulation of secondary metabolite production (e.g., sterigmatocystin and aflatoxin in Aspergillus nidulans and Aspergillus flavus, respectively) with conidiation or formation of sexual fruiting bodies. However, many of these genetic links also control sclerotial production. Sclerotia are resistant structures produced by a number of fungal genera. They also represent the principal source of primary inoculum for some phytopathogenic fungi. In nature, higher plants often concentrate secondary metabolites in reproductive structures as a means of defense against herbivores and insects. By analogy, fungi also sequester a number of secondary metabolites in sclerotia that act as a chemical defense system against fungivorous predators. These include antiinsectant compounds such as tetramic acids, indole diterpenoids, pyridones, and diketopiperazines. This chapter will focus on the molecular mechanisms governing production of secondary metabolites and the role they play in sclerotial development and fungal ecology, with particular emphasis on Aspergillus species. The global regulatory proteins VeA and LaeA, components of the velvet nuclear protein complex, serve as virulence factors and control both development and secondary metabolite production in many Aspergillus species. We will discuss a number of VeA- and LaeA-regulated secondary metabolic gene clusters in A. flavus that are postulated to be involved in sclerotial morphogenesis and chemical defense. The presence of multiple regulatory factors that control secondary metabolism and sclerotial formation suggests that fungi have evolved these complex regulatory mechanisms as a means to rapidly adapt chemical responses to protect sclerotia from predators, competitors and other environmental stressors.This article is made openly accessible in part by an award from the Northern Illinois University Libraries’ Open Access Publishing Fund

Observed connections of Arctic stratospheric ozone extremes to Northern Hemisphere surface climate

We present observational evidence for linkages between extreme Arctic stratospheric ozone anomalies in March and Northern Hemisphere tropospheric climate in spring (March–April). Springs characterized by low Arctic ozone anomalies in March are associated with a stronger, colder polar vortex and circulation anomalies consistent with the positive polarity of the Northern Annular Mode/North Atlantic Oscillation in March and April. The associated spring tropospheric circulation anomalies indicate a poleward shift of zonal winds at 500 hPa over the North Atlantic. Furthermore, correlations between March Arctic ozone and March–April surface temperatures reveal certain regions where a surprisingly large fraction of the interannual variability in spring surface temperatures is associated with interannual variability in ozone. We also find that years with low March Arctic ozone in the stratosphere display surface maximum daily temperatures in March–April that are colder than normal over southeastern Europe and southern Asia, but warmer than normal over northern Asia, adding to the warming from increasing well-mixed greenhouse gases in those locations. The results shown here do not establish causality, but nevertheless suggest that March stratospheric ozone is a useful indicator of spring averaged (March–April) tropospheric climate in certain Northern Hemispheric regions.National Science Foundation (U.S.) (AGS-1539972