In vertebrates, sex determination occurs along a continuum from strictly genotypic (GSD), where sex is entirely guided by genes, to strictly environmental (ESD), where rearing conditions, like temperature, determine phenotypic sex. Along this continuum are taxa which have combined genetic and environmental contributions to sex determination (GSD + EE), where some individuals experience environmental effects which cause them to sex reverse and develop their phenotypic sex opposite their genotypic sex. Amphibians are often assumed to be strictly GSD with sex reversal typically considered abnormal. Despite calls to understand the relative natural and anthropogenic causes of amphibian sex reversal, sex reversal has not been closely studied across populations of any wild amphibian, particularly in contrasting environmental conditions. Here, we use sex-linked molecular markers to discover sex reversal in wild populations of green frogs (Rana clamitans) inhabiting ponds in either undeveloped, forested landscapes or in suburban neighborhoods. Our work here begins to suggest that sex reversal may be common within and across green frog populations, occurring in 12 of 16 populations and with frequencies of 2–16% of individuals sampled within populations. Additionally, our results also suggest that intersex phenotypic males and sex reversal are not correlated with each other and are also not correlated with suburban land use. While sex reversal and intersex are often considered aberrant responses to human activities and associated pollution, we found no such associations here. Our data perhaps begin to suggest that, relative to what is often suggested, sex reversal may be a relatively natural process in amphibians. Future research should focus on assessing interactions between genes and the environment to understand the molecular and exogenous basis of sex determination in green frogs and in other amphibians.

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A field experiment was carried out in northern Vietnam to investigate the effects of adding different additives [rice (Oriza sativa L.) straw only, or rice straw with added lime, superphosphate (SSP), urea or a mixture of selected microorganism species] on nitrogen (N) losses and nutrient concentrations in manure composts. The composts and fresh manure were applied to a three-crop per year sequence (maize-rice-rice) on a degraded soil (Plinthic Acrisol/Plinthaquult) to investigate the effects of manure type on crop yield, N uptake and fertilizer value. Total N losses during composting with SSP were 20% of initial total N, while with other additives they were 30-35%. With SSP as a compost additive, 65-85% of the initial ammonium-N (NH 4 -N) in the manure remained in the compost compared with 25% for microorganisms and 30% for lime. Nitrogen uptake efficiency (NUE) of fresh manure was lower than that of composted manure when applied to maize (Zea mays L.), but higher when applied to rice (Oriza sativa L.). The NUE of compost with SSP was generally higher than that of compost with straw only and lime. The mineral fertilizer equivalent (MFE) of manure types for maize decreased in the order: manure composted with SSP > manure composted with straw only and fresh manure > manure composted with lime. For rice, the corresponding order was: fresh manure > manure composted with SSP/microorganisms/urea > manure composted with lime/with straw alone. The MFE was higher when 5 tons manure ha À1 were applied than when 10 tons manure ha À1 were applied throughout the crop sequence. The residual effect of composted manures (determined in a fourth crop, with no manure applied) was generally 50% higher than that of fresh manure after one year of manure and compost application. Thus, addition of SSP during composting improved the field fertilizer value of composted pig manure the most.