Mortality of American Bullfrog Tadpoles Lithobates catesbeianus Infected by Gyrodactylus jennyae and Experimentally Exposed to Batrachochytrium dendrobatidis

Abstract

The fungal pathogen Batrachochytrium dendrobatidis (Bd), which causes the disease chytridiomycosis in postmetamorphic amphibians, has been linked to amphibian population declines. Different amphibian species, however, exhibit different susceptibility to Bd pathogenicity. At the same time, agricultural pesticides commonly found contaminating aquatic habitats have been reported to increase the susceptibility of amphibians to pathogens. To investigate whether certain pesticides are able to alter the pathogenicity of Bd to larval amphibians, we exposed larval American bullfrogs Lithobates catesbeianus to end-use formulations of the herbicides atrazine or glyphosate, and then exposed them to Bd. Following the experimental exposures, a preexisting infection of the tadpoles by the monogenean ectoparasite Gyrodactylus jennyae was detected in all experimental and control tadpoles. Gyrodactylus jennyae infection intensity varied, and individuals with heavy G. jennyae infections suffered more skin erosion due to grazing by the parasite. Tadpoles experimentally exposed to Bd, or to Bd and either herbicide, had significantly reduced survival rates compared with untreated tadpoles that were only infected by G. jennyae. Increased mortality was also correlated with degree of skin erosion; survival of tadpoles with severe skin erosion was significantly reduced compared with that of tadpoles with no, or mild, skin erosion. While infected with G. jennyae, the group of tadpoles with the lowest survival rate (exposed only to Bd) included significantly more individuals exhibiting severe skin erosion and significantly fewer individuals without skin erosion, compared with the control group. These results emphasize the potential pathogenicity of gyrodactylid infections in larval amphibian hosts and suggest that concomitant exposures to Bd may enhance infections and effects of G. jennyae in bullfrog tadpoles.

Received February 3, 2012; accepted August 10, 2012     

Detecting and quantifying the adventitious presence of transgenic seeds in safflower, Carthamus tinctorius L

Safflower ( Carthamus tinctorius L.) is currently being developed as a platform for the production of novel proteins. Methods for detecting and quantifying transgenic safflower are needed to ensure seed quality and to monitor for its adventitious presence. We developed and compared three methods of assaying for transgenic safflower presence in conventional seedlots: field bioassays, enzyme-linked immunosorbent assays (ELISA), and quantitative polymerase chain reaction (Q-PCR). Limits for reliable quantification for both ELISA and Q-PCR are approximately 0.1%, although levels at least as low as 0.02% can be detected by Q-PCR. Levels of quantification for the field bioassay are limited only by space and resources available. Multiple sampling methods to detect and quantify transgenic safflower presence at levels lower than 0.1% were used on field collected samples from a pollen outcrossing experiment to quantify the adventitious presence of transgenic safflower. Taking into account the potential utility and relative advantages or disadvantages of each detection method, it is recommended that the initial testing for the adventitious presence of transgenic seed be carried out using an antibody-based test if available and that Q-PCR-based assays to quantify transgenic proportion be used when it is necessary to identify specific transgenic constructs or if antibody-based assays are not readily available.     

Discovery of lunasin peptide in triticale (X Triticosecale Wittmack)

Lunasin is a novel, cancer-preventive, anti-inflammatory and cholesterol-reducing peptide that was originally isolated from soy and later from barley, wheat and rye. We report the first discovery of lunasin in triticale (X Triticosecale Wittmack). Moreover, we report first data of lunasin content in winter rye and wheat genotypes grown in Northern Europe. These data are novel as previously published data on finding of lunasin in cereals were obtained in genotypes grown in Korea. Lunasin content was uncovered using a previously published procedure for isolation from cereals and identified by LC-MS/MS assay. We found that triticale was the most lunasin-rich cereal, with the tested genotypes displaying the following trend in lunasin content: genotype 0002-26 > Dinaro > DSGU 10/94 > 0213-22 > 0317-14 > 0006-31. The greatest lunasin content was 6.46 mg/g in the grain of triticale genotype 0002-26. In comparison, the highest lunasin content in rye variety Dankovske Diament was 1.5 mg/g, and the highest lunasin content in the winter wheat variety Fredis was 0.23 mg/g. We conclude that triticale can play a significant role as functional food, with great potential for the use of triticale products in human and animal diets.     

Discovery and directed evolution of a glyphosate tolerance gene

The herbicide glyphosate is effectively detoxified by N-acetylation. We screened a collection of microbial isolates and discovered enzymes exhibiting glyphosate N-acetyltransferase (GAT) activity. Kinetic properties of the discovered enzymes were insufficient to confer glyphosate tolerance to transgenic organisms. Eleven iterations of DNA shuffling improved enzyme efficiency by nearly four orders of magnitude from 0.87 mM-1 min-1 to 8320 mM-1 min-1. From the fifth iteration and beyond, GAT enzymes conferred increasing glyphosate tolerance to Escherichia coli, Arabidopsis, tobacco, and maize. Glyphosate acetylation provides an alternative strategy for supporting glyphosate use on crops.     

Identifying and measuring agrarian sentiment in regional Australia

In common with much of the Western world, agrarianism—valuing farmers and agricultural activity as intrinsically worthwhile, noble, and contributing to the strength of the national character—runs through Australian culture and politics. Agrarian sentiments and attitudes have been identified through empirical research and by inference from analysis of political debate, policy content, and studies of media and popular culture. Empirical studies have, however, been largely confined to the US, with little in the way of recent re-evaluations of, or developments from, early work. This paper reports on research that seeks quantitative empirical evidence for the existence of agrarianism in the Australian community and seeks to identify its core characteristics. Using a purpose-designed sub-set of items within a large, omnibus-style survey of regional and rural Australia, we demonstrate that agrarianism exists as a scientifically quantifiable concept identifiable through responses to four key propositions: that Australians should support policies aimed at improving the position of the agricultural industries; that working in agriculture and associated industries brings out the best in people; that agricultural producers make a major contribution to environmental protection and biodiversity conservation; and that the development of agriculture in Australia contributed to the development of the national character. We found very little variation in the degree to which different demographic groupings agree with agrarianism. Older people, farmers, and non-Indigenous Australian-born respondents were among those who were statistically significantly more likely to agree with the defining propositions of agrarianism, but their scores were only very slightly higher than those of other sub-populations in the sample.     

Signatures of adaptation to obligate biotrophy in the Hyaloperonospora arabidopsidis genome

Many oomycete and fungal plant pathogens are obligate biotrophs, which extract nutrients only from living plant tissue and cannot grow apart from their hosts. Although these pathogens cause substantial crop losses, little is known about the molecular basis or evolution of obligate biotrophy. Here, we report the genome sequence of the oomycete Hyaloperonospora arabidopsidis (Hpa), an obligate biotroph and natural pathogen of Arabidopsis thaliana. In comparison with genomes of related, hemibiotrophic Phytophthora species, the Hpa genome exhibits dramatic reductions in genes encoding (i) RXLR effectors and other secreted pathogenicity proteins, (ii) enzymes for assimilation of inorganic nitrogen and sulfur, and (iii) proteins associated with zoospore formation and motility. These attributes comprise a genomic signature of evolution toward obligate biotrophy.     

Outbreak of human infections with uncommon Salmonella serotypes linked to pet bearded dragons, 2012–2014

Reptiles are one of the fastest growing sectors in the United States pet industry. Reptile‐associated salmonellosis (RAS) continues to be an important public health problem, especially among children. We investigated an outbreak of human Salmonella infections resulting from serotypes Cotham and Kisarawe, predominately occurring among children. An outbreak of illnesses was identified in persons with exposure to pet bearded dragon lizards. Human and animal health officials, in cooperation with the pet industry, conducted epidemiologic, traceback and laboratory investigations. Onsite sampling was conducted at two US breeding facilities, one foreign breeding facility, and a large pet retail chain. A total of 166 patients in 36 states were identified with illness onset dates from 02/2012–06/2014. The median patient age was 3 years (range, <1–79 years), 57% were aged ≤5 years, and 37% were aged ≤1 year. Forty‐four patients (37%) were hospitalized, predominantly children. Sampling at breeding facilities and a national pet store chain resulted in isolation of outbreak serotypes at each facility; isolation proportions ranged from 2%–24% of samples collected at each facility.Epidemiologic, microbiologic and traceback evidence linked an outbreak of uncommon Salmonella serotypes to contact with pet bearded dragons. The high proportion of infants involved in this outbreak highlights the need to educate owners about the risk of RAS in children and the potential for household contamination by pet reptiles or their habitats. Strategies should be developed to improve breeding practices, biosecurity and monitoring protocols to reduce Salmonella in the pet reptile trade.