Addressing Reproducibility in Cryopreservation,and Considerations Necessary for Commercialization and Community Development in Support of Genetic Resources of Aquatic Species

For the past six decades a repeated cycle of developing new cryopreservation protocols or simply reinventing them to counteract a lack of reproducibility has led to hundreds of published studies that have offered little to the establishment of a genetic resources community for aquatic species. This has hampered repository development and inhibited industrial application. Most protocols were developed without standardized approaches, leading to irreproducible studies and questionable or meaningless comparisons. Thus cryopreservation of germplasm in aquatic species would greatly benefit from strategies to facilitate reproducibility. Our objectives were to: (1) identify major sources of irreproducibility across research, small‐scale, repository, and commercial‐scale development levels; (2) provide recommendations to address reproducibility challenges; and (3) offer suggestions on how researchers can directly influence commercial development and application of cryopreservation research. Sources of irreproducibility include lack of standardized procedural approaches, lack of standardized terminology, and lack of reporting guidelines. To address these challenges, we propose implementation of standard operating procedures, support of stock centers and internet content for development of training programs, and strengthening of the role of scientific journals and reviewers in reducing the frequency of irreproducible outcomes. Reproducibility is the foundation for quality management programs and product reliability, and therefore, standardization is necessary to assure efficient transition to commercial‐scale application and repository development. Progress can only be possible through community‐based approaches focused on coalescence and consensus of disparate groups involved in aquatic species cryopreservation and management of genetic resources.     

A field study of the impact of different irrigation practices on herbicide leaching

Agricultural practices, such as subsurface drainage, irrigation and tillage, may significantly affect pesticide leaching and, consequently, the risk of groundwater contamination. The aim of the present study was to investigate the impact of different irrigation systems on herbicide leaching to shallow groundwater through direct monitoring at the field scale in northern Italy over a 3-year period. Concentrations of the herbicide terbuthylazine (TBA) and its metabolite desethylterbuthylazine (DES) were monitored on 10 farms cropped with maize and irrigated by sprinkler, basin and border systems. Considering the results grouped according to the different irrigation systems, the mean TBA and DES concentrations was lower than the arbitrary non-health based legal limit of 0.1 μg/L using sprinkler and border systems, while it was 0.19 and 0.30 μg/L respectively for TBA and DES using basin systems.However, since many factors other than the irrigation systems can contribute to pesticide leaching and in a field study it is impossible to discriminate between all the different variables, the concentrations of both compounds were simulated with and without irrigation using the model MACRO 5.1 in order to gain a deeper understanding of the role of irrigation on leaching. First, the groundwater table depth, which was measured daily in all fields, was used to calibrate the model and thus achieve a good soil hydrology calibration. To assess the performance of the model the root mean squared error (RMSE) was used. RMSE ranged from 0.2 to 0.5 m, showing that a satisfactory hydrology calibration was obtained. Afterward, the solutes were modelled and the results showed that under non-irrigated conditions, concentrations of both compounds would be very low. These findings validate the hypothesis that careful selection of agricultural practices, such as the type of irrigation, can reduce pesticide leaching.     

Prevalence of Leptospira interrogans antibodies in free-ranging Tayassu pecari of the Southern Pantanal, Brazil, an ecosystem where wildlife and cattle interact

We surveyed a wild population of white-lipped peccaries (Tayassu pecari) in the Brazilian Pantanal for evidence of Leptospira interrogans. Serum samples from 71 free-ranging T. pecari were obtained between 2003 and 2005 in the southern Pantanal of Mato Grosso do Sul state. We used microscopic microagglutination to test for antibodies against 14 L. interrogans serovars (antibody titers ≥1:100 were considered seropositive). Seventy percent of captured animals tested positive for leptospirosis antibodies. Antibodies against icterohaemorrhagiae and autumnalis serovars were the most prevalent. We used log-linear analyses to test for associations among seropositivity, age class, and sex of captured animals. Seropositivity was strongly associated with animal age class, but independent of sex. Forty-six percent of animals less than 2 years old, 63% of adults during peak reproductive years, and 100% of the oldest age class were seropositive. A nonparametric multivariate procedure (MRPP) showed that the composition of serovar antibody types changed with age, and ANOVA models demonstrated that antibody titers increased with age, suggesting long-term exposure to a greater number and variety (i.e., serovar types) of L. interrogans infections. This study presents the first quantitative survey of antibodies against L. interrogans serovars in a T. pecari population of the Pantanal. The high prevalence of leptospirosis antibodies in free-ranging white-lipped peccaries and the potential impacts on reproduction and population dynamics emphasize the need for further studies investigating the roles of Pantanal wildlife and livestock in the transmission and maintenance of L. interrogans in the environment.     

Susceptibility of Trypanosoma evansi to propolis extract in vitro and in experimentally infected rats

Current therapy of Trypanosoma evansi infections is not effective for the vast majority of animals with relapsing parasitemia and clinical signs. Recently, attention is being focused on the antiparasitic activity of propolis. This study evaluated the susceptibility of T. evansi to propolis extract in vitro and in vivo. A dose-dependent trypanocidal activity of propolis extract was observed in vitro. All trypomastigotes were killed 1h after incubation with 10μgmL(-1) of the extract. In vivo, the concentrations of 100, 200, 300 and 400mgkg(-1) administered orally for 10 consecutive days showed no curative effect, and the rats died from the disease. However, rats treated with the two highest concentrations of propolis extract showed higher longevity than the other groups. Based on these data, we concluded that T. evansi is susceptible to propolis in vitro. Despite the lack of curative efficacy observed in vivo at the concentrations tested, the propolis extract can prolong life in rats infected with the protozoan.     

Exploiting multimedia in reproductive science education: research findings

Education in reproductive science is operating from an outdated paradigm of teaching and learning. Traditionally, reproductive education follows the pattern where students read a textbook, listen to instructor presentations, re-read the textbook and class notes and then complete a test. This paradigm is inefficient, costly and has not incorporated the potential that technology can offer with respect to increases in student learning. Further, teachers of reproductive science (and all of science for that matter) have little training in the use of documented methods of instructional design and cognitive psychology. Thus, most of us have learned to teach by repeating the approaches our mentors used (both good and bad). The technology now exists to explain complex topics using multimedia presentations in which digital animation and three-dimensional anatomical reconstructions greatly reduce time required for delivery while at the same time improving student understanding. With funding from the Small Business Innovation Research program through the U.S. Department of Education, we have developed and tested a multimedia approach to teaching complex concepts in reproductive physiology. The results of five separate experiments involving 1058 university students and 122 patients in an OB/GYN clinic indicate that students and patients learned as much or more in less time when viewing the multimedia presentations when compared to traditional teaching methodologies.