Validation of the indirect fluorescent antibody and the complement fixation tests for the diagnosis of Theileria equi

The indirect fluorescent antibody (IFA) test for Theileria equi was evaluated to assess test's suitability for the serological diagnosis of equine piroplasmosis, to provide performance parameters for the purpose of test validation, and to compare it with the complement fixation (CF) test. Using a protocol that included Evan's blue, the specificity of the IFA test was estimated at 99.0% for T. equi by the classical method of analysis, and 96.6% by the Bayesian method. The use of Evan's blue in the test protocol increased test specificity and contributed to an excellent test agreement between two collaborating laboratories (kappa = 0.96). Using Bayesian analysis, the sensitivity estimate for the IFA test was 89.2%. The CF test sensitivity and specificity estimates for T. equi were 63.1 and 96.4%, respectively, as determined by Bayesian analysis. The IFA test was more sensitive than the CF test but the specificity estimates were similar.     

Environmental Manipulation of Growth and Sexual Maturation in Yellow Perch, Perca flavescens

Yellow perch, Perca flavescens, exhibit sexually dimorphic growth detected as the fish enter puberty. The purpose of this study was to evaluate the differential influences of photoperiod and temperature on the sexual maturation and dimorphic growth pattern of yellow perch. Four combinations of constant (CP) or natural (NP) photoperiod with constant (CT) or natural (NT) temperature were evaluated (CPCT, CPNT, NPCT, NPNT). Following 6 months of growth, results indicate that (1) CT is the strongest promoter of overall growth regardless of photoperiod. Perch reared under CT were 26.4 ± 8.1 g heavier than perch reared under NT (P < 0.0001). (2) CP also promotes overall growth regardless of temperature. CP perch outgrew NP perch by 7.0 ± 8.1 g (P < 0.0872). (3) NP (decreasing) initiates maturation in yellow perch regardless of temperature. (4) NT is required for late maturational processes. While both NP groups initiated maturation, only NPNT females were able to recruit oocytes to a vitellogenic stage. Only NPNT males were able to sustain spermiation to the end of the study. (5) CPCT confer the best overall growth performance, fully inhibit maturation in both genders, and suppress a sexually dimorphic growth pattern up to 53 g average weight. These results indicate that using a CPCT regime during intensive fingerling production will produce a larger, more uniform fish population that could alleviate losses as a result of cannibalism. Perch that remained immature did not exhibit a dimorphic growth pattern.     

Novel Prunus rootstock somaclonal variants with divergent ability to tolerate waterlogging

Plants require access to free water for nutrient uptake, but excess water surrounding the roots can be injurious or even lethal because it blocks the transfer of free oxygen between the soil and the atmosphere. Genetic improvement efforts in this study were focused on the increased tolerance in roots to waterlogging. Among a pool of clones generated in vitro from leaf explants of rootstock Mr.S.2/5 of Prunus cerasifera L., the S.4 clone was flood tolerant whereas the S.1 clone was sensitive. The S.4 clone formed adventitious roots on exposure to flooding. Moreover, the chlorophyll content and mitochondrial activity in the leaf and root, soluble sugar content, alcohol dehydrogenase activity and ethylene content were different between the clones. The sorbitol transporter gene (SOT1) was up-regulated during hypoxia, the alcohol dehydrogenase genes (ADH1 and ADH3) were up-regulated in the leaves and down-regulated in the roots of the S.4 clone during hypoxia, and the 1-aminocyclopropane-1-oxidase gene (ACO1) was up-regulated in the leaves and roots of the S.4 clone during hypoxia and down-regulated in the wild-type roots. In addition, in the S.4 root, hypoxia induced significant down-regulation of a glycosyltransferase-like gene (GTL), which has a yet-undefined role. Although the relevant variation in the S.4 genome has yet to be determined, genetic alteration clearly conferred a flooding-tolerant phenotype. The isolation of novel somaclonals with the same genomic background but with divergent tolerance to flooding may offer new insights in the elucidation of the genetic machinery of resistance to flooding and aid in the selection of new Prunus rootstocks to be used in various adverse environments.     

Systolic time intervals assessed by 2-D echocardiography and spectral Doppler in the horse

In the present study, the ‘acoustic windows’ for the measurement of the left ventricular systolic time intervals is studied by means of 2‐D echocardiography and cardiac Doppler ultrasonography, and the normal values in the horse (n = 112) are determined. The left ventricular isovolumetric contraction time, the pre‐ejection period (PEP), as well as the left ventricular ejection time (LVET) have been measured, and the values of the left ventricular total electromechanical systole (LVTES) and the PEP‐to‐LVET ratio have been calculated. It has been established that the most suitable window for the measurement of the aforementioned indices in 2‐D echocardiography is the right parasternal window in a view in short axis at the level of the cardiac base to measure the aortogram. In Doppler ultrasonography, the preferred window is the left parasternal using the five chambers apical view. The following values have been acquired: PEP = 0.071 ± 0.01 s; LVET = 0.532 ± 0.097 s; LVTES = 0.6 ± 0.1 s and PEP‐to‐LVET = 0.138 ± 0.025 measured by 2‐D echocardiography and PEP = 0.068 ± 0.009 s; LVET = 0.527 ± 0.076 s; LVTES = 0.598 ± 0.098 s and PEP‐to‐LVET = 0.131 ± 0.01 measured by Doppler ultrasonography.     

Anabolism,catabolism and proteomic analysis in the slow‐twitch muscle of pacu (Piaractus mesopotamicus) submitted to prolonged fasting followed by refeeding

Here, we aimed to study the slow muscle of the fish Piaractus mesopotamicus submitted to 30 days of fasting (D30) followed by 1 day (D31) or 30 days of refeeding (D60). Histological analysis of fibre diameter was performed in D30 and D60. The gene expression of parvalbumin (pvalb), atrogenes (murf1a, murf1b, mafbx) and anabolic genes (igf‐1, mtor) was analysed using RT‐qPCR in D30, D31 and D60. The proteome was obtained by shotgun proteomics at D30 and D60, and the set of differentially expressed proteins was analysed by bioinformatics. In all experiments, the control was regularly fed fish. The histological analysis showed no changes in muscle fibre diameter. The expression of catabolic and anabolic genes was not changed, except for the downregulation of igf‐1 in D30 and of mafbx in D31. The expression of pvalb was not changed in D30 and D60 but was decreased in D31. The proteomic analysis identified 169 proteins in D30 (24 upregulated and 18 downregulated) and 170 proteins in D60 (17 upregulated and 21 downregulated); many of them were related to energetic metabolism and intracellular Ca²⁺ homoeostasis. Overall, our results indicate that the slow‐twitch muscle presented few changes upon prolonged fasting and refeeding condition.     

Antiplatelet and Anticoagulant Effects of Diterpenes Isolated from the Marine Alga,Dictyota menstrualis

Cardiovascular diseases represent a major cause of disability and death worldwide. Therapeutics are available, but they often have unsatisfactory results and may produce side effects. Alternative treatments based on the use of natural products have been extensively investigated, because of their low toxicity and side effects. Marine organisms are prime candidates for such products, as they are sources of numerous and complex substances with ecological and pharmacological effects. In this work, we investigated, through in vitro experiments, the effects of three diterpenes (pachydictyol A, isopachydictyol A and dichotomanol) from the Brazilian marine alga, Dictyota menstrualis, on platelet aggregation and plasma coagulation. Results showed that dichotomanol inhibited ADP- or collagen-induced aggregation of platelet-rich plasma (PRP), but failed to inhibit washed platelets (WP). In contrast, pachydictyol A and isopachydictyol A failed to inhibit the aggregation of PRP, but inhibited WP aggregation induced by collagen or thrombin. These diterpenes also inhibited coagulation analyzed by the prothrombin time and activated partial thromboplastin time and on commercial fibrinogen. Moreover, diterpenes inhibited the catalytic activity of thrombin. Theoretical studies using the Osiris Property Explorer software showed that diterpenes have low theoretical toxicity profiles and a drug-score similar to commercial anticoagulant drugs. In conclusion, these diterpenes are promising candidates for use in anticoagulant therapy, and this study also highlights the biotechnological potential of oceans and the importance of bioprospecting to develop medicines.     

Quantification and persistence of recombinant DNA of Roundup Ready corn and soybean in rotation

The presence of the recombinant cp4 epsps gene from Roundup Ready (RR) corn and RR soybean was quantified using real-time PCR in soil samples from a field experiment growing RR and conventional corn and soybean in rotation. RR corn and RR soybean cp4 epsps persisted in soil for up to 1 year after seeding. The concentration of recombinant DNA in soil peaked in July and August in RR corn and RR soybean plots, respectively. A small fraction of soil samples from plots seeded with conventional crops contained recombinant DNA, suggesting transgene dispersal by means of natural process or agricultural practices. This research will aid in the understanding of the persistence of recombinant DNA in agricultural cropping systems.     

Physiological and biochemical characterization of quinclorac resistance in a false cleavers (Galium spurium L.) biotype

The physiological and biochemical basis for quinclorac resistance in a false cleavers (Galium spurium L.) biotype was investigated. There was no difference between herbicide resistant (R) and susceptible (S) false cleavers biotypes in response to 2,4-D, clopyralid, glyphosate, glufosinate-ammonium, or bentazon. On the basis of GR(50) (growth reduction of 50%) or LD(50) (lethal dose to 50% of tested plants) values, the R biotype was highly resistant to the acetolactate synthase (ALS) inhibitor, thifensulfuron-methyl (GR(50) resistance ratio R/S = 57), and quinolinecarboxylic acids (quinclorac R/S = 46), resistant to MCPA (R/S = 12), and moderately resistant to the auxinic herbicides picloram (R/S = 3), dicamba (R/S = 3), fluroxypyr (R/S = 3), and triclopyr (R/S = 2). The mechanism of quinclorac resistance was not due to differences in [(14)C]quinclorac absorption, translocation, root exudation, or metabolism. Seventy-two hours after root application of quinclorac, ethylene increased ca. 3-fold in S but not R plants when compared to controls, while ABA increased ca. 14-fold in S as opposed to ca. 3-fold in R plants suggesting an alteration in the auxin signal transduction pathway, or altered target site causes resistance in false cleavers. The R false cleavers biotype may be an excellent model system to further examine the auxin signal transduction pathway and the mechanism of quinclorac and auxinic herbicide action.     

Trapping for mink control and water vole survival: Identifying key criteria using a spatially explicit individual based model

Introduced alien species are the second most important threat to global biodiversity after habitat loss. The American mink Mustela vison has been introduced to several countries and is threatening a number of native species worldwide. We developed a spatially explicit and individual based model as a planning tool to identify key criteria for the implementation of trapping campaigns as a way to control open American mink populations. We first predicted the minimum effort required to reduce populations of mink below a certain threshold and the best time of year in which to trap mink to minimise their numbers. We then employed this methodology to predict the best trapping strategy to ensure the long-term survival of the water vole Arvicola terrestris, one of the species most endangered by the spread of the mink in the UK. We also applied the mink and water vole population models to rationalise a set of observed data in an area of 50 × 30 km in the Upper Thames (UK). The model predicted that it is necessary to remove mink for at least 3 months every year and that a mixed strategy of trapping during the mating, late dispersal and winter seasons is best for keeping mink at low densities. Concentrating trapping during the late dispersal and winter seasons is instead best for ensuring the long-term survival of water voles. Targeting immigrating juvenile mink as well as reproductive adults is important. The model also showed that trapping efficiency might be an important factor to consider when choosing periods in which to trap.