Molecular characterization of potentially zoonotic isolates of Giardia duodenalis in horses

Giardia isolates from eight horses from New York State (NY), USA and two horses from Western Australia (WA) were genetically characterized at the SSU-rDNA and triose-phosphate isomerase (TPI) genes. Phylogenetic analysis of the TPI gene provided strong support for the placement of both isolates of Giardia from horses in WA and a single isolate from a horse in NY within the assemblage AI genotype of G. duodenalis. Another two isolates from horses in NY placed within the assemblage AII genotype of G. duodenalis. Phylogenetic analysis of the TPI gene also provided strong bootstrap support for the placement of four G. duodenalis isolates from horses in NY into a potentially host-specific sub-assemblage of assemblage BIV. The results of this study are consistent with previous studies showing that assemblages AI and AII of G. duodenalis provide the greatest potential zoonotic risk to humans. Horses may therefore constitute a potential source for human infection of Giardia either directly or via watersheds.     

Influence of climate and abundance on migration timing of adult Atlantic salmon (Salmo salar) among rivers in Newfoundland and Labrador

Evidence mounts for the influence of climate variability on temporal trends in the phenology of many organisms including various species of fish. Accordingly, we examined variation in adult Atlantic salmon Salmo salar run timing in thirteen Newfoundland and Labrador rivers where returns were monitored at fishways or fish‐counting fences. Run timing varied significantly among rivers with the median date of return differing by up to 5 weeks. Duration of runs was generally short with most adults returning over a period of three to 5 weeks. A mixed model analysis incorporating a first‐order autoregressive error structure was used to generalise changes in run timing among all monitored rivers. Results indicated that the median date of return has advanced by almost 12 days over a 35‐year interval from 1978 to 2012, while several individual rivers have advanced by almost 21 days. The influence of climate on median timing was evident when the simultaneous effects of both climate and salmon abundance were controlled. We found earlier runs associated with overall warmer climate conditions on the Newfoundland and Labrador Shelf. Results contrast with those from the north‐east Atlantic where Atlantic salmon are returning later in some rivers coincident with warming climate conditions.     

Modified plant architecture to enhance crop disease control: genetic control and possible value of upright fruit position in cucumber

Plant architectural features can facilitate disease control by creating unfavourable environments for pathogen growth or limiting pathogen contact with the host. In the case of Phytophthora capsici infection of cucumber (Cucumis sativus), the susceptible fruit typically lie in contact with soil under warm, moist conditions of a full canopy, an ideal location for exposure to inoculum and disease development. We have shown that increased row spacing, trellising and architectural variants that allow for more open canopies, such as shorter vines, reduced branching, or smaller leaves, can modify microclimate as assessed by temperature at the location of developing fruits. However, only trellising reduced infection rates (<3 % infection vs. 22 % for control). A cucumber accession PI308916, with extremely short internodes and resulting upright position of young fruit, also exhibited reduced disease. Fruit of PI308916 were susceptible when inoculated with P. capsici, indicating that reduced infection likely resulted from decreased soil contact. The compact trait is inherited as a single recessive gene, cp. Like many architectural traits, cp may be pleiotropic; PI308916-derived breeding lines were reported to have poor seedling emergence. PI308916 exhibited inconsistent apical hook formation that co-segregated with short internodes in F₂ and backcross generations, and disorganized internodal cortical cell arrangement. The combined phenotypes suggest either pleiotropy or very tight linkage of genes affecting hormone balance or cell division. The narrow chromosome region recently defined by QTL analysis includes candidate gene homologs of meristem related genes modulating cell division or spacing of lateral shoot organs and genes associated with brassinosteroid signaling or cytokinin content.     

Spawning success and early life history of longnose suckers in Great Lakes tributaries

Fish eggs and larvae are often subject to very high mortality, and variation in early life survival can be important for population dynamics. Although longnose suckers (Catostomus catostomus) are widespread in northern North America, little is known about their early life history. We examined fecundity and early larval survivorship during sucker spawning events in three small Lake Michigan tributaries. Although egg deposition varied 25% among spawning events, estimated larval export to the lake varied over 25,000‐fold from around 1000 to 26 million. Based on variation in environmental conditions across years, it appears that spring flow and temperature may be important determinants of egg survival to larval outmigration. Larval age data suggest that most individuals that survived to outmigration hatched during a 2‐day period despite adult spawning across at least 10 days. Most larvae spent <2 weeks in the stream and emigrated around the time of transition from endogenous to exogenous feeding before substantial growth occurred. In two of three cases, larvae drifted exclusively at night; however, high drift rates occurred during both day and night in the case where larvae were very abundant, suggesting density‐dependent drift behaviour. Our results indicate that survival in tributary streams from egg deposition to larval export is highly variable in longnose suckers. These large differences in early life survival may translate into variability in recruitment, thereby influencing population structure and dynamics.     

Mutations in U4atac snRNA, a component of the minor spliceosome, in the developmental disorder MOPD I

Small nuclear RNAs (snRNAs) are essential factors in messenger RNA splicing. By means of homozygosity mapping and deep sequencing, we show that a gene encoding U4atac snRNA, a component of the minor U12-dependent spliceosome, is mutated in individuals with microcephalic osteodysplastic primordial dwarfism type I (MOPD I), a severe developmental disorder characterized by extreme intrauterine growth retardation and multiple organ abnormalities. Functional assays showed that mutations (30G>A, 51G>A, 55G>A, and 111G>A) associated with MOPD I cause defective U12-dependent splicing. Endogenous U12-dependent but not U2-dependent introns were found to be poorly spliced in MOPD I patient fibroblast cells. The introduction of wild-type U4atac snRNA into MOPD I cells enhanced U12-dependent splicing. These results illustrate the critical role of minor intron splicing in human development.     

Simultaneous analysis of free phytosterols/phytostanols and intact phytosteryl/phytostanyl fatty acid and phenolic acid esters in cereals

An approach based on solid-phase extraction for the effective separation of free phytosterols/phytostanols and phytosteryl/phytostanyl fatty acid and phenolic acid esters from cereal lipids was developed. The ester conjugates were analyzed in their intact form by means of capillary gas chromatography. Besides free sterols and stanols, up to 33 different fatty acid and phenolic acid esters were identified in four different cereal grains via gas chromatography-mass spectrometry. The majority (52-57%) of the sterols and stanols were present as fatty acid esters. The highest levels of all three sterol and stanol classes based on dry matter of ground kernels were determined in corn, whereas the oil extract of rye was 1.7 and 1.6 times richer in fatty acid esters and free sterols/stanols than the corn oil. The results showed that there are considerable differences in the sterols/stanols and their ester profiles and contents obtained from corn compared to rye, wheat, and spelt. The proposed method is useful for the quantification of a wide range of free phytosterols/phytostanols and intact phytosteryl/phytostanyl esters to characterize different types of grain.     

Development of a whole-body cortisol extraction procedure for determination of stress in golden shiners, <Emphasis Type="Italic">Notemigonus crysoleucas</Emphasis>

Baitfish such as golden shiners are subjected to stress during harvesting, grading, and transport. Their small size makes it difficult to measure the stress response with the biological indicator cortisol using conventional assay methods for plasma. This paper examines the development and validation of methods for whole-body cortisol extraction from individual baitfish. Three types of extracts were tested: (1) an ethyl ether unaltered extract (UA); (2) an extract reconstituted in phosphate buffered saline (PBS); (3) an extract that had been increased in volume by the addition of food-grade vegetable oil (VO). These extracts were evaluated using validation tests with radioimmunoassays (RIA) and enzyme-linked immunosorbent assays (ELISA). The UA extract produced inadequate volumes of extract for multiple assays and could not be used for the determination of cortisol in a single fish. The PBS reconstitution method failed the precision recovery of serial dilutions (62.3%), linearity (R ²: 0.7864), and parallelism validation tests. The VO volume-boosting method passed all validation tests [intra-assay coefficent of variation (%CV): 16.3 for ELISA and 5.9 for RIA; inter-assay %CV: 10.3; spiked recovery: 102.0%; dilution recovery: 93.0%; linearity R ²: 0.9435; log of serial dilutions was parallel] and provided enough extract for multiple assays from an individual baitfish. Based on these results, we conclude that the VO volume-boosting method presents a means for determining cortisol from individual baitfish using either RIA or ELISA assays.