Evaluation of Sperm-Activating Solutions in Atlantic Salmon Salmo salar Fertilization Tests

The use of saline solutions was tested to determine their efficacy as replacements for ovarian fluid as sperm activators and to eliminate variability encountered with the use of Ovarian fluid. We tested fertilization rate of semen from eight males on Atlantic salmon Salmo salar eggs after five sperm-activating solutions and a non-activating saline were substituted for ovarian fluid. We used solutions shown acceptable for use with other salmonid species. The six solutions tested were a non sperm-activating phosphate-buffered saline (PBS, 7.2 g/L NaCl, 1.48 g/L Na₂HPO₄, 0.43 g/L K H₂PO₄), a Tris buffer (6.99 g/L NaCl, 3.63 g/L Tris and 2.42 g/L glycine), a Borax buffer (12.2 g boric acid/L in solution 1, 76 g disodium tetraborate/L combined 100:118, then 1 L combined with 3.7 L water and 18 g NaCl), and three solutions of 9.25 g/L (125 mM) NaCl buffered to pH 6.0, 7.5, and 8.9. The latter five solutions activated sperm immediately on contact, while PBS required additional water to activate sperm. The PBS solution was the least effective (mean percent eyed eggs 37.6%) for egg fertilization. The mean percent eyed eggs for the other five saline solutions (range 78% to 86%) were not significantly different. Sperm from one male provided significantly lower egg fertilization (39.6%) when compared with the other seven males (67.2–87.4% egg fertilization). Percent egg fertilization was not related to number of live sperm cells per egg. Our results show that osmotically-balanced sperm-activation solutions, even those with a pH range from 6.0 to 8.9 provide adequate media for fertilization of Atlantic salmon eggs. Fertilization in a deactivation saline and water reactivation of sperm resulted in low egg fertilization.     

QTL analysis of resistance to powdery mildew in hop (<Emphasis Type="Italic">Humulus lupulus</Emphasis> L.)

Hop with powdery mildew [HPM: caused by Podosphaera macularis (Wallr.) U. Braun & S. Takam.] results in significant losses in hop production by reducing yield and quality. One means of increasing yield and quality is the production of resistant hop lines. Breeding for resistance can be significantly improved and accelerated by use of marker-assisted selection. The objective of this preliminary study was to identify QTLs and markers for genetic resistance to HPM. A bi-parental mapping population between the resistant line “Newport” and susceptible line ‘21110M’. Phenotypic data was scored under controlled greenhouse conditions. Significant differences among offspring were observed and disease resistance did not follow a distinct binomial distribution, suggesting quantitative genetic control. Genotyping-by-sequencing resulted in approximately 375 K SNP markers, which were filtered down to 2263 markers mapped to 10 linkage groups. Interval Mapping identified four QTLs with one on linkage group 1 and three located on linkage group 6. Composite interval mapping identified three QTLs, all located on linkage group 6. Mixed linear models identified 15 markers associated with expression of resistance to HPM. Three of these 15 SNPs were also identified in QTL-CIM analysis. Evaluation of the scaffolds containing the significant SNP markers identified seven putative genes—several of which appear involved in disease resistance in other plant species. The SNP markers identified in this study still require validation in unrelated populations prior to implementation in breeding programs.     

Phosphorus utilization in rainbow trout (Oncorhynchus mykiss) fed practical diets and its consequences on effluent phosphorus levels

Excessive dietary phosphorous (P) concentrations in effluents from aquaculture present a major environmental problem. We therefore studied the effect of dietary P and vitamin D₃ on P utilization by rainbow trout-fed practical diets and on P concentrations in the soluble, particulate and settleable components of the effluent from fish tanks. Rainbow trout (average weight: 78 g, initial biomass: 13 kg in 0.7 m³ tanks) were fed for 11 weeks, practical diets that varied in total P, available P, and vitamin D₃ concentrations. Soluble, particulate (10–200 μm) and settleable (>200 μm) P in the effluent were sampled every 0.5–6 h for 1–3 days in the third and eleventh weeks of the experiment. Trout in all diets more than doubled their weight after 11 weeks. Increasing the concentrations of available dietary P from 0.24% to 0.88% modestly enhanced growth rate. Feed conversion ratio (FCR) and biomass gain per gram P consumed decreased as dietary P concentrations increased. Carcass P, daily P gain, and plasma P concentrations were lower in fish fed with low P diets. Soluble P concentrations in the effluent peaked immediately after and again 4–6 h after feeding, and is a linear function of available dietary P. No soluble P would be produced during consumption of diets containing less than 0.22±0.02% available P. Above this dietary concentration, soluble P would be excreted at 6.9±0.4 mg/day/kg for each 0.1% increase in available dietary P. Particulate P concentrations in the effluent were independent of dietary P concentrations. Settleable, presumably fecal, P concentrations tended to increase with dietary P concentrations. In trout fed with low P (0.24% available P, 0.6% total P) diets, 60% of total dietary P were retained by the fish and the remaining 40% were excreted in the effluent as settleable P (20–30%) and particulate or soluble P (10–20%). In trout fed with high P (0.59–0.88% available P; 0.9–1.2% total P) diets, 30–55% of total dietary P was retained by fish, and the remaining 15–25% appeared in the effluent as settleable P, 20–55% as soluble P, and 5–10% as particulate P. Vitamin D₃ did not affect fish growth nor effluent P levels. Physicochemical management of aquaculture effluents should consider the effect of diets on partitioning of effluent P, the peaks of soluble P concentration following feeding, and the contributions of particulate P to total P in the effluent. Increasing our understanding of how dietary P is utilized and is subsequently partitioned in the effluent can contribute significantly towards alleviating this important environmental and industry problem.     

Leaf litter decomposition in a chaparral ecosystem, Southern California

Decomposition losses from leaves of three evergreen chaparral species, scrub oak (Quercus dumosa), ceanothus (Ceanothus crassifolius), and manzanita (Arctostaphylos glauca), were quantified over a 2-y field exposure using litterbags. Changes in ash-free dry mass, C, and N were monitored at 2- to 6-month intervals at four replicate sites composed of patches of these three chaparral species. Three proximate C fractions were extracted from fresh and decomposing litter samples: polar and non-polar extractives (EXT), acid-solubles (ACID), and acid-insolubles (KLIG). The chemical structure of fresh and decomposed litter was additionally characterized using high-resolution solid-state ¹³C NMR spectroscopy, while morphological properties were examined by scanning electron microscopy (SEM). After 2 y, the litters had lost between 20.7%±1.2 (Ceanothus) and 35.2%±6.8 (Quercus) of their original ash-free dry mass. The manzanita decomposed at a significantly faster rate than the other two litter types during the first few months of field exposure. Yet, after 2 y, mass loss was greater for the oak. Differences in decomposition rates could not be accounted for based on a single litter quality index. Fresh manzanita exhibited a significantly higher N content, which could explain its initially faster decay rate. Fresh oak litter, on the other hand, had a relatively high ACID and O-alkyl C (O-ALK) content, which may have been responsible for its decay pattern. Fresh ceanothus contained a relatively low KLIG content, yet it decomposed more slowly than the two other species. The solid-state ¹³C NMR spectra of the ceanothus litter had two peaks characteristic of proanthocyanidins, which likely contributed to the recalcitrance of this litter type. SEM revealed that ceanothus leaf surfaces were left nearly unchanged after field exposure. In comparison, the oak and manzanita leaf surfaces were pitted and covered by microbial growth to the point of being unrecognizable. Taken together, our results indicate that a combination of biological, physical and chemical factors need to be examined to clarify the different decomposition rates and patterns of these three chaparral species.     

Population dynamics of Enchytraeidae (Oligochaeta) in different agricultural systems

Organic matter and abiotic conditions seem to influence distribution patterns of Enchytraeidae. In this study effects of changes in management practices on population dynamics of enchytraeids were determined. At two sites (in Athens and Griffin, GA) parts of a fescue (FE) field were converted into conventional tillage (CT) and no-tillage (NT) plots and changes in densities and depth distribution of enchytraeids were followed for 3 years. A site at Watkinsville, containing various soil textures and characterised by very low organic carbon content, which was converted into no-tillage 4 years earlier, was also sampled. Significant reductions in enchytraeid densities, after conversion of fescue into CT, were only found at Griffin. The management practices affected the vertical distribution of enchytraeids. In fescue and NT more enchytraeids were found in the 0–5cm than in the 5–15cm layer. In conventional tillage fields enchytraeids were more evenly distributed over the profile or more abundant in the 5–15cm layer. Management also affected the timing of population dynamics in the different plots. At two sample dates high abundances were found in CT plots only, not in any of the other plots. Enchytraeids were larger at Athens than at Griffin and Watkinsville and contained more soil particles in their gut. At Athens enchytraeids will presumably contribute more to the development of soil structure than at the other sites. We conclude that management affects vertical enchytraeid distributions in soil and changes the timing of population dynamics. Received: 7 January 1996     

Virus maturation involving large subunit rotations and local refolding

Large-scale conformational changes transform viral precursors into infectious virions. The structure of bacteriophage HK97 capsid, Head-II, was recently solved by crystallography, revealing a catenated cross-linked topology. We have visualized its precursor, Prohead-II, by cryoelectron microscopy and modeled the conformational change by appropriately adapting Head-II. Rigid-body rotations ( approximately 40 degrees) cause switching to an entirely different set of interactions; in addition, two motifs undergo refolding. These changes stabilize the capsid by increasing the surface area buried at interfaces and bringing the cross-link-forming residues, initially approximately 40 angstroms apart, close together. The inner surface of Prohead-II is negatively charged, suggesting that the transition is triggered electrostatically by DNA packaging.     

Porphyrins are not present in feline ocular tissues or corneal sequestra

Objective To determine if feline lacrimal glands, glands of the third eyelid, corneas, and corneal sequestra contain porphyrins, which could be responsible for the brown/amber discoloration of corneal sequestra and tears in affected cats. Procedures Samples of grossly normal cornea, lacrimal gland, gland of the third eyelid, and sequestra obtained via keratectomy were collected. Porphyrin concentrations of the homogenate were determined by spectrofluorometry with protoporphyrin IX and coproporphyrin III dihydrochloride used as standards. A hamster harderian gland was used as a positive control. Results Normal tissues were harvested from one eye each of 14 nonclient owned, adult, mixed‐breed, short‐hair cats euthanized for reasons not associated with this study. Eighteen sequestra were acquired from cats undergoing unilateral lamellar keratectomies. Breeds of the affected cats included eight Himalayan, five domestic shorthair, and one each of four other breeds. Only the positive control and standards contained levels of porphyrins above background. All feline samples examined were histologically normal with no evidence of porphyrins. Conclusions Porphyrins are absent in normal feline lacrimal glands, corneas, and corneal sequestra. Porphyrins do not appear to be the cause of the brown/amber color of feline corneal sequestra.