Application of sperm cryopreservation in selective breeding of the Pacific oyster,Crassostrea gigas (Thunberg)

The robustness of Pacific oyster, Crassostrea gigas (Thunberg), sperm cryopreservation in the context of selective breeding based on family lines was investigated. Irrespective of egg density, high fertilization success was achieved with cryopreserved sperm when sperm:egg ratios of 1000:1 to 10 000:1 were used. Variation among replicate runs on the same oyster batches was minimal, indicating that cryopreservation and larval rearing procedures were repeatable. Twenty independent single male–female crosses were made to assess the utility of cryopreserved sperm in selective breeding. The fertility of unfrozen sperm was generally a poor predictor of cryopreserved sperm fertility. Based on D‐larval yields, 17 of the 20 crosses were likely to yield adequate spat for selective breeding (>10⁵ D‐larvae from 1 million eggs), two were marginal (5 × 10⁴ D‐larvae) and one was inadequate (4 × 10³ D‐larvae). An alternative fertilization strategy to improve D‐yield from a given number of sperm was then tested. Fertilizing 10 million eggs at a sperm:egg ratio of 200:1 increased the total D‐yield when compared with fertilizing 1 million eggs at a sperm:egg ratio of 2000:1 for the same male–female pair. We conclude that, despite wide variation in fertility, cryopreserved sperm is useful for family production.     

Early-successional vegetation changes after roadside prairie restoration modify processes related with soil functioning by changing microbial functional diversity

Because of their rapidly changing vegetation dynamics and harsh environmental conditions, roadside prairies in semi-arid regions represent an exceptional study system in which to investigate the effects of plant-soil interactions on ecosystem functioning. We conducted a two-year field experiment on two roadside embankments in semi-arid central Spain differing in construction age to answer the following questions: (i) do commonly used restoration treatments (hydroseeding, fertilization and irrigation) affect soil microbial functional diversity and processes related to soil functioning (basal respiration, total N and P and in situ N availability rate)? (ii) what portion of plant effects on processes related to soil functioning is mediated indirectly by microbial functional diversity? Except for a small and negative irrigation effect on the microbial functional diversity in the three-year old site, the restoration treatments employed did not affect this variable. Fertilization increased plant diversity, an effect likely mediated by the enhanced soil nutrient availability with this treatment at early stages of secondary succession. In contrast, hydroseeding did not affect processes related to soil functioning. The total effect of the plant community on these processes was higher than that of the microbial functional diversity alone, suggesting that the studied slopes are to the greater extent regulated by plants. However, soil microbes are a key proximate influence in the system, as the indirect effects of plant community on soil functioning processes mediated by soil microbes represented 37-41% of the total plant effects observed. Our results indicate that the restoration of recently built slopes can potentially be improved with treatments that promote plant compositional shifts, such as fertilization, or alter soil function, such as the enhancement of soil microbial functional diversity. They also highlight that plant-soil interactions are an important process that can be manipulated for restoration purposes in early-successional stages, especially in nutrient-poor semi-arid ecosystems.     

A recombinant vaccine expressing a mammalian Mycobacterium sp. antigen is immunostimulatory but not protective in striped bass

A recombinant vaccine was constructed for piscine mycobacteriosis utilizing a Brucella abortus strain RB51 vector expressing a mammalian Mycobacterium sp. 85A antigen. Juvenile striped bass were inoculated with the resulting construct at doses equivalent to 10(6), 10(7), 10(8), 10(9), and 10(10) colony-forming units/fish. Blood and tissue samples from these fish demonstrated significant specific humoral and cell-mediated immune responses towards the 85A antigen in a dose-dependent manner. However, survival studies determined that inoculated fish failed to demonstrate cross-protective responses after live Mycobacterium marinum challenge 70 days post-inoculation.     

Research needs for restoring seasonal tropical forests in Thailand: accelerated natural regeneration

Accelerated natural regeneration (ANR) is a relatively cheap method of reforestation, which encourages natural establishment of indigenous trees and shrubs. It requires a low input of labour, but a high input of ecological information. In this paper, the knowledge required to predict and manipulate the natural regeneration of seasonal tropical forest is reviewed and areas in need of further research are identified. Regeneration will be influenced by five groups of potentially limiting factors; site resources (soil and microclimate); competition with weeds; site disturbance; occurrence of established woody plants or their propagules; seed dispersal by wild animals and birds. This paper considers each of these, their interactions with seasons, and each other. Collation of existing information on these topics, combined with the suggested further research, should facilitate the creation of tools that will enable practitioners to judge the regeneration potential of sites and to select the most appropriate ANR techniques.     

Esterification and hydrolytic activities of Candida rugosa lipase isoform 1 (LIP1) immobilized on celite 545, duolite A7, and sephadex G-25

The esterification and hydrolytic activities of free and immobilized Candida rugosa lipase isoform 1 (LIP1) were investigated. Esterification activity was determined by reacting caprylic acid with glycerol in the presence of molecular sieves (30%, w/w), and the volume of 1.0 M NaOH consumed by the reaction products upon titration was used to calculate esterification activity. Caprylic acid was also reacted with cottonseed oil, and the amount of caprylic acid incorporated after 12 h of reaction was determined. Results indicated that LIP1 had little esterification activity, which was not significantly improved upon immobilization. Hydrolytic activity was determined by incubating tricaprylin emulsion (15%, w/w) with the respective lipases for 60 min, and the reaction products were titrated against 0.5 M NaOH. LIP1 showed hydrolytic activity comparable to Lipozyme RM IM. The hydrolytic activity improved significantly upon immobilization. Immobilization on Celite 545 produced the highest increase in hydrolytic activity.     

Carbonate mineral filtrants with new surfaces reduce alkalinity in seawater and artificial seawater: Preliminary findings

Silica sand (silica), coral (aragonite), and oyster shell (calcite) were ground to similar particle sizes and placed in seawater and artificial seawater (GP2 Medium). Alkalinity and pH values of the artificial seawater decreased substantially over 24 h when in contact with coral and oyster shell; the effects in seawater were minor. Once alkalinity has been reduced, the maintenance of stable pH at values typical of seawater is made more difficult. The data, which are preliminary, have practical application.     

The effect of introgressions of wheat D-genome chromosomes into 'Presto' triticale

Hexaploid triticale (X Triticosecale Wittmack) (2n= 6x= 42, AABBRR) and wheat (Triticum aestivum L.) (2n= 6x= 42, AABBDD) differ in their R and D-genomes. This produces differences in both agronomic and end-use quality characteristics. Our objective was to determine how introgressions of individual chromosomes from the D-genome of wheat affect these characteristics of a winter triticale 'Presto'. We studied the effects of 18 D-genome chromosome substitution lines, 15 sib-lines as controls, and five check cultivars at Lincoln, NE in 1996, using a randomized complete block design with two replications. The experiment was repeated at Lincoln and Mead, NE in 1997 and 1998 with 15 substitution lines that survived the first winter in Lincoln, along with their 12 control sibs and five check cultivars. Few D-genome chromosomes had positive effects. Chromosomes 2D, 4D, and 6D significantly reduced plant height when substituted for 2R, 4B, and 6R, respectively. No grain yield increases were associated with any of the D-genome chromosomes tested, but three substitutions decreased the grain yield. Depending on the allele of the hardness gene present, chromosome 5D increased or decreased kernel hardness when substituted for 5R or 5A, respectively. Introgressions of chromosomes 1D and 6D improved end-use quality characteristics of Presto. These results suggest that apart from beneficial effects of individual loci located on the D-genome chromosomes, no major benefit can be expected from D-genome chromosome substitutions.