Semen cryopreservation of salmonid fishes: influence of handling parameters on the postthaw fertilization rate

Using the cryopreservation method of Lahnsteiner, Berger, Weismann & Patzner (1995, Aquaculture Research 26 , 801-807) the influence of allowable variations of methodical parameters (storage of semen before cryopreservation, dilution ratios in the extender, equilibration in the extender, cooling rates, storage of deep-frozen semen in liquid nitrogen, storage of frozen/thawed semen, minimal sperm/egg ratio) was investigated under the aspect of routine utilization. Under optimized experimental conditions, fertilization rates were 90-100% of controls in Oncorhynchus mykiss (Walbaum), Salmo trutta L. f. lacustrisSalmo truttaL.f. fario and Salvelinus fontinalis (Mitchill). The following results were obtained: 1. Storage of untreated semen for more than 1 h before cryopreservation decreased the postthaw fertility. 2. Equilibration of semen up to 20 min in the extender did not affect the postthaw fertility. 3. Optimal dilution ratio of semen in the extender was threefold in Oncorhynchus mykiss and Salvelinus fontinalis. Lower dilution ratios decreased the postthaw fertility, higher dilution ratios did not affect the postthaw fertility. In Salmo trutta f. lacustris and Salmo trutta f. fario, which have a higher sperm density, optimal dilution ratio of semen in the extender was fivefold to sevenfold. 4. In Oncorhynchus mykiss, as in Salmo trutta f lacustris and Salmo trutta f. fario, the optimal freezing height was at 1.5 cm above the level of liquid nitrogen (-110 ± 2^oC); in Salvelinus fontinalis it was 2.5 cm above the level of liquid nitrogen (-92 ± 2^oC). Changes in the freezing height of 0.5 cm (about 10^oC) resulted in a significant decrease of postthaw fertility. 5. Storage of deep-frozen semen for up to 370 days in liquid nitrogen had no influence on its postthaw fertilization rate. 6. Storage of frozen/thawed semen for 30 s before insemination significantly decreased its postthaw fertility. 7. Reliable minimal sperm:egg ratio to obtain fertilization rates of 90-100% of control was 3-5 X 10⁵ spermatozoa egg^-1.     

Barriers and flow as limiting factors in the spread of an invasive crayfish (Procambarus clarkii) in southern California streams

Invasive crayfish are a major threat to stream ecosystems, yet research has seldom identified successful ways of preventing their spread. Thirty-two stream sections were surveyed during 2000 and 2001 in the Santa Monica Mountains of southern California to determine the distribution of the invasive crayfish Procambarus clarkii. Streams with large barriers (waterfalls, culverts) often did not have crayfish present upstream of barriers. A mark-recapture study indicated that P. clarkii moved both up and downstream between pools, but that barriers significantly reduced movement between pools. Seasonal high flow velocities likely increase passive movement downstream and reduce movement upstream. Results indicate that crayfish mainly spread downstream from a point of colonization and are restricted in their movement to adjacent upstream sections by both natural and artificial barriers. We suggest management strategies for removing invasive crayfish and reducing their spread by focusing on smaller stream segments that are bounded by a downstream barrier and by timing removal efforts to follow large flow events.     

Nutrient release from decomposing leaf litter of temperate deciduous forest trees along a gradient of increasing tree species diversity

In the litter of six deciduous tree species (Fagus sylvatica, Tilia spp., Fraxinus excelsior, Carpinus betulus, Acer pseudoplatanus and Acer platanoides) and in stand-specific litter mixtures, we compared mass loss and nutrient release across diversity levels along a gradient of decreasing proportion of Fagus in stands with similar environmental and physical soil conditions. The litterbag studies ran over 22 months. The decomposition rate constants (k) of the temperate forest species ranged from k = 0.5 for Fagus to k = 1.5-2 for all other tree species. In Fagus, k was closely negatively correlated with the thickness of the litter layer and positively correlated with soil pH and isopod abundance. k was significantly higher in the mixed species stands (except for Carpinus and Fraxinus) and was positively correlated with earthworm abundance. Over the whole incubation time, nutrient amount and release rates of N, P, K, Ca and Mg as well as C-related ratios showed significant differences between tree species but no consistent differences among the diversity levels. Initial C-related nutrient ratios of the leaf litter and abundance of mesofauna and macrofauna were correlated with the length of time lag before nutrient release. We conclude that the mere number of tree species is not the main driver of nutrient release and decomposition processes, but that key groups of the decomposer fauna as well as the characteristic traits of the individual tree species are decisive.     

Aromatic cross-links in insect cuticle: detection by solid-state 13C and 15N NMR

Cross-polarization magic-angle-spinning nuclear magnetic resonance spectroscopy has been used to determine insect cuticle composition and cross-link structure during sclerotization or tanning. Unsclerotized cuticle from newly ecdysed pupae of the tobacco hornworm, Manduca sexta L., had a high protein content with lesser amounts of lipid and chitin. Concentrations of chitin, protein, and catechol increased substantially as dehydration and sclerotization progressed. Analysis of intact cuticle specifically labeled with carbon-13 and nitrogen-15 revealed direct covalent linkages between ring nitrogens of protein histidyl residues and ring carbons derived from the catecholamine dopamine. This carbon-nitrogen adduct was present in chitin isolated from cuticle by alkaline extraction and is probably bound covalently to chitin. These data support the hypothesis that the stiffening of insect cuticle during sclerotization results primarily from the deposition of protein and chitin polymers and their crosslinking by quinonoid derivatives of catecholamines.     

A comparative molecular field analysis study of obtusifoliol 14α-methyl demethylase inhibitors

This report describes the development of a Comparative Molecular Field Analysis (CoMFA) model from a set of obtusifoliol 14α-methyl demethylase (DM) inhibitors to aid in the design of herbicides targeting sterol biosynthesis. CoMFA is a three-dimensional (3-D) quantitative structure–activity relationship (QSAR) method that is useful in the probing of receptor binding sites when experimental structure data are unavailable. Conformational analysis and SAR of some rigid and active analogs were used to build the initial model using the active analog hypothesis. The model was subsequently used to design compounds that retain the active site shape requirements, but incorporate physical properties that favor soil-applied herbicidal action. In addition, a second-generation CoMFA model incorporating the newly designed inhibitors was developed and represents the current understanding of the DM binding site. This model was derived from a pharmacophore developed from two methods, the active analog approach as well as from the Catalyst program. The fact that two independent methods produced a similar pharmacophore strengthens the validity of the model. © 1999 Society of Chemical Industry     

Gross Morphometry of the Bovine Placentome during Gestation

Forty-seven pregnant uteri were collected from an abattoir at eight chronological stages of gestation. Assessments were made of placentome number, weight, length shape and ratio of foetal to maternal tissue by weight. The study showed that there was a great variation in number, size, weight and shape of placentomes. There was no significant change in the total number of placentomes during gestation. Both mean weight and length increased significantly during gestation, but there was a significant change in the relationship between the two as gestation progressed. There was no correlation between the number of placentomes and the mean weight or length of the placentomes. The development of the placenta in the non-pregnant horn was significantly different from that of the pregnant horn, with fewer, smaller, lighter placentomes. However, there was no evidence that placentome development in the non-pregnant horn was affected by placentome development in the pregnant horn. The proportion of the placentome that was maternal tissue increased during gestation, but there was no significant effect of placentome number, or individual placentome weight or length on this proportion. Two placentome shapes were identified, but no effect on foeto-maternal proportion or relationship between shape and total placentome number or weight was identified.     

Food choice and impact of food sources from farms on blood coagulation in rodenticide resistant Norway rats

Rodents are mostly controlled by using anticoagulant rodenticides to minimise adverse effects along the food production chain. Genetic resistance to anticoagulant compounds occurs in populations of Norway rats (Rattus norvergicus) and other rodents throughout the world. This can be problematic for control operations and put predators and scavengers at risk because resistant prey individuals carry high anticoagulant loads. Single nucleotide polymorphisms (SNPs) in the vitamin K reductase complex subunit 1 (VKORC1) gene that are associated with rodenticide resistance are related to several pleiotropic effects including increased vitamin K requirement. The uptake of dietary vitamin K available from food sources present on farms including corn silage and garden herbs may mitigate such effects. We tested the preference for food rich in vitamin K1/3 in Norway rats susceptible or resistant to bromadiolone due to the homozygous amino acid variant Tyr139Cys. We also studied the effect of vitamin K consumption from several food sources on blood clotting in bromadiolone resistant and susceptible Norway rats. There was no general preference of rats for food sources rich in vitamin K. Surprisingly, there was no effect of vitamin K uptake on blood clotting times. However, the consumption of 4 of 5 corn silages prevented the rise in blood clotting times that otherwise occurred in vitamin K deficient bromadiolone resistant rats. This seemed to be due to substances with vitamin K activity present in silage and may contribute to the sustaining and spreading SNPs that are related to rodenticide resistance. As corn silage is becoming increasingly available to rats due to the expansion in generating bio-energy from corn silage, there may be a future increase in the occurrence of anticoagulant resistant Norway rats and associated problems in agro-ecosystems.     

Technological Advances to Address Current Issues in Entomology: 2020 Student Debates

The 2020 Student Debates of the Entomological Society of America (ESA) were live-streamed during the Virtual Annual Meeting to debate current, prominent entomological issues of interest to members. The Student Debates Subcommittee of the National ESA Student Affairs Committee coordinated the student efforts throughout the year and hosted the live event. This year, four unbiased introductory speakers provided background for each debate topic while four multi-university teams were each assigned a debate topic under the theme ‘Technological Advances to Address Current Issues in Entomology’. The two debate topics selected were as follows: 1) What is the best taxonomic approach to identify and classify insects? and 2) What is the best current technology to address the locust swarms worldwide? Unbiased introduction speakers and debate teams began preparing approximately six months before the live event. During the live event, teams shared their critical thinking and practiced communication skills by defending their positions on either taxonomical identification and classification of insects or managing the damaging outbreaks of locusts in crops.