Effects of Serum-Free In Vitro Maturation of Bovine Oocytes on Subsequent Embryo Development and Cell Allocation in Two Developmental Stages of Day 7 Blastocysts

Maturation of oocytes and the subsequent outcome of the in vitro production (IVP) are affected by the composition of in vitro maturation (IVM) medium. To determine the use of serum interfering with effects of single molecules, we aimed at developing simplified IVM medium. The experimental IVM media were: (1) M199-medium supplemented with hormones and serum (control), (2) as 1 but serum was substituted with fatty acid-free serum albumin (FAFBSA) and (3) M199-medium without hormonal and serum supplementation (M199). The quality of embryos was assessed on day 7 by morphology and cryotolerance, as well as by Terminal deoxynucleotidyl Transferase Biotin-dUTP Nick End Labeling (TUNEL) and differential staining. Results showed that the nuclear maturation was suppressed in M199 group alone. Embryo cleavage and development rates, and the proportion of quality 1 blastocysts were lower in the FAFBSA and M199 groups compared to the control. Differences in the cell allocation of fresh embryos were observed at the blastocyst stage, but not at the expanded blastocyst stage. The control group blastocysts had larger number of cells allocated to the inner cell mass (ICM), and the FAFBSA group blastocysts larger apoptotic cell proportion compared to the blastocysts derived from other groups. After cryopreservation, the reduction of ICM proportion and increase of apoptotic cell proportion of embryos were equal between the experimental groups. In conclusion, exclusion of serum from the IVM media reduces embryo development and may cause perturbations in blastocyst development. Differences in the cell allocation of blastocysts between IVM media may appear only when the developmental stages are taken into account.     

Aggressiveness of Phytophthora infestans on detached potato leaflets in four Nordic countries

Potato fields in Denmark, Finland, Norway and Sweden were sampled for single-lesion isolates of Phytophthora infestans . The aggressiveness of the isolates was determined on detached leaflets of potato cvs Bintje (susceptible) and Matilda (moderately resistant). The aggressiveness tests were carried out in the respective home countries of the isolates, with the exception of the Danish isolates. Fifteen Danish isolates were studied in each of the other three countries, including five isolates tested in all three laboratories. Results obtained from the Danish isolates revealed substantial differences between the test laboratories for infection efficiency, lesion growth rate and sporulation capacity on detached leaflets. When the laboratory effect was taken into account, the differences in aggressiveness between the countries were generally small or inconsistent between the test cultivars and epidemiologically insignificant. By contrast, variation among isolates within countries was substantial. The magnitude of the variation depended on country and cultivar. Maximal variation for the means of the isolates was between 89 and 185 h for latent period, between 100 and 1297 sporangia mm⁻² for sporulation capacity and between nearly zero and 6 mm day⁻¹ for lesion growth rate. Typically less than 1% of sporangia were able to cause infections, except in Norway. These extraordinarily low values may be an artefact of the testing method. High variation in results between the test laboratories emphasizes the need for caution when comparing results obtained by different research groups.     

Population and behavioural level responses of arable soil earthworms to boardmill sludge application

The response of earthworms to soil application of boardmill waste sludge was quantified in field and laboratory experiments. The influence of one application of 6×10⁴ tonnes ha⁻¹ of unamended sludge was tested against no application on silty-clay arable soil. After 2 years, results in stubble-cultivated soil showed a 1.7 times lower density of earthworms where sludge was added, whereas there was no difference in average earthworm fresh weight. In direct-drilled soil, there was an indication of lower average numbers of Lumbricus terrestris middens when sludge was applied. Negative impacts in the field may have been due to indirect negative effects of the sludge application. In the laboratory, habitat choice tests were undertaken with two common species from the field site using sludge-based mulch and fertiliser products. Aporrectodea caliginosa showed no discernible preference between soil and soil mixed with unamended sludge (mulch), but favoured soil over soil mixed with sludge and chicken manure (fertiliser). Tests with L. terrestris showed a similar pattern. The preference of L. terrestris for the two products was compared with that for chopped barley straw by direct observation of foraging behaviour. Differences in foraging time of L. terrestris for different feeds were not significant, but mass of straw collected was significantly greater compared with either type of sludge treatment.     

Decreased nitrogen rates and irrigation effect on celery yield and internal quality

The effect of decreased nitrogen rates (90, 60 and 30 kg N/ha) with and without irrigation on celery yield quantity and internal quality were studied in field experiments in 1993 and 1994. The decreased nitrogen rates reduced yields in both years. In the dry and warm conditions of 1994 nitrogen × irrigation interaction was clearly observed; irrigation increased yield more at higher nitrogen rates than at lower nitrogen rates. In the rainy conditions of 1993 irrigation did not effect the yield level.On the other hand, in both years, decreased nitrogen rates increased dry matter, vitamin C and dietary fiber contents, and nitrogen had a minor effect on total sugar content. In 1994 alpha- and beta carotene and thiamin were also analyzed but nitrogen had no effect on them. Irrigation increased dry matter and total sugar contents at the two lowest nitrogen rates in 1994, but decreased vitamin C contents in both years. There was also slight evidence that irrigation might decrease alpha- and beta-carotene and thiamin contents.     

The Relation between Stand Density and the Structure of Spring Rape (Brassica Napus L.)

The relation between stand density and structure of spring rape ( Brassica napus L.) was described with the aid of comprehensive measurements of structure. The structural components measured were plant height, stem diameter at root collar, site of the lowest pod on the main raceme, number of pods on the main raceme, site of the lowest branch on the main stem, number of primary branches and number of pods on the branches. The experiment was conducted at two nitrogen levels, 110 and 180 kg ha', and was replicated in 3 years, 1988, 1989 and 1990. Stand density was varied by using five different seeding rates and the number of plants m^-2 evaluated immediately prior to harvest was used to express the stand density. In the years considered, the densities varied between 16 and 520 plants m^-2. The relationship between plant density and rape plant structure was adequately described each year by a mukivariate second degree polynomial model. Varying the nitrogen application rate seemed to have no influence on this relationship. Denser plant stands produced thinner and shorter plants, the shortening occurring in the pod-producing section of the main raceme. Further, with increasing stand density, the number of branches decreased as did the number of pods on the branches and on the main raceme. These changes in plant structure retarded at densities over 150—200 plants m^-2. This seems to be the minimum density that should be achieved in rape stands.     

Screening for fungal antagonists of seed-borne Fusarium culmorum on wheat using in vivo tests

A screening system is described for identifying fungal antagonists from soil which are effective in controlling seed-borne Fusarium culmorum on wheat (T. aestivum L.). The procedure comprised three consecutive in vivo tests done under controlled conditions using three different plant growing media. Altogether 1701 fungal isolates from Finnish field soils were screened for biocontrol activity. 210 fungal isolates that had advanced to various stages in the screening system were also included in a field experiment. The isolates that were the most effective in controlling seedling blight under natural conditions belonged to the genus Gliocladium. The screening tests separated Gliocladium spp. rather clearly from the rest of test fungi and thus the overall agreement among the results was good. The assays run under controlled conditions did not efficiently predict which of the Gliocladium isolates would be best in protecting wheat seedlings under field conditions.