Production of Lambs After the Transfer of Fresh and Cryopreserved in vitro Produced Embryos from Prepubertal Lamb Oocytes and Unsorted and Sex-sorted Frozen-thawed Spermatozoa

Cumulus-oocyte complexes from hormone-stimulated 3-4-week-old (n=43) and 6-7-week-old (n=12) prepubertal lambs were matured in vitro and incubated with unsorted, or X- or Y-spermatozoa separated with a high-speed cell sorter (SX MoFlo)frozen-thawed. Presumptive zygotes were then cultured to the blastocyst stage, and transferred to recipients fresh or after cryopreservation (frozen). Oocyte cleavage was higher (p <0.05) with unsorted (515/926, 55.6%) than X- or Y-spermatozoa (261/672, 38.8% and 229/651, 35.2%, respectively) and blastocyst formation (% zygotes) by Day 9 of in vitro culture was lower (p <0.05) for X- (102/261, 39.1%) than unsorted spermatozoa (249/515, 48.3%), but did not differ between Y-spermatozoa (103/229, 45.0%) and unsorted spermatozoa, or between X- and Y-spermatozoa (p >0.05). For fresh embryos, survival to term was 50.0% (3/6) for unsorted, 0.0% (0/6) for X- and 16.7% (1/6) for Y-spermatozoa-derived embryos (p >0.05), and for frozen embryos was 4.0% (2/50) for unsorted, 9.1% (2/22) for X- and 2.9% (1/34) Y-spermatozoa-derived embryos (p >0.05). Of the two lambs born from X-spermatozoa-derived embryos, one was female (50%), and from the two Y-spermatozoa-derived lambs, both were male (100%), demonstrating that lambs can be produced after the transfer of fresh and cryopreserved IVP embryos derived from prepubertal lamb oocytes and frozen-thawed sex-sorted sperm.     

Silage and total mixed ration hygienic quality on commercial farms: implications for animal production

Implications of silage hygienic quality for animal production were investigated on forty‐five dairy farms in South West England. Samples of grass and maize silages and of total mixed rations (TMR) were obtained together with information on silage technology, herd size and animal production. Samples were analysed for mycotoxins, bacteria, yeasts, moulds and chemical composition. Thirteen mycotoxins were assayed, but none were detected in the samples of grass silage. However, mycotoxins were found in 0·9 of all maize and other silage samples, with deoxynivalenol and zearalenone predominating. There was no relationship between total mycotoxin concentration and mean lactation milk yield per cow. Enterobacteria counts tended to be higher in maize silage than in grass silage and higher still in TMR – a cause for concern. There were no relationships between mould counts and mycotoxin concentrations in silages, implying that mycotoxins may have been produced in the field pre‐ensiling.     

Relationships between soil and foliar nutrients in young densely planted mini-plots of Pinus radiata and Cupressus lusitanica

The long-term nature of forest crop rotations makes it difficult to determine impacts of forestry on soil nutrients that might be depleted by forest growth. We used small scale, highly stocked plots to compress the length of the rotation and rapidly induce nutrient depletion. In the study, two species (Pinus radiata D. Don and Cupressus lusitanica Miller) are compared under two disturbance regimes (soil undisturbed and compacted), and two fertiliser treatments (nil and plus fertiliser), applied in factorial combination at 33 sites, covering the range of climatic and edaphic variation found in plantation forests across New Zealand. To assess our ability to rapidly highlight important soil properties, foliar nutrient concentrations were determined 20 months after planting. It was hypothesised that the densely planted plots, even at a young age, would create sufficient pressure on nutrient resources to allow development of relationships between properties used as indicies of soil nutrient availability and foliar nutrient concentrations. For both species significant relationships between foliar nutrients and 0–10 cm layer soil properties from unfertilised plots were evident for N (total and mineralisable N) and P (total, acid extractable, organic, Bray-2 and Olsen P). With the exception of Ca in C. lusitanica, foliar K, Ca and Mg were correlated with their respective soil exchangeable cation measures. The results thus confirm the utility of the experimental approach and the relevance of the measured soil properties for forest productivity.

In unfertilised plots foliar N and P concentrations in P. radiata exceeded those in C. lusitanica, the differences being eliminated by fertiliser application. Foliar N/P ratios in P. radiata also exceeded those in C. lusitanica. In contrast to N and P, foliar K, Ca and Mg concentrations were all higher in C. lusitanica, the difference being particularly marked for Ca and Mg. P. radiata contained substantially higher concentrations of the metals Zn, Mn and Al than C. lusitanica, whereas the latter contained higher B concentrations. Possible reasons for differences between species in foliar nutrient concentrations are discussed.     

Volcanic ash and its clay mineralogy at Cape Hoskins,New Britain,Papua New Guinea

Volcanic ash falls, the oldest laid down approximately 2500 years ago, are described and correlated wherever possible.Most of these ash deposits are thought to have been derived from the Garbuna volcano, located about 40–50 km from Cape Hoskins.Clay mineral data show with increasing age a weathering sequence in the beds in which allophane changes to halloysite. Charcoal data indicate that large amounts of halloysite are present in fossil soil horizons dated between 300 and 2000 years B.P. This compares with a date of 8000–9000 years from Japan,whereas data from St. Vincent indicate that halloysite can be formed under humid tropical conditions within 4000 years.     

Development and evaluation of a rapid absorbed enzyme immunoassay test for the diagnosis of Johne''s disease in cattle

An absorbed enzyme immunoassay (EIA) test for Johne's disease in cattle was developed in which absorption of cross-reacting antibodies occurred as a rapid reaction in solution rather than overnight with whole organisms and a subsequent centrifugation step. Total test time was reduced to less than 2 h with a minimum of manipulations. The test was evaluated in cattle herds from Johne's disease-endemic and Johne's disease-free regions of Australia. Specificity was 99.8%. Calculations of sensitivity were affected by the history of the herd under test. However, the EIA detected in excess of 80% of animals before onset of clinical disease and 65% of faecal shedders were EIA positive on, or before, first detection of Mycobacterium paratuberculosis in their faeces. The test should aid epidemiological studies and be a useful tool in the management and control of Johne's disease.     

Some effects of diallyl bisulphide on sclerotia of Sclerotium cepivorum: Possible novel control method for white rot disease of onions

Sclerotium cepivorum (the causal agent of white rot of onions) persists in soil in the form of sclerotia whose germination is triggered by volatile sulphides and thiols released into the soil from roots of the genus Allium. The effects of treatment of the soil with diallyl disulphide (DADS) on the germination of these sclerotia has been studied, because if the germination can be triggered in the absence of the host crop, control of the fungus could be achieved. Five isolates of the fungus responded similarly but marked effects of DADS concentration and of the time-of-year of application were discovered, poor results being obtained with summer applications of DADS. Promising results were obtained at other times of the year with DADS flooded on to land containing sclerotia. The effectiveness of DADS treatment was shown to be related to the rate at which the material disappeared from soil, which was itself dependent upon soil temperature.     

Intraspecific olive diversity assessed with AFLP

Amplified fragment length polymorphism (AFLP) was used to study diversity within and among Spanish olive varieties. A high degree of diversity was found among the varieties present in each growing region. Olive oil production and quality relies on appropriate cultivar selection as well as good orchard management. Production based only on a few superior cultivars would result in improved yield, oil quality, and production management. Amplified fragment length polymorphism were evaluated as a tool to identify the intraspecific and intravarietal diversity of olive. Amplified fragment length polymorphism analysis of 38 accessions belonging to 10 cultivars using six primer combinations produced 106 polymorphic bands. Results were analyzed for similarity among accessions via unweighted pair‐group means cluster analysis, resulting in 10 clusters corresponding to named variety designations. Similarity among varieties ranged from 0.60 to 0.72. Diversity within varieties was identified. Similarity within named varieties (accessions with the same varietal name) ranged from 0.75 to 0.96. Differences in several markers were found among 34 accessions. Intravarietal diversity was shown to exist within the Spanish olive varieties grown in the region surrounding Valencia.