A cattle genetic linkage map was constructed which covers more than 95 percent of the bovine genome at medium density. Seven hundred and forty six DNA polymorphisms were genotyped in cattle families which comprise 347 individuals in full sibling pedigrees. Seven hundred and three of the loci are linked to at least one other locus. All linkage groups are assigned to chromosomes, and all are orientated with regards to the centromere. There is little overall difference in the lengths of the bull and cow linkage maps although there are individual differences between maps of chromosomes. One hundred and sixty polymorphisms are in or near genes, and the resultant genome-wide comparative analyses indicate that while there is greater conservation of synteny between cattle and humans compared with mice, the conservation of gene order between cattle and humans is much less than would be expected from the conservation of synteny. This map provides a basis for high-resolution mapping of the bovine genome with physical resources such as Yeast and Bacterial Artificial Chromosomes as well as providing the underpinning for the interpolation of information from the Human Genome Project.
Scots pine (Pinus sylvestris) genomic libraries were constructed and screened with oligonucleotides probes (GT)10, (CT)10, and (AT)10. Eight microsatellites were identified from 6000 clones screened. The longest microsatellite stretch found, (CT)9(N)21(AT)24, was amplified from bud and single pollen grain samples. In order to clarify the complex amplification pattern revealed, two PCR products were sequenced. The size differences were caused both by varying repeat numbers of the microsatellite stretches and by differences in other parts of the amplified sequence. This kind of complex molecular basis of microsatellite amplification within a species has been previously reported. Microsatellite sequences were used as PCR primers to detect polymorphisms and to estimate the abundance of microsatellites.
The applicability of single strand conformation polymorphism (SSCP) analysis for major histocompatibility complex (MHC) genotyping in sheep was studied. A panel of Ovar-DRB1 exon 2 'allele fingerprints' was defined. The panel could accelerate DRB1 genotyping of new breeds when already existing sequences are used as references in SSCP analysis. In this study, seven new exon 2 sequences and 19 different alleles in total were detected from 31 animals of Finnish and Russian sheep breeds. Ovar-DRB1*0201 was detected in all the six grey Finnsheep animals included in this study, suggesting reduced MHC diversity within these animals.
The presence of the cytolethal distending toxin B gene (cdtB) was examined in eight Helicobacter sp. flexispira reference strains, Helicobacter trogontum ATCC 700114 T and 12 Finnish porcine H. trogontum strains and canine flexispira isolates. Part of the cdtB gene was amplified by PCR with degenerate primers VAT2 and DHF1, cloned and sequenced. The presence/absence of the cdtB gene as determined by PCR was confirmed by Southern hybridization and toxin production by HeLa cell-line experiments. PCR amplification resulted in approximately 700 bp fragments from Helicobacter sp. flexispira taxa 2 (ATCC 49314), 3 (ATCC 49320) and 8 (ATCC 43880, ATCC 49308, ATCC 43879), from six canine isolates as well as from the control strains Helicobacter bilis and Helicobacter hepaticus. The hybridization patterns of HaeIII-, HindIII-and AseI-digested chromosomal DNA confirmed the results of the PCR experiments. The cdtB-positive strains had effects ranging from weak to strong on HeLa cell cultures. PCR amplification from the reference strains Helicobacter sp. flexispira taxa 1 (ATCC 43968), 4 (ATCC 49310) and 5 (ATCC 43966) and H. trogontum (ATCC 700114 T ), and also six of the Finnish strains, was unsuccessful. No toxic effect on HeLa cells was evident when bacterial suspensions of PCR-negative strains were used for toxicity assay. Our results are in accordance with previous observations that the cdtB gene is not present in all Helicobacter species. Further, the presence/absence of the cdtB gene in Helicobacter sp. flexispira strains was in accordance with recent taxonomic analysis of the same strains, which suggests that it could serve as a useful marker in Helicobacter taxonomy.
Malodors from pig manure storage pits are a problem lacking a cost-efficient solution particularly for small pig (Sus domestica) farms. The objective of this study was to reduce emissions of noxious odorants by changing the conditions in the pig manure to favor an altered microbial community. Sugars (maltose, lactose, and saccharose), carbohydrate-rich waste (maltose syrup and wheat flour), and bacterial amendments (Lactobacillus plantarum and L. amylophilus) were tested for their effect on manure pH, bacterial community, and gaseous emissions. In laboratory experiments, a permanent pH reduction was achieved with all carbohydrates when added to a 5% final concentration. Maltose significantly decreased emissions of sulfur-containing compounds, whereas bacterial amendments had little effect on odorants. Lactobacilli were found in manure receiving carbohydrate amendments alone, but Clostridiales, Bacteroidetes, and Enterobacteriaceae were observed in all treatments (including the control). At the mesocosm (60 L) level, maltose syrup and wheat flour amendments caused clear pH reductions in manure, whereas L. plantarum and L. amylophilus had no additional effect. The addition of maltose syrup and wheat flour to a manure storage pit (600 m(3)) lowered the pH slightly. When the manure was spread onto the fields, the observed reduction in odor was found to be reversible, and the release of malodors was delayed rather than eliminated. We conclude that these methods require further development to produce a reliable technical application.
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