Detection of bacterial contamination and DNA quantification in stored blood units in 2 veterinary hospital blood banks

Blood transfusions in veterinary medicine have become increasingly more common and are now an integral part of lifesaving and advanced treatment in small and large animals. Important risks associated with transfusion of blood products include the transmission of various infectious diseases. Several guidelines suggest what infectious agents to screen for in canine and feline transfusion medicine. However, while the risk of bacterial contamination of blood products during storage and administration has not been documented in veterinary medicine, it has emerged as a cause of morbidity and mortality in human transfusion medicine. Clinical experience shows that the majority of blood component bacterial contaminations are caused by only a few species. Unlike other types of bacteria, psychrotolerant species like Pseudomonas spp. and Serratia spp. can proliferate during the storage of blood units at 4°C from a very low titer at the time of blood collection to a clinically significant level (> 10⁵ CFU/mL) causing clinical sepsis resulting from red blood cell concentrate transfusions in human medicine. The purpose of this report was to describe the detection and quantification procedures applied in 4 cases of bacterial contamination of canine and feline blood units, which suggest the need for further investigations to optimize patients’ safety in veterinary transfusion medicine.     

Effects of the extensive culture system as finishing production strategy on biometric and chemical parameters in rainbow trout

The effect of finishing extensive farming period, to reduce fat content and manipulate the fatty acid profile of fish muscle, was evaluated in rainbow trout. Fish were stocked in an artificial lake, in which fish were fed only on naturally available nutrients with no supply of artificial feed, for different lengths of time from 0 to 120 days. No weight loss was noted during the whole finishing period while total length increased from 228±7 to 269±3 mm and the condition factor decreased from 1.41±0.04 to 0.89±0.02. The total fat content of the fillets decreased considerably from 4.7±0.6% at the beginning to 2.4±0.4% and 0.7±0.2% after 45 and 120 days respectively. Fillet fatty acid composition was affected by the time of stocking in the extensive farm. In contrast to the reduction in C18:1n‐9, C18:2n‐6, total monounsaturated fatty acid and total n‐6 percent values, an increase in the C20:5n‐3, C22:6n‐3, total polyunsaturated fatty acid and total n‐3 percent values was observed. It was shown that while other finishing strategies for salmonids have some disadvantages, the extensive culture system seems to be a potentially useful tool for increasing the general quality of the end product.     

Classification of gilthead sea bream (Sparus aurata) from 1H NMR lipid profiling combined with principal component and linear discriminant analysis

The combination of (1)H NMR fingerprinting of lipids from gilthead sea bream (Sparus aurata) with nonsupervised and supervised multivariate analysis was applied to differentiate wild and farmed fish and to classify farmed specimen according to their areas of production belonging to the Mediterranean basin. Principal component analysis (PCA) applied on processed (1)H NMR profiles made a clear distinction between wild and farmed samples. Linear discriminant analysis (LDA) allowed classification of samples according to the geographic origin, as well as for the wild and farmed status using both PCA scores and NMR data as variables. Variable selection for LDA was achieved with forward selection (stepwise) with a predefined 5% error level. The methods allowed the classification of 100% of the samples according to their wild and farmed status and 85-97% to geographic origin. Probabilistic neural network (PNN) analyses provided complementary means for the successful discrimination among classes investigated.     

Effect of Scleroderma Spore Density and Age on Mycorrhiza Formation and Growth of Containerized Eucalyptus globulus and E. urophylla Seedlings

Greenhouse trials with containerized Eucalyptus globulus and E. urophylla seedlings were made to determine effective inoculum spore densities (Scleroderma cepa from temperate Australia, and S. citrinum from sub-tropical China) and spore storage conditions (fresh spores or spores stored for 5 years at room temperature or at 4 °C; S. albidum, S. areolatum and S. cepa from Western Australia). Inoculation with 10⁶ or 10⁸ spores/seedling increased eucalypt growth by up to 46% in height and 42% in dry weight compared to non-inoculated seedlings at 12 weeks after inoculation. Mycorrhizal formation was poor at 10² spores/seeding. Spores stored at 4 °C for 5 years were as effective in forming mycorrhizas as freshly collected spores when a standard density of 10⁶ spores/seedling was applied. It is recommended that spore densities ≥10⁴ be used for inoculation of containerised eucalypts and that spores be stored at 4 °C until use.     

Post-emergent herbicidal activity of cineole derivatives

Essential oils are being investigated as potential herbicides or to provide leads to new environmentally and socially acceptable herbicides. Novel hydroxy and ester derivatives of 1,8-cineole and 1,4-cineole were synthesised, by chemical and biological methods, and have shown pre-emergence herbicidal activity against annual ryegrass and radish. Effects on post-emergence activity of these derivatives, as well as 1,8-cineole, eucalyptus oil and the carboxylic acids from which the esters were derived, against annual ryegrass and radish, are reported here. Results suggest that reduced root and shoot growth observed in pre-emergence herbicidal bioassays were due to post-emergence activity rather than delayed germination. All tested substances had a dose-dependent, post-emergence herbicidal activity against annual ryegrass and radish with many derivatives showing improved activity relative to 1,8-cineole and high-cineole eucalyptus oil. However, results do not support the postulate that cineole esters would be more active than their respective carboxylic acid and the hydroxy cineole. Phytotoxicity of ester derivatives may be due to metabolic cleavage of the esters to the hydroxy cineole and carboxylic acid within the plant.     

Evaluation of different protein sources in fingerling grey mullet <Emphasis Type="Italic">Mugil cephalus</Emphasis> practical diets

An eight-week experiment was carried out to evaluate the effects of different protein sources (fish and haemoglobin meal, soybean meal and torula yeast), in practical diets, on growth, body composition and gut morphology of fingerling grey mullet (Mugil cephalus). Weight gain (%), SGR, FCR, N retention, PER, PGR, FDR and carcass composition of fish were not significantly affected by the dietary protein source. Fish fed the torula yeast based diet showed reduction in growth performance. Histological examinations performed on the alimentary tract of the fish showed a normal structural pattern in the experimental groups, as fundamental histological and histochemical aspects were similar if compared to the control group. The lower growth performance observed in fish fed a torula yeast based diet may be tentatively correlated with the presence of some detrimental morpho-functional aspects in the gut of these fish if compared to fish fed the other diets. Further studies are necessary to confirm this hypothesis.     

N-linked glycosylation in Campylobacter jejuni and its functional transfer into E. coli

N-linked protein glycosylation is the most abundant posttranslation modification of secretory proteins in eukaryotes. A wide range of functions are attributed to glycan structures covalently linked to asparagine residues within the asparagine-X-serine/threonine consensus sequence (Asn-Xaa-Ser/Thr). We found an N-linked glycosylation system in the bacterium Campylobacter jejuni and demonstrate that a functional N-linked glycosylation pathway could be transferred into Escherichia coli. Although the bacterial N-glycan differs structurally from its eukaryotic counterparts, the cloning of a universal N-linked glycosylation cassette in E. coli opens up the possibility of engineering permutations of recombinant glycan structures for research and industrial applications.     

sunTILL: a TILLING resource for gene function analysis in sunflower

Background

Cultivated sunflower (Helianthus annus L.) is a globally important oilseed crop, subjected to intensive genetic and genomic studies. Although classical mutagenesis has successfully been applied to Helianthus genus in the past, we have developed the first sunflower TILLING resource.

Results

To balance the maximum mutation density with an acceptable plant survival rate, a 'kill curve' analysis was first conducted with different ethylmethanesulfonate (EMS) dosages and different exposure times. According to the germination rate, a treatment with 0.7% EMS for 6 h was chosen. An M₂ progeny of 3,651 fertile plants was obtained. Totally, 4.79% of the whole population showed clear aberrant phenotypes. A microsatellite analysis on a representative sample of the original seed stock and mutant lines confirmed the uniformity of the genetic background of plant material. The TILLING procedure was successfully applied to sunflower genome, initially by a CelI-nuclease mismatch cleavage assay coupled with a DNA-pooling level test. To investigate the efficiency of the mutagenic treatment, a pilot screening was carried out on 1,152 M₂ lines focusing on four genes, three involved in the fatty acid biosynthetic pathway and one for downy mildew resistance. A total of 9 mutant lines were identified and confirmed by sequencing; thereby, the estimated overall mutation frequency for the pilot assay resulted to be 1/475 kb.

Conclusion

A first TILLING population for a high throughput identification of EMS-induced point mutations in sunflower genome has been successfully obtained. This represents a powerful tool to a better understanding of gene function in sunflower.