Detection and localization of oxidized proteins in muscle cells by fluorescence microscopy

In meat, no detailed studies on the intracellular distribution of oxidized proteins during oxidative stress have been performed, to our knowledge. Therefore, we used fluorescence microscopy to detect and locate protein carbonyls, oxidation products of basic amino acids, generated in bovine M. Rectus abdominis during either exposition to a chemical free radical generating system, or refrigerated storage, or cooking. The technique consisted of an immunohistochemical detection of carbonyls by reaction with the specific probe DNPH (2,4-dinitrophenylhydrazine) followed by the sequential addition of a first antibody against DNPH-carbonylated proteins and a CY3-labeled secondary antibody. The fluorescence of the CY3 probe increased regularly with level of free radical generating system and storage time. Moreover, an important heterogeneity of carbonyl distribution was observed, with a higher oxidation level at the periphery than inside the muscle cells. Cooking induced fluorescence increase only at the periphery of cells. Specific coloration of collagen by Sirius red showed that collagen was not involved in fluorescence. We can deduce that accumulation of oxidized proteins observed in the cell periphery was linked to membrane protein oxidation and not to connective tissue oxidation. Biochemical assays were performed in parallel on membrane and myofibrillar proteins to provide complementary quantitative data on level of oxidized proteins.     

Ecological benefits provided by alley cropping systems for production of woody biomass in the temperate region: a review

In temperate Europe alley cropping systems which integrate strips of short rotation coppices into conventional agricultural fields (ACS) are receiving increasing attention. These systems can be used for crops and woody biomass production at the same time, enabling farmers to diversify the provision of market goods. Adding trees into the agricultural land creates various additional benefits for the farmer and society, also known as ecosystem services. However, tree-crop interactions in the temperate region have not been adequately substantiated which is identified as a drawback to the practical implementation of such systems. In order to bridge this gap, the current paper aims to present a comprehensive overview of selected ecosystem services provided by agroforestry with focus on ACS in the temperate region. The literature indicates that compared with conventional agriculture ACS have the potential to increase carbon sequestration, improve soil fertility and generally optimize the utilization of resources. Furthermore, due to their structural flexibility, ACS may help to regulate water quality, enhance biodiversity, and increase the overall productivity. ACS are shown as suitable land use systems especially for marginal sites. Based on the available data collected, we conclude that ACS are advantageous compared to conventional agriculture in many aspects, and therefore suggest that they should be implemented at a larger scale in temperate regions.     

Management of paddlefish fry and juveniles in Bulgarian conditions

When paddlefish larvae were reared in tanks and fed on zooplankton, a length of 12–15 cm and a weight of 9.2 g had been obtained by the second month after hatching. After a further 2-months of pond rearing, the survival rate was 48%, and the juveniles had reached an average weight of 27.74 g and length of 22.5 cm. Much better growth rate was achieved when both paddlefish fry and juveniles were reared only on artificial food in tanks. At the end of the first month, a weight of 11.6 g had been reached; at the second month the paddlefish weight was 33.29 g, at the third month it was 60.2 g, and at the fourth month it was 127.7 g. There were two cases in which, respectively, 1,000 and 1,500 paddlefish with an average body weight of 33–60 g were stocked in a reservoir. At the age 5 months the fish had reached, or exceeded, an average weight of 500 g, and, after 1 year, it was over 700 g, which is an indicator of the good rearing opportunities this reservoir offered.     

Evaluation of two antimicrobial therapies in the treatment of Leptospira borgpetersenii serovar hardjo infection in experimentally infected cattle

This study demonstrated the ability of the antimicrobials tulathromycin (Draxxin) and ceftiofur crystalline free acid sterile suspension (Excede) to clear the spirochete Leptospira borgpetersenii serovar hardjo type hardjo-bovis (L. hardjo-bovis) from experimentally infected cattle. Treatment with tulathromycin resulted in clearance of L. hardjo-bovis organisms from the urine and kidney tissue of all animals (9 of 9), and treatment with ceftiofur crystalline free acid resulted in clearance of the organisms from the urine of 8 of 10 heifers and the kidney tissue of all 10 animals. In contrast, 10 of 10 placebo-treated cattle had L. hardjo-bovis organisms in their urine and 8 of 10 had the organisms in kidney tissue.     

Effect of meat cooking on physicochemical state and in vitro digestibility of myofibrillar proteins

The effect of meat cooking was measured on myofibrillar proteins from bovine M. Rectus abdominis. The heating treatment involved two temperatures (100 degrees C during 5, 15, 30, and 45 min and 270 degrees C during 1 min). Protein oxidation induced by cooking was evaluated by the level of carbonyl and free thiol groups. Structural modifications of proteins were assessed by the measurement of their surface hydrophobicity and by their aggregation state. With the aim of evaluating the impact of heat treatment on the digestive process, myofibrillar proteins were then exposed to proteases of the digestive tract (pepsin, trypsin, and alpha-chymotrypsin) in conditions of pH and temperature that simulate stomach and duodenal digestion. Meat cooking affected myofibrillar protein susceptibility to proteases, with increased or decreased rates, depending on the nature of the protease and the time/temperature parameters. Results showed a direct and quantitative relationship between protein carbonylation (p<0.01) and aggregation (p<0.05) induced by cooking and proteolytic susceptibility to pepsin. However, no such correlations have been observed with trypsin and alpha-chymotrypsin.     

How is agroforestry perceived in Europe? An assessment of positive and negative aspects by stakeholders

Whilst the benefits of agroforestry are widely recognised in tropical latitudes few studies have assessed how agroforestry is perceived in temperate latitudes. This study evaluates how stakeholders and key actors including farmers, landowners, agricultural advisors, researchers and environmentalists perceive the implementation and expansion of agroforestry in Europe. Meetings were held with 30 stakeholder groups covering different agroforestry systems in 2014 in eleven EU countries (Denmark, France, Germany, Greece, Hungary, Italy, Netherlands, Portugal, Spain, Sweden and the United Kingdom). In total 344 valid responses were received to a questionnaire where stakeholders were asked to rank the positive and negative aspects of implementing agroforestry in their region. Improved biodiversity and wildlife habitats, animal health and welfare, and landscape aesthetics were seen as the main positive aspects of agroforestry. By contrast, increased labour, complexity of work, management costs and administrative burden were seen as the most important negative aspects. Overall, improving the environmental value of agriculture was seen as the main benefit of agroforestry, whilst management and socio-economic issues were seen as the greatest barriers. The great variability in the opportunities and barriers of the systems suggests enhanced adoption of agroforestry across Europe will be most likely to occur with specific initiatives for each type of system.     

Differentiation of <Emphasis Type="Italic">Erwinia amylovora</Emphasis> strains from Bulgaria by PCR-RFLP analysis

Fifty strains of Erwinia amylovora isolated in Bulgaria from different host plants and locations as well as in different years were analysed by RFLP analysis of the pEA29 PstI amplified fragment with HpaII. All the strains formed three well-resolved fragments (large—from 365 to 440 bp, medium—about 341 bp and small—about 180 bp).The strains were classified into three RFLP groups based on the polymorphism in the length of the largest fragment. This fragment was of intermediate size for 63% of the strains, and it was the longest (from 410 to 440 bp) for 29% of the strains. The variable region was sequenced for five strains. The DNA sequence analysis confirmed the different size of the largest fragment. Ten or more than ten SSRs were found for the strains in the group with the largest size of the largest fragment. Some correlation between the RFLP profiles and the origin of the strains was revealed. The RFLP profiles displayed stability in certain strains isolated from the same trees and orchards, but in different years. The number of SSRs was different in strains isolated from one and the same host plant, orchard and year, and also in strains isolated from the same host plant and orchard, but in different years. This could indicate that under natural conditions the fire blight symptoms might be caused by a mixture of E. amylovora strains with different SSR numbers, and so coexistence of distinguishable strains or a change in the population could be assumed.     

Investigation on fecundity, follicles and free embryo size of pond-reared pike (Esox lucius) of different age and size

The purpose of the present study is to investigate the gonadosomatic index (GSI %), the absolute and relative fecundity of one-year-old pike weighing over 400 g and at the same time to study the dependence between the egg size and the size, behavior and vitality of the free embryos obtained from one- and two-year-old spawners. The study involved two weight groups of females, differing in age and body weight and length—one-year-old (W = 514 g, SL = 36.1 cm) and two-year-old matured pike (W = 1454 g, SL = 49.3 cm). Ovary samples were fixed and egg follicles containing maturing oocytes counted and weight. The weight and length of the free embryos from semi-artificial spawning were measured. The results showed that, when raising this species under farmed conditions, more than 40% (in rarer cases 90%) of one-year-old pike females reached over 400 g and 35 cm (SL) and reached puberty. Absolute fecundity of 15,030 follicles (30 follicles per gram body weight) was observed; the GSI was nearly 15% and the follicle weight reached 3.7 mg. This data differs significantly from that obtained from the larger two-year-old fishes: absolute fecundity 41,363 follicles (28 follicles per gram body weight), GSI nearly 20%, follicle weight 5.8 mg. Results showed that the different follicle size determines the free-embryo size. A positive linear correlation was found between the egg follicle weight and the free-embryos weight (r = 0.7143). The free embryos obtain from the one- and two-year-old spawners differed significantly both in terms of their weight (7.13 mg against 10.61 mg) and total length (0.81 cm against 0.97 cm), the differences being 1.5- and 1.2-fold, respectively.