Deletion of the GI-2 integrase and the wbkA flanking transposase improves the stability of Brucella melitensis Rev 1 vaccine

Brucella melitensis Rev 1 is the best vaccine available for the prophylaxis of small ruminant brucellosis and, indirectly, for reducing human brucellosis. However, Rev 1 shows anomalously high rates of spontaneous dissociation from smooth (S) to rough (R) bacteria, the latter being inefficacious as vaccines. This S-R instability results from the loss of the O-polysaccharide. To overcome this problem, we investigated whether some recently described mechanisms promoting mutations in O-polysaccharide genes were involved in Rev 1 S-R dissociation. We found that a proportion of Rev 1 R mutants result from genome rearrangements affecting the wbo O-polysaccharide loci of genomic island GI-2 and the wbkA O-polysaccharide glycosyltransferase gene of the wbk region. Accordingly, we mutated the GI-2 int gene and the wbk IS transposase involved in those arrangements, and found that these Rev 1 mutants maintained the S phenotype and showed lower dissociation levels. Combining these two mutations resulted in a strain (Rev 2) displaying a 95% decrease in dissociation with respect to parental Rev 1 under conditions promoting dissociation. Rev 2 did not differ from Rev 1 in the characteristics used in Rev 1 typing (growth rate, colonial size, reactivity with O-polysaccharide antibodies, phage, dye and antibiotic susceptibility). Moreover, Rev 2 and Rev 1 showed similar attenuation and afforded similar protection in the mouse model of brucellosis vaccines. We conclude that mutations targeting genes and DNA sequences involved in spontaneous O-polysaccharide loss enhance the stability of a critical vaccine phenotype and complement the empirical stabilization precautions taken during S Brucella vaccine production.     

Comparative Analysis of Native <Emphasis Type="Italic">Crocus</Emphasis> Taxa as a Great Source of Flavonoids with High Antioxidant Activity

Native Crocus taxa are abundant in regions with moderate and dry climate; however, their potential in the food industry has thus far been neglected. The objective of this study was to compare the quantitative and qualitative flavonoid content and antioxidant and cytotoxic activity in organs of several populations of Croatian native taxa and assess their potential for application in the food industry. The dominant flavonoids in the native Crocus taxa were kaempferol and quercetin. Tepals contained similar or higher concentrations of flavonoid-glycosides than other organs. Tepals from Cvv1 and Cvv2 populations contained more flavonoid-glycosides than the commonly used spice saffron (C. sativus stigmas). The FRAP antioxidant activity of Cvv1 and Cvv5 tepals was similar to that of standard Trolox. DPPH inhibition of Cvv1 and Cvv3 tepals was within the range of that recorded for saffron. Cvv1 tepals significantly reduced reactive oxygen species in the broadest concentration range (50–1000 μg/ml), and showed considerable antioxidant activity in the ABTS assay, equal to 82% of standard Trolox antioxidant activity. A significantly higher concentration of kaempferol-rutinoside was recorded in this than in other taxa. The flavonoid showed a very strong or strong correlation with antioxidant assays results, and a negative correlation with cellular reactive oxygen species concentration. We therefore presumed that kaempferol-rutinoside is one of the main antioxidant phenolics in Crocus tepals. None of the tested extracts showed cytotoxicity toward Caco-2 cells. The results revealed that Cvv tepals have potential as a food supplement and are a promising material for further food safety tests.     

The impact of dry and wet cleaning on fabric electromagnetic field shield effect

The aim of this research is to investigate shield effect properties of the fabrics with inox yarns included in the construction, which are used for special professional garments and should protect from electromagnetic microwave irradiation. The investigation was done prior to and after the professional care procedures of dry and wet cleaning in ten cleaning cycles. Shield effect measurements were done on the face and on the back of the fabric, weftwise and warpwise, prior to and after the first, third, fifth, seventh and tenth cycles of dry and wet cleaning, at the frequencies from 0.9 to 2.4 GHz. The results obtained indicated that shield effect of the fabric tested was reduced after professional care procedures, especially so after 5 cycles. The investigations also revealed that shield effect could be considerably enhanced if the inox yarns were incorporated into the fabric in the direction of the warp.     

Integrative approach of histopathology and histomorphometry of common carp (Cyprinus carpio L.) organs as a marker of general fish health state in pond culture

The semi‐intensive system of common carp monoculture in earthen ponds is currently the most practiced fish rearing system in several countries in Eastern Europe. This system is based on the addition of supplemented feed to ponds to achieve faster growth. In this study, the general state of the fish health was monitored when three different types of feeds were used: grain cereals (CF), commercial pelleted feed and extruded feed. The markers used were: (i) the histopathological index (HP) of the main organs: gills, liver and kidney; and (ii) the histomorphometric parameters of the hepatocytes and enterocytes to assess the state of the digestive system in fish fed different types of supplemented feed. At the end of the experiment, the fish were in good health, since irreversible lesions were not noticed to a greater extent in any of the fish organs. There were also no other severe lesions present. However, the results showed differences in the HP indices of the liver and gills, as well as the total HP index, indicating a better health status of the fish in the CF group. The histomorphometric results showed larger cytoplasmic areas of hepatocytes in fish fed CF, which is attributed to glycogen accumulation and higher numbers of goblet cells in the distal intestines of fish fed compound feeds. This can be attributed to slight inflammation.     

柑橘属2n花粉自然发生与沙田柚2n花粉诱导研究

果树2n配子育种较其他植物起步晚,尤其是利用2n花粉进行育种的研究在柑橘上还无先例。试验中初步观察了柑橘属部分品种的2n花粉自然发生情况,证实了柑橘属内未减数花粉的存在。同时通过秋水仙碱不同浓度与作用时间处理促进了柑橘2n花粉的发生,将长寿沙田柚2n花粉发生比例由当年的自然发生率0.87%提高到9.76%。初步观察了2n花粉离体萌发状况,其萌发率与花粉管伸长速度较为正常,但萌发启动比单倍性花粉慢。     

The Characterization of Heavy Metals in the Grapevine (Vitis vinifera) Cultivar Rkatsiteli and Wild Blackberry (Rubus fruticosus) from East Serbia by ICP-OES and BAFs

Inductively coupled plasma optical emission spectroscopy, in combination with calculation of various biological factors, was used in this study not only for a simple illustration of the present status of some essential and non-essential metals such as copper (Cu), zinc (Zn), nickel (Ni), lead (Pb), arsenic (As), and cadmium (Cd) in the grapevine cultivar Rkatsiteli and wild blackberry from East Serbia but also for the estimation of plant bioaccumulation potentials. Great majority of detected metal concentrations were in normal ranges except in few cases: Zn concentrations were at the level of deficiency in almost all plant parts, and some Cu and As concentrations were at the levels that could be considered phytotoxic. Biological accumulation factors provided very informative data on metal accumulation and translocation in both plants and pointed to low accumulation rates of metals except in the case of Cu, Zn, and As in some wild blackberry tissues.     

Biological Activities of Aqueous and Organic Extracts from Tropical Marine Sponges

We report on screening tests of 66 extracts obtained from 35 marine sponge species from the Caribbean Sea (Curaçao) and from eight species from the Great Barrier Reef (Lizard Island). Extracts were prepared in aqueous and organic solvents and were tested for hemolytic, hemagglutinating, antibacterial and anti-acetylcholinesterase (AChE) activities, as well as their ability to inhibit or activate cell protein phosphatase 1 (PP1). The most interesting activities were obtained from extracts of Ircinia felix, Pandaros acanthifolium, Topsentia ophiraphidites, Verongula rigida and Neofibularia nolitangere. Aqueous and organic extracts of I. felix and V. rigida showed strong antibacterial activity. Topsentia aqueous and some organic extracts were strongly hemolytic, as were all organic extracts from I. felix. The strongest hemolytic activity was observed in aqueous extracts from P. acanthifolium. Organic extracts of N. nolitangere and I. felix inhibited PP1. The aqueous extract from Myrmekioderma styx possessed the strongest hemagglutinating activity, whilst AChE inhibiting activity was found only in a few sponges and was generally weak, except in the methanolic extract of T. ophiraphidites.     

荷斯坦育肥牛饮喂甲醛液的增重效果试验

选用西宁市奶牛场荷斯坦育肥牛 1 6头 ,随机分为试验组和对照组 ,每组为 8头。试验组饮喂 1 50 ppm甲醛液 ,对照组饮自来水。试验期 50天。结果表明 ,试验组和对照组平均增重分别为 51 .0 0千克和 2 8.50千克 ,日增重分别为 1 0 2 0克和 570克 ,差异极显著 (p<0 .0 1 )。     

In vitro determination of the indigestible fraction in foods: an alternative to dietary fiber analysis

Dietary fiber (DF) intakes in Western countries only accounts for about one-third of the substrates required for colonic bacterial cell turnover. There is a general trend among nutritionists to extend the DF concept to include all food constituents reaching the colon. In this line, a method to quantify the major nondigestible components in plant foods, namely, the indigestible fraction (IF), is presented. Analytical conditions for IF determination are close to physiological. Samples, analyzed as eaten, were successively incubated with pepsin and alpha-amylase; after centrifugation and dialysis, insoluble and soluble IFs were obtained. IF values include DF, resistant starch, resistant protein, and other associated compounds. IF contents determined in common foods (cereals, legumes, vegetables, and fruits) were higher than DF contents. Calculated IF intakes were close to the estimated amount of substrates reaching the colon. IF data could be more useful than DF data from a nutritional point of view; therefore, IF is proposed as an alternative to DF for food labeling and food composition tables.