Moderate ferulate and diferulate levels do not impede maize cell wall degradation by human intestinal microbiota

The degradation of plant fiber by human gut microbiota could be restricted by xylan substitution and cross-linking by ferulate and diferulates, for example, by hindering the association of enzymes such as xylanases with their substrates. To test the influence of feruloylation on cell wall degradability by human intestinal microbiota, nonlignified primary cell walls from maize cell suspensions, containing various degrees of ferulate substitution and diferulate cross-linking, were incubated in nylon bags in vitro with human fecal microbiota. Degradation rates were determined gravimetrically, and the cell walls were analyzed for carbohydrates, ferulate monomers, dehydrodiferulates, dehydrotriferulates, and other minor phenolic constituents. Shifting cell wall concentrations of total ferulates from 1.5 to 15.8 mg/g and those of diferulates from 0.8 to 2.6 mg/g did not alter the release of carbohydrates or the overall degradation of cell walls. After 24 h of fermentation, the degradation of xylans and pectins exceeded 90%, whereas cellulose remained undegraded. The results indicate that low to moderate levels of ferulates and diferulates do not interfere with hydrolysis of nonlignified cell walls by human gut microbiota.     

Evaluation of subgingival bacteria in the dog and susceptibility to commonly used antibiotics

The aim of the present investigation was to evaluate the subgingival aerobic and anaerobic flora of 13 dogs with periodontal disease and the susceptibility of these bacteria to antibiotics currently approved in Italy for treatment of canine infections. Of the anaerobic bacteria, Bacteroides fragilis was most frequently isolated, followed by Peptostreptococcus + Porphyromonas gingivalis and Prevotella intermedia. Of the aerobic bacteria, alpha-hemolytic Streptococcus was most frequently isolated, often associated with Escherichia coli or Pasteurella multocida. Resistance of anaerobic and aerobic bacteria to various antibiotics was generally high. Anaerobic bacteria appeared to be susceptible to amoxicillin + clavulanic acid, doxycycline, and erythromycin; aerobic bacteria appeared to be susceptible to amoxicillin + clavulanic acid, erythromycin, gentamycin, and sulfa-trimethoprim. Bacteroides fragilis was resistant to all of the antibiotics tested. The emerging worldwide problem of bacterial resistance to antibiotics resulting from overuse and misuse of antibiotics is discussed.     

The highly antigenic 53/25 kDa Taenia solium protein fraction with cathepsin-L like activity is present in the oncosphere/cysticercus and induces non-protective IgG antibodies in pigs

Cathepsin L-like proteases are secreted by several parasites including Taenia solium. The mechanism used by T. solium oncospheres to degrade and penetrate the intestine and infect the host is incompletely understood. It is assumed that intestinal degradation is driven by the proteolytic activity of enzymes secreted by the oncosphere. Blocking the proteolytic activity by an antibody response would prevent the oncosphere penetration and further infection. Serine and cysteine proteases including chymotrypsin, trypsin, elastase, and cathepsin L, are secreted by T. solium and Taenia saginata oncospheres when cultured in vitro, being potential vaccine candidates. However, the purification of a sufficient quantity of proteases secreted by oncospheres to conduct a vaccine trial is costly and lengthy. A 53/25 kDa cathepsin L-like fraction partially purified from T. solium cyst fluid was described previously as an important antigen for immunodiagnostics. In this study we found that this antigen is present in the T. solium oncosphere and is also secreted by the cysticercus. This protein fraction was tested for its ability to protect pigs against an oral challenge with T. solium oncospheres in a vaccine trial. IgG antibodies against the 53/25 kDa cathepsin L-like protein fraction were elicited in the vaccinated animals but did not confer protection.     

Correlation of bois noir disease with nettle and vector abundance in northern Italy vineyards

The phytoplasmas associated with bois noir (BN) tuf-type a are transmitted to grapevines by means of Hyalesthes obsoletus Signoret using Urtica dioica L. as the inoculum source. In the period 2003–2008, a research was carried out in six vineyards of northern Italy where a large amount of nettles in the ditches surrounding vineyards was detected. The aim was to establish the nettle control effects on the presence of H. obsoletus and new symptomatic grapevines. PCR and RFLP analyses showed the presence of the BN tuf-type a phytoplasmas in symptomatic grapevines of all vineyards and in the vector. Weed control strategies along ditches were always associated with decreases in nettle coverage and H. obsoletus numbers. A reduction in the percentage of new symptomatic grapevines occurred in all vineyards except for one vineyard where the least reduction in nettle and H. obsoletus was observed. The decrease in H. obsoletus population was significantly correlated with a decrease in nettles along ditches, and the reduction in H. obsoletus population was correlated with a decrease in incidence of new symptomatic grapevines compared with the incidence before the application of weed control measures. Nettle weeding can reduce the incidence of BN tuf-type a, but weed control costs and impacts need to be considered.     

Survey ofPhytophthora infestans populations in Serbia: Mating type, metalaxyl resistance and virulence properties

Summary A total of 37 single-lesion isolates ofPhytophthora infestans were collected during 2000 from different potato growing regions in the central part of Serbia. They were paired in Al and A2 matings to determine their mating types. Of these 37 isolates, 28 were found to belong to the A2 mating type, eight to the Al type and one was found to be self-fertile (A1/A2). Using the in vitro mycelial radial growth method no metalaxyl resistantP. infestans isolate could be found. The rate of occurrence of intermediate isolates was not high, on the average 2.70%. All other isolates were sensitive to metalaxyl. There was no correlation between mating type and the intermediate metalaxyl resistance found.     

Discriminating maize inbred lines using molecular and DUS data

Growing numbers of candidate varieties, decrease of their variability for morphological traits, and internationalization of the national list all contribute to excessive increase of the trial costs, thus creating the need for the improvement of current variety evaluation procedures, especially regards their distinctness, Uniformity, and Stability (DUS) component. Due to rapid advancement in molecular techniques, the use of molecular markers in DUS testing as a complement to, or replacement of, morphological observations became the subject of great interest in scientific studies, and consequently topic for discussion within International Union for the Protection of New Varieties of Plants (UPOV). In order to explore the potential of simple sequence repeat (SSR) markers for distinctness tests, present study involved set of 41 maize inbred lines that were scored for 32 DUS characters prescribed by UPOV and genotyped at 28 SSR loci. Results were largely in favor of the use of molecular markers, revealing or confirming their already known advantages over morphological markers like better consistency with the pedigree, and relatively higher discriminative power. However, their integration into DUS testing protocols still depends upon resolving of several important issues.     

Coffee dietary fiber contents and structural characteristics as influenced by coffee type and technological and brewing procedures

Coffee brews contain considerable amounts of soluble dietary fiber, mainly low substituted galactomannans and type II arabinogalactans. Factors possibly influencing the content and structures of dietary fiber in coffee brews, such as type of coffee, roasting and grinding degree, and brewing procedure, were studied. In addition, several commercial samples such as instant espresso, instant coffee, instant cappuccino, decaffeinated coffees, and coffee pads were analyzed. The dietary fiber contents of the coffee brews ranged from 0.14 to 0.65 g/100 mL (enzymatic-gravimetric methodology), proving an influence of the factors investigated. For example, the drip brew of an arabica coffee contained significantly more soluble dietary fiber than the drip brew of a comparable robusta coffee, and depending on the brewing procedure, the soluble dietary fiber content of beverages obtained from the same coffee sample ranged from 0.26 to 0.38 g/100 mL. Dietary fiber contents of coffee brews were enhanced only up to a certain degree of roast. Drip brews of decaffeinated arabica coffees (commercial samples) contained significantly less dietary fiber than any non-decaffeinated drip brew investigated in this study. The observed differences in the dietary fiber contents were accompanied by changes in the structural characteristics of fiber polysaccharides, such as galactomannan/arabinogalactan ratio, galactose substitution degree of mannans, or galactose/arabinose ratio of arabinogalactans as analyzed by methylation analysis.     

Implementation of immunohistochemistry on frozen ear notch tissue samples in diagnosis of bovine viral diarrhea virus in persistently infected cattle

Background

Bovine viral diarrhea is a contagious disease of domestic and wild ruminants and one of the most economically important diseases in cattle. Bovine viral diarrhea virus belongs to the genus Pestivirus, within the family Flaviviridae. The identification and elimination of the persistently infected animals from herds is the initial step in the control and eradication programs. It is therefore necessary to have reliable methods for diagnosis of bovine viral diarrhea virus. One of those methods is immunohistochemistry. Immunohistochemistry on formalin fixed, paraffin embedded tissue is a routine technique in diagnosis of persistently infected cattle from ear notch tissue samples. However, such technique is inappropriate due to complicated tissue fixation process and it requires more days for preparation. On the contrary, immunohistochemistry on frozen tissue was usually applied on organs from dead animals. In this paper, for the first time, the imunohistochemistry on frozen ear notch tissue samples was described.

Findings

Seventeen ear notch tissue samples were obtained during the period 2008-2009 from persistently infected cattle. Samples were fixed in liquid nitrogen and stored on -20°C until testing. Ear notch tissue samples from all persistently infected cattle showed positive results with good section quality and possibility to determinate type of infected cells.

Conclusions

Although the number of samples was limited, this study indicated that immunohistochemistry on formalin fixed paraffin embedded tissue can be successfully replaced with immunohistochemistry on frozen ear notch tissue samples in diagnosis of persistently infected cattle.     

Detection and structural characterization of thermally generated isoflavone malonylglucoside derivatives

Malonylglucoside isomers were identified by high-performance liquid chromatography with ultraviolet/mass spectrometric detection and nuclear magnetic resonance (NMR). The formation and interconversions of the isomers were monitored in heated buffer and soymilk systems. Two positional or steroisomers of malonylgenistin and malonydaidzin, showing similar UV spectra and molecular weights yet different fragmentation patterns, were detected. NMR characterization of the malonylgenistin isomer revealed its structure to be 4'-O-malonylgenistin, suggesting a malonyl migration from the glucose-6 position to the glucose-4 position. Interconversions of malonylgenistin and its isomer were observed in both buffered and soymilk systems. While malonylgenistin partially converted to the isomer upon thermal treatment, conversion of the isomer to malonylgenistin was more spontaneous. The malonylgenistin isomer represented 6-9% of the total calculated genistein content in soymilk heated at 100 °C for various periods of time. Consequently, disregarding the content of malonylglucoside isomers in processed soy matrices can lead to isoflavone underestimation and misinterpretation of the biological contributions.