Metabolite profiling of wheat grains (Triticum aestivum L.) from organic and conventional agriculture

In some European community countries up to 8% of the agricultural area is managed organically. The aim was to obtain a metabolite profile for wheat (Triticum aestivum L.) grains grown under comparable organic and conventional conditions. These conditions cannot be found in plant material originating from different farms or from products purchased in supermarkets. Wheat grains from a long-term biodynamic, bioorganic, and conventional farming system from the harvest 2003 from Switzerland were analyzed. The presented data show that using a high throughput GC-MS technique, it was possible to determine relative levels of a set of 52 different metabolites including amino acids, organic acids, sugars, sugar alcohols, sugar phosphates, and nucleotides from wheat grains. Within the metabolites from all field trials, there was at the most a 50% reduction comparing highest and lowest mean values. The statistical analysis of the data shows that the metabolite status of the wheat grain from organic and mineralic farming did not differ in concentrations of 44 metabolites. This result indicates no impact or a small impact of the different farming systems. In consequence, we did not detect extreme differences in metabolite composition and quality of wheat grains.     

Crocin bleaching assay (CBA) in structure-radical scavenging activity studies of selected phenolic compounds

The applicability of the crocin bleaching assay (CBA) to structure-activity relationship (SAR) studies of a great number (n = 39) of selected phenolic compounds was thoroughly investigated. The focus was on the activity of hydroxybenzoic, hydroxyphenylacetic, hydroxyphenylpropanoic, and hydroxycinnamic acids. Other assays [oxygen radical absorbance capacity (ORAC), lipid oxidation] were applied when necessary. Hydroxybenzoic acids were less active than the respective simple phenols. The position of the -COOH group relative to hydroxyl substituents was critical. The number and position of the -OH groups governed the order and size of activity within the subgroup of these acids. Gallic acid was the most active, being 1.6- and 3.4-fold superior to protocatechuic and syringic acids, respectively. The effect of proximity of the -COOH group to the phenyl ring was more distinct for 3,4-guaiacol acids (ferulic > dihydroferulic congruent with homovanillic > vanillic) than for 3,4-catechol ones (caffeic > protocatechuic > or = dihydrocaffeic congruent with homoprotocatechuic). Compounds such as vanillin, tyrosol, ferulic acid derivatives, rosmarinic acid, and quercetin were examined to reinforce discussion on the basis of physical organic chemistry principles. Taking into account the acidity of most compounds, the CBA-derived order of activity was meaningful.     

Endoscopic examination of snakes by access through an air sac

Sixteen boa constrictors (Boa constrictor), three royal pythons (Python regius) and 15 Burmese pythons (Python molurus bivittatus) were examined endoscopically by access through the air sac. The snakes were immobilised in a ventral position using a half-open anaesthetic system with assisted ventilation and a mixture of isoflurane and oxygen. The rigid endoscope was introduced percutaneously and the internal structure of the lungs and the air sac, and the shape, size and external surface of the liver were visible in the cranial direction. In the smaller snakes the bifurcation and caudal part of the trachea could be viewed, provided the endoscope was positioned in a retrograde orientation. The caudal orientation of the endoscope made it possible to view the gall bladder and the size, shape and surface of the spleen. In some cases, the pancreas and the surface of the stomach and colon could be monitored. Endoscopy through the air sac also made it possible to check the major veins in the coelom. The snakes were monitored for at least 30 days after the intervention and no changes in their respiratory function or general health were observed.     

The image of exocytosis during neutrophils and macrophages phagocytic activities in inflammation of mammary gland triggered by experimental Staphylococcus aureus infection

The experiments were carried out in five clinically normal virgin heifers. Before the experimental infection and at 24, 48, 72 and 168 h after the infection, respective mammary glands were rinsed with phosphate buffered saline. Neutrophils as well as macrophages underwent a classic exocytosis accompanied by translocation of lysosomal granules. The granules filled the protuberances of the plasmalemma and after the protuberances separated from the cell, they entered the extracellular space in the shape of round bodies of different sizes. After exocytosis, neutrophils displayed a smaller nucleus/cytoplasm ratio, a greater chromatin density of the nucleus, and an overall smaller size. Macrophages phagocytosed bacteria and/or neutrophils with and without signs of apoptosis (early and late apoptotic respectively) and neutrophils after exocytosis. Macrophages underwent cytolysis that was accompanied by extrusion of granules, phagosomes and phagolysosomes containing phagocytosed bacteria or neutrophils. Confluences were formed in which the process of digestion continued. Apoptosis of neutrophils gradually appeared and intensified in resolution of inflammation. The macrophages contributed to the inactivation of bacterial noxa as well as of histotoxic contents of neutrophils. Nevertheless, macrophages often underwent cytolysis at the site of inflammation.     

Cloning, expression and purification of feline proinsulin

Feline proinsulin was cloned and expressed using a bacterial expression system. It was then purified from inclusion bodies using size exclusion chromatography and further processed including reduction of the protein. Following refolding, proinsulin was purified by reversed-phase high-performance liquid chromatography (RP-HPLC). RP-HPLC and mass spectrometric analysis indicated that the proinsulin contained the correct disulfide bridging pattern. This proinsulin can be used for therapeutic and diagnostic purposes.     

An outbreak of Salmonella Abortusovis abortions in sheep in south Croatia

The aim of the study was to give an account of the epidemic of abortions in sheep caused by Salmonella enterica ssp. enterica serovar Abortusovis, which occurred in Dalmatia, south Croatia, in winter 2003-2004. Five sheep flocks with rate of abortion ranging from 22% to 38% during the last-third of gestation were examined. Salmonella Abortusovis was isolated from 13 vaginal smears and two fetuses. Direct inoculation was found to be superior to pre-enrichment and enrichment in selective broth for Salmonella Abortusovis isolation. The isolates were biochemically identified, and characterized by serotyping and polymerase chain reaction based on the amplification of the IS200 sequence specific for Salmonella Abortusovis. A fragment of 900 bp was detected in all Salmonella Abortusovis isolates. The sensitivity testing of the isolates, carried out by the disk diffusion method and the determination of the minimal inhibitory concentrations, resulted in a high sensitivity to almost all antimicrobials used. Only two isolates were moderately sensitive to oxytetracycline, whereas one isolate showed resistance to streptomycin. Campylobacter fetus ssp. fetus and Listeria monocytogenes were excluded as causative agents of abortion in sheep by culture testing, and brucellosis, leptospirosis, Q fever and chlamydiosis by serological testing.     

Chlamydiales in guinea-pigs and their zoonotic potential

The aim was to detect and characterize chlamydial infections in guinea-pigs (GP) with ocular disease, study their pathogenicity and zoonotic potential and to test for the presence of Acanthamoebae spp. in GP eyes and to investigate whether they could act as vectors for Chlamydia-like organisms. Overall 126 GP, of which 77 were symptomatic, were screened by clinical examination, cytology, gross pathology, histology, immunohistochemistry, polymerase chain reaction (PCR) and bacteriology. A new Chlamydiaceae-specific intergenic spacer rRNA gene PCR, designed to amplify this segment linking the 16S and 23S regions, was performed. DNA samples were also received from one owner including samples of his cat and rabbit. Guinea-pigs: 48 of 75 symptomatic, but only 11 of 48 asymptomatic GP were positive by PCR for Chlamydophila caviae guinea-pig inclusion conjunctivitis (GPIC) (P < 0.0001). Eighteen of 75 or 15/48, respectively, were positive for DNA from Chlamydia-like organisms. Acanthamoebae-DNA could be found in two GP, of which one was symptomatic. Owner, cat and rabbit: Samples of all three species were positive by PCR for C. caviae GPIC and the owner's one-day disposable contact lenses showed a positive PCR result for the Chlamydia-like organism Parachlamydia acanthamoebae. No Acanthamoebae-DNA could be detected. This study is the first to describe Chlamydia-like organisms in GP and to detect C. caviae GPIC in human, cat and rabbit. Therefore, C. caviae GPIC could pose a zoonotic potential. We believe that the finding of C. caviae GPIC in species other than GP is probably not unique.     

Effects of dietary neutral detergent fibre on production performance, nutrient utilization, caecum fermentation and fibrolytic activity in 2- to 3-month-old New Zealand rabbits

An experiment was conducted to determine the effects of different dietary neutral detergent fibre (NDF) levels on growth performance, nutrient utilization, caecum development, caecal fermentation, slaughter performance, immune organ and fibrolytic activity in 2- to 3-month-old New Zealand meat rabbits. Eighty 2-month-old rabbits were allocated in individual cages for five treatments in which they were fed each diet with NDF at 240, 270, 300, 330 and 360 g/kg respectively. The results are as follows: the average daily gain of 300 g/kg NDF group was the highest (30.09 g/day) and higher than that of 240 g/kg NDF group (p < 0.05). The feed/gain ratio of 300 g/kg NDF group was the lowest (4.27) and lower than those of 240 and 360 g/kg NDF groups (p < 0.05). The coefficient of total tract apparent digestibility (CTTAD) of energy, crude protein, crude fibre and ether extract decreased when dietary NDF increased (p < 0.05); CTTAD of NDF increased when dietary NDF increased (p < 0.01). Digestible nitrogen and retained nitrogen of 240, 270, 300 and 330 g/kg NDF groups were higher than that of 360 g/kg NDF group (p < 0.05). The caecum weight, the proportion of caecum weight to body weight and the acetic acid ratio of volatile fatty acid rose when dietary NDF concentration increased (p < 0.05). The NH(3)-N concentration of caecum dropped when dietary NDF increased (p < 0.05). The fibrolytic activity increased when the dietary NDF increased (p < 0.01).     

Treatment of fractured antebrachial bones in both thoracic extremities in toy breed dogs by application of reconstruction plates

Stabilization of fractures at distal third of antebrachial bones in toy breed dogs is often connected with bone healing complications, including delayed union or non-union. In the present study 9 toy breed dogs with fractures at distal third of the epiphysis of two antebrachial bones were treated. Each dog had fractured both thoracic limbs. In all the dogs, internal stabilization was applied using the 1 mm, 4 or 5 -hole reconstructive mini plates. In three dogs mini plates were applied in treatment of non-union of antebrachial bones. In each case radius bone was stabilized only. X-ray examination performed in all the dogs 10 weeks postoperatively showed complete adhesion of treated bones. It was found that application of reconstructive mini plates in treatment of simultaneous fractures of distal shaft of both antebrachial bones is effective therapeutic method.     

The effect of increased ozone concentrations in the air on selected aspects of rat reproduction

Five-month-old male rates were exposed to 0.5 ppm ozone for 50 days, 5 hours a day. A week before the completion of ozone exposure, a biological test was performed to determine the fertilization rate and the survival rate of newborns in both ozone-exposed and control animals. After 50 days, the rats were sacrificed with an overdose of halotane, and testes were collected to assess the morphology and motility of spermatozoa. Neither the morphology of spermatozoa nor motility parameters determined by the CASA (computer-assisted sperm analysis) system showed statistically significant differences between ozone-exposed and control males. The number of successful matings and the survival rate of newborns per litter within one year postpartum were also similar in both groups. However, sperm concentration was by 17% lower in ozone-exposed rats, compared with the control animals.