Simultaneous determination of six isoflavonoids in commercial Radix Astragali by HPLC-UV

A HPLC-UV method for the quantification of six major isoflavonoids, calycosin 7-O-beta-D-glucoside (1), formononetin 7-O-beta-D-glucoside (2), (6alphaR, 11alphaR) 3-hydroxy-9,10-dimethoxypterocarpan-3-O-beta-D-glucoside (3), 7,2'-dihydroxy-3',4'-dimethoxyisoflavan-7-O-beta-D-glucoside (4), calycosin (5) and formononetin (6), in Radix Astragali (Huangqi) was developed and validated. The method was proven to be sensitive, specific, accurate and precise, as well as effective and easy.     

Assessment of selected antioxidants in tomato pomace subsequent to treatment with the edible oyster mushroom, Pleurotus ostreatus, under solid-state fermentation

Tomato pomace, delignified by the edible oyster mushroom, Pleurotus ostreatus, could be used as a poultry feed ingredient to provide alpha-tocopherol for retardation of lipid oxidation in postmortem meat if the antioxidant were retained in pomace after fungal fermentation. Experiments were conducted to investigate changes in the content of alpha-tocopherol, lycopene, and beta-carotene in tomato pomace after sterilization and treatment with P. ostreatus from 0 to 104 days. alpha-Tocopherol (39.26 to 31.15 microg/g) and lycopene (17.42 to 11.19 microg/g) significantly decreased during sterilization while beta-carotene (42.56 to 35.44 microg/g) did not. The content of carotenoids decreased by day 26 as compared to 0 day for the control and for treated samples. alpha-Tocopherol decreased during fungal fermentation at day 26 as compared to 0 day for the control and increased during the same period for treated samples. By 104 days, only alpha-tocopherol in control pomace was present in a significant amount. The alpha-tocopherol content of mushroom fruit grown in pomace (74.10 microg/g) and in wheat straw (51.36 microg/g) was not significantly different. Tomato pomace could be used as a substrate to successfully grow edible mushrooms; however, the initial level of selected antioxidants and their reduction during fungal fermentation must be considered when delignified pomace is utilized for selected nutrient content in animal feed or products for human consumption.     

Production of a D-glycero-D-manno-heptosyltransferase mutant of Mannheimia haemolytica displaying a veterinary pathogen specific conserved LPS structure; development and functionality of antibodies to this LPS structure

Previous structural studies of the lipopolysaccharides from the veterinary pathogens Mannheimia haemolytica (Mh), Actinobacillus pleuropneumoniae (Ap) and Pasteurella multocida (Pm) had identified a conserved inner core oligosaccharide structure that was present in all strains investigated. In order to examine the potential of this inner core structure as a vaccine, a mutagenesis strategy was adopted to interrupt a D-glycero-D-manno-heptosyltransferase gene (losB) of Mh. This gene encodes the enzyme responsible for the addition of a D-glycero-D-manno-heptose residue, the first residue beyond the conserved inner core, and its inactivation exposed the conserved inner core structure as a terminal unit on the mutant LPS molecule. Subsequent analyses confirmed the targeted structure of the mutant LPS had been obtained, and complementation with losB in trans confirmed that the losB gene encodes an alpha-1,6-D-glycero-D-manno-heptosyltransferase. Monoclonal antibodies raised in mice to this LPS structure were found to recognise LPS and whole-cells of the truncated mutant and wild-type Mh. The antibodies were bactericidal against a wild-type Mh strain and were able to passively protect mice in a model of Mh disease. This illustrates that it is possible to raise functional antibodies against the conserved inner core LPS structure.     

New strategy for the survey of Toxoplasma gondii in meat for human consumption

Monitoring of Toxoplasma infection in animals destined for human consumption is a great challenge for human toxoplasmosis prevention. This study aimed to compare results obtained from a naturally infected population of sheep using different tests and targeting an original matrix: meat samples and muscle fluids collected at the slaughterhouse. A commercial ELISA test was performed on diaphragm fluids from 419 ovine carcasses collected at the slaughterhouse. A MAT (modified agglutination test) was performed on heart fluids obtained from the same animals. In addition, all hearts were bioassayed in mice. Serological test agreement, the relative sensitivity of ELISA MAT and mouse bioassay as well as a correlation between titres and parasite isolation probability were statistically evaluated. The overall agreement (kappa coefficient=0.64) of ELISA on diaphragm fluids and MAT on heart fluids is substantial and subsequently both tests can be used for epidemiological studies. Relative sensitivity was higher for MAT performed on cardiac fluids (90%) than ELISA on diaphragm fluid (61%). For both serological tests, relative sensitivity is lower in lambs younger than 12 months. Relative sensitivity of mouse inoculation was 42%. A significant correlation was obtained between increasing MAT titres and probability to isolate live parasite from the heart. When the fluid titre was higher than 1:16, parasites were isolated in 65% of cases. When it was lower, isolation failed in 95% of the cases. According to our results, cardiac fluids appear to be a relevant matrix for toxoplasmosis survey in meat.     

Estimation of distance related probability of animal movements between holdings and implications for disease spread modeling

Between holding contacts are more common over short distances and this may have implications for the dynamics of disease spread through these contacts. A reliable estimation of how contacts depend on distance is therefore important when modeling livestock diseases. In this study, we have developed a method for analyzing distant dependent contacts and applied it to animal movement data from Sweden. The data were analyzed with two competing models. The first model assumes that contacts arise from a purely distance dependent process. The second is a mixture model and assumes that, in addition, some contacts arise independent of distance. Parameters were estimated with a Bayesian Markov Chain Monte Carlo (MCMC) approach and the model probabilities were compared. We also investigated possible between model differences in predicted contact structures, using a collection of network measures.We found that the mixture model was a much better model for the data analyzed. Also, the network measures showed that the models differed considerably in predictions of contact structures, which is expected to be important for disease spread dynamics. We conclude that a model with contacts being both dependent on, and independent of, distance was preferred for modeling the example animal movement contact data.     

What is your diagnosis? Intracranial mass in a cat

Abstract: A 14‐year‐old female spayed cat was presented with a 3–4‐month history of circling to the left and intermittent head pressing. Neurologic examination findings localized the lesion to the left supratentorial region. Using magnetic resonance imaging, an extra‐axial mass was found on the dorsal aspect of the brain at the level of the frontal and parietal lobes, compressing and displacing the brain ventrally and caudally. Craniectomy was performed and the mass was submitted for cytologic and histopathologic evaluation. Impression smears revealed abundant cholesterol crystals and loose clusters of mildly pleomorphic spindle cells, compatible with a meningioma. The histopathologic diagnosis was meningioma with psammoma bodies and numerous cholesterol clefts. Abundant cholesterol crystals within meningiomas in cats may present a diagnostic challenge when nucleated cells are scant. Other differential diagnoses for abundant cholesterol crystals in an intracranial mass include cholesterol granulomas and keratinizing cysts.     

High levels of spatial heterogeneity in the biodiversity of soil prokaryotes on Signy Island, Antarctica

In a previous study, soil bacterial diversity at environmentally distinct locations on Signy Island was examined using denaturing gradient gel electrophoresis (DGGE) profiling, and a range of chemical variables in soils was determined in order to describe variations between them. The dominant bacterial communities of all locations were found to be significantly different, although higher levels of similarity were observed between locations with similar physico-chemical characteristics, such as at penguin rookeries, seal wallows and vegetated soils. Extending this study, here soil prokaryote biodiversity was compared between 15 distinct locations in order to elucidate any interaction between four general habitat types on Signy Island (South Orkney Islands, maritime Antarctic) and any influence of previous human impacts at these sites. Specific sites were selected to represent the range of different soil environments present and to cover a range of environmental factors present in the maritime Antarctic which are known to influence bacterial community composition in soils elsewhere. A diverse prokaryote community is described, again with the majority of excised and sequenced bands belonging to the Bacteroidetes. Although DGGE profiling identified significant differences in prokaryotic biodiversity between all sampling sites, aggregations of banding patterns were also apparent across the different soil environments examined. Correlations between specific DGGE profiles and 10 selected soil parameters suggested that much of this variation could be explained by differences in the levels of environmental disturbance and soil pH. In particular, a greater proportion of variation in soil bacterial diversity was explained by differences in soil properties at human-disturbed locations than at undisturbed locations, with higher explanatory values by edaphic factors in the former and soil metal content in the later. In general, our data indicate that small-scale variation is an important factor in understanding patterns of prokaryotic distributions in soil habitats in the maritime Antarctic environment.     

Study of Defense-related Gene Expression in Grapevine Leaves and Berries Infected with Botrytis cinerea

Defense responses of grapevine towards Botrytis cinerea were investigated. The expression of genes coding for proteins involved in defense were studied: (a) phenylalanine ammonia-lyase (PAL) and stilbene synthase (StSy), (b) an acidic chitinase (VCH3) and a basic chitinase (VCHIT1b), and (c) a polygalacturonase inhibitor protein (PGIP). Since no PGIP was known in grapevine, a complete cDNA sequence was first characterized by PCR and RACE-PCR amplifications. RNAs isolated from infected leaves and infected berries were analysed by semi-quantitative and real-time RT-PCRs. In infected leaves, the expression of PAL, StSy, PGIP and VCH3 genes occurred 6hours post inoculation (hpi). Increase of VCHIT1b gene expression was delayed (24hpi). Maximum levels of induction of these genes were observed at 48hpi, except for the VCH3 gene (24hpi). Activation of these defense responses was not sufficient to stop B. cinerea spread. In berries, no VCH3 gene expression was detected. Maximum levels of induction were observed in stage 3 (loss of berry colour and abundant production of conidia) for the PAL and PGIP genes, and in stage 4 (shrivelled berry) for the StSy and VCHIT1b genes.     

Identification and characterisation of coding tandem repeat variants in incA gene of Chlamydophila pecorum

Bacteria of the family Chlamydiaceae are obligate intracellular pathogens of human and animals. Chlamydophila pecorum is associated with different pathological conditions in ruminants, swine and koala. To characterize a coding tandem repeat (CTR) identified at the 3' end of incA gene of C. pecorum, 51 strains of different chlamydial species were examined. The CTR were observed in 18 of 18 tested C. pecorum isolates including symptomatic and asymptomatic animals from diverse geographical origins. The CTR were also found in two strains of C. abortus respectively isolated from faeces from a healthy ewe and from a goat belonging to asymptomatic herds, but were absent in C. abortus strains isolated from clinical disease specimens, and in tested strains of C. psittaci, C. caviae, C. felis and C. trachomatis. The number of CTR repeats is variable and encode several motifs that are rich in alanine and proline. The CTR-derived variable structure of incA, which encode the Chlamydiaceae-specific type III secreted inclusion membrane protein, IncA, may be involved in the adaptation of C. pecorum to its environment by allowing it to persist in the host cell.