Determination of some phenolic acids in Majorana hortensis by capillary electrophoresis with online electrokinetic preconcentration

An online accumulation/mobilization preconcentration technique based on a dynamic pH junction technique and electrokinetic injection was employed for analysis of phenolic acids (sinapic, ferulic, coumarinic, caffeic, syringic, vanillic, and 4-hydroxybenzoic acid) in extracts from Majorana hortensis leaves. Samples were extracted by pressurized solvent extraction with acetone at 150 degrees C and 15 MPa. The capillary electrophoretic method employed 50 mmol.L (-1) sodium borate, pH 9.5, as the sample electrolyte, 50 mmol.L (-1) sodium phosphate, pH 2.5, as the background electrolyte, and 50 mmol.L (-1) sodium phosphate, pH 2.5, with 60 mmol.L (-1) sodium dodecyl sulfate as the mobilization electrolyte. The method allowed 720-fold to 5560-fold preconcentration of the phenolic acids during 30 min of electrokinetic accumulation with detection limits from 0.38 to 4.22 ng.mL (-1).     

Counting elusive animals: Comparing field and genetic census of the entire mountain gorilla population of Bwindi Impenetrable National Park, Uganda

Accurate population size estimates are an essential part of every effective management plan for conserving endangered species. However, censusing rare and elusive wild animals is challenging and often relies on counting indirect signs, such as nests or feces. Despite widespread use, the accuracy of such estimates has rarely been evaluated. Here we compare an estimate of population size derived solely from field data with that obtained from a combination of field and genetic data for the critically endangered population of mountain gorillas (Gorilla beringei beringei) in Bwindi Impenetrable National Park, Uganda. After genotyping DNA from 384 fecal samples at 16 microsatellite loci, the population size estimate was reduced by 10.1% to 302 individuals, compared with 336 gorillas estimated using the traditional nest-count based method alone. We found that both groups and lone silverbacks were double-counted in the field and that individuals constructed multiple nests with an overall rate of 7.8%, resulting in the overestimation of the population size in the absence of genetic data. Since the error associated with the traditional field method exceeded the estimated population growth of 5% in the last 4 years, future genetic censusing will be needed to determine how the population size is changing. This study illustrates that newly improved molecular methods allow fast, efficient and relatively affordable genotyping of several hundred samples, suggesting that genetic censusing can be widely applied to provide accurate and reliable population size estimates for a wide variety of species.     

Residual foot-and-mouth disease virus antibodies in French cattle and sheep six years after the vaccination ban

A serological survey was carried out on French cattle to establish a reference pattern of residual vaccine antibodies and non-specific reactions against the foot-and-mouth disease virus 6 years after the ban on vaccination and in the absence of any foot-and-mouth disease outbreak. Most of the multi-vaccinated cattle still displayed high titres of antibodies and up to 50% of those which had received a single injection still had antibodies. Non-specific reactors were also recorded among animals born during and after 1991. Most of them displayed low titres close to the threshold. Sheep were also tested and, as for cattle, 4.6% of non-specific reactors were recorded, with titres close to the threshold for two-thirds of them. As part of these animals have been resampled and retested, sera revealed negative confirming that these animals are true non-specific reactors. Serological testing as a mean of FMD control should take these facts into account.     

Differentiation of Actinobacillus pleuropneumoniae by PCR-REA based on sequence variability of the apxIVA gene and by ribotyping

During the period of 2001-2003, a total of 591 Actinobacillus pleuropneumoniae field isolates from the Czech Republic were serotyped with a high occurrence of cross-reactions. The cross-reactions were observed in 416 isolates. Most frequently, in 401 isolates (67.9%), cross-reactions with antisera specific for serotypes 9, 11, and/or 1 were observed. Two additional molecular methods, ribotyping and restriction analysis of PCR amplified apxIVA gene (PCR-REA), were therefore used for detailed characterisation of A. pleuropneumoniae. In this subsequent analysis, reference strains representing serotypes 1-12 and 25 field isolates showing the most frequent serotype cross-reactions were examined. PCR-REA enabled all reference strains to be distinguished except for the strains of serotypes 9 and 11. Ribotyping distinguished all reference strains except two pairs of serotypes: 3 versus 6, and 9 versus 11, respectively. Field isolates with serotype cross-reactivity 9, 11, and/or 1 could not be differentiated by either of these methods.     

Spontaneous reduction of intestinal intussusception in five young dogs

Five, 3- to 8-month-old dogs had clinical and ultrasonographic findings consistent with intestinal intussusception, but findings were negative on exploratory celiotomy. Ultrasonography had revealed a target-like mass (median maximal diameter 25 mm) on transverse section and multiple hyperechoic or hypoechoic parallel lines (median length 36.8 mm) on longitudinal section. Blood flow in the intussuscepted bowel was detected in most of the cases. In dogs with similar findings, the authors recommend ultrasonographic reexamination of the abdomen after general anesthesia (but before surgery) to exclude spontaneous reduction of the intussusception.     

Outcome and failure patterns of localized sinonasal lymphoma in cats treated with first‐line single‐modality radiation therapy: A retrospective study

Failure rate and site are not well defined in localized sinonasal lymphoma in cats treated with radiotherapy. In this study, we describe (a) failure pattern, (b) outcome, (c) influence of previously reported prognostic variables on the outcome in cats with suspected localized sinonasal lymphoma. In this multi‐institutional retrospective study, we included 51 cats treated with single‐modality radiotherapy. Cats were irradiated using 10x4.2Gy (n = 32), 12x3Gy (n = 11) or 5x6Gy (n = 8). Regional lymph nodes were prophylactically irradiated in 24/51 cats (47.1%). Twenty‐five cats (49.0%) developed progressive disease: progression was local (nasal) in five (9.8%), locoregional (nodal) in two (3.9%), local and locoregional in three (5.9%), systemic in nine (17.6%) and both local and systemic in six cats (11.8%). No cat receiving prophylactic nodal irradiation had progression in the locoregional lymph nodes. The median time to progression was 974 days (95%CI: 283;1666), with 58% and 53% of cats free of progression at 1 and 2 years, respectively. Median overall survival was 922 days (95%CI: 66;1779) with 61% and 49% alive at 1 and 2 years, respectively. Half of the cats that died of relapse/progression (13/26) died within 6 months of treatment, suggesting possible shortcomings of staging, rapid dissemination of disease or sequential lymphomagenesis. None of the prognostic factors evaluated were predictive of outcome (prednisolone use, anaemia, nasopharyngeal involvement, modified canine Adams tumour stage, protocol, total dose). Radiotherapy is an effective treatment for localized sinonasal lymphoma with a long time to progression. However, in one‐third of the cats, systemic disease progression occurs soon after radiotherapy.     

Production of sexed bovine embryos in vitro can be improved by selection of sperm treatment and co-culture system

The study investigated the effects of sperm sorting, capacitation treatment and co-cultivation on sexed bovine in vitro embryo production. The effect of treatment and co-culture on production of embryos of the preferred sex from unsorted sperm was also studied. Sperm from five breeding bulls was used for fertilization of mature oocytes as follows : Experiment 1, sorted and unsorted sperm (bulls A-E) treated only with heparin in standard co-cultures; Experiment 2, sorted sperm (bulls A-E) treated with heparin-PHE (penicillamine, hypotaurine, and epinephrine) or heparin-caffeine in drop co-cultures; and Experiment 3, unsorted sperm (bull E) treated with either heparin-PHE or heparin-caffeine in both standard and drop co-cultures. In all bulls, treatment with heparin resulted in significantly (p < .05) reduced cleavage and blastocyst rates from sorted sperm, as compared with those from unsorted sperm. In bulls A, B, D and E, treatment of sorted sperm with heparin-PHE in drops significantly increased the blastocyst rate (p < .05). In unsorted sperm of bull E, heparin-PHE treatment in drops resulted in the XX/XY sex ratio inverse to that obtained by heparin-caffeine treatment in standard co-cultures (32.3%/67.7% and 66.7%/33.3%, respectively). In conclusion, the treatment of sorted sperm with heparin-PHE in modified drop co-cultures can be recommended for production of in vitro sexed embryos. The use of unsorted sperm for production of embryos of the preferred sex by selected capacitation treatment and co-culture can be the method of choice in bulls with low IVF yields from sorted sperm.     

Functional Properties of Gluten-Free Pasta Produced from Amaranth, Quinoa and Buckwheat

The use of amaranth, quinoa and buckwheat for the production of gluten-free pasta was investigated in the present study. The aim of the work was to produce pasta of good textural quality, in particular, low cooking loss, optimal cooking weight and texture firmness. The results demonstrated that pasta produced from amaranth had decreased texture firmness and cooking time, while pasta from quinoa mainly showed increased cooking loss. In buckwheat pasta the least negative effects were observed. By combination of all three raw materials to one flour blend in the ratio of 60% buckwheat, 20% amaranth and 20% quinoa, dough matrix was improved. After decreasing dough moisture to 30%, addition of an increased amount of egg white powder of 6% and addition of 1.2% emulsifier (distilled monoglycerides) texture firmness as well as cooking quality of gluten-free pasta produced from such a flour blend reached acceptable values comparable to wheat pasta.     

A point pattern model of the spread of foot-and-mouth disease

The spatial spread of foot-and-mouth disease (FMD) is influenced by several sources of spatial heterogeneity: heterogeneity of the exposure to the virus, heterogeneity of the animal density and heterogeneity of the networks formed by the contacts between farms. A discrete space model assuming that farms can be reduced to points is proposed to handle these different factors. The farm-to-farm process of transmission of the infection is studied using point-pattern methodology. Farm management, commercial exchanges, possible airborne transmission, etc. cannot be explicitly taken into account because of lack of data. These latter factors are introduced via surrogate variables such as herd size and distance between farms. The model is built on the calculation of an infectious potential for each farm.This method has been applied to the study of the 1967-1968 FMD epidemic in UK and allowed us to evaluate the spatial variation of the probability of infection during this epidemic. Maximum likelihood estimation has been conducted conditional on the absence of data concerning the farms which were not infected during the epidemic. Model parameters have then been tested using an approximated conditional-likelihood ratio test. In this case study, results and validation are limited by the lack of data, but this model can easily be extended to include other information such as the effect of wind direction and velocity on airborne spread of the virus or the complex interactions between the locations of farms and the herd size. It can also be applied to other diseases where point approximation is convenient. In the context of an increase of animal density in some areas, the model explicitly incorporates the density and known epidemiological characteristics (e.g. incubation period) in the calculation of the probability of FMD infection. Control measures such as vaccination or slaughter can be simply introduced, respectively, as a reduction of the susceptible population or as a reduction of the source of infection.