Experimental and natural infection with the enzootic leukosis virus of cattle

A trial was performed with heifers at the age of six to seven months. The animals were experimentally infected with the lymphocytes of a virus-productive donor. Infection was produced in all the nine cases, as demonstrated by means of the positive syncytial test. As indicated by the results of the trial, the antibodies to the enzootic bovine leucosis virus (BLV) were produced soon after experimental infection. A high sensitivity of the serum-neutralization test and the ELISA method was demonstrated in this connection: by these methods, the antibodies were identified already two to three weeks after experimental infection whereas by the immunodiffusion test they could be detected only after five weeks. Twenty-four animals were exposed to natural contact infection. Within 270 days of the trial, the disease after contact was recorded only in one heifer out of the four that were in close contact with the experimentally infected animals. In this case, as compared with experimental infection, the antibodies were produced much later--after 85 to 93 days. Leucosis was recorded in none of the remaining animals. The reasons why such a favourable result was obtained were the thorough disinfection of the stables after blood collections and the strict observance of the aseptic conditions. The results of experimental infection in three cows were identical with those obtained in young cattle. In the experimentally infected dairy cows, antibodies in milk were determined by the ELISA method. As found, in milk the antibodies to BLV appear two to three weeks later than they do in serum. The ELISA method of BLV antibody detection can be used for the identification of infected animals in herds where enzootic bovine leucosis occurs.     

Mucus synthesis in the goblet cells of the small intestine in experimental infection with the coccidium Isospora suis in piglets

The state of mucus synthesis in the goblet cells of the small intestine was studied in conventional piglets infected with a dose of 200,000 oocytes of the coccidium Isospora suis the first and fifth day after parturition. The synthesis of mucus and its chemical characteristics undergo significant changes during the third and fourth day after infection. The activity of acid and neutral mucous substances declines; their level and the physiological synthetic function of goblet cells begin to return to the normal during the period starting on the eight to tenth day after infection. However, there were no fully functioning goblet cells in the broken numerical ratio even at the end of the period of investigation, i.e. the 13th day after infection. The thin surface layer of mucus remained almost unchanged within the whole extent of the small intestine parts studied.     

Fertilizing capacity of the ejaculate of nutria (Myocastor coypus) after the removal of the seminal vesicles as evaluated by the penetration test and natural mating

The fertility of male coypu sperm following seminal vesicle extirpation was investigated using the penetration test into the egg of Syrian golden hamster (Mesocricetus auratus). Ejaculates were obtained from five males by means of electro-ejaculation under halothane narcosis. The results of the zona-free hamster eggs (ZFHE) penetration test showed that the ejaculates of all the surgically treated coypu males were fertile and that ZFHE value fluctuated from 54 to 76.6%. The results obtained in experiments with natural mating revealed that the extirpation of male coypu seminal vesicles did not affect their fertility. In total 47 foetuses were found post mortem in ten coypu females covered by surgically treated males, which on average represented 4.7 foetuses per female.     

Development of specific PCR primers for identification and detection of <Emphasis Type="Italic">Phytophthora capsici</Emphasis> Leon

A PCR-based method was developed for the identification and detection of Phytophthora capsici in pepper plants. Three PCR primers (CAPFW, CAPRV1 and CAPRV2) specific for P. capsiciwere designed based on the sequence of its internal transcribed spacer regions. CAPFW/CAPRV1 amplify a 452 bp product from P. capsici DNA whereas CAPFW/CAPRV2 a 595 bp fragment; neither set amplifies DNA from pepper or several fungi pathogenic to pepper. In conventional (single-round) PCR, the limit of detection was 5 pg DNA for both primer sets, whereas in nested PCR the detection limit for both was of 0.5 fg. However, when the dilution series of target DNA were spiked with plant DNA, amplification declined two-fold in both conventional and nested PCR. The CAPFW/CAPRV2 set in conventional PCR was used to detect P. capsici DNA in inoculated plants. Detection occurred as soon as 8h post-inoculation in stem samples from infected but still symptomless plants. The method was also tested to detect fungal DNA in infected soils.     

Analysis of a site x year experiment with timothy polycross progeny

Summary Results from a cooperative breeding programme with timothy for the northern areas of Scandinavia are presented. The main aim of the programme was to identify genotypes for synthetic populations that are high yielding, adapted over the whole area and stable over years. A polycross comprising 12 genotypes from each of five sites within the region was formed. The subsequent 60 half-sib families and four reference varieties were then compared under sward conditions at the same sites. The trials lasted for three years and results for total dry matter yields are presented.Significant differences in yield between lines were found. There were also significant two and three factor interactions between lines, sites and years. A new two-step procedure is presented to select the genotypes to make up a synthetic variety. Firstly, a superiority measure (Q-value) is used to select a group of genotypes that are high yielding and stable over sites and years. Secondly, the GxE part of the Q-value is split into two terms. One measures adaptation to predictable environmental conditions. The other one measures stability to unpredictable changes in the environment. A plot of these two measures provides a tool for discarding unstable or poorly adapted genotypes. The statistical properties of the selection parameters are discussed.     

Hydraulic Retention Time Influence on Improving Flocculation in the Activated Sludge Processes Through Polyelectrolytes

Hydraulic retention time (HRT) influence improving sludge flocculation with adding the polyelectrolytes (non-ionic, anionic, and cationic) was studied on an activated sludge (AS) system fed with synthetic domestic wastewater (SDW), dairy industry wastewater (DIW), and caramel industry wastewater (CIW). The sludge volumetric index, food/microorganism ratio (F/M), and mixed liquor volatile suspended solids at different HRTs (6, 8 and 10 h) were monitored on an experimental model. Results showed that both SDW and IW had the best sludge flocculation conditions at 8 h and 100 mL of non-ionic polyelectrolyte (0.2 mg L^?1). In addition, this phenomenon reached the organic matter removal efficiencies of 95.9, 95.7, and 94.2% for SDW, DIW, and CIW, respectively. Therefore, optimum HRT increased the organic matter removal efficiencies by 10%, sludge concentration by 37% (22–55%), and F/M ratio by 70%. Moreover, the polyelectrolytes used in AS improved the sludge flocculation by 2.9 times.     

Competitive and allelopathic interference between soybean crop and annual wormwood (Artemisia annua L.) under field conditions

Annual wormwood interference on soybean crop growth and yield may result from competition and allelopathy, which are modulated by crop management. Allelochemicals released by annual wormwood (e.g. artemisinin) may affect the crop directly or indirectly through the effect on the nitrogen fixing symbiont, Bradyrhizobium japonicum. The objectives were (i) to quantify the crop response (i.e. biomass production, nodulation and yield) to weed interference and (ii) to determinate the relative change of competition and allelopathy interferences, when a sublethal dose of herbicide is applied. Two split plot field experiments with three replications were used. The experiment involved a factorial combination of five weed–crop density (soybean/annual wormwood, plants m^?2) levels: D1, pure soybean, 40/0 plants m^?2; D2, 40/2 plants m^?2; D3, 40/4 plants m^?2 and D4, 40/8 plants m^?2, and D5, pure annual wormwood, 0/8 plants m^?2, two activated carbon (allelopathy) levels: C?, with activated carbon (reduced allelopathy) and C+, non activated carbon applied (with allelopathy) and two herbicide levels: H?, untreated and H+, treated with a sub-lethal dose of glyphosate. Activated carbon to adsorb allelochemicals (with and without activated carbon) and glyphosate application (with and no herbicide) were assigned to sub-plots. Increasing weed density did not affect crop biomass at flowering, but changed nodule number and soybean yield with a different pattern depending on carbon and herbicide treatment. Relative crop yield decreased with increasing relative weed biomass. This decrease was particularly drastic when allelopathy was reduced by activated carbon and without herbicide application. The maximum yield losses of 33% in 2006 and 17% in 2007 were observed with the highest weed density (8 plants m^?2). In contrast, without carbon (high allelopathy level), soybean yield remained stable within the explored range of annual wormwood biomass, despite the fact that weed biomass at high densities (D4) was high enough to generate competition. The lack of response to increasing weed density could be related to the indirect effect of allelochemicals interacting with soil microorganisms (i.e. B. japonicum) that positively affected the nodulation (e.g. larger nodules in 2006 and increased nodules biomass due to higher number of roots in 2007 at high densities). With herbicide application, soybean yield of both carbon treatments remained stable when biomass of annual wormwood increased. This research provided strong evidence in support of the existence of positive effect of allelopathic and competitive interactions between annual wormwood and soybean crop under field conditions that may be overridden under herbicide application.     

Morphological characterization of the IVIA persimmon (Diospyros kaki Thunb.) germplasm collection by multivariate analysis

Diospyros kaki Thunb. originated in Eastern Asia, as evidenced of its culture in China as early as several centuries B.C. In the seventh century, persimmon was introduced to Japan and later, in the fourteenth century to Korea. There is no information about persimmon culture in Europe until the seventeenth century, with the spread over the world occurring in the eighteenth century. The genus Diospyros contains more than 400 species, with levels of ploidy ranging from diploid (2n = 2x = 30) up to nonaploid (2n = 9x = 135). The primary economic crop species is Diospyros kaki Thunb., which is mainly hexaploid (2n = 6x = 90) and includes hundreds of cultivars. Although a relatively recent introduction in Europe, the species has adapted well, and the genetic diversity have been expanded with culture and selection for the past 200 years in the Mediterranean basin. These locally adapted cultivars were evaluated with cultivars from Asian origin in a germplasm collection established at IVIA in Valencia, Spain. In this paper 27 cultivars from the IVIA collection were studied by multivariate analysis, and 37 variables were analyzed using a Principal Components Analysis and cluster analysis following the method UPGMA. Studies on correlations and significance among variables identified the most relevant ones, and thus provided information for a future core collection.     

Contamination of soil with Toxocara eggs in urban (Prague) and rural areas in the Czech Republic

Contamination of soil with feline and canine ascarid eggs in public parks, backyards and sand pits in Prague, Czech Republic was investigated in this work. Soil samples from shelters and rural areas were also collected. The comparison of soil from different areas (urban, rural, backyards and shelters) exhibited significant difference (chi(2)=32.16, d.f.=3 and p<0.0001). The highest rate of contamination (45%) was found in backyards inhabited by feral cats. The eggs of Toxocara spp. were found in 20.4% of parks, 10% of shelters and 5% of rural samples. Mean egg density per sample from Prague parks was 6.2 eggs/100g of soil. In 126 composite samples from children's and pits, the prevalence of Toxocara eggs was 11.90%. The number of eggs in positive samples varied from 2 to 22 (per 100g). A high proportion (46.9%) of eggs was fully embryonated. There was no difference between the sand pits with or without formal exclusion of dogs (chi(2)=0.6, d.f.=1 and p<0.0001).     

Thin layer-chromatographic separation of fat soluble and water soluble food dyes

Summary The use of fat soluble and water soluble food dyes in food technology requires reliable proofs and precise identification of these substances in foodstuffs. This problematic has been studied more detailed in our Institute. During these studies some suitable procedures for thin layer chromatographic separation of these substances have been developed.For the separation of fat soluble food dyes the chromatography on thin layers of aluminium oxide and the chromatography on thin layers of starch with reversed phases were used. In the first case good separation was achieved with a solvent system containing petroleum ether and carbon tetrachlorid and their mixtures. In the second case the starch plates were impregnated with paraffin or vegetable oil, and as mobile phases a solvent system containing methanol-water-acetic adic (16 : 3 : 1) and others solvent mixtures currently used in paper chromatography were used.Water soluble food dyes were separated on thin layers of polyamide powder, which was used also for their quantitative isolation from foodstuffs. The best results were achieved with basic solvent system, for example, with a mixture containing ammonia-methanol water (5 : 15 : 80).The developed analytical procedures are suitable for the proof and the separation of fat soluble and water soluble food dyes in all foodstuffs.

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Zusammenfassung Die Verwendung von wasser- und fettlöslichen Lebensmittelfarben erfordert verläßlichen Nachweis und genaue Identifikation dieser Stoffe in den Lebensmitteln. Wir haben diese Problematik in unserem Institut eingehend studiert und einige Verfahren der chromatographischen Trennung dieser Stoffe mittels DC erarbeitet.Zur Trennung der fettlöslichen Farbstoffe verwendeten wir einerseits DC-Platten mit Aluminiumoxid, andererseits führten wir die Trennung dieser Stoffe auf einer dünnen Stärkeschicht mit umgekehrter Phase durch. Im ersten Falle wurde eine gute Trennung mit Laufmitteln erreicht, die Petroläther und Tetrachlormethan und deren Gemische enthalten. Im zweiten Falle wurde als stationäre Phase Paraffinöl, als mobile Phase eine Gemisch von Methanol, Wasser und Essigsäure im Verhältnis 16 : 3 : 1 und andere Gemische in gleichen Verhältnissen verwendet, die sich bei der Papier-Verteilungschromatographie bewährt haben.Wasserlösliche Farbstoffe wurden auf einer dünnen Schicht von Polyamidpulver getrennt, das auch zu deren quantitativer Isolation aus den Lebensmitteln verwendet wurde. Die besten Ergebnisse wurden mit alkalischen Laufmitteln erzielt, z.B. mit dem Gemisch von Ammoniak, Methanol und Wasser im Verhältnis 5 : 15 : 80.Die erarbeiteten analytischen Verfahren für Nachweis und Trennung fett- und wasserlöslicher Lebensmittelfarben bewährten sich für alle Lebensmitteltypen.

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Résumé L'utilisation des colorants alimentaires solubles dans l'eau et dans la graisse exige des preuves sûres et l'identification exacte de ces substances dans les aliments. Nous avons étudié ces problèmes en détail à notre institut et élaboré quelques procédés de séparation chromatographique de ces substances par chromatographie sur couche mince.A la séparation des colorants solubles dans la graisse, nous avons utilisé d'une part des couches versées sur l'alumine, d'autre part la séparation de ces substances sur une couche mince de fécule à phase inverse. Au premier cas une bonne séparation fut atteinte aux systèmes comprenant l'éther de pétrole et le tétrachlore carbonique, ou leurs mélanges. A l'autre cas on applique, comme une phase ancrée, de l'huile paraffinique, comme une phase mobile, du mélange de méthanol, d'eau et d'acide acétique en proportion 16 : 3 : 1 et d'autres en mêmes proportions qui avaient fait leurs preuves à la chromatographie de séparation sur le papier.Les colorants solubles dans l'eau furent séparés sur une couche mince de poudre polyamidé qui avait été de même appliqué à leur isolement quantitatif de l'aliment. Les meilleurs résultats furent atteints avec les systèmes alcalins p.e. avec le mélange de méthanol, d'ammoniac et d'eau en proportions 5 : 15 : 80.Les procédés analytiques élaborés pour des preuves et les séparations des colorants solubles dans la graisse et dans l'eau prouvèrent leur attestation avec tous les types de matières alimentaires.