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EN
Coronavirus 800 (18) propagated in the lung cells of the calf foetus (16) was purified and concentrated by: a) differential centrifugation, b) precipitation with PEG-6000, c) repeated haemadsorption-elution assay (9). The samples were examined under a transmissive microscope after prior negative staining. There were found many virions (Fig 1 and 2) with a typical morphology of coronavirus (2, 12, 13, 14, 15, 17). Besides, there were observed some particles with internal nucleoprotein in the form of a spisal or S-shape, and empty particles (Fig 5) with bilateral concave surfaces (fig 6). Unusual peplomers forming short (Fig 7, 8) and long filaments (Fig 9, 10) were also seen. However, to identify these unusual structures as corona- viruses or coronavirus substructures (Fig 11, 12) it is necessary to employ a specific method using labelled antibodies.
EN
The nested PCR technique (Borchers and Slater, 1993) was applied for the diagnosis of EHV-1 infections. DNA samples were isolated from livers of miscarried fetuses or dead foals. After amplification we detected EHV-1 specific sequences in 9 out of 15 fetuses and in 2 out of 6 foals. PCR results were compared to results of routinely performed immunofluorescent detection of viral antigen in cryosections of tissues. Six tested fetuses were FAT-positive, 7 FAT-negative and 2 were questionable. One FAT-negative fetus and two questionable were PCR-positive. All foals were FAT-negative. We have demonstrated that PCR is more sensitive than FAT and allows verification of inconclusive results of antigen detection. PCR is also suitable for detection of EHV-1 specific sequences in archival samples frozen up to 3 years.
EN
110 blood samples from clinically healthy mares of English breed, half-blood and the Wielkopolska race were tested for the presence of EHV-1 and EHV-2 using nested PCR. 15 samples were EHV-1-positive, 24 samples were EHV-2-positive, whereas only 4 samples were both EHV-1 and EHV-2 positive. The virus was isolated from PBLs in equine dermal cell cultures by co-cultivation or by culture inoculation with cell lysates derived from PBLs. A total of 14 strains were isolated from EHV-2 and EHV-1/EHV-2-positive samples. However, all of them were identified by nPCR as being type 2. Since no EHV-1 was isolated, even from dually infected leukocytes, it was concluded that the presence of EHV-2 does not stimulate in-vitro isolation of EHV-1 from infected leukocytes. It is tempting to speculate that such stimulatory effects in-vivo may involve EHV-2-induced immunosupression. Despite any possible mechanism of EHV-1 stimulation, it seems that EHV-2 does not play a significant role in the epidemiology of EHV-1-caused miscarriages in horses since mixed infections are rather rare.
EN
Equine herpesvirus type 1 (EHV-1) is one of the major horse diseases, causing considerable worldwide losses. A variety of techniques, including nested PCR, have been used to diagnose EHV-1 infections. In this paper, a real-time PCR assay that uses non-specific SYBR Green I® fluorochrome for the detection of EHV-1 DNA is described. This method does not require post-amplification manipulations, thereby reducing the risk of cross-contamination. The assay was sensitive enough to detect EHV-1 sequences in neuronal cell cultures and also different clinical samples. The technique is specific: it was not reactive with other herpesviruses or opportunistic bacterial pathogens such as Escherichia coli, Staphylococcus epidermidis and Enterococcus faecium. In comparison to virus isolation or the nested PCR used previously, the test was more sensitive and should be useful for the common diagnosis based on its specificity and rapidity.
EN
Producing high quality offspring of good physiological performance, able to survive to independence and, then, to reproductive maturity is a major component of life history strategies. The ability of nestling altricial birds to develop a good physiological condition depends to a large extent on the amount and quality of food provided by parents, as well as other aspects of parental care. We hypothesized that experimental changes to the original brood size should affect both parental Blue Tits and their offspring, resulting in corresponding changes in the body condition of the nestlings. Over two breeding seasons, using two habitat sites, we conducted an experiment with two manipulative treatments applied to broods of three-day-old nestlings — the reduction or enlargement of broods by three nestlings, and one non-manipulative control treatment. Our aim was to test whether the experiment would affect a number of different measures of nestling condition: blood concentrations of hemoglobin and glucose, heterophil-to-lymphocyte ratio and morphometric condition index, all being analyzed when the nestlings were 13 days old. We found no effect in the case of hemoglobin, despite the fact that it had previously been shown to be sensitive to large-scale differences in trophic conditions between habitats and years and to the experimental removal of nest parasites. All the remaining variables, i.e. heterophil-to-lymphocyte ratio, glucose concentration and morphometric condition index, responded to the experimental treatments, showing different but reasonable patterns of variation. We suggest that an experimental increase in brood size definitely hinders the development of nestling physiological condition, but even an experimental reduction of broods can affect some physiological indicators (glucose), probably because of readjustments in the feeding rate.
PL
Eksperyment przeprowadzono w latach 2005-2006 w dwóch środowiskach, parkowo-ogrodowym i leśnym, w ramach długoterminowego projektu badań nad sikorami w Łodzi i okolicy. W obu środowiskach zlokalizowane są powierzchnie badawcze zaopatrzone w drewniane skrzynki lęgowe, zasiedlane przez dziuplaki wtórne. Na lęgach modraszki przeprowadzono eksperyment polegający na zmianie wielkości lęgów o 3 pisklęta w trzecim dniu po wykluciu. Znajdujące się w tym samym stadium lęgi (n = 135) losowo przypisywano do zabiegów zmniejszania (n = 29) i zwiększania (n = 29) oraz roli kontrolnej (n = 77). Zabieg doświadczalny, rok i środowisko były traktowane jako czynniki, zaś jako zmienne zależne przyjęto następujące charakterystyki: stężenie hemoglobiny, stosunek liczby heterofili do limfocytów, stężenie glukozy we krwi, morfometryczny indeks kondycji (wartości resztowe z regresji masy ciała na długość skrzydła) oraz sukces wylotu (proporcja liczby podlotów w stosunku do zmienionej w wyniku zabiegu eksperymentalnego wielkości lęgu). Zmienne zależne uzyskano w wyniku badania krwi i pomiarów morfometrycznych 13-dniowych piskląt, dla podprób 5 osobników na gniazdo. Dla każdej podpróby wyliczano średnią, która była traktowana jako obserwacja jednostkowa. Nie wykryto wpływu eksperymentu na średnie stężenie hemoglobiny Wykryto istotny wpływ na stosunek heterofili do limfocytów, stężenie glukozy, morfometryczny indeks kondycji i sukces wylotu. (Tab. 2, Fig. 1). Poza wpływem czynnika eksperymentalnego w niektórych przypadkach istotny okazał się także wpływ roku, środowiska i interakcji czynników. W sumie wyniki tworzą skomplikowany, ale dający się zrozumiale interpretować wzorzec. Poszczególne zmienne zależne reagowały na czynnik eksperymentalny w odmienny sposób, co wydaje się mieć związek z odmienną skalą czasową i fizjologicznym poziomem ich kształtowania się w trakcie życia piskląt.
EN
Long-term annual variation in the timing of egg laying, clutch size and relationship between clutch size and the progress of the season was analysed for the Pied Flycatcher Ficedula hypoleuca nesting in a mature deciduous woodland in central Poland in 2002–2010. The earliest mean egg laying date was 8 May (2005) and the latest 18 May (2008), resulting in the maximum difference of 10 days between averages for years. No long- term trend was found. The total average of annual mean laying dates was 12 May. For all nine years the average of annual mean clutch sizes was 6.54 ± 0.28 (SE) eggs; for individual seasons mean clutch size ranged from 6.0 to 7.1 eggs but differences among years were not significant. Clutch size clearly tended to decline with the progress of the breeding season within years, with some variation between years; correlation for pooled standardized data was –0.49. This supports the idea that in long-distance single-brooded passerine birds clutch size should decrease with the course of the breeding season due to progressively deteriorating food conditions.
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