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.     

Identification of Cytokeratins in Bovine Sperm Outer Dense Fibre Fractions

Outer dense fibres (ODF) are important substructures of mammalian sperm tails that are involved in the regulation of sperm motility. In this study, we investigated the identity of several sodium dodecyl sulphate (SDS)‐insoluble ODF proteins. Bovine ODF were purified by separating sperm heads and tails using ultrasound and Percoll^® density gradient centrifugation. Sperm flagella were treated with the detergent cetyltrimethylammonium bromide (CTAB). CTAB‐insoluble material, which reportedly represents the ODF fraction, was collected, and electron microscopy confirmed a highly purified ODF fraction. We found after solubilization of this fraction with SDS that high amounts of insoluble material were retained after centrifugation. SDS‐insoluble material was collected and quantitatively dissolved in 8 M urea. SDS‐gel electrophoresis in the presence of urea revealed polypeptides with apparent molecular masses of approximately 25, 43, and 50 kDa. Subsequent immunoblotting with anti‐cytokeratin antibodies detected two urea‐soluble, SDS‐insoluble proteins with apparent molecular masses of 45 and 66 kDa. The 45‐kDa protein was identified as cytokeratin 19. An antibody reacting with a palette of cytokeratins (CK 1–18 and CK 20), KL1, was the only antibody that reacted with the 66‐kDa polypeptide. We conclude that sperm ODF fractions contain at least one each of type I and type II intermediate filaments. As keratins and intermediate filaments are described as rope‐like structures, we suggest that these intermediate filaments play an important structural or tension‐bearing role in sperm flagella.     

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.     

Isolation and pathogenicity of Colletotrichum spp. causing anthracnose of strawberry in south west Spain

Anthracnose, caused by Colletotrichum spp., is a major disease of cultivated strawberry, Fragaria × ananassa. This study identifies the Colletotrichum spp. which causes strawberry anthracnose in the southwest of Spain. A survey of the region was carried out, and the strains isolated were identified as C. acutatum by using the polymerase chain reaction (PCR) with genus and species-specific primers, demonstrating that this species is currently the causal agent of strawberry anthracnose in the studied region. The pathogenicity of C. acutatum and C. gloeosporioides strains was evaluated on two principal strawberry cultivars (cvs Camarosa and Ventana) under field conditions, the latter being more pathogenic than the former.     

Natural hybridization between wheat (Triticum aestivum L.) and its wild relatives Aegilops geniculata Roth and Aegilops triuncialis L.

BACKGROUND

Cultivated bread wheat (Triticum aestivum L.) spontaneously hybridizes with wild/weedy related Aegilops populations, but little is known about the actual rates at which this hybridization occurs under field conditions. It is very important to provide reliable empirical data on this phenomenon in order to assess the potential crop–wild introgression, especially in the context of conducting risk assessments for the commercialization of genetically modified (GM) wheat, as gene flow from wheat to Aegilops species could transfer into the wild species genes coding for traits such as resistance to herbicides, insects, diseases or environmental stresses.

RESULTS

The spontaneous hybridization rates between wheat and A. geniculata and A. triuncialis, which are very abundant in the Mediterranean area, have been estimated for the first time in the northern part of the Meseta Central, the great central plateau which includes the largest area of wheat cultivation in Spain. Hybridization rates averaged 0.12% and 0.008% for A. geniculata and A. triuncialis, respectively. Hybrids were found in 26% of A. geniculata and 5% of A. triuncialis populations, at rates that can be ≤3.6% for A. geniculata and 0.24% for A. triuncialis.

CONCLUSION

The detection of Aegilops spp.–wheat hybrids in Aegilops populations indicates that gene flow can occur, although wheat is considered a crop with a low-to-medium risk for transgene escape. These data on field hybridization rates are essential for GM wheat risk assessment purposes. © 2023 Society of Chemical Industry.     

Effects of Pre-Stratification Storage Conditions on Black Walnut Seed Post-Stratification Germination Capacity

The objective of this work was to quantify changes in the germination capacity of Juglans nigra seeds following storage and then artificial stratification. Seeds were stored at 5 and –20 °C under 5% and 14% internal moisture content (IMC) for 0, 8, 10, and 12 months under conditions with oxygen or under vacuum. Germination percentage (GP) was affected by storage temperature, IMC, and storage period. Seeds stored with 5% IMC succeeded in germinating regardless of the temperature. Seeds stored with 14% IMC did not germinate (–20 °C) or germinated during storage (5 °C). The GP was reduced with the time of storage in seeds at –20 °C and 5% IMC and this effect was avoided in storage under vacuum. In conclusion, the germination capacity of black walnut seeds did not change for up to 12 months at low above-zero temperature and low IMC (5 °C, 5% IMC), being the most cost-effective alternative for long-term storage of black walnut seeds in commercial nurseries. Vacuum storage avoided the decrease of around 50% in average germination capacity of seeds stored for a year at –20 °C.     

Seasonal and temporal changes in species use of the landscape: how do they impact the inferences from multi-scale habitat modeling?

Context

Multi-scale approaches to habitat modeling have been shown to provide more accurate understanding and predictions of species-habitat associations. It remains however unexplored how spatial and temporal variations in habitat use may affect multi-scale habitat modeling.

Objectives

We aimed at assessing how seasonal and temporal differences in species habitat use and distribution impact operational scales, variable influence, habitat suitability spatial patterns, and performance of multi-scale models.

Methods

We evaluated the environmental factors driving brown bear habitat relationships in the Cantabrian Range (Spain) based on species presence records (ground observations) for the period 2000–2010, LiDAR data on forest structure, and seasonal estimates of foraging resources. We separately developed multi-scale habitat models for (i) each season (spring, summer, fall and winter) (ii) two sub-periods with different population status: 2000–2004 (with brown bear distribution restricted to the main population nuclei) and 2005–2010 (with expanding bear population and range); and (iii) the entire 2000–2010 period.

Results

Scales of effect remained considerably stable across seasonal and temporal variations, but not the influence of certain environmental variables. The predictive ability of multi-scale models was lower in the seasons or periods in which populations used larger areas and a broader variety of environmental conditions. Seasonal estimates of foraging resources, together with LiDAR data, appeared to improve the performance of multi-scale habitat models.

Conclusions

We highlight that the understanding of multi-scale behavioral responses of species to spatial patterns that continually shift over time may be essential to unravel habitat relationships and produce reliable estimates of species distributions.

     

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.