Acceptance of alternative disease control strategies in the European Union

The European Commission intends to change its animal disease control strategy following the slogan "Prevention is better than cure". Vaccination and diagnostics should play a major role in emergency situations. A policy paper regarding the use of vaccines has been discussed with all Member States and the main aspects were evaluated in a questionnaire. In principle, the majority of Member States are in favour of a future strategy in which vaccination is replacing culling. However, questions regarding the pathogen freedom and the trade of vaccinated animals and animal products from vaccinated animals still remain open.     

Screening for in planta protein-protein interactions combining bimolecular fluorescence complementation with flow cytometry

ABSTRACT: Understanding protein and gene function requires identifying interaction partners usingbiochemical, molecular or genetic tools. In plants, searching for novel protein-proteininteractions is limited to protein purification assays, heterologous in vivo systems such as the yeast-two-hybrid or mutant screens. Ideally one would be able to search for novel proteinpartners in living plant cells. We demonstrate that it is possible to screen for novel proteinproteininteractions from a random library in protoplasted Arabidopsis plant cells and recoversome of the interacting partners. Our screen is based on capturing the bi-molecularcomplementation of mYFP between an YN-bait fusion partner and a completely random preyYC-cDNA library with FACS. The candidate interactions were confirmed using in plantaBiFC assays and in planta FRET-FLIM assays. From this work, we show that the wellcharacterized protein Calcium Dependent Protein Kinase 3 (CPK3) interacts with APX3,HMGB5, ORP2A and a ricin B-related lectin domain containing protein At2g39050. This isone of the first random in planta screens to be successfully employed.     

Mammary tumor recurrence in bitches after regional mastectomy

Objectives— To investigate the histologic diagnosis and incidence of new mammary tumor growth in the remaining mammary chain tissue after regional mastectomy.
Study Design— Prospective clinical study.
Animals— Female dogs (n=99) that had excision of a single mammary tumor.
Methods— Female dogs that had regional mastectomy to remove a single tumor were followed for ≥1 year postoperatively. Data regarding tumor type, tumor recurrence, and development of metastasis were recorded.
Results— Fifty-seven (58%) dogs developed a new tumor in the ipsilateral mammary chain after the 1st surgery; 77% had repeat surgery. There was no significant correlation between the time to new tumor development and the histologic diagnosis for the 1st and 2nd tumor types. In 31 dogs, the histologic diagnosis for initial and subsequent tumors was identical and there was a significant correlation such that dogs with an initial malignant tumor are likely to develop another malignant tumor ( P =.0089). The histologic classification of the new tumor was likely to be malignant if it was located close to the side where the initial tumor had been removed ( P =.026).
Conclusions— Our results show that 58% of dogs developed a new tumor in the remaining mammary glands of the ipsilateral chain after regional mastectomy for removal of a single tumor.
Clinical Relevance— This should be taken into account when deciding on the surgical management (radical or regional mastectomy) in dogs with single mammary tumors.     

Evaluation of real-time RT-PCR assay for the routine intra vitam diagnosis of classical swine fever

The aim of the study was to evaluate the diagnostic quality of the real-time RT-PCR assay described by Hoffmann et al. [Hoffmann, B., Beer, M., Schelp, C., Schirrmeier, H., Depner, K., 2005. Validation of a real-time RT-PCR assay for sensitive and specific detection of classical swine fever. J. Virol. Methods 130, 36-44] for the routine intra vitam diagnosis of classical swine fever (CSF). We compared the assay with conventional diagnostic methods by using defined diagnostic material from an animal experiment with pigs showing different clinical forms of CSF. Compared to virus isolation and antigen ELISA an enhanced sensitivity of the real-time RT-PCR could be shown. We were able to detect all infected pigs regardless of the clinical course of CSF. CSF infection was detected already during the incubation period, during the entire clinical phase as well as at the beginning of convalescence when the first antibodies were detected and no virus could be isolated any more. In most cases, positive PCR results were obtained 2 days earlier than with virus isolation and 2-4 days earlier than with the antigen ELISA.     

Untersuchungen zum Voltinismus der Rotgelben Kiefern-BuschhornblattwespeNeodiprion sertifer (Geoffroy) in Südtirol und Trentino (Hym., Diprionidae)

During an outbreak ofN. sertifer in Brentonico (on Monte Baldo, Trentino, North Italy), in altitudes of 1,400–1,600 m, surveys were made in 1996 and 1997 on the voltinism of these short-day-Diprionide, wherely over 2,000 cocoons were reared (fig. 1; tab. 1–4). Therefore L4–5 larvae ofN. sertifer from different altitudes were transferred to Denno (430 m) and to Brixen (550 m) and were reared there under outdoor conditions. These surveys were completed with further observations of populations from South Tyrol, of St. Ulrich Val Gardena (1,200 m) and italian provenances from Riolo Terme (150 m) in Brixen (fig. 2). These comparative studies revealed a correlative connection between the time of cocoonspinning and adult emergence. They showed the following principle: the later the cocoonspinning, the earlier the adult emergence. Early spinners (e. g., May/June) emerge only late in October, while late spinners (e. g., early August) emerge as soon at the end of August. In the case of late cocoon spinning the obligatory summer cocoon diapause is so reducted, eventually—at the start of the short-day phase (with only 15 hours brightness), in early August—leading to a subitan development. These different cocoon spinning periods cause varying flywaves, W1–W4, ofN. sertifer, similarly to other Diprionidae; this seems evident also by pheromon-trap catches (fig. 2: 3). The possible variable combinations of these fly-waves are discussed. It is shown that voltinism and diapause behaviour ofN. sertifer are essentially conditioned by the photoperiod and correlated climatic factors. The determined low diapause rate, of just 1–2%, is connected with the southern and warmer location of the outbreak area of Brentonico (45° 45’ N). The exceptional longevity of adults, which can amount to several weeks, also plays an important role.     

Liver histology and histomorphometry in hybrid sorubim (Pseudoplatystoma reticulatum × Pseudoplatystoma corruscans) reared on intensive fish farming

This study aimed to characterize the liver histology and histomorphometry in sorubim hybrid of different categories (nursery, growth and grow‐out) reared on fish farming. The categories were defined considering body weight (BW): nursery category (n = 5): BW = 37.06 ± 6.00 g (31.6–45.3 g); growth category (n = 5): BW = 310.40 ± 53.80 g (242.1–376.4 g) and grow‐out category (n = 5): BW = 874.28 ± 27.59 g (846.2–913.1 g). Liver fragments were processed to paraffin inclusion, and sections were stained by haematoxylin and eosin (H&E), PAS (Periodic Acid Schiff) and Perl's staining to histology, histomorphometry and density volumetric of liver structures; glycogen analysis and to detect ferric irons (Fe³⁺) respectively. The hepatosomatic index decreased between the categories (P < 0.01). The percentage of PAS‐positive hepatocytes in the nursery category was higher (P < 0.05) in relation to the growth and grow‐out categories. The hepatocytes from all fish were positive to Perl's staining. The density volumetric of liver structures did not differ among categories except to blood vessels were higher (P < 0.01) in the nursery and growth. The area (μm²) and perimeter (μm) of hepatocytes, and the area (μm²), perimeter (μm) and volume (μm³) of the nuclei from grow‐out fish were lower (P < 0.01) than those from the nursery and growth categories. Changes in morphometric characteristics of hepatocytes may result from metabolic changes associated with body growth surubins; therefore, these morphometric characteristics of liver tissue can be used as functional biomarkers for the assessment of fish health and nutrition status.     

Segregation of microsatellite loci in second generation hybrid striped bass (<Emphasis Type="Italic">Morone saxatilis</Emphasis> × <Emphasis Type="Italic">Morone chrysops</Emphasis>)

Within the framework of a larger project aimed to assess the potential of second generation hybrid striped bass for German aquaculture the genotypic segregation of five microsatellite loci was analysed in two progeny lots (n = 74 and 76, respectively). There was no consistent correlation between microsatellite genotypes and phenotypic category (white bass, hybrid, or striped bass). None of the individuals expressed neither only white bass nor only striped bass genotypes at all five loci. On the other hand, only hybrid genotypes at all five loci were detected in three individuals of lot 1 and four individuals of lot 2. Single loci tests for conformity of microsatellite genotypic segregation with Mendelian rules revealed significant deviations (P < 0.05) in four cases for lot 1 and in three cases for lot 2. If pooled over all five loci, both lots displayed highly significant deviations (P < 0.01) with an excess of hybrid genotypes and a deficiency of white bass genotypes. It is concluded that stabilizing selection performed on hybrid genotypes might be a suitable approach for practical aquaculture in Europe if the goal is to become independent of first generation hybrid fry supply and/or if establishing domesticated brood stocks of both parental species is impossible. However, more detailed studies on the characteristics and performance of multiple hybrid generations are needed.