Schmallenberg virus epidemic: Impact on milk production,reproductive performance and mortality in dairy cattle in the Netherlands and Kleve district,Germany

Schmallenberg virus (SBV), a novel orthobunyavirus that rapidly spread throughout north-western Europe in 2011, caused congenital malformations in lambs and goat kids (Van den Brom et al., 2012) and newborn calves (Hoffmann et al., 2012). The impact of the SBV epidemic seemed limited however, in terms of the number of affected herds with malformed offspring (European Food Safety Authority, 2012b). Nevertheless, little is known with regard to the overall within-herd impact of SBV infection. The objective of the current study was to quantify the impact of the 2011 SBV epidemic on the productivity of dairy cattle in the Netherlands and the district of Kleve, Germany.     

Measurement of protein turnover in skeletal muscle strips

Isolated porcine and bovine muscle strips were incubated in Krebs Ringer bicarbonate buffer to determine in vitro protein synthesis (PS) and protein degradation (PD) rates to validate the in vitro system for use with livestock species. The addition of 5X plasma concentrations of amino acids to the medium stimulated PS 30%. Addition of 3.5 mM leucine to a leucine-deficient buffer supplemented with amino acids decreased PD 37% and stimulated PS 24%. The addition of .1 U/ml insulin reduced PD 28% and increased PS 30%. Protein degradation was elevated in longitudinally split rat soleus and extensor digitorum longus muscles compared to their contralateral intact muscles. Muscle strips must be removed within 15 min of exsanguination because PD rates become greatly elevated thereafter. ATP concentrations declined during incubation, but the addition of ATP or creatine had no effect on either PD or PS. Neither PD nor PS was affected by the addition of transferrin, fetuin, ascorbate, dexamethasone or indomethacin to the incubation medium. However, muscle strips were sensitive to the addition of triiodothyronine (T3), PD was increased up to 75% as T3 concentration was increased, and PS rates doubled compared to controls. Serum from mature barrows or gilts had no effect on protein turnover, but the addition of 10% and 15% serum from boars increased both PD and PS. With fasted pigs a continual decline in PS occurred over 5 d, whereas PD was elevated at 3 d and then declined to rates comparable to the fed state after 5 d. These data suggest that the in vitro system has application for assessing relative changes that occur in vivo following nutritional, physiological and endocrinological manipulation.     

Epidemiological study of bovine respiratory syncytial virus infections in calves: influence of maternal antibodies on the outcome of disease

A prospective epidemiological survey on bovine respiratory syncytial virus (BRSV) infections in calves was carried out on 21 dairy farms during one BRSV epidemic season. Special attention was paid to the role of maternal antibodies. On 15 farms the spread of the virus was demonstrated during the investigation period and on eight farms this was accompanied by an outbreak of acute respiratory disease. Disease seldom occurred in calves younger than two weeks old and the most severe disease was observed in calves from one to three months old. Although maternal antibodies did not effectively prevent the disease, both the incidence and severity of disease were inversely related to the level of specific maternal antibodies. Two serodiagnostic techniques were compared. In calves older than three months from herds with disease outbreaks associated with bovine respiratory syncytial virus the diagnosis was established in 80 per cent of the animals by an increase in IgG titre against BRSV and in 77 per cent by the detection of BRSV specific IgM. In comparison, only 10 per cent of the calves younger than three months were positive by IgG serodiagnosis, and 51 per cent by IgM serodiagnosis. On farms where the spread of the virus was accompanied by an outbreak of clinical disease more calves were present, a higher proportion of the calves was younger than three months, and calves of all ages were more often housed together.     

Diagnostic assays applied in BVDV control in The Netherlands

In The Netherlands, a voluntary bovine virus diarrhoea virus (BVDV)-free certification programme was started in 1997. After an intake procedure in which all cattle are tested for the presence of BVD virus, a herd obtains the status "BVD-virus-free". To maintain this status a monitoring procedure is executed to verify absence of BVDV circulation in the herd. Several diagnostic tests are used: RT-PCR in bulk milk and pooled blood samples, antigen-ELISA (Ag-ELISA) and antibody ELISA in individual blood samples. Sensitivity and specificity of these tests are discussed. In addition, a diagnostic quick scan has been introduced, consisting of a combination of bulk milk tests for virus and antibody, and antibody tests in samples from young stock. Preliminary results are presented.     

Molecular identification of arbuscular mycorrhizal fungal spores associated to the rhizosphere of Retama raetam in Tunisia

ABSTRACT

Arbuscular mycorrhizal fungi (AMF) are found in the soil of most ecosystems where they form mutualistic associations that affect plants growth. We have investigated the community structure of AMF associated to Retama raetam growing in five regions of Tunisia. The total number of spores was significantly different across sites, ranging from 633 to 1062 spores per 100 g dry soil. A dominance of small spores was revealed. The large subunit region of the rDNA of AMF spores associated to the rhizosphere of R. raetam was sequenced. Sequences clustered into 13 operational taxonomic units. Phylogenetic analysis revealed that the majority of sequences were grouped within Glomeraceae and Claroideoglomeraceae families. Only two sequences were affiliated to the Scutellospora genus. These results suggest the dominance of the genus Glomus in the soil rhizosphere of R. raetam. A correlation between phylogenetic analysis, soil chemicals properties, and AMF community richness was also detected.     

Molecular polymorphism of cytoplasmic DNA in Ficus carica L.: Insights from non-coding regions of chloroplast DNA

The trnL (UAA) intron and the intergenic spacer between the 3′ exon of trnL (UAA) and trnF (GAA) sequences were used as genetic markers for differentiating Ficus carica cultivars and establishing refined genetic relationships. The study was based on 20 fig cultivars, collected from south and centre of Tunisia. Since, the intron was thought to be more variable among close relatives than is the chloroplast spacer. The size of these non-coding regions varied from 554 to 589 and from 989 to 1022 bases pairs for the intron and the combined sequences correspondingly. The average of GC content was 33.9% and 34.6% in the intron and the combined intron and spacer respectively. High values of A + T contents were detected in both data sets and may explain the high proportions of transversions founded. The observed variation pattern of plastid DNA provides evidence of an important genetic diversity. The overall transition/transversion bias (R) was 0.202 in the intron and 0.27 in the combined regions. The RI index of 0.592 indicates that these combined sequences have clearly more homoplasy then the intron (RI = 0.705) and spacer (RI = 0.777) sequences separately. Phylogenetic trees were generated based on maximum parsimony (MP) and neighbor-joining (NJ) analysis of the chloroplast sequences data. Results proved that a typically continuous genetic diversity characterizes the local fig germplasm. In fact, relationships inferred from the cpDNA analysis suggest several clades, which do not show geographical or tree sex correspondence. Although the level of apparent diversity is considerable, we may conclude that non-coding regions of chloroplast genome provide a new and practical opportunity to evaluate genetic diversity and to discriminate fig cultivars. Revealed cytoplasmic DNA markers are reliable to elaborate a molecular data base to conduct management and breeding programs on local fig germplasm.     

The Results of Using Faecal Culture as Confirmation Test of Paratuberculosis‐seropositive Dairy Cattle

A total of 15 822 cattle aged 3 years and older, belonging to 378 randomly selected herds, were tested for paratuberculosis using an absorbed enzyme‐linked immunosorbent assay (ELISA); 3.3% tested positive. This percentage was lowest for the group of cattle aged 3–4 years (2.3%) and highest for cattle with the age of 5–6 years (4.5%). The mean Sample to Positive (S/P) ratio of seropositive cattle vaccinated against paratuberculosis was higher (0.75 ± 0.33) than that of seropositive, non‐vaccinated cattle (0.58 ± 0.26). Faecal samples of 422 ELISA‐positive cattle were cultured for the presence of Mycobacterium avium subsp. paratuberculosis, 12% of these were contaminated. The percentage of non‐contaminated samples with positive culture results was 17.3%, with a substantial difference between vaccinated (1.7%) and non‐vaccinated cattle (20.2%). Of the positive cultures, the number of colonies varied from 1–10 (22% of cultures), 11–100 (22%), to more than 100 (55%). The percentage of ELISA‐positive, non‐vaccinated cattle tested culture‐positive was positively correlated with the magnitude of the S/P ratio. This percentage varied from 12% (S/P ratio 0.3–0.5) to 58% (S/P ratio > 1.1), a result that might have implications for interpretation of the test. In this study, the percentage of ELISA‐positive cattle with positive faecal culture results was limited and these individuals were mostly moderate to heavy shedders.     

Neural mechanisms of foraging

Behavioral economic studies involving limited numbers of choices have provided key insights into neural decision-making mechanisms. By contrast, animals' foraging choices arise in the context of sequences of encounters with prey or food. On each encounter, the animal chooses whether to engage or, if the environment is sufficiently rich, to search elsewhere. The cost of foraging is also critical. We demonstrate that humans can alternate between two modes of choice, comparative decision-making and foraging, depending on distinct neural mechanisms in ventromedial prefrontal cortex (vmPFC) and anterior cingulate cortex (ACC) using distinct reference frames; in ACC, choice variables are represented in invariant reference to foraging or searching for alternatives. Whereas vmPFC encodes values of specific well-defined options, ACC encodes the average value of the foraging environment and cost of foraging.     

MM3-ELISA evaluation of coproantigen release and serum antibody production in sheep experimentally infected with Fasciola hepatica and F. gigantica

During an experimental infection of sheep with Fasciola hepatica or F. gigantica, MM3-SERO and MM3-COPRO ELISA tests were applied to compare the kinetics of antibody production and coproantigen release between the 2nd and 32nd week post-infection (wpi). The Kato-Katz technique was used to measure the kinetics of egg shedding by both Fasciola species (eggs per gram of feces, epg). The kinetics of IgG antibodies for all sheep infected with F. hepatica and F. gigantica followed a similar pattern. Optical density (OD) increased rapidly between the 4th until the 12th wpi, when the highest values were reached and then decreased slowly until the 32nd wpi. Coproantigen levels increased above the cut-off value between 6 and 9 wpi in the F. hepatica group, and between 9 and 11 wpi in the F. gigantica group. The comparison between coproantigen levels and epg indicated that F. hepatica-infected sheep had detectable amounts of coproantigens 4–7 weeks before patency (egg shedding), while F. gigantica-infected sheep had detectable amounts of coproantigens 3–6 weeks before patency. When comparing the kinetics of coproantigen release vs the kinetics of epg, a similar pattern emerged, but with a two-week time-lag in epg, for both F. hepatica and F. gigantica infections. The amount of coproantigen release by each adult was not burden dependent for F. hepatica infection (burden of 33–66 adults), while it was for F. gigantica infection (burden of 17–69 adults). The results demonstrate the usefulness of the MM3-SERO and MM3-COPRO ELISAs as tools for the diagnosis of early as well as long-term fascioliasis infections, and suggest that they can potentially be applied to human fascioliasis even in countries where F. hepatica and F. gigantica co-exist. These tests can be employed not only in the diagnosis, but also in studies on epidemiology as well as pathogenesis and treatment in animals and humans since they allow post-treatment infection monitoring.