Expression of the major outer membrane protein (MOMP) of Chlamydophila abortus, Chlamydophila pecorum, and Chlamydia suis in Escherichia coli using an arabinose-inducible plasmid vector

The ompA genes encoding the 40 kDa major outer membrane protein (MOMP) of Chlamydophila (Ch.) abortus, Ch. pecorum, and Chlamydia (C.) suis were cloned into the arabinose-inducible plasmid vector pBADMycHis, and recombinant MOMPs (rMOMP) from the three chlamydial species were expressed at high levels in Escherichia (E.) coli. The proteins lacking the 22 aa N-terminal signal peptide were expressed as insoluble cytoplasmic inclusion bodies which were readily purified using immobilized metal-affinity chromatography. The rMOMPs including the N-terminal signal peptide were expressed and translocated as a surface-exposed immunoaccessible protein into the outer membrane of E. coli. Transformants expressing this full-length rMOMP were significantly reduced in viability. Purified native elementary bodies (EB) and rMOMPs of the three chlamydial species purified from the E. coli cytoplasm were used for immunization of rabbits. The resulting sera were analysed for their ability to recognize homologous and heterologous rMOMP and native EB. When testing rMOMP antisera against rMOMP and EB antigens, marked cross-reactivities were detected between the three species. Using EB antisera and rMOMPs as antigens, a significant species-specific reactivity was measured.     

Combining isotopic signatures of n(Sr)/n(Sr) and light stable elements (C,N, O,S) with multi-elemental profiling for the authentication of provenance of European cereal samples

The aim of this work (from the FP6 project TRACE) was to develop methods based on the use of geochemical markers for the authentication of the geographical origin of cereal samples in Europe (cf. EC regulations 2081/92 and 1898/06). For the first time, the potential usefulness of combining n(⁸⁷Sr)/n(⁸⁶Sr) and δ¹³C, δ¹⁵N, δ¹⁸O and δ³⁴S isotopic signatures, alone or with key element concentrations ([Na], [K], [Ca], [Cu] and [Rb], progressively identified out of 31 sets of results), was investigated through multiple step multivariate statistics for more than 500 cereal samples collected over 2 years from 17 sampling sites across Europe representing an extensive range of geographical and environmental characteristics.     

Wheat blast caused by Magnaporthe oryzae pathovar Triticum is efficiently controlled by the plant defence inducer isotianil

Journal of Plant Diseases and Protection - Wheat blast caused by Magnaporthe oryzae pv. Triticum is an upcoming threat to wheat cultivation worldwide. The disease crossing over to wheat first...     

Real-time RT-PCR detection of Citrus bark cracking viroid (CBCVd) in hops including an mRNA-based internal positive control

Journal of Plant Diseases and Protection - Citrus bark cracking viroid (CBCVd), formerly known as pathogen in the genus Citrus and first detected in Slovenian hops in 2014, threatens hop production...     

Prevalence of Chlamydia infections in breeding sows and their importance in reproductive failure

To investigate the prevalence of chlamydial infection and their significance for reproductive disorders in sow breeding herds in Germany, blood samples of 1493 pigs were taken for a serological survey by enzyme-linked-immunosorbent-assay (ELISA). Antibodies to Chlamydiae were found in 33% of the samples, in all herds investigated responders could be detected. The rate of seropositive animals in different farms varied from 4.3% to 72.7%. The percentage of positive responders in the farms correlated positively with the occurrence of MMA-syndrome (mastitis, metritis, agalactia), return to oestrus and diseases of the piglets. Also these herds showed less weaned piglets per sow and litter. Comparison of seronegative and seropositive sows within single farms revealed also worse reproductive data for seropositive sows. A significant relationship could be found between farms with a high quota of sero-positive sows and poor hygiene status as well as poultry keeping. As a second step 124 cervical swabs and 9 aborted piglets were investigated for chlamydial antigen by capture-ELISA and polymerase chain reaction (PCR). Using the capture-ELISA for investigation only 3 probes with chlamydial antigen could be detected, however, examination by the more sensible PCR gave positive results in 50% of the probes. 20% of the PCR-positive animals were clinically healthy sows, 80% of the PCR-positive probes originated from sows with reproductive disorders. A significant relationship could be shown between PCR-positive probes and the incidence of abortion and litters with stillborn piglets and piglets with low viability. Swabs from 93 of the 124 sows were investigated as well for other bacterial pathogens of reproductive disorders. A high degree of micro-organisms of different species could be detected in 70% of the samples of sows with reproductive disorders and in 35% of the samples of clinically healthy sows. Species differentiation of the chlamydial antigen positive samples was done by southern blot hybridisation. Herewith C. psittaci could be diagnosed in all positive samples. Additionally 8 probes revealed a mixed infection with C. psittaci and C. trachomatis. The results of the present study show, that the prevalence of chlamydial infections in breeding herds is high and underline the importance of chlamydial infections for reproductive disorders. Single chlamydial infections as well as mixed infections with other pathogens must be considered.     

Carotenoids in diets for salmonids: I. Pigmentation of rainbow trout with the individual optical isomers of astaxanthin in comparison with canthaxanthin

Visual and chemical determinations revealed that the individual optical isomers of astaxanthin were more efficacious than canthaxanthin in pigmenting the flesh of rainbow trout (Salmo gairdneri, Richardson). The visual assessment of the astaxanthin pigmentation by means of a colour scale correlated well with the chemical determination. The same utilization was found for all three astaxanthin isomers, (3S, 3′S)-, (3R, 3′S)-, (3R, 3′R)-astaxanthin and a 1 : 2 : 1 mixture of the three isomers.No epimerization took place at the chiral centres of C-3 and C-3′ in astaxanthin. Astaxanthin and canthaxanthin were not interconverted. No metabolites were detected in the flesh.     

Thymomas in rabbits: clinical evaluation, diagnosis, and treatment

Thymomas are rarely recorded in rabbits, and the literature includes comparatively few cases. Medical records were reviewed to identify all pet rabbits in which a mediastinal mass was diagnosed between Feb 2007 and Jan 2010. Signalment, history, clinical signs, diagnostic work-up (including laboratory data, diagnostic imaging, and ultrasound-guided fine-needle aspiration of the mediastinal mass), treatment modalities, survival time, and histologic findings were evaluated. Cytologic and/or histopathologic examinations revealed thymomas in all rabbits with mediastinal masses (n=13). Rabbits with thymomas showed clinical signs of dyspnea (76.9%), exercise intolerance (53.9%), and bilateral exophthalmos (46.2%). In seven rabbits the thymoma was removed surgically. Two rabbits were treated conservatively, and four rabbits were euthanized because of their poor clinical condition. The two rabbits that underwent surgery were euthanized 6 mo and 34 mo later. Mediastinal masses in rabbits appear to be more common than previously believed and consist primarily of thymomas rather than thymic lymphomas. Cytology of samples collected by ultrasound-guided fine-needle aspiration is an accurate diagnostic tool for the identification of thymomas in rabbits. Due to a high rate of perioperative mortality, intensive perioperative care and the provision of a low-stress environment are recommended for a successful thoracotomy.     

Biochar application to sandy soil: effects of different biochars and N fertilization on crop yields in a 3-year field experiment

During the past years, most biochar studies were carried out on tropical soils whereas perennial field experiments on temperate soils are rare. This study presents a 3-year field experiment regarding the effects of differently produced biochars (pyrolyzed wood, pyrolyzed maize silage, hydrothermal carbonized maize silage) in interaction with digestate incorporation and mineral N fertilizer application on soil C and N, crop yields of winter wheat, winter rye and maize and the quality of winter wheat. Soil C and plant available potassium were found to be significantly positive affected by pyrolyzed wood biochar whereas the latter only in combination with N fertilization. Crop yields of winter wheat, winter rye and maize were not affected by biochar and showed no interaction effects with N fertilizer supply. Wheat grain quality and nutrition contents were significantly affected by biochar application, for example, highest amounts of phosphorus, potassium and magnesium were determined in treatments amended with pyrolyzed maize silage biochar. Biochar induced an improved availability of plant nutrients, which apparently were not yield limiting in our case. These results limit the potentials of biochar for sustainable intensification in agriculture by increasing crop yields for the temperate zones. However, detection of other environmental benefits requires further investigations.     

Seroprevalence of anaplasmosis among cattle in Switzerland in 1998 and 2003: no evidence of an emerging disease

Anaplasma marginale infection in Europe has been limited to the Mediterranean and eastern countries, to Austria and to very sporadic cases in Switzerland. There are no reports of its occurrence in the countries north of Switzerland. A severe outbreak of anaplasmosis in August 2002 in a cattle farm in the canton Grisons, Switzerland, north of the Alps, with more than 300 cattle that had to be culled, came unexpected and gave reason to hypothesize presence of an increased yet undetected prevalence of A. marginale in Switzerland. Randomly selected bovine serum samples collected in 1998 and 2003 were tested using a competitive inhibitory ELISA (cELISA) to test the hypothesis. Our validation of the diagnostic sensitivity and specificity of this test, done in the outbreak herd, yielded 99.2 and 83.3%, respectively, probably underestimating the true specificity. The true seroprevalence of anaplasmosis in Swiss cattle determined by cELISA was likely to be zero with upper 95% confidence limits of 2.49% in the canton Grisons and 1.17% in the rest of Switzerland, respectively, in 1998. For 2003, these estimates were even lower. There was no significant difference in apparent prevalences between 1998 and 2003. In search of a possible reservoir, three chamoises out of 46 free ranging wild ruminants from the Swiss National Park, Grisons, tested positive in the cELISA. This reaction is in accordance with A. marginale or a cross reacting agent such as Anaplasma ovis. From our results we conclude that the hypothesis of an increased prevalence of anaplasmosis in cattle in Switzerland must be rejected.