Detection and quantification of Anaplasma marginale DNA in blood samples of cattle by real-time PCR

A TaqMan-based real-time PCR assay was developed for the diagnosis of Anaplasma marginale infection of cattle. The established assay was proven to be highly specific, since no cross-reactions were observed with other Anaplasma species of ruminants, including the closely related Anaplasma centrale, or other haemoparasites of ruminants (Anaplasma bovis, Anaplasma ovis, Anaplasma phagocytophilum, Babesia bovis, Babesia bigemina, Theileria annulata and Theileria buffeli). The detection limit was equal to that of nested (n)PCR (10(1) copies of standard DNA and 3 x 10(1) infected erythrocytes ml(-1) of blood). The assay was also reproducible, as shown by satisfactory low intra-assay and inter-assay coefficients of variation. Fifty-four blood samples of ruminants (cattle, n = 51; sheep, n = 2; goats, n = 1), that had been tested previously by reverse line blot (RLB) hybridisation, were subjected to an nPCR assay and the newly established real-time PCR assay. By using real-time PCR, A. marginale DNA was detected in 39/51 bovine samples, with DNA titres ranging from 3.60 x 10(3) to 5.70 x 10(8) copies ml(-1) of blood, whereas sheep and goat samples tested negative. The concordance with nPCR was 100%, whereas a unique sample that had tested negative by RLB gave positive results by nPCR and real-time PCR. The established assay could overcome the limitations of existing diagnostic methods, allowing for simultaneous detection and quantification of the A. marginale DNA in bovine blood, that is essential to support the clinical diagnosis, to assess the carrier status of the animals and to evaluate the efficacy of vaccines and antirickettsial drugs.     

The Impact of Atmospheric Deposition of Non-Acidifying Substances on the Quality of European Forest Soils and the North Sea

In the pilot study ESQUAD the impact of atmospheric deposition of three heavy metals (cadmium, copper and lead) and two persistent organic pollutants (benzo(a)-pyrene and lindane) on the quality of European soils and seawater has been calculated. Calculations have been made of atmospheric transport and deposition using a detailed emissions database for Europe. This enabled deposition maps to be produced to a resolution of approximately 50 km. The distribution of pollutant concentrations in forest soils was calculated for each grid cell using a database of soil property parameters in Europe. For the North Sea, a model was used to map long-term concentrations in water and sediment, which are due to atmospheric deposition and other, non-atmospheric sources. The model calculations allowed detailed comparisons of deposition fluxes and concentrations of the substances studied with critical loads and environmental quality threshold values, including critical loads. Although significant uncertainties were identified, the study gives insight in how threshold exceedance rates in Europe relate to pollutant type, threshold type, environmental compartment and chemophysical phase (adsorbed, dissolved). For all pollutants and for all compartments exceedances were calculated for at least some of the quality thresholds that were chosen.     

永嘉早香柚授粉及保水措施试验

为探索永嘉早香柚的低产原因和降低内裂果措施。进行了授粉和保水措施试验，结果表明，通过授粉能有效提高永嘉早香柚的座果率；授粉配合保水措施能有效降低永嘉早香柚的内裂果和裂瓣率。     

Natural variation in fruit characteristics,seed germination and seedling growth of <Emphasis Type="Italic">Adansonia digitata</Emphasis> L. in Benin

Adansonia digitata (baobab tree), a multipurpose tree species, occurs throughout semi-arid and arid zones of Africa. Its survival is, however, threatened by bush fire, over-exploitation, grazing and a lack of natural regeneration. The extent of variation in fruit characteristics, seed germination and seedling traits of the baobab tree in Benin, was evaluated at climatic zone level. 1,200 fruits were sampled in each of the three climatic zones of Benin for morphological assessment and to assess germination rate and seedling growth dynamics according to the climatic zones, the used substrate and the scarification of the seed coat. There were significant differences in fruit characteristics not only between climatic zones but also between individuals from the same zone and within-trees. Using mechanical scarification on freshly-collected baobab seeds negatively affected the germination rate of baobab seeds sampled in the Guinean and Sudano-Guinean zones of Benin. The best-germination rate was recorded for non-treated seeds from the Guinean zone, up to 57% on day 25. All seeds germinated best on the sand substrate, but supplying organic matter promoted further seedling growth after 11 days of germination. Based on these observations we propose some strategies for efficient ex situ conservation of baobab in Benin.     

An experimental sod seeder

The design and development of an efficient sod‐seeder for the Ngongoni Veld of Natal is described.     

Feline vaccine‐associated sarcomagenesis: Is there an inflammation‐independent role for aluminium?

Aluminium has been found in feline vaccine‐associated sarcomas. In this study, we investigated the potential for aluminium to contribute directly to tumourigenesis. Our results indicated that an aluminium hydroxide adjuvant preparation was cytotoxic and mutagenic in human‐Chinese hamster ovary (CHO) hybrid cells in vitro. Moreover, CHO cells deficient in DNA double strand break (DSB), but not single‐strand break (SSB), repair, were particularly sensitive to aluminium exposure compared with repair proficient cells, suggesting that aluminium is associated with DSBs. In contrast to CHO cells, primary feline skin fibroblasts were resistant to the cytotoxic effects of aluminium compounds and exposure to an aluminium chloride salt promoted cell growth and cell cycle progression at concentrations much less than those measured in particular feline rabies vaccines. These findings suggest that aluminium exposure may contribute, theoretically, to both initiation and promotion of tumours in the absence of an inflammatory response.     

Comparison of biopsy samples obtained using standard endoscopic instruments and the harmonic scalpel during laparoscopic and laparoscopic-assisted surgery in normal dogs

Objectives— To compare tissue specimens of canine abdominal organs collected by standard techniques (ST) and harmonic scalpel (HS) and to determine the effect of each technique on wound healing.
Study Design— Experimental.
Animals— Twelve dogs.
Methods— Paired biopsy samples (ST, HS) were collected from liver, spleen, adrenal gland, pancreas, stomach, jejunum, and bladder using laparoscopic or laparoscopic-assisted methods. Hemorrhage at biopsy sites was assessed (present or absent). Specimens were evaluated for diagnostic quality and histologic changes. Dogs were necropsied at 4 or 14 days postoperatively.
Results— HS incision caused less frequent hemorrhage of all organs except spleen. HS specimens had more coagulation necrosis than ST specimens, but both techniques yielded specimens that were sized similarly and were of diagnostic quality. At necropsy, HS biopsy sites of the stomach, jejunum, bladder, adrenal gland, and pancreas were more hyperemic than ST sites. There were more adhesions at jejunal and pancreatic ST biopsy sites. HS biopsy sites had more coagulation necrosis at days 4 and 14 postoperatively. Differences in fibrin deposition, inflammation, and fibrosis were present at biopsy sites of some organs at days 4 or 14 and in comparisons between days 4 and 14.
Conclusions— Diagnostic quality biopsy specimens were obtained with HS and ST. Although HS-induced gross and histologic changes during the first 2 postoperative weeks, no clinical complications were observed.
Clinical Relevance— Both HS and ST can yield specimens with minimal hemorrhage and HS resulted in no apparent postoperative problems in normal dogs. Although HS caused more inflammation and adhesions at biopsy sites of the pancreas, adrenal gland, and jejunum, no clinical complications occurred.