Indigofera spicata (creeping indigo) poisoning of three ponies

Three ponies continuously grazed a pasture containing an estimated 24% Indigofera spicata (wet weight basis) for 4–6 weeks in April and May 2004. They developed ataxia, paresis, depression, muscle fasciculations, dysphagia, ptyalism and halitosis. Two also developed corneal opacity. One pony recovered with supportive treatment, but the other two were euthanased and necropsied. Neuropathology was not present in either case, but both livers had periacinar and periportal lymphocytic infiltrations and hydropic degeneration of mid‐zonal hepatocytes, with mild to moderate periacinar necrosis also evident in one. The I. spicata contained 2.66 mg 3‐nitropropionic acid (3‐NPA)/g dry matter and 1.5 mg indospicine/g dry matter. Indospicine, but not 3‐NPA, was detected in serum from both of the euthanased ponies and indospicine was detected in heart, liver and muscle from the one pony in which this assay was performed. The clinical syndrome closely resembled ‘Birdsville horse disease’ caused by I. linnaei and was similar to that reported in horses poisoned by the closely related species I. hendecaphylla and to 3‐NPA poisoning of other animals, including humans. 3‐NPA is thought to cause this neurological syndrome. To our knowledge, this is the first authenticated report of I. spicata poisoning in grazing animals. We also report here the first published evidence that 3‐NPA and indospicine exist in naturalised I. spicata in Australia and of the formation of indospicine residues in tissues of animals grazing paddocks infested with I. spicata.     

Conjunctival vaccination of the brushtail possum (Trichosurus vulpecula) with bacille Calmette-Guérin

AIM: To determine the efficacy of conjunctival vaccination of captive brushtail possums (Trichosurus vulpecula) with bacille Calmette-Guérin (BCG), as measured by immunological responses to vaccination and response to intratracheal challenge with Mycobacterium bovis.

METHODS: Nine adult male brushtail possums were vaccinated by the instillation of a suspension of BCG strain Pasteur 1173P2 into the conjunctival sac of each eye. Each drop contained approximately 2.5 × 10⁵ colony forming units (cfu). At 8 weeks post-vaccination (pv) the vaccinated possums and 10 unvaccinated possums were challenged by intratracheal instillation of approximately 100 cfu of M. bovis. Cellular immune responses to bovine purified protein derivative (PPD) antigen were measured using the lymphocyte proliferation assay (LPA). Possums surviving to 50–51 days after challenge were euth anised and subjected to detailed post-mortem examination, including histopathology, to assess protection against tuberculosis. Sections of lung and spleen were cultured for M. bovis.

RESULTS: No conjunctival inflammation or other adverse reactions to the administration of the vaccine were evident macroscopically. The vaccinated group showed a systemic cellular immune response to bovine PPD antigen at 4 and 8 weeks pv, and the response at 8 weeks was significantly greater than at 4 weeks (p<0.05). Conjunctival vaccination induced significant levels of protective immunity, measured as less mass of tuberculous lesions in lung (p<0.05) and less dissemination of disease in vaccinated compared with unvaccinated possums (p<0.05).

CONCLUSIONS: Conjunctival vaccination with BCG induced a significant level of protective immunity against M. bovis infection in possums. This route of vaccination, together with intranasal aerosol vaccination, could be utilised in the delivery of an aerosolised vaccine using a device that sprays the vaccine suspension into the eyes and nose of possums.     

Assessment of leaf cover and crop soil cover in weed harrowing research using digital images

Objective assessment of crop soil cover, defined as the percentage of leaf cover that has been buried in soil because of weed harrowing, is crucial to further progress in post‐emergence weed harrowing research. Up to now, crop soil cover has been assessed by visual scores, which are biased and context‐dependent. The aim of this study was to investigate whether digital image analysis is a feasible method to estimate crop soil cover in the early growth stages of cereals. Two main questions were examined: (i) how to capture suitable digital images under field conditions with a standard high‐resolution digital camera and (ii) how to analyse the images with an automated digital image analysis procedure. The importance of light conditions, camera angle, size of recorded area, growth stage and direction of harrowing were investigated, in order to establish a standard for image capture and an automated image analysis procedure based on the excess green colour index was developed. The study shows that the automated digital image analysis procedure provided reliable estimations of leaf cover, defined as the proportion of pixels in digital images determined to be green, which were used to estimate crop soil cover. A standard for image capture is suggested and it is recommended that digital image analysis be used to estimate crop soil cover in future research. The prospects of using digital image analysis in future weed harrowing research are discussed.     

Expert opinion as 'validation' of risk assessment applied to calf welfare

Background

Recently, a Risk Assessment methodology was applied to animal welfare issues in a report of the European Food Safety Authority (EFSA) on intensively housed calves.

Methods

Because this is a new and potentially influential approach to derive conclusions on animal welfare issues, a so-called semantic-modelling type ''validation'' study was conducted by asking expert scientists, who had been involved or quoted in the report, to give welfare scores for housing systems and for welfare hazards.

Results

Kendall''s coefficient of concordance among experts (n = 24) was highly significant (P < 0.001), but low (0.29 and 0.18 for housing systems and hazards respectively). Overall correlations with EFSA scores were significant only for experts with a veterinary or mixed (veterinary and applied ethological) background. Significant differences in welfare scores were found between housing systems, between hazards, and between experts with different backgrounds. For example, veterinarians gave higher overall welfare scores for housing systems than ethologists did, probably reflecting a difference in their perception of animal welfare.Systems with the lowest scores were veal calves kept individually in so-called "baby boxes" (veal crates) or in small groups, and feedlots. A suckler herd on pasture was rated as the best for calf welfare. The main hazards were related to underfeeding, inadequate colostrum intake, poor stockperson education, insufficient space, inadequate roughage, iron deficiency, inadequate ventilation, poor floor conditions and no bedding. Points for improvement of the Risk Assessment applied to animal welfare include linking information, reporting uncertainty and transparency about underlying values.

Conclusion

The study provides novel information on expert opinion in relation to calf welfare and shows that Risk Assessment applied to animal welfare can benefit from a semantic modelling approach.     

Co‐culture of Buffalo Preantral Follicles with Different Somatic Cells

The effect of co‐culture of buffalo preantral follicles (PFs) with different somatic cells, i.e, cumulus, granulosa, ovarian mesenchymal and oviductal epithelial cells was studied. Large PFs (250–450 μm) were isolated by microdissecting the trypsin (1%) digested ovarian cortical slices. Cumulus cells were isolated by repeated pipetting of oocytes, granulosa cells were isolated by aspirating from punctured PFs and ovarian mesenchymal cells were isolated from ovarian cortex by scraping the cortical slices and passing through 20 μm filter. Preantral follicles were cultured in standard culture medium without somatic cells or co‐cultured with cumulus cells, granulosa cells, ovarian mesenchymal cells and oviductal epithelial cells for 80 days. The growth rate (μm/day) of the PFs was monitored by measuring follicular diameter on day 0, 30, 60 and 80 days of culture. The viability of PFs was evaluated by trypan blue staining. The results indicated that PFs co‐cultured with cumulus, granulosa and ovarian mesenchymal cells had a better development and survivality compared with control and those co‐culture with oviductal epithelial cells. Maximum growth and survivality of PFs were achieved when cultured with cumulus cells. It is concluded that inclusion of somatic cells in PF culture media had beneficial effect on the growth of PFs and cumulus cells supported maximum growth and survivality of PFs in vitro of all somatic cells tested.     

土壤微量节肢动物组织中镉测定方法的改进

Highly precise and reliable determination of heavy metals in soil micro-arthropod tissues remains a challenge because of the small size of the animals and their typical low abundance in metal-contaminated agricultural soils. The present study sought to develop a method for cadmium(Cd) determination in soil micro-arthropods by optimizing the sample digestion procedure, reducing sample weight, modifying sample pre-treatment and validating the methodology with field samples. The optimized digestion conditions comprised a sample mass of 50–150 μg, digestion reagent of nitric acid:hydrogen peroxide(3:1), digestion temperature of 105℃,digestion period of 3 h and digestion volume of 30 μL. Defecation of the standard Collembola Folsomia candida(92 h) and the indigenous Collembola Onychiurus yodai(42 h) and ultrasonic cleaning of F. candida increased the accuracy of Cd determination.The recovery of Cd using the refined procedure was 98.9% and the limits of detection and quantification were 0.002 and 0.008 μg L~(-1), respectively. The within-batch precision values were 3%. The Cd concentrations in the tissues of the Collembola Isotoma sp.collected from a range of metal-contaminated fields determined by the improved method were consistent with the Cd concentrations in the field soils. The results indicate that the optimized method can be used for more accurate or reliable determination of Cd concentrations in soil micro-arthropod tissues.     

Evidence for niche differentiation of nitrifying communities in grassland soils after 44 years of different field fertilization scenarios

Long-term nitrogen(N)fertilization imposes strong selection on nitrifying communities in agricultural soil,but how a progressively changing niche affects potentially active nitrifiers in the field remains poorly understood.Using a 44-year grassland fertilization experiment,we investigated community shifts of active nitrifiers by DNA-based stable isotope probing(SIP)of field soils that received no fertilization(CK),high levels of organic cattle manure(HC),and chemical N fertilization(CF).Incubation of DNA-SIP microcosms showed significant nitrification activities in CF and HC soils,whereas no activity occurred in CK soils.The 44 years of inorganic N fertilization selected only 13C-ammonia-oxidizing bacteria(AOB),whereas cattle slurry applications created a niche in which both ammonia-oxidizing archaea(AOA)and AOB could be actively13C-labeled.Phylogenetic analysis indicated that Nitrosospira sp.62-like AOB dominated inorganically fertilized CF soils,while Nitrosospira sp.41-like AOB were abundant in organically fertilized HC soils.The 13C-AOA in HC soils were affiliated with the 29i4 lineage.The 13C-nitrite-oxidizing bacteria(NOB)were dominated by both Nitrospira-and Nitrobacter-like communities in CF soils,and the latter was overwhelmingly abundant in HC soils.The 13C-labeled nitrifying communities in SIP microcosms of CF and HC soils were largely similar to those predominant under field conditions.These results provide direct evidence for a strong selection of distinctly active nitrifiers after 44 years of different fertilization regimes in the field.Our findings imply that niche differentiation of nitrifying communities could be assessed as a net result of microbial adaption over 44 years to inorganic and organic N fertilization in the field,where distinct nitrifiers have been shaped by intensified anthropogenic N input.     

接种丛枝菌根真菌和巨大芽孢杆菌的红三叶草对难溶磷的利用

A pot experiment was conducted to investigate the mobilization of sparingly soluble inorganic andorganic sources of phosphorus (P) by red clover (Trghlium pmtense L.) whose roots were colonized by the arbuscular mycorrhizal (AM) fungus Glomus mosseae and in association with the phosphate-solubilizing (PS) bacterium Bacillus megaterium ACCC10010. Phosphate-solubilizing bacteria and rock phosphate hada synergistic effect on the colonization of plant roots by the AM fungus. There was a positive interaction between the PS bacterium and the AM fungus in mobilization of rock phosphate, leading to improved plant P nutrition. In dual inoculation with the AM fungus and the PS bacterium, the main contribution to plant P nutrition was made by the AM fungus. Application of P to the low P soil increased phosphatase activityin the rhizosphere. Alkaline phosphatase activity was significantly promoted by inoculation with either the PS bacterium or the AM fungus.