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.     

Use of soil moisture data for refined GreenSeeker sensor based nitrogen recommendations in winter wheat (Triticum aestivum L.)

Previous studies have shown the importance of soil moisture (SM) in estimating crop yield potential (YP). The sensor based nitrogen (N) rate calculator (SBNRC) developed by Oklahoma State University utilizes the Normalized Difference Vegetation Index (NDVI) and the in-season estimated yield (INSEY) as the estimate of biomass to assess YP and to generate N recommendations based on estimated crop need. The objective was to investigate whether including the SM parameter into SBNRC could help to increase the accuracy of YP prediction and improve N rate recommendations. Two experimental sites (Lahoma and Perkins) in Oklahoma were established in 2006/07 and 2007/08. Wheat spectral reflectance was measured using a GreenSeeker? 505 hand-held optical sensor (N-Tech Industries, Ukiah, CA). Soil–water content measured with matric potential 229-L sensors (Campbell Scientific, Logan, UT) was used to determine volumetric water content and fractional water index. The relationships between NDVI, INSEY and SM indices at planting and sensing at 5, 25, 60 and 75-cm depths versus grain yield (GY) were evaluated. Wheat GY, NDVI at Feekes 5 and soil WC at planting and as sensed at three depths were also analyzed for eight consecutive growing seasons (1999–2006) for Lahoma. Incorporation of SM into NDVI and INSEY calculations resulted in equally good prediction of wheat GY for all site-years. This indicates that NDVI alone was able to account for the lack of SM information and thus lower crop YP. Soil moisture data, especially at the time of sensing at the 5-cm depth could assist in refining winter wheat YP prediction.     

Automatic corn plant location and spacing measurement using laser line-scan technique

Identifying corn plant location and/or spacing is important for predicting yield potential and making decisions for in-season nitrogen application rate. In this study, an automatic corn stalk identification system based on a laser line-scan technique was developed to measure stalk locations during corn mid-growth stages. A laser line-scan technique is advantageous in this application because the line-scan data sets taken from various points of view of a plant stalk results in less interference and higher probability of plant recognition. Data were collected for two 10-meter-long corn rows at the growth stages of V8 and V10 using a mobile test platform in 2011. Each potential stalk cluster was identified in a scan and registered with the same stalks in previous scans. The final location of a stalk was the average of the measured locations in all scans. The current system setup with data processing algorithms achieved 24.0 and 10.0 % of mean total errors in plant counting at the V8 and V10 growth stages, respectively. The root-mean-squared error (RMSE) between system measured plant locations and manually measured ones were 2.3 and 2.6 cm at the V8 and V10 growth stages, respectively. The interplant spacing measured by the developed system had a good correlation with the manual measurement with an R ² of 0.962 and 0.951 for the V8 and V10 growth stages, respectively. This system can be ultimately integrated in a variable-rate-spraying system to improve real-time, high spatial resolution variable-rate nitrogen applications.     

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.     

Metabolism of propionate and 1,2-propanediol absorbed from the washed reticulorumen of lactating cows

To investigate the metabolism of 1,2-propanediol (PPD) in lactating cows independently of normal rumen microbial metabolism, three ruminally cannulated lactating Holstein cows were subjected to three experimental infusion protocols under washed reticulo-ruminal conditions in a Latin square design. Reticulo-ruminal absorption rates were maintained for 420 min by continuous intraruminal infusion of VFA and PPD. With the control treatment, 1,246 +/- 39 mmol/ h of acetate and 213 +/- 5 mmol/h of butyrate were absorbed from the reticulorumen. With the propionate treatment, 1,148 +/- 39 mmo/h of acetate, 730 +/- 23 mmol/h of propionate and 196 +/- 5 mmol/h of butyrate were absorbed from the reticulorumen. With PPD treatment, 1,264 +/- 39 mmol/h of acetate, 220 +/- 5 mmol/h of butyrate and 721 +/- 17 mmol/h of PPD were absorbed from the reticulorumen. Glucose irreversible loss rate (ILR), as well as the relative enrichment of plasma lactate and alanine, were determined by primed continuous infusion of [U-13C]glucose in a jugular vein. Treatments did not affect (P > 0.10) the plasma concentrations of glucose (4.2 +/- 0.1 mmoVL), alanine (0.14 +/- 0.01 mmol/L), or insulin (80 +/- 25 pmol/L). The plasma concentration of lactate was higher (P < 0.05) with both propionate (0.84 +/- 5 mmol/L) and PPD treatment (0.81 +/- 5 mmol/ L) compared with the control treatment (0.29 +/- 0.5 mmol/L). The plasma concentration of pyruvate was higher (P < 0.05) with the propionate treatment (0.09 +/- 0.01 mmol/L) compared with the control treatment (0.03 +/- 0.01 mmol/L). The plasma concentration of 3-hydroxybutyrate was lower (P < 0.05) with the propionate treatment (0.15 +/- 0.03 mmol/L) compared with the control treatment (0.40 +/- 0.03). With the PPD treatment, the plasma concentrations of pyruvate and 3-hydroxybutyrate were in between the other treatments and tended (P < 0.10) to be different from both. The plasma concentration of PPD increased throughout the infusion period with the PPD treatment and reached a concentration of 4.9 +/- 0.6 mmol/L at 420 min. The ILR of glucose was not affected (P > 0.10) by treatments (441 +/- 35 mmol/h). The relative 13C enrichment of plasma lactate compared with that of glucose decreased (P < 0.05) with the PPD treatment compared with the control treatment (44 to 21 +/- 3%). It was concluded that PPD has a low rate of metabolism in cows without a normal functioning rumen, although about 10% of the absorbed PPD was metabolized into lactate.     

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.     

Nitrogen fertilizer recommendations based on plant sensing and Bayesian updating

Methods are available to predict nitrogen needs of winter wheat based on plant sensing, but adoption rates by producers are low. Current algorithms that provide nitrogen recommendations based on plant sensing implicitly assume that parameters are estimated without error. A Bayesian updating method was developed that can incorporate precision plant sensing information and is simple enough that it could be computed on-the-go. The method can consider producers prior information and can account for parameter uncertainty. Bayesian updating gives higher nitrogen recommendations than plant sensing recommendations using a plug-in method. These recommendations increase net returns over the previous recommendations, but not enough to make plant sensing profitable in this scenario.     

Effect of nitrogen fertilizer source on corn (Zea mays L.) optical sensor response index values in a rain-fed environment

The use of optical sensors to detect nitrogen (N) deficiencies and determine in-season fertilizer recommendations has grown. Nitrogen responses are difficult to detect early in the growing season. The objective of this experiment was to determine if different N sources could deliver early season detection of N deficiencies. Four N fertilizer sources were applied at rates of 90 and 180 kg N ha^?1 across three site-years. A Greenseeker and SPAD sensor were used to measure in-season fertilizer response index (RI). When differences in sensor RI values between N rates were present, they did not occur until the V9/10 growth stage. No specific N source provided superior results that led to a reliable, early season detection of N deficiency. Reliable differences in response index values could be detected beyond the V7/V8 growth stages. For earlier detection of N responsiveness, and potential N deficiency, other management strategies should be investigated.