A Method for Determining Phosphate Diffusion Coefficients by Bulk Diffusion in Soil

A method is described for the determination of phosphate diffusion coefficients by bulk diffusion in soil using the concentration distance method. Two soil blocks only differing in phosphate concentration are brought into contact. After a diffusion period of two weeks the soil blocks are separated, frozen in liquid nitrogen and sliced into layers about 0.02 cm thick by means of a refrigerated microtome. The soil samples are extracted with 4 N HCl, a procedure which fully recovers the added amount of fertilizer P and thus includes the total amount of P that diffuses from one soil block to the other. A concentration distance profile for P and a calculation of the P diffusion coefficient is presented.     

A method for determining Michaelis-Menten kinetic parameters of nutrient uptake for plants growing in soil

The relationship between nutrient influx (I_n) and solution concentration at the root surface (C_lo) has not yet been determined for roots growing in soil because of difficulties in measuring C_lo. Corn was grown on two soils with 12 and 21% clay. Each soil had five K levels ranging from low to very high. Potassium influx (I_n) was determined from K uptake between two harvests and root length. C_lo was then calculated from the average soil solution concentration and I_n by assuming that diffusion is the main transport mechanism for K to the root. Potassium influx plotted against C_lo showed a saturation curve, typical of a Michaelis-Menten kinetic relationship. The Michaelis-Menten uptake parameters, maximum influx (I_max) and Michaelis constant (K_m), were obtained by the “Hanes” plot. There was close agreement, without lack of fit, between calculated and observed data. The proposed procedure therefore appears to be suitable for estimating the uptake kinetics of roots growing in soil. Requirements for applications of the method are discussed.     

Availability of phosphate and potassium as the result of interactions between root and soil in the rhizosphere

A number of findings are summarized in order to show the significance of individual plant properties and soil factors on the availability of phosphate and potassium to plants growing in soil. The flux of a nutrient into a given plant root depends directly on the concentration of the nutrient in the adjacent solution. In nutrient solution, P and K influx follows Michaelis-Menten kinetics. Almost maximum rates of influx have been observed in the range of soil solution concentrations usually found in German arable soils. Roots exhaust P and K from solutions to about 0.2 μmol P and 1 μmol K 1^?1 if not replenished. At the root surface P and K concentrations in soil decrease rapidly within one day; small changes occur after this period. Initially, the extent of the depletion zone is very small but it extends radially with time. After the initial phase therefore, P and K supply to the plant depends on transport from more remote parts of the soil and also on release from undissolved sources. The degree of depletion and the extent of the depletion zone are related to the diffusion coefficient; they decrease with increasing clay content of soil. Root hairs penetrate the soil and extend the volume of soil supplying nutrients to a unit of root. P and K influx therefore increase with the length of root hairs. Proton release of roots mobilize P and K in soil. This is clearly detected by the HCl-soluble P and K fractions within 2 mm of the root surface. The activity of acid and alkaline phosphatases strongly increase in the soil in the vicinity of the root surface of several plant species. It is supposed that organic P compounds can therefore be utilized by plants. P and K influx per unit of root length and root length per unit of shoot weight differed widely between species. The product of these two parameters however was closely related to the P and K concentration of the shoots. Calculations from a mathematical model were in good agreement with measured K depletion profiles and K uptake by plants. It is therefore concluded that the main factors influencing the P and K availability of plants growing in soil have been accounted for in the mathematical model and that the parameters have been accurately measured.     

Differences in manganese efficiency of wheat (Triticum aestivum L.) and raya (Brassica juncea L.) as related to root‐shoot relations and manganese influx

Manganese efficiency is a term used to describe the ability of plants to obtain higher relative yields at low Mn supply compared to other species. To evaluate Mn efficiency of wheat (Triticum aestivum L.) and raya (Brassica juncea L.), a greenhouse pot experiment was conducted using Mn deficient Typic Ustochrept loamy sand soil, treated with 0, 50, and 100 mg Mn (kg soil)^–1. In the no‐Mn treatment, wheat had produced only 30 % of its maximum dry matter yield (DMY) with a shoot concentration of 10.8 mg Mn (kg DM)^–1 after 51 days of growth, while raya had produced 65 % of its maximum DMY with 13.0 mg Mn (kg DM)^–1. Taking relative shoot yield as a measure of Mn efficiency, raya was more efficient than wheat. Both crops produced the maximum DMY with 50 mg Mn (kg soil)^–1. Even though raya had a lower root length : DMY ratio and a higher shoot growth rate, it acquired higher Mn concentrations in the shoot than wheat under similar soil conditions, because of a 2.5 times higher Mn influx. Model calculations were used to calculate the difference of Mn solution concentration (ΔC_L) between the bulk soil (C_Li) and the root surface (C_L0) that is needed to drive the flux by diffusion equal to the measured influx. The results showed that ΔC_L was smaller than C_Li, which indicates that chemical mobilization of Mn was not needed to explain the observed Mn uptake even for raya. According to these calculations, the higher Mn influx of raya was caused by more efficient uptake kinetics, allowing for a 4.5 times higher Mn influx at the same Mn concentration at the root surface.     

Variants of the recently discovered avian gyrovirus 2 are detected in Southern Brazil and The Netherlands

A genome of a virus preliminarily named avian gyrovirus 2 (AGV2), a close relative to chicken anemia virus, was recently discovered in a chicken in the state of Rio Grande do Sul, Southern Brazil. To study the occurrence of AGV2 in Rio Grande do Sul and the neighboring state Santa Catarina, a number of adult chickens (n=108 and n=48, respectively) were tested for the presence of AGV2 DNA. An AGV2-specific PCR was developed, optimized and used to analyze DNA extracted from clinical samples. AGV2 DNA was detected in 98/108 (90.7%) of samples collected in the state of Rio Grande do Sul and 29/48 (60.4%) of the samples collected in the state of Santa Catarina. In order to check whether AGV2 DNA would be detected in samples from a geographically distant region, DNA from brain samples of 21 diseased chickens from the Netherlands were tested independently, by the same method. In such specimens, 9/21 (42.9%) brain tissue samples were found to contain AVG2 DNA. Sequence analysis of some of the PCR products demonstrated that the amplified AGV2 sequences could vary up to 15.8% and could preliminarily be divided in three groups. This indicated the occurrence of variants of AGV2, which may reflect differences in geographical origin and/or in biological properties. The data presented here provides evidence that AGV2 seems fairly distributed in chickens in Southern Brazil and that AGV2 also circulates in the Netherlands. Besides, circulating viruses display genetic variants whose significance should be further examined, particularly to determine whether AGV2 would play any role in chicken diseases.     

Antigen quantification as in vitro alternative for potency testing of inactivated viral poultry vaccines

Abstract

Routine batch control of licensed inactivated viral vaccines for poultry usually includes a potency assay as a measure of vaccine efficacy. Potency assays often consist of vaccination‐challenge experiments in the target species or in laboratory animals. Instead of measuring the protection of vaccinated animals against virulent pathogens, the serological response after vaccination can be quantified for some vaccines. In vitro antigen quantification assays would be attractive alternatives for the current potency assays because the time and costs involved could be greatly reduced and animal use could be avoided. Such in vitro assays will only be acceptable when the correlation between results and efficacy or potency has been demonstrated convincingly.

The results of our studies on antigen quantification assays indicate that, in principle, quantification of viral antigens from inactivated oil‐adjuvanted vaccines is feasible and reproducible using specially developed antigen capture ELISAs in combination with specific software for statistical analysis of the ELISA data. We have developed methods to quantify the haemagglutination‐neuraminidase (HN) and fusion (F) proteins of Newcastle disease virus (NDV), the viral protein 3 (VP3) of the infectious bursal disease virus (IBDV), and the spike‐1 (S1) protein of the infectious bronchitis virus (IBV). Vaccination experiments with inactivated ND vaccines indicate that the in vitro quantified HN‐ or F‐proteins of NDV are reliable indicators of the serological response after vaccination.     

Longevity of Mycobacterium bovis in brushtail possum (Trichosurus vulpecula) carcasses,and contact rates between possums and carcasses

Abstract

AIM: To determine, for a variety of environmental conditions, how long Mycobacterium bovis might remain viable inside the carcass of a brushtail possum (Trichosurus vulpecula) that died of bovine tuberculosis (Tb), and to measure the rate of contact between free-ranging possums and possum carcasses.

METHODS: Lesions of M. bovis were simulated by inoculating excised spleens weighing 0.5–1 g with 0.2 mL liquid culture containing approximately 5 x 10⁷ cfu M. bovis/mL. Simulated lesions were inserted into possum carcasses (n=48) at the peripheral lymph nodes. Carcasses were placed in the field at two sites (a tussock grassland and a podocarp-broadleaved forest site) and in two seasons (summer and winter) for up to 62 days. Survival rates of M. bovis were estimated by sampling the simulated lesions over time, and culturing the recovered lesion to determine if any viable M. bovis bacteria were present.

The time taken for a free-ranging possum to first encounter a dead possum in its home range was estimated by live-trapping possums and fitting them with proximity loggers (n=13). A ‘contact’ was recorded if these possums came within 40–50 cm of proximity loggers fitted to possum carcasses.

RESULTS: There were strong seasonal and site effects in the survival rate of M. bovis in possum carcasses. In the grassland habitat, no viable bacilli were cultured from any carcass after 3 days in summer, whereas in winter all samples were culture-positive for the first 20 days, and some were still positive after 27 days. The survival rates for forest habitat were intermediate between the results for grassland, and there were no culture-positive carcasses after 9 days in summer or 27 days in winter.

In summer, infected carcasses (n=6) were first encountered by possums a mean 1.9 (range 0.4–6.7) days after placement.

CONCLUSIONS: Possum carcasses were contacted by free-ranging possums within the period that viable M. bovis were shown to survive in a carcass. The risk of such infection is likely to be most significant in winter or in areas with microhabitats where the survival of M. bovis is high. However, the generally low survival rate of M. bovis in possum carcasses and the low frequency of possum-to-carcass contacts indicate this route of transmission alone could not maintain Tb in a possum population.     

A Bayesian meta-analysis of the effects of administering an intra-vaginal (CIDR) device in combination with other hormones on the reproductive performance of cycling,anoestrous and inseminated cows

AIMS: To evaluate the effectiveness of treatment programmes that included controlled internal drug-releasing (CIDR) devices containing progesterone (P4) in improving synchrony of oe- strus, and conception and pregnancy rates in cycling, anoestrous and inseminated dairy cows, using meta-analysis. To describe the difference in response between cycling and anoestrous cows to CIDR-based synchrony programmes.

METHODS: Scientific papers written in the English language between 1989 and 2002 that investigated the effects of treat- ment programmes including CIDR devices on reproductive per- formance in dairy heifers or lactating dairy cows were identified using a computerised literature search. The criteria for inclusion incorporated evidence that treatment allocation was completely randomised; the population studied was lactating dairy cows; and that data were available on submission, conception and pregnancy rates and their associated measures of variability. Re- productive outcomes from 25 synchrony trials (total n= 11,058 cows) were analysed. Summary measures of the effect of treat- ment on reproductive outcome were assessed using fixed- and random-effects Bayesian meta-analysis models.

RESULTS: Treatment programmes including a CIDR device increased the risk of submission in cycling cows (predicted Bayesian RR=2.86, 95% credible interval 1.46–5.67). Compared with controls, synchrony programmes including CIDR devices in cycling dairy cows had no effect on the risk of conception to first service post-treatment (predicted Bayesian RR=1.00, 95% credible interval=0.80-1.24). Compared with controls, synchrony programmes including CIDR devices had no effect on the risk of pregnancy throughout the mating period (pre- dicted Bayesian RR=1.02, 95% credible interval=0.89-1.17). In anoestrous cows, CIDR treatment had no effect on the risk of conception to first service post-treatment and no effect on the risk of pregnancy throughout the mating period, compared with anoestrous, untreated controls (predicted Bayesian RR=0.91 and 0.97, respectively; 95% credible interval=0.68–1.26 and 0.59-1.60, respectively).

CONCLUSION: The results of this meta-analysis showed that synchrony programmes using CIDR devices combined with other hormones reliably enhanced submission rates in lactat- ing dairy cows. The relatively small number of trials with data suitable for analysis and the heterogeneity of results at the indi- vidual trial level limited our ability to confirm either a beneficial or deleterious effect of treatment on conception or pregnancy rates. Further randomised, controlled trials to evaluate the ef- fectiveness of this form of reproductive therapy in commercial dairy farms are needed.