Dissipation of cyanogenic glucosides and cyanide in soil amended with white clover (Trifolium repens L.)

The use of white clover (Trifolium repens L.) as green manure is common practice due to its high nitrogen content. White clover produces the two cyanogenic glucosides linamarin and lotaustralin, which release toxic hydrogen cyanide upon hydrolysis. The hydrolysis of cyanogenic glucosides and release of cyanide were studied in batch experiments with ground white clover added to soil at loadings of 75 g leaves per kg soil. Linamarin and lotaustralin were quickly hydrolysed with first-order rate constants of 0.026-0.0062 h⁻¹ (corresponding to half lives of 11-27 h) in sandy and loamy soils at natural moisture contents and 11 °C. Experiments with addition of pure cyanogenic glucosides and with sterilized soil material as well as addition of white clover to inert quartz showed that hydrolysis by plant glucosidases is partly inhibited in the soil matrix, but also that soil glucosidases present in soil contribute to degradation of the cyanogenic glucosides, and more so for linamarin than for lotaustralin. Cyanide was produced during the hydrolysis of the cyanogenic glucosides as seen by formation of HCN(g) and WAD-CN (Weak Acid Dissociable Cyanide) amounting to max. 5 and 50% of total-CN in the systems with white clover added to natural soils. However, the increase in WAD-CN was transient, due to subsequent dissipation of the compound caused by abiotic and microbial CN degradation. Due to WAD-CN dissipation in the top soils studied, long term effects of cyanide on sensitive microorganisms and plants are not expected. However, knowledge on the stability of WAD-CN in subsoil is lacking and warrants further investigations.     

Optimal combinations of acute phase proteins for detecting infectious disease in pigs

ABSTRACT: The acute phase protein (APP) response is an early systemic sign of disease, detected as substantial changes in APP serum concentrations and most disease states involving inflammatory reactions give rise to APP responses. To obtain a detailed picture of the general utility of porcine APPs to detect any disease with an inflammatory component seven porcine APPs were analysed in serum sampled at regular intervals in six different experimental challenge groups of pigs, including three bacterial (Actinobacillus pleuropneumoniae, Streptococcus suis, Mycoplasma hyosynoviae), one parasitic (Toxoplasma gondii) and one viral (porcine respiratory and reproductive syndrome virus) infection and one aseptic inflammation. Immunochemical analyses of seven APPs, four positive (C-reactive protein (CRP), haptoglobin (Hp), pig major acute phase protein (pigMAP) and serum amyloid A (SAA)) and three negative (albumin, transthyretin, and apolipoprotein A1 (apoA1)) were performed in the more than 400 serum samples constituting the serum panel. This was followed by advanced statistical treatment of the data using a multi-step procedure which included defining cut-off values and calculating detection probabilities for single APPs and for APP combinations. Combinations of APPs allowed the detection of disease more sensitively than any individual APP and the best three-protein combinations were CRP, apoA1, pigMAP and CRP, apoA1, Hp, respectively, closely followed by the two-protein combinations CRP, pigMAP and apoA1, pigMAP, respectively. For the practical use of such combinations, methodology is described for establishing individual APP threshold values, above which, for any APP in the combination, ongoing infection/inflammation is indicated.     

Embryo culture is an efficient way to conserve a medicinally important endangered forest tree species Strychnos potatorum

The present study reports a protocol for germination of Strychnos potatorum (ver. Tel. Chilla) using zygotic embryo culture as an embryo rescue method. A 100% germination rate was obtained by culturing the embryos on full-strength Murashige and Skoog’s medium (MS) containing 20 g/L sucrose in comparison to McCown and Lloyd’s Woody Plant Medium (WPM). Germination rates decreased when the sucrose concentration was lower or higher than 20 g·L-1 . WPM/MS medium containing glucose at levels 30, 20, 15 g·L-1 showed a smaller percentage of germination and at quarter strength, WPM/MS medium with glucose did not respond. Multiple shoot formation was found at 1.0 2.0 mg/L BAP; 3.0 mg/L Kn; 2.0 mg/L TDZ on MS medium with 20 g·L-1 sucrose. Germination rates improved when the embryos were placed upright (vertically) in the medium. The in vitro germinated seedlings were acclimatized in a walk-in-chamber and maintained in the green house with the survival rate of 65% 75%. These plants were transferred to the field and were found to be phenotypically normal, healthy and similar to donor plants. This protocol will be useful to overcome seed dormancy and for rapid multiplication and conservation of S. potatorum using zygotic embryo culture.     

The negative acute phase response of serum transthyretin following Streptococcus suis infection in the pig

Transthyretin (TTR) is a serum protein which is a negative acute phase reactant in humans and levels of TTR are routinely measured as an indicator of health status. Such tests have yet to be established for the pig. In order to measure serum TTR in the pig during an acute phase response an assay was developed using anti-human TTR antibodies which cross reacted with porcine TTR. The assay had a detection limit of 32 microg/mL while the mean concentration of transthyretin measured in healthy pig serum was 302 +/- 8 microg/mL (n = 63). There was no significant difference in the serum concentration of TTR in three different age groups from 10 to 25 weeks. Following Streptococcus suis type 2 infection transthyretin showed a negative acute phase response with serum concentrations reaching a significantly lower level at two days following infection.     

Mineralization of benzyl glucosinolate and its hydrolysis product the biofumigant benzyl isothiocyanate in soil

Plants produce many secondary metabolites that are used as a defense against herbivores, pests and pathogens. Among them are the glucosinolates (GSLs) that are produced by plants of the Brassicales order. Upon enzymatic hydrolysis, GSLs are transformed to primarily isothiocyanates (ITCs) and nitriles. ITCs are toxic compounds with the potential to be used as biofumigants for the control of soil-borne pests. In this study, mineralization kinetics of benzyl GSL and benzyl ITC in a sandy and a clayey soil at 8–9 °C were investigated. Mineralization of ¹⁴C-benzyl GSL, which was de novo synthesized and purified from transgenic Arabidopsis thaliana after administering l-[U-¹⁴C]phenylalanine, showed that 40–50% was mineralized after 60 days. Mineralization of ¹⁴C-benzyl ITC produced by enzymatic hydrolysis of the ¹⁴C-benzyl GSL resulted in mineralization of 35–50% after 45 days. The ITC or a metabolite of ITC was rate limiting for mineralization of ITC during the first 10–15 days in incubations with subsoils. This effect may be attributed to toxicity of the ITC or an ITC metabolite, or that a lag phase was required for the relevant degraders to grow. Although the compounds are of natural origin and thus “known” to microbial degraders the extent of mineralization was not higher than for synthetic compounds like pesticides. After termination of the mineralization experiments, the soils were extracted with water, ethyl acetate and 4 M NaOH, respectively. These extractions indicated that a large amount of the applied ¹⁴C-labeled benzyl GSL and benzyl ITC was incorporated in organic fractions with low bioavailability.     

Bulk Carbohydrate Grain Filling of Barley β‐Glucan Mutants Studied by 1H HR MAS NMR

Temporal and genotypic differences in bulk carbohydrate accumulation in three barley genotypes differing in the content of mixed linkage β‐(1→3),(1→4)‐D‐glucan (β‐glucan) and starch were investigated using proton high‐resolution, magic angle spinning, nuclear magnetic resonance (¹H HR MAS NMR) during grain filling. For the first time, ¹H HR MAS NMR spectra of flour from immature barley seeds are analyzed. Spectral assignments are made using two‐dimensional (2D) NMR methods. Both α‐ and β‐glucan biosynthesis were characterized by inspection of the spectra as well as by calibration to the reference methods for starch and β‐glucan content. Starch was quantified with very good calibrations to the α‐(1→4) peak (5.29–5.40 ppm) and the region 3.67–3.83 ppm covering starch glycopyranosidic protons from H5 and H6. In contrast, the spectral inspection of the β‐anomeric region 4.45–4.85 ppm showed unexpected lack of intensity in the high β‐glucan mutant lys5f at seed maturity, resulting in poor calibration to reference β‐glucan content. We hypothesize that the lack of β‐glucan signal in lys5f indicates partial immobilization of the β‐glucan that appears to be either genotypic dependent or water/β‐glucan ratio dependent.     

The effects of detomidine, romifidine or acepromazine on echocardiographic measurements and cardiac function in normal horses

OBJECTIVE: To evaluate by echo- and electrocardiography the cardiac effects of sedation with detomidine hydrochloride, romifidine hydrochloride or acepromazine maleate in horses. STUDY DESIGN: An experimental study using a cross-over design without randomization. ANIMALS: Eight clinically normal Standardbred trotters. MATERIALS AND METHODS: Echocardiographic examinations (two-dimensional, guided M-mode and colour Doppler) were recorded on five different days. Heart rate (HR) and standard limb lead electrocardiograms were also obtained. Subsequently, horses were sedated with detomidine (0.01 mg kg(-1)), romifidine (0.04 mg kg(-1)) or acepromazine (0.1 mg kg(-1)) administered intravenously and all examinations repeated. RESULTS: Heart rate before treatment with the three drugs did not differ significantly (p = 0.98). Both detomidine and romifidine induced a significant decrease (p < 0.001) in HR during the first 25 minutes after sedation; while acepromazine had a varying effect on HR. For detomidine, there was a significant increase in LVIDd (left ventricular internal diameter in diastole; p = 0.034) and LVIDs (left ventricular internal diameter in systole; p < 0.001). In addition, a significant decrease was found in IVSs (the interventricular septum in systole; p < 0.001), LVFWs (the left ventricular free wall in systole; p = 0.002) and FS% (fractional shortening; p < 0.001). The frequency of pulmonary regurgitation was increased significantly (p < 0.001). Romifidine induced a significant increase in LVIDs (p < 0.001) and a significant decrease in IVSs (p < 0.001) and FS% (p = 0.002). Acepromazine had no significant effect upon any of the measured values. CONCLUSIONS: and clinical relevance The results indicate that sedation of horses with detomidine and to a lesser extent romifidine at the doses given in this study has a significant effect on heart function, echocardiographic measurements of heart dimensions and the occurrence of valvular regurgitation. Although the clinical significance of these results may be minimal, the potential effects of sedative drugs should be taken into account when echocardiographic variables are interpreted in clinical cases.     

Potential of mixed culture of freshwater prawn (Macrobrachium rosenbergii) and self-recruiting small species mola (Amblypharyngodon mola) in rotational rice–fish/prawn culture systems in Bangladesh

The effects of different stocking densities of freshwater prawn (Macrobrachium rosenbergii) on its growth and production in relation to the presence of small self‐recruiting species mola (Amblypharyngodon mola) were investigated in modified rice fields after rice harvest at Mymensingh, Bangladesh. Keeping the stocking density of mola fixed at 20 000 ha^?1 in each treatment, four densities of freshwater prawn (treatments) were maintained: 10 000, 15 000, 20 000 and 25 000 ha^?1 respectively. The rice plots were limed (CaCO₃) and fertilized with urea, triple super phosphate and cowdung regularly. The prawns were fed daily with commercial pellets. Water quality parameters such as temperature, dissolved oxygen, transparency, pH, total alkalinity, inorganic nitrogen (nitrate, nitrite and total ammonia), chlorophyll a and orthophosphate were determined fortnightly. Numerical analysis of plankton communities was performed monthly. All water quality parameters were found to be within the suitable range for freshwater prawn culture, except high temperature, in the peak summer months. There was a homogenous abundance of plankton communities in all treatments throughout the experimental period. Mola started to breed in the second month of the original stocking with partial harvesting after the second month and continued until the final harvest. The total production of mola ranged between 124 and 152 kg ha^?1 during the 4‐month culture period. The average prawn survival ranged from 49% to 57% without any significant difference among treatments. Freshwater prawn production ranged from 294 to 596 kg ha^?1 with significantly higher production in the treatment where 20 000 ha^?1 freshwater prawn were stocked. This treatment also resulted in a higher net profit margin (74%), indicating that stocking at a combination of 20 000 ha^?1 freshwater prawn and 20 000 ha^?1 mola could be the optimum proposition for prawn–mola culture in modified rain‐fed rice fields after rice cultivation.     

Using tube rhizotrons to measure variation in depth penetration rate among modern North-European winter wheat (Triticum aestivum L.) cultivars

Deeper plant root systems are desired for improved water and nitrogen uptake in leaching environments. However, phenotyping for deep roots requires methods that enable plants to develop deep roots under realistic conditions. Winter cereals raise further complications as early growth occurs under low light and temperature during autumn and winter—conditions not met in standard glasshouse facilities. This study used tube rhizotrons of 2 m length, positioned outdoor under a rainout shelter to screen for depth penetration rates (DPR) of roots. Rooting depths of 1 to 1.5 m were achieved with 23 widely grown North European winter wheat cultivars in two autumn/winter and two summer experiments and nine of the cultivars were represented in two or more experiments. Heritability of DPR of roots was only consistent in autumn/winter experiments (27 %) signifying the importance of phenotyping in relevant seasons and environments. Depth penetration rate of roots varied significantly within the tested cultivars, from 1.39 (±0.35) mm °C^?1 day^?1 for cv. Tuareg to 2.07 (±0.34) mm °C^?1 day^?1, for cv. Mercedes. This study documented consistent differences of DPR among North-European winter wheat cultivars in long tube rhizotrons under semi-natural conditions, which may form part of future phenotyping facilities for deep rooting traits.