Effects of processing and of storage on the stability of pantothenic acid in sea buckthorn products (Hippophaë rhamnoides L. ssp. rhamnoides) assessed by stable isotope dilution assay

A stable isotope dilution assay for quantification of pantothenic acid in sea buckthorn berries, juice, and concentrate using a four-fold labeled isotopologue of vitamin B5 as the internal standard was adopted using reversed phase liquid chromatography-mass spectrometry with electrospray ionization. Because of a rapid sample clean up procedure without the necessity of external calibration, this methodology permits the accurate analysis of a high number of samples within a short time. Sea buckthorn juice was stored at 25 and 40 degrees C for up to 7 days to determine the effects of storage temperature on the stability of pantothenic acid. Analysis of kinetic data suggested that the degradation follows a first-order model. The results of the experiments showed that storage of sea buckthorn juice for 7 days at ambient temperature (25 degrees C) already resulted in a significant degradation of pantothenic acid of about 18%. The processing effects of juice production and subsequent concentration revealed a decrease of about 6-7% in the juice and of 23% in the juice concentrate.     

Effects of past and current crop management on soil microbial biomass and activity

Because soil biota is influenced by a number of factors, including land use and management techniques, changing management practices could have significant effects on the soil microbial properties and processes. An experiment was conducted to investigate differences in soil microbiological properties caused by long- and short-term management practices. Intact monolith lysimeters (0.2 m² surface area) were taken from two sites of the same soil type that had been under long-term organic or conventional crop management and were then subjected to the same 2.5-year crop rotation [winter barley (Hordeum vulgare L.), maize (Zea mais L.), lupin (Lupinus angustifolius L.), and rape (Brassica napus L. ssp. oleifera)] and two fertilizer regimes (following common organic and conventional practices). Soil samples were taken after crop harvest and analyzed for microbial biomass C and N, microbial activity (fluorescein diacetate hydrolysis, arginine deaminase activity, and dehydrogenase activity), and total C and N. The incorporation of the green manure stimulated growth and activity of the microbial communities in soils of both management histories. Soil microbial properties did not show any differences between organically and conventionally fertilized soils, indicating that crop rotation and plant type had a larger influence on the microbial biomass and enzyme activities than fertilization. Initial differences in microbial biomass declined, while the effects of farm management history were still evident in enzyme activities and total C and N. Links between enzyme activities and microbial biomass C varied depending on treatment, indicating differences in microbial community composition.     

Animal treading stimulates denitrification in soil under pasture

The effects of animal treading on denitrification in a mixed ryegrass-clover pasture were studied. A single treading event of moderate or severe intensity was applied in plots during spring by using dairy cows at varying stocking rates (4.5 cows 100 m⁻² for 1.5 or 2.5 h, respectively). Treading caused a significant short-term 21 days) increase in denitrification. Denitrification rates reached a maximum of 52 g N₂O-N ha⁻¹ day⁻¹ at 8 days after severe treading compared to 2.3 g N₂O-N ha⁻¹ day⁻¹ under nil treading. Thereafter, denitrification rates declined, and were similar to non-trodden control plots after 28 days. Soil aeration, was significantly reduced by treading as expressed by water-filled porosity. In addition, soil NH₄⁺-N and NO₃⁻-N concentrations were also increased by treading. We propose that the underlying processes involved in increasing denitrification under treading were two-fold. Firstly, treading caused a temporary (e.g. 3 days after treading) reduction in soil aeration through soil physical damage, and secondly, reduced soil N utilisation prompted by reduced plant growth led to increased soil NH₄⁺-N and NO₃⁻-N availability. This study shows that treading, without the influence of other grazing animal factors (e.g. excretion), can cause a large short-term stimulation of denitrification in grass-clover pastures.     

Cool Season C3-Grasses (Wheat,Rye, Barley,and Oats) Differ in Shoot: Root Ratio When Applied With Different NPK Sources

Shoot:root (S:R) response in four winter C₃-grasses (cereals) viz. wheat (Triticum aestivum L.), rye (Secale cereale L.), barley (Hordeum vulgare L.) and oats (Avena sativa L.) was investigated under eight different NPK sources (S₁ = 20-20-20, S₂ = 20-27-5, S₃ = 7-22-8, S₄ = 10-10-10-20S, S₅ = 11-15-11, S₆ = 31-11-11, S₇ = 24-8-16, and S₈ = 19-6-12) in pot experiment at Dryland Agriculture Institute, West Texas A&M University, Canyon, Texas, USA during winter 2009–2010. The experiment was performed in completely randomized design (CRD) with three repeats. The objective of this experiment was to investigate whether the S:R of winter cereals differ or not when applied with different NPK sources. Considerable variations in the shoot: root was noticed in the four grasses at different growth stages, i.e., 30, 60 and 90 days after emergence (DAE) when applied with different NPK fertilizers sources. Wheat had the highest S:R of 3.4 at 30 days after emergence (DAE), barley at 60 DAE (6.5), while oats at 90 DAE (3.9). Among the NPK sources, crops had the highest S:R of 3.6, 6.2 and 3.7 when applied with S₃, S₂, and S₆ at 30, 60 and 90 DAE, respectively. Shoot to root ratio showed positive relationship with increase in shoot weight and negative relationship with increase in root weight. The NPK fertilizer S₆ (31: 11: 11), an acid loving fertilizer had negative effects on the shoot and root development of cool season cereals, but among these cereals under study, barley and oats roots were affected more than wheat and rye. It was concluded from this present study, that the four cool season cereals responded differently in terms of shoot: root to different NPK fertilizers at different growth stages.     

A duplex-PCR bioassay to detect a Trichoderma virens biocontrol isolate in non-sterile soil

This paper describes the identification and utilisation of a sequence-characterised amplified region (SCAR) marker specific for the Trichoderma virens biocontrol isolate GV4. The marker was developed from a RAPD-PCR amplification product unique to isolate GV4. When used as a hybridisation probe in Southern blot analysis, it hybridised to the DNA of the species T. virens alone and not to that of other Trichoderma species or closely related genera Gliocladium and Verticillium. The marker also produced a GV4-specific RFLP, distinguishing it from other T. virens isolates when probed to blots with HindII, BamHI or PstI genomic DNA digests. Primers designed from the sequence of the RAPD marker produced a diagnostic amplification product of 346 bp for GV4 alone, distinguishing it from all other test isolates. With the exception of one, test isolates did not produce an amplification product with the SCAR primers. The exception was a single Verticillium psalliotae isolate (ICMP5509) that produced a product of approx. 400 bp that was easily distinguished from the 346 bp product of GV4. The reliability of the SCAR-based diagnostic test was further improved with the introduction of a positive PCR reaction control to each test, achieved by converting the test to a duplex PCR system. Two universal primers flanking the two ITS and the 5.8S region of the ribosomal gene complex were introduced to each reaction to provide a test for PCR reaction inhibitors to eliminate false negatives in the diagnosis. Amplification of this multi-copy genomic region did not reduce diagnostic sensitivity of the single copy SCAR marker. To further increase the sensitivity of detecting GV4 propagules while maintaining a fast sample assessment assay, soil was amended with cornmeal, as a nutrient source, and a mix of antibiotics to favour Trichoderma growth. The soil mix was subsequently incubated for 5 d before total DNA was extracted. Under these conditions, the duplex soil PCR assay detected GV4 down to a concentration of 10 spores g⁻¹ soil in non-sterile agricultural field soil. This study is the first to report the use of a duplex-PCR diagnostic bioassay for a species within the Hypocrea/Trichoderma genus.     

Coccidia of the llama, Lama glama, in Colorado and Wyoming.

Fecal samples were collected from 144 llamas, Lama glama, on four premises in northern Colorado and southern Wyoming. Feces were examined for the presence of coccidial oocysts; Eimeria alpacae was found in 55.6%, Eimeria lamae in 67.3% and Eimeria macusaniensis in 1.4% of the samples. An adult female llama was found to be infected with Eimeria macusaniensis at necropsy. This is the first report of these species of coccidia in the llama.     

Use of computed tomography to evaluate pathologic changes in the lungs of calves with experimentally induced respiratory tract disease

OBJECTIVE: To optimize methods for the use of computed tomography (CT) to assess pathologic changes in the lungs of calves and to determine the effect of treatment on lung consolidation. ANIMALS: 10 male Holstein calves. PROCEDURES: Calves were anesthetized to facilitate CT imaging of the thorax. After initial images were obtained, pneumonia was induced in the calves by inoculation through a bronchoscope. Two calves were used in a preliminary study to refine the inoculation dose and optimize CT images. Four calves were administered florfenicol and 4 calves were untreated control animals. Serial images were obtained 24, 48, and 72 hours after inoculation. After final images were obtained, calves were euthanized, and lung consolidation was estimated by use of lung surface area scoring and water displacement. These estimates were compared with estimated lung consolidation obtained by use of CT. RESULTS: Calves had rapid disease progression. Percentage of lung consolidation was not significantly different between treatment groups for any of the estimation methods. Results of an ANOVA of the 3 assessment methods indicated significant differences among methods. Estimates of the percentage of lung consolidation obtained by use of surface area scoring and CT correlated well, whereas water displacement estimates correlated poorly with other methods of consolidation estimation. CONCLUSIONS AND CLINICAL RELEVANCE: Because of the correlation with other methods for estimation of lung consolidation, CT has the potential to be used to monitor disease progression in calves with experimentally induced respiratory tract disease.