Degradation of ricin in castor seed meal by temperature and chemical treatment

Oil from the seed of the castor plant (Ricinus communis L.) is an important commodity for a number of industries, ranging from pharmaceuticals to renewable energy resources. However, the seed and subsequent seed meal contain ricin (RCA₆₀), a potent cytotoxin, making it an unusable product. In order to investigate the efficiency of previously researched methods of reducing the toxicity of the meal, cold-pressed oil extracted seed meal known to contain ricin was boiled in the presence of 50 mM calcium hydroxide (pH 12.5). However, boiling of this seed meal in the presence of calcium hydroxide produced no significant difference from boiling alone. Therefore, heat and chemical treatments were performed to determine their effects on the denaturation of the ricin within whole seed, milled un-extracted seed, and cold-pressed extracted seed. Boiling and autoclaving showed varying degrees of effectiveness depending on the sample type. Ricin within the cold-pressed extracted meal was rendered unresponsive to antibody probing after 10 min of boiling or autoclaving. In contrast, treatment of cold-pressed extracted meal with 8 M urea and 6 M guanidine–HCl for 60 min produced no observable reduction in the response of the ricin to the antibody. Critically, hot pressing of the castor seed produced meal that exhibited no reactivity with the antibody, indicating that the ricin had been denatured during the oil extraction. By removing the toxic component of the castor meal, this by-product could create a new commodity from the production of castor oil, thereby making castor oil production more profitable.     

Agroforestry: southern pines and subterranean clover cultural treatments

Agroforestry research in central Louisiana, USA, examined the effects of agronomic cultural treatments (disk, chemical, hay, and hay-graze) on subterranean clover (Trifolium subterraneum L.) yields and slash pine (Pinus elliottii Engelm.) and loblolly pine (P. taeda L.) survival and growth. Cultural treatments appear necessary for successful production of subterranean clover in tame pastures, especially for early fall forage growth. Subterranean clover yields in volunteer swards during spring were highest on the disk and chemical treatments. Generally, pine survival and growth during the first 5 years following tree establishment were not affected by cultural treatments.     

Equine herpesvirus 2-associated granulomatous dermatitis in a horse

Granulomatous dermatitis in horses has been linked to many etiologies, including various parasites, fungi, and bacteria. Idiopathic forms of granulomatous inflammation-producing diseases, some of which are localized to the skin, also have been reported in horses. Herein we describe a case of recurrent equine granulomatous skin disease characterized by intranuclear viral inclusions within macrophages and giant cells. The histologic changes were primarily noted in the deep dermis and included multifocal to coalescing areas of necrosis marked by histiocytic cell infiltration and presence of giant cells. Electron microscopic examination revealed intranuclear and intracytoplasmic viral particles consistent with herpesvirus. Sequence results of the polymerase chain reaction product were consistent with equine herpesvirus 2, adding another possible etiology to the list of differentials in cases of equine granulomatous skin disease.     

A demographic survey of unwanted horses in Ireland 2005-2010

Background

The Irish Horse Industry expanded during the Celtic Tiger boom years, then contracted in the current economic recession. High value horses were traditionally controlled through sale at public auction, private sales and sales to dealers; these are now also being reduced by decreases in production (> 40%), and increases in retirement, re-homing, euthanasia and disposal through Category 2 plants and abattoirs. The absence or banning of horse abattoirs has been shown to have very significant welfare social and economic consequences in the USA. This study described the currently available data on the demographics of unwanted horses in Ireland from 2005 to 2010.

Results

The majority of horses euthanised by practicing veterinarians are destroyed on medical grounds but the number euthanised at the request of welfare groups and the state, as well as welfare related calls and the number of horses involved in these calls and subsequent visits is increasing reflecting the increasing involvement of the veterinary profession in equine welfare. Welfare groups have limited resources and do not have a tradition of recording data, but they too have reported increasing calls, visits and numbers of horses per visit. Welfare groups provide significant service to equine welfare and the community. Local Authorities report similar trends. Over 300 horses were found dead or required immediate or subsequent euthanasia following welfare group and local authority visits in 2010, which is of national concern. The majority of local authority interfaces with unwanted horses are with urban (60%) rather than rural (40%) horses. Mortality figures are poor indicators of non-fatal neglect. More horses were admitted into the care of local authorities than welfare groups, reflecting significant state and taxpayer investment in the control of low value horses. Category 2 plants and abattoirs represent a significant state investment in licensing and control in the national interest. Abattoirs provide an increasingly important and essential service for the disposal of unwanted horses. Despite the increase in unwanted horses, Ireland is a minority contributor to the EU slaughter total.

Conclusions

There is a need for annual demographic data compilation and review of the numbers of unwanted horses and ponies within the horse industry to assist policy makers and legislators.     

Farm management factors associated with bulk tank total bacterial count in Irish dairy herds during 2006/07

Research has shown that total bacterial count (TBC), which is the bacterial growth per ml of milk over a fixed period of time, can be decreased by good hygiene and farm management practices. The objective of the current study was to quantify the associations between herd management factors and bulk tank TBC in Irish spring calving, grass-based dairy herds. The relationship between bulk tank TBC and farm management and infrastructure was examined using data from 400 randomly selected Irish dairy farms where the basal diet was grazed grass. Herd management factors associated with bulk tank TBC were identified using linear models with herd annual total bacterial score (i.e., arithmetic mean of the natural logarithm of bulk tank TBC) included as the dependent variable. All herd management factors were individually analysed in a separate regression model, that included an adjustment for geographical location of the farm. A multiple stepwise regression model was subsequently developed. Median bulk tank TBC for the sample herds was 18,483 cells/ml ranging from 10,441 to 130,458 cells/ml. Results from the multivariate analysis indicated that the following management practices were associated with low TBC; use of heated water in the milking parlour; participation in a milk recording scheme; and tail clipping of cows at a frequency greater than once per year. Increased level of hygiene of the parlour and cubicles were also associated with lower TBC. Herd management factors associated with bulk tank TBC in Irish grazing herds were generally in agreement with most previous studies from confinement systems of milk production.     

Oxidation of glucose, glutamate, and glutamine by isolated ovine enterocytes in vitro is decreased by the presence of other metabolic fuels

The objective of this study was to evaluate oxidative metabolism of glucose, glutamate, and glutamine by isolated ovine enterocytes in the presence of other metabolic fuels in vitro. A mixed mucosal primary cell culture containing enterocytes was isolated from crossbred wether sheep (n = 6) fed a mixed forage-concentrate diet and incubated for 90 min with 1 mM U-14C-glucose, -glutamate, or -glutamine and additional substrates (water as negative control, acetate, propionate, butyrate, glucose, glutamate, or glutamine) at concentrations of 0.1, 1.0, and 10.0 mM. Oxidation of labeled substrates to CO2 and net production of lactate and pyruvate in incubation media were measured. Oxidation of glucose and glutamine to CO2 was decreased (P < 0.05) by 5 to 40% in the presence of additional substrates except acetate. Our observation that glutamine oxidation can be decreased by the presence of additional substrates is contrary to observations in the literature using enterocytes from nonruminants, indicating that ruminant enterocytes might rely on glutamine to a lesser extent as an energy source. Net glucose utilization was decreased (P < 0.05) 16% by propionate (10 mM) compared with control but was not affected by the other additional substrates. Glutamate oxidation to CO2 was decreased 28% (P < 0.05) in the presence of propionate (10 mM) or by 17 and 33% in the presence of glutamine (1.0 and 10 mM, respectively), but not by that of the other additional substrates. Acetate did not affect the oxidation of glucose, glutamate, and glutamine. Propionate decreased (P < 0.05) the oxidation of glucose and glutamate only at the highest concentration (10 mM), indicating that the sparing effects of propionate on substrate oxidation are affected by its concentration in the incubation media. These observations indicate that ruminant enterocytes possess metabolic flexibility for oxidative metabolism of glucose, glutamine, and glutamate depending on the type and concentration of available additional substrates.     

Developmental changes in ketogenic enzyme gene expression during sheep rumen development

Ketogenesis is the conversion of acetyl-CoA to the ketone bodies acetoacetate and beta-hydroxybutyrate (BHBA). In hepatic ketogenesis, which occurs during fasting in both nonruminant and ruminant animals, the source of acetyl-CoA is the mitochondrial oxidation of predominantly long-chain fatty acids. In the mature, fed ruminant animal, the ruminal epithelium is also capable of producing ketone bodies. In this case, the source of acetyl-CoA is the mitochondrial oxidation of butyrate produced by the microbial fermentation of feed. The purposes of this study were to determine ontogenic and dietary effects on ketogenic enzyme gene expression in developing lamb ruminal epithelium. Twenty-seven conventionally reared lambs and twenty-seven milk-fed lambs were slaughtered between 1 and 84 d of age. Six additional milk-fed lambs were weaned (the fed group) or maintained on milk replacer with a volatile fatty acid gavage (the VFA group) until 84 d of age. At slaughter, total RNA was extracted from samples of ruminal epithelium. The expression of the genes encoding acetoacetyl-CoA thiolase, the first enzyme in the ketogenic pathway, and 3-hydroxy-3-methylglutaryl-CoA synthase, the rate-limiting enzyme in the ketogenic pathway in nonruminant liver, were examined. Acetoacetyl-CoA thiolase and 3-hydroxy-3-methylglutaryl-CoA synthase mRNA concentrations increased with age independent of diet. 3-Hydroxy-3-methylglutaryl-CoA synthase mRNA levels in ruminal epithelium obtained from milk-fed lambs were low before 42 d of age, but a marked increase occurred by 42 d of age. At 84 d of age, there were no differences in acetoacetyl-CoA thiolase and 3-hydroxy-3-methylglutaryl-CoA synthase expression due to diet. The pattern of the expression of these genes, in particular, 3-hydroxy-3-methylglutaryl-CoA synthase, parallels the rate of production of BHBA by rumen epithelial cells isolated from the same lambs, which increased to conventionally reared adult levels at 42 d of age and did not differ with diet. In conclusion, development of the ketogenic capacity of the ruminal epithelium occurs as the animal ages, regardless of dietary treatment. Thus, the expression of the genes encoding the ketogenic enzymes are not affected by the presence of VFA in the ruminal lumen.     

Use of computer simulation to teach a systems approach to metabolism

Use of a systems approach, as embodied in the computer simulation model of metabolism of a dairy cow, Molly (Baldwin, 2005), is ideal for teaching nutrition. This approach allows the overall complexity of the comprehensive system to be broken down into smaller manageable subunits that are easier to visualize. Quantitative interactions among nutrients supplied and metabolic production processes can be observed over extended time periods. Using Molly, undergraduate animal science students are able to observe detailed effects of changing dietary inputs, altering genetic milk production potential, and exogenously manipulating metabolism on metabolism of the whole cow. This paper demonstrates how Molly is used in the classroom to teach a systems approach to nutrition using example simulations. Three simulation examples demonstrate exercises examining effects of recombinant bovine somatotropin administration, dietary protein, and amino acid supplementation and nitrogen efficiency on milk production and cow metabolism. These and similar examples have been used to teach nutrition, metabolism, and lactation to undergraduate students for the past 20 yr.     

Textural Images Analysis of Pasta Protein Networks to Determine Influence of Technological Processes

The structure of pasta is largely governed by the presence of a structured protein network. This work analyzed the protein network textures of various cooked pasta products through textural image analysis. Six different pasta types were investigated: reference pasta made from durum semolina; pasta enriched with gluten proteins from soft wheat flour at 10 and 20%; autoclaved pasta; soft wheat flour pasta; and pasta made from reconstituted flour fractions. Pasta samples were sectioned, and each crosssection consisted of three distinct zones (central, intermediate, and external) based on the state of swelling of starch granules for each pasta product. Digital images of the protein network in each zone were acquired using confocal laser scanning microscopy. Textural image analysis was then performed. Similarities and differences in protein network texture were assessed by principal component, stepwise discriminant, and variance analyses. With the exception of autoclaved pasta, protein network structure differed greatly with the position in the pasta. Furthermore the effect of technological treatments was greatly influenced by the position in pasta. The most significant differences in protein network structure were obtained with the autoclaved and 20% protein-enriched samples.