Production of gluten-free wheat starch by peptidase treatment

The potential of peptidase-containing bran extracts from germinated cereals (wheat, emmer, barley) and a peptidase preparation from Aspergillus niger (AN-PEP) to degrade gluten in wheat starch below the threshold for gluten-free foods of 20 mg/kg was compared. The gluten-specific peptidase activity of the peptidases was determined by using gliadin as a protein-based substrate as well as the two celiac-active peptides PQPQLPYPQPQLPY (α-gliadin) and SQQQFPQPQQPFPQQP (γ-hordein). The peptidase activity of AN-PEP exceeded the activities of bran from germinated cereals by a factor up to 690,000. Three wheat starches with initial gluten contents of 110, 1679, and 2070 mg/kg, respectively, were incubated with bran extracts and AN-PEP, lyophilized, and residual gluten was quantitated by a competitive ELISA. Unlike peptidases from bran extracts, AN-PEP was capable of degrading gluten below 20 mg/kg in all starches. The absence of gluten in AN-PEP-treated starches was confirmed by liquid-chromatography-mass spectrometry. The properties of gluten-free starches were comparable to the native starches with the exception of a reduced viscosity after AN-PEP treatment. This problem could be overcome by using higher enzyme concentrations and shorter incubation times or by optimizing AN-PEP production for lower residual α-amylase activity.     

Dogs are more permissive than cats or guinea pigs to experimental infection with a human isolate of Bartonella rochalimae

Bartonella rochalimae was first isolated from the blood of a human who traveled to Peru and was exposed to multiple insect bites. Foxes and dogs are likely natural reservoirs for this bacterium. We report the results of experimental inoculation of two dogs, five cats and six guinea pigs with the only human isolate of this new Bartonella species. Both dogs became bacteremic for 5–7 weeks, with a peak of 10³–10⁴ colony forming units (CFU)/mL blood. Three cats had low bacteremia levels (< 200 CFU/mL) of 6–8 weeks’ duration. One cat that remained seronegative had two bacterial colonies isolated at a single culture time point. A fifth cat never became bacteremic, but seroconverted. None of the guinea pigs became bacteremic, but five seroconverted. These results suggest that dogs could be a reservoir of this strain of B. rochalimae, in contrast to cats and guinea pigs.     

Vegetation Production Responses to October Grazing in the Nebraska Sandhills

Understanding the long-term effect of summer grazing date and fall stocking rate on herbage production is critical to extending the grazing season in the Nebraska Sandhills. A study was conducted from 1997 to 2002 at the Gudmundsen Sandhills Laboratory located near Whitman, Nebraska, to determine the herbage production response to summer grazing date and October stocking rate on two different sites. Site 1 was dominated by warm-season grasses and site 2 was dominated by cool-season graminoids. At each site, three 0.37-ha pastures were constructed in each of four blocks before application of summer grazing treatments. Pastures in each block were grazed at 0.5 animal-unit months (AUM) · ha^?1 in June or July, or were deferred from summer grazing. Following summer grazing treatments, October stocking rate treatments (no grazing or 1.0, 2.0, or 3.0 AUM · ha^?1) were applied to subunits of each summer grazing date pasture during mid-October. Vegetation was sampled in each pasture in mid-June and mid-August and sorted by functional group to determine the effect of 5 yr of grazing treatments on herbage production and residual herbage. Herbage production was not affected by summer or October grazing treatments on the warm-season grass–dominated site. Increasing October stocking rate, however, reduced cool-season graminoid production and subsequent herbage production 25% by year 5 of the study. Residual herbage at both sites at the end of the October grazing periods explained as much as 16% to 34% of subsequent year’s herbage production. Grazing managers in the Nebraska Sandhills can extend the grazing season by lightly stocking pastures in the summer to facilitate additional fall grazing. Heavy stocking in October over several years on cool-season–, but not warm-season–, dominated sites will reduce production of cool-season graminoids on these sites.     

Late nitrogen application increased protein concentration but not baking quality of wheat

Late application of nitrogen (N) fertilizers at heading or anthesis is usually performed to produce wheat (Triticum aestivum L.) with high bread‐making quality. However, increasing energy costs and ecological problems due to N losses call for efficient and simplified N fertilization strategies. This study aimed to investigate the effect of late N fertilization on grain protein quality and thus baking quality and to evaluate if similar wheat quality can be maintained without late N application. Field experiments with two winter wheat cultivars differing in quality groups were conducted. The fertilization treatments comprised a rate of 220 kg N ha^?1 applied in two or three doses (referred to as split N application), and 260 kg N ha^?1 applied in four doses (additional late N fertilization) with different N fertilizer types. The results show that although split N application had no effect on grain protein concentration (GPC), it affected N partitioning in the grain, increasing mainly the concentration and proportion of the glutenin fraction. As a result, baking quality was improved by split N application. Late N fertilization enhanced GPC and the relative abundance of certain high molecular weight glutenin subunits (HMW‐GS). However, it had no effect on N partitioning in the grain and did not further improve baking quality. No obvious differences were found between N fertilizer types on grain yield and quality. The N fertilization effect was more pronounced on the wheat cultivar whose baking quality was more dependent on protein concentration. In evaluating baking quality of wheat flour, gliadin and glutenin proportions were better correlated with loaf volume than the overall protein concentration.     

Concentrations of low and high molecular weight thiols in wheat dough as affected by different concentrations of ascorbic acid

Different amounts of ascorbic acid (AA) were added to flour, and the concentrations of low and high molecular weight thiols in the dough were determined. For the determination of the low molecular weight thiols, glutathione, cysteine, and the corresponding disulfides, an isotope dilution assay with a (14)C-labeled internal standard was used. For the determination of the high molecular weight thiols, a method was developed that involved derivatization of dough with Ellman's reagent, removal of excess reagent by dialysis, micro-Osborne fractionation, release of the label by reduction, and determination of reduced Ellman's reagent by reversed-phase high-performance liquid chromatography. Mixing of flour without AA led to a decrease of the glutathione and an increase of the cysteine concentration. Addition of AA reduced the concentration of both thiols to a minimum when 125 mg of AA/kg of flour was applied. Furthermore, the concentrations of high molecular weight thiols in the glutenins of flours from different wheat cultivars were determined. The values ranged from 5.6 to 8.2 micromol/kg of protein and showed a correlation between flour quality and SH concentration. On addition of AA and mixing of a dough, the concentrations of the protein thiols in the glutenins isolated from the dough increased to a maximum when 100 mg of AA/kg of flour was added. Higher concentrations of AA led to a decrease of the SH concentration. The last results are not in accordance with previously published data or with current hypotheses about the mechanism of the AA improver action.     

Feline infectious peritonitis presenting as a tumour in the abdominal cavity

This paper describes six cases of feline infectious peritonitis (FIP) in which an abdominal tumour had been suspected clinically. Pathological changes were mainly restricted to the massive enlargement of a mesenteric lymph node due to necrogranulomatous lymphadenitis. FIP was diagnosed on the basis of the immunohistological demonstration of coronavirus antigen in intact macrophages within the necrogranulomatous lesions. In the affected lymph node lymphoid tissue was either almost completely effaced or restricted to follicles composed mainly of variable numbers of blasts. From one to many plasma cells positive for coronavirus-specific antibodies were present in the marginal sinuses or capsules. In addition, necrogranulomas were present in the gut-associated lymphoid tissue of the caecum of one cat, and adjacent to the affected lymph node of another.     

Association between patient-related factors and risk of calcium oxalate and magnesium ammonium phosphate urolithiasis in cats

OBJECTIVE: To determine whether breed, age, sex, or reproductive status (i.e., neutered versus sexually intact) was associated with the apparent increase in prevalence of calcium oxalate (CaOx) uroliths and the decrease in prevalence of magnesium ammonium phosphate (MAP) uroliths in cats over time. DESIGN: Case-control study. ANIMALS: Case cats consisted of cats with CaOx (n = 7,895) or MAP (7,334) uroliths evaluated at the Minnesota Urolith Center between 1981 and 1997. Control cats consisted of cats without urinary tract disease admitted to veterinary teaching hospitals in the United States and Canada during the same period (150,482). PROCEDURE: Univariate and multivariate logistic regression were performed. RESULTS: British Shorthair, Exotic Shorthair, Foreign Shorthair, Havana Brown, Himalayan, Persian, Ragdoll, and Scottish Fold cats had an increased risk of developing CaOx uroliths, as did male cats and neutered cats. Chartreux, domestic shorthair, Foreign Shorthair, Himalayan, Oriental Shorthair, and Ragdoll cats had an increased risk of developing MAP uroliths, as did female cats and neutered cats. Cats with CaOx uroliths were significantly older than cats with MAP uroliths. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that changes in breed, age, sex, or reproductive status did not contribute to the apparent reciprocal relationship between prevalences of CaOx and MAP uroliths in cats during a 17-year period. However, cats of particular breeds, ages, sex, and reproductive status had an increased risk of developing CaOx and MAP uroliths.