Modeling First Break Milling of Debranned Wheat Using the Double Normalized Kumaraswamy Breakage Function

Debranning of wheat affects flour quality, initially by altering the breakage of wheat kernels during first break. The double normalized Kumaraswamy breakage function was applied to model the effect of debranning on wheat breakage during first break milling, in which type 1 breakage describes a relatively narrow distribution of midsized particles, whereas type 2 breakage describes a wide size range of predominantly small particles extending to very large particles. Mallacca (hard) and Consort (soft) wheats were debranned and milled at three roll gaps under sharp‐to‐sharp and dull‐to‐dull dispositions. Type 1 breakage increased at longer debranning times, whereas type 2 breakage decreased, for both wheat varieties under both dispositions. Sharp‐to‐sharp milling tended to produce more type 1 breakage than dull‐to‐dull. A mechanism of wheat breakage is proposed to explain the coproduction of very large and small particles via type 2 breakage and, hence, the effect of debranning. The proposed mechanism is that small particles of endosperm arise from scraping of large flat particles of wheat bran under the differential action of the rolls, such that removal of the bran reduces the production of the large bran particles and thus reduces the opportunity for the scraping mechanism that produces the very small particles.     

Bran Characteristics and Bread‐Baking Quality of Whole Grain Wheat Flour

Variations in physical and compositional bran characteristics among different sources and classes of wheat and their association with bread‐baking quality of whole grain wheat flour (WWF) were investigated with bran obtained from Quadrumat milling of 12 U.S. wheat varieties and Bühler milling of six Korean wheat varieties. Bran was characterized for composition including protein, fat, ash, dietary fiber, phenolics, and phytate. U.S. soft and club wheat brans were lower in insoluble dietary fiber (IDF) and phytate content (40.7–44.7% and 10.3–17.1 mg of phytate/g of bran, respectively) compared with U.S. hard wheat bran (46.0–51.3% and 16.5–22.2 mg of phytate/g of bran, respectively). Bran of various wheat varieties was blended with a hard red spring wheat flour at a ratio of 1:4 to prepare WWFs for determination of dough properties and bread‐baking quality. WWFs with U.S. hard wheat bran generally exhibited higher dough water absorption and longer dough mixing time, and they produced smaller loaf volume of bread than WWFs of U.S. soft and club wheat bran. WWFs of two U.S. hard wheat varieties (ID3735 and Scarlet) produced much smaller loaves of bread (<573 mL) than those of other U.S. hard wheat varieties (>625 mL). IDF content, phytate content, and water retention capacity of bran exhibited significant relationships with loaf volume of WWF bread, whereas no relationship was observed between protein content of bran and loaf volume of bread. It appears that U.S. soft and club wheat bran, probably owing to relatively low IDF and phytate contents, has smaller negative effects on mixing properties of WWF dough and loaf volume of bread than U.S. hard wheat bran.     

Zinc Blotting Assay for Detection of Zinc‐Binding Prolamin in Barley (Hordeum vulgare) Grain

In plants, zinc is commonly found bound to proteins. In barley (Hordeum vulgare), major storage proteins are alcohol‐soluble prolamins known as hordeins, and some of them have the potential to bind or store zinc. ⁶⁵Zn overlay and blotting techniques have been widely used for detecting zinc‐binding protein. However, to our knowledge so far this zinc blotting assay has never been applied to detect a prolamin fraction in barley grains. A radioactive zinc (⁶⁵ZnCl₂) blotting technique was optimized to detect zinc‐binding prolamins, followed by development of an easy‐to‐follow nonradioactive colorimetric zinc blotting method with a zinc‐sensing dye, dithizone. Hordeins were extracted from mature barley grain, separated by SDS‐PAGE, blotted on a membrane, renatured, overlaid, and probed with zinc; subsequently, zinc‐binding specificity of certain proteins was detected either by autoradiography or color formation. The dithizone staining method gave similar reproducibility to the radioactive blotting. The detected zinc‐binding protein was identified as B‐hordein by Western blotting.     

Expression,Purification, and Functional Analysis of Three Low‐Molecular‐Weight Glutenin Subunits from Wheat Cultivar Cheyenne

Glutenins, which form the network of gluten protein, are of great importance for the quality of flour products. Glutenins can be divided into HMW and LMW subunits according to molecular weight. Three genes for LMW glutenin subunits (LMW‐GS), named lmw‐cnd1, lmw‐cnd2, and lmw‐cnd3 with open reading frames of 1,053, 903, and 969 bp, respectively, were cloned from wheat cultivar Cheyenne. Heterologous expression vectors of the three LMW‐GS were constructed, and the recombinant proteins LMW‐CND1, LMW‐CND2, and LMW‐CND3 were overexpressed in Escherichia coli. After cell disruption with ultrasound, target proteins of high purity were obtained by using Ni²⁺ affinity chromatography. Farinograph and TAPlus measurements were used to investigate the effects of the three LMW‐GS on the characteristics of flour and dough. The results showed that the addition of each LMW‐GS can lead to an increase in the elasticity of the dough. Moreover, LMW‐CND2 and LMW‐CND3 promoted the strength of the dough. All three LMW‐GS caused a decrease of hardness and increase of springiness and cohesiveness of dough according to texture profiling results. Consequently, all three LMW‐GS have positive effects on the processing characteristics of dough and can improve bread quality to different extents.     

Foetal Protection against Bovine Virus Diarrhoea Virus after Two‐step Vaccination

In order to assess the efficacy of a two‐step vaccination protocol with respect to foetal protection against transplacental infections with bovine virus diarrhoea virus (BVDV) with special attention to BVDV‐2 seronegative heifers were vaccinated with an inactivated BVDV‐1 vaccine and boostered with a modified live BVDV‐1 vaccine after 4 weeks. A second group was left unvaccinated as control. Between days 30 and 120 of pregnancy the heifers of both groups were intranasally challenged with a mixture of BVDV‐1 and ‐2. All heifers of the vaccinated group gave birth to nine clinically healthy, seronegative (precolostral) and BVDV‐free calves. In contrast in the control group four BVDV viraemic underdeveloped calves were born. Additionally, one calf was stillborn and another viraemic calf was not viable and died 2 days after birth. All six calves of the control group were viraemic with BVDV‐2. This study demonstrated for the first time that two‐step vaccination of breeding cattle with a modified live BVDV vaccine 4 weeks after application of an inactivated BVDV vaccine was capable of providing a foetal protection against transplacental infection with BVDV‐2.     

Nuclear transfer using a bovine mammary epithelial cell line (BMEC)

The developmental potential of nuclei from a bovine mammary epithelial cell line (BMEC) in nuclear transfer was investigated. For nuclear transfer donors, BMEC cells (passage 15) were cultured for 4–5 days after seeding at cell densities of 1.0 × 10⁵ cells/cm² (high‐density group) or 0.8 × 10⁴ cells/cm² (low‐density group). First, the effective electric stimulation for fusion of enucleated oocytes with BMEC cells was examined. Fusion rates reached maximum with two DC pulses of 30 V/150 µm for 20 µs in the high‐density group and with two DC pulses of 25 V/150 µm for 10 µs in the low‐density group. The fusion rate (37.5%) in the high‐density group was significantly (P < 0.005) lower than in the low‐density group (71.4%). Second, the in vitro developmental potential of nuclear transfer embryos derived from BMEC cells was examined. In the high‐density and low‐density groups, 18.8% and 24.1% of fused oocytes, respectively, developed to blastocyst stage. The results of this study indicate that nuclei from BMEC cells support the development of nuclear transfer embryos to the blastocyst stage and that the efficiency of oocyte–cell fusion is affected by the culture conditions of the donor BEMC cells before nuclear transfer.     

Electron microscopic observation of the respiratory tract of SPF piglets inoculated with Mycoplasma hyopneumoniae.

Seven hysterectomy derived piglets were repeatedly challenged with Mycoplasma hyoneumoniae during the first week of life. Samples of trachea, bronchi and lung tissue collected 2–11 weeks post-inoculation (p.i.) were examined using light and electron microscopy. Autoradiography was used to study in more detail the site of M. hyopneumoniae multiplication. Gross lesions were observed in lung tissue and were characterized by hyperplasia of the epithelium and an increased mononuclear cell accumulation in perivascular and peribronchiolar areas. Mild lesions of the trachea and the bronchi, including epithelial hyperplasia and infiltration of the lamina propria by inflammatory cells, were noted.

Electron microscopy showed that, 2–6 weeks p.i., changes in the mid-trachea and bronchi surface consisted of the loss of cilia. Mycoplasmas covered tufts of cilia remaining on the epithelial cell surface. Scanning and transmission electron micrographs showed that they were predominantly found closely associated with the top of cilia. No specialized terminal structure could be seen and no mycoplasma cells were identified lying free in the lumen nor in close contact with the plasma membrane of cells of microvilli. Some fine fibrils radiating from one mycoplasma to another or to cilia were seen at higher magnification by scanning electron microscopy. Six to eleven weeks p.i., a disrupted epithelial surface lacking cilia was observed. Cells were desquamated and shed into the lumen with cellular remains containing droplets of mucus.

Autoradiography revealed that label corresponded to the observed mycoplasma distribution. At the top of cilia, a high density of labeling was visible in the zone of high mycoplasma concentration. Therefore, incorporation of the label in the mycoplasma is proof or their multiplication in the trachea.

The intimate association between the mycoplasma and cilia may be an important factor in the pathogenesis of the disease caused by M. hyopneumoniae (swine enzootic pneumonia).     

The Effect of Pasteurella haemolytica A2 Infection on Phagocytosis Efficiency of Caprine Broncho‐Alveolar Macrophages

An experiment was designed to study the in vivo effect of Pasteurella haemolytica A2 infection on the phagocytosis activity of caprine broncho‐alveolar macrophages and the extent of pneumonic lesions. Twelve healthy local Kacang goats, about 7 months of age, were divided into two groups of six. Goats in group 1 were inoculated intratracheally with 4 ml inoculum containing 2.8 × 10⁹ colony‐forming units (CFU)/ml of Staphylococcus aureus. Goats in group 2 were inoculated intratracheally with 4 ml of inoculum containing 9.5 × 10⁸ CFU/ml of Pasteurella haemolytica A2 isolated earlier from pneumonic lungs of goat. At intervals of 3 and 7 days post‐challenge five goats from each group were killed and the lungs were washed with sterile phosphate‐buffered saline. Smears were prepared from the lung washing fluid and the number of macrophages with phagocytic activity was determined. At day 3 post‐infection, goats of both groups showed a similar pattern of pneumonic lesion. The lung washing fluid of goats in group 2 was found to contain numerous neutrophils and macrophages. Goats in group 2 showed significantly (P < 0.05) higher extent of lung lesions than group 1. Similarly, the average extent of lung lesions was significantly (P < 0.05) more severe in group 2 at day 7 post‐infection. The lung washing fluid contained mostly macrophages. The phagocytic activity following S. aureus infection was more efficient and significantly (P < 0.01) higher compared with infection by P. haemolytica A2. There were weak correlations between the extent of pneumonic lesion and the phagocytic activity. Thus, goats with poor phagocytic activity were likely to develop more extensive lung lesions.     

Identification of an Intrinsic Platelet Function Defect in Spitz Dogs

A recently identified intrinsic platelet function defect in 2 Spitz dogs is described. Both affected dogs had a history of chronic intermittent bleeding primarily from the nasal, oral, and gastrointestinal mucosa. Platelet aggregation in response to adenosine diphosphate (ADP), collagen, and platelet activating factor (PAF) was absent; however, platelet shape change did occur. Platelets aggregated in response to gamma thrombin, although a delayed onset and a reduced velocity of aggregation were present. Platelet ¹⁴C-serotonin release was diminished in response to collagen and PAF. Glycoprotein Illa was detected on the surface of platelets by flow cytometry. Platelets were morphologically normal under light and electron microscopy. Two male Spitz dogs, related to one of the affected dogs, did not have a bleeding diathesis. Collagen-induced platelet aggregation, however, was diminished in these 2 dogs. This platelet defect most closely resembles the defect described in Basset hounds.