Lack of toxicity of a non-sporidesmin-producing strain of Pithomyces chartarum in cell culture and when dosed to lambs

In New Zealand the fungus Pithomyces charturum normally produces sporidesmin, a mycotoxin, which is responsible for the hepatogenous photosensitisation disease known as facial eczema. Cultures from an isolate of P. charturum, which does not produce sporidesmin, were examined by cell culture and by dosing to lambs to determine whether other toxic metabolites were produced. Acute and long term toxicity studies were conducted with the toxic response being assessed by weight changes, postmortem and histological examination of tissues, blood biochemistry and haematology tests. An extract from a sporidesmin-producing isolate was highly toxic in cell culture, while extracts of the nonsporidesmin-producing isolate did not cause a cytotoxic response to HEp 2 cells. After dosing with a sporidesmin-producing isolate, lambs developed liver lesions and clinical signs of facial eczema. Serum biochemistry changes occurred which were consistent with sporidesmin poisoning. Lambs dosed with the nonsporidesmin-producing isolate, at the rate of thirty times the number of spores of the sporidesmin-producing isolate, showed no observable toxic effects. All organs were of normal appearance, and histological examination of tissues, blood biochemistry and haematology results showed no abnormal changes. Similarly, long term dosing of extracts of the nonsporidesmin-producing isolate, at a rate equivalent to 100,000 spores/g of grass, produced no indication of a toxic response. It was concluded that the nonsporidesmin-producing isolate of P. churtarum contained no toxic metabolites in significant concentration.     

Competition of a sporidesmin-producing Pithomyces strain with a non-toxigenic Pithomyces strain

Sporidesmin, a mycotoxin produced by some strains of Pithomyces chartarum, is responsible for the hepatogenous photosensitisation disease facial eczema, which causes severe losses in agricultural revenue in New Zealand. A sporidesmin-producing strain of P. chartarum, isolated in New Zealand, was grown in co-culture with a South African strain that does not produce the mycotoxin. Competition occurred between the two strains when grown both on agar plates and on dried ryegrass, with a significant decrease in the total amount of sporidesmin produced. Biological control of toxigenic P. chartarum can thus occur under laboratory conditions, raising the possibility of similar control in the field situation.     

大肠杆菌0157的分离鉴定

0157为大肠杆菌的一种血清型,该血清型于1982年首次被鉴定为人发生该病的病因[1],在此之前这种血清型很少分离到.这种血清型归类为肠出血性大肠杆菌(EHEC),可以引起血痢和产生强毒素.据有关文献介绍,在采集的牛肉、猪肉和禽肉产品检测后曾有0157大肠杆菌的阳性报道.     

Release of deuterated nonenal during beer aging from labeled precursors synthesized in the boiling kettle.

The use of labeled nonenal enabled the demonstration that the appearance of the cardboard flavor in finished beer comes from lipid auto-oxidation during wort boiling and not from lipoxygenasic activity during mashing. Free trans-2-nonenal produced by linoleic acid auto-oxidation in the kettle disappears, owing to retention by wort amino acids and proteins. This binding linkage protects trans-2-nonenal from yeast reduction but is reversible, allowing release of the compound at lower pH during aging. Labeled trans-2-nonenal is detected after aging when deuterated precursors form in the boiling kettle. The amount of alkenal released correlates with the concentration of reversible associations in the pitching wort. This work brings new illumination to the formation of trans-2-nonenal and overturns many previous hypotheses. It also explains why a reduction in the beer pH intensifies the cardboard flavor.     

Monoclonal antibodies as probes for the changes in antigenicity of bovine and porcine aspartyl proteases with pH.

Two monoclonal antibodies, 918(4) and 139(7), directed against either bovine or porcine pepsin, respectively, were retained among 365 positive hybridoma clones. These monoclonal antibodies were characterized by using both indirect and sandwich ELISA. Characterization of these monoclonal antibodies was further performed by the biospecific interaction analysis (BIA-core analysis). Then, they were used as antigenic probes to study the changes in antigenicity of both bovine and porcine pepsins induced by pH. The results demonstrated the importance of the conformational change in both catalytic activities and antigenic determinant accessibility of bovine and porcine pepsins. Furthermore, our results suggest that changes in the conformation due to pH can be detected by specific monoclonal antibodies.     

Glycerol,Methyl‐Formamide and Dimethyl‐Formamide in Canine Semen Cryopreservation

The aim of the present work was to compare the efficiency of methyl‐formamide (MF), dimethyl‐formamide (DF) and glycerol (GL) as cryoprotectants in canine semen cryopreservation. For the experiment, pooled semen was submitted to one of the three cryoprotectants, with a final concentration of 3% in egg yolk–TRIS extender. Semen was subjectively evaluated for total and progressive motility, vigour and morphology. Sperm membrane functional integrity was assessed by hypo‐osmotic swelling test (HOST), and longevity was assessed using the thermoresistance test (TRT). Fresh semen showed normal physical and morphological characteristics. After thawing, differences were observed between semen frozen using GL and DF, regarding total and progressive motility and vigour (p < 0.05), but not between MF and GL or MF and DF. Means for total motility, progressive motility, vigour and morphologically normal spermatozoa were, respectively, 69.0 ± 5.4%, 61.0 ± 7.4%, 2.9 ± 0.5 and 57.1 ± 5.0% for GL; 59.0 ± 8.9%, 50.0 ± 10.0%, 2.5 ± 0.7 and 66.9 ± 7.7% for MF; and 44.0 ± 21.0%, 37.0 ± 19.8%, 2.1 ± 0.6 and 61.1 ± 5.5% for DF. On HOST, GL was superior (p < 0.05) to MF and DF (57.8 ± 12.4%, 35.8 ± 18.4% and 34.4 ± 9.4%, respectively). During the TRT, both GL and MF were superior to DF, with no differences between GL and MF. In conclusion, the use of MF as cryoprotectant showed results similar to GL, and can be considered as an alternative in canine semen cryopreservation. Further studies testing different concentrations of MF may improve its effects on cryopreservation of canine semen.     

The development and integrity of equine pre‐antral follicles cultured in vitro with follicle‐stimulating hormone (FSH) supplementation

This study investigated the effects of different concentrations of FSH (10, 50, 100 and 200 ng/ml) in supplemented MEM+ on the development of equine pre‐antral follicles that were cultured in vitro for 2 or 6 days. The ovaries (n = 5) from mares in seasonal anoestrus were collected from a local abattoir. Ten ovarian tissue fragments of approximately 3 × 3 × 1 mm were obtained from each animal. The fragments were cultured in situ for 2 days (D2) or 6 days (D6) in MEM+ or MEM+ supplemented with FSH at four different concentrations, establishing the following 11 groups: control (D0); MEM + (D2); MEM + (D6); MEM + 10 ng/ml of FSH (D2); MEM + 10 ng/ml of FSH (D6); MEM + 50 ng/ml of FSH (D2); MEM + 50 ng/ml of FSH (D6); MEM + 100 ng/ml of FSH (D2); MEM + 100 ng/ml of FSH (D6); MEM + 200 ng/ml of FSH (D2); and MEM + 200 ng/ml of FSH (D6). Follicles were observed in only 9.65% (388 of 4,018) of the histological sections. Of the 861 follicles evaluated, 488 were in the primordial stage, and 373 were in various developmental stages; 59.7% were morphologically normal. Regarding the integrity of the pre‐antral follicles, the groups with 100 ng/ml FSH of 2‐days culture as well as 50, 100 and 200 ng/ml FSH of 6‐days culture provided the best results. In conclusion, the in vitro culture of abattoir‐derived equine ovarian fragments presented better morphological integrity when supplemented with FSH for 6 days, in comparison with the MEM culture group. However, no clear effects were observed with FSH regarding the promotion of activation from a primordial to a developing follicle.     

Effect of feeding flax or linseed meal on progesterone clearance rate in ovariectomized ewes

Ovariectomized ewes (n=22; 68.76+/-2.34 kg initial body weight; 2.9+/-0.1 initial body condition score) were individually fed one of three diets: (1) control (phytoestrogen-free; n=7), (2) flax containing diet (n=8), or (3) linseed meal (LSM) containing diet (n=7) to investigate the rate of progesterone (P4) clearance. On day 20 of feeding (day 0=initiation of treatment), a P4 releasing device (CIDR) was placed in the vagina and jugular blood samples were obtained prior to CIDR insertion and 15, 30, 60, and 120 min following CIDR insertion. Further, blood samples were obtained daily between days 21 and 24. On day 25, blood samples were retrieved prior to CIDR removal and 2, 5, 10, 15, 30, 60, 120, and 360 min following CIDR removal. There was no difference in initial or final body weight or body condition score and there were no time by diet interactions on P4 clearance. The fractional rate of P4 uptake measured prior to CIDR insertion through day 4 following insertion tended to be greater (P=0.07) in LSM fed ewes (508.75+/-71.37%/min) compared to flax (295.39+/-66.76%/min) and control fed (287.54+/-71.37%/min) ewes. Diet tended (P=0.10) to influence P4 clearance rate when measured from prior to CIDR removal through 120 min following CIDR removal with LSM fed ewes having a greater (1.26+/-0.2) fractional rate constant than flax (0.929+/-0.09) and control fed (0.922+/-0.09) ewes. Flax fed ewes also had more (P<0.01) omega-3 fatty acids and total fatty acids in plasma. Reports of increased pregnancy rates in dairy cows fed flax may relate to P4 metabolism.     

Genomic constitution of a partial amphiploid OK7211542 used as a source of immunity to barley yellow dwarf virus for bread wheat

A combination of genomic in situ hybridization (GISH) and meiotic pairing analysis of crosses between a series of 2n= 56 partial amphiploids confirmed that the alien genome of the BYDV-immune Agro-tricum line OK7211542 is derived from Thinopyrum ponticum and not from Thinopyrum intermedium. The evidence from meiotic pairing analysis indicated that the chromosome constitution of OK7211542 is similar to another Agrotricum line, ORRPX, which was derived from a cross of wheat and Th. ponticum, but different from other Agrotricum lines, Zhong 5 and TAF 46 which were derived from the crosses between wheat and Th. intermedium. The GISH analysis confirmed that OK7211542 contained one complete set of 14 Th. ponticum chromosomes, in which no S chromosome was present in the alien genome. GISH also detected a small alien translocation attached to one of the wheat chromosomes, a result that was consistent with the pairing data.     

Hazard Analysis and Critical Control Points System for a Bull Semen Production Centre

Bull semen production centres (SPC) generally present satisfactory quality control for sperm processing, but non‐standardized hygiene procedures. This study describes a Hazard Analysis and Critical Control Points (HACCP) system developed for bull SPC and subsequently implemented in a commercial SPC. After the identification of hazards at each step of semen processing and the determination of their risk and severity, monitoring and corrective procedures were designed to assess the system's efficiency. The HACCP system identified six microbiological hazards, 10 physical hazards, four chemical hazards and three critical control points. After the establishment of Good Processing Practices, Standard Operating Procedures and Standard Sanitizing Operating Procedures, the system was validated through an audit, to identify eventual failures and to define measures to correct them.