Isolation and molecular characterisation of Neospora caninum in cattle in New Zealand     

CM Okeoma  NB Williamson  WE Pomroy  KM Stowell  LM Gillespie 《New Zealand veterinary journal》2013,61(6):364-370

AIM: To isolate Neospora caninum from the brains of naturally infected cattle and use molecular techniques to characterise the isolates.

METHODS: Neospora caninum tachyzoites were isolated in Vero cell culture from the brains of a cow and two calves. The isolates were characterised using polymerase chain reaction (PCR) methods, DNA sequencing, an immunofluorescent anti-body test (IFAT), transmission electron microscopy (TEM), and immunohistochemistry (IHC). The brains of the three cattle were subjected to histopathological examination. A pathogenicity study was conducted in 120 BALB/c mice.

RESULTS: Neospora caninum tachyzoites were isolated from all three cases and first observed in vitro between 14 and 17 days post-inoculation. Parasites were sub-cultured and maintained in Vero cell culture for more than 6 months. PCR products were generated for all three isolates, using two different primers. Sequencing of the PCR products and a subsequent BLAST search identified the isolates as N. caninum. In addition, the isolates tested positive using IFAT and IHC, and ultrastructure revealed by TEM was characteristic of N. caninum. Histopathological examination revealed lesions characteristic of N. caninum in 1/3 brains. In the pathogenicity study using BALB/c mice, the mortality rate was 3–7%.

CONCLUSION: This was the first successful isolation of N. caninum in New Zealand confirmed using molecular characterisation tests.  相似文献   

Lectin Histochemistry as a Tool to Identify Apoptotic Cells in the Seminiferous Epithelium of Syrian Hamster (Mesocricetus auratus) Subjected to Short Photoperiod     

V Seco‐Rovira  E Beltrán‐Frutos  C Ferrer  MM Sánchez‐Huertas  JF Madrid  FJ Saez  LM Pastor 《Reproduction in domestic animals》2013,48(6):974-983

Lectins have been widely used to study the pattern of cellular glycoconjugates in numerous species. In the process of cellular apoptosis, it has been observed that changes occur in the membrane sugar sequences of these apoptotic cells. The aim of our work was to identify which lectins, out of an extensive battery of the same (PNA, SBA, HPA, LTA, Con‐A, UEA‐I, WGA, DBA, MAA, GNA, AAA, SNA), show affinity for germinal cells in apoptosis, at what stage of cell death they do so and in which germinal cell types they can be detected. For this, we studied testis sections during testicular regression in Syrian hamster (Mesocricetus auratus) subjected to short photoperiod. Several lectins showed an affinity for the glycoconjugate residues of germ cells in apoptosis: Gal β1,3‐GalNAcα1, α‐d ‐mannose, N‐acetylgalactosamine and l ‐fucose. Furthermore, lectin specificity was observed for some specific germinal cells and in certain stages of apoptosis. It was also observed that one of these lectins (PNA) showed affinity for Sertoli cells undergoing apoptosis. Therefore, we conclude that the use of lectin histochemistry could be a very useful tool for studying apoptosis in the seminiferous epithelium because of the specificity shown towards germinal cells in pathological or experimentally induced epithelial depletion models.  相似文献   

Extraction of β‐Glucan from Oats for Soluble Dietary Fiber Quality Analysis     

Douglas C. Doehlert  Senay Simsek  Michael S. McMullen 《Cereal Chemistry》2012,89(5):230-236

Extraction protocols for β‐glucan from oat flour were tested to determine optimal conditions for β‐glucan quality testing, which included extractability and molecular weight. We found mass yields of β‐glucan were constant at all temperatures, pH values, and flour‐to‐water ratios, as long as sufficient time and enough repeat extractions were performed and no hydrolytic enzymes were present. Extracts contained about 30–60% β‐glucan, with lower proportions associated with higher extraction temperatures in which more starch and protein were extracted. All commercial starch hydrolytic enzymes tested, even those that are considered homogenous, degraded β‐glucan apparent molecular weight as evaluated by size‐exclusion chromatography. Higher concentration β‐glucan solutions could be prepared by controlling the flour‐to‐water ratio in extractions. Eight grams of flour per 50 mL of water generated the highest native β‐glucan concentrations. Routine extractions contained 2 g of enzyme‐inactivated flour in 50 mL of water with 5mM sodium azide (as an antimicrobial), which were stirred overnight, centrifuged, and the supernatant boiled for 10 min. The polymer extracted had a molecular weight of about 2 million and was stable at room temperature for at least a month.  相似文献   

Report of the EPPO Workshop on setting Ct cut‐off values for real‐time PCR     

M. McMullen  F. Petter 《EPPO Bulletin》2014,44(2):165-165

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Starch Characteristics of Waxy and Nonwaxy Tetraploid (Triticum turgidum L. var. durum) Wheats     

L. A. Grant  N. Vignaux  D. C. Doehlert  M. S. McMullen  E. M. Elias  S. Kianian 《Cereal Chemistry》2001,78(5):590-595

Manufacture of pasta products is paramount for durum wheat (Triticum turgidum L. var. durum). The recent development of waxy durum wheat containing starch with essentially 100% amylopectin may provide new food processing applications and present opportunities for value‐added crop production. This investigation was conducted to determine differences in some chemical and functional properties of waxy durum starch. Starch was isolated from two waxy endosperm lines and four nonwaxy cultivars of durum wheat. One of the waxy lines (WX‐1) was a full waxy durum wheat whereas the other line (WX‐0) was heterogeneous, producing both waxy and nonwaxy seed. Effects on starch swelling, solubility, pasting, gelatinization, and retrogradation were examined. The full waxy starch had four times more swelling power than the nonwaxy durum starches at 95°C, and was also more soluble at three of the four temperatures used. Starch pasting occurred earlier and peak viscosities were greater for starches from both waxy lines than for the nonwaxy starches, but their slurries were less stable with continued stirring and heating. Greater energy was required to melt gelatinized waxy starch gels, but no differences were found in either refrigerated storage or freeze‐thaw retrogradation, as determined by differential scanning calorimetry. The results of this investigation showed some significant differences in the starch properties of the waxy durum wheat lines compared to the nonwaxy durum wheats.  相似文献   

Optimizing Conditions for Experimental Oat Dehulling     

Douglas C. Doehlert  Michael S. McMullen 《Cereal Chemistry》2001,78(6):675-679

The determination of groat percentage in experimental oat breeding lines requires the dehulling of oats. Here, we report the results of our efforts to optimize dehulling conditions so that the most accurate and reliable result can be obtained. Hand dehulling was always reliable and accurate, however, it was the most time‐consuming and tedious of the methods studied. Two mechanical methods of oat dehulling, compressed‐air dehulling and impact dehulling, also frequently provided reliable results, however, results were strongly influenced by dehulling conditions. Optimal dehulling conditions represented compromises between unfavorable extremes. Correct aspiration strength was critical to accurate groat percentage determination. We have found that a secondary aspiration is highly desirable after compressed‐air dehulling to remove hulls remaining with the groats after dehulling. Also, increased mechanical stress on oats as exerted either by the number of passes through the impact dehuller, or by the air pressure in the compressed‐air dehuller, resulted in higher dehulling efficiency, but increased groat breakage as well. Dehulling efficiency decreased as moisture increased from 7.5 to 15%, but increased as moisture was further increased to 30%. In contrast, groat breakage with impact dehulling decreased as moisture increased from 7.5 to 30%. A new equation for groat percentage calculation has been introduced where the mass of hulled oats remaining after dehulling is subtracted from the mass of the original oat sample, so that poor dehulling efficiency does not influence the groat percentage.  相似文献   

Vetch toxicosis in cattle grazing Vicia villosa ssp dasycarpa and V benghalensis     

PAW HARPER  RW COOK†  PA GILL†  GC FRASER†  LM BADCOE‡  JM POWER§ 《Australian veterinary journal》1993,70(4):140-144

SUMMARY The epidemiological, clinical and pathological features of a disease syndrome in adult cattle grazing woolly-pod vetch (Vicia villosa ssp dasycarpa) or popany vetch (V benghalensis) are reported. Outbreaks of toxlcosis occurred between midwinter and midsummer in 3 dairy and 6 beef herds on the north coast of New South Wales, between 1982 and 1992. Friesian, Angus, Murray Grey, Guernsey and Hereford breeds were affected. Mean morbidity and case fatality rates in affected herds were 7% (65 of 889) and 69%, respectively. Signs of pruritic dermatitis, illthrift and death were associated with an eosinophilic granulomatous inflammation of many organs, particularly involving the renal cortex, dermis, myocardium, adrenal glands, lymph nodes and hepatic portal triads.  相似文献   

Preliminary study of ovarian activity in fillies treated with a GnRH vaccine   总被引：1，自引：0，他引：1  

U. TSHEWANG  KF DOWSETT  LM KNOTT  TE TRIGG 《Australian veterinary journal》1997,75(9):663-667

Objective To investigate the effects of two doses (200 and 400 mg) of a water-soluble gonadotrophin-releasing hormone vaccine on the ovarian activity of 2-year-old fillies.
Design A controlled vaccination dose rate experiment.
Animals Six 2-year-old Australian Stock Horse fillies were randomly allocated to three treatment groups: unvaccinated controls, those receiving 200 mg of the vaccine and those receiving 400 mg of the vaccine.
Results Ovarian activity of the treated fillies was suppressed at the peak of the breeding season while that of the untreated controls continued normally. The control fillies displayed oestrous activity and behaviour. Suppression of ovarian activity occurred for 25 and 30 weeks in the 200 and 400 mg groups, respectively. These differences were not significant. Ovarian activity ceased 2 to 3 weeks after primary vaccination. Antibody titres were low (330) until after the booster immunisation when they rapidly peaked at 22,000 and 28,000 in the 200 mg and 400 mg groups, respectively. Plasma progesterone concentrations of the treated fillies remained below 3.18 nmol/L while GnRH was suppressed. The vaccine had no significant effect on plasma androstenedione concentrations. Recovery from the effect of the vaccine was associated with development of ovarian follicles, normal oestrous behaviour and ovulation. Three of the four treated fillies and one of the controls conceived during the next breeding season and foaled normally. All the treated fillies conceived and produced normal foals in the following two breeding seasons.
Conclusion Both dose rates suppressed ovarian function and prevented oestrous behaviour. These effects were reversible and the subsequent fertility of the vaccinated fillies was normal.  相似文献   

Granular cell tumour in the bronchus of a horse     

LM GOODCHILD  AJ DART  MB COLLINS  DR HODGSON 《Australian veterinary journal》1997,75(1):16-18

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Using 20-gauge percutaneous peripheral catheters to reliably collect serial 4-mL blood samples from conscious dogs     

KF Elliott  LM Fleeman  JS Rand 《Australian veterinary journal》2010,88(6):215-221

Objective To determine the reliability of collecting serial 4-mL blood samples over 13 h using a 20-gauge, cephalic catheter in conscious dogs. Design Prospective, observational study. Animals Twelve (6 males, 6 females) healthy, neutered, lean dogs. Procedure Percutaneous placement of a 20-gauge, 1.1 × 30 mm intravenous over-the-needle catheter into the cephalic vein was performed in each dog on three occasions. Catheter patency was maintained by intermittent flushing with 0.9% saline and 1 IU/mL heparinised saline solutions. Blood samples (4 mL) were obtained at 10 time-points over 13 h, with close monitoring of the dogs and the indwelling catheters. Blood volume, resistance on aspirating blood sample (minimal or marked resistance) and the methods used to improve blood flow were recorded. Results A high proportion (93%) of blood sample collections achieved the required 4-mL volume, and the remaining samples were greater than 1.5 mL. Of the 358 collections via the cephalic catheter, 311 (87%) were obtained with ‘minimal resistance’. The remaining collections had ‘marked resistance’ (n=47) and were associated with a loose catheter in 11% (5/47) and of unknown cause in 89% (42/47). After ‘marked resistance’ had been encountered and the catheter was flushed with saline and heparin solutions, subsequent sampling with ‘minimal resistance’ was frequently possible from the same catheter. Conclusion Use of a percutaneous, 20-gauge intravenous cephalic catheter allowed reliable serial collection of 4-mL blood samples over 13 h in conscious dogs.  相似文献