Variability of serum bile acid concentrations over time in dairy cattle, and effect of feed deprivation on the variability.

Twelve nonlactating dairy cows, free of signs of liver disease and with normal serum activities of liver-derived enzymes and normal liver biopsy tissue, were examined over a 72-hour period for serum total bile acid concentrations. The cattle were fed hay twice daily, and blood samples were obtained every hour for 24 hours, every other hour for 24 hours, then every hour for 24 hours. After 3 weeks, the study was repeated on 6 of the cattle, thus providing data for eighteen 72-hour periods. Serum bile acid concentration varied greatly over the 72 hours, with the range being from one third to 3 times the median. There were variations by as much as 60 mumol/L from 1 hour to the next. After another 3 weeks, 8 of the cattle were deprived of hay for 48 hours and then fed hay morning and afternoon of the third (last) day of the study. There was no significant reduction in bile acid concentration after withholding the hay, but the variability was reduced (P = 0.02) during the last 20 hours of the hay-deprivation period. In 3 ancillary studies, serum bile acid concentrations were examined over a 48-hour period in 2 cows in early lactation, 3 cows in midlactation, and two 6-month-old heifers. The cows were fed hay and grain twice daily, and the heifers were fed only hay twice daily. In comparison with values for the 12 nonlactating cows fed hay twice daily, mean serum bile acid concentration in the recently freshened cows was significantly (P < 0.002) higher (62.9 vs 22.0 mumol/L).(ABSTRACT TRUNCATED AT 250 WORDS)     

Multiresidue assay for benzimidazole anthelmintics by liquid chromatography and confirmation by gas chromatography/selected-ion monitoring electron impact mass spectrometry

A multiresidue method utilizing all-disposable labware has been developed for 8 benzimidazole anthelmintics from ovine, bovine, and swine muscle and liver tissues. After an initial extraction with ethyl acetate and subsequent evaporation, a 3-component extraction using hexane, ethanol, and 0.2N HCl was used for final cleanup. Clean extracts were produced for separation and determination by reverse-phase liquid chromatography at 298 nm, using methanol and aqueous buffer as mobile phase. A synthesized internal standard, 2-(n-butylmercapto)benzimidazole, was used for quantitation of all drugs. Results are included along with statistical information verifying the performance of the method. Spiked control tissues and incurred drug tissues were used for an intralaboratory study with a concentration range of 50-1470 ppb. A series of standard curves at 0, 50, 100, and 200 ppb were analyzed. Overall recovery at the 100 ppb level averaged 92% (CV 8%) in liver tissues, across all 3 species and 88% (CV 5%) in muscle tissues across all 3 species. Results were confirmed by gas chromatography/mass spectrometry with acid hydrolysis of the remaining extract in 2N HCl followed by re-extraction of the amine and derivatization to the tert-butyldimethylsilyl derivative. The anthelmintics were identified by gas chromatography/selected ion monitoring electron-impact mass spectrometry. Ion ratio measurements were taken and compared to standard material. CVs averaged 10% or less for all drugs tested.     

A framework for evaluation of on-farm mastitis diagnostics in Australia

Numerous culture-based diagnostics are available on the Australian and international markets for on-farm detection of bacterial pathogens in milk. Use of such diagnostics may provide an opportunity to improve the prudent use of antimicrobials in udder health management. Farms are low-resource settings in terms of diagnostic microbiology capacity. The World Health Organisation has identified criteria for the evaluation of diagnostic tests in low resource settings based on Accuracy, Sensitivity, Specificity, User-friendliness, being Rapid or Robust, Equipment-free and being Deliverable (ASSURED). Here, we review how those criteria can be interpreted in the context of microbiological diagnosis of mastitis pathogens, and how on-farm diagnostics that are currently available in Australia perform relative to ASSURED criteria. This evaluation identifies multiple trade-offs, both with regard to scientific criteria and with regards to convenience criteria. More importantly, the purpose of testing may differ between farms, and test performance should be evaluated relative to its intended use. The ability of on-farm mastitis diagnostics to inform mastitis treatment decision-making in a timely and cost-effective manner depends not just on test characteristics but also on farm-specific pathogen prevalence, and on the farm enterprise's priorities and the farm manager's potential courses of action. With most assay evaluations to date conducted in professional laboratories, there is a surprising dearth of information on how well any of the diagnostic tests perform on-farm and, indeed, of the on-farm decision-making processes that they aim to inform.     

Extraction, processing, and storage effects on curcuminoids and oleoresin yields from Curcuma longa L. grown in Jamaica

Aromatic diarylheptanoid compounds from Curcuma longa Linn grown in Jamaica were quantified by UV-vis spectrophotometry and high-performance liquid chromatographic (HPLC) analyses. The oleoresin yields from ethanolic extracts were quantified and evaluated with regard to the effects of the type of postharvesting process and the type of extraction method conducted on the plant material. Fresh samples that were hot solvent extracted provided the highest oleoresin yields of 15.7% +/- 0.4 ( n = 3), and the lowest oleoresin yields of 7.8% +/- 0.2 ( n = 3) were from the dried milled samples that were cold solvent extracted. Data from the ASTA spectrophotometer assay confirmed that dried samples contained the highest curcuminoid content of 55.5% +/- 2.2 ( n = 6) at the fifth month of storage, and the fresh samples showed a curcuminoid content of 47.1% +/- 6.4 ( n = 6) at the third month of storage. A modified HPLC analysis was used to quantify curcumin content. Data from the HPLC analysis confirmed that the dried treated, hot extracted, room temperature stored samples had the highest curcumin content of 24.3%. A novel high-performance thin layer chromatography (HPTLC) method provided a chemical fingerprint of the C. longa with the use of a commercial curcumin standard.     

Foliar nematode,Litylenchus crenatae ssp. mccannii,population dynamics in leaves and buds of beech leaf disease‐affected trees in Canada and the US

A foliar nematode, Litylenchus crenatae ssp. mccannii, is associated with beech leaf disease (BLD) symptoms. Information about the types of tissues parasitized and how nematode populations fluctuate in these tissues over time is needed to improve surveys as well as understand the nematodes role in BLD. During this study, the nematode was detected throughout the known range of BLD by researchers at both Canadian and US institutions using a modified pan method to extract nematodes. Monthly collections of symptomatic and asymptomatic leaves during the growing season (May–October), and leaves and buds between growing seasons (November–March), revealed that nematodes were present in all tissue types. Progressively larger numbers of nematodes were detected in symptomatic leaves from Ohio and Ontario, with the greatest detections at the end of the growing season. Smaller numbers of nematodes were detected in asymptomatic leaves from BLD‐infected trees, typically at the end of the growing season. The nematode was detected overwintering in buds and detached leaves. The discovery of small numbers of nematodes in detached leaves at one location before BLD was detected indicates that nematodes may have been present before disease symptoms were expressed. Other nematodes, Plectus and Aphelenchoides spp., were infrequently detected in small numbers. Our findings support the involvement of the nematode in BLD and indicate that symptoms develop only when certain requirements, such as infection of buds, are met. We also found that the nematode can be reliably detected in buds and leaves using the modified pan extraction method.     

Isolation and characterisation of cervine herpesvirus-1 from red deer semen

AIM: This communication describes the isolation of herpesvirus during routine export examination of semen collected from red deer stags in New Zealand. METHODS: Virus isolation was carried out using bovine embryonic lung (BEL) cells and viruses were characterised by direct immunofluorescense, restriction-fragment-length polymorphism analysis (RFLP), polymerase chain reaction (PCR) analysis and nucleotide sequencing. RESULTS: Herpesvirus was isolated from red deer semen on 2 different occasions from different animals. In both cases the virus was identified as cervine herpesvirus-1 (CvHV-1), based on RFLP, PCR and sequence analysis. Nucleotide sequence analysis of the glycoprotein-D gene showed 99.7% homology to the Banffshire strain of CvHV-1 and 89.5%, 89.2%, 85.3% and 79.6% homology to bovine herpesvirus 1.2 (BoHV-1.2), bovine herpesvirus 1.1 (BoHV-1.1), cervine herpesvirus-2 (CvHV-2) and caprine herpesvirus-1 (CpHV-1), respectively. CONCLUSION: This is the first time that CvHV-1 has been isolated in New Zealand. Its inclusion in serological surveys will allow the prevalence of CvHV-1 in the red deer population to be assessed in this country. The clinical significance of CvHV1 infection in New Zealand red deer herds has yet to be determined.     

Untersuchungen zum endophytischen Befall von Fusarium proliferatum (Matsushima) Nirenberg in geernteten Stangen von Spargel (Asparagus officinalis L.)

Zusammenfassung Erstmalig wurden mit den vorliegenden Untersuchungen Spargelstangen zur Haupterntezeit auf endophytischen Pilzbefall untersucht. Sie zeigen, dass im Ernteprodukt zwar Fusarium proliferatum als potenzieller Mykotoxinbildner zu finden ist. Eine mögliche natürliche Kontamination mit Fumonisinen bestätigte sich nicht. Von den mit F. proliferatum infizierten Stangen wies nur eine Stange mit grau-rosa-orange farbenen Gewebeveränderungen an der Basis sichtbare Symptome auf. Allgemeine Rückschlüsse auf eine mögliche Gefährdung oder Nichtgefährdung des Verbrauchers beim Verzehr von mit F. proliferatum kontaminierten, symptomlosen Stangen können aus der Analyse nicht gezogen werden. Hierzu müssen weitergehende Untersuchungen zur Wirt-Pathogen-Interaktion erfolgen und die phänotypischen und genotypischen Einflussfaktoren in diesem Prozess noch näher untersucht werden.