Detecting and quantifying the adventitious presence of transgenic seeds in safflower, Carthamus tinctorius L

Safflower ( Carthamus tinctorius L.) is currently being developed as a platform for the production of novel proteins. Methods for detecting and quantifying transgenic safflower are needed to ensure seed quality and to monitor for its adventitious presence. We developed and compared three methods of assaying for transgenic safflower presence in conventional seedlots: field bioassays, enzyme-linked immunosorbent assays (ELISA), and quantitative polymerase chain reaction (Q-PCR). Limits for reliable quantification for both ELISA and Q-PCR are approximately 0.1%, although levels at least as low as 0.02% can be detected by Q-PCR. Levels of quantification for the field bioassay are limited only by space and resources available. Multiple sampling methods to detect and quantify transgenic safflower presence at levels lower than 0.1% were used on field collected samples from a pollen outcrossing experiment to quantify the adventitious presence of transgenic safflower. Taking into account the potential utility and relative advantages or disadvantages of each detection method, it is recommended that the initial testing for the adventitious presence of transgenic seed be carried out using an antibody-based test if available and that Q-PCR-based assays to quantify transgenic proportion be used when it is necessary to identify specific transgenic constructs or if antibody-based assays are not readily available.     

Energy dynamics of subyearling Chinook salmon reveal the importance of piscivory to short‐term growth during early marine residence

Variation in prey quantity and quality can influence growth and survival of marine predators, including anadromous fish that migrate from freshwater systems. The objective of this study was to examine the energy dynamics of subyearling Chinook salmon (Oncorhynchus tshawytscha) following freshwater emigration. To address this objective, a population of Chinook salmon and their marine prey were repeatedly sampled from June to September over 2 years in coastal waters off Oregon and Washington. Subyearlings from the same population were also reared under laboratory conditions. Using a bioenergetics model evaluated in the laboratory, we found that growth rate variability in the field was associated more with differences in northern anchovy (Engraulis mordax) consumption and less with variation in diet energy density or ocean temperature. Highest growth rates (2.43–3.22% body weight/day) occurred in months when anchovy biomass peaked, and the timing of peak anchovy biomass varied by year. Our results support a general pattern among subyearling Chinook salmon occurring from Alaska to California that feeding rates contribute most to growth rate variability during early marine residence, although dominant prey types can differ seasonally, annually, or by ecosystem. In the northern California Current, faster growth appears to be associated with the availability of age‐0 anchovy. Identifying factors that influence the seasonal development of the prey field and regulate prey quantity and quality will improve understanding of salmon growth and survival during early marine residence.     

Bronchoalveolar lavage cytology from captive badgers

Background: Bronchoalveolar lavage (BAL) fluid is evaluated for the diagnosis and study of lung disease and airway inflammation. Cytologic profiles for BAL fluid have not been reported for badgers and may be useful in understanding the pathogenesis of pulmonary diseases such as Mycobacterium bovis. Objective: The aim of this study was to evaluate cytologic and microbial findings in BAL fluid from captive European badgers (Meles meles) and identify correlates with the results of concurrently collected blood and fecal samples. Methods: BAL fluid (by a nonbronchoscopic method) and jugular venous blood samples (for routine CBC) were obtained from 23 captive tuberculosis‐free anesthetized badgers on 2 occasions 4 weeks apart. Fecal samples were collected for routine parasitology. Morphologic evaluation and 100‐cell differentials were done on cytocentrifuged BAL specimens. Pellets from centrifuged BAL were aerobically cultured for bacteria. Results: With the 2 BAL samples from each of the 23 badgers combined, the median (range) cell percentages were 73.0% (5–95%) neutrophils, 7.5% (2–16%) macrophages, 8.0% (0–27%) lymphocytes, and 9.5% (0–92%) eosinophils. Macrophages frequently contained silica‐like crystals. Other findings included ciliated epithelial cells, goblet cells, mucus, and Aelurostrongylus sp. larvae. A light growth of Streptococcus, Pasteurella, or Escherichia coli was cultured in 6 badgers. Trypanosoma pestanai were identified in blood from 10 badgers and fecal parasites (mainly coccidia) were found in 20 badgers. No correlation was found between BAL and CBC results and the presence of parasites. Conclusions: The predominance of neutrophils in BAL fluid from badgers differs from the predominance of macrophages found in BAL from other species. This difference may reflect the burrowing lifestyle or the unique immune response of badgers.     

MAGNETIC RESONANCE IMAGING FINDINGS OF DESMOPATHY OF THE COLLATERAL LIGAMENTS OF THE EQUINE DISTAL INTERPHALANGEAL JOINT

We report the use of a low-field magnetic resonance (MR) imaging system for the detection of desmopathy of the collateral ligament of the distal interphalangeal joint and the long-term outcome. Twenty horses were studied and their medical records and MR images were reviewed retrospectively. Long-term follow-up information was obtained by telephonic questionnaires of owners, trainers, or referring veterinarians. Desmopathy of the medial collateral ligament (80%) and enthesopathy of the affected collateral ligament (80%) were common MR imaging features. Treatment consisted of stall rest followed by a rehabilitation period. Additional treatments included shoeing, extracorporeal shock wave therapy, application of a half limb or foot cast, and medication of the distal interphalangeal joint. Twelve (60%) horses returned to their previous level of exercise and maintained their previous level, whereas eight horses had a poor outcome. Low-field MR imaging in the standing patient can be used to detect collateral ligament desmopathy of the distal interphalangeal joint without a need for general anesthesia.     

Beta-amylase degradation by serine endoproteinases from green barley malt

Proteolytic degradation of barley proteins is examined in green (unkilned) malt and germinating seeds from Hordeum vulgare L. cv. Harrington. Zymographic analysis of the Harrington green malt extracts using commercial preparations of barley beta-amylase incorporated as a proteolytic substrate in 2-D SDS gels shows multiple proteolytic activities. A developmental study shows that the several green malt beta-amylase-degrading activities appear at around day 2 of germination. The several activities appear to increase and decrease through 7 days of germination in a coordinated fashion. Gels treated with class-specific proteinase inhibitors show that serine-class proteinase activities are responsible for barley beta-amylase degradation seen on the zymograms. Western blot analysis also shows that proteolytic enzymes recovered from 1-D electrophoretic gels degrade barley beta-amylase, and that the degradation is inhibited by PMSF. This is the first demonstration that malt proteinases are capable of degrading important metabolic enzymes in germinating barley, and the first postulated physiological role for the serine class proteinases in barley malt.