Development and validation of a range of endogenous controls to support the implementation of practical Taqman real-time PCR-based surveillance for fish diseases within aquaculture

The use of Taqman real-time PCR-based technology has recently become more frequent in the detection of pathogens in the aquaculture industry. This interest has necessitated the development of robust and reliable pathogen-detection assays. The development of a range of endogenous control assays to be run alongside these diagnostic assays works to further increase confidence in the latter. This study describes the design of a range of endogenous control assays based on the elongation factor 1-α (EF1-α) gene specific to a range of fish species including Atlantic salmon, Salmo salar; rainbow trout, Oncorhynchus mykiss; brown trout, Salmo trutta; cod, Gadus morhua; haddock, Melanogrammus aeglefinus; saithe, Pollachius virens; whiting, Merlangius merlangus; Norway pout, Trisopterus esmarkii; carp (family Cyprinidae), roach, Rutilus rutilus; European eel, Anguilla anguilla; and herring, Clupea harengus, as well as a number of fish cell lines. Evidence is provided of the validation of these assays for specific species, a range of tissue types and cell lines as well as an example of the potential uses of these assays.     

Effect of Initial GnRH and Duration of Progesterone Insert Treatment on the Fertility of Lactating Dairy Cows

The study compared response to prostaglandin F2α (PG), synchrony of ovulation and pregnancy per AI (P/AI) in a 5‐ vs a 7‐day Ovsynch + PRID protocol and investigated whether the initial GnRH affects P/AI in lactating dairy cows. Two hundred and seventy‐six cows (500 inseminations) were assigned to one of four timed‐AI (TAI) protocols: (i) PRID‐7G; 100 μg GnRH im, and a progesterone‐releasing intravaginal device (PRID) for 7 days. At PRID removal, PG (500 μg of cloprostenol) was given im. Cows received the second GnRH treatment at 60 h after PRID removal and TAI 12 h later. (ii) PRID‐5G; as PRID‐7G except the duration of PRID, treatment was 5 days and PG was given twice (12 h apart). (iii) PRID‐7NoG; as PRID‐7G except the initial GnRH, treatment was omitted. (iv) PRID‐5NoG; as PRID‐7NoG except the duration of PRID, treatment was 5 days. Response to treatments and pregnancy status at 32 and 60 days after TAI was determined by ultrasonography. The percentage of cows ovulating before TAI was greatest in PRID‐7G (17.1%), and the percentage of cows that did not have luteal regression was greatest in PRID‐5G (9.5%). The overall P/AI at 32 and 60 days did not differ among TAI protocols. However, during resynchronization, cows subjected to the 5‐day protocols had greater (p < 0.05) P/AI (45.3% vs 33.6%) than cows subjected to the 7‐day protocols. Pregnancy loss between 32 and 60 days tended (p = 0.10) to be greater in cows that did not receive initial GnRH (14.8%) compared to those that received GnRH (8.2%). In conclusion, the PRID‐5G protocol resulted in fewer cows responding to PG, but P/AI did not differ among TAI protocols. A 5‐day protocol resulted in more P/AI in resynchronized cows, and cows that did not receive initial GnRH tended to experience more pregnancy losses.     

Acute phase response in calves following infection with Pasteurella haemolytica, Ostertagia ostertagi and endotoxin administration

The concentrations of bovine acute phase proteins were monitored in plasma following experimental infection with Pasteurella haemolytica and Ostertagia ostertagi and after endotoxin administration. Raised levels of haptoglobin, alpha 1 proteinase inhibitor and seromucoid were detected after pasteurella infection and endotoxin administration. Ceruloplasmin levels increased after endotoxin administration but not during pasteurella infection. Raised levels of the four acute phase proteins were found in eight of 19 calves infected with ostertagia but showed a variable pattern and did not correlate with plasma pepsinogen increases. Bovine alpha 1 antichymotrypsin and alpha 2 macroglobulin were identified as acute phase reactants.     

Bragg diffraction from crystallized ion plasmas

Single crystals of a one-component plasma were observed by optical Bragg diffraction. The plasmas contained 10(5) to 10(6) single-positive beryllium-9 ions (9Be+) at particle densities of 10(8) to 10(9) per cubic centimeter. In approximately spherical plasmas, single body-centered cubic (bcc) crystals or, in some cases, two or more bcc crystals having fixed orientations with respect to each other were observed. In some oblate plasmas, a mixture of bcc and face-centered cubic ordering was seen. Knowledge of the properties of one-component plasma crystals is required for models of white dwarfs and neutron stars, which are believed to contain matter in that form.     

Hydrogen radicals, nitrogen radicals, and the production of O3 in the upper troposphere

The concentrations of the hydrogen radicals OH and HO2 in the middle and upper troposphere were measured simultaneously with those of NO, O3, CO, H2O, CH4, non-methane hydrocarbons, and with the ultraviolet and visible radiation field. The data allow a direct examination of the processes that produce O3 in this region of the atmosphere. Comparison of the measured concentrations of OH and HO2 with calculations based on their production from water vapor, ozone, and methane demonstrate that these sources are insufficient to explain the observed radical concentrations in the upper troposphere. The photolysis of carbonyl and peroxide compounds transported to this region from the lower troposphere may provide the source of HOx required to sustain the measured abundances of these radical species. The mechanism by which NO affects the production of O3 is also illustrated by the measurements. In the upper tropospheric air masses sampled, the production rate for ozone (determined from the measured concentrations of HO2 and NO) is calculated to be about 1 part per billion by volume each day. This production rate is faster than previously thought and implies that anthropogenic activities that add NO to the upper troposphere, such as biomass burning and aviation, will lead to production of more O3 than expected.     

Sensitivity of boreal forest carbon balance to soil thaw

We used eddy covariance; gas-exchange chambers; radiocarbon analysis; wood, moss, and soil inventories; and laboratory incubations to measure the carbon balance of a 120-year-old black spruce forest in Manitoba, Canada. The site lost 0.3 +/- 0.5 metric ton of carbon per hectare per year (ton C ha-1 year-1) from 1994 to 1997, with a gain of 0.6 +/- 0.2 ton C ha-1 year-1 in moss and wood offset by a loss of 0.8 +/- 0.5 ton C ha-1 year-1 from the soil. The soil remained frozen most of the year, and the decomposition of organic matter in the soil increased 10-fold upon thawing. The stability of the soil carbon pool ( approximately 150 tons C ha-1) appears sensitive to the depth and duration of thaw, and climatic changes that promote thaw are likely to cause a net efflux of carbon dioxide from the site.