The effects of acid rain on nitrogen fixation in Western Washington coniferous forests

We investigated both the current status of N₂ fixation in western Washington forests, and the potential effects of acid rain on this vital process. Even the low concentrations of SO₂ presently found in the Northwest are thought to have an adverse effect on N₂ fixation by limiting the distribution of the epiphytic N₂-fixing lichen, Lobaria pulmonaria, which is found mainly in deciduous forests. A close relative, L. oregana, was found to be the major N₂ fixer in old-growth coniferous forests. It fixes less N₂ following exposure to H₂SO₄ of pH 4 or less. A more serious threat to N₂ fixation than acid rain is the practice of deliberately suppressing red alder to keep it from competing with Douglas fir. Also, L. oregana is a late successional species and does not develop in forests where short cutting cycles are practiced.     

Growth and survival of larval and juvenile cobia Rachycentron canadum in a recirculating raceway system

Cobia Rachycentron canadum is a fast-growing, pelagic marine species that has recently attracted aquaculturists in both the research and commercial sectors. The typical method of grow-out for this species is in outdoor systems where production is limited to locations and seasons conducive for adequate growth and survival. Expanding the culture of cobia to indoor recirculating aquaculture systems (RAS) would allow for the production of fingerlings throughout the year and extend production to cooler regions. Two rearing trials were conducted to examine the growth and survival of cobia from hatching through 4 (trial 1, T1) or 35 (trial 2, T2) g in RAS. Cobia larvae were reared in circular tanks placed in a raceway to control water temperature and quality. During early juvenile grow-out, fish were transferred without grading to a second raceway on 29 dph (T1) or over a period of grading from 29–43 dph (T2). Larval growth (1–22 dph) measured as standard length was similar for both trials ranging from  3.9 to 14.7 mm. However, larval growth measured as wet weight (0.033 g, T1; 0.026 g, T2) or dry weight (5.7 mg, T1; 3.9 mg, T2) was significantly greater on 22 dph during T1 as was the ratio between myotome height and standard length. These differences may have resulted from an increase in initial densities from 8.7 larvae l^− 1 (T1) to 14.7 larvae l^− 1 (T2) which apparently caused an increase in food competition and overall aggression. During juvenile grow-out, cobia reached 4.0 g on 43 dph in T1 and 35.4 g on 71 dph in T2 matching weights achieved during grow-out in outdoor ponds. Over the course of both trials, survival was similar to that reported in outdoor ponds. Mean survival (± S.D.) during the early rearing phase (hatching through 29 or 43 dph) averaged 13.2 ± 3.2 % and 10.4 ± 3.2 % corresponding to final densities of 0.9 ± 0.2 and 1.2 ± 0.4 fish/l for T1 and T2, respectively. During the first grow-out phase (29–43 dph), survival of fish moved into the open raceway was 64.5% in T1 and 88.7 % in T2. Survival of cobia during the second grow-out phase (43–71 dph) for T2 was 92.5%. The results of this study indicate that cobia can be successfully cultured in indoor systems from hatching through at least 35 g without negatively affecting growth or survival.     

Lipid Nutrition and Feeding of Cobia Rachycentron canadum Larvae

This study examined the fatty acid composition of cobia Rachycentron canadum eggs and yolksac larvae, as well as the ovaries of wild caught females as an initial guide to lipid nutritional requirements. A 2-wk feeding study also was conducted to investigate the effectiveness of four dietary treatments on the growth and survival of cobia larvae. Cobia eggs in the tailbud stage contained 31.4 ± 1.3 μg lipid/egg. After hatching, the amount of lipid decreased significantly (P < 0.05) from 28.3 ± 0.3 to 23.2 ± 0.1 μg lipid/larvae during the yolksac larval stage (days 1 to 3 after hatching). Ovaries from wild caught adults and captive spawned eggs and yolksac larvae contained high levels of PUFAs with docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and arachidonic acid (ARA) accounting for approximately 80% of the total suggesting that cobia larvae may have a high dietary requirement for these fatty acids. For the feeding study, larvae were fed: 1) Artemia only; 2) enriched rotifers for 1 d only + microparticulate diet (day 313); 3) enriched rotifers for 3 d (day 3–5) + Artemia (day 3–13); and 4) enriched rotifers for 6 d (day 3–8) + Artemia (day 3–13). Cobia larvae began feeding on rotifers 3 d after hatching and on newly hatched Artemia nauplii by the fifth day following the onset of exogenous feeding (day 7). On day 7, no differences in larval growth were found among larvae fed rotifers for 3 versus 6 d, whereas larvae fed only Artemia or rotifers for I d followed by microparticulate diet were significantly smaller (P < 0.05) and did not survive beyond day 9 and 13, respectively. The results of the feeding study indicate that cobia larvae require rotifers for a minimum of 4 d following the onset of exogenous feeding.     

Development and characterization of mouse monoclonal antibodies reactive with chicken CD80

This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD80 (chCD80). A recombinant plasmid containing a chCD80/horse IgG4 fusion gene was constructed and expressed in CHO cells to produce recombinant chCD80/IgG4 protein. Chicken CD80 was purified from the chCD80/IgG4 fusion protein following enterokinase digestion, and used to immunize BALB/c mice, resulting in 158 hybridomas that produced mAbs against chCD80. Three mAbs with high binding specificity for recombinant chCD80/IgG4-transfected CHO cells were identified by flow cytometry, and one of these (#112) was selected for further characterization. Immunoprecipitation of CD80/IgG4-CHO cell extract, or lipopolysaccharide (LPS)-treated monocytes identified 35.0 kDa proteins. Immunohistochemical analysis revealed chCD80-expressing cells exclusively in the bursal follicles at the outer portion of the cortex, and throughout the red pulp and the outer boundary of the white pulp in the spleen. By immunofluorescence microscopy, chCD80 was observed on intestinal dendritic cells. LPS treatment of bursa or spleen monocytes for 24 or 48 h increased chCD80 expression. Finally, addition of chCD80 mAb to Con A-stimulated spleen cells inhibited the expression of major histocompatibility complex class II antigens and IL-2-driven proliferation of lymphoblast cells. In summary, these chCD80 mAbs will serve as valuable immunological reagents for basic and applied poultry immunology research.     

Development and characterization of mouse monoclonal antibodies reactive with chicken interleukin-2 receptor αlpha chain (CD25)

This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD25 (chCD25), the alpha chain of the interleukin-2 (IL-2) receptor. A recombinant chimeric chCD25/IgG4 fusion protein was expressed in Chinese hamster ovary (CHO) cells and isolated from spent cell culture medium by protein G affinity chromatography. Purified chCD25 protein was used to immunize mice, from which 54 stable hybridomas secreting chCD25 mAbs were produced. Two mAbs, chCD25-32 and chCD25-54, with high binding affinity for chCD25-expressing CHO cells were selected for further characterization. By flow cytometry, both mAbs detected cells in the spleen, bursa of Fabricius, intestinal duodenum, and immunostained established chicken T cell, B cell, and macrophage cell lines. Both mAbs reacted with a 55 kDa protein on Western blots of lysates from concanavalin A (Con A)-stimulated spleen mononuclear cells. Intraperitoneal injection of chickens with bacterial lipopolysaccharide increased the percentage of chCD25(+) spleen cells by approximately 4-fold compared with untreated animals. In vitro stimulation of spleen cells with Con A increased the percentage of chCD25(+) cells by up to 50-fold compared with cells treated with medium alone. Finally, the chCD25-32 mAb suppressed IL-2-driven spleen cell proliferation and reduced IL-2-induced nitric oxide production. These mAbs may be useful for future investigation of chicken regulatory T cells.     

Chronic hepatitis in Labrador Retrievers: clinical presentation and prognostic factors

BACKGROUND: An increased incidence of chronic hepatitis has been reported in Labrador Retrievers. HYPOTHESIS: A breed associated hepatopathy occurs in Labrador Retrievers. ANIMALS: Twenty-four client-owned Labrador Retrievers. METHODS: Medical records of dogs with histopathologic confirmation of chronic hepatitis were retrospectively reviewed. A clinical score based on clinical signs and the results of biochemical tests was generated for each dog. Hepatic biopsy specimens were scored for disease activity, fibrosis, and copper accumulation. RESULTS: The median age was 9.3 years (range, 3.9-14.0 years). Clinical signs included inappetence, vomiting, lethargy, and weight loss. All dogs had increases in serum activity of one or more hepatobiliary enzyme. Hyperbilirubinemia and hypoalbuminemia were present in 45% and 21% of dogs, respectively. The median clinical score was 2.9, with a range of 0-8. The median histopathology activity and the fibrosis scores were 3.5 (range, 1-6) and 3.0 (range, 0-4), respectively. Rhodanine-positive copper staining was present in 15 of 17 biopsy specimens, with a median score of 2.0 (range, 0-3). Median survival was 374 days (range, 1-2645 days). A prolonged prothrombin time (P = .013) and thrombocytopenia (P = .041) were associated with survival < 2 months. The presence of anorexia (P = .049), hypoglobulinemia (P = .045), or prolonged partial thromboplastin time (P = .033) were associated with shorter overall survival times. The clinical score correlated with survival time (P = .030) and histopathologic staging (P = .049). CONCLUSIONS AND CLINICAL IMPORTANCE: A progressive hepatopathy in Labrador Retrievers in this study was marked by chronic inflammation, fibrosis, and copper accumulation. A clinical scoring system that correlates with survival time may be useful as a noninvasive method to predict prognosis.     

Liver function in cats with hyperthyroidism before and after 131I therapy

BACKGROUND: The clinical significance of high serum concentration or activity of markers of liver damage in cats with hyperthyroidism is unknown. OBJECTIVE: To evaluate serum markers of liver function and damage, and ultrasonographic changes in cats with hyperthyroidism and with high liver enzymes, and to determine if abnormalities resolve after treatment with 131I. ANIMALS: Nineteen cats with hyperthyroidism (15 with high serum activities of liver enzymes) and 4 age-matched healthy control cats. METHODS: Serum bile acids, albumin, ammonia, cholesterol, and blood urea nitrogen concentrations, and activities of liver-derived enzymes, and blood glucose concentrations were measured before and after 131I therapy. These values were compared with those of cats that were euthyroid. In addition, gross liver parenchymal changes detected by abdominal ultrasonographic examination, before and after 131I therapy were evaluated. RESULTS: High serum liver enzyme activities were not associated with abnormalities in hepatic parenchyma and liver functional variables, regardless of the degree of increase. Serum liver enzyme activities return to normal after control of hyperthyroidism with 131I therapy. Cats with hyperthyroidism have a significantly higher serum fasting ammonia concentration than cats who were euthyroid (P = .019). Cats with hyperthyroidism also have significantly lower serum cholesterol (P = .005) and glucose (P = .002) concentrations before compared with after 131I therapy. Nine of 19 cats with hyperthyroidism had trace ketonuria. CONCLUSIONS AND CLINICAL IMPORTANCE: These results demonstrate that extensive examination for hepatobiliary disease in most cats with hyperthyroidism is unnecessary.     

Effect of fentanyl on visceral and somatic nociception in conscious horses

BACKGROUND: Transdermal fentanyl is used clinically in horses based on pharmacokinetic data and antinociceptive effects documented in other species. HYPOTHESIS: Fentanyl IV administration increases both visceral and somatic nociceptive threshold in conscious horses. ANIMALS: Six clinically normal horses, each fitted with a permanent gastric cannula. METHODS: Visceral nociception was evaluated with 2 methods of threshold detection--olorectal distention and duodenal distention. Somatic nociception was assessed by measurement of thermal threshold. Fentanyl was administered as an increasing stepwise infusion followed by a continuous-rate infusion for a total of 2 hours. There were 4 doses of fentanyl and 1 dose each of saline and xylazine administered to each horse. Serum fentanyl concentrations were measured and the resulting data were used to determine pharmacokinetic parameters for each horse. All data were analyzed by means of a 3-factor analysis of variance followed by either a simple t test or a Bonferroni t test for multiple comparisons. RESULTS: Fentanyl administration did not result in significant changes in duodenal or colorectal distention threshold. Thermal threshold showed an increased trend at the 15-minute time point for the highest fentanyl group only, with a corresponding mean serum fentanyl concentration of 7.82 +/- 2.10 ng/mL. Two horses in this group became agitated and tachycardic during the first 15 minutes of the infusion. CONCLUSIONS AND CLINICAL IMPORTANCE: Fentanyl did not produce a significant antinociceptive effect at the doses used, 2 of which resulted in serum concentrations above the nociceptive threshold in other species.