Establishment of a time-resolved fluoroimmunoassay for measuring plasma insulin-like growth factor I (IGF-I) in fish: effect of fasting on plasma concentrations and tissue mRNA expression of IGF-I and growth hormone (GH) in channel catfish (Ictalurus punctatus)

A time-resolved fluoroimmunoassay (TR-FIA) was established and validated that allows for the determination of plasma concentrations of insulin-like growth factor I (IGF-I) in three domestically cultured fishes: channel catfish (Ictalurus punctatus), hybrid striped bass (Morone chrysopsxM. saxatilis), and rainbow trout (Oncorhynchus mykiss). Sensitivity of the assay was 0.20 ng/ml. Intra- and inter-assay coefficients of variation (CV) were <7 and <12%, respectively. Serial dilutions of plasma from each species were parallel to the standard curve. Recovery of IGF-I from spiked plasma samples was >90% for all three species of fishes. The IGF-I TR-FIA was biologically validated via its use to determine the effect of fasting on circulating IGF-I levels in channel catfish. Fasting-induced changes in plasma growth hormone (GH), hepatic IGF-I mRNA expression, and pituitary GH mRNA expression were also determined. Fasted channel catfish lost 5.6 and 15.6% body mass after 2 and 4 weeks of fasting, respectively. Plasma IGF-I concentrations were depressed (P<0.05) relative to fed controls following 2 and 4 weeks of fasting. Plasma GH concentrations were not different (P>0.05) in fasted fish after 2 weeks, but significantly increased (P<0.05) by 4 weeks of fasting. Hepatic IGF-I mRNA expression after 2 and 4 weeks of fasting was reduced (P<0.05) relative to fed controls. Pituitary GH mRNA expression was similar (P>0.05) between 2-week-fasted catfish and fed controls, but was increased (P<0.05) in 4-week-fasted catfish. The IGF-I TR-FIA was sensitive, accurate, and precise for all three species of fishes, and provided a low-cost, and non-radioisotopic method for quantifying plasma IGF-I levels in fed and fasted channel catfish.     

Comparison of gastritis and gastric epithelial proliferation in Helicobacter heilmannii-infected nude and BALB/c mice

Gastric mucosal hypertrophy/nodular hyperplasia occurs as a consequence of Helicobacter infection in mice and humans. The pathogenesis of this hyperplastic response is not understood. To determine the role of host cellular immunity in gastric mucosal hypertrophy/hyperplasia, 6-8-week-old female euthymic BALB/c (n = 30) or NIH athymic nude (n = 40) mice were inoculated with H. heilmannii. Equal numbers of uninoculated mice of each strain served as controls. Mice from each group were euthanatized at 0.5, 6, 12, and 18 months postinoculation (PI). Lymphoplasmacytic gastritis and lymphoid follicle development were less severe in nude mice than in euthymic mice at <6 months PI. The prevalence of gastritis at 0.5, 6, 12, and 18 months PI was 0%, 17%, 67%, and 88%, respectively, in infected nude mice and 33%, 83%, 71%, and 100%, respectively, in infected BALB/c mice. CD4+ T cells in infected nude mice were evident at > or =6 months PI but were less numerous than in infected BALB/c mice at comparable time intervals. However, numbers of CD4+ T cells increased substantially throughout the experiment in infected BALB/c mice. The prevalence of nodular mucosal hyperplasia at 0.5, 6, 12, and 18 months PI was 0%, 0%, 33%, and 20%, respectively, in infected nude mice and 0%, 33%, 80%, and 80%, respectively, in infected BALB/c mice. Nodular hyperplasia occurred in association with the appearance of chronic lymphoplasmacytic inflammation and CD4+ T cells at 12 and 18 months PI in nude mice. H. heilmannii-associated gastritis and mucosal remodeling is reduced in immunodeficient mouse strains lacking normal CD4+ T cell numbers as compared with the response in immunocompetent mice. Additionally, CD4 immunocompetence is an integral aspect of the mucosal hypertrophy/nodular hyperplasia in experimental H. heilmannii-associated disease of mice.     

Serum thyroid hormone concentrations in clinically normal dogs after administration of freshly reconstituted vs previously frozen and stored thyrotropin

Concentrations of serum thyroxine (T4) and 3,3',5-triiodothyronine (T3) were determined in 7 clinically healthy adult dogs before and after administration of freshly reconstituted thyrotropin (TSH) and TSH that had been previously reconstituted and frozen for 1, 2, and 3 months. The 4 TSH response tests were performed at 30-day intervals by collecting blood samples for serum T4 and T3 determinations before and 4 and 6 hours after IV administration of TSH (0.1 U/kg of body weight). Baseline serum concentrations of T4 and T3 were similar at each of the 4 sample collection times over the 3-month period of the study. Mean serum concentrations of T4 and T3 increased significantly (P less than 0.01) over baseline values after administration of freshly reconstituted TSH or TSH that had been previously frozen for 1, 2, or 3 months. Significant difference was not found in the mean post-TSH serum T4 or T3 concentration after injection of freshly reconstituted TSH or TSH that had been previously frozen for 1, 2, or 3 months. In 2 of the 7 dogs, mild reactions--mild ataxia and weakness--were observed during the last of the series of TSH response tests (ie, after IV administration of TSH that had been previously frozen for 3 months). Results of this study suggest that for use in dogs, reconstituted TSH stored at -20 C maintains adequate biological activity for at least 3 months. The ability to store reconstituted TSH for a longer period than the recommended 48 hours represents an economic advantage, because it allows clinicians to perform more TSH response tests per vial of TSH.     

Primary Hypoadrenocorticism in Ten Cats

Primary hypoadrenocorticism was diagnosed in ten young to middle-aged cats of mixed breeding. Five of the cats were male, and five were female. Historic signs included lethargy (n = 10), anorexia (n = 10), weight loss (n = 9), vomiting (n = 4), and polyuria (n = 3). Dehydration (n = 9), hypothermia (n = 8), prolonged capillary refill time (n = 5), weak pulse (n = 5), collapse (n = 3), and sinus bradycardia (n = 2) were found on physical examination. Results of initial laboratory tests revealed anemia (n = 3), absolute lymphocytosis (n = 2), absolute eosinophilia (n = 1), and azotemia and hyperphosphatemia (n = 10). Serum electrolyte changes included hyponatremia (n = 10), hyperkalemia (n = 9), hypochloremia (n = 9), and hypercalcemia (n = 1). The diagnosis of primary adrenocortical insufficiency was established on the basis of results of adrenocorticotropic hormone (ACTH) stimulation tests (n = 10) and endogenous plasma ACTH determinations (n = 7). Initial therapy for hypoadrenocorticism included intravenous administration of 0.9% saline and dexamethasone and intramuscular administration of desoxycorticosterone acetate in oil. Three cats were euthanatized shortly after diagnosis because of poor clinical response. Results of necropsy examination were unremarkable except for complete destruction of both adrenal cortices. Seven cats were treated chronically with oral prednisone or intramuscular methylprednisolone acetate for glucocorticoid supplementation and with oral fludrocortisone acetate or intramuscular injections of repository desoxycorticosterone pivalate for mineralocorticoid replacement. One cat died after 47 days of therapy from unknown causes; the other six cats are still alive and well after 3 to 70 months of treatment.     

Observations on the abundance of the exotic cichlid Sarotherodon mossambicus (Peters) in relation to fluctuations in the water-level in a man-made lake in Sri Lanka

Abstract. The fishery of Parakrama Samudra, an ancient man-made lake of 2662 ha in Sri Lanka (8°-10°N; 80°-82°E), as in most other man-made reservoirs in the island, is dominated by the exotic cichlid Sarotherodon mossambicus (Peters), introduced in 1952. The fluctuations in catch per unit/fisherman over the period 1960 to 1982 are shown to be dependent on the yearly fluctuations in the water-level, the changes in the water-level effects being manifested in the fishery after a period of 3 years. The above results were arrived at by auto-correlation analysis of yearly catch per unit effort against the fluctuation of the water level at the main sluice. The changes in yield in a particular year are correlated to the changes in the water level and are described by the equation;
where Y = yield in kg fisherman^-1 month^-1 in year t and X = fluctuations in the water-level in year t - 3. The available growth rates in natural and semi-natural impoundments also indicate that the size at which S. mossambicus enter the fishery in Parakrama Samudra corresponds to their size in the third year of life (age 2+). The significance of the above observations is discussed on the basis of available knowledge on the reproductive cycle and the nesting requirements of S. mossambicus which are achieved when significant fluctuations in the reservoir level take place by making available optimal water depths in areas with a suitable substratum. The possible use of the above criteria as a management measure is also briefly discussed in the light of possible irrigational and fisheries conflicts in the utilization of the reservoir resources.     

Phenology and spatial and temporal distribution of larval fishes in a partially channelized warmwater stream

We studied the phenology and spatial and temporal dynamics of larval fish abundance with light traps from 5 locations along the length of Luxapalilia Creek, a partially channelized warmwater stream in northeastern Mississippi over 2 spawning seasops (March through August 1992 and 1993). The dominant, early spring spawners were catostomids (early-April to late-May) and darters (mid-March to early-June) whereas summer spawners were mainly cyprinids (mid-May to August) and centrarchids (late-May to August). The relative abundance of larvae collections differed based on: 1) site, 2) location, 3) date, 4) year, and 5) in larval stage. Total numbers of larvae and CPUE were high in mid- to late summer in downstream locations in both years when: 1) temperatures were high, 2) water velocities were low, and 3) depths were shallow compared to upstream locations. More mature larvae (meso- and metalarvae) were more common in the downstream locations whereas less mature larvae (yolk-sac and protolarvae) were more abundant at the upstream locations. The downstream channelized portion of Luxapalila Creek resembled a lentic habitat with littoral zones characterized by slow water velocities, shallow depths, fine sediments, and vegetation. Larvae of darters (Etheostorna and Pcrcina), which require gravel bars and swift currents, were not collected in great numbers downstream, although members of the Ictiobinae (Carpoides and Ictiobus), cyprinidae and centrarchidae (Lepomis spp., Pomoxis spp. and Micropterus spp.) were more abundant downstream than upstream. This suggests that the natural, upstream locations acted as sources of larvae for the channelized downstream location.