ALBERT

Notes: . Oxytricha strains used in biochemical studies have traditionally been grown in unaerated, unagitated culture tubes or Fernbach flasks. These cultures are limited in volume to about one liter and have a very nonuniform distribution of cells, with the majority of the cells at the very top or bottom of the medium. We have found conditions in which Oxytricha can be grown in 50-liter fermentation vats. The cultures grow to a uniform density of about 6000 cells/ml.

Notes: Ichthyophthirius multifiliis, a parasitic ciliate of freshwater fishes, was found to have surface antigens (Ag) which elieited immobilizing antibodies (Ab) when injected into rabbits. An effort was made to purify and characterize these Ag (referred to as immobilization Ag) because of their potential role in protective immunity in fishes. Mice immunized with theront cilia were used for production of immobilizing monoclonal antibodies (MAb). Hybridomas were screened by indirect immunofluorescent light microscopy and immobilization of live parasites. Six hybridomas producing immobilizing MAb were cloned. Immobilizing MAb were used to affinity purify Ag solubilized with Triton X-114 and Na deoxycholate. Two membrane protein Ag of approximately 48 and 60 kDa were identified. Immobilizing MAb failed to react with these Ag on Western blots and, conversely, MAb that reacted with the Ag on Western blots did not immobilize live organisms. These results suggest that immobilization required native conformational epitopes which were altered by Western blotting procedures. Individual MAb reactive on Western blots recognized both the 48- and 60-kDa proteins indicating the presence of common epitopes. Affinity purified Ag elicited immobilizing antisera when injected into rabbits, mice, and channel catfish.

Notes: . The paper is concerned with the principles upon which coccidia of the genus Eimeria may be characterized. Reference strains for comparative purposes usually are not available and the limitations of morphological data for speciation are discussed. The value of other parameters are considered such as host and site specificity, pathogenicity, immunological specificity, pre-patent period, sporulation time, enzyme variation, and DNA buoyant density. The weight afforded to each of these parameters for specific identification may vary according to the parasite and host studied. Determinations of physiological and behavioral characteristics that are now becoming available should be included in species definitions wherever possible.

Notes: . Changes in nuclei and nucleoli of cells of chicken cecum infected with Eimeria tenella were studied in living cells by interference microscopy and in fixed and stained tissues using light level microscopy. As soon as merozoites began to transform into second generation meronts, there was an increase in the size of both the nucleus and the nucleolus of the host cell. The dry weight of the nucleus increased somewhat, but there was a greater increase and a correlation of the dry mass of the nucleolus with the size of the parasite as measured by interference microscopy. In fixed and stained tissues, there was a correlation between the area of the nucleolus and the area of the parasite. Removal of nucleic acids with DNase and/or RNase showed high concentrations of both in the nucleoli and a residue of protein. The increased nucleolar size indicates a high level of transcription in infected cells and allows the conclusion that the parasite somehow induces transcription to occur. Since transcription is a highly specific process, the high degree of host and site specificity shown by nearly all coccidia is consistent with a hypothesis that the coccidia share a portion of the host genome.

Notes: A recent isolate of Eimeria praecox, strain G, was obtained from Georgia and purified. Studies of the life history, pathogenicity, and cross-immunity of the isolate were conducted to verify its identity. In inoculated three-week-old chickens, the occurrence of merogony and gametogony was limited to the superficial epithelium of the upper intestine. Oocysts, 23 × 19.5 äm, with a shape index of 1.17 were first observed 83 h after inoculation. Mortality and morbidity were not observed in any of the experimental birds. However, there was a positive correlation between dose of oocysts, reduced weight gain, and the incidence of exudative diathesis. These studies showed that E. praecox depresses weight gains in chickens and may be of economic importance. Although complete immunity to avian coccidiosis is believed to be species specific, chickens immune to E. praecox (G) or E. acervulina had a degree of cross-immunity to a heterologous challenge. Electrophoretic analysis of glucose phosphate isomerase and lactate dehydrogenase prepared from the European strain of E. praecox and E. praecox (G) showed no differences, confirming the identity of the isolate as E. praecox.

Notes: A field plot experiment was carried out on an established grassland sward from 1983–88 inclusive to examine the effects of time of application, chemical form of nitrogen (N) and cattle slurry dry matter (DM) content on yield and efficiency of N use. Four forms of fertilizer N (a semi-organic fertilizer, a combined 2.1:1 (w/w) semi-organic/calcium ammonium nitrate (CAN) fertilizer, CAN and urea, each supplying 300 kg N ha−1 year−1, were applied with or without unseparated or separated cattle slurry at 93 and 73 g kg−1 DM respectively, both supplying approximately 150 kg N ha−1 year−1. All fertilizers and slurries were applied in three equal dressings (February/March, May/June and July/August). The efficiency of use of fertilizer and slurry N was evaluated by measuring DM yield, N uptake and apparent recovery of N in herbage at all harvests during each growing season.Fertilizer N application significantly increased (P〈0.001) the mean yields of herbage at each harvest in all years. The form of fertilizer N had no significant effect (P〉0.05) on first harvest and total herbage yields, nor on N uptakes by herbage at the first harvest in any year. The performance of urea and of CAN was more variable at the second and third harvests relative to that of the semi-organic or combined 2.2:1 (w/w) semi-organic/CAN sources which had similar efficiencies of N use. Lower DM production was associated with reduced uptake of N. Values for mean overall apparent recovery of N ranged from 57.9 ± 2.67% for the semi-organic fertilizer to 50.2±3.05% for CAN.Unseparated cattle slurry and separated cattle slurry produced similar herbage yields and N responses that were lower and more variable than with fertilizer N. The overall mean apparent recovery of N from unseparated cattle slurry was 25.5 ± 5.03% compared to 5.0 ± 4.74% for separated cattle slurry. Efficiency of N use was highest with spring applications and least with mid-season applications. Recoveries ranged from −29% for separated cattle slurry applied in June 1984 to 56% for unseparated and separated cattle slurry applied in February 1988 and June 1987 respectively. No interactions were recorded between cattle slurry and fertilizer N in terms of DM production or N uptake by herbage.The results of this study support the use of a fertilizer N source, selected on a least-cost basis, in combination with slurry to promote spring herbage production. For subsequent production, N should be supplied in fertilizer form only. The use of urea is risky under low rainfall conditions. Mechanical separation did not improve the efficiency of use of slurry N.

Notes: Hydrogen gas was discovered within the steel casing above standing water in a percussion-drilled borehole on the Hanlord Site in south-central Washington state. In situ measurements of the borehole fluids indicated anoxic, low-Eh (〈-400 mV) conditions. Ground water sampled from adjacent wells in the same formation indicated that the ground water was oxygenated. H2 was generated during percussion drilling, due to the decomposition of borehole waters as a result of aqueous reactions with drilled sediment and steel from the drilling tools or casing. The generation of H2 within percussion-drilled boreholes that extend below the water table may be more common than previously realized. The ambient concentration of H2 produced during drilling was limited by microbial activity within the casing-resident fluids. H2 was generated abiotically in the laboratory, whereby sterilized borehole slurry samples produced 100 times more H2 than unsterilizcd samples. It appears that H2 is metabolized by microorganisms and concentrations might be significantly greater if not for microbial metabolism.