Integrating cultural, economic and environmental requirements for fish production in Borno State

The study examines the integration of cultural, economic and environmental requirements for fish production in Borno State, Nigeria. A reconnaissance survey was conducted transferring some selected Local Government Areas. 60 questionnaires were administered in the six Local Governments representing Southern Borno State with Biu and Shani, central Borno with Konduga & Jere and Northern Borno with Gubia and Kukawa respectively. There is no cultural constraint to fish production but about 63% prefers to invest in other farming activities than in fish farming. 33% are not aware that fish can be cultured apart from getting it from the wild. 35% have the impression that fish farming ventures can be handled by government only. The economic earnings for fish production are high especially in some parts of Northern Borno, but the Local market potentials throughout the state are great. Nigeria has suitable soil for ponds apart from few locations at the central and Northern Borno that are made by sandy soil. Numerous perennial and seasonal rivers, streams, lakes, pools and flood plains adequate for fish culture especially in Southern Borno exist. The mean annual rainfall can result in some water storage in ponds. In areas where the annual precipitation is less than 550mm, exist few flow boreholes with potentials for fish production. The temperature regime may support growth and survival of fish even during the hottest months of the year (March, April and May). With the understanding and manipulation of these requirements, fish production in Nigeria can be greatly enhanced

Structural requirements for catalysis: studies on RTEM-1 β-lactamase

β-lactamases are a group of enzymes that confer resistance to penam and cephem antibiotics by hydrolysis of the β-lactam ring, thereby inactivating the antibiotic. Crystallographic and computer modeling studies of RTEM-1 β-lactamase have indicated that Asp 132, a strictly conserved residue among the class A β-lactamases, appears to be involved in substrate binding, catalysis, or both. To study the contribution of residue 132 to β-lactamase function, site saturation mutagenesis was used to generate mutants coding for all 20 amino acids at position 132. Phenotypic screening of all mutants indicated that position 132 is very sensitive to amino acid changes, with only N132C, N132D, N132E, and N132Q showing any appreciable activity. Kinetic analysis of three of these mutants showed increases in K_M, along with substantial decreases in k_(cat). Efforts to trap a stable acyl-enzyme intermediate were unsuccessfuL These results indicate that residue 132 is involved in substrate binding, as well as catalysis, and supports the involvement of this residue in acylation as suggested by Strynadka et al.

Crystallographic and computer modeling studies of RTEM-1 β-lactamase have indicated that Lys 73 and Glu 166, two strictly conserved residues among the class A β-lactamases, appear to be involved in substrate binding, catalysis, or both. To study the contribution of these residues to β-lactamase function, site saturation mutagenesis was used to generate mutants coding for all 20 amino acids at positions 73 and 166. Then all 400 possible combinations of mutants were created by combinatorial mutagenesis. The colonies harboring the mutants were screened for growth in the presence of ampicillin. The competent colonys' DNA were sequenced, and kinetic parameters investigated. It was found that lysine is essential at position 73, and that position 166 only tolerated fairly conservative changes (Aspartic acid, Histidine, and Tyrosine). These functional mutants exhibited decreased kcat's, but K_M was close to wild-type levels. The results of the combinatorial mutagenesis experiments indicate that Lysis absolutely required for activity at position 73; no mutation at residue 166 can compensate for loss of the long side chain amine. The active mutants found--K73K/E166D, K73KIE166H, and K73KIE166Y were studied by kinetic analysis. These results reaffirmed the function of residue 166 as important in catalysis, specifically deacylation.

The identity of the residue responsible for enhancing the active site serine (Ser 70) in RTEM-1 β-lactamase has been disputed for some time. Recently, analysis of a crystal structure of RTEM-1 β-lactamase with covalently bound intermediate was published, and it was suggested that Lys 73, a strictly conserved residue among the class A β-lactamases, was acting as a general base, activating Ser 70. For this to be possible, the pK_a of Lys 73 would have to be depressed significantly. In an attempt to assay the pK_a of Lys 73, the mutation K73C was made. This mutant protein can be reacted with 2-bromoethylamine, and activity is restored to near wild type levels. ^(15)N-2-bromoethylamine hydrobromide and ^(13)C-2-bromoethylamine hydrobromide were synthesized. Reacting these compounds with the K73C mutant gives stable isotopic enrichment at residue 73 in the form of aminoethylcysteine, a lysine homologue. The pK_a of an amine can be determined by NMR titration, following the change in chemical shift of either the ^(15)N-amine nuclei or adjacent Be nuclei as pH is changed. Unfortunately, low protein solubility, along with probable label scrambling in the Be experiment, did not permit direct observation of either the ^(15)N or ^(13)C signals. Indirect detection experiments were used to observe the protons bonded directly to the ^(13)C atoms. Two NMR signals were seen, and their chemical shift change with pH variation was noted. The peak which was determined to correspond to the aminoethylcysteine residue shifted from 3.2 ppm down to 2.8 ppm over a pH range of 6.6 to 12.5. The pK_a of the amine at position 73 was determined to be ~10. This indicates that residue 73 does not function as a general base in the acylation step of the reaction. However the experimental measurement takes place in the absence of substrate. Since the enzyme undergoes conformational changes upon substrate binding, the measured pK_a of the free enzyme may not correspond to the pK_a of the enzyme substrate complex.

Species Profiles: Life Histories and Environmental Requirements of Coastal Fishes and Invertebrates (Pacific Northwest): Amphipods

This report wi11 focus largely on the suborders Gammaridea, Caprellidea, and Hyperiidea because of their importance in coastal areas of the northeast Pacific Ocean. (PDF contains 27 pages)

Species Profiles: Life Histories and Environmental Requirements of Coastal Fishes and Invertebrates (Pacific Southwest): Amphipods

(PDF contains 24 pages)

Species Profiles: Life Histories and Environmental Requirements of Coastal Fishes and Invertebrates (Mid-Atlantic): Bay anchovy

The bay anchovy occurs along the Atlantic and Gulf of Mexico coasts, from Cape Cod, Massachusetts, to Yucatan, Mexico (Hildebrand 1963), except for the Florida Keys where it is apparently absent (Daly 1970). (PDF contains 22 pages)

Species Profiles: Life Histories and Environmental Requirements of Coastal Fishes and Invertebrates (Gulf of Mexico): Bay anchovy and Striped Anchovy

(PDF contains 24 pages)

Species Profiles: Life Histories and Environmental Requirements of Coastal Fishes and Invertebrates (Pacific Southwest): Northern anchovy

Three genetically distinct groups: British Columbia to northern California, Southern California to the northern Baja peninsula, and central and southern Baja California. (PDF contains 21 pages)