Susceptibilities against bovine lactoferrin with microorganisms isolated from mastitic milk

Antibacterial effects of bovine lactoferrin were studied in vitro against microorganisms isolated from mastitic milk in Tokachi area, Hokkaido, Japan. Microorganisms isolated were Escherichia coli (11 isolates), Klebsiella pneumoniae (5 isolates), enterococci (8 isolates), Staphylococcus aureus (10 isolates), coagulase negative staphylococci (CNS, 13 isolates), streptococci (11 isolates), Prototheca zopfii (7 isolates) and yeast-like fungi (9 isolates). Lactoferrin has been known as a multifunctional protein and its antimicrobial effect is one of the most essential function of it. In order to compare their susceptibilities against lactoferrin, the minimal inhibitory concentration values were estimated by a microplate assay method using 96-well microplate, which involved measuring the optical density of the cultures. Prototheca zopfii was highly sensitive to bovine lactoferrin and complete inhibition of this microorganism was observed even at the low concentration of 7 mug/ml. On the other hand, E. coli and enterococci showed resistance against lactoferrin action and staphylococci showed strain-dependent resistance.     

Commercial harvest of hatchery-reared masu salmon Oncorhynchus masou estimated by a coast-wide sampling program in Hokkaido, northern Japan, and the two-stage sampling schemes of landings

SUMMARY: This paper evaluates the stocking effectiveness of masu salmon Oncorhynchus masou in Hokkaido, northern Japan, through a coast-wide two-stage sampling survey of commercial landings. From January to June 1994–1996, commercial landings of masu salmon at 33–36 fish markets were sampled at 7–10 days intervals, and 60 866–72 124 fish were examined for marks indicated by fin clips. Based on the survey data, numbers of total and hatchery-reared masu salmon landed were estimated. To examine the structure of the errors, stratification of fish markets was implemented on the basis of geography and magnitudes of landings, and the stratification improved accuracy and precision of the estimates. Accuracy of the estimated numbers of total fish was evaluated by being compared to the true numbers of masu salmon landings reported by fishermen's cooperative associations. Estimates of total masu salmon landings were within ± 10% of the true numbers. The estimated recovery rates (± SE) for hatchery-reared masu salmon smolts were variable ranging from 0.18 (± 0.06) to 3.50 (± 0.41)% among the stocked groups. An optimal sampling strategy was examined to obtain precise estimates for future studies.     

The detection of bovine lactoferrin binding protein on Trypanosoma brucei

Trypanosoma brucei, the causative agent of sleeping sickness in humans, requires transferrin (TF) for growth. Therefore, T. brucei has a TF receptor that allows it to obtain iron from TF. Lactoferrin (LF), a member of the TF family protein, is an iron-binding protein that is found in most biological fluids of mammals. LF has been shown to interact with some bacteria species by specific receptor-ligand binding. We examined the ability of T. brucei to bind bovine LF (bLF) by using a fluorescence test, streptavidin-biotin (SAB) microplate analysis, and far Western blotting using a biotin-streptavidin system. We found that bLF bound to components of T. brucei, and that bLF hydrolysate disrupted the sites responsible for binding to parasite proteins. Furthermore, bLF, human LF, bovine TF, and ovotransferrin bound same proteins of T. brucei, which exhibited molecular masses of 40 and 43 kDa. The N-terminal amino acid sequence of the 40 kDa bLF binding protein was identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH).