Freeze-thaw-induced lateral transfer of non-conjugative plasmids by in situ transformation in <Emphasis Type="Italic">Escherichia coli</Emphasis> in natural waters and food extracts

Freeze-thaw treatment of condensed suspensions of mixed Escherichia coli strains in natural waters and food extracts caused in situ lateral transfer of non-conjugative plasmids. This phenomenon also occurred in distilled water and LB broth, and after 1–2 months of preservation at −20°C. The sensitivity of lateral transfer towards DNase activity suggested the involvement of in situ transformation. There were no clear correlations between transformation frequency and the chemical characteristics (Ca²⁺, Mg²⁺, and pH) of the samples. These results suggest the possibility that freeze–thaw-induced lateral gene transfer between bacterial cells occurs in natural and human environments.     

Larval competition for food between wild and hatchery-reared ayu, Plecoglossm altivelis Temminck et Schlegel, in culture ponds

To examine the larval competition between wild and hatchery ayu in the culture ponds, mixed rearing of 580 000 wild and 520 000 hatchery larvae was carried out in two 25-m³ ponds for 3 months, in contrast to the control in which 860 000 wild larvae were reared in another pond.
The number of wild larvae in the mixed-rearing treatments decreased rapidly 20 days after the start of mixed rearing, in contrast to hatchery larvae. Mortality of wild larvae was almost 100% at the end of the experiment (3 months), while the hatchery larvae showed the usual survival rate of 15–16%. In the control pond, however, 16% of the wild larvae survived. The wild larvae grew much slower (0.10mmday ^-1) than the hatchery larvae (0·26 mm day ^-1) in the mixed-rearing ponds, whereas the wild larvae in the control pond showed almost the same growth rate (0·24 mm day ^-1) as hatchery larvae. On day 6 the gut fullness of wild larvae was only 30% of that of the hatchery larvae in the mixed-rearing experiments. On day 46 the wild larvae occurred deeper in the mixed-rearing ponds than the hatchery larvae. This depth difference in vertical distribution appeared to cause a disadvantage for the wild larvae in the competition with hatchery larvae, since the food was supplied at the surface. Thus, the wild larvae starved and died.     

Domestic dog demographics and estimates of canine vaccination coverage in a rural area of Zambia for the elimination of rabies

BackgroundAn estimated 75% or more of the human rabies cases in Africa occur in rural settings, which underscores the importance of rabies control in these areas. Understanding dog demographics can help design strategies for rabies control and plan and conduct canine mass vaccination campaigns effectively in African countries.Methodology/Principal findingsA cross-sectional survey was conducted to investigate domestic dog demographics in Kalambabakali, in the rural Mazabuka District of Zambia. The population of ownerless dogs and the total achievable vaccination coverage among the total dog population was estimated using the capture-recapture-based Bayesian model by conducting a canine mass vaccination campaign. This study revealed that 29% of the domestic dog population was under one year old, and 57.7% of those were under three months old and thus were not eligible for the canine rabies vaccination in Zambia. The population growth was estimated at 15% per annum based on the cross-sectional household survey. The population of ownerless dogs was estimated to be small, with an ownerless-to-owned-dog ratio of 0.01–0.06 in the target zones. The achieved overall vaccination coverage from the first mass vaccination was estimated 19.8–51.6%. This low coverage was principally attributed to the owners’ lack of information, unavailability, and dog-handling difficulties. The follow-up mass vaccination campaign achieved an overall coverage of 54.8–76.2%.Conclusions/SignificanceThis paper indicates the potential for controlling canine rabies through mass vaccination in rural Zambia. Rabies education and responsible dog ownership are required to achieve high and sustainable vaccination coverage. Our findings also propose including puppies below three months old in the target population for rabies vaccination and emphasize that securing an annual enforcement of canine mass vaccination that reaches 70% coverage in the dog population is necessary to maintain protective herd immunity.     

Efficient synthesis of polypeptide‐α‐thioester by the method combining polypeptide expression and chemical activation for the semi‐synthesis of interferon‐γ having oligosaccharides

In order to synthesize interferon‐γ glycoform having an oligosaccharide at the 97 position by a semi‐synthetic method, interferon‐γ‐polypeptide‐(1–94)‐α‐hydrazide was prepared by the specific Cys‐cyanylation of polypeptide‐(1–94)‐Cys‐His₆ expressed from E. coli and subsequent hydrazinolysis in 22% yield (two steps). This polypeptide‐α‐hydrazide was then converted into corresponding polypeptide‐α‐thioester under NaNO₂/acid conditions followed by thiolysis in 83% yield. Copyright © 2014 European Peptide Society and John Wiley & Sons, Ltd.     

Fructose-diphosphate aldolase of Horseshoe crab (Tachypleus tridentatus).

Fructose-diphosphate aldolase [ED 4.1.2.13] was isolated from horseshoe crab ( living fossil) muscle and some molecular and enzymatic properties were examined. The enzyme was a tetramer with a molecular weight of about 160,000. The enzyme activity was inhibited by reduction with borohydride in the presence of the substrate and was inactivated by carboxypeptidase A [EC 3.4.12.2] digestion. The pH optima for fructose-diphosphate (FDP) and fructose-1-phosphate (F1P) activities were 6.5--8 and 7.5--8.2, respectively. The ratio of FDP/F1P activities was 30 and Km values were 1.7 times 10- minus 5 M and 2.5 times 10- minus 3 M, respectively, for the two substrates. The horseshoe crab aldolase was classified as class 1, type A, based on the results obtained. Extensive homology in various properties of the enzyme was observed when it was compared with enzymes from other sources, though some differences could be found in the amino acid composition and in the kinetic properties.     

Field assessment of B-2 as a new molluscicide for the control of Oncomelania nosophora

Sodium 2,5-dichloro-4-bromophenol named B-2 was evaluated for application in the field as a molluscicde against Oncomelania nosophora. A dosage of 10 g/m2 as 25% liquid form or 25 g/m2 as 10% granular form of B-2 was suggested as a standard dose for the control of O. nosophora. The residual concentration of B-2 in the soil after application was observed to decrease more rapidly than did that of NaPCP. The residual level of B-2 in the rice grains harvested from the paddy field in which the chemical was applied at most 4 times during the planting season did not exceed 0.03 ppm.     

Establishment of a novel embryonic stem cell line by a modified procedure

To generate mutant mice, embryonic stem (ES) cells are used as a vehicle for introducing mutations. The establishment of ES cells is diffucult because it requires specific skills and it is time-consuming. We established a novel ES cell line derived from hybrid mice between C57BL/6 and DBA/2 using a modified method. To collect a large number of preimplantational embryos, we collected embryos at the 8-cell stage and cultured them to blastocysts, whereas the usual procedure of preparing the delayed blastocysts demands technical skills. To eliminate unnecessary female cells at an initial stage of inner cell mass culture, male clones were selected by polymerase chain reaction to detect the mouseSry gene. The established ES cell line efficiently contributed to the germ-line when injected into 8-cell embryos of ICR mice. This potency was maintained after manipulation throughout gene targeting.Abbreviations DMEM Dulbecco's modified Eagle's medium - FBS fetal bovine serum - FIAU 1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-iodouracil - LIF leukemia inhibitory factor - NEAA non-essential amino acids