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Kanae SHIOKAWA Chandika D. GAMAGE Nobuo KOIZUMI Yoshihiro SAKODA Kenta SHIMIZU Yoshimi TSUDA Kumiko YOSHIMATSU Jiro ARIKAWA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2016,78(2):221-230
The applicability of the recombinant LipL32 for serodiagnosis of leptospiral infection in field rodents was
assessed in this study. An immunodominant region of LipL32 was determined by monoclonal antibodies, and then,
truncated LipL32 (tLipL32) was designed to contain the region (87–188th amino acid). The tLipL32 was compared
between two recombinant expression hosts Escherichia coli and Pichia
pastoris in ELISA. With field rat sera, tLipL32 expressed by P. pastoris
(tLipL32p) had high antigenicity without background reactions, while tLipL32 expressed by E.
coli (tLipL32e) showed high background reactions, which were reduced by pre-adsorption of sera with
E. coli. To evaluate tLipL32-ELISA, field rat sera were tentatively divided into a
Leptospira infection positive (12 sera) and a negative group (12 sera) based on the results
from flaB gene PCR of kidney samples and WB with whole Leptospira cell.
Consequently, the sensitivity of tLipL32p-ELISA for field rat sera was 83% . A similar result was obtained
from tLipL32e-ELISA with adsorbed sera, (92%). However, sensitivity of tLipL32e-ELISA using sera without an
adsorption treatment was 50%. Regardless of the expression host, tLipL32-ELISA had 100% specificity and
sensitivity in experimentally infected laboratory rats. These results suggest that recombinant LipL32
expressed by P. pastoris is more applicable for serodiagnosis in field rats due to a lack of
background reaction. 相似文献
3.
Akihiro Okamura Yoshiaki Yamada Noriyuki Horie Tomoko Utoh Naomi Mikawa Satoru Tanaka Katsumi Tsukamoto 《Fisheries Science》2008,74(3):642-648
ABSTRACT: The effects of silvering state of wild female Japanese eels Anguilla japonica on the success of induced maturation and the following spawning were examined. Thirty-eight females, collected in Mikawa Bay, were divided into four stages based on their silvering state: yellow (Y1), late-yellow (Y2), silver (S1) and late silver eels (S2). Despite injections of salmon pituitary extract (SPE) through the standard technique, Y1 and Y2 eels did not respond to the treatment with undeveloped gonad (gonad-somatic index [GSI]: 0.3–0.9), and all these females died by 5 weeks, probably due to an abnormal physiological condition. Most S1 (81%) and S2 eels (100%) matured completely (GSI: 17.8–51.4), and finally spawned successfully (69% for S1, 89% for S2). S2 eels fully matured with oocytes of over 750 μm in diameter by significantly smaller number of injections of SPE (5–6 times) than the case of S1 eels (6–8 times). The amount of eggs released by S2 eels (0.65 ± 0.11 g/fish per body weight [BW]) was significantly larger than those by S1 eels (0.54 ± 0.09 g/fish per BW). There was no difference in fertilization and hatching rates between eggs released by S1 eels and those of S2 eels. These results indicate that the success of induced maturation and spawning in wild female Japanese eels depends on their silvering state, and matured eggs can be obtained efficiently through the use of S2 eels rather than other stages. 相似文献
4.
Paddy fields converted into winter wheat fields in Hokkaido, Japan, receive extremely high snowfall, creating a risk of flood damage to crops in spring due to waterlogging of snowmelt runoff and poor drainage. Meanwhile, in June there is relatively little rainfall, and a lack of moisture inhibits winter wheat growth. Therefore, we developed a method involving a series of 30-cm-deep ditches in agricultural fields to be used for drainage during the flood-prone period and for furrow irrigation during the dry period using water drawn from the canals that feed the paddy fields. The ditches are called ‘hybrid ditches’ as they are able to perform both drainage and irrigation functions. In this study, we investigated the optimal construction timing and spacing for hybrid ditches. We also evaluated their ability to improve the drainage and irrigation of winter wheat. We found that the optimal timing for digging hybrid ditches is immediately after sowing, and the inter-ditch spacing for irrigation should be 15 m or less. The hybrid ditches promoted increased soil temperature and healthy development of wheat plants by improving drainage during the flood-prone period. In addition, water was successfully supplied via the hybrid ditches to irrigate the fields in June. Under experimental conditions in which rainfall was excluded, grain yield was 10% higher and percent protein content was more than 1% point greater in the irrigated plot compared with the non-irrigated plot. Grain yield was also observed to increase by 3–29% in demonstration tests conducted at local farms. From these results, we conclude that hybrid ditches are capable of improving the growth and yield of winter wheat by improving drainage and providing irrigation in converted paddy fields in Hokkaido. 相似文献
5.
ABSTRACT: Hermatypic (reef-building) corals harbor dinoflagellate endo-symbionts Symbiodinium spp. In studying gene expression in such symbiotic corals, problems arise regarding how to distinguish the coral and symbiont mRNA, and how to estimate their fractions in the mRNA population of the holobiont (symbiotic complex of the coral and Symbiodinium cells). In this study, these issues were addressed using juveniles of hermatypic coral Acropora tenuis in symbiosis with Symbiodinium cells of strain PL-TS-1. First, the guanine-cytosine (GC) contents were determined in expressed sequence tags (EST) from PL-TS-1 cells cultured in vitro and symbiont-free larvae of A. tenuis , and their average GC contents were found to be significantly different. The average GC content of the EST from the holobiont was much closer to that of A. tenuis larvae, suggesting that the majority (>90%) of mRNA isolated from the holobiont originated in the host. In protein-coding sequences, little overlap was observed between the GC-content distributions of PL-TS-1 cells and A. tenuis larvae. All of the coding sequences ( n = 59) found in the A. tenuis EST had GC contents below 0.5, whereas the GC content exceeded 0.5 in the majority (43/44) of coding sequences from the nuclear genome of PL-TS-1 cells. 相似文献
6.
Interannual variations in abundance, timing of outmigration from rivers, growth rate and condition of juvenile chum salmon
(Oncorhynchus keta) were studied in the Nemuro Strait (eastern Hokkaido, Japan) during 1999–2002 to establish a possible relationship to zooplankton
abundance. The otolith microstructure of juveniles was examined each year in late June to determine their time and size at
sea entry (i.e., outmigration), and to estimate the early marine growth rates. Salmon outmigration peaked in mid- or late
May, which coincided, in three of the four study years, with the peak release of juveniles into rivers within the study area.
Abundance, growth rate and condition of fish were higher in 2001, when—compared to other years—smaller fish experienced higher
growth rates, coinciding with greater zooplankton abundance for that year. Our results suggest that high zooplankton abundance
positively influenced juvenile chum salmon growth and the condition of the fish during their early marine life despite their
small size at sea entry. 相似文献
7.
Antimicrobial Activity of Culture Filtrate of Bacillus amyloliquefaciens RC-2 Isolated from Mulberry Leaves 总被引:1,自引:0,他引:1
ABSTRACT A potential antagonist, Bacillus amyloliquefaciens strain RC-2, against Colletotrichum dematium, mulberry anthracnose fungus, was obtained from healthy mulberry leaves by in vitro and in vivo screening techniques. Application of culture filtrate of RC-2 inhibited disease on mulberry leaves, indicating that suppression was due to antifungal compounds in the filtrate. Development of mulberry anthracnose on mulberry leaves was inhibited only when the culture filtrate was applied before fungal inoculation, and it was not inhibited by application after inoculation. These results suggest that the antifungal compounds in the filtrate exhibit a preventive effect on the disease. Peptone significantly increased production of the antifungal compounds. The culture filtrate of RC-2 also inhibited the growth of several other phytopathogenic fungi and bacteria, such as Rosellinia necatrix, Pyricularia oryzae, Agrobacterium tumefaciens, and Xanthomonas campestris pv. campestris, in vitro. From the culture filtrate of RC-2, seven kinds of antifungal compounds were isolated by high performance liquid chromatography analysis, and one of the compounds was determined as iturin A2, a cyclic peptide, by nuclear magnetic resonance and fast atom bombardment mass analysis. 相似文献
8.
9.
Nurun Nahar Khanam Junichi Kihara Yuichi Honda Toshihide Tsukamoto Sakae Arase 《Journal of General Plant Pathology》2005,71(4):285-288
When detached broad bean leaves were preinoculated with virulent strain B304 of Botrytis cinerea 24 h before a challenge inoculation with strain B304, lesion formation by B304 was significantly inhibited in red light but not in the dark. In leaves that were preinoculated with avirulent strain 021 and then challenged by B304, however, lesion formation was not inhibited even under red light. Such differences in lesion formation after the challenge inoculation with an avirulent strain were also observed with lesions caused by Alternaria alternata, a nonpathogen of broad bean and by avirulent strain 021R in the presence of germination fluid from spores of strains B304 and 021R. These results suggest the possibility that virulent B. cinerea produced a suppressor involved in induced susceptibility and an elicitor involved in resistance induced by red light during spore germination. 相似文献
10.
Chicken anaemia virus (CAV) infectivity and the effect of highly virulent infectious bursal disease virus (hv IBDV) infection on CAV's infectivity were examined in chickens inoculated with CAV or inoculated dually with CAV and hv IBDV. Five chickens inoculated dually with hv IBDV at 35 days old and then with CAV at 40 days old exhibited no clinical signs of disease, but showed atrophic bursae of Fabricius when necropsied 4 weeks later. Upon examining the chickens at 7 days postinoculation (dpi) with CAV, it was found that hv IBDV infection had inhibited production of virus neutralising (VN) antibody to CAV, and that it was possible to recover CAV from plasma of these chickens. Although VN antibody to CAV appeared after 14 dpi, CAV was recovered from blood cells (BC s) at high titres ranging from 10(2.5)to 10(5.5)TCID(50)/0.1 ml, 7 to 28 dpi in IBDV -induced immunosuppressed chickens. In addition, CAV was sporadically recovered, using rectal swabs, from the dually inoculated chickens at low titers, ranging from 10(1.0)to 10(2. 0)TCID(50)/0.1 ml). In contrast, although CAV was recovered from BC s in most of the chickens inoculated with CAV alone, the titers were lower (10(1.0)to 10(2.5)TCID(50)/0.1 ml). No CAV was detected from the rectal swabs of these chickens. The results of virus recovery were confirmed by polymerase chain reaction. This study first examined the persistency of CAV in BC s and the effective enhancement of primary CAV infection as a result of immunosuppression caused by hv IBDV infection. 相似文献