RESUMO
Korean rose bitterling (Rhodeus uyekii) is a freshwater fish endemic to Korea. Natural populations of this species have experienced severe declines as a result of habitat fragmentation and water pollution. To conserve and restore R. uyekii, the genetic diversity of this species needs to be assessed at the population level. Eighteen novel polymorphic microsatellite loci for R. uyekii were developed using an enriched partial genomic library. Polymorphisms at these loci were studied in 150 individuals collected from three populations. The number of alleles at each locus ranged from 3 to 47 (mean = 17.1). Within the populations, the observed heterozygosity ranged from 0.032 to 1.000, expected heterozygosity from 0.082 to 0.967, and polymorphism information content from 0.078 to 0.950. Six loci showed significant deviation from Hardy-Weinberg equilibrium after Bonferroni's correction, and no significant linkage disequilibrium was detected between most locus pairs, except in three cases. These highly informative microsatellite markers should be useful for genetic population structure analyses of R. uyekii.
Assuntos
Peixes/genética , Biblioteca Genômica , Repetições de Microssatélites , Alelos , Animais , Genótipo , Polimorfismo GenéticoRESUMO
Korean starry flounder, Platichthys stellatus (Pleuronectidae), is one of the most economically important fishery resources in Korea. We investigated the effect of current artificial reproduction in a hatchery facility, genetic divergence between the broodstock and their offspring populations of starry flounder in a hatchery strain to be stocked into natural sea areas was accessed using 9 polymorphic nuclear microsatellite DNA loci. High levels of polymorphism were observed between the 2 populations. A total of 96 alleles were detected at the loci, with some alleles being unique in the broodstock. Allelic variability ranged from 8 to 17 in the broodstock and from 7 to 12 in the offspring population. Average observed and expected heterozygosities were estimated at 0.565 and 0.741 in the broodstock samples and 0.629 and 0.698 in the offspring population, respectively. Although no statistically significant reductions were found in heterozygosity or allelic diversity in the offspring population, a considerable loss of rare alleles was observed in the offspring population compared with that in the broodstock. Significant genetic difference was detected between the broodstock and offspring populations (FST = 0.021, P < 0.05). These results suggest that more intensive breeding practices for stock enhancement might have resulted in a further decrease of genetic diversity. Thus, genetic variations of broodstock and progeny should ideally be monitored in both breeding and release programs as a routine hatchery operation in order to improve the starry flounder hatchery management. This information might be useful for fishery management and aquaculture industry of P. stellatus.
Assuntos
Linguado/genética , Repetições de Microssatélites , Polimorfismo Genético , Animais , Cruzamento , Pesqueiros , Frequência do Gene , Loci Gênicos , Genética Populacional , República da CoreiaRESUMO
By using IL-5 transgenic mice, it has been shown that eosinophils might play a key role in elimination of larval stages of nematode infections. The present study was carried out to clarify molecular mechanisms involved in the eosinophil-mediated killing of Nippostrongylus brasiliensis larvae. The larvicidal activity was observed in the presence of normal serum in vitro. Electron microscopic observations revealed firm attachment of eosinophils to the cuticular surface of larvae, which was damaged by electron-dense materials released from eosinophils. The larvicidal activity was abrogated by heat- or zymosan-treatment of the serum, whereas depletion of IgG or IgM from the serum did not interfere with eosinophil adhesion and killing. Moreover, pretreatment of eosinophils with monoclonal antibodies against CD11b or VLA-4 inhibited the eosinophil-mediated killing of larvae. Immunofluorescent staining demonstrated the deposition of C3c and plasma fibronectin on the cuticle of the larvae. These results indicate that interactions between CD11b and VLA-4 and their respective counter-ligands deposited on the cuticle are essential in eosinophil-mediated adhesion and damage to larvae of N. brasiliensis.
Assuntos
Proteínas do Sistema Complemento/fisiologia , Eosinófilos/imunologia , Fibronectinas/metabolismo , Nippostrongylus/imunologia , Infecções por Strongylida/imunologia , Animais , Adesão Celular , Integrina alfa4beta1 , Integrinas/metabolismo , Interleucina-5/genética , Larva/imunologia , Antígeno de Macrófago 1/metabolismo , Camundongos , Camundongos Endogâmicos C3H , Camundongos Transgênicos , Nippostrongylus/crescimento & desenvolvimento , Receptores de Retorno de Linfócitos/metabolismo , Infecções por Strongylida/parasitologiaRESUMO
Eosinophil and IgE responses of interleukin (IL)-5 transgenic and normal C3H/HeN mice were studied after experimental infection with Nippostrongylus brasiliensis (Nb). Intestinal worms were recovered at day 5 post-infection (PI), and numbers of total white blood cells (WBC) and eosinophils, and total serum IgE and anti-hapten (dinitrophenyl) (DNP) specific IgE titers, were measured at days 0, 14 and 21 PI. IL-5 mice appeared resistant to Nb infection showing a significantly lower worm recovery rate than normal mice (P < 0.05). Total WBC and eosinophil counts (/mm3) were significantly increased in Nb infected normal mice (P < 0.05), but unchanged (total WBC) or decreased (eosinophils) in IL-5 mice at day 21 PI. The total serum IgE level remarkably increased in normal mice, but only a little in IL-5 mice at days 14 and 21 PI. Priming with DNP brought about more remarkable increases of the total and anti-DNP specific IgE in normal mice than in IL-5 mice. The results show that IL-5 mice are resistant to Nb infection, and that eosinophil and IgE responses in these mice are not augmented by Nb infection.
Assuntos
Anticorpos Anti-Helmínticos/sangue , Eosinófilos/imunologia , Imunoglobulina E/sangue , Interleucina-5/genética , Nippostrongylus , Infecções por Strongylida/imunologia , Animais , Feminino , Contagem de Leucócitos , Camundongos , Camundongos Endogâmicos C3H , Camundongos Transgênicos/imunologia , Nippostrongylus/imunologiaRESUMO
Nippostrongylus brasiliensis infection is characterized by blood and tissue eosinophilia induced by interleukin (IL)-5 secreted from CD4+ T cells. However, it is still obscure whether eosinophils play an important role in the protection against N. brasiliensis infection. In this study we attempted to determine whether the in vivo environment of IL-5 transgenic mice, characterized by high eosinophil production, could affect the worm burden after N. brasiliensis infection. Kinetic studies on the infection demonstrated a significantly lower worm recovery from the intestine of IL-5 transgenic mice compared to age-matched background controls. This tendency was also observed at the lung stage of the infection. Furthermore, with respect to elevation of the serum IgE concentration, the peak level was observed at 2 weeks after infection in infected background control mice with four times higher concentrations than those of uninfected mice. In contrast, the increase of IgE concentration in IL-5 transgenic mice was very limited and low. The adoptive transfer of eosinophils from IL-5 transgenic mice into background control animals resulted in the reduction of worm recovery from the lungs, suggesting that eosinophils play a key role in the protection against migrating larvae of N. brasiliensis. These results indicate that the innate high level of eosinophils due to constitutive production of IL-5 augments immunity against N. brasiliensis infection.