Did geckos ride the Palawan raft to the Philippines?

Aim We examine the genetic diversity within the lizard genus Gekko in the Philippine islands to understand the role of geography and geological history in shaping species diversity in this group. We test multiple biogeographical hypotheses of species relationships, including the recently proposed Palawan Ark Hypothesis. Location Southeast Asia and the Philippines. Methods Samples of all island endemic and widespread Philippine Gekko species were collected and sequenced for one mitochondrial gene (NADH dehydrogenase subunit 2) and one nuclear gene (phosducin). We used maximum likelihood and Bayesian phylogenetic methods to derive the phylogeny. Divergence time analyses were used to estimate the time tree of Philippine Gekko in order to test biogeographical predictions of species relationships. The phylogenetic trees from the posterior distribution of the Bayesian analyses were used for testing biogeographical hypotheses. Haplotype networks were created for the widespread species Gekko mindorensis to explore genetic variation within recently divergent clades. Results Both maximum likelihood and Bayesian phylogenetic analyses indicated that Philippine Gekko species are a diverse clade with a long history in the archipelago. Ancestral range reconstruction and divergence time analyses suggest a Palawan microcontinental origin for this clade, coinciding with Palawan’s separation from Asia beginning 30 Ma, with subsequent diversification in the oceanic Philippine islands. The widespread species G. mindorensis and G. monarchus diversified in the late Miocene/early Pliocene and are potentially complexes of numerous undescribed species. Main conclusions The view of the Philippine islands as a ‘fringing archipelago’ does not explain the pattern of species diversity in the genus Gekko. Philippine Gekko species have diversified within the archipelago over millions of years of isolation, forming a large diverse group of endemic species. Furthermore, the Philippine radiation of gekkonid lizards demonstrates biogeographical patterns most consistent with stochastic colonization followed by in situ diversification. Our results reveal the need to consider deeper time geological processes and their potential role in the evolution of some Philippine terrestrial organisms.     

Loop‐mediated isothermal amplification (LAMP) for the identification of invasive Aedes mosquito species

Invasive Aedes mosquito species (Diptera: Culicidae) are of public health concern in Europe because they are either recognized or potential vectors of pathogens. Loop‐mediated isothermal amplification (LAMP) is a rapid and simple method for amplifying DNA with high specificity and efficiency, with the technique having potential for application in the field, including in high‐throughput format. Specific LAMP assays based on rDNA internal transcribed spacers 1 or 2 sequences, considering intraspecies variability at these loci, were developed for Aedes aegypti, Aedes albopictus, Aedes japonicus, Aedes koreicus and the indigenous Aedes geniculatus. No such assays could be developed for Aedes atropalpus and Aedes triseriatus because both loci were too short to serve as target. The assays rely on the clearly visible colour change from violet to sky blue after successful amplification. Sensitivity of egg detection was confirmed with ratios of up to one mosquito egg in 99 other eggs. Simple sample preparation of adults or eggs by mechanical homogenization in water required an additional heat treatment or centrifugation step to avoid non‐specific colour changes. Thus, further technical improvements are needed to render these assays truly field‐applicable, which would greatly facilitate surveillance of these invasive mosquito species and allow for prompt implementation of control measures.     

日本囊对虾Kunitz型蛋白酶抑制剂在毕赤酵母中的表达纯化及活性分析

SKPI(shrimp Kunitz-type protease inhibitor)是日本囊对虾(Marsupenaeus japonicus)体内的一个小分子多肽, 含有一个Kunitz型结构域, 属于丝氨酸蛋白酶抑制剂。目前已知丝氨酸蛋白酶抑制剂在节肢动物免疫系统中起着非常重要的作用, 为了了解SKPI在对虾天然免疫系统中的作用, 首先对其进行了重组表达。从日本囊对虾肝胰腺中扩增skpi的cDNA片段, 插入改造后的pPIC9K酵母表达载体, 获得的重组质粒转化至毕赤酵母GS115进行表达。由于改造的pPIC9K载体加入了6-His标签, 因此利用Ni?Sepharose?High?Performance对SKPI进行了高效纯化。初步的活性研究表明, 重组表达的SKPI能特异性地抑制胰蛋白酶的水解活性。     

Effect of increased pCO2 in seawater on survival rate of different developmental stages of the harpacticoid copepod Tigriopus japonicus

The rapid increase in carbon dioxide levels in seawater is causing ocean acidification and is expected to have significant effects on marine life. To explore the ability of the harpacticoid copepod Tigriopus japonicus to adapt to an increased concentration of dissolved carbon dioxide (CO₂) in seawater, we compared the survival rates of adult and nauplius stages at 400, 1000, and 1550?ppm pCO₂ over a 14-day period. The survival rate of T. japonicus dramatically decreased over time with increase in pCO₂ concentration. At 1550?ppm, the survival rate showed a decrease of more than 20% at the end of the experimental period over that at 400?ppm. Furthermore, the survival rate decreased by a greater amount at all concentrations in nauplii than in adults, with a greater effect in wild-collected specimens than in culture-derived individuals. The results suggest that future ocean acidification may negatively influence the sustainability of T. japonicus and thus may eventually influence benthic ecosystems.     

The roles of history: age and prior exploitation in aquatic container habitats have immediate and carry‐over effects on mosquito life history

1. Per‐capita resource availability in aquatic habitats is influenced directly by consumer density via resource competition and indirectly via delayed resource competition when temporally non‐overlapping cohorts of larvae exploit the same resources. In detritus‐based systems, resources are likely to be influenced by the age of the aquatic habitat, as detritus changes in quality over time and may be replenished by new inputs. 2. For aquatic insects that exploit detritus‐based habitats, feeding conditions experienced during immature stages can influence fitness directly via effects on development and survivorship, but also indirectly by influencing adult traits such as fecundity and longevity. 3. Larval habitat age and prior resource exploitation were manipulated in a field experiment using the container mosquito Aedes triseriatus. 4. It was found that A. triseriatus from older habitats had greater larval survival, faster development and greater adult longevity. Exploitation of larval habitats by a prior cohort of larvae had a significant negative effect on subsequent cohorts of larvae by delaying development. 5. It is suggested that extended conditioning of detritus probably resulted in conversion of recalcitrant resources to more available forms which improved the quality of the habitat. 6. In a parallel study, evidence was found of carry‐over effects of habitat age and prior exploitation on adult longevity for A. triseriatus and Aedes japonicus collected from unmanipulated aquatic habitats. 7. These results indicate the importance of detritus dynamics and the discontinuous nature of resource competition in these mosquito‐dominated aquatic systems.     

Purification of Gekko Small Peptide Fraction and Its Effect of Inducing Apoptosis of EC 9706 Esophageal Cancer Cells by Inhibiting PI3K/Akt/GLUT1 Signaling Pathway

This study aimed to isolate and purify a cytotoxic extraction from Gekko japonicus, identify its components and determine its cytotoxic activity in vitro. We isolated and identified the most potent cytotoxic Gekko small peptide LH-20-15. The identification and analysis of peptide sequences of LH-20-15 were performed by de novo peptide sequencing, and two new peptides were found. LH-20-15 significantly inhibited the proliferation of human esophageal squamous carcinoma EC 9706 cells in a dose-dependent manner. Furthermore, LH-20-15 induced apoptosis in esophageal cancer cells by activating the mitochondrial apoptotic pathway. Further research showed that LH-20-15 inhibited the PI3 K/Akt/GLUT1 signaling pathway. In conclusion, LH-20-15 from Gekko japonicus is a peptide mixture and may inhibit EC 9706 cell proliferation and induce apoptosis by activating the mitochondrial apoptotic pathway. It also regulates glucose metabolism by targeting the PI3 K/Akt/GLUT1 signaling pathway. These small peptides could be new sources of natural cytotoxic ingredients against esophageal cancer with potential drug values.     

What can metazoan parasites reveal about the taxonomy of Scomber japonicus Houttuyn in the coast of South America and Madeira Islands?

The metazoan parasites of four populations of the chub mackerel Scomber japonicus were analysed from two localities in the Atlantic Ocean (Madeira Islands, Portugal, and Rio de Janeiro, Brazil) and two localities in the Pacific Ocean (Callao, Peru, and Antofagasta, Chile), collected during 2002 and 2003. A total of 373 fish specimens were studied and 34 metazoan parasite species were obtained. Parasites identified from the populations of chub mackerel studied could be separated into three categories: parasites with a wide distribution, present in the Pacific and Atlantic, parasites proper of the Pacific Ocean and parasites proper of the Atlantic Ocean. The analyses of some highly specific parasites of the genus Scomber ( i.e. monogeneans of the genus Kuhnia and didymozoid digeneans) strongly suggest the need for a revision of the taxonomic status of chub mackerels from the Atlantic and Pacific coast of America. The results demonstrated the usefulness of parasites as adequate tools to clarify the taxonomic status of their hosts.     

花鲈对不同饲料原料的表观消化率

试验以Cr2 O3 为指示物 ,以 70 %基础饲料和 30 %的待测饲料原料组成试验饲料 ,测定花鲈对红鱼粉、豆粕、花生粕、菜籽粕、肉骨粉和棉籽粕中干物质、粗蛋白、粗脂肪、能量和氨基酸的表观消化率。在水温 2 5± 1℃的试验条件下 ,花鲈对鱼粉、豆粕、花生粕的干物质表观消化率均在 6 0 %以上 ;对鱼粉、豆粕、花生粕和菜籽粕的蛋白质、脂肪的表观消化率都在 80 %以上 ;花鲈对能量的表观消化率变化范围在 16 99%— 83 96 %。测得每种原料的 16种氨基酸的表观消化率 :花鲈对原料氨基酸消化率的变化与对蛋白质消化率的变化相一致 ,除了赖氨酸、缬氨酸、蛋氨酸之外 ,花鲈对肉骨粉中氨基酸的利用率最低。花鲈对饼粕类原料中的含硫氨基酸 (如蛋氨酸 )的表观消化率低于其他氨基酸 ,对鱼粉中氨基酸利用率最低的为组氨酸 ,对肉骨粉中氨基酸利用率最低的为苏氨酸。     

蛤蚧前背侧室嵴(ADVR)的分区及细胞形态学研究

采用解剖学方法和绀织学方法仔细观察,分析蛤蚧（Gekko gecko）前背侧室嵴（ADVR）的形态学分区,发现以ADVR表面的浅沟为标记,参照细胞着色深浅,细胞密度分布特征以及细胞形态大小,可将ADVR分为内侧区（ma）、嘴外侧区（rla）和尾外侧区（cla）等3个部分,为ADVR的结构功能的深入研究提供了形态学依据。     

Possible Mechanism of Oxygen Activation in Linoleate Peroxidation by Fusarium Lipoxygenase

The Oxygen activating mechanism of Fusarium lipoxygenase, a heme-containing dioxygenase, was studied. The enzyme did not require any cofactors, such as H₂O₂, however, both superoxide dismutase and catalase inhibited linoleate peroxidation by Fusarium lipoxygenase. A low concentration of H₂O₂ caused a distinct acceleration in enzymatic peroxidation. These results indicate that both O₂? and H₂O₂ are produced as essential intermediates of oxygen activation during formation of linoleate hydroperoxides by Fusarium lipoxygenase. This peroxidation reaction was also prevented by scavengers of singlet oxygen (¹O₂), but not by scavengers of hydroxy 1 radical (OH). Generation of O₂? in the enzyme reaction was detected by its ability to oxidize epinephrine to adrenochrome. Moreover, the rate of peroxide formation was greater in the D₂O than in the H₂O buffer system. These results suggest that the Haber–Weiss reaction (O₂^?+H₂O₂→OH^?+OH^·+¹O₂) is taking part in linoleate peroxidation by Fusarium lipoxygenase, and the ¹O₂ evolved could be responsible for the peroxidation of linoleate. H₂O₂ produced endogenously in the enzyme reaction might act as an activating factor for the enzyme. This possible mechanism of oxygen activation can explain the absence of a need for exogenous cofactors with Fusarium lipoxygenase in contrast to an other heme-containing dioxygenase, tryptophan pyrrolase, which requires an exogenous activating factor, such as H₂O₂.