Isolation and characterization of seven polymorphic microsatellite loci in the box jellyfish (Chironex fleckeri,Cubozoa, Cnidaria)

Population structure and migration patterns for the highly venomous cubozoan box jellyfish (Chironex fleckeri) are unknown. We describe the development of seven polymorphic microsatellites for this species, the first such loci to be characterized for any cubozoan or indeed cnidarian species other than some corals. These markers will be suitable for use in population and behavioural genetic studies.     

Meteorona kishinouyei,a new family,genus and species (Cnidaria,Cubozoa, Chirodropida) from Japanese Waters

A new family, genus and species of cubozoan box jellyfish belonging to the order Chirodropida is reported from the eastern Japan. Meteorona kishinouyei gen. et sp. n. possesses the following unique morphological characters with respect to other known species in the Chirodropida: having one tentacle per scalpel-like unbranched pedalium and slightly raised unbranched gastric saccules. A comparative table of the primary diagnostic characters of genus and order in the Chirodropida is given. The order Chirodropida is redefined. The family Chiropsellidae is established. Discussion is provided on the implications for these findings on our current understanding of Cubozoan systematics.     

Monoclonal antibodies neutralizing the haemolytic activity of box jellyfish (Chironex fleckeri) tentacle extracts

• 1.1. Three monoclonal antibodies have been produced which neutralize in vitro the haemolytic activity present in tentacle extracts of the box jellyfish (Chironex fleckeri).
• 2.2. Two of these monoclonal antibodies bound specifically to a component of relative molecular mass 50,000 in tentacle extract on Western blots.
• 3.3. This binding only occurred when the extracts were electrophoresed under non-reducing conditions.
• 4.4. The third monoclonal antibody did not display binding to Western blots of tentacle extract under any of our experimental conditions.

     

Gonadal cnidocytes in the cubozoan Tripedalia cystophora Conant, 1897 (Cnidaria: Cubozoa)

Cubozoans have a complex lifecycle in many ways similar to the scyphozoan lifecycle. The sexual reproduction within cubozoans varies between species with one clade having copulation and internal fertilization and the release of planula larvae. This cubozoan clade, the family Tripedaliidae, includes three species, Copula sivickisi, Tripedalia cystophora, and Tripedalia binata. In a recent study, it was suggested that in C. sivickisi cnidocytes play a new and important role during the sexual reproduction. Male derived cnidocytes anchor sperm packages to the female gonads and female derived cnidocytes protect the externalized embryo strand. Here, we have examined the gonads and gametes of T. cystophora and our results reveal that the male produced spermatozeugmata have a high number of isorhiza type cnidocytes, which are transferred along with the sperm during copulation. This adds further support to our hypothesis that they are important for sperm anchorage. The female gonads are lacking cnidocytes all together showing that cnidocyte production is not just a default state of the epithelium in these animals.     

Ultrastructure of a novel eurytele nematocyst of Carybdea alata Reynaud (Cubozoa,Cnidaria)

The ultrastructural characteristics of nematocysts from the cubozoan Carybdea alata Reynaud, 1830 (Hawaiian box jellyfish) were examined using light, scanning and transmission electron microscopy. We reclassified the predominant nematocyst in C. alata tentacles as a heterotrichous microbasic eurytele, based on spine, tubule and capsule measurements. These nematocysts exhibited a prominent and singular stylet, herein referred to as the lancet. Discharged nematocysts from fixed tentacle preparations displayed the following structures: a smooth shaft base, lamellae, a hemicircumferential fissure demarking the proximal end of a stratified lancet, and a gradually tapering tubule densely covered with large triangularly shaped spines. The lancet remained partially adjoined to the shaft base in a hinge-like fashion in rapidly fixed, whole-tentacle preparations. In contrast, this structure was not observed in discharged nematocyst preparations which involved multiple transfer steps prior to fixation. Various approaches were designed to detect this structure in the absence of fixative. Detached lancets were located in proximity to discharged tubules in undisturbed coverslip preparations of fresh tentacles. In addition, examination of embedded nematocysts from fresh tentacles laid on polyacrylamide gels revealed still-attached lancets. To examine the function of this structure in prey capture, Artemia sp. laden tentacles were prepared for scanning electron microscopy. While carapace exteriors exhibited structures proximal to the lancet, i.e., the nematocyst capsule and shaft base, neither tubule nor lancet structures were visible. Taken together, the morphological data suggested a series of events involved in the discharge of a novel eurytele from C. alata.     

Eight microsatellite loci for the Irukandji syndrome-causing carybdeid jellyfish, Carukia barnesi (Cubozoa, Cnidaria)

Microsatellites are high-resolution genetic markers that may be applied to examine parentage, population structure and the direction and extent of dispersal. Here we present eight polymorphic microsatellite loci developed for the carybdeid jellyfish, Carukia barnesi. The loci were developed from a microsatellite-enriched, partial genomic DNA library and tested for polymorphism on animals from each of two geographically distinct populations, Lizard Island and Double Island, from the Great Barrier Reef. The number of alleles observed for each locus ranged from 7 to 19.     

Toxinological and immunological studies of capillary electrophoresis fractionated Chrysaora quinquecirrha (Desor) fishing tentacle and Chironex fleckeri Southcott nematocyst venoms

Repeated runs of capillary electrophoresis (CE) were used to study partially-purified jellyfish nematocyst venom protein in concentrations sufficient to perform toxinological assays. Nematocyst venoms from Chironex fleckeri (Cf) and Chysaora quinquecirrha were processed. The CE eluate was divided into quadrants by scanning protein content. The fourth fraction of both jellyfish venoms, contained proteins with the smallest molecular weight components, which were responsible for the highest hemolysins and the humoral and cell-mediated immunological activity. Cytotoxic Cf lethal factor activity against human liver cells was widely dispersed throughout both venoms but more prominent in fraction 4. A V(beta) receptor human T-cell repertoire was not species-specific for either crude or fractionated jellyfish nematocyst venom.     

Chironex fleckeri (Box Jellyfish) Venom Proteins: EXPANSION OF A CNIDARIAN TOXIN FAMILY THAT ELICITS VARIABLE CYTOLYTIC AND CARDIOVASCULAR EFFECTS*

The box jellyfish Chironex fleckeri produces extremely potent and rapid-acting venom that is harmful to humans and lethal to prey. Here, we describe the characterization of two C. fleckeri venom proteins, CfTX-A (∼40 kDa) and CfTX-B (∼42 kDa), which were isolated from C. fleckeri venom using size exclusion chromatography and cation exchange chromatography. Full-length cDNA sequences encoding CfTX-A and -B and a third putative toxin, CfTX-Bt, were subsequently retrieved from a C. fleckeri tentacle cDNA library. Bioinformatic analyses revealed that the new toxins belong to a small family of potent cnidarian pore-forming toxins that includes two other C. fleckeri toxins, CfTX-1 and CfTX-2. Phylogenetic inferences from amino acid sequences of the toxin family grouped CfTX-A, -B, and -Bt in a separate clade from CfTX-1 and -2, suggesting that the C. fleckeri toxins have diversified structurally and functionally during evolution. Comparative bioactivity assays revealed that CfTX-1/2 (25 μg kg⁻¹) caused profound effects on the cardiovascular system of anesthetized rats, whereas CfTX-A/B elicited only minor effects at the same dose. Conversely, the hemolytic activity of CfTX-A/B (HU₅₀ = 5 ng ml⁻¹) was at least 30 times greater than that of CfTX-1/2. Structural homology between the cubozoan toxins and insecticidal three-domain Cry toxins (δ-endotoxins) suggests that the toxins have a similar pore-forming mechanism of action involving α-helices of the N-terminal domain, whereas structural diversification among toxin members may modulate target specificity. Expansion of the cnidarian toxin family therefore provides new insights into the evolutionary diversification of box jellyfish toxins from a structural and functional perspective.     

Growth, development and temporal variation in the onset of six Chironex fleckeri medusae seasons: a contribution to understanding jellyfish ecology

Despite the worldwide distribution, toxicity and commercial, industrial and medical impacts jellyfish present, many aspects of their ecology remain poorly understood. Quantified here are important ecological parameters of Chironex fleckeri medusae, contributing not only to the understanding of an understudied taxon, the cubozoa, but also to the broader understanding of jellyfish ecology. C. fleckeri medusae were collected across seven seasons (1999, 2000, 2003, 2005-07 and 2010), with growth rates, temporal variation in the medusae season onset and differences in population structure between estuarine and coastal habitats quantified. With a mean of 2 September ± 2 d (mean ± 95% confidence limits), the earliest date of metamorphosis was temporally constrained between seasons, varying by only 7 d (30 August to 5 September). Juvenile medusae appeared to be added over an extended period, suggesting polyp metamorphosis was an ongoing process once it commenced. At a maximum of 3 ± 0.2 mm d(-1) IPD, medusae growth to an asymptotic size of ~190 mm IPD was rapid, yet, with the oldest medusae estimated to be ~78 d in age, medusae did not appear to accumulate along the coastline. Furthermore, a greater proportion of juveniles were observed along the coastline, with estuarine populations typified by larger medusae. With key aspects of C. fleckeri's ecology now quantified, medusae season management protocols can be further developed.     

Evolution of the claustrum in Cnidaria: comparative anatomy reveals that it is exclusive to some species of Staurozoa and absent in Cubozoa

The claustrum in Cnidaria is a tissue in the gastrovascular cavity delimited by a central layer of mesoglea surrounded by gastrodermis (i.e., gastrodermis-mesoglea-gastrodermis), without communication with epidermis. By dividing the gastrovascular cavity, the four claustra provide an additional level of complexity. The presence of claustra in Cubozoa and Staurozoa has been used as evidence supporting a close relationship between these two cnidarian classes. However, the detailed anatomy of the claustrum has never been comparatively analyzed, rendering the evolution of this character among Cnidaria and its homology in Staurozoa and Cubozoa uncertain. This study provides a comparative investigation of the internal anatomy of the claustrum in Staurozoa and Cubozoa, addressing its evolutionary history based on recent phylogenetic hypotheses for Cnidaria. We conclude that the claustrum is a character exclusive to some species of Staurozoa, with a homoplastic evolution in the class, and that the structure called the “claustrum” in Cubozoa corresponds to the valve of gastric ostium, a structure at the base of the manubrium, which is also present in Staurozoa with and without claustrum. Thus, the claustrum cannot be a synapomorphy of a hypothetical clade uniting Staurozoa and Cubozoa, nor can its hypothetical presence in enigmatic fossils be used to support cubozoan affinities.     

玉米螟赤眼蜂对不同颜色的偏好性研究

颜色是影响昆虫行为活动的一种重要视觉信号。本研究采用方差分析和t检验测定了玉米螟赤眼蜂雌性、雄性及雌雄个体间对白、黄、橙、粉、红、蓝、绿7种颜色卡纸的选择趋性差异,结果显示,不同颜色卡纸对玉米螟赤眼蜂雌雄个体的选择趋性均存在显著性影响(F♀=76.43,P♀0.05;F♂=25.23,P♂0.05)。且以白色卡纸为对照时,玉米螟赤眼蜂雌雄个体均表现为在黄色卡纸上的选择蜂量最高,分别为对照的1.77倍和3.20倍;在同一颜色卡纸上,玉米螟赤眼蜂雌雄个体间的选择差异也不尽相同。采用相关性分析探讨玉米螟赤眼蜂选择趋性同卡纸色彩参数的相互关系,结果显示,玉米螟赤眼蜂雌雄个体选择趋性与卡纸色泽亮度及黄蓝色程度(b)均呈显著的正相关(P0.05)。研究表明,玉米螟赤眼蜂雌雄个体均偏好黄色,且不同性别个体对同一颜色的偏好程度也不尽相同。此外,物体颜色的亮度和色调均能不同程度影响玉米螟赤眼蜂的偏好性。     

瓜实蝇对虚拟波长下不同颜色的趋性（英文）

利用害虫对不同颜色的趋性进行害虫防治, 如利用黄板对实蝇的监测和防治已有很长的历史, 然而尚未见把颜色量化进行实蝇对颜色偏嗜性研究的报道。为探明对瓜实蝇Bactrocera cucurbitae最具吸引力的颜色及其虚拟波长, 本试验应用Dan Bruton的虚拟波长与RGB值的函数关系, 把RGB值转换为虚拟波长; 选择RGB值［(0, 213, 255), (0, 255, 146), (54, 255, 0), (129, 255, 0), (195, 255, 0), (255, 255, 0), (255, 190, 0)和 (255, 119, 0)］的颜色进行打印, 这些颜色对应的虚拟波长分别是480, 500, 520, 540, 560, 580, 560和600 nm; 在八面体内进行瓜实蝇对8种颜色的偏嗜性试验。结果表明： 波长在520~560 nm 之间对应的颜色对瓜实蝇的吸引率高于其他虚拟波长对应的颜色, 而540 nm (黄绿色, RGB值为 129, 255, 0)对应的颜色纸对瓜实蝇的吸引率最大。此外田间颜色偏嗜性试验也证实了黄绿色对瓜实蝇有最强的引诱作用。结果说明, 黄绿色（虚拟波长540 nm）是吸引瓜实蝇的关键颜色, 黄绿色粘虫板可作为监测与防治瓜实蝇的一种有效方法。     

黑肾卷裙夜蛾形态及生物学特性初步观察

黑肾卷裙夜蛾Plecoptera oculata Moore是我国南方珍贵树种降香黄檀Dalbergia odorifera种植区的一种食叶性害虫。【目的】初步研究黑肾卷裙夜蛾的形态特征、生物学特性以及生活史等问题。【方法】通过室内观察和野外调查,数据精确测量与统计,总结与分析了黑肾卷裙夜蛾的形态、生物学特性等问题。【结果】黑肾卷裙夜蛾幼虫6龄,黑肾卷裙夜蛾一个世代发育需要(41.64±1.59)d,越冬代约为120 d,室内饲养存活率为61.37%±0.80%,其中卵发育历期(7.53±0.40)d,孵化率为(86.67%±1.43)%,幼虫发育历期为(18.42±0.41)d,存活率为61.30%±1.21%,蛹期为(8.67±0.58)d,羽化率为75.55%±1.14%,成虫寿命为(7.02±0.78)d,存活率为74.44%±1.12%,该虫在广东地区一年发生可以发生7~8代,且具有世代重叠现象。【结论】通过了解该虫的生物学特性,为切实可行的防控黑肾卷裙夜蛾提供理论依据。