Branchiopods (non-cladocerans) of the Maltese Islands (central Mediterranean)

Four branchiopod species occur on the Maltese Islands, viz. Branchipus schaefferi, B. visnyai, Cyzicus tetracerus and Triops cancriformis. All four live in freshwater pools, but it is noteworthy that, beside B. schaefferi, T. cancriformis was recorded from a mesohaline, coast-fringing habitat.     

Comparison of the hatching process of the tadpole shrimps Triops cancriformis and Lepidurus apus lubbocki (Notostraca) and its relation to their distribution in rain-pools in Israel

Rain-pool habitats are gradually disappearing in Israel as a result of agricultural and urban development. Present and past records of notostracan distribution here reveal a difference in the occurrence of Triops cancriformis and Lepidurus apus lubbocki, the former rather rare, and support the suggestion that species of Triops are more thermophilic than Lepidurus, with optimal hatching temperature 8–12°C higher. The limited distribution of T. cancriformis in Israel may be partly attributed to sub-optimal temperatures (<20°C) in early winter.All populations of T. cancriformis in Israel were monosexual female. Resting Notostraca eggs float and become exposed to light, needed for hatching, and to warm day-time temperatures at the water surface which enhance embryonic development. Exposure to higher temperatures may be particularly important for the thermophilic T. cancriformis in Israel, where deep water temperatures in early winter are often below hatching values.     

Predation in a temporary pond with special attention to the trophic role of Triops cancriformis (Crustacea: Branchiopoda: Notostraca)

Temporary ponds are described as environments with a low predation pressure. Notostracans inhabit these types of ponds, and some populations acquire a high proportion of larger individuals, whose feeding behaviour is mainly predatory. The predation nature of Triops cancriformis is not widely accepted, because its diet is still partly controversial. We analysed the diet of one population of T. cancriformis in a Mediterranean temporary pond (Espolla pond, NE Iberian peninsula) to evaluate its predation behaviour. The gut content of the individuals bigger than 10 mm is mainly composed of detritus, plant fibres and microcrustaceans (cladocerans, ostracods and copepods). The prey number increases with the body size of individual T. cancriformis as previously described. Sex-biased predation was observed for one copepod prey, but not for the other. Predation pressure in that community was monitored along six hydroperiods as the percentage of predator biomass (not only T. cancriformis) in relation to the non-predator biomass. The proportion of predator biomass is high, and this contrasts with the low predation pressure expected for a temporary system with short hydroperiods.     

Population genetics and reproductive strategies of two Notostraca (Crustacea) species from winter ponds in Israel

Fluorescent-amplified fragment length polymorphism (FAFLP) fingerprinting assay was used to compare the genetic diversity within and between tadpole shrimps (Notostraca) populations of Lepidurus apus (n=7) and Triops cancriformis (n=2) from rain pools in Israel. Each ephemeral water body has revealed a unique fingerprint pattern with an entailed genetic drift between nearby ponds. High similarity of genotypic diversity within each geographic area led to three clusters of water bodies, north, south and center of Israel. FAFLP assays on several newly hatched individuals of T. cancriformis revealed high identity amongst kin, as compared to L. apus where newly hatched from the same maternal source showed high diversity. Results indicate that T. cancriformis populations from Israel are probably parthenogenetic as indicated by clonal structures. The higher genetic variability in the L. apus populations and in laboratory-hatched specimens indicates the existence of sexual reproduction.     

Morphological Reassessment of <Emphasis Type="Italic">Triops cancriformis</Emphasis> Bosc (Crustacea: Branchiopoda: Notostraca) Population from Kashmir,India with Comments on the Distribution of the Species in India

Triops cancriformis was reported first from India in 1871 and has been subsequently shown to occur in different regions of the country. In this paper, we have morphologically re-examined the Triops cancriformis population from Kashmir from where this species was first reported. We have compared some morphological traits of our T. cancriformis population with a few closely related species. The result shows that the Kashmir population is more similar to the typical non gonochoric Triops cancriformis in morphology. From literature records, we show that T. cancriformis has a peculiar distribution in India with no record of occurrences below 23°N. We also highlight the 125 years of history of T. cancriformis records from India.     

Gene flow and Andean uplift shape the diversification of Gasteracantha cancriformis (Araneae: Araneidae) in Northern South America

The Andean uplift has played a major role in shaping the current Neotropical biodiversity. However, in arthropods other than butterflies, little is known about how this geographic barrier has impacted species historical diversification. Here, we examined the phylogeography of the widespread color polymorphic spider Gasteracantha cancriformis to evaluate the effect of the northern Andean uplift on its divergence and assess whether its diversification occurred in the presence of gene flow. We inferred phylogenetic relationships and divergence times in G. cancriformis using mitochondrial and nuclear data from 105 individuals in northern South America. Genetic diversity, divergence, and population structure were quantified. We also compared multiple demographic scenarios for this species using a model‐based approach (Phrapl ) to determine divergence with or without gene flow. At last, we evaluated the association between genetic variation and color polymorphism. Both nuclear and mitochondrial data supported two well‐differentiated clades, which correspond to populations occurring on opposite sides of the Eastern cordillera of the Colombian Andes. The final uplift of this cordillera was identified as the most likely force that shaped the diversification of G. cancriformis in northern South America, resulting in a cis‐ and trans‐Andean phylogeographic structure for the species. We also found shared genetic variation between the cis‐ and trans‐Andean clades, which is better explained by a scenario of historical divergence in the face of gene flow. This has been likely facilitated by the presence of low‐elevation passes across the Eastern Colombian cordillera. Our work constitutes the first example in which the Andean uplift coupled with gene flow influenced the evolutionary history of an arachnid lineage.     

Retention, capture and consumption of experimental prey by orb-web weaving spiders in coffee plantations of Southern Mexico

This study focuses on the predatory capacity of four sympatric species of web- building spiders that inhabit coffee plantations in Southern Mexico: Gasteracantha cancriformis, Cyclosa caroli, and the morphologically similar species pair Leucauge mariana and L. venusta which were considered as one species group. The retention capabilities of the webs of these species and the incidence of prey capture and consumption were measured using eight types of insect prey belonging to the orders Coleoptera (1 species), Hymenoptera (3), Diptera (2) Lepidoptera (1) and Homoptera (1). The different characteristics of each prey such as body weight, body size, defensive behaviour, etc., were recorded. The incidence of prey retention, capture and consumption were significantly higher in G. cancriformis than in any of the other species. The lowest rates of retention, capture and consumption were observed in C. caroli, while L. mariana/venusta were intermediate in their predatory capabilities. Significant negative correlations between prey size and percent consumption were detected in L. mariana/venusta and in G. cancriformis; in both cases, large prey were less likely to be immediately consumed than small prey items. The results can be interpreted in the light of the morphological characteristics of the spiders. G. cancriformis possesses long legs and a carapace and appeared to have few difficulties to manipulate all types of prey. In contrast, C. caroli showed lesser abilities to manipulate and subdue aggressive prey items, perhaps due to the short leg length and unprotected body of this species. The consumption of prey items may be related to the predatory strategy of each spider. G. cancriformis constructs a new web every morning and prey storage was never observed. The absence of prey storage behaviour could explain why this species consumes prey soon after capture. In contrast, C. caroli constructs a permanent web and stores captured prey on a stabilimentum that may explain the very low incidence of immediate consumption of prey observed in this species.     

Characterization of dinucleotide microsatellite loci in the living fossil tadpole shrimp Triops cancriformis (Crustacea Branchiopoda Notostrace)

Five dinucleotide markers were obtained from a microsatellite genomic library of the tadpole shrimp Triops cancriformis. The number of alleles at each locus was low (maximum three alleles) but all loci were polymorphic. The presence of null alleles could not be disregarded. Heterozygosity levels were indicative of selfing in the studied populations whose real sexuality will be further tested using these markers.     

Ultrastructural observations on the oogenesis of Triops cancriformis (Crustacea,Notostraca)

Summary The first stages of the oogenesis of Triops cancriformis have been studied. At the outset the oocyte is smaller than the nurse cells. Meiosis begins with typical synaptonemal complexes. The significance of these complexes and of some other peculiar structures of germ cells, i.e., pore complexes and annuli within the nucleus, and annulate lamellae within the cytoplasm are discussed. The morphofunctional uniformity of some cytoplasmic structures (annulate lamellae, concentrically arranged ER, and yolk globules) in the oocyte as well as its nurse cells is also discussed.     

Recent evolution of alternative reproductive modes in the 'living fossil' <Emphasis Type="Italic">Triops cancriformis</Emphasis>

Background  

The Notostraca is a small but ancient crustacean order with a contrasting combination of a conservative morphology and a wide range of reproductive modes. The tadpole shrimp Triops cancriformis, includes bisexual – the putatively ancestral state -, androdioecious and hermaphrodite populations. As hermaphroditism and androdioecy confer a colonisation advantage, we expect the postglacial colonisation of northern Europe to have been effected by lineages with such reproductive modes. Therefore, N European populations should be composed of closely related lineages reflecting a recent range expansion. In contrast, glacial refugia in the south should contain bisexual populations with high haplotype diversity and more population structuring. To test these hypotheses, we analysed the geographic distribution of reproductive modes based on new and published sex ratio data. In addition, we investigated the European phylogeography of T. cancriformis by sequencing over a 1000 bp of mitochondrial DNA (mtDNA) in individuals from a large sample of populations of the three recognised subspecies.     

Male occurrence in Austrian populations of Triops cancriformis (Branchiopoda, Notostraca) and ultrastructural observations of the male gonad

Abstract. Recent reports of occasional males in Austrian populations of Triops cancriformis have promoted interest in an analysis of the ultrastructure of the male gonad. It appears as a double thick and well‐defined tubular structure running along the midgut, inside the hemocoel. It is composed of two portions: the wall and the lumen. The former is made up of germ cells that are widespread and intermixed with somatic cells. The lumen is ～200 μm wide and acts as the collecting site of spermatids and sperm. The germ cells are recognizable by their rounded appearance and by the occurrence of exceptionally long synaptonemal complexes in their nuclei. Their maturation implies a volume elongation and an aggregation in a characteristic “cyst” arrangement. The cysts migrate towards the lumen, where they break open, releasing spermatids and sperm. At the end of this process the lumen is filled with maturing gametes. The sperm have very condensed chromatin nuclei and partially reduced cytoplasm where the most striking features are the axoneme and strewn microtubules together with evident pseudopodia. Our morphological data support the full functionality of males in T. cancriformis. However, further work on gametogenesis, distribution of sexes, and genetic analysis of breeding experiments are needed to reach a fuller understanding of the role of males in middle and northern European populations of this species.     

Identification,expression, and molecular evolution of microRNAs in the “living fossil” Triops cancriformis (tadpole shrimp)

MicroRNAs have been identified and analyzed in various model species, but an investigation of miRNAs in nonmodel species is required for a more complete understanding of miRNA evolution. In this study, we investigated the miRNAs of the nonmodel species Triops cancriformis (tadpole shrimp), a “living fossil,” whose morphological form has not changed in almost 200 million years. Dramatic ontogenetic changes occur during its development. To clarify the evolution of miRNAs, we comparatively analyzed its miRNAs and the components of its RNAi machinery. We used deep sequencing to analyze small RNA libraries from the six different developmental stages of T. cancriformis (egg, first–fourth instars, and adult), and also analyzed its genomic DNA with deep sequencing. We identified 180 miRNAs (87 conserved miRNAs and 93 novel candidate miRNAs), and deduced the components of its RNAi machinery: the DICER1, AGO1–3, PIWI, and AUB proteins. A comparative miRNA analysis of T. cancriformis and Drosophila melanogaster showed inconsistencies in the expression patterns of four conserved miRNAs. This suggests that although the miRNA sequences of the two species are very similar, their roles differ across the species. An miRNA conservation analysis revealed that most of the conserved T. cancriformis miRNAs share sequence similarities with those of arthropods, although T. cancriformis is called a “living fossil.” However, we found that let-7 and DICER1 of T. cancriformis are more similar to those of the vertebrates than to those of the arthropods. These results suggest that miRNA systems of T. cancriformis have evolved in a unique fashion.     

Ultrastruktur und polysaccharidanteile der cuticula von Triops cancriformis Bosc. (Crustacea,Notostraca) während der Häutungsvorbereitung

Zusammenfassung Die Cuticula von Triops cancriformis besteht aus Endo-, Exo- und Epicuticula. Die Epicuticula ist aus vier Schichten aufgebaut, die Exocuticula aus 10 und die Endocuticula aus 60–80. Die Schichten der Endocuticula sind aus annähernd oberflächenparallel verlaufenden Mikrofibrillen, die zu Lamellulae zusammentreten, gebildet. Diese Lamellulae atehen senkrecht zur Oberfläche. Die Lamellulae der einzelnen Lagen verlaufen im rechten Winkel zu denen der Nachbarlagen. In Sinnesborsten verläuft so ein Teil der Fibrillen in Längsrichtung, der andere quer zur Längsachse.Polysaccharide finden sich in den Lamellulae, in zwei Schichten der Epicuticula, den Desmosomen und als Glykogengranula in Epidermiszellen.Die Häutung zeigt anscheinend keine Besonderheiten gegenüber anderen Arthropoden.

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Ultrastructure and polysaccharide content of the cuticle of Triops cancriformis Bosc. (Crustacea, Notostraca) during the Molting preparation

Summary The cuticle of Triops cancriformis consists of endo-, exo- and epicuticle. The epicuticle comprises 4 layers, the epocuticle 10 and the endocuticle 60–80 layers. The layers of the endocuticle consist of microfibrils. These microfibrils are almost parallel to the surface of the cuticle and merge into lamellulae. These lamellulae run vertical to the surface. The lamellulae in any one layer are at right angles to the lamellulae in the neighbouring layers. Thus, some of the fibrils in sensory setae are parallel to the longitudinal axis and others are perpendicular to it.Polysaccharides are found in the lamellulae, in two layers of the epicuticle, in the attachment regions and in the glycogen deposits in the epidermis cells.The molting process seems to be similar to the molting process of other arthropoda.

     

The formation of the nervous system during larval development in Triops cancriformis (Bosc) (crustacea,Branchiopoda): An immunohistochemical survey

We provide data of the development of thenervous system during the first five larval stages of Triops cancriformis. We use immunohistochemical labeling (against acetylated α‐tubulin, serotonin, histamine, and FMRFamide), confocal laser scanning microscopy analysis, and 3D‐reconstruction. The development of the nervous system corresponds with the general anamorphic development in T. cancriformis. In larval stage I (L I), all brain parts (proto‐, deuto‐, and tritocerebrum), the circumoral connectives, and the mandibular neuromere are already present. Also, the frontal filaments and the developing nauplius eye are already present. However, until stage L III, the nauplius eye only consists of three cups. Throughout larval development, the protocerebral network differentiates into distinct subdivisions. In the postnaupliar region, additional neuromeres and their commissures emerge in an anteroposterior gradient. The larval nervous system in L V consists of a differentiated protocerebrum including a central body, a nauplius eye comprising four cups, a circumoral nerve ring, mandibular‐ and postnaupliar neuromeres up to the seventh thoracic segment, each featuring an anterior and a posterior commissure, and two parallel connectives. The presence of a protocerebral bridge is questionable. The distribution of neurotransmitters in L I is restricted to the naupliar nervous system. Over the course of the five stages of development, neurotransmitter distribution also follows an anteroposterior gradient. Each neuromere is equipped with two ganglia innervating the locomotional appendages and possesses a specific neurotransmitter distribution pattern. We suggest a correlation between neurotransmitter expression and locomotion. J. Morphol., 2010. © 2010 Wiley‐Liss, Inc.     

Ultrastructural observations on the oogenesis of Triops cancriformis (Crustacea,Notostraca)

Summary Each ovarian follicle of Triops cancriformis is four-celled; these cells (one oocyte and three nurse cells) are interconnected by cytoplasmic bridges. In the course of differentiation, the nurse cells are early recognizable; they increase in size more than the oocyte and their nuclei contain many nucleoli. For the first time in Arthropoda, yolk globules are reported to be present in nurse cell cytoplasm; these globules arise from the smooth endoplasmic reticulum. The functional significance of the intercellular bridges and the trophic role of the nurse cells are discussed.The authors are grateful to Dr. Bruno Sabelli for his support and to Mr. Francesco Monte for his technical assistance     

pH-related Hatching Success of Triops cancriformis (Crustacea: Branchiopoda: Notostraca)

Six different pH ranges were tested for the hatching rate of Triops cancriformis cysts. Cumulative hatching numbers approach to approximately 50% of cysts. The highest hatching rates, co-occurring with the shortest hatching times, were recorded in the neutral range. No hatching took place at pH values 9 and above. This corresponds with the most frequent habitats of this species, snowmelt pools of river flood plains as well as aestival pools caused by heavy rainfalls. Occurrences in sodic alkaline pans can be explained by relatively low pH values at the time of inundation.     

Ultrastruktur und Funktion der Hautdrüsen vonTriops cancriformis Bosc. (Crustacea,Notostraca)

Zusammenfassung Die Hautdrüsen vonT. cancriformis bestehen aus drei Zellen: Einer Sekretbildungszelle, einer Halszelle und einer Gangzelle. In der Sekretbildungszelle werden von Golgiapparaten amorphe Tropfen bis 1m Durchmesser gebildet, die sich dann zu Lamellenstapeln differenzieren. Diese werden durch einen Ausführgang, der von Halszelle und Gangzelle gebildet wird und mit Epicuticula ausgekleidet ist, an die OberflÄche des Tieres abgegeben. Dort bilden bzw. erneuern sie die obersten Lagen der Epicuticula, auf der sie sich in dünner Schicht ausbreiten.

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Ultrastructure and function of dermal glands ofTriops cancriformis Bosc. (Crustacea, Notostraca)

Summary The dermal glands ofT. cancriformis are formed by three cells, the secretory cell, the intermediate cell and the duct cell. In the secretory cell amorphous droplets up to 1m diameter are produced by the Golgi complexes. These droplets are developed to form densely packed lamellae. These are transported to the surface of the animal via a duct, which is built up by the duct cell and the intermediate cell. The duct is lined by epicuticula. On the surface the lamellae spread out to form a thin film, the outer layers of the epicuticula.

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Liste der Abkürzungen ACU Alte Cuticula - NCU Neue Cuticuk - EP Epidermis - GZ Gangzelle - HZ Halszelle - SBZ Sekretbildungszelle - S Sekret Meinem verehrten Lehrer, Herrn Prof. Dr. V. Schwartz, zum 70. Geburtstag gewidmetTeil einer yon der FakultÄit für Bio- und Geowissenschaften der Universidit Karlsruhe angenommenen Habilitationsschrift (Rieder, 1974)     

Ontogenetic sequence comparison of extant and fossil tadpole shrimps: no support for the “living fossil” concept

The tadpole shrimp Triops cancriformis (Branchiopoda, Eucrustacea) is often referred to as a “living fossil.” This term implies that the morphology of a species has barely changed for hundreds of millions of years; in the case of T. cancriformis, for about 200 million years. In 1938, Trusheim documented fossil notostracans from the Upper Triassic of southern Germany (237–200 million years) and named them T. cancriformis minor due to their small size compared to modern forms of T. cancriformis. We compared the ontogenetic sequence of the fossil forms to that of modern forms. Fossil material came from the Museum Terra Triassic in Euerdorf and originated from the same geological formation (the Hassberge Formation) as the Trusheim material, which is considered to be nearly entirely lost. The specimens were documented using cross-polarized light and processed into high-resolution images. Fluorescence microscopy was used to document exuviae and carcasses of extant representatives of T. cancriformis. Both forms showed an elongation and similar trends in the length/width ratio of the shield during ontogeny. However, differences were found in the starting point of the developmental processes. Fossil forms start out with a more roundly shaped shield, which becomes more elliptical, while extant forms already start with a more elliptical shield shape. Further differences between extant and fossil forms were found upon comparing shield to trunk ratios. All differences are highly significant statistically. These differences in ontogeny cast severe doubt on the interpretation that T. cancriformis has been static for 237 million years. While the term “living fossil” is misleading and its use should be discouraged in general, it seems to be especially inappropriate to apply it to T. cancriformis.     

Molecular characterization of the tadpole shrimp Triops (Branchiopoda: Notostraca) from the Baja California Peninsula,México: New insights on species diversity and phylogeny of the genus

Using sequence analyses of fragments of the small and large subunits of mitochondrial genes 12S and 16S rRNA, we studied the molecular identity of five Triops populations from the Baja California Peninsula, México. Additionally, we explored the phylogeny of the genus by comparing with sequence data from gonochoric T. longicaudatus (Zacatecas, México), commercial Triops kit (U.S.A.), T. `granarius' (Japan), T. cancriformis (Austria), T. australiensis (Australia) and Lepidurus lemmoni (U.S.A.). The 16S fragment was not useful to discriminate the American Triops forms because their sequences were more than 99% similar. Molecular and phylogenetic analyses using the 12S gene fragments, in agreement with previous allozyme studies, indicate that the nominal (morphological) species T. longicaudatus is a mixture of several species such that, of the seven Triops American populations studied, six phylogenetic species can be identified and two morphologically and reproductively highly divergent forms can be grouped into a single monophyletic clade. The molecular data, rather than supporting our previous proposal that the phylogenetic relationships of Triops species could be deduced by similarities in the number of total and legless rings, suggest that T. cancriformis may represent an independent group from the rest of the species in that genus. In spite of detectable differences among American populations, our analyses indicate these represent a single monophyletic group when compared to Triops from outside of the New World.     

R2 dynamics in Triops cancriformis (Bosc, 1801) (Crustacea,Branchiopoda, Notostraca): turnover rate and 28S concerted evolution

The R2 retrotransposon is here characterized in bisexual populations of the European crustacean Triops cancriformis. The isolated element matches well with the general aspects of the R2 family and it is highly differentiated from that of the congeneric North American Triops longicaudatus. The analysis of 5′ truncations indicates that R2 dynamics in T. cancriformis populations show a high turnover rate as observed in Drosophila simulans. For the first time in the literature, though, individuals harboring truncation variants, but lacking the complete element, are found. Present results suggest that transposition-mediated deletion mechanisms, possibly involving genomic turnover processes acting on rDNAs, can dramatically decrease the copy number or even delete R2 from the ribosomal locus. The presence of R2 does not seem to impact on the nucleotide variation of inserted 28S rDNA with respect to the uninserted genes. On the other hand, a low level of polymorphism characterizes rDNA units because new 28S variants continuously spread across the ribosomal array. Again, the interplay between transposition-mediated deletion and molecular drive may explain this pattern.