Platyhelminth phylogenetics - a key to understanding parasitism?

The comparative method, the inference of biological processes from phylogenetic patterns, is founded on the reliability of the phylogenetic tree. In attempting to apply the comparative method to the understanding of the evolution of parasitism in the phylum Platyhelminthes, we have highlighted several points we consider to be of value along with many problems. We discuss four of these topics. Firstly, we view the group at a phylum level, in particular discussing the importance of establishing the sister taxon to the obligate parasite group, the Neodermata, for addressing such questions as the monophyly, parasitism or the endo or ectoparasitic nature of the early parasites. The variety of non-congruent phylogenetic trees presented so far, utilising either or both morphological and molecular data, gives rise to the suggestion that any evolutionary scenarios presented at this stage be treated as interesting hypotheses rather than well-supported theories. Our second point of discussion is the conflict between morphological and molecular estimates of monogenean evolution. The Monogenea presents several well-established morphological autapomorphies, such that morphology consistently estimates the group as monophyletic, whereas molecular sequence analyses indicate paraphyly, with different genes giving different topologies. We discuss the problem of reconciling gene and species trees. Thirdly, we use recent phylogenetic results on the tapeworms to interpret the evolution of strobilation, proglottization, segmentation and scolex structure. In relation to the latter, the results presented indicate that the higher cestodes are diphyletic, with one branch difossate and the other tetrafossate. Finally, we use a SSU rDNA phylogenetic tree of the Trematoda as a basis for the discussion of an aspect of the digenean life-cycle, namely the nature of the first intermediate host. Frequent episodes of host-switching, between gastropod and bivalve hosts or even into annelids, are indicated.

Chronostratigraphy of the CRP-3 drillhole, Victoria Land Basin, Antarctica

An 823 m thick glaciomarine Cenozoic section sitting unconformably on the Lower Devonian Beacon Supergroup was recovered in CRP –3. This paper reviews the chronostratigraphical constraints for the Cenozoic section. Between 3 and 480.27 mbsf 23 unconformity bounded cycles of sediment were recorded. Each unconformity is thought to represent a hiatus of uncertain duration. Four magnetozones have been recognised from the Cenozoic section. The record is complex with several “tiny wiggles” recorded throughout. Biostratigraphical or Sr ages, which could be used to link these magnetozones to the magnetic polarity time scale are restricted to the upper 190 m of sediment. Two diatom datums (Cavitatus jouseanus at 48.9 mbsf and Rhizosolenica antarctica at 68.60 mbsf), together with five Sr-isotope dates derived from molluscan fragments taken from between 10.88 and 190.29 mbsf indicate an early Oligocene (c. 31 Ma) age for this interval. The appearance of a new species of the bivalve ?Adamussium at about 325 mbsf, suggests that the Oligocene age can be extended down to this level. This confirms that the dominantly reversed magnetozone (R1), recorded down to about 340 mbsf, is Chron C12r. The ages imply high sedimentation rates and only minimal time gaps at the sequence boundaries. Below 340 mbsf there are no independent datums to guide the correlation of the magnetozones to the magnetic polarity time scale. However, the absence of in situ dinocysts attributable to Transantarctic Flora, if not a result of environmental control, limits the age of the base of the hole to between c. 33.5 and 35 Ma.

Molluscan stable isotope temperature estimates of the southwestern Ross Sea during the early oligocene and early miocene, CRP-2/2A and CRP-3, Victoria Land Basin, Antarctica

Stable isotope analyses of marine bivalve growth increment samples have been used to estimate early Oligocene (29.4 - 31.2) Ma and early Miocene (24.0 Ma) seafloor palaeotemperatures from the southwestern continental margin of the Ross Sea. Measured δ18O values average +2.5‰ in the early Miocene and range between +1.26 to +3.24‰ in the early Oligocene. The results show that palaeoceanographic conditions in McMurdo Sound during the mid-Cenozoic were significantly different from those of today. The minimum estimated spring through late summer seasonal temperature range was 3°C during the early Miocene and between 1 and 5°C during the early Oligocene. This compares to the equivalent modern day range of

Spermatozoal ultrastructure of spiny oysters (Spondylidae, Bivalvia) including a comparison with other bivalves

Sperm ultrastructure in three representative species of the marine bivalve family Spondylidae (spiny or thorny oysters) is examined and compared with available data on other bivalves, especially other families of the subclass Pteriomorphia. Spondylid spermatozoa are of the externally fertilizing aquasperm. type (ect-aquasperm). The acrosomal vesicle is conical with a deep basal invagination extending almost the full length of the vesicle. Vesicle contents are divisible into an inner, highly electron-dense anterior layer and a less dense posterior layer. The anterior layer is folded back on itself posteriorly and exhibits radiating plates (best developed peripherally). The vesicle rests on, and is partially embedded in, an extensive granular deposit of subacrosomal. material at the nuclear apex. This deposit extends partly into acrosomal vesicle invagination and also fills a broad depression in the anterior of the nucleus. No pre-formed axial rod (perforatorium) is present. The nucleus is round-pyriform and its contents coarsely fibrogranular. At the base of the nucleus, four broad depressions partially accommodate the midpiece mitochondria. The midpiece consists the four spherical mitochondria and the proximal and distal centrioles. The centrioles are arranged at approximately 90degrees to each other, and each consists of nine, angularly-oriented, microtubular triplets embedded in a granular matrix. A short, periodically banded rootlet connects the proximal centriole to the nuclear fossa, whereas the distal centriole, which forms the basal body to the flagellar axoneme, is anchored to the plasma membrane by nine terminally forked satellite fibres. Extensive deposits of putative glycogen rosettes surround the centrioles and mitochondria. The flagellum consists of a 9+2 axoneme sheathed by the plasma membrane. Spondylid spermatozoa strongly resemble those of the Pectinidae, further confirming the traditional view (based on comparative anatomy and shell morphology) of a close relationship between the Spondylidae and the Pectinidae. Differences in acrosomal shape and dimensions were noted between the three species examined, indicating potential taxonomic utility for comparative sperm ultrastructure within the Spondylidae.

Geographic variation of natural products of tropical nudibranch Asteronotus cespitosus

Extracts of the dorid nudibranch Asteronotus cespitosus from two geographically separate regions of Australia and from the Philippines were compared using thin-layer, high-performance liquid and gas chromatography and H-1 NMR analysis. Halogenated metabolites were detected in all mollusk specimens. The major component detected in digestive tissue of specimens from the Great Barrier Reef in northeastern Australia was 4,6-dibromo2-(2',4'-dibromophenoxy)phenol (1), with minor amounts of 3,5-dibromo-2(3',5'-dibromo-20-methoxyphenoxy)phenol (2). In a specimen collected from northwestern Australia, only 3,5-dibromo-2-(3',5'-dibromo-2'-methoxyphenoxy)phenol was found. The specimen from the Philippines contained 2,3,4,5-tetrabromo-6-(2'-bromophenoxy) phenol (3) together with a novel chlorinated pyrrolidone (4). In addition, the sesquiterpenes dehydroherbadysidolide (5) and spirodysin (6) were detected in the digestive organs and mantle tissue of the nudibranchs from the Great Barrier Reef and from the Philippines, whereas these chemicals were not found in the specimen from northwestern Australia. All of the chemicals (1-3,5, and 6) have previously been isolated from the sponge Dysidea herbacea, as have chlorinated metabolites related to 4. This is the first time the characteristic halogenated metabolites that typify Dysidea herbacea have been reported from a carnivorous mollusk, which implies a dietary origin as opposed to de novo synthesis.

The development of a positive non-infectious control for the detection of Perkinsus using the Ray test

To establish a noncontagious control for the Ray thioglycollate test for the detection of Perkinsus in mollusks we evaluated nonviable stages of P. olseni for enlargement of hypnospores and blue/black iodine stain. Trophozoites made nonviable with formalin, irradiation or colchicine failed to swell in thioglycollate. They remained small and did not differentially stain in iodine. Trophozoites that had already developed into hypnospores in thioglycollate were rendered inactive by freezing, ethanol or formalin immersion. They retained their iodinophilic properties and thus could provide a partial control for the Ray Test.

Sponge halogenated natural products found at parts-per-million levels in marine mammals

Several unknown, abundant brominated compounds (BCs) were recently detected in the blubber of dolphins and other marine mammals from Queensland (northeast Australia). The BC were interpreted as potential natural products due to the lack of anthropogenic sources for these compounds. This study investigated whether some of the BCs accumulated by diverse marine mammal species are identical with natural BCs previously isolated from sponges (Dysidea sp.) living in the same habitat. Isolates from sponges and mollusks (Asteronotus cespitosus) were compared with the signals detected in the mammals' tissue. Mass spectra and gas chromatography retention times on four different capillary columns of the isolates from sponges and mammals were identical in all respects. This proves that the chemical name of the compound previously labeled BC-2 is 4,6-dibromo-2-(2'-dibromo)phenoxyanisole and that the chemical name of BC-11 is 3,5-dibromo-2-(3',5'-dibromo-2'-methoxy)phenoxyanisole. Using a quantitative reference solution of BC-2, we established that the concentrations of the brominated metabolies found in the marine mammals are frequently >1 mg/kg. The highest concentration (3.8 mg/kg), found in a sample of pygmy sperm whale (Kogia breviceps), indicates that BC-2 is a bioaccumulative, natural organohalogen compound. This is supported by the concentrations of the BCs in our samples being equal to the highest concentrations of anthropogenic BCs in any environmental sample. The quantitative determination of BC-2 in blubber of marine mammals from Africa and the Antarctic suggests that BC-2 is wide-spread. These results are direct proof that marine biota can produce persistent organic chemicals that accumulate to substantial concentrations in higher trophic organisms.

Biologia e abundância do camarão Plesionika edwardsii (Brandt, 1851) nos Açores

Dissertação de Mestrado, Estudos Integrados dos Oceanos, 15 de Janeiro de 2009, Universidade dos Açores.

Contributions à la Paléontologie du Miocène moyen continental du Bassin du Tage. I - Quelques mollusques - Pero Filho, Póvoa de Santarém, Sítio do Mirante

Some land and freshwater mollusks (Gastropoda, Pelecypoda) from three middle Miocene localities: Pêro Filho, Póvoa de Santarém, Sítio do Mirante, all in Ribatejo province, Portugal, are summarily studied. On a systematical viewpoint it has been shown (see also Tableau): - the presence of the genus Janulus, whose species only survive now at Madeira and Canary's islands; - the presence of "Helix" cotteri that may indeed belong in the genus Megalotachea, common in western Europe since "Helvetian" to Messinian times; - Limax, Testacella, Acroloxus and Pseudamnicola are quoted for the first time in portuguese tertiary formations; Ferrisia has been identified for the first time in Iberian Peninsule; - the presence of other forms previously quoted by ANTUNES & ZBYSZEWSKI (1973), is confirmed: Bithynia, Theodoxus, Pisidium. Such faunules are compatible with a middle Miocene age. On a paleobiogeographical viewpoint, some forms suggest mediterranean affinities. Fossil associations do correspond ecologically to either palustrine or stagnant, calm and shallow water conditions (Sítio do Mirante, and essentially also Póvoa de Santarém), or to fluviatile conditions (Pêro Filho).

Contributions à la Paléontologie du Miocène moyen continental du Bassin du Tage. III - Mammifères - Póvoa de Santarém, Pêro Filho et Chões (Secorio). Conclusions générales

This study on middle Miocene mammalian faunas from Tagus'basin deals particularly with some small mammals hitherto undescribed from Portugal, including a new Glirid (Paraglirulus scalabicensis nov. sp.); it allows an accurate datation by biostratigraphical standards, Megacricetodon crusafonti, Fahlbuschia darocensis, Cricetodon jotae being characteristic of mammalian MN6 unit, thus their age is nearly that of Sansan and Manchones (however the presence of Peridyromys hamadryas and Lagopsis verus do suggest, amidst this biozone, a somewhat later age than Sansan's); it contributes with indirect correlation data with marine formations, as underlying oyster-bearing beds most probably are in correspondance to the apogee of the same transgression that deposed near Lisbon ”schlier" facies from VI-a division (Serravalian, Blow's zones 10-13, Globorotalia meyeri zone); the diversity of mammalian assemblages is surely related to an environement with varied biotopes, whose characterisation becomes easier if account is taken of the preceding papers on mollusks (G. Truc) and Cyprinid fishes (J. Gaudant), and also according to some unpublished paleobotanical data (J. Pais). A table with a synthesis of all paleontological data so far known is presented.

Evaluation of the potential of translocated common cockle for ecological risk assessment studies: bioaccumulation and biomarkers test

Thesis submitted to the Faculdade de Ciências e Tecnologia to obtain the Master’s degree in Environmental Engineering, profile in Ecological Engineering

High-resolution correlation of coastal cliff sections in the Lagos- Portimao Formation (Lower - Middle Miocene, central Algarve, Portugal)

This paper describes a high-resolution stratigraphic correlation scheme for the early to middle Miocene Lagos-Portimão Formation of central Algarve, southern Portugal. The Lagos Portimão-Formation of central Algarve is a 60 m thick package of horizontally bedded siliciclastics and carbonates. The bryozoan and mollusc dominated biofacies is typical of a shallow marine, warm-temperate climatic environment. We define four stratigraphic marker beds based on biofacies, lithology, and gamma-ray signatures. Marker bed 1 is a reddish shell bed composed predominantly of bivalve shells in various stages of fragmentation. Marker bed 2 is a fossiliferous sandstone / sandy rudstone characterized by bryozoan masses. Marker bed 3 is also a fossiliferous sandstone with abundant larger foraminifers and foliate bryozoans. Marker bed 4 is composed of three distinct layers; two fossiliferous sandstones with an intercalated shell bed. The upper sandstone unit displays thickets of the bryozoan Celleporaria palmate associated with the coral Culizia parasitica. This stratigraphic framework allows to correlate isolated outcrops within the stratigraphic context of the Lagos-Portimão Formation and to establish high resolution chronostratigraphic Sr-isotopic dating.

Susceptibility of Biomphalaria tenagophila and Biomphalaria glabrata from the same region to two Brazilian strains of Schistosoma mansoni

B. tenagophila snails from Ouro Branco, MG, showed positivity for S. mansoni, with infection rates of 5%, 10%, (SJ strain), and 1% (LE strain) using a pool of miracidia. The mollusks were found to be susceptive from the 3rd generation reared in laboratory onwards. The B. tenagophila (OB, MG) when individually exposed to 10 miracidia, showed infection rate of 2% for LE strain. B. glabrata snails from Gagé, MG, showed a positivity rate of 58% for S. mansoni (LE strain), under experimental conditions. The B. tenagophila from Cabo Frio, RJ and B. glabrata from Belo Horizonte, MG used as a control for SJ strain showed infection rates of 47% - 85% and 36% respectivily. For the LE strain, B. glabrata (BH, MG) used as control showed infection rate of 40% - 75%.

Gamma 60Co DL50/30 of Biomphalaria glabrata (Say, 1818)

The variation of resistance to 60Co gamma-rays of Biomphalaria glabrata was studied. A population of 480 mollusks was observed during 30 days - distributed in 8 groups of snails isolated and 8 groups of snails in colonies - after exposure (30 snails per group per dose) to increasing doses of gamma radiation. Doses of 10, 20, 40, 60, 80, 160, 320 and 640 Gy from a Gamma-cell 60Co irradiator, were applied to the test groups and two groups control (non-irradiated) of snails - isolated and colony - were kept apart. After have been exposed, the snails were drew back to the aquaria where they were maintained before. The survival was estimated on a daily score of the alive animals in each group-dose, starting after the irradiation exposure day. As a result, the survival self-fertilization forms (DL50/30 = 218.2 Gy) was found greater than in cross-fecundation forms. These data point to a low radio-resistance on the cross-fertilization forms - the sexual reproductive form - which is most found in nature. The lower radio-resistance of the cross-fertilization forms suggests the presence of some sex-linked hormonal factor related to this phenomenon.

Vibrio vulnificus as a health hazard for shrimp consumers

Over the last 30 years, a number of Vibrio species found in the aquatic environment have been indicated as cause of disease in human beings. Vibrio vulnificus is an emergent pathogen, an invasive and lethal marine bacterium related to wound infection and held accountable for gastroenteritis and primary septicemia. It occurs quite frequently in marine organisms, mainly in mollusks. This study aimed at isolating and identifying strains of V. vulnificus based upon the analysis of twenty samples of seabob shrimp, Xiphopenaeus kroyeri (Heller), purchased at the Mucuripe fish market (Fortaleza, Brazil). TCBS agar was used to isolate suspect strains. Seven of twenty-nine strains isolated from six different samples were confirmed as such by means of biochemical evidence and thus submitted to biological assays to determine their virulence. The susceptibility of the V. vulnificus strains to a number of antibiotics was tested. None of the V. vulnificus strains showed signs of virulence during a 24-hour observation period, possibly due to the shedding of the capsules by the cells. As to the results of the antimicrobial susceptibility tests, the seven above-mentioned V. vulnificus strains were found to be sensitive to nitrofurantoin (NT), ciprofloxacin (CIP), gentamicin (GN) and chloramphenicol (CO) and resistant to clindamycin (CI), penicillin (PN) and ampicillin (AP).