Fasciola hepatica: Histology of the testis in egg-producing adults of several laboratory-maintained isolates of flukes grown to maturity in cattle and sheep and in flukes from naturally infected hosts

A total of 8 calves approximately 6 months old and 22 lambs of similar age were infected with metacercariae of Fasciola hepatica of various laboratory-maintained isolates including: Cullompton (sensitive to triclabendazole) and Sligo, Oberon and Leon (reported as resistant to triclabendazole). Ten to 16 weeks after infection, flukes were harvested from these experimental animals and the histology of the testis tissue was examined in a representative sample of flukes from each population. Adult wild-type flukes were also collected from 5 chronically infected cattle and 7 chronically infected sheep identified at post-mortem inspection. The testis tissue of these flukes was compared with that of the various laboratory-maintained isolates. Whilst the testes of the wild-type, Oberon and Leon flukes displayed all the usual cell types associated with spermatogenesis in Fasciola hepatica (spermatogonia, spermatocytes, spermatids and mature sperm), the Cullompton flukes from both cattle and sheep showed arrested spermatogenesis, with no stages later than primary spermatocytes represented in the testis profiles. The presence of numerous eosinophilic apoptotic bodies and nuclear fragments suggested that meiotic division was anomalous and incomplete. In contrast to the wild-type flukes, no mature spermatozoa were present in the testes or amongst the shelled eggs in the uterus. A high proportion of the eggs collected from these flukes hatched to release normal-appearing miracidia after an appropriate incubation period, as indeed was the case with all isolates examined and the wild-type flukes. It is concluded that the eggs of Cullompton flukes are capable of development without fertilization, i.e. are parthenogenetic. The implications of this for rapid evolution of resistant clones following an anthelmintic selection event are discussed. Amongst the Sligo flukes examined, two subtypes were recognised, namely, those flukes with all stages of spermatogenesis and mature spermatozoa present in the testes (type 1), and those flukes with all stages of spermatogenesis up to spermatids present, but no maturing spermatozoa in the testes (type 2). Each sheep infected with the Sligo isolate had both type 1 (approximately 60%) and type 2 (approximately 40%) flukes present in the population. Spermatozoa were found amongst the eggs in the uterus in 64% of flukes and this did not necessarily reflect the occurrence of spermatozoa in the testis profiles of particular flukes, suggesting that cross-fertilization had occurred. The apparent disruption of meiosis in the spermatocytes of the Cullompton flukes is consistent with reports that Cullompton flukes are triploid (3n = 30), whereas the Sligo and wild-type flukes are diploid (2n = 20). In the Sligo flukes the populations are apparently genetically heterogenous, with a proportion of the flukes unable to produce fully formed spermatozoa perhaps because of a failure in spermiogenesis involving elongation of the nucleus during morphogenesis. (C) 2008 Elsevier B.V. All rights reserved.

Role of endoreduplication and apomeiosis during parthenogenetic reproduction in the model brown alga Ectocarpus

The filamentous brown alga Ectocarpus has a complex life cycle, involving alternation between independent and morphologically distinct sporophyte and gametophyte generations. In addition to this basic haploid–diploid life cycle, gametes can germinate parthenogenetically to produce parthenosporophytes. This article addresses the question of how parthenosporophytes, which are derived from a haploid progenitor cell, are able to produce meiospores in unilocular sporangia, a process that normally involves a reductive meiotic division.
We used flow cytometry, multiphoton imaging, culture studies and a bioinformatics survey of the recently sequenced Ectocarpus genome to describe its life cycle under laboratory conditions and the nuclear DNA changes which accompany key developmental transitions.
Endoreduplication occurs during the first cell cycle in about one-third of parthenosporophytes. The production of meiospores by these diploid parthenosporophytes involves a meiotic division similar to that observed in zygote-derived sporophytes. By contrast, meiospore production in parthenosporophytes that fail to endoreduplicate occurs via a nonreductive apomeiotic event.
Our results highlight Ectocarpus’s reproductive and developmental plasticity and are consistent with previous work showing that its life cycle transitions are controlled by genetic mechanisms and are independent of ploidy.

Life history variation in Dasya ocellata (Dasyaceae, Rhodophyta)

Four of the five members of the Dasyaceae found in the British Isles, Dasya corymbifera J. Agardh, Dasya hutchinsiae Harvey, Dasya punicea Meneghini ex Zanardini and Heterosiphonia plumosa (Ellis) Batters, appear to have Polysiphonia-type life histories on the basis of evidence from field collections of tetrasporophytes and gametophytes. In collections from the British Isles of the fifth species, Dasya ocellata (Grateloup) Harvey, only tetrasporophytes have ever been observed, but there are two reports of gametophytes in this species from further south in Europe. Dasya ocellata tetraspores were isolated into culture from populations in Strangford Lough, Northern Ireland, and Agadir, Morocco, where one female thallus was collected amongst tetrasporophytes. Dasya ocellata from Ireland underwent a direct tetraspore-to-tetrasporophyte life history, which was followed through two complete cycles. Karyological studies showed that meiosis does not occur during tetrasporangial development: tetrasporangia are mitotic, with c. 64 small chromosomes. Comparison with chromosome numbers in meiotic tetrasporangia of D. hutchinsiae (n = c. 32) showed that this is the diploid chromosome complement. Tetraspores from the Moroccan isolate, by contrast, gave rise to gametophytes (although only the males became fertile) and tetrasporophyte recycling did not occur. Thalli sampled from a population in southern Portugal consisted only of tetrasporophytes. Dasya ocellata, like many members of the Ceramiales, shows intraspecific life history variability; a sexual life history apparently occurs only in southern populations.

Life history of the rare red alga Tsengia bairdii (=Platoma bairdii) (Nemastomataceae, Rhodophyta) from Scotland

A single thallus of the rare red seaweed Tsengia bairdii (Farlow) K. Fan et Y.Fan( Platoma bairdii (Farlow) Kuckuck) (Nemastomataceae) was collected on a subtidal pebble on the west coast of Scotland. The terete gelatinous axes, which were only 7 mm high, were monoecious. They bore numerous cystocarps and a few spermatangia, which represent the first observation of male structures in this genus. Released carpospores grew into expanded basal discs that gave rise to erect axes bearing irregularly cruciate tetrasporangia. irregularly cruciate to zonate tetrasporangia were also formed on these basal discs. Karyological studies on dividing tetrasporocytes showed about 25 bodies, identified as paired meiotic chromosomes on the basis of their size in comparison to mitotic and meiotic chromosomes in other red algal species. These observations confirm the isomorphic life history inferred from early field collections and show that this species is monoecious. Tsengia bairdii is an extremely rare seaweed in Europe - it seems to be confined to sublittoral cobbles and has a temporally patchy distribution.

A KARYOLOGICAL DEMONSTRATION OF MEIOSIS IN GELIDIUM-LATIFOLIUM (GELIDIACEAE, RHODOPHYTA) FROM IRELAND

A population of Gelidium latifolium (Greville) Bornet et Thuret (Rhodophyta) from Portstewart, County Antrim, Northern Ireland, was dominated by tetrasporophytes. When grown in culture, excised tips from 10 non-reproductive individuals all formed tetrasporangial branches. Chromosome counts in mitotic nuclei of vegetative cells from cultured tetrasporophytic apices were 58 +/- 4 chromosomes. In nuclei of dividing tetrasporocytes there were 29 +/- 2 larger bodies that were interpreted as paired meiotic chromosomes. Field-collected tetrasporophytes from Islandmagee. County Antrim. also showed approximately 29 pairs of chromosomes during meiosis in tetrasporocytes, This is the first report of meiosis in G. latifolium and the first direct demonstration of meiosis in this commercially important genus. In germinating tetraspores, the haploid nucleus initially divided prior to or during formation of the germination tube. The two daughter nuclei then underwent synchronous mitoses to form four haploid nuclei (n = 29 +/- 2), only one of which entered the germination tube. The sporeling survival rate was low, and few plants grew to maturity. The largest of these was diploid, with 55-58 chromosomes, and formed spermatangia after 14 months in culture. Other plants, which were abnormally bushy and densely branched, failed to reproduce. Since the most vigorous individual (and possibly also the other survivors) had apparently diploidized spontaneously during development, it is possible that the lack of gametophytes in the local G. latifolium population results from poor viability of haploid sporelings.

A NOVEL LIFE-HISTORY IN AGLAOTHAMNION-DIAPHANUM-SP NOV (CERAMIACEAE, RHODOPHYTA) FROM BRITTANY AND THE BRITISH-ISLES

A diminutive species of Aglaothamnion (Ceramiaceae, Rhodophyta), A. diaphanum sp. nov., is described from Brittany (Atlantic France), the Isles of Scilly (off S.W. England) and western Ireland. Aglaothamnion diaphanum is confined to the sublittoral zone, where it grows almost exclusively on algae and sessile animals attached to hard substrata. Thalli are delicate, and branched distichously in one plane. The main axes are ecorticate but may form loose non-corticating rhizoidal filaments. The lateral branches bear a characteristic, regularly alternate distichous series of branchlets, the first of which is always adaxial. All vegetative cells are uninucleate. The majority of field-collected plants bear only bisporangia, but a few bisporangial plants also form spermatangia; some male plants and a single female specimen have been collected. The spermatangial branchlets consist of 3-5 spermatangial mother cells each bearing 2-4 spermatangia, which are constricted around a central nucleus. None of the U-shaped carpogonial branches showed any sign of fertilization, and the gametangia appear to be non-functional. The bisporangia are ovoid and contain two uninucleate spores separated by an oblique curved wall. The occurrence of bisporangia and the lack of adherent cortication distinguish A. diaphanum from two similar species, Aglaothamnion bipinnatum (P. Crouan et H. Crouan) Feldmann-Mazoyer and Aglaothamnion decompositum (J. Agardh) Halos. The life history in culture of French and Irish isolates of A. diaphanum consists of a series of bisporangial generations, a single plant of which also formed spermatangia. Apical cells of bisporophytes are haploid (n = c. 32), but the first division of meiosis, with chromosome pairing and crossing over, occurs in dividing bisporocytes. The germinating bispores are haploid. Endodiploidization may occur in the early stages of sporangium development, as in some phycomycete fungi, or in vegetative cells that subsequently give rise to bisporocytes. This is the first demonstration in the red algae of meiotic bisporangia on plants of which the apical cells, at least, are haploid.

A cell surface antigen associated with meiosis in male Staurdoderus scalaris (Orthopteran)

Immunofluorescence has identified seven monoclonal antibodies reactive with the surface of meiotic cells and absent in premeiotic cells. Analysis by immunogold electron microscopy indicated that these antigens were present on the external surface of the cells and were coincident with the presence of synaptonemal complexes in the nucleus. On immunoblots a common glycosylated protein of 205 kDa was recognized, in addition to smaller subunits, suggesting the presence of a protein complex comprised of smaller peptides.

Intracellular antigenic changes associated with meiosis in the orthopteran Stauroderus scalaris

Monoclonal antibodies have been prepared against purified pachytene cells from grasshopper testes. Immunoblotting and immunofluorescence analyses identified those monoclonal antibodies which showed specificity for antigens in pachytene cells. Several antigenic changes were found to be associated with meiotic cells. Five monoclonal antibodies detected antigens which were located in the cytoplasm of premeiotic cells but were nuclear during meiosis. One monoclonal antibody showed a discrete cytoplasmic fluorescent pattern in meiotic, but not in premeiotic, cells. Another bound specifically to the nuclei of some epithelial cells at the base of follicles in mature testes.

Chromosome synapsis and recombination in simple and complex chromosomal heterozygotes of tuco-tuco (Ctenomys talarum: Rodentia: Ctenomyidae):Rodentia: Ctenomyidae)

The chromosomal speciation hypothesis suggests that irregularities in synapsis, recombination, and segregation in heterozygotes for chromosome rearrangements may restrict gene flow between karyotypically distinct populations and promote speciation. Ctenomys talarum is a South American subterranean rodent inhabiting the coastal regions of Argentina, whose populations polymorphic for Robertsonian and tandem translocations seem to have a very restricted gene flow. To test if chromosomal differences are involved in isolation among its populations, we examined chromosome pairing, recombination, and meiotic silencing of unsynapsed chromatin in male meiosis of simple and complex translocation heterozygotes using immunolocalization of the MLH1 marking mature recombination nodules and phosphorylated histone γH2A.X marking unrepaired double-strand breaks. We observed small asynaptic areas labeled by γH2A.X in pericentromeric regions of the chromosomes involved in the trivalents and quadrivalents. We also observed a decrease of recombination frequency and a distalization of the crossover distribution in the heterozygotes and metacentric homozygotes compared to acrocentric homozygotes. We suggest that the asynapsis of the pericentromeric regions are unlikely to induce germ cell death and decrease fertility of the heterozygotes; however, suppressed recombination in pericentromeric areas of the multivalents may reduce gene flow between chromosomally different populations of the Talas tuco-tuco.

Analgesic exposure in pregnant rats affects fetal germ cell development with inter-generational reproductive consequences

Analgesics which affect prostaglandin (PG) pathways are used by most pregnant women. As germ cells (GC) undergo developmental and epigenetic changes in fetal life and are PG targets, we investigated if exposure of pregnant rats to analgesics (indomethacin or acetaminophen) affected GC development and reproductive function in resulting offspring (F1) or in the F2 generation. Exposure to either analgesic reduced F1 fetal GC number in both sexes and altered the tempo of fetal GC development sex-dependently, with delayed meiotic entry in oogonia but accelerated GC differentiation in males. These effects persisted in adult F1 females as reduced ovarian and litter size, whereas F1 males recovered normal GC numbers and fertility by adulthood. F2 offspring deriving from an analgesic-exposed F1 parent also exhibited sex-specific changes. F2 males exhibited normal reproductive development whereas F2 females had smaller ovaries and reduced follicle numbers during puberty/adulthood; as similar changes were found for F2 offspring of analgesic-exposed F1 fathers or mothers, we interpret this as potentially indicating an analgesic-induced change to GC in F1. Assuming our results are translatable to humans, they raise concerns that analgesic use in pregnancy could potentially affect fertility of resulting daughters and grand-daughters.