Identification and characterization of myeloma-associated antigens in Trichinella spiralis

To investigate the presence of myeloma-associated antigens in Trichinella spiralis and their anti-tumor effect, cross-immune responses between antigens of the myeloma cell SP2/0 versus positive sera to T. spiralis, and antigens of T. spiralis versus positive sera to myeloma cell SP2/0 were determined using T. spiralis and myeloma specific enzyme-linked immunosorbent assays (ELISA). The myeloma-associated antigens in T. spiralis were separated by ultrafiltration and 2-D electrophoresis, and the amino acid sequences and molecular weights were determined by spectrometry. An obvious reaction was found between a 33 kDa antigen and positive sera, and the major component of the antigen was tropomyosin (TM), which is an surface acidic protein with 284 amino acids. Mice were immunized with TM to determine the anti-tumor effect in vivo. The results showed that CD4⁺, CD8⁺ T lymphocyte, and CD19⁺ B lymphocyte were significantly increased (P < 0.05). The anti-tumor effects were significantly different between mice immunized with the antigens or adjuvant alone (P < 0.05), while the difference between mice immunized with antigens and whole T. spiralis was not significant (P > 0.05). The results indicated that TM identified in this study may play a role in eliciting cross-protective immunity.     

Trichinella spiralis: infection reduces airway allergic inflammation in mice

In an effort to define the mechanism underlying the host immune downregulation inherent to Trichinella spiralis infection, we compared the levels of Th1, Th2, and regulatory cytokines and CD4⁺CD25⁺ forkhead box P3 (FoxP3)⁺ T (T_reg) cell recruitment, as well as cellular pathology in the airway between T. spiralis infected and uninfected asthma-induced mice. After the induction of allergic airway inflammation, we noted influxes of inflammatory cells into the peribronchial tree. However, in the T. spiralis infection groups, cellular infiltration was minimal around the bronchial tree, with only a smattering of inflammatory cells. In the OVA-challenged group after T. spiralis infection, the numbers of macrophages and eosinophils in the bronchial alveolar lavage fluid were reduced by 23% and 52%, respectively, as compared to those of the OVA-challenged group. Airway hyperresponsiveness of OVA-challenged mice after T. spiralis infection was significantly suppressed as compared to the OVA-only challenged mice. The T. spiralis-infected mice exhibited a significant reduction in IL-5 concentrations relative to that noted in the OVA-challenged group (p < 0.01). Nevertheless, the regulatory cytokines IL-10 and TGF-β levels were increased significantly as the result of T. spiralis infection, and we verified the recruitment of T_reg cells in lung draining lymph nodes via T. spiralis infection. Therefore, T_reg cells, which were recruited by T. spiralis infection, might ameliorate lung function and reduce allergic airway inflammation.     

Effects of light heterogeneity on growth of a submerged clonal macrophyte

Many studies have examined the effects of spatial heterogeneity in light supply on clonal plants in terrestrial environments, but few have examined those in aquatic conditions. In a greenhouse experiment, we grew the rhizomatous submerged macrophyte Vallisneria spiralis L. in containers in three homogeneous light treatments (100%, 65%, and 30% of ambient light coded as high, medium, and low light, respectively) and two heterogeneous ones differing in patch scale (small and large patch). The growth of V. spiralis decreased significantly with decreasing light availability. In the low light conditions, V. spiralis allocated greater biomass to shoots and developed elongated leaves. In the patchy treatments, ramets distributed in the light‐rich patches had significant costs in the large patch treatment, but not in the small patch treatment, while both small and large patch treatments had no significant benefits in the light‐poor patches. We conclude that V. spiralis could escape from adverse habitats and occupy the favorable habitats. Providing the same amount of light, responses of V. spiralis to different patch scales were different at the patch level, but not at the whole clone level. Together, growth of V. spiralis could not benefit from different patch scales in heterogeneous environments at the patch as well as the whole clone level.     

Physiological integration helps a clonal macrophyte spread into competitive environments and coexist with other species

Physiological integration may help clonal macrophytes invade or escape from existing communities. No studies have tested the above hypothesis in aquatic plants. In an outdoor pond experiment, we subjected clonal fragments of the submerged macrophyte Vallisneria spiralis L. to heterogeneous environments in which V. spiralis spread from bare habitats towards vegetated habitats occupied by Myriophyllum spicatum L. or V. spiralis spread from vegetated habitats towards bare habitats. V. spiralis stolons between ramets in bare habitats and in vegetated habitats were either intact or severed. We investigated the habitat selection of V. spiralis by examining the allocation of biomass and ramets to heterogeneous habitats during its vegetative spread phase. Results showed that the stolon connection had different effects on the habitat selection of V. spiralis with regard to invasion and escape. When V. spiralis spread from bare to vegetated habitats, in comparison to severing the stolon, the stolon connection eventually facilitated a 49% increase in biomass and a 27% increase in number of ramets allocated to vegetated habitats. However, when V. spiralis spread from vegetated to bare habitats, biomass and ramets allocated to bare habitats were not significantly changed by the stolon connection (only a 5% increase in biomass and a 6% increase in number of ramets). These results indicate that clonal integration facilitated V. spiralis not to escape from but invade into vegetated habitats. The study provides evidence that physiological integration is important for survival and tolerance of ramets in competitively stressful environments and can help clonal macrophytes coexist with other species.     

Water bird guilds and their feeding connections in the Bodrogzug, Hungary

Thalli of the intertidal Phaeophyte Fucus spiralis L. and the subtidal Chlorophyte Ulva olivascens Dangeard were exposed to artificial UV-A, UV-B and photosynthetically active radiation (PAR) by combination of PAR + UV-A + UV-B (PAB), PAR + UV-A (PA) and PAR (P) treatments. UV-A enhanced photosynthesis and stimulated carbonic anhydrase (CA) and nitrate reductase (NR) in F. spiralis whilst PAR only had an inhibitory effect in this species. U. olivascens suffered chronic photoinhibition in all the treatments as evidenced by reduced maxima photosynthesis (P_max) and photosynthetic efficiency (α). Non stimulatory effect was observed upon CA and NR in this species. Our results showed that artificial UV radiation triggered opposite responses in both species. We suggest that differences shown by both species might be related to their location in the rocky shore and their ability to sense UV. We propose that the ratio UV:PAR acts as an environmental signal involved in the control of photosynthesis as shown by pronounced inhibition in samples exposed to only PAR. We also suggest that UV-regulated photosynthesis would be related to carbon (C) and nitrogen (N) cycles, regulating feedback processes that control C and N assimilation.     

Competitive ranks of three<Emphasis Type="Italic"> Fucus</Emphasis> spp. (Phaeophyta) in laboratory experiments—testing of Keddy's competitive hierarchy model

Keddy's competitive hierarchy model describes species distribution patterns along gradients under equilibrium conditions and can potentially serve as an explanation for zonation patterns of intertidal seaweeds on rocky shores. One of the assumptions of the model is a competitive hierarchy with the top competitor occupying the benign end of the gradient. Another assumption is the consistency of competitive ranks of species in all environmental conditions included in the shared parts of species' fundamental niches. In laboratory replacement series experiments, the competitive ranks of pairs of Fucus species that occupy adjacent zones in the field were analysed and compared to ranks found in previous field experiments. Unattached thalli of Fucus serratus versus F. vesiculosus or F. vesiculosus versus F. spiralis, respectively, were held in aerated beakers to establish the competitive ranking of the three congeners. Each replacement series was conducted at three total densities. F. vesiculosus was clearly competitively dominant over F. serratus. In competition with F. spiralis, F. vesiculosus was only dominant at its lowest absolute input frequencies, but at higher frequencies dominance was reversed. At high densities, the total ranking was F. spiralis > F. vesiculosus > F. serratus, which is the opposite order to that which would be expected from Keddy's model. Although all three species thrived well under the laboratory conditions, the results did not reflect in situ competitive dominances, which may be an effect of nutrient competition in the laboratory. Keddy's assumption that competitive ranks are consistent over the whole range of fundamental niches cannot be supported for Fucus spp. Communicated by K. Lüning     

Trichinella spiralis: intranasal immunization with attenuated Salmonella enterica carrying a gp43 antigen-derived 30mer epitope elicits protection in BALB/c mice

Trichinellosis is a public health problem and is considered an emergent/re-emergent disease in various countries. The etiological agent of trichinellosis is the nematode Trichinella, which infects domestic animals such as pigs and horses, as well as wild animals and humans. A veterinary vaccine could be an option to control the disease in domestic animals. Although several vaccine candidates have shown promising results, a vaccine against trichinellosis remains unavailable to date. Attenuated Salmonella strains are especially attractive live vectors because they elicit mucosal immunity, which is known to be important for the control of Trichinella spiralis infection at the intestinal level and can be administered by oral or intranasal routes. In this study, the autotransporter ShdA was used to display, on the surface of the Salmonella enterica serovar Typhimurium SL3261, the 210–239 amino acid epitope, (designated as Ag30) derived from the 43 kDa glycoprotein of T. spiralis muscle larvae. The fusion protein elicited antibodies in BALB/c mice that were able to recognize the native epitope on the surface of T. spiralis muscle larvae. Mice immunized by intranasal route with the recombinant Salmonella induced a protective immune response against the T. spiralis challenge, reducing by 61.83% the adult burden at day eight postinfection. This immune response was characterized by the induction of antigen-specific IgG1 and of IL-5 production. This study demonstrates the usefulness of Salmonella as a carrier of nematode epitopes providing a surface display system for intestinal parasite vaccine applications.     

Laboratory culture and taxonomy of two species ofPedobesia (Bryopsidales,Chlorophyceae) in Japan

The morphology ofPedobesia lamourouxii andDerbesia ryukyuensis, both collected in Shimoda and the adjacent areas in central Japan, was studied from field specimens and laboratory cultures. Specimens which had the same morphology as EuropeanP. lamourouxii produced stephanokont zoospores which developed into either prostrate filaments or expanded discoidal thalli similar to those described by Feldmann and Codomier (1974) and Feldmannet al. (1975). Erect filament identical with the thallus found in nature developed directly from prostrate filaments. The specimens which had morphology similar to that ofDerbesia ryukyuensis described by Yamada and Tanaka (1938) also produced stephanokont zoospores which developed similarly to those ofP. lamourouxii. This species is, therefore, a member ofPedobesia, and it is made a new combinationP. ryukyuensis (Yamada et Tanaka) Kobara et Chihara, comb. nov.     

The novel light-harvesting pigment-protein complex ofMantoniella squamata (Chlorophyta): Phylogenetic implications

Summary A light-harvesting pigment-protein complex has been isolated fromMantoniella squamata (Micromonadophyceae, Chlorophyta) by nondenaturing polyacrylamide-gel electrophoresis. The complex runs as two bands of molecular weights 54,000 and 55,000. There are two constituent polypeptides of molecular weights 20,500 and 22,000. Antibodies were raised to the 20,500-dalton polypeptides from this complex and to the 24,500-dalton polypeptide from the analogous complex ofPedinomonas minor (Micromonadophyceae). The antibodies to theM. squamata polypeptide are specific for both polypeptides of theM. squamata light-harvesting complex, as well as for a 27,000-dalton polypeptide of undetermined function. The antibodies to theP. minor polypeptide are specific for polypeptide components of the light-harvesting complex of that alga. The antibodies specific for theM. squamata light-harvesting complex polypeptides do not cross react with any polypeptides ofP. minor thylakoid membranes, as demonstrated by crossed immunoelectrophoresis. Similarly, no polypeptides ofM. squamata thylakoids cross react with the antibodies specific forP. minor light-harvesting complex polypeptides. These results indicate that the light-harvesting complex ofM. squamata is structurally very different from that ofP. minor. In a survey of several land plants and green algae, including representatives of all classes of green algae, a light-harvesting complex homologous to that ofM. squamata was found only inMicromonas pusilla. All other organisms tested possessed a lightharvesting complex homologous to that ofP. minor. The evolutionary and taxonomic implications of the novelM. squamata light-harvesting complex are discussed.     

The genetic analysis of F1 hybrid larvae between female Trichinella spiralis and male Trichinella britovi

Hybrids between female Trichinella spiralis and male Trichinella britovi were constructed. Then, hybrid genotype was characterized by DNA markers including mitochondrial cytochrome c oxidase subunit I (CO I) gene, the gene encoding the 43-kDa excretory–secretory (ES) protein, and genomic DNA fragments specific for T. spiralis and T. britovi identified from random amplified polymorphism DNA (RAPD). PCR-restriction fragment length polymorphism (PCR-RFLP) analysis of the mitochondrial CO I gene revealed that all hybrids carried a T. spiralis pattern. The same analysis of the gene encoding the 43-kDa ES protein showed that each hybrid carried both T. spiralis and T. britovi gene type simultaneously. In the analysis of genomic DNA using RAPD-derived PCR primers, some hybrids carried T. spiralis and T. britovi-specific RAPD markers, while others carried the RAPD marker of T. spiralis only.     

aubergine mutations in Drosophila melanogaster impair P cytotype determination by telomeric P elements inserted in heterochromatin

Transposable P elements inserted in the heterochromatic Telomeric Associated Sequences on the X chromosome (1A site) of Drosophila melanogaster have a very strong capacity to elicit the P cytotype, a maternally transmitted condition which represses P element transposition and P-induced hybrid dysgenesis. This repressive capacity has previously been shown to be sensitive to mutant alleles of the gene Su(var)205, which encodes HP1 (Heterochromatin Protein 1), thus suggesting a role for chromatin structure in repression. Since an interaction between heterochromatin formation and RNA interference has been reported in various organisms, we tested the effect of mutant alleles of aubergine, a gene that has been shown to play a role in RNA interference in Drosophila, on the repressive properties of telomeric P elements. Seven out of the eight mutant alleles tested clearly impaired the repressive capacities of the two independent telomeric P insertions at 1A analyzed. P repression by P strains whose repressive capacities are not linked to the presence of P copies at 1A were previously found to be insensitive to Su(var)205; here, we show that they are also insensitive to aubergine mutations. These results strongly suggest that both RNA interference and heterochromatin structure are involved in the establishment of the P cytotype elicited by telomeric P elements, and reinforce the hypothesis that different mechanisms for repression of P elements exist which depend on the chromosomal location of the regulatory copies of P.Communicated by G. Reuter     

ARE LIFE HISTORY CHARACTERISTICS GOOD PREDICTORS OF GENETIC DIVERSITY AND STRUCTURE? A CASE STUDY OF THE INTERTIDAL ALGA FUCUS SPIRALIS (HETEROKONTOPHYTA; PHAEOPHYCEAE)1

Characteristics of an organism's life history are often good predictors of genetic diversity and genetic structure. We tested hypotheses about genetic structure and diversity in an intertidal alga based on life history and life form. Fucus spiralis L. is a perennial monoecious alga that is abundant on the shores of Maine. Reproduction in fucoids is highly sensitive to water motion, resulting in fertilization success close to 100%. Given these life history characteristics, we predicted genetic structure among populations to be high and genetic diversity within populations to be low. We used five microsatellite loci to analyze genetic structure and diversity in F. spiralis from four sites on each of two coastal points, Maine, USA. Observed heterozygosities were relatively low (0.23 to 0.56), and F_IS estimates were usually significantly large, ranging from 0.021 to 0.476. This suggests that selfing and/or inbreeding may occur. Contrary to predictions, genetic differentiation between the two coastal points was insignificant. Moreover, few sites were genetically different from one another. Pairwise tests revealed complex patterns among sites. Genetic differentiation was not correlated with distance (P>0.05). Life history characteristics are good predictors of genetic diversity but not of population genetic structure in F. spiralis. We suggest that long distance dispersal of F. spiralis via drifting algal rafts increases gene flow. In addition, low levels of genetic structure may arise due to episodic recruitment or recent recolonization events. We discuss the implications of our results in terms of using life history characteristics as predictors of genetic diversity and structure in algae.     

Cloning and characterization of RAPD markers of parasitic nematodesTrichinella spiralis andT. pseudospiralis

A RAPD analysis was carried out in parasitic nematodesTrichinella spiralis andT. pseudospiralis, which are among the most common and hazardous agents in human and animal helminthoses. Several RAPD fragments were cloned and some of them sequenced. The fragments were shown to correspond to unique or low-repetitive sequences and to contain ORFs, microsatellites, and regions homologous to various pro- and eukaryotic sequences. Several fragments can be used to distinguish betweenT. spiralis andT. pseudospiralis.     

Trace elements found to be variable in two coral reef species,Heliofungia actiniformis andGalaxea fascicularis, collected from the ryukyu islands

Biominerals and metals of intertidal corals of two species (Heliofungia actiniformis, Quoy and Gaimard;Galaxea fascicularis, Linnaeus), collected from the Iriomote Island of Ryukyu, were examined with an inductively coupled plasma atomic emission spectrometer (ICP-AES). Twelve elements were detectable in the coralline skeletons dissected radially along the growth axis. The relative content (RC) of Hg periodically fluctuated and was minimum at the hollow sites of the coralline slab ofHeliofungia sp., corresponding to the cyclic growth. There were two types of elements: constant elements and variable elements along the growth axis. RCs of Ca, Mg, A1, Si, and P were nearly constant. RCs of Fe, Mn, Cu, and Ba were variable, but not as regularly changed as Hg. There were positive mass correlations of Hg to Mn, Cu and Zn, but not to Ba and Fe. In contrast, these relationships were not prominent and were likely degraded by aging in the skeleton ofGalaxea sp., suggesting a different mode from that of theHeliofungia sp.     

Water and solute regulation in Procephalothrix spiralis Coe and Clitellio arenarius (Müller) III. Long-term acclimation to diluted seawaters and effect of putative neuroendocrine structures

When acutely transferred to diluted seawater (SW), Procephalothrix spiralis and Clitellio arenarius regulate water content (g H₂O/g solute free dry wt = s.f.d.w.) via loss of Na and Cl (µmoles/g.s.f.d.w.). The present study extends these observations to a greater range of salinities and determines the effects of long-term, stepwise acclimation to diluted seawaters. Final exposure to a given experimental seawater (70, 50, 30, 15%) was 48 hours. Osmolality (mOsm/kg H₂O) and Na, K, and Cl ion concentrations (mEq/l) were determined in total tissue water and in the extracellular fluid of C. arenarius. Extracellular volume was determined as the ¹⁴C-polyethylene glycol space. Both species behaved as hyperosmotic conformers in diluted seawaters. However, reduction of the osmotic gradient between worm and medium occurred in P. spiralis, but not C. arenarius, in 30 and 15% SW. In both species, osmolality and Na, Cl, and K concentrations in total tissue water decreased with increased dilution of the SW. Water content increased with dilution of the medium but was lower than that which would be predicted based on approximation of the van't Hoff relation. This indicated the occurrence of regulatory volume decrease (RVD). In P. spiralis, in 70 or 50% SW, RVD was accompanied by loss of Na and Cl contents. However, in 30 or 15% SW, Na and Cl contents increased and in worms in 15% SW K content decreased. The latter movements of Na, Cl and K are indicative of cellular hysteresis and were associated with decreased viability, indicating the lower limits of regulatory ability in this species. In comparison, RVD in C. arenarius occurred in all diluted seawaters and was accompanied by loss of Na and Cl contents. In C. arenarius, evidence for reduced viability was absent. Removal of the supra- and subesophageal ganglia of C. arenarius resulted in retention of water, Na and Cl (g H₂O or µmoles/g s.f.d.w.) in worms acclimated to 70% SW. Removal of the cerebral ganglia and cephalic glands of P. spiralis did not significantly influence regulation of water content.     

Karyological and chemotaxonomical studies of the hybridPelargonium incarnatum ×P. patulum (Geraniaceae) and its parents

Pelargonium incarnatum is a tetraploid species (2n = 40) andP. patulum is diploid (2n = 22). A natural hybrid, collected at the same locality as the parent species, has 2n = 42 chromosomes. The meiosis of the hybrid is practically normal and its pollen fertility corresponds to that ofP. incarnatum. The flavonoid patterns of the parent species and the hybrid are very similar. The origin and phylogenetic implications of this intersectional hybrid are discussed.     

Competitive interactions between Fucus spiralis L. and F. vesiculosus L. (Fucales,Phaeophyta)

Fucus spiralis forms a conspicuous belt in the upper intertidal on rocky shores of the NW Atlantic. The objective of this study was to determine whether competition among congeners plays a role in restricting the distribution of F. spiralis to the upper shore. A replacement series design was used to test the growth performance of F. spiralis and F. vesiculosus in monocultures and in mixtures set up in the mid shore level on an exposed rocky coast. For F. spiralis and F. vesiculosus, all individual and relative crowding coefficients were < 1 and > 1, respectively. Hence, F. vesiculosus succeeded relative to F. spiralis in all mixtures tested. This finding was confirmed by graphical analysis. Ratio diagrams showed that the outcome of long term interaction among the species is extinction of F. spiralis. The results lead to the conclusion that interaction among congeners is a major determinant of the restricted distribution of F. spiralis.     

Trichinella spiralis: the influence of short chain fatty acids on the proliferation of lymphocytes, the goblet cell count and apoptosis in the mouse intestine

This study was carried out to determine the influence of short chain fatty acids (SCFA) on spleen and mesenteric lymph node lymphocyte proliferation, goblet cells and apoptosis in the mouse small intestine during invasion by Trichinella spiralis. BALB/c mice were infected with 250 larvae of T. spiralis. An SCFA water solution containing acetic, propionic and butyric acids (30:15:20 mM) was administered orally starting 5 days before infection and ending 20 days post infection (dpi). Fragments of the jejunum were collected by dissection 7 and 10 dpi, and were examined for apoptotic cells in the lamina propria of the intestinal mucosa, and for goblet cells. The proliferation index of the cultured spleen and mesenteric lymph node lymphocytes with MTT test was also determined. The orally administered SCFA solution decreased the proliferation of mesenteric lymph node lymphocytes in the mice infected with T. spiralis at both examination times, but did not influence the proliferative activity of the spleen cells. Seven dpi, both in the spleen and mesenteric lymph nodes, the highest proliferation index of concanavalin A (Con A)-stimulated lymphocytes was found in the group of uninfected animals receiving SCFA animals. This tendency could still be seen 10 dpi in the mesenteric lymph nodes but not in the spleen, where the proliferation index in this group had significantly decreased. In vitro studies revealed, that butyric and propionic acids added to the cell cultures suppressed the proliferation of Con A-stimulated mesenteric lymph nodes and spleen lymphocytes taken from uninfected and T. spiralis-infected mice. Acetic acid stimulated proliferation of splenocytes taken from uninfected mice but did not affect lymphocyte proliferation in mesenteric lymph nodes from uninfected or infected mice. Orally administered SCFA increased the number of goblet cells found in the epithelium of the jejunum 7 dpi, but this number had decreased 10 dpi. The number of apoptotic cells in the lamina propria of the intestinal mucosa of animals infected with the T. spiralis and receiving SCFA was also lower, particularly 10 dpi. The above results show that SCFA can participate in the immune response during the course of trichinellosis in mice.     

Studies of the genus Pityrosporum in submerged culture

Differentiation betweenPityrosporum ovale andPityrosporum orbiculare is mainly based upon the morphological features of cells and on physiological characters such as the ability ofP. ovale of utilizing oleic acid (Marples, 1965). Our studies in submerged culture, however, have not shown remarkable differences between the two species: growth curves, nutritional requirements, lipid utilization and optimal temperature are practically the same. In some strains of both species (1 : 4) we did observe a limited growth capacity, associated with a lower utilization rate of lipids. But for the notably different morphological features of cells,P. ovale andP. orbiculare could be ascribed, as regards their cultural characteristics, to the same species. The search forP. orbiculare was successful in all the cases of Pityriasis versicolor, while all our attempts to isolateP. ovale from its usual areas (scalp, upper back and chest) from both healty and seborrhoic subjets, gave no results.P. orbiculare was found in the same areas in the greatest part (85 %) of these subjects. In order to explain these results, one could assume thatP. orbiculare is the only species ofPityrosporum widespread in our region.These studies were supported by grant of the Consiglio Nazionale delle Ricerche of Italy under contract n. 70.01023/115.2798.     

Imazalil resistantPenicillium isolates from Spanish apple packinghouses

Imazalil tolerant isolates ofPenicillium spp. were recovered from sampling natural spore populations in storage rooms and apples collected from packinghouses in Lleida (Spain). ThePenicillium resistant strains belong to the speciesP. cyaneofulvum, P. variabile, P. rugulosum, P. minioluteum andP. pinophilum. 85% of the tested strains were resistant to imazalil in final concentrations of imazalil ranging from 4,500 µg/ml to 11,000 µg/ml. The resistance of these moulds to this fungicide was constant during successive subcultures. 89% ofPenicillium studied strains produced decay in the determination of parasitic fitness at 10 days.