Cell death triggering and effector recognition by Sw‐5 SD‐CNL proteins from resistant and susceptible tomato isolines to Tomato spotted wilt virus

Only a limited number of dominant resistance genes acting against plant viruses have been cloned, and further functional studies of these have been almost entirely limited to the resistance genes Rx against Potato virus X (PVX) and N against Tobacco mosaic virus (TMV). Recently, the cell‐to‐cell movement protein (NS_M) of Tomato spotted wilt virus (TSWV) has been identified as the avirulence determinant (Avr) of Sw‐5b‐mediated resistance, a dominant resistance gene which belongs to the class of SD‐CC‐NB‐LRR (Solanaceae domain‐coiled coil‐nucleotide‐binding‐leucine‐rich repeat, SD‐CNL) resistance genes. On transient expression of the NS_M protein in tomato and transgenic Nicotiana benthamiana harbouring the Sw‐5b gene, a hypersensitive cell death response (HR) is triggered. Here, it is shown that high accumulation of the Sw‐5b protein in N. benthamiana leaves, achieved by co‐expression of the Sw‐5b protein with RNA silencing suppressors (RSSs), leads to auto‐activity in the absence of NS_M. In a similar approach, Sw‐5a, the highest conserved paralogue of Sw‐5b from Solanum peruvianum, also triggered HR by auto‐activation, whereas the highest conserved orthologue from susceptible S. lycopersicum, named Sw‐5a^S, did not. However, neither of the last two homologues was able to trigger an NS_M‐dependent HR. Truncated and mutated versions of these Sw‐5 proteins revealed that the NB‐ARC [nucleotide‐binding adaptor shared by Apaf‐1 (from humans), R proteins and CED‐4 (from nematodes)] domain is sufficient for the triggering of HR and seems to be suppressed by the SD‐CC domain. Furthermore, a single mutation was sufficient to restore auto‐activity within the NB‐ARC domain of Sw‐5a^S. When the latter domain was fused to the Sw‐5b LRR domain, NS_M‐dependent HR triggering was regained, but not in the presence of its own Sw‐5a^S LRR domain. Expression analysis in planta revealed a nucleocytoplasmic localization pattern of Sw‐5b, in which the SD‐CC domain seems to be required for nuclear translocation. Although the Sw‐5 N‐terminal CC domain, in contrast with Rx, contains an additional SD, most findings from this study support a conserved role of domains within NB‐LRR (NLR) proteins against plant viruses.     

Comparative analysis of peroxiredoxin activation in the brown macroalgae Scytosiphon gracilis and Lessonia nigrescens (Phaeophyceae) under copper stress

Among thiol‐dependent peroxidases (TDPs) peroxiredoxins (PRXs) standout, since they are enzymes capable of reducing hydrogen peroxide, alkylhydroperoxides and peroxynitrite, and have been detected in a proteomic study of the copper‐tolerant species Scytosiphon gracilis. In order to determine the importance of these enzymes in copper‐stress tolerance, TDP activity and type II peroxiredoxin (II PRX) protein expression were compared between the opportunistic S. gracilis and the brown kelp Lessonia nigrescens, a species absent from copper‐impacted sites due to insufficient copper‐tolerance mechanisms. Individuals of both species were cultured with increasing copper concentrations (0–300 µg l^?1 Cu) for 96 h and TDP activity and lipoperoxides (LPXs) were determined together with II PRX expression by immunofluorescence and Western blot analysis. The results showed that TDP activity was higher in S. gracilis than L. nigrescens in all copper concentrations, independent of the reducing agent used (dithiothreitol, thioredoxin or glutaredoxin). This activity was copper inhibited in L. nigrescens at lower copper concentrations (20 µg l^?1 Cu) compared to S. gracilis (100 µg l^?1 Cu). The loss of activity coincided in both species with an increase in LPX, which suggests that TDP may control LPX production. Moreover, II PRX protein levels increased under copper stress only in S. gracilis. These results suggest that in S. gracilis TDP, particularly type II peroxiredoxin (II PRX), acts as an active antioxidant barrier attenuating the LPX levels generated by copper, which is not the case in L. nigrescens. Thus, from an ecological point of view these results help explaining the inability of L. nigrescens to flourish in copper‐enriched environments.     

<Emphasis Type="Italic">S</Emphasis>. Typhimurium <Emphasis Type="Italic">sseJ</Emphasis> gene decreases the <Emphasis Type="Italic">S</Emphasis>. Typhi cytotoxicity toward cultured epithelial cells

Background  

Salmonella enterica serovar Typhi and Typhimurium are closely related serovars as indicated by >96% DNA sequence identity between shared genes. Nevertheless, S. Typhi is a strictly human-specific pathogen causing a systemic disease, typhoid fever. In contrast, S. Typhimurium is a broad host range pathogen causing only a self-limited gastroenteritis in immunocompetent humans. We hypothesize that these differences have arisen because some genes are unique to each serovar either gained by horizontal gene transfer or by the loss of gene activity due to mutation, such as pseudogenes. S. Typhi has 5% of genes as pseudogenes, much more than S. Typhimurium which contains 1%. As a consequence, S. Typhi lacks several protein effectors implicated in invasion, proliferation and/or translocation by the type III secretion system that are fully functional proteins in S. Typhimurium. SseJ, one of these effectors, corresponds to an acyltransferase/lipase that participates in SCV biogenesis in human epithelial cell lines and is needed for full virulence of S. Typhimurium. In S. Typhi, sseJ is a pseudogene. Therefore, we suggest that sseJ inactivation in S. Typhi has an important role in the development of the systemic infection.     

First discovery of Stephanus Jurine (Hymenoptera: Stephanidae) in Japan,with description of a new species from Anijima Island of Ogasawara Islands

The genus Stephanus Jurine (in Panzer) is newly recorded from Japan and is represented by a new species. S. anijimensis sp. nov. from Anijima Island of the Ogasawara Islands. This species resembles S. tridentatus van Achterberg & Yang from the Oriental part of China, but can be distinguished by the combination of following characters: pronotum with a cavity under pronotal fold (cavity absent in S. tridentatus); hind femur comparatively slender, 4.8–5.1 times as long as maximum width (about 4.1 times in S. tridentatus); apical 0.5 of hind femur black or blackish‐brown (more or less yellowish in S. tridentatus). This new species was not found from other islands of the Ogasawara Islands, in which fauna is being severely endangered mainly due to predation by the introduced green anoles. Recently, it was introduced into Anijima Island in March 2013 and thus the extinction risk of this species is very high on this island.     

The relevance of life-history traits in the establishment of the invader <Emphasis Type="Italic">Eudiaptomus gracilis</Emphasis> and the extinction of <Emphasis Type="Italic">Eudiaptomus padanus</Emphasis> in Lake Candia (Northern Italy): evidence for competitive exclusion?

The niches invaded by exotic species are generally not entirely vacant, and one possible factor affecting the success of the invader to establish a permanent large population may be the ability of the former to outcompete native species. Eudiaptomus gracilis, which was not present in Italy before the 1980s, is becoming established in an increasing number of Northern Italian lakes, ultimately replacing the endemic E. padanus. Coexistence of the two species in Lake Candia lasted only 7 months, suggesting that species replacement was determined either by environmental changes or by strong competition. To assess whether the potential for interspecific competition existed and to identify species' traits which could explain the competitive superiority of E. gracilis, we examined field seasonal patterns, reproductive parameters and body size of the two species. E. padanus abundance was probably reduced by predation and parasitism, which favoured the displacement of the species shortly after invasion by E. gracilis. The temporarily underexploited niche provided an opportunity for the successful establishment of the invader. The reproductive patterns of the two species were found to be similar in most of their features, with the exception of a markedly larger clutch size and a smaller egg volume in the resident species. By contrast, the invader showed a higher adult:egg ratio and a lower death rate. Thus, despite the greater fecundity of E. padanus, the competitive success of the invader might be attributable to interspecific differences in developmental rates and/or juvenile mortality.     

The C‐type lectin receptor MGL senses N‐acetylgalactosamine on the unique Staphylococcus aureus ST395 wall teichoic acid

Staphylococcus aureus is a common skin commensal but is also associated with various skin and soft tissue pathologies. Upon invasion, S. aureus is detected by resident innate immune cells through pattern‐recognition receptors (PRRs), although a comprehensive understanding of the specific molecular interactions is lacking. Recently, we demonstrated that the PRR langerin (CD207) on epidermal Langerhans cells senses the conserved β‐1,4‐linked N‐acetylglucosamine (GlcNAc) modification on S. aureus wall teichoic acid (WTA), thereby increasing skin inflammation. Interestingly, the S. aureus ST395 lineage as well as certain species of coagulase‐negative staphylococci (CoNS) produce a structurally different WTA molecule, consisting of poly‐glycerolphosphate with α‐O‐N‐acetylgalactosamine (GalNAc) residues, which are attached by the glycosyltransferase TagN. Here, we demonstrate that S. aureus ST395 strains interact with the human Macrophage galactose‐type lectin (MGL; CD301) receptor, which is expressed by dendritic cells and macrophages in the dermis. MGL bound S. aureus ST395 in a tagN‐ and GalNAc‐dependent manner but did not interact with different tagN‐positive CoNS species. However, heterologous expression of Staphylococcus lugdunensis tagN in S. aureus conferred phage infection and MGL binding, confirming the role of this CoNS enzyme as GalNAc‐transferase. Functionally, the detection of GalNAc on S. aureus ST395 WTA by human monocyte‐derived dendritic cells significantly enhanced cytokine production. Together, our findings highlight differential recognition of S. aureus glycoprofiles by specific human innate receptors, which may affect downstream adaptive immune responses and pathogen clearance.     

Non-monophyly of fish in the Sinipercidae (Perciformes) as inferred from cytochrome <Emphasis Type="Italic">b</Emphasis> gene

The sinipercids represent a group of 12 species of freshwater percoid fish, including nine in Siniperca and three species in Coreoperca. Despite several classification attempts and a preliminary molecular phylogeny, the phylogenetic relationships and systematic position of sinipercids remained still unsolved. The complete cytochrome b gene sequences from nine sinipercid species four non-sinipercid fish species were cloned, and a total of 12 cyt b sequences from 10 species of sinipercids and 11 cyt b sequences from 10 species of non-sinipercid fish also in Perciformes were included in the phylogenetic analysis. As expected, the two genera Siniperca and Coreoperca within sinipercids are recovered as monophyletic. However, nine species representing Moronidae, Serranidae, Centropomidae, Acropomatidae, Emmelichtyidae, Siganidae and Centrarchidae included in the present study are all nested between Coreoperca and Siniperca, which provides marked evidence for a non-monophyly of sinipercid fishes. Coreoperca appears to be closest to Centrachus representing the family Centrarchidae. Coreoperca whiteheadi and C. herzi are sibling species, which together are closely related to C. kawamebari. In the Siniperca, the node between S. roulei and the remaining species is the most ancestral, followed by that of S. fortis. S. chuatsi and S. kneri are sibling species, sister to S. obscura. However, the sinipercids do not seem to have a very clear phylogenetic history, for different methods of phylogenetic reconstruction result in different tree topologies, and the only conclusive result in favor of a paraphyletic origin of the two sinipercid genera is the parametric bootstrap test. The paraphyly of Sinipercidae may suggest that the “synapomorphs” such as cycloid scales, upon which this family is based, were independently derived at least twice within sinipercid fishes, and further study should be carried out to include the other two Siniperca species and to incorporate other genes. Handling editor: C. Sturmbauer     

Evidence of hybridization in the Argentinean lizards Liolaemus gracilis and Liolaemus bibronii (IGUANIA: LIOLAEMINI): an integrative approach based on genes and morphology

The lizard genus Liolaemus is endemic to temperate South America and includes more than 225 species. Liolaemus gracilis and L. bibronii are closely related species that have large and overlapping geographic distributions, and the objective of this work is to further investigate the L. bibronii–L. gracilis mtDNA paraphyletic pattern previously detected, using an integrative approach, based on mtDNA, nuclear DNA and morphological characters. We identified eight morphological L. bibronii introgressed with L. gracilis mtDNAs, and the reciprocal for one L. gracilis, from six localities in the region of sympatry overlap. The morphological identity of these introgressed individuals was confirmed by diagnostic nuclear markers, and this represents the first well-documented case of interspecific hybridization in the lizard genus Liolaemus. Of the three most likely hypotheses for these observed patterns, we suggest that asymmetrical mtDNA introgression as a result of recent or ongoing hybridization between L. bibronii and L. gracilis is the most likely. This may be due to size selection by L. gracilis female preference for the larger L. bibroni males in sympatry, but this requires experimental confirmation.     

GRACILARIA HUMMII SP. NOV. (GRACILARIALES,RHODOPHYTA), A NEW NAME FOR THE AGAROPHYTE “GRACILARIA CONFERVOIDES” HARVESTED IN NORTH CAROLINA DURING WORLD WAR II1

Gracilaria hummii Hommers. et Freshwater is proposed as a new name for the inshore cylindrical species found in North Carolina that was treated as Gracilaria confervoides (L.) Grev. during World War II, and more recently as G. verrucosa (Huds.) Papenf. Molecular evidence places G. hummii in the Gracilis‐group in Gracilaria together with G. gracilis (Stackh.) Steentoft, L. M. Irvine et Farnham, the name currently applied to specimens formerly identified as G. confervoides and G. verrucosa. G. hummii differs from G. gracilis in possessing shallower male conceptacles in which the spermatangial filament originates from a surface cortical cell rather than from a subcortical cell. The cystocarps are similar, except that the gonimoblasts of G. hummii are attached to the base of the pericarp by numerous, prominent thickened terminal tubular cells and because terminal tubular cells are absent above the midregion of the cystocarp or in the vicinity of the ostiole. The gonimoblasts are subtended by a bundle of longitudinally oriented, thick‐walled secondary filaments of a type that has not been described before in Gracilaria but that may be characteristic of some other species. G. hummii occupies a basal position in the Gracilis‐group and is distinct from all other cylindrical North Carolina Gracilaria species, according to the molecular and morphological evidence.     

The Tomato spotted wilt virus cell‐to‐cell movement protein (NSM) triggers a hypersensitive response in Sw‐5‐containing resistant tomato lines and in Nicotiana benthamiana transformed with the functional Sw‐5b resistance gene copy

Although the Sw‐5 gene cluster has been cloned, and Sw‐5b has been identified as the functional gene copy that confers resistance to Tomato spotted wilt virus (TSWV), its avirulence (Avr) determinant has not been identified to date. Nicotiana tabacum ‘SR1‘ plants transformed with a copy of the Sw‐5b gene are immune without producing a clear visual response on challenge with TSWV, whereas it is shown here that N. benthamiana transformed with Sw‐5b gives a rapid and conspicuous hypersensitive response (HR). Using these plants, from all structural and non‐structural TSWV proteins tested, the TSWV cell‐to‐cell movement protein (NS_M) was confirmed as the Avr determinant using a Potato virus X (PVX) replicon or a non‐replicative pEAQ‐HT expression vector system. HR was induced in Sw‐5b‐transgenic N. benthamiana as well as in resistant near‐isogenic tomato lines after agroinfiltration with a functional cell‐to‐cell movement protein (NS_M) from a resistance‐inducing (RI) TSWV strain (BR‐01), but not with NS_M from a Sw‐5 resistance‐breaking (RB) strain (GRAU). This is the first biological demonstration that Sw‐5‐mediated resistance is triggered by the TSWV NS_M cell‐to‐cell movement protein.     

Changes in a Sudano-Sahelian rodent community after slash-and-burn farming (Gonsé Forest, Burkina Faso)

This study describes the recolonization by a Sudano‐Sahelian rodent community of an area slashed and burned in the middle of the dry season (Gonsé Forest in Burkina Faso). A series of abundance cycles occurred. The first began with the wet season and the arrival of a succession of four species: Taterillus gracilis, Tatera guineae, Nannomys sp. and Mastomys erythroleucus. Taterillus gracilis appeared to be the least demanding species. It was sexually active whatever the season and took advantage of reduced competition among species on the burned tracts. Tatera guineae also extended its spatial distribution but failed to reproduce. Nannomys sp. rapidly increased after the first rains. Although generally held to be the most prolific and opportunistic of the species involved, M. erythroleucus only found conditions amenable to its colonization late on. The timing of the disruption and whether or not it coincides with the reproductive period and the mobility phase of each species are decisive factors. This information is useful for a clearer understanding of the dynamics of each species and for evaluating the risk and probable locations of outbreaks of certain species.     

Evidence for two sympatric species of snipefishes<Emphasis Type="Italic"> Macroramphosus</Emphasis> spp. (Syngnathiformes,Centriscidae) on Great Meteor Seamount

About 202 specimens of snipefishes (Macroramphosus spp.) from Great Meteor Seamount (GMR, subtropical NE Atlantic, 30°N, 28.5°W) were analysed with respect to diet composition and morphology. Fifty specimens belonged to the deep-bodied benthos-feeding type (b-type) whose diet consisted of foraminifers, pteropods, decapods and polychaetes, whereas the slender planktivorous individuals (p-type, n=140) mainly fed on ostracods, copepods, pteropods and foraminifers. Twelve specimens showed no specialisation with respect to feeding (p/b-type). Both feeding types can be significantly distinguished from each other by means of bi- and multivariate morphological analysis considering the variables body depth, length of second dorsal spine, diameter of orbit and standard length. We discuss the hypothesis that M. gracilis represents a transient juvenile stage of M. scolopax. Since our specimens of the M. gracilis type were larger than specimens of the M. scolopax-type, such an ontogenetic shift is unlikely to occur. Our results support the hypothesis of Clarke for Australian snipefishes that for Macroramphosus spp. locally two distinct sympatric species must be anticipated, corresponding to M. scolopax and M. gracilis and the b- and p-types, respectively. Electronic Publication     

Checklist of the freshwater fishes of Armenia,Azerbaijan and Georgia

We present a critical checklist of freshwater fish species found so far in the countries of Armenia, Azerbaijan and Georgia. In total 119 freshwater fishes are recorded. There are 40, 86 and 96 species currently known for Armenia, Azerbaijan and Georgia respectively. From these 119 species, seven are endemic and seven species are alien. From the alien species, only three (Carassius gibelio, Gambusia holbrooki and Pseudorasbora parva) can be considered as widespread and invasive. There are four species (Gasterosteus aculeatus, Gobio artvinicus, Perca fluviatilis and Salmo gegarkuni) that are translocated within the region. Seven species are confirmed or recorded for the first time including G. artvinicus and Oxynoemacheilus veyselorum for Armenia, Azerbaijan and Georgia, Capoeta kaput and Rhinogobius lindbergi for Azerbaijan and Georgia, Capoeta razii for Azebaijan, Oxynoemacheilus cemali and Squalius agdamicus for Georgia. In this checklist, Acipenser colchicus is treated as a synonym of Acipenser persicus. Sand smelts of the Black and Caspian Sea basin are identified as Atherina caspia and Clupeonella caspia is treated as a synonym of Clupeonella cultriventris. Coregonus sevanicus is listed as Coregonus sp. until the situation of Sevan whitefish is better understood. Capoeta sevangi and Capoeta ekmekciae are synonyms of Capoeta capoeta. The fish often identified as Capoeta capoeta gracilis from rivers south of the Kura most likely belong to C. razii. The Black and Caspian Sea Rutilus populations are treated as conspecific, therefore R. kutum is a junior synonym of R. frisii. Oxynoemacheilus veyseli is valid as O. veyselorum. We list the alien Rhinogobius species as R. lindbergi, however the name is provisional and needs further confirmation. All Squalius species from the Kura River drainage are identified as S. agdamicus, however in the Aras, it is replaced by S. turcicus. Squalius orientalis is treated as a valid species restricted to the eastern Black Sea basin. The four forms of Lake Sevan trout (Salmo ischchan, S. gegarkuni, S. danilewskii and S. aestivalis) are treated as valid species, two of them (S. ischchan and S. danilewskii) are extinct. Rutilus sojuchbulagi from Azerbaijan is also extinct.     

<Emphasis Type="Italic">Diplectanocotyla</Emphasis> Yamaguti, 1953 (Monogenea: Diplectanoidea) from <Emphasis Type="Italic">Megalops cyprinoides</Emphasis> (Broussonet) (Teleostei: Megalopidae) off Peninsular Malaysia

Two known and two new species of Diplectanocotyla Yamaguti, 1953 (D. gracilis Yamaguti, 1953, D. megalopis Rakotofiringa & Oliver, 1987, D. langkawiensis n. sp. and D. parva n. sp.) were collected from Megalops cyprinoides (Megalopidae) off Langkawi, Kedah and Matang, Perak, Peninsular Malaysia. All four species possess similar types of sclerotised male and female reproductive structures and similar soft anatomical features. The squamodisc sclerites of all four species have spine-like projections with varying degrees of visibility and shapes (sharp-pointed to triangular). In D. megalopis and D. langkawiensis n. sp. the spines are sharp-pointed and distinct on sclerites from rows 5–6 onwards. In D. gracilis and D. parva n. sp. the sclerite spines are triangular, lightly sclerotised and occur on almost all of the sclerites. D. parva n. sp. has comparatively the smallest set of anchors, bars, squamodiscs and squamodisc suckers. The anchors and bars of the other three species are almost similar in overall size, and the main distinguishing feature is the relative lengths of the inner and outer roots of the ventral anchors. In D. gracilis the outer root is very much smaller than the inner root and they are disposed almost at a right angle to each other. In D. megalopis the outer root is usually about half the length of the inner root and the roots are inclined at c.60° to each other. In D. langkawiensis n. sp. the roots are inclined at c.40° degrees and the outer root is of a similar length or only slightly shorter than the inner root. The openings of the two squamodisc suckers of all four Diplectanocotyla species are surrounded by tiny scale-like spines. Bifid tegumental spines are found in the posterior region of all four species, differing only in their extent: in D. parva n. sp. the tegumental spines are only distributed in the peduncular region and not beyond, whilst in the other three species the tegumental spines extend from the posterior level of the testis to the end of the peduncle. An amended diagnosis of Diplectanocotyla and a key to its species are appended.     

Callus induction and regeneration in<Emphasis Type="Italic"> Spirodela</Emphasis> and<Emphasis Type="Italic"> Lemna</Emphasis>

The development of tissue culture systems in duckweeds has, to date, been limited to species of the genus Lemna. We report here the establishment of an efficient tissue culture cycle (callus induction, callus growth and plant regeneration) for Spirodela oligorrhiza Hegelm SP, Spirodela punctata 8717 and Lemna gibba var. Hurfeish. Significant differences were found among the three duckweed species pertaining to carbohydrate and phytohormone requirements for callus induction, callus growth and frond regeneration. In vitro incubation with poorly assimilated carbohydrates such as galactose (S. oligorrhiza SP and L. gibba var. Hurfeish) and sorbitol (S. punctata 8717) as sole carbon source yielded high levels of callus induction on phytohormone-supplemented medium. Sorbitol is required for optimal callus growth of S. oligorrhiza SP and S. punctata 8717, while sucrose is required for callus growth of L. gibba var. Hurfeish. Sucrose either alone (S. oligorrhiza SP, L. gibba var. Hurfeish) or in addition to sorbitol (S. punctata 8717) is required for frond regeneration.Abbreviations ABA: (±)-Abscisic acid - BA: N⁶-Benzyladenine - 2,4-D 2,4-Dichlorophenoxyacetic acid - Dicamba: 3,6-Dichloro-2-methoxybenzoic acid - 2iP: N⁶-(2-Isopentenyl)adenine - NAA: -Naphthaleneacetic acid - PCA: p-Chlorophenoxy acetic acid - Picloram: 4-Amino-3,5,6-trichloropicolinic acid - TDZ: Thidiazuron Communicated by A. AltmanJ. Li and M. Jain contributed equally to the research reported in this article.     

Multivariate analyses of morphometrical features from Apatemon gracilis (Rudolphi, 1819) Szidat, 1928 and A. annuligerum (v. Nordmann, 1832) (Digenea: Strigeidae) metacercariae

Only two species of the strigeid subgenus Apatemon (Apatemon) sensu Sudarikov (1959), Apatemon gracilis (Rudolhi, 1819) and A. annuligerum (von Nordmann, 1832), are known to infect British fishes. A. annuligerum is poorly characterised, with few life-cycle data available, since only the adult and metacercarial stages have been described. Both of these life-stages demonstrate only minor morphological differences from those of A. gracilis. The multidimensional approach of principal components analysis (PCA) was employed to determine the intra- and inter-specific morphological variation that exists for metacercariae of these species. Analyses revealed that metacercarial morphometrics cannot confidently discriminate between Apatemon gracilis and A. annuligerum specimens. Indeed, levels of intra-specific variation present between A. gracilis metacercariae originating from different piscine hosts were as marked for certain populations as those between the two nominal species. Individual populations of A. gracilis metacercariae differed most in the relative dimensions of their internal organs, whereas variation between the two nominal species was primarily in length-related variables. A. gracilis specimens originating from Welsh and Scottish stoneloach represented two populations with the most comparable total body lengths, yet clustered separately. Conversely, A. gracilis metacercariae infecting rainbow trout and salmon parr differed markedly in size, but demonstrated minimal composite morphological variation. Much, if not all, of the variation described between populations and species of parasites can be attributed to host or to metacercarial location within a particular host. These findings and the lack of strong life-cycle data for A. annuligerum place some doubt on its validity as a species discrete from A. gracilis.     

SopB‐ and SifA‐dependent shaping of the Salmonella‐containing vacuole proteome in the social amoeba Dictyostelium discoideum

The ability of Salmonella to survive and replicate within mammalian host cells involves the generation of a membranous compartment known as the Salmonella‐containing vacuole (SCV). Salmonella employs a number of effector proteins that are injected into host cells for SCV formation using its type‐3 secretion systems encoded in SPI‐1 and SPI‐2 (T3SS‐1 and T3SS‐2, respectively). Recently, we reported that S. Typhimurium requires T3SS‐1 and T3SS‐2 to survive in the model amoeba Dictyostelium discoideum. Despite these findings, the involved effector proteins have not been identified yet. Therefore, we evaluated the role of two major S. Typhimurium effectors SopB and SifA during D. discoideum intracellular niche formation. First, we established that S. Typhimurium resides in a vacuolar compartment within D. discoideum. Next, we isolated SCVs from amoebae infected with wild type or the ΔsopB and ΔsifA mutant strains of S. Typhimurium, and we characterised the composition of this compartment by quantitative proteomics. This comparative analysis suggests that S. Typhimurium requires SopB and SifA to modify the SCV proteome in order to generate a suitable intracellular niche in D. discoideum. Accordingly, we observed that SopB and SifA are needed for intracellular survival of S. Typhimurium in this organism. Thus, our results provide insight into the mechanisms employed by Salmonella to survive intracellularly in phagocytic amoebae.     

A preliminary investigation on the growth requirement for monovalent cations, divalent cations and medium ionic strength of marine actinomycete Salinispora

In this paper, we report that three species of Salinispora, S. arenicola, S. tropica, and S. pacifica, require magnesium and calcium, for growth, with S. pacifica having the most stringent growth requirement for these ions. Interaction between these ions in supporting the growth of Salinispora was observed. We also demonstrated that the absolute requirement of sodium to support the growth of Salinispora has not been established as all three species of Salinispora can use either potassium or lithium to replace sodium to support maximum growth. While lithium can replace sodium to support maximum growth of Salinispora, it is more toxic to S. arenicola than S. tropica and S. pacifica, inhibiting the growth of S. arenicola at 189 mM but without effect on the growth of S. tropica and S. pacifica. Using both sodium chloride-based and lithium chloride-based media, we showed that Salinispora has a growth requirement for divalent ions, magnesium and calcium as well as growth requirement for ionic strength (8.29 to 15.2 mS/cm). S. arenicola has a lower growth requirement for ionic strength than S. tropica and S. pacifica.