Taxonomic review and key to European Ichneumon flies (Hymenoptera, Ichneumonidae), parasitoids of gall-forming sawflies of the genera Pontania Costa, Phyllocolpa Benson, and Euura Newman (Hymenoptera, Tenthredinidae) on willows: Part I

Extensive material on parasitoids of the family Ichneumonidae reared from gall-making sawflies on Salix is identified. The general host—parasite list includes 75 species of hosts—Pontania (40 spp.), Phyllocolpa (21 spp.), Euura (14 spp.), their food plants (33 species of Salix), and their parasitoids of the family Ichneumonidae; many data on hosts and parasitoids are recorded for the first time. The complex of ichneumonids on gallforming sawflies is composed of about 45 species belonging to 15 genera of 6 subfamilies. The annotated list of about 30 species of ichneumonids (except for species of Saotis) reared from gall-making sawflies is given with data on the literature, studied material, distribution, and hosts. Two new species Anoncus gallicola sp. n. and Rhinotorus brachycerus sp. n. are described. A key to the species of the Ichneumonidae of this complex is given.     

Bipolar heterothallism,a principal mating system of<Emphasis Type="Italic">Cordyceps militaris in vitro</Emphasis>

Interest inin vitro study of entomopathogenic fungi, includingCordyceps species, has been increasing due to their valuable bioactive compounds and biocontrol effects. AmongCordyceps species,in vitro stromata ofC. militaris has been successfully produced and cultivated for industrial purposes. However, genetic study onin vitro stromata formation ofC. militaris has not been carried out yet. Here, relationship between mating system and perithecial stromata formation ofC. militaris is reported. Mating system was determined by observing perithecial stromata formation from mono-ascospore cultures and their pair-wise combinations. Certain combinations of mono-ascospore strains produced perithecial club-shaped stromata, whereas other combinations produced either no stromata or only abnormal non-perithecial stromata. Similarly, monoascospore cultures without combination produced either no stromata or only abnormal nonperithecial stromata. Despite obvious heterothallism, self-fertility was occasionally observed in few strains ofC. militaris. These observations indicated thatC. militaris behaves as a bipolar heterothallic fungus and requires two mating compatible strains in order to produce regular clubshaped perithecial stromata, a fundamental requirement for its industrial cultivation.     

Molecular phylogeny of the entomopathogenic fungi of the genus Cordyceps (Ascomycota: Clavicipitaceae) and its evolutionary implications

Abstract Cordyceps is an endoparasite ascomycetous genus containing approximately 450 species with a diversity of insect hosts, traditionally included in the family Clavicipitaceae of Ascomycota. Establishing the relationships among species with a varied range of morphologies and hosts is of importance to our understanding of the phylogeny and co‐evolution of parasites and hosts in entomopathogenic ascomycetes. To this end, we used a combination of molecular index and morphological characters from 40 representative species to carry out comprehensive molecular phylogenetic analyses. Based on the phylogenetic tree, we used the program DISCRETE for inferring the rates of evolution and finding ancestral states of morphological character. The phylogenetic analyses revealed two important points. (i) Types of perithecia attached to stroma reflected an evolutionary trend in Cordyceps. The vertically immersed perithecia form was the ancestral state, superficial and obliquely immersed perithecia were derived characters, obliquely immersed was irreversible. Species with obliquely immersed perithecia were in a closely related group and were the derived group. (ii) A strong correlation between fungal relatedness and the microhabitat supported the hypothesis that the host jumps through commingling in soil microhabitats. Based on the results of these analyses, host switching explains the diversity of entomopathogenic fungi of the genus Cordyceps.     

A taxonomic review of ichneumon-flies (Hymenoptera,Ichneumonidae), parasitoids of gall-forming sawflies (Hymenoptera,Tenthredinidae) on Salix. Part II. Review of the Palaearctic species of the genus <Emphasis Type="Italic">Saotis</Emphasis> Förster with description of four new species

This is the second communication on Ichneumonidae, parasitoids of gall-making sawflies on Salix. Twenty-one Palaearctic species of the specialized genus Saotis are revised and classified into 12 species groups. Original data on 29 species of Saotis hosts are given: 18 spp. of Phyllocolpa, 10 Pontania and 1 species of Pontopristia. A key to species and subspecies is given and illustrated by 55 figures. For each species the literature, synonymy, diagnosis, material and hosts are given. Four new species are described: Saotis alpinator sp. n., S. boreator sp. n., S. granulator sp. n. and S. subarctor sp. n.     

Dispersal flight and colony development in the fungus-growing termites <Emphasis Type="Italic">Pseudacanthotermes spiniger</Emphasis> and <Emphasis Type="Italic">P. militaris</Emphasis>

Pseudacanthotermes spiniger and P. militaris are two African fungus-growing termites (Termitidae, Macrotermitinae) which may become pests in disturbed agrosystems where they often live in sympatry. To study their development and their reproductive strategies, colonies of both species were reared in the laboratory for 20 and 17 years, respectively, after their foundation from reproductive pairs. The first steps of development were in great part similar in both species, although P. spiniger favoured the defence during the juvenile period, while P. militaris tended to favour a rapid development. While P. spiniger colonies did not produce alates until year 7 of colony life, P. militaris colonies were able to produce a fertile progeny 4 years after their foundation. In contrast, major soldiers were more rapidly differentiated in the incipient colonies of P. spiniger. Dispersal flights occurred every year for 10 years in P. spiniger. In P. militaris, dispersal flights did not occur regularly although alates appeared yearly. The annual number of alates produced by P. spiniger increased with the colony age to reach a maximum of 25,000 individuals and global production of alates was estimated at ca. 150,000 individuals in the life of a colony. The longevity of P. spiniger and P. militaris colonies was around 20 years. These species were shown to be reproductively isolated by multiple pre-mating mechanisms. While chronological differences in dispersal flights contribute to reproductive isolation of the two species, the non-viability of experimental hybrid colonies also indicates the involvement of post-mating mechanisms of isolation.     

Preservation affects the vegetative growth and fruiting body production of <Emphasis Type="Italic">Cordyceps militaris</Emphasis>

Cordyceps militaris is a model species of Cordyceps fungi, and has been traditionally used as an edible and medicinal fungus due to its richness of bioactive and pharmacological metabolites. The fruiting bodies of this fungus are widely used as healthy food and nutrition supply. In industrial production, fruiting bodies are cultivated on artificial media, but their yield and quality are usually affected by the quality of fungal strains. In this study, the effect of colony growth rate of the fungal strains, fungal age and repeated subculturing on the fungal biomass accumulation was investigated. The results indicated that the fungal biomass was positively correlated with the colony growth rate and not affected by fungal age and the repeated subculturing. The preservation conditions for stock cultures, including choice of cultures, lyophilization, temperature and protective agents were optimized based on the mycelial formation and conidia production in artificial inoculum. The development of fruiting bodies from the fungal strains stored under the optimized preservation conditions were further analyzed to determine the ideal time period of preservation. Results indicated that storing the fungus at 4 °C could maintain the fungal vitality and fruiting body producing capacity for at least 12 months. This study established practical criteria of fungal inoculum for artificial cultivation of fruiting body and provided a simple and efficient preservation method for C. militaris. The results may shed light on preservation for other Cordyceps species and other edible fungi.     

<Emphasis Type="Italic">Metarhizium taii var. chongqingensis</Emphasis> Nov., Anamorph of <Emphasis Type="Italic">Cordyceps</Emphasis><Emphasis Type="Italic">chongqingensis</Emphasis> sp. Nov. Isolated from a Low Altitude Area in Chongqing,China

Previously, a new Cordyceps species was isolated from a low altitude area in Chongqing, China, and named Cordyceps chongqingensis sp. nov. In this study, its anamorph was isolated and designated CQM1^T. It grew optimally on Czapek medium supplied with 0.5% silkworm flour and 0.5% soybean oil meal at 25°C, pH 5.0–5.5. The phenotypic and molecular characteristics were investigated for its identification and typing. Morphological observations under a microscope revealed that this anamorph of Cordyceps chongqingensis sp. nov. was a new species of Metarhizium. Moreover, it was identified as one of the variants of Metarhizium taii based on sequences of 26S rDNA D1/D2 and ITS regions, and thus named Metarhizium taii var. chongqingensis nov.     

Host specificity and host recognition in a chemically-defended herbivore, the tenthredinid sawfly Rhadinoceraea nodicornis

The sawfly Rhadinoceraea nodicornis Konow (Hymenoptera: Tenthredinidae) is a member of a closely related group of species, the tribe Phymatocerini, which feed on the Liliales and Ranunculales. It is known to sequester steroid alkaloids from its host plants, species in the genus Veratrum (Liliales: Melanthiaceae), and to use them as a defence against predators. There are known chemical relationships between the hosts of R. nodicornis and hosts of related sawfly species. We tested whether the R. nodicornis larvae would accept hosts of closely- and more distantly-related sawflies, but found that they accepted only plant species in the genus Veratrum. This specificity was apparently innate, as it was independent of early larval experience. A feeding bioassay showed that the steroid alkaloids from Veratrum nigrum were phagostimulatory for R. nodicornis larvae, suggesting that they may be involved in host recognition. We discuss the possibility that the evolution of recognition of specific compounds may represent the mechanism of host radiation within the Phymatocerini.     

Beiträge zur Kenntnis der europäischen Raupenfliegen (Dipt. Tachinidae)

Summary In this revision of the European species of the genusMyxexoristops t. t. (parasites of sawflies) six species, including a new one (abietis n. sp., parasite ofCephalcia), are described by their distinguishing characters. Separate keys for the determination of males and females are given. For five species, hosts are recorded on the basis of bred material examined by the author himself.Zenillia nox hall, described from Japan is identical to the European speciesM. stolida stein (nov. syn.).      

Two new species of <Emphasis Type="Italic">Cotingacola</Emphasis> Carriker,Phthiraptera: Ischnocera: Philopteridae) from Amazonian Brazil,with comments on host-specificity

This paper describes two new species of Cotingacola Carriker, 1956, C. lutzae n. sp. and C. engeli n. sp., from the cinereous mourner Laniocera hypopyrra (Vieillot) and black-necked red cotinga Phoenicircus nigricollis Swainson (Aves: Passeriformes: Cotingidae), respectively. These species are fully illustrated and compared with their morphologically closest relatives. With the addition of these two new species, this Neotropical chewing louse genus now includes ten species. We also present records from Brazil for two previously described species, C. stotzi Clayton & Price, 1998 and C. parmipapillae Carriker, 1956. The re-examination of a specimen identified as ‘Cotingacola sp.’ in a recent phylogenetic study confirmed that it is C. parmipapillae and that Querula purpurata (Statius Müller) regularly hosts two Cotingacola species, which means that at least three cotingid hosts carry more than one species of Cotingacola. We comment on the variability in the host-specificity of Cotingacola species.     

Variations of SSU rDNA group I introns in different isolates of Cordyceps militaris and the loss of an intron during cross-mating

Cordyceps militaris, the type species of genus Cordyceps, is one of the most popular mushrooms and a nutraceutical in eastern Asia. It is considered a model organism for the study of Cordyceps species because it can complete its life cycle when cultured in vitro. In the present study, the occurrence and sequence variation of SSU rDNA group I introns, Cmi.S943 and Cmi.S1199, among different isolates of C. militaris were analyzed. Based on the secondary structure predictions, the Cmi.S943 intron has been placed in subgroup IC1, and the Cmi.S1199 intron has been placed in subgroup IE. No significant similarity between Cmi.S943 and Cmi.S1199 suggested different origins. Three genotypes, based on the frequency and distribution of introns, were described to discriminate the 57 surveyed C. militaris strains. It was found that the genotype was related to the stroma characteristics. The stromata of all of the genotype II strains, which possessed only Cmi.S943, could produce perithecium. In contrast, the stromata of all genotype III strains, which had both Cmi.S943 and Cmi.S1199, could not produce perithecium. Cmi.S1199 showed the lowest level of intra-specific variation among the tested strains. Group I introns can be lost during strain cross-mating. Therefore, we presumed that during cross-mating and recombination, intron loss could be driven by positive Darwinian selection due to the energetic cost of transcribing long introns.     

Morphology and phylogenetic position of two novel marine gregarines (Apicomplexa,Eugregarinorida) from the intestines of North‐eastern Pacific ascidians

Eugregarine apicomplexans parasitize marine, freshwater and terrestrial invertebrates, and have lifecycles involving trophozoites (feeding stages) with complex morphologies and behaviour. The genus Lankesteria refers to marine aseptate eugregarines that parasitize ascidians. We described the surface ultrastructure of two new gregarine species, L. chelyosomae sp. n. and L. cystodytae sp. n. that inhabit the intestines of Chelyosoma columbianum and Cystodytes lobatus, respectively, collected from the North‐eastern Pacific Ocean. Apart from inhabiting different hosts and major differences in the cell size of L. chelyosomae sp. n. (mean length 182 µm) and L. cystodytae sp. n. (mean length 70 µm), the morphology of both gregarine species was quite similar. The trophozoites ranged from elliptoid to obdeltoid in shape and were brownish in colour. The nucleus was situated at the anterior end of the cell just behind a pointed mucron. A dense array of epicytic knobs was present over the entire surface of trophozoites in both species, and longitudinal epicytical folds were only weakly developed. We also sequenced the small subunit rDNA from the gregarines collected from both hosts, which supported the establishment of two new Lankesteria species. Phylogenetic analyses of the new DNA sequences and those derived from other alveolates, demonstrated that both new species clustered in a strongly supported clade consisting of other Lankesteria species, Lecudina species, and some environmental sequences. These morphological and molecular phylogenetic data suggested that improved knowledge of gregarine diversity could lead to the recognition of more than one distinct clade (genus) of gregarines within ascidian hosts.     

Optimization of culture conditions and medium components for the production of mycelial biomass and exo-polysaccharides with Cordyceps militaris in liquid culture

Both crude exo-biopolymers and mycelial biomass, produced by liquid culture of Cordyceps species, are believed to possess several potential health benefits. As a result of its known biological activities, Cordyceps militaris has been extensively characterized in regards to potential medicinal applications. However, optimized liquid culture conditions for enhanced polysaccharide productivity have yet to be developed, which is a necessary step for industrial applications. Therefore, in this study, the liquid culture conditions were optimized for maximal production of mycelial biomass and exo-polysaccharide (EPS) by C. militaris. The effects of medium composition, environmental factors, and C/N ratio were investigated. Among these variables 80 g, glucose; 10 g, yeast extract; 0.5 g, MgSO₄·7H₂O; and 0.5 g, KH₂PO₄ in 1 L distilled water were found to be the most suitable carbon, nitrogen, and mineral sources, respectively. The optimal temperature, initial pH, agitation, and aeration were determined to be 24°C, uncontrolled pH, 200 rpm, and 1.5 vvm, respectively. Under these optimal conditions, mycelial growth in shake flask cultures and 5 L jar bioreactors was 29.43 and 40.60 g/L, respectively, and polysaccharide production in shake flask cultures and 5 L jar bioreactors was 2.53 and 6.74 g/L, respectively.     

The importance of gobies (Gobiidae,Teleostei) as hosts and transmitters of parasites in the SW Baltic

The parasite fauna of five goby species (Gobiidae, Teleostei) was investigated in the Baltic Sea during the period 1987 to 1990. 13 parasite species were found in samples from the Lübeck Bight:Bothriocephalus scorpii, Schistocephalus sp. (Cestoda);Cryptocotyle concavum, Cryptocotyle lingua, Podocotyle atomon, Derogenes varicus (Digenea);Hysterothylacium sp. (cf.auctum),Contracaecum sp.,Anisakis simplex (Nematoda);Corynosoma sp.,Echinorhynchus gadi, Neoechinorhynchus rutili, Pomphorhynchus laevis (Acanthocephala). The number of parasite species were: 10 in the sand gobyPomatoschistus minutus, 8 in the black gobyGobius niger, 7 in the two-spotted gobyGobiusculus flavescens, 6 in the common gobyPomatoschistus microps, and 5 in the painted gobyPomatoschistus pictus. Neoechinorhynchus rutili occurred only inP. minutus, andCorynosoma sp. only inG. niger. The extent to which the gobies were parasitized clearly depended on the respective ways of life and, moreover, on the kind of prey ingested by the hosts. Additionally, the age of the hosts might be important. The highest rate of parasitism, more than 60%, was reached byHysterothylacium sp. inG. niger and byCryptocotyle concavum inP. microps. Infestation incidence lay mostly below 40% which means a satellite species status (Holmes, 1991). The number of parasite species was highest in summer; the highest intensities of single parasites occurred in spring (Podocotyle atomon) or autumn (Crytocotyle concavum).Bothriocephalus scorpii, Hysterothylacium sp. andPodocotyle infested their juvenile hosts very early, but onlyHysterothylacium was accumulated byG. niger during its whole life span, whereasBothriocephalus persisted also in older gobies in low intensities. The cercariae ofCryptocotyle spp. penetrate actively into their hosts; all the other parasites named were transmitted in larval form by prey organisms which consisted mainly of planktonic and benthic crustaceans. The gobies were final hosts for only 5 parasites; but two species may be transmitted to larger fish, and 6 species to sea birds or mammals. The parasite community of the five gobies may possibly be taken to characterize the ecological quality of the environment of the Lübeck Bight.     

Nestmate discrimination and cuticular profiles of a temporary parasitic ant Lasius sp. and its host L. fuliginosus (Hymenoptera, Formicidae)

Workers of the temporary parasitic ant Lasius sp. were perfectly compatible with their hosts Lasius fuliginosus. Aggression was never seen between allospecific nestmates in either field or laboratory. In the laboratory, trophallaxis and allogrooming between allospecific nestmates were statistically more frequent than that between conspecifics. These ants were highly aggressive toward individuals of either species from another mixed colony located 1 km away, indicating that they discriminated nestmates and nonnestmates regardless of the species to which they belonged. No aggressive actions, however, were observed between two neighboring mixed nests located 5 m apart. We have not considered, however, the relationship of the two neighboring nests: the two nests may have been two subunits of the same family or sister colonies, or may have been different colonies but discriminated from distant aliens. Gas chromatography analysis on the total cuticular extracts showed that Lasius sp. and L. fuliginosus possess distinct cuticular profiles, even though they were collected from the same mixed colony. Received: December 13, 1999 / Accepted: April 25, 2000     

Diatom symbionts in larger foraminifera from Caribbean hosts

Larger foraminifera, as a group, are the hosts for many different types of algae. The majority of modern families host small (< 10 μm) pennate endosymbiotic diatoms. In these, the host/symbiont relationship is not finical. Studies from hosts harvested from Indo-Pacific habitats have shown some symbiont species are found with greater frequency than others, but any one of twenty different diatom species may be found in the same host. This study examined the endosymbionts from two diatom-bearing hosts from the Caribbean, Amphistigina gibbosa and Heterostegina antillarum. A high number of hosts harbored more than one symbiont species. A new variety of Nitzschia frustulum was involved in many (33%) of the symbioses. Many of the isolates of this taxon, if it is to be considered only one, are extremely aberrant forms for the genus. Another common (18%) isolate was an unidentified species of Navicula. A highly variable Amphora sp. was also isolated. These taxa will require more detailed examination in the future. Many of the isolates from the Caribbean hosts also were known as endosymbionts from previous studies: Nitzschia laevis N. frustulum var. symbiotica N. valdestriata, Amphora tenerrima, Navicula muscatini, Cocconeis andersoni and Fragillaria shiloi. A number of endosymbiotic species, common in Indo-Pacific hosts, were not found in the Caribbean hosts studied. The results of the present study are in consonance with those of previous studies.     

Acephaline Gregarines of Earthworms—Additions to the British Records

SYNOPSIS. 30 acephaline gregarines, including 4 new species, were identified and described from the perivisceral coelom and vesiculae seminales of 13 British earthworm species. The newly described monocystids were Apolocystis megagranulata n. sp. and Nematocystis dendrobaenae n. sp. from Dendrobaena rubida f. subrubicunda and D. mammalis, Monocystis mammaliae n. sp. from D. mammalis, and Zygocystis ictericae n. sp. from Allolobophora icterica f. typica. These 3 earthworm hosts have not been previously examined for monocystids in Britain.     

Permian bryophytes of Western Gondwanaland from the Paraná Basin in Brazil

Abstract: This study involves bryophyte fossils from the Teresina Formation (Permian–Guadalupian) collected in the Rio Preto Quarry in the state of Paraná, southern Brazil. Two new genera are proposed, with two new species: Capimirinus riopretensis sp. nov. and Yguajemanus yucapirus sp. nov. C. riopretensis sp. nov. included a lateral sporophyte that was organically attached to a gametophyte and is thus one of the oldest pleurocarpous fossils known today. This discovery provides clear evidence of the evolution of mosses during the Permian, because an acrocarpous moss from this period was also found in India. The preservation of such a fragile structure demonstrates that no prefossilization displacement took place and that the sediments accumulated in a low‐energy, fresh‐water environment, similar to a lake fed by rain, and isolated from the direct influence of the sea. Campimirinus riopretensis may be placed within the earlier pleurocarps, and Yguajemanus yucapirus sp. nov. may belong to the Bryalean families. However, we were not able to place neither of these species in any existing family today.