Widespread mermithid nematode parasitism of Cretaceous insects.

Mermithid nematodes are obligate invertebrate parasites dating back to the Early Cretaceous. Their fossil record is sparse, especially before the Cenozoic, thus little is known about their early host associations. This study reports 16 new mermithids associated with their insect hosts from mid-Cretaceous Kachin amber, 12 of which include previously unknown hosts. These fossils indicate that mermithid parasitism of invertebrates was already widespread and played an important role in the mid-Cretaceous terrestrial ecosystem. Remarkably, three hosts (bristletails, barklice, and perforissid planthoppers) were previously unknown to be parasitized by mermithids both past and present. Furthermore, our study shows that in contrast to their Cenozoic counterparts, Cretaceous nematodes including mermithids are more abundant in non-holometabolous insects. This result suggests that nematodes had not completely exploited the dominant Holometabola as their hosts until the Cenozoic. This study reveals what appears to be a vanished history of nematodes that parasitized Cretaceous insects.

A mid-Cretaceous ambrosia fungus, Paleoambrosia entomophila gen. nov. et sp. nov. (Ascomycota: Ophiostomatales) in Burmese (Myanmar) amber, and evidence for a femoral mycangium.

An ambrosia fungus is described from filamentous sporodochia adjacent to a wood-boring ambrosia beetle (Coleoptera: Curculionidae: Platypodinae) in mid-Cretaceous Burmese amber. Yeast-like propagules and hyphal fragments of Paleoambrosia entomophila gen. nov. et sp. nov. occur in glandular sac mycangia located inside the femur of the beetle. This is the first record of a fossil ambrosia fungus, showing that symbiotic associations between wood-boring insects and ectosymbiotic fungi date back some 100 million years ago. The present finding moves the origin of fungus-growing by insects from the Oligocene to the mid-Cretaceous and suggests a Gondwanan origin.

Hexamermis popilliae n. sp. (Nematoda: Mermithidae) parasitizing the Japanese beetle Popillia japonica Newman (Coleoptera: Scarabaeidae) in Italy.

A new species of mermithid nematode, Hexamermis popilliae n. sp. (Nematoda: Mermithidae) is described from the Japanese beetle Popillia japonica Newman in Italy, an area of new introduction for this invasive pest. The combination of the following characters separates H. popilliae from other members of the genus Hexamermis Steiner, 1924: adult head obtuse; amphidial pouches slightly posterior to lateral head papillae in female but adjacent to lateral head papillae in males; amphidial openings large, well developed; amphidial pouches elliptical in females and oblong in males; cuticular vulvar cone well developed, vulvar lips greatly reduced or lacking, vagina curved at tip where meeting uteri, without reverse bend (not S-shaped), spicules slightly curved, with a slight bend in the basal portion, approximately equal to body width at cloaca. This is the first record of a species of Hexamermis parasitizing the Japanese beetle Popillia japonica. The only previous mention of mermithid nematodes from P. japonica was an undescribed species of Psammomermis in North America. Hexamermis popilliae will be evaluated as a potential biological control agent in an integrated control program of the Japanese beetle in Italy.

An asterid flower from neotropical mid-Tertiary amber.

Fossils preserved in amber may provide significant palaeoevolutionary and biogeographical data regarding the evolution of life on Earth(1). Although amber is particularly noted for its detailed preservation of arthropods, the same degree of preservation can be found for vascular plant remains(2). Mid-Tertiary Dominican amber is a rich source for such fossils, and representatives of several angiosperm families have been described. However, no fossilized examples of the large asterid plant clade have yet been reported. Here we describe the first fossil neotropical flowers found in amber from a representative of the asterids. The asterids are one of the largest lineages of flowering plants, containing groups such as the sunflower, potato, coffee and mint families, totalling over 80,000 species(3). The new fossils are only known as flowers, more precisely corollas with stamens and styles. We here describe them as a new species, Strychnos electri sp. nov, in the plant family Loganiaceae (Gentianales).

The Geological Record of Parasitic Nematode Evolution.

This chapter discusses the evolutionary history of nematode parasites of invertebrates, vertebrates and plants based on fossil remains in amber, stone and coprolites dating from the Palaeozoic to the Holocene. The earliest parasitic nematode is a primitive plant parasite from the Devonian. Fossil invertebrate-parasitic nematodes first appeared in the Early Cretaceous, while the earliest fossil vertebrate-parasitic nematodes are from Upper Triassic coprolites. Specific examples of fossil nematode parasites over time are presented, along with views on the origin and evolution of nematodes and their hosts.

Can parasites halt the invader? Mermithid nematodes parasitizing the yellow-legged Asian hornet in France.

Since its introduction in France 10 years ago, the yellow-legged Asian bee-hawking hornet Vespa velutina has rapidly spread to neighboring countries (Spain, Portugal, Belgium, Italy, and Germany), becoming a new threat to beekeeping activities. While introduced species often leave behind natural enemies from their original home, which benefits them in their new environment, they can also suffer local recruitment of natural enemies. Three mermithid parasitic subadults were obtained from V. velutina adults in 2012, from two French localities. However, these were the only parasitic nematodes reported up to now in Europe, in spite of the huge numbers of nests destroyed each year and the recent examination of 33,000 adult hornets. This suggests that the infection of V. velutina by these nematodes is exceptional. Morphological criteria assigned the specimens to the genus Pheromermis and molecular data (18S sequences) to the Mermithidae, due to the lack of Pheromermis spp. sequences in GenBank. The species is probably Pheromermis vesparum, a parasite of social wasps in Europe. This nematode is the second native enemy of Vespa velutina recorded in France, after a conopid fly whose larvae develop as internal parasitoids of adult wasps and bumblebees. In this paper, we provide arguments for the local origin of the nematode parasite and its limited impact on hornet colony survival. We also clarify why these parasites (mermithids and conopids) most likely could not hamper the hornet invasion nor be used in biological control programs against this invasive species.

Monsters of the sea serpent: parasites of an oarfish, Regalecus russellii.

Examination of a small portion of the viscera of an oarfish ( Regalecus russellii ) recovered from Santa Catalina Island, southern California, revealed numerous tetraphyllidean tapeworm plerocercoids, Clistobothrium cf. montaukensis; 2 juvenile nematodes, Contracaecum sp.; and a fragment of an adult acanthocephalan, family Arhythmacanthidae. This suggests that the fish was relatively heavily parasitized. The presence of larval and juvenile worms suggests that oarfish are preyed upon by deep-swimming predators such as the shortfin mako shark, Isurus oxyrinchus , known to be a definitive host for the adult tapeworm, and also by diving mammals such as sperm whales, Physeter catodon L., hosts of Contracaecum spp. nematodes.

Parasitylenchus bifurcatus n. sp. (Tylenchida: Allantonematidae) parasitizing Harmonia axyridis (Coleoptera: Coccinellidae).

BACKGROUND: The harlequin ladybird, Harmonia axyridis Pallas (Coleoptera: Coccinellidae) is native to central and eastern Asia and was purposely introduced into Europe to control aphids. While it proved to be a good biological control agent, its rapid spread and buildup of large populations made it a nuisance, since it overwinters in homes, emits unpleasant odors, stains fabrics, occasionally bites humans and feeds on apples, pears and grapes. Aside from the above, the ravenous appetite of H. axyridis results in their consumption of harmless native insects, including even other ladybird beetles. A study of the natural enemies of H. axyridis in Denmark revealed the presence of nematodes. The present study describes this nematode parasite and discusses aspects of its development and ecology. METHODS: Adult harlequin ladybird beetles were collected from March to November from four localities in Copenhagen on different plant species. In addition, groups of last-instar larvae and pupae (n = 50) were examined for the presence of nematodes. Living and recently dead nematodes were removed from adult H. axyridis in 0.5% saline solution, the nematodes were then heat killed (at 75C), fixed in 5% formalin and transferred to glycerin on slides for further examination and measurements. RESULTS: A new species of Allantonematidae (Tylenchida), Parasitylenchus bifurcatus n. sp., is described from adults of the harlequin ladybird, Harmonia axyridis in Denmark. The new species is characterized by a straight stylet lacking basal thickenings, a bursa and a forked tail tip in the vermiform (infective) females and juvenile males. The new species is compared with P. coccinellinae previously described from ladybird beetles in France. Parasitism resulted in depletion of the fat body and partial or complete atrophy of the reproductive organs of the beetles. Infections occurred throughout the year with rates of parasitism reaching up to 35%. The rate increased to 60% when field-collected ladybirds were incubated for 30 days in the laboratory. CONCLUSIONS: The production of subsequent generations within the host with only the fertilized females (not the males) leaving the hosts and the absence of parasitism of the larvae and pupae is an impressive developmental modification of P. bifurcatus. It is proposed that the vermiform (infective) females pass from one adult host to another when the beetles are hibernating or in assemblage groups. Rates of parasitism show that P. bifurcatus could be a significant biological control agent of H. axyridis.

Palaeontology: the 165-million-year itch.

New flea-like fossils from China provide a rare, tantalizing glimpse of bizarre insects in the Cretaceous and Jurassic. Possibly the oldest flea-like animals known, they provide a challenge to the functional morphologist to infer which animals they may have targeted.

Halophilanema prolata n. gen., n. sp. (Nematoda: Allantonematidae), a parasite of the intertidal bug, Saldula laticollis (Reuter)(Hemiptera: Saldidae) on the Oregon coast.

BACKGROUND: It is rare to find terrestrial nematode lineages parasitizing arthropods inhabiting the intertidal or littoral zone of the oceans. During an ecological study along the Oregon dunes, an allantonematid nematode (Tylenchomorpha: Allantonematidae) was discovered parasitizing the intertidal shore bug, Saldula laticollis (Reuter)(Hemiptera: Saldidae). This shore bug is adapted to an intertidal environment and can survive short periods of submergence during high tides. The present study describes the nematode parasite and discusses aspects of its development, ecology and evolution. METHODS: Adults and last instar nymphs of S. laticollis (Hemiptera: Saldidae) were collected from the high intertidal zone among clumps of Juncus L. (Juncaceae) plants at Waldport, Oregon on October 3, 2011. The bugs were dissected in 1% saline solution and the nematodes killed in 1% Ringers solution and immediately fixed in 5% formalin (at 20°C). Third stage juveniles removed from infected hosts were maintained in 1% saline solution until they matured to the adult stage, molted and mated. RESULTS: Halophilanema prolata n. gen., n. sp. (Nematoda: Allantonematidae) is described from last instar nymphs and adults of the intertidal bug, Saldula laticollis on the Oregon coast. The new genus can be distinguished from other genera in the Allantonematidae by a stylet lacking basal knobs in both sexes, an excretory pore located behind the nerve ring, ribbed spicules, a gubernaculum, the absence of a bursa and the elongate-tubular shape of the ovoviviparous parasitic females. Studies of the organogenesis of Halophilanema showed development to third stage juveniles in the uterus of parasitic females. Maturation to the free-living adults and mating occurred in the environment. The incidence of infection of S. laticollis ranged from 0% to 85% depending on the microhabitat in the intertidal zone. CONCLUSIONS: Based on the habitat and morphological characters, it is proposed that Halophilanema adapted a parasitic existence fairly recently, evolutionarily speaking. It was probably a free-living intertidal or shore nematode that fed on microorganisms, especially fungi, in the intertidal habitat and became parasitic after saldids entered the environment. Halophilanema represents the first described nematode parasite of an intertidal insect.

Vetufebrus ovatus n. gen., n. sp. (Haemospororida: Plasmodiidae) vectored by a streblid bat fly (Diptera: Streblidae) in Dominican amber.

BACKGROUND: Both sexes of bat flies in the families Nycteribiidae and Streblidae (Diptera: Hippoboscoidea) reside in the hair or on the wing membranes of bats and feed on blood. Members of the Nycteribiidae transmit bat malaria globally however extant streblids have never been implemented as vectors of bat malaria. The present study shows that during the Tertiary, streblids also were vectors of bat malaria. RESULTS: A new haemospororidan, Vetufebrus ovatus, n. gen., n. sp., (Haemospororida: Plasmodiidae) is described from two oocysts attached to the midgut wall and sporozoites in salivary glands and ducts of a fossil bat fly (Diptera: Streblidae) in Dominican amber. The new genus is characterized by ovoid oocysts, short, stubby sporozoites with rounded ends and its occurrence in a fossil streblid. This is the first haemosporidian reported from a streblid bat fly and shows that representatives of the Hippoboscoidea were vectoring bat malaria in the New World by the mid-Tertiary. CONCLUSIONS: This report is the first evidence of an extant or extinct streblid bat fly transmitting malaria. Discovering a mid-tertiary malarial parasite in a fossil streblid that closely resembles members of a malarial genus found in nycteribiid bat flies today shows how little we know about the vector associations of streblids. While no malaria parasites have been found in extant streblids, they probably occur and it is possible that streblids were the earliest lineage of flies that transmitted bat malaria to Chiroptera.

Description of an early Cretaceous termite (Isoptera: Kalotermitidae) and its associated intestinal protozoa, with comments on their co-evolution.

BACKGROUND: The remarkable mutualistic associations between termites and protists are in large part responsible for the evolutionary success of these eusocial insects. It is unknown when this symbiosis was first established, but the present study shows that fossil termite protists existed in the Mesozoic. RESULTS: A new species of termite (Kalotermes burmensis n. sp.) in Early Cretaceous Burmese amber had part of its abdomen damaged, thus exposing trophic stages and cysts of diverse protists. Some protists were still attached to the gut intima while others were in the amber matrix adjacent to the damaged portion. Ten new fossil flagellate species in the Trichomonada, Hypermastigida and Oxymonadea are described in nine new genera assigned to 6 extant families. Systematic placement and names of the fossil flagellates are based on morphological similarities with extant genera associated with lower termites. The following new flagellate taxa are established: Foainites icelus n. gen. n. sp., Spiromastigites acanthodes n. gen. n. sp., Trichonymphites henis n. gen., n. sp., Teranymphites rhabdotis n. gen. n. sp., Oxymonas protus n. sp., Oxymonites gerus n. gen., n. sp., Microrhopalodites polynucleatis n. gen., n. sp., Sauromonites katatonis n. gen., n. sp., Dinenymphites spiris n. gen., n. sp., Pyrsonymphites cordylinis n. gen., n. sp. A new genus of fossil amoeba is also described as Endamoebites proterus n. gen., n. sp. Fourteen additional trophic and encystid protist stages are figured and briefly characterized. CONCLUSION: This represents the earliest fossil record of mutualism between microorganisms and animals and the first descriptions of protists from a fossil termite. Discovering the same orders, families and possibly genera of protists that occur today in Early Cretaceous kalotermitids shows considerable behaviour and morphological stability of both host and protists. The possible significance of protist cysts associated with the fossil termite is discussed in regards the possibility that coprophagy, as well as proctodeal trophallaxis, was a method by which some termite protozoa were transferred intrastadially and intergenerationally at this time.

The oldest fossil evidence of animal parasitism by fungi supports a Cretaceous diversification of fungal-arthropod symbioses.

Paleoophiocordyceps coccophagus, a fungal parasite of a scale insect from the Early Cretaceous (Upper Albian), is reported and described here. This fossil not only provides the oldest fossil evidence of animal parasitism by fungi but also contains morphological features similar to asexual states of Hirsutella and Hymenostilbe of the extant genus Ophiocordyceps (Ophiocordycipitaceae, Hypocreales, Sordariomycetes, Pezizomycotina, Ascomycota). Because species of Hypocreales collectively exhibit a broad range of nutritional modes and symbioses involving plants, animals and other fungi, we conducted ancestral host reconstruction coupled with phylogenetic dating analyses calibrated with P.coccophagus. These results support a plant-based ancestral nutritional mode for Hypocreales, which then diversified ecologically through a dynamic process of intra- and interkingdom host shifts involving fungal, higher plant and animal hosts. This is especially evident in the families Cordycipitaceae, Clavicipitaceae and Ophiocordycipitaceae, which are characterized by a high occurrence of insect pathogens. The ancestral ecologies of Clavicipitaceae and Ophiocordycipitaceae are inferred to be animal pathogens, a trait inherited from a common ancestor, whereas the ancestral host affiliation of Cordycipitaceae was not resolved. Phylogenetic dating supports both a Jurassic origin of fungal-animal symbioses within Hypocreales and parallel diversification of all three insect pathogenic families during the Cretaceous, concurrent with the diversification of insects and angiosperms.

A new species of Phyllopsora (Lecanorales, lichen-forming Ascomycota) from Dominican amber, with remarks on the fossil history of lichens.

Phyllopsora dominicanus sp. nov. (Bacidiaceae, Lecanorales, lichen-forming Ascomycota) is described and illustrated from Dominican amber. The diagnostic features of the lichen include a minute subfolious thallus of lacinulate, ascending squamules, a well-developed upper cortex, and a net-like pseudocortex on the lower surface. The algal symbionts are unicellular green algae, forming a distinct layer immediately below the upper cortex. The fossil demonstrates that distinguishing features of Phyllopsora have remained unchanged for tens of millions of years. The fossil also provides the first detailed views of mycobiont-photobiont contacts in Tertiary green algal lichens. The mycobiont hyphae formed apical and intercalary appressoria by pressing closely against the photobiont cells. This indicates that a conserved maintenance of structure is also seen in the fine details of the fungal-algal interface.

Allomermis solenopsi n. sp. (Nematoda: Mermithidae) parasitising the fire ant Solenopsis invicta Buren (Hymenoptera: Formicidae) in Argentina.

Allomermis solenopsi n. sp. (Mermithidae: Nematoda) is described from the fire ant Solenopsis invicta Buren (Hymenoptera: Formicidae) in Argentina. Diagnostic characters of the new species include stiff and erect processes on the surface of the mature egg, small female amphids, extension of the latero-medial rows of male genital papillae beyond the middle rows, an obliquely truncate spicule tip and a ventrally swollen male terminus. This is the first record of Allomermis Steiner, 1924 from South America and the first host record for members of this genus. Previous records of mermithids from Solenopsis spp. are summarised. The placement in Allomermis was confirmed by molecular analyses based on nuclear 18S ribosomal DNA sequences, the first such molecular framework for the Mermithidae. The possible life-cycle of the parasite is discussed, with the aim of using A. solenopsi as a biological control agent for fire ants in the United States.

Evidence of mycoparasitism and hypermycoparasitism in Early Cretaceous amber.

Evidence of mycoparasitism and hypermycoparasitism is demonstrated in Early Cretaceous Burmese amber. The agaric, Palaeoagaracites antiquus gen. sp. nov., is parasitized by the mycoparasite, Mycetophagites atrebora gen. sp. nov., which in turn is parasitized by the hyperparasite, Entropezites patricii gen. sp. nov. This discovery shows that sophisticated patterns of fungal parasitism were well developed some 100 Myrago.

A non-gilled hymenomycete in Cretaceous amber.

Palaeoclavaria burmitis gen. et sp. nov. (Palaeoclavariaceae fam. nov, Hymenomycetes) is described from a series of fruit bodies and hyphae in Cretaceous amber from Burma (about 100 Myr). This is the first fossil record of the Aphyllophorales and establishes certain basic morphological and ecological characters for the group.

First record of a Mermithidae (Nematoda) from the meloid beetle Meloe violaceus Marsham, 1802 (Coleoptera: Meloidae).

A new record of nematode parasitism of meloid beetles is reported and all earlier records are summarised. Rates of parasitism could be influenced by the toxic compound cantharidin that these beetles possess.

Amber: the organic gemstone.

Resins are produced by woody plants on a worldwide basis. We have found several distinct classes of modern diterpenoid resins based phenomenologically on the solid-state (13)C NMR spectra of the bulk material. Resin fossilizes over millions of years into a robust material sometimes called amber. We have characterized several hundred samples of fossil resin by solid-state (13)C NMR spectroscopy. We can relate one globe-spanning group of fossil resins to the modern genus Agathis, based on spectral evolution over time. A second large group has not been related with certainty to specific modern plants. Fossil resins from Europe fall into two categories, the famous Baltic ambers and another that resembles the Agathis group. Fossil resins from the Americas and Africa are closely related to the modern genus Hymenaea. Based on spectral distinctions, fossil resin found in an archaeological context sometimes can be assigned to a specific geographical origin on the basis of its (13)C NMR spectrum.