Porcine reproductive and respiratory syndrome virus (PRRSV) infection spreads by cell-to-cell transfer in cultured MARC-145 cells, is dependent on an intact cytoskeleton, and is suppressed by drug-targeting of cell permissiveness to virus infection

Background

Porcine reproductive and respiratory syndrome virus (PRRSV) is the etiologic agent of PRRS, causing widespread chronic infections which are largely uncontrolled by currently available vaccines or other antiviral measures. Cultured monkey kidney (MARC-145) cells provide an important tool for the study of PRRSV replication. For the present study, flow cytometric and fluorescence antibody (FA) analyses of PRRSV infection of cultured MARC-145 cells were carried out in experiments designed to clarify viral dynamics and the mechanism of viral spread. The roles of viral permissiveness and the cytoskeleton in PRRSV infection and transmission were examined in conjunction with antiviral and cytotoxic drugs.

Results

Flow cytometric and FA analyses of PRRSV antigen expression revealed distinct primary and secondary phases of MARC-145 cell infection. PRRSV antigen was randomly expressed in a few percent of cells during the primary phase of infection (up to about 20–22 h p.i.), but the logarithmic infection phase (days 2–3 p.i.), was characterized by secondary spread to clusters of infected cells. The formation of secondary clusters of PRRSV-infected cells preceded the development of CPE in MARC-145 cells, and both primary and secondary PRRSV infection were inhibited by colchicine and cytochalasin D, demonstrating a critical role of the cytoskeleton in viral permissiveness as well as cell-to-cell transmission from a subpopulation of cells permissive for free virus to secondary targets. Cellular expression of actin also appeared to correlate with PRRSV resistance, suggesting a second role of the actin cytoskeleton as a potential barrier to cell-to-cell transmission. PRRSV infection and cell-to-cell transmission were efficiently suppressed by interferon-γ (IFN-γ), as well as the more-potent experimental antiviral agent AK-2.

Conclusion

The results demonstrate two distinct mechanisms of PRRSV infection: primary infection of a relatively small subpopulation of innately PRRSV-permissive cells, and secondary cell-to-cell transmission to contiguous cells which appear non-permissive to free virus. The results also indicate that an intact cytoskeleton is critical for PRRSV infection, and that viral permissiveness is a highly efficient drug target to control PRRSV infection. The data from this experimental system have important implications for the mechanisms of PRRSV persistence and pathology, as well as for a better understanding of arterivirus regulation.     

A carboniferous chondrichthyan assemblage from residues within a Triassic karst system at Cromhall quarry,Gloucestershire, England

Abstract. Sixteen different Lower Carboniferous (Tournaisian Courceyan to Chadian age, Mississippian) chondrichthyan teeth types have been extracted from Triassic erosional/aeolian fills in shallow karst systems found in the limestone quarry at Cromhall, Gloucestershire, England. These Carboniferous teeth have been found within a much larger assemblage of disarticulated bones and teeth belonging to small late Triassic terrestrial reptiles, for which Cromhall quarry is famous. The Carboniferous teeth are derived fossils, released during subaerial dissolution of the surrounding limestones. Owing to low specimen numbers and uncertainty of intraspecific character, 10 types of teeth are left in open nomenclature, although it is likely they represent new taxa. They belong to Jalodus, Ctenacanthiformes, Protacrodus, Orodus, Chomatodus, Petalodontidae, Euchondrocephali?, Helodus and Psephodus. A new genus and species, Cromhallia parvunda, of indeterminate phylogenetic relationships, is also erected. The assemblage includes also an identified suite comprising Thrinacodus ferox, Bransonella cf. nebraskensis and Stethacanthus cf. altonensis. The identification of all fish teeth found in the Cromhall assemblages as derived fossils from the Carboniferous removes any ambiguity regarding the fully terrestrial nature of the late Triassic fauna preserved in the residue‐bearing karst systems.     

Long‐term impacts of invasive species on a native top predator in a large lake system

1. Declining abundances of forage fish and the introduction and establishment of non‐indigenous species have the potential to substantially alter resource and habitat exploitation by top predators in large lakes. 2. We measured stable isotopes of carbon (δ¹³C) and nitrogen (δ¹⁵N) in field‐collected and archived samples of Lake Ontario lake trout (Salvelinus namaycush) and five species of prey fish and compared current trophic relationships of this top predator with historical samples. 3. Relationships between δ¹⁵N and lake trout age were temporally consistent throughout Lake Ontario and confirmed the role of lake trout as a top predator in this food web. However, δ¹³C values for age classes of lake trout collected in 2008 ranged from 1.0 to 3.9‰ higher than those reported for the population sampled in 1992. 4. Isotope mixing models predicted that these changes in resource assimilation were owing to the replacement of rainbow smelt (Osmerus mordax) by round goby (Neogobius melanostomus) in lake trout diet and increased reliance on carbon resources derived from nearshore production. This contrasts with the historical situation in Lake Ontario where δ¹³C values of the lake trout population were dominated by a reliance on offshore carbon production. 5. These results indicate a reduced capacity of the Lake Ontario offshore food web to support the energetic requirements of lake trout and that this top predator has become increasingly reliant on prey resources that are derived from nearshore carbon pathways.     

Cheilostomatous Bryozoa from Vanuatu

The present account of some Bryozoa from Vanuatu is the first study of reef-flat species from the eastern Coral Sea. Based on samples collected from several shallow water localities on the island of Efate, a total of 92 species is described, including a new family, three new genera and 20 new species. Some of the newly recognized species are the result of taxonomic revision stimulated by these new samples, while others are entirely new to science. Of the new taxa described here, 16 are presently known only from Vanuatu. The total number of species recorded does not accurately represent the true taxonomic diversity of the Cheilostomatida of Vanuatu, and further sampling, at other localities and in other habitats, is needed before it can be reasonably estimated. The fauna is predominantly Indo-West Pacific in character with a high proportion of species apparently known only in the Southwest Pacific.     

Anamorphosis in millipedes (Diplopoda)—the present state of knowledge with some developmental and phylogenetic considerations

A review of the postembryonic development of millipedes (Diplopoda) is given, based mainly on published information. Original observations are, however, also included. Millipedes hatching from the pupoid usually have three pairs of legs; during their postembryonic growth they acquire more segments and more legs. This process is known as anamorphosis. Three types of anamorphosis are recognized. In euanamorphosis, every moult is accompanied by addition of new segments, even after the attainment of sexual maturity. In hemianamorphosis, the addition of new segments goes on until a certain stadium, and further moults take place without addition of segments. In teloanamorphosis, the addition of segments stops at a certain stadium (the adult, and ultimate, stadium) after which no further moults occur. Available information on anamorphosis in each of the millipede orders is reviewed. General patterns are emphasized, but variations are also considered in detail. It is shown that the so-called ‘law of anamorphosis’ is valid only for the ‘ring-forming’ millipedes (Merocheta and Juliformia) in which tergites, pleurites, and sternites of each diplosegment are firmly fused into a complete ‘ring’, and for some other forms (Polyzoniida, Chordeumatida), where there is a constant relationship between rings and legs. The chapter on the order Julida is particularly detailed and includes discussions of patterns in the variation and a section on periodomorphosis. The general chapter on developmental patterns includes inter alia an interpretation of the variations in millipede anamorphosis in terms of the ‘biometabolic modi’ of Remane. The hypothetical ancestral millipede is shown to have developed by hemianamorphosis. Euanamorphosis was acquired by the ancestral species of Helminthomorpha. Within this clade, Chordeumatida and Merocheta have secondarily become teloanamorphic, whereas some Juliformia seem to have returned to hemianamorphosis. The contrasting principles of elongation and contraction, subject of much debate among diplopodologists, are shown both to have played a role in the course of millipede evolution.