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Welcome to the Newschoolers forums! You may read the forums as a guest, however you must be a registered member to post. Register to become a member today! A quick word on blocking ads. It looks like you are using an ad blocker. That's okay. Who doesn't? But without advertising revenue, we can't keep making this site awesome. Click the link below for instructions on disabling adblock. How to disable ad-blocker for Newschoolers. I don't care about Newschoolers. I just want free content and no ads! Register Lost password? Move to Category. Close Save. Member of the Month BallClapper September, Jul 28 AM. Dain bramaged. Little known fact about super pipes: The super pipe gets its dimentions from king kongs peinis. The first ever superpipe was in down town New York City and it was make by none other than King Kong himself. You see, when the helicopters finaly made him fall off the Empire State Building he went crashing to the ground. And of course when somthing as huge as King Kong falls a few thousand feet it leaves a dent in the ground. Well who would'a known that King Kong's schlong would have left a perfect, yard long, 18 foot deep ditch in the ground? Turns out the day after the city workers removed the body, Manhattan was engulfed in a massive blizzard. The penial imprint was filled with snow. The local kids were quick to recognize this great natural feature and went to town skiing and boarding on it. The mountain people didn't like this at all so they sent their cronnies out with messuring sticks and tapes, made some carful observatios and took some notes. The fruits of their research was the super pipe dragon. Everyone seems to know that last part, but I gotta give it up for my man Kong and his dong, because without either of them we'd still be riding pussy little halfpipes. Jul 28 PM. Oo LoL that was Funny King Kong's shlong How does it feel to ski where someone else's member has been. If a man is standing in the middle of the forest speaking and there is no woman around to hear him - Is he still wrong? Darryl Hunt aka - highschool representing the H. Rummor has it that mammoth will have 2 super pipes next season the main park super pipe will be replaced with one of the 22' monsters. The super pipe was invented by Al Clark, whistler local, sweet snowboarder. Not sure exactly but I think the walls are 16' and bigger to be a super pipe. I thought Al Gore invented the superpipe. Long before Hood got dragons all the pipes were hand shaped by crews of 15 diggers. The cats would come in once at the begining of each session rough out the pipe in the glacier then it would take 2 full days by hand to shape it. The last great year of monster hand dug pipes was 97 when Hood got a regular sized dragon. Al's 'superpipe' camp didnt come along until 98 or Jul 29 AM. And they said that the walls are built a few degrees below vert although I'm not exactly sure why. Jul 30 PM. Jul 31 AM. The first pipe dragon was made by, shit, i cant remember his name, i need to check my history on this one. Anyway, it was made by a guy in Breckenridge, and he used it to cut what is recognized as the first superpipe with walls 15 feet 6' high. The transition was based on a 15 foot vert ramp used for a professional skateboard competition. The dimensions now have been altered to be better for skis and snowboards to handle. Aug 1 AM. With all due respect to the game, I'm the P-H-enom Not ready for prime time, be-yond, extinction Change your way of thinkin, or be-gone fast the fuck out, somethin stinkin Could it be the skunk, or could it be that body in the trunk of my Lincoln, Continental style pop the pussy like a pimple I'm fed up, I put it in your ear and fuck ya head up Turnin up the temperature, hold them kids that entered the 36th, master mix shit Biohazardous, pretentious Do it for the chemically imbalanced State your business, pay me at the door Iron Man, hear me roar on twelve inches Shell shocked soldier in the trenches Fire in the hole game commences Third string rappers play the benches Reload, there'll be no repentance for souls Just life sentence, with no chance for parole and that's real. Ill flow. Above someone said the inventer was from Breck and it was 15 ft pipe. His name is Doug and is from Berthud Co. His first machine waas only 8ft modeled after vert ramps at the time He didnt build a superdragon until summer of That machine was modeled after the large hand dug pipes of the early to mid 90's at the various Mt hood summer camps. That machine was first used at Hood then Whistler. Whistler I believe bought the prototype, Mammoth bought the first one in production and Breck bought the second. Aug 1 PM. The king kong shit is still more believable then this Al gore and Windells mombojumbo. Canadian think of everything and then Americans just take it from us. And ya know that big long penis shaped arm which is on all those space crafts in space was created by canadians. I dont think ppl realize how big sunday rivers pipe is until they actually see it Al Clark was the guy who founded the superpipe idea. There may have been some in the past that just happened, but he was the guy with the idea. Also, a superpipe doesn't have 6 ft of vert, what the fuck? It usually just goes to vert or is just under vert, usually just under vert, thats the way pipes work best. You could'nt even ski in a pipe with 6 ft of vert. All times are Eastern

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Official websites use. Share sensitive information only on official, secure websites. E-mail: gilda. The enigmatic origins and transmission events of the gibbon ape leukemia virus GALV and its close relative the koala retrovirus KoRV have been a source of enduring debate. Bats and rodents are each proposed as major reservoirs of interspecies transmission, with ongoing efforts to identify additional animal hosts of GALV-KoRV-related retroviruses. Included among these hosts are two African rodents, revealing the first appearance of this clade beyond the Australian and Southeast Asian region. Keywords: cross-species virus transmission, Australian animal health, GALV gibbon-ape leukemia virus, KoRV koala retrovirus, phylogenetic analysis of endogenous retroviruses, NGS high-throughput sequencing data mining virus discovery. The territories of koalas and gibbons do not overlap and are separated by a biogeographical faunal boundary across a large body of water, preventing direct viral transmission between these hosts Brockelman and Geissmann First, GALV-KoRV-related retroviruses were discovered in taxa other than koalas and gibbons in recent years, initially in an Australian rodent with subspecies in Indonesia Simmons et al. Second, rodents and bats together comprise almost half of all mammalian species, and phylogenomic analyses have revealed that both bats and rodents are significantly involved in the history of retrovirus transmission between other mammalian species Hayward et al. In particular, the evolutionary distance between KoRV, GALV, and other viral relatives is large enough that there are almost certainly other virus—host associations belonging to this clade that remain undiscovered. When considering the potential for zoonotic transmission and pathogenicity in humans or other animals of domestic, economic, and ecological importance, questions regarding the origin, transmission, and hosts of GALV-KoRV-related retroviruses need to be addressed. KoRV is widespread in koala populations, particularly in northeastern Australia Quigley et al. Importantly, that virus, the Hervey pteropid gammaretrovirus HPG , was previously shown to be capable of infecting human cells in vitro Hayward et al. This is a result of the hallmark of retrovirus replication where the retroviral proviral DNA precursor is inserted into the genome of the host Johnson When this happens in germline cells that become new offspring, the retrovirus becomes an endogenized, heritable genetic element. Over the course of evolutionary history, vertebrate genomes have become littered with the remains of past retroviral infections Johnson Endogenous retroviruses are subject to genetic drift and tend to become defective over many host generations Johnson KoRV subtype A KoRV-A is a recently integrated endogenous retrovirus in the koala gene pool, and still generates infectious viral particles that can be transmitted between animals Tarlinton et al. This receptor is ubiquitously expressed on mammalian cells, and a permissivity motif within its sequence can be used to infer potential susceptibility to infection by GALV-KoRV-related retroviruses Schneiderman et al. In this study, we aimed to leverage the vast amount of publicly available data in nucleotide sequence read archives SRA and genome assemblies to identify unreported GALV-KoRV-related retroviruses and their hosts. Novel retroviral sequences are phylogenetically analysed to infer their evolutionary relatedness to known gammaretroviruses, and the PiT-1 receptor sequences of their putative hosts are evaluated to infer their potential susceptibility to infection by GALV-KoRV-related retroviruses. For many species, no publicly available SRA records exist. Hits representing novel retroviral sequences were from the rodents Mastacomys fuscus Broad-toothed rat , Pseudomys apodemoides Silky mouse , P. These rodents occupy diverse regions across the Australian continent that collectively include all Australian states and the Northern Territory Fig. Some contigs from individual species were found to overlap. The natural ranges of these rodents are indicated by the brown-shaded region overlayed on the map of Africa or Australia Aplin and Woinarski , Burbidge and Woinarski , Cassola , Cassola and Menkhorst , Granjon , Burbidge a , b , Woinarski and Burbidge a , b. The approximate locations of the collected samples are indicated by the dots. Kelly et al. Mammalian species samples in which novel GALV-KoRV-related retroviruses were identified using the Hervey pteropid gammaretrovirus receptor binding domain nucleotide sequence. Overlapping contigs from individual species were included in the same phylogenies, while contigs that did not overlap were analysed separately. Phylogenies for gag, pol , and env retroviral sequences from each rodent species are included in SI Figs1—4. Additional phylogenies for several of these species are included in SI Fig 5—7. This indicates that these contigs were derived from defective endogenous retroviruses. Only one species, P. Maximum likelihood phylogenies of regions of nucleotide sequences from the env genes of a Mastacomys fuscus ; b Zyzomys argurus ; c Pseudomys apodemoides ; d Pseudomys bolami ; e Pseudomys delicatulus ; and f Pseudomys johnsoni. Silhouettes represent the animal hosts rodents, koalas, primates, fruit bats, and microbats. All branches are scaled according to the number of nucleotide substitutions per site as indicated by the scale bars. Bootstrap support values are shown at the nodes. The number of nucleotide positions in the multiple sequence alignments used to generate phylogenies a—f are , , , , , and respectively. All Australian rodent datasets contain multiple contigs from one or more of the core retroviral genes gag, pol , and env that phylogenetically clustered within the GALV-KoRV-related retrovirus clade. No contigs were identified which were closely and immediately basal to the KoRV sub-clade. For env sequences Fig. For gag and pol , contigs clustered at variable positions within the phylogeny, including in some cases, at a position basal to McERV SI Figs 1—7. Together, these contigs may represent multiple divergent gammaretroviruses, or individual recombinant retroviruses that include one or two genes derived from a GALV-KoRV-related retrovirus, and the remainder from a more distantly related gammaretrovirus. We extended our search for unreported GALV-KoRV-related retroviruses to all parts of the world, searching within all mammalian species for which genome assemblies have been made public. Whole genome shotgun WGS assembly contigs are variable in size. Many are shorter than the endogenous retroviruses ERVs present within them. Whether or not the contig contains the complete ERV is indicated. Whether or not the ERV is intact is indicated. Intact means that all regions are present and there are no obviously deleterious mutations such as frameshift indels or premature stop codons. The status of each region within the ERV is indicated as follows: B Beyond , Entire region is beyond end of the contig; D Defective , Region is fully contained within the contig and contains deleterious mutations; F, Full Region is fully contained within the contig; I Intact , Region is fully contained within the contig and there are no obviously deleterious mutations; P Partial , Region is partially present and the remainder is beyond end of contig. LTR, long terminal repeat. Among the endogenous retroviruses identified in this search, two were found to phylogenetically cluster within the GALV-KoRV-related retrovirus clade Fig. The number of nucleotide positions in the multiple sequence alignments used to generate phylogenies a—e are , , , , and , respectively. The alignment scale is in nucleotides and the total length of each sequence is listed on the right side of the alignment. For comparison, Mastomys WMV shares Mastomys WMV contains the expected open reading frames for gag, pol , and env and all the expected functional motifs are conserved Fig. These include: the proline primer binding site and polyadenylation signal, the major homology region and CCHC zinc finger of Gag, and the protease, reverse transcriptase, and integrase enzymatic active sites Fig. In addition, many of the nucleotide differences in the receptor binding domain compared to WMV are non-synonymous mutations, and the CETTG pathogenicity motif is present in env Fig. The lack of nucleotide differences resulting from genetic drift since integration indicates that this retrovirus integrated into the genome of M. The presence of intact gag, pol , and env genes and functional enzymatic motifs suggests that Mastomys WMV can potentially express infectious viral particles. Two genome assemblies are available for M. Aside from this deletion, Praomys WMV contains mostly uninterrupted reading frames. Taken together, this indicates that this is a defective endogenous retrovirus. It shares We analysed the PiT-1 sequences for all the novel hosts of GALV-KoRV-related retroviruses reported here to determine if they possessed any mutations that might indicate evolutionary adaptation towards non-permissivity. All but one rodent PiT-1 sequence from Z. This analysis also suggested a geographic bias among the hosts of these endogenous retroviruses Fig. Using our targeted search strategy, 27 endogenous retroviruses were identified across 19 species Table 2. These Southeast Asian host species are diverse and include microbats Megaderma lyra and Murina aurata feae , lemurs Galeopterus variegatus , Etruscan shrews Suncus etruscus , pangolins Manis javanica , and treeshrews Tupaia tana. These data indicate that despite not phylogenetically clustering within the GALV-KoRV-related retrovirus clade, a large proportion of gammaretroviruses with receptor binding domain sequence homology to GALV-KoRV-related retroviruses are hosted by Southeast Asian mammals and the host species are predominantly rodents. Evolutionary relationships of novel endogenous gammaretroviruses identified in mammalian genome assemblies via sequence homology with a GALV-KoRV-related retroviral receptor binding domain sequence. Maximum likelihood phylogenies of the a gag , b pol , and c env genes. The GALV-KoRV-related retrovirus clade is shaded in green; branches representing novel endogenous retroviruses reported in this study are shown in orange; branches representing previously reported gammaretroviruses are shown in black. The number of nucleotide positions in the multiple sequence alignments used to generate phylogenies a—c are , , and , respectively. KoRV and its close relatives are viruses of ecological concern, with origins shrouded in mystery, and potentially pathogenic consequences in the event of further cross-species transmission events into humans or other animals. To enhance our understanding of the breadth of the host network of this group of viruses and identify hosts which may require further attention, we searched for previously unreported GALV-KoRV-related retrovirus sequences hidden in the growing expanse of publicly available sequence data. All SRA records in which these retroviral sequences were identified were exomes recently generated from Australian museum specimens Roycroft et al. These rodent species inhabit diverse territories around much of coastal Australia, including Tasmania Fig. However, reported rodent GALV-KoRV-related retroviruses in Australia had previously been limited to the host Melomys burtoni , which ranges in the north and northeastern coast of Australia Aplin et al. This geographic distribution suggests that GALV-KoRV-related retroviruses are widespread across Australia and may infect many more hosts than previously known. Despite this limitation, deleterious mutations in these retroviruses were sufficiently abundant that almost all contigs were found to contain them. This suggests that they are likely endogenous retroviruses and that their integration was not recent. Complete endogenous retroviral genomes from these rodents will help to determine how long ago these integration events occurred. In these cases, they could be included in the same phylogenies Fig. Overlapping but distinct contigs indicate the presence of either multiple retroviruses or duplication of a single integrated retrovirus followed by sequence divergence Johnson Clarifying this will require more extensive sequencing of these rodent genomes. It is also worth noting that five of the seven Australian rodent species are members of the same genus, Pseudomys. Together with the observation that these retroviral sequences are likely not recent integrations, this might suggest that one or more of the endogenous retroviruses represented by these contigs integrated into a common ancestor of these rodents. This may indicate that the analysed gag, pol , and env contigs are derived from different endogenous retroviruses, or that the limited sequence information in these relatively short contigs prevents robust estimation of evolutionary relationships. The latter possibility is suggested by the weak bootstrap support of some internal nodes across the phylogenies Fig. Another important possibility is that some of the contigs may represent ERVs that have arisen through the recombination of different retroviruses. Retroviral recombination is a well understood mechanism of retroviral diversification Negroni and Buc For example, a sub-clade of Type D betaretroviruses which includes primate and bat retroviruses arose through a recombination event that joined a gammaretroviral env region to a betaretroviral gag-pol region Hayward et al. Both endogenous retroviruses appear to be very recently integrated into the M. Mastomys WMV has a fully intact proviral genome. All canonical genes are present in Mastomys WMV, with uninterrupted open reading frames and the conservation of expected functional motifs Fig. The presence of multiple non-synonymous mutations in the receptor binding domain is consistent with ongoing selective pressures in different hosts following the divergence of WMV and Mastomys WMV from their common ancestor. Mastomys WMV is present in one of two available genome assemblies for M. If rodents are the major hosts of this clade of retroviruses and responsible for many interspecies transmission events, then the identification of potentially infectious rodent GALV-KoRV-related retroviruses fills an important gap in supporting this notion. Mastomys WMV may reflect a similar case to KoRV-A and cMWMV in which the virus, being recently integrated, is in the process of endogenization and fixation in the gene pool while still producing functional, infectious retroviral particles. Praomys WMV may also be in the process of undergoing endogenization and fixation in its murine host; however, the identified copy is clearly no longer functional since it has a deletion of nt relative to WMV in the env gene that would prohibit this endogenous retrovirus from generating infectious viral particles. The high degree of nucleotide identity of these African retroviruses to WMV suggests recent transmission. Rodents of various species have been documented as pests aboard ships Song et al. There are shipping routes across the Indian Ocean connecting Southeast Asia, Australia, and East Africa, and it seems reasonable to speculate that a rodent stowaway carrying an infectious WMV variant might explain these findings. While the gag and pol genes of these endogenous retroviruses are widely dispersed throughout the gammaretroviral phylogeny Fig. These are hosted by Asian mice, some of which are now globally distributed. An important caveat here is the possibility of sampling bias among the genomes represented in the public datasets we queried influencing the apparent abundance of GALV-KoRV-related retroviruses in Southeast Asian species relative to other locations. Mammalian species from some geographical regions are likely to be better represented among genome datasets than others. Future analyses that include a larger, more comprehensive, and geographically unbiased collection of reference mammalian genomes may clarify this possibility. Intriguingly, the new report by Mottaghinia et al. Mottaghinia et al. Here we report an additional recent endogenization in the African rodent M. Further taxa screening is almost certain to reveal further details of this unfolding story. The use of PiT-1 for viral entry requires the presence of a permissive protein sequence motif Schneiderman et al. It is reasonable to speculate that evolutionary pressure from PiT-1 receptor-utilizing retroviruses may lead to host adaptations to prevent viral entry, such as through mutations in receptor sequence motifs important for viral envelope interactions. All of the novel rodent hosts except Z. It is unknown whether the codon deletion in Z. Sustained evolutionary pressure that leads to host adaptations to prevent viral infection may suggest that rodents have contended with GALV-KoRV-related retroviruses for a significant evolutionary timeframe. While it is not possible to accurately estimate the integration times of these short contigs without additional sequence data, other ERVs with similarly extensive mutations have been calculated to have integrated at times on the scale of millions of years ago Martins et al. In combination with the high percentage nucleotide identity with the Southeast Asian WMV, this suggests a much more recent transmission, and therefore integration time, than that for the Australo-Papuan rodents. If we consider human-mediated shipping as a means of transmission of retroviruses, it is reasonable to postulate that Mastomys WMV and Praomys WMV integrated as recently as within the last several hundred years. While other mammals are clearly susceptible to infection with these gammaretroviruses, such transmission events may be incidental to the main thread of gammaretroviral divergence and evolution in rodent hosts. Bats are a potential exception. Nucleic acid evidence reveals the presence of HPG or its very close relatives in a number of fruit bat species including P. Serological evidence further indicates the presence of the HPG sub-clade in P. The GALV-KoRV-related retroviruses of pteropid bats form a monophyly that does not contain any known rodent viruses, including the ones reported in this study Hayward et al. This indicates that the HPG sub-clade has become well adapted to circulation among, and transmission between, different bat species. A hypothetical timeline for the spread of GALV-KoRV-related retroviruses could feasibly involve ancestral origins within rodent hosts in the vicinity of mainland Asia or Southeast Asia, with divergence from other gammaretroviral lineages that currently include murine leukemia viruses SI Figure 8A hosted by Asian rodents. Since arrival in Australia, this clade may have been transmitted back and forward between Australia and Southeast Asia by various natural and incidental hosts leading to the eventual integration of WMV variants in the genomes of Australo-Papuan rodents such as Melomys spp. Following arrival in Australia, one lineage that now includes HPG was transmitted into Australian fruit bats Hayward et al. The collective ranges of these species extend from Southeast Asia down through to South Australia. Similarly, the direct retroviral ancestor of KoRV was transmitted to koalas from another species, likely a rodent but possibly some other Australian mammal such as a bat. This hypothetical timeline is depicted in Fig. An important limitation of this study is the lack of breadth of available sequence data. This is somewhat surprising considering that such large amounts of data already exist, but it is important to consider that for most species, data exist from a very limited number of individuals, and for many species no data exist at all. We might compare it to the analogy of trying to find a very specific type of human virus by analysing genomic and transcriptomic data from just a few people. The chances of success might be slim. While over recent years we have been discovering new host species Simmons et al. There may yet be any number of important animal species under threat from this group of cancer-causing viral pathogens. The iterative search strategy employed in this study utilized a short nt sequence representing the receptor binding domain of a GALV-KoRV-related retrovirus. It is important to note that some retroviruses are generated as the product of a recombination event between different, sometimes distantly related retroviruses Negroni and Buc , Hayward et al. The contigs generated from Australian rodent exomes represent highly stringent, consensus short-read assemblies. As such, it is possible that one or more contig assemblies could be generated from reads derived from multiple discrete ERVs. Future genome assemblies generated for these Australian rodents may help clarify this issue. Regarding the potentially infectious nature of Mastomys WMV, it is important to emphasize that no modelling or infectivity experiments were performed, and as such infectivity has not been confirmed. While no obviously deleterious mutations such as frameshifting or stop codon mutations are present in Mastomys WMV, and all key sequence motifs are present and intact, other mutations such as non-synonymous mutations may have rendered this ERV non-infectious. Experimental approaches such as those conducted for HPG Hayward et al. The identification of numerous rodents as novel hosts of GALV-KoRV-related retroviruses widens our understanding of the host range of this clade of oncogenic viruses. It also highlights that our knowledge remains relatively limited. Many further animal species may already host these viruses or be susceptible to infection. The finding of GALV-KoRV-related retroviruses in African rodents, and the expanding association with rodent hosts more generally, demonstrates the transmissibility of these viruses and implicates rodents as an important host reservoir. Since GALV-KoRV-related retroviruses are associated with animal disease, future studies should seek to determine which animals could potentially become infected with them. Transmission of GALV-KoRV-related retroviruses from rodents or bats into animals of domestic, economic, or ecological importance could have dire consequences, as it has had for koalas. This is because gammaretroviruses are known to undergo recombination Bartosch et al. To identify previously unreported GALV-KoRV-related retroviruses within publicly accessible datasets, it was necessary to employ a strategy that would distinguish the target retroviral sequences from non-target retroviral sequences that would appear in our search results due to the presence of conserved sequence regions. Additionally, searches with large sequences, such as the complete KoRV genome, are computationally intensive and time consuming to run on public servers Karasikov et al. Mammalian SRA records are too large and numerous to make comprehensive local analyses practical for all researchers. To account for these challenges, we used an iterative search strategy where our initial search query employed a short sequence from the least conserved region of the retroviral genome, the receptor binding domain within the env gene of a GALV-KoRV-related retrovirus. The rationale for this approach is that since there is a large degree of sequence diversity in the receptor binding domain, even among closely related retroviruses Han et al. Searches were performed for high-level taxa representing the breadth of the class Mammalia. To estimate the evolutionary relationships between the endogenous retroviral sequences identified in genome assemblies and SRA with known gammaretrovirus, we conducted phylogenetic analyses. Endogenous retroviruses were extracted from genome assembly contigs by delineation of their long-terminal repeats as described previously Hayward et al. The gag, pol , and env genes were identified and annotated by sequence alignment against known gammaretrovirus genes using MUSCLE followed by manual curation. Contigs derived from Pseudomys delicatulus env and P. Ambiguous regions of the alignments were removed using Gblocks Castresana Maximum likelihood trees were then inferred using the best-fit model in CLC with bootstrap replicates. Trees were visualized with MEGA We thank Edward C. Holmes and Mark Ziemann for their kind assistance and input in drafting the manuscript. We gratefully acknowledge the contribution of the Victorian Operational Infrastructure Support Program, received by the Burnet Institute. GenBank accessions for the publicly available sequence data used in this study are listed in Tables 1 and 2 , and SI Table 4. This section collects any data citations, data availability statements, or supplementary materials included in this article. As a library, NLM provides access to scientific literature. Virus Evol. Find articles by Joshua A Hayward. Find articles by Shuoshuo Tian. Find articles by Gilda Tachedjian. Published by Oxford University Press. Open in a new tab. Similar articles. Add to Collections. Create a new collection. Add to an existing collection. Choose a collection Unable to load your collection due to an error Please try again. Add Cancel. SRX c. Roycroft et al. SRX d. SRX e. SRX f. SRX g. Kumon et al.

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