Amplification of E2 gene was performed using AccuPrime Taq DNA Polymerase High Fidelity according to the manufacturer’s instruction (Invitrogen). The obtained cDNA was served as template to amplify the full-length E2 gene using forward primer CSFV-E2-F: 5′-GGYRAATATGTGTGTGTWAGACC-3′ and reverse primer: CSFV-E2-R: 5′-TGGTCTTRACTGGRTTGTTRGTC-3′. Briefly, total RNA of the infected kidney tissue was extracted using Trizol reagent (Ambion, Carlsbad, CA) according to the manufacturer’s instruction and subsequently reverse transcribed with Super Script III first strand cDNA synthesis kit (Invitrogen, Carlsbad, CA) using random primer. Detection of wild-type CSFV and other virusesĬSFV in the specimen was detected by amplification of full-length E2 gene with RT-PCR. The kidney specimen was collected from one dead piglet and sent to our laboratory for diagnosis. All these piglets in this farm were imported from the neighboring Guangxi province. Furthermore, the complete genome sequence of this CSFV isolate was determined and analyzed.Īn outbreak of suspected CSF in a pig farm in Guangdong province was reported in 2011, in which 42 of 50 piglets around 30-day old showed clinical signs resembling CSF, and 8 piglets died. In the present study, a sub-subgenotype 2.1c CSFV was isolated from the kidney of a CSFV-infected piglet and adapted to cell culture with high titer following 46 times of continuous passages in PK-15 cells. Most recently, sub-subgenotype 2.1d was identified from outbreaks in Shandong, Jilin, Heilongjiang and Jiangsu Provinces, indicating the presence of genetically divergent subgenotype 2.1 viruses in China. Recent reports showed that a novel CSFV sub-subgenotype, 2.1c, was characterized from CSF outbreaks in Hunan and Guangdong provinces in 2011. reported that 34 out of 35 CSFV isolates collected from Southeast China between 20 were sub-subgenotype 2.1b. Further analysis showed that subgenotype 2.1 isolates could be segregated into sub-subgenotypes 2.1a and 2.1b. In China, subgenotypes 2.1, 2.2, 2.3, 1.1, and 3.4 were reported with the first two subgenotypes causing the most CSF outbreaks in the country. It was reported that CSFV circulating in Europe belong to genotype 2 and isolates from South America belong to genotype 1. CSFV isolates can be classified into 3 genotypes and 11 subgenotypes (1.1–1.4 2.1–2.3 3.1–3.4) based on the 5′UTR (150 nucleotides), E2 (1 nt), and NS5B (409 nt). CSF virus is a member of the genus Pestivirus within the family Flaviviridae, which is an enveloped virus harboring a single-strand positive-sense RNA genome. As an OIE notifiable disease, CSF usually causes significant economic losses in pig industries. Taken together, a field CSFV strain GD53/2011 was isolated, fully sequenced, and adapted to high growth titer in PK-15 cells, which might be suitable for future studies on CSFV infection, replication, and vaccine development.Ĭlassical swine fever (CSF) is a highly contagious swine disease with high morbidity and mortality, featuring symptoms of hemorrhagic fever and immunosuppression. Full-length protein sequence comparison of GD53/2011 with other 2.1 sub-subgenotype isolates showed that Core and NS5A, rather than E2, are more genetically variable. Furthermore, E2 gene sequences at passages 6, 15, 25, 35, and 46 was found identical to that in the original tissue, indicating that the E2 gene was stable during CSF virus adaptation in PK-15 cells. Sequence comparison revealed that the E rns gene at passages 6, 15, and 25 of GD53/2011 was identical to that in the original tissue, but one amino acid substitution (S476R) was detected at passages 35 and 46. After adaptation in PK-15 cells, the titer of GD53/2011 was significantly increased from 10 3.39 TCID 50/ml at passage 6 (F6) to 10 8.50 TCID 50/ml at passage 46 (F46) with the peak titer obtained at 48 h post-inoculation. To further understand the replication characteristics, GD53/2011 was subsequently adapted in PK-15 cells, and its full-length genome was sequenced. Phylogenetic analysis based on the full-length E2 gene sequence revealed that this isolate belongs to CSFV sub-subgenotype 2.1c. This study reports the isolation and characterization of a field CSF virus named GD53/2011 from pig kidney tissue collected during a CSF outbreak in Guangdong province, China. Classical swine fever (CSF) still causes substantial economic losses in the pig industry in China.
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The result is an extraordinary, deeply revealing portrait of Australian crime from the 60s to the 2010s – a portrait of carnage. ASPELUND: It is with deep sadness that the family of Jack Karlson Aspelund announce his passing at River Heights Lodge on May 24, 2019, at ninety-one years. Zelig-like, Karlson had been in the background of many of the most notorious incidents in late-twentieth century Australian crime: co-author of infamous prison-playwright Jim McNeil early collaborator of Charles Billich, one of Australia’s most commercially successful artists associate of hitman Christopher Dale Flannery (Mr Rent-a-Kill).īut most shockingly of all, Karlson’s life story turned out to be the way into cracking the mystery behind a number of unsolved murders, by two serial killers. But during their conversations over the following months, a more complex and darker picture than that of the baritone larrikin began to emerge. When someone named Jack Karlson called Mark Dapin out of the blue, though, Dapin hadn’t heard of him. For Elon Musk, Twitters owner, a Fox News-style show on the platform could cause problems. The Guardian has called it ‘perhaps the pre-eminent Australian meme of the last 10 years’. Tucker Carlson announced his next move: Hes launching a show on Twitter. Whether you know it as the ‘succulent Chinese meal’ video (as it appears on T-shirts) or ‘democracy manifest’ (as it’s taught in schools), chances are you’re one of the millions of Australians who’ve seen Jack Karlson getting arrested outside a Brisbane Chinese restaurant in 1990. Democracy Manifest Man Has Become A Talented Painter And Is Now Taking Submissions Jack Karlson has started doing paintings of his infamous arrest but says he'll do anything that someone suggests. An extraordinary true crime cult classic in the making. Jack Karlson has started doing paintings of his infamous arrest but says he'll do anything that someone suggests. I graduated in medicine from Manchester University in July 2012. But when out of the blue this Falstaff figure called author Mark Dapin thirty years later, it emerged his story went to the heart of the Australian underworld over decades, connecting a celebrated prison playwright, Mr Rent-a-Kill, and two uncaught serial killers. I am a surgeon interested in musculoskeletal health and disease. He even cameoed in a popular music video. There are t-shirts, stubbie holders, wine, pins, magnets and mugs quoting his rant. Online, the one-minute video of his dramatic public defence has become legendary. 21.Millions have been entertained by the viral video of a man being arrested after a ‘succulent Chinese meal’. To this day, Jack Karlson refuses to admit to using a stolen credit card at a Brisbane Chinese restaurant. No guarantee, warranty or representation of any kind is made regarding the completeness or accuracy of descriptions or measurements (including square footage measurements and property condition), such should be independently verified, and Compass expressly disclaims any liability in connection therewith. When someone named Jack Karlson called Mark Dapin out of the blue, though. and/or other countries.Ĭorporate Responsibility, Privacy & Legal Notices: Compass is a licensed real estate broker, licensed to do business as Compass RE in Delaware, Idaho, New Jersey, Pennsylvania and Tennessee, Compass Real Estate in Washington, DC, Wyoming and Idaho, Compass Realty Group in Missouri and Kansas, and Compass South Carolina LLC in South Carolina. Do Not Sell or Share My Personal Information, Terms of Service, Privacy Center, Scam Avoidance, Responsible Disclosure, Compass is an E-Verify employer, Notice for California Applicants, California COVID-19 Rules of Entry, and Your CA Privacy RightsĬOMPASS, the Compass logo, and other various trademarks, logos, designs, and slogans are the registered and unregistered trademarks of Compass, Inc. To be fair, his first half deliveries had been disappointing at a stage when the visitors were dominating, so they felt even more wasteful. James Bree had the last laugh against Fulham and perhaps should’ve stolen all the glory (see the next talking point). James Bree provided the assist for Elijah Adebayo’s equaliser and then made a point to celebrate in front of a fan who’d been on his back. To put it into context – and a caveat that Luton are now a better side than in their first to Championship campaigns – he is well within range of comfortably excelling beyond his striking predecessor, James Collins, who bagged 14 in 46 games in his debut second tier season, then 13 in 44 last term.Īnd, as an added bonus, Luton have never lost when Adebayo has scored. This time last year, he was netting in a 2-1 League Two win for Walsall over Bolton, the seventh of ten goals in 28 bottom tier appearances. Now, in the Championship, he’s bang on for a goal every other game, with ten goals in 20 outings. The 23-year-old is now one of only five forwards in that club, with Fulham’s Mitrovic (22), Blackburn’s Ben Brereton-Diaz (17), Bournemouth’s Dominic Solanke (16) and Swansea’s Joel Piroe (11). It was only going to end one way □ | #COYH /Kh91WcUom9- Luton Town FC December 12, 2021 His headed equaliser against Fulham – a tremendously under-appreciated skill against his former side – moved him into double figures for the season. The striker has not yet had a full calendar year as a Championship player and he’s already mixing it with the boys of the goal-getting department. When you’ve got that, and talent and some genetic attributes, then he’s some force.” The good thing about him is he’s just a humble kid that wants to work hard. And we won’t let him rest on his laurels. “That’s a great thing for us and we will reap the benefit. He’s going to get better, he’s not going to get any worse. “He doesn’t get paid as much as Mitrovic,” said the Town boss, adding: “Pound for pound, there’s not a striker like him in the Championship and that’s what we’re proud of. For Manager Nathan Jones, he’s already one of the best two number nines in the division, the other being Aleksander Mitrovic. He’s the definition of a precocious talent and getting better all the time. Photo by Liam Smithīe honest nobody expected Elijah Adebayo to be this good, this quick, did they? We kept winning the ball quite high up and we were creating chances.” Pelly-Ruddock Mpanzu jumps on goalscorer Elijah Adebayo. They did have a few spells but overall, we were right at it. “After the goal, the gaffer changed the shape a bit and it worked well for us. I think everyone, dealt well with what they had because they’re a good team. “We are disappointed we didn’t get a clean sheet. I think the reaction from us was very good. It’s something we need to go away and look at to make sure it doesn’t happen again. “As a defender, conceding from a set-piece is disappointing and we should be doing better with that. Overall, we had the better chances and I think we played well and maybe deserved a bit more,” he said. “We conceded from a set play and maybe should’ve done better than that. So a point was the least Luton deserved, but Burke’s reaction to restricting the output of such a talented and expensive side speaks volumes to the the evolution process that is taking place at Luton. I was far away from it and couldn’t see much, but it did look like a foul.” Because it’s inside the box, it gives the referee and decision to make and maybe it wasn’t the right one. And, if the referee hadn’t bottled given a penalty for Joe Bryan’s barge in the back of Fred Onyedinma late on then it could’ve been even better than a point.īoss Nathan Jones said it was a clear penalty, while defender Reece Burke added: “From my view, I think if it’s outside the box, it may be given. I expected nothing, and got an impressive performance. Pessimists and realists alike might’ve been hard pressed to predict anything favourable for Luton. A top performance but disappointment at the result is a positiveįulham arrived at Kenilworth Road as the Championship winners elect on account of averaging two goals a game, having the deadliest striker the division, in Aleksander Mitrovic, and one of the most miserly defences.Kal Naismith pleads his case to referee Thomas Bramall with Aleksander Mitrovic rolling around on the Kenilworth Road turf. Here are six spoonfuls of sugar we can take from holding the Championship leaders. Luton held Fulham to a point at Kenilworth Road on Saturday after coming from a goal behind with a goal from top scorer Elijah Adebayo. |
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