The FBI’s Atlanta Field Office has announced the seizure of several online criminal marketplaces providing pirated versions of popular video games.
In addition to the seizure, the FBI has dismantled the infrastructure of these websites.
For more than four years, the websites contained pirated copies of highly anticipated video games days or weeks before their official release date.
Between February 28th, 2025, and May 28th, 2025, records indicate a total of 3.2 million downloads occurred on these sites from the most used download service. These actions resulting in an estimated loss of $170 million (€146m).
As part of the actions, the FBI has obtained authorisation to seize the domain of multiple sites, to include nsw2u.com, nswdl.com, game-2u.com, bigngame.com, ps4pkg.com, ps4pkg.net, and mgnetu.com. These domains hosted and facilitated access to the pirated video games. Anyone visiting these sites will now view a seizure banner that notifies them the domain has been seized by federal authorities.
The FBI has thanked the Dutch FIOD for their assistance in this investigation.
They went viral, amassing more than 1m streams on Spotify in a matter of weeks, but it later emerged that hot new band the Velvet Sundown were AI-generated – right down to their music, promotional images and backstory.
The episode has triggered a debate about authenticity, with music industry insiders saying streaming sites should be legally obliged to tag music created by AI-generated acts so consumers can make informed decisions about what they are listening to.
Initially, the “band”, described as “a synthetic music project guided by human creative direction”, denied they were an AI creation, and released two albums in June called Floating On Echoes and Dust And Silence, which were similar to the country folk of Crosby, Stills, Nash & Young.
Things became more complicated when someone describing himself as an “adjunct” member told reporters that the Velvet Sundown had used the generative AI platform Suno in the creation of their songs, and that the project was an “art hoax”.
The band’s official social media channels denied this and said the group’s identity was being “hijacked”, before releasing a statement confirming that the group was an AI creation and was “Not quite human. Not quite machine” but living “somewhere in between”.
Several figures told the Guardian that the present situation, where streaming sites, including Spotify, are under no legal obligation to identify AI-generated music, left consumers unaware of the origins of the songs they’re listening to.
Roberto Neri, the chief executive of the Ivors Academy, said: “AI-generated bands like Velvet Sundown that are reaching big audiences without involving human creators raise serious concerns around transparency, authorship and consent.”
Neri added that if “used ethically”, AI has the potential to enhance songwriting, but said at present his organisation was concerned with what he called “deeply troubling issues” with the use of AI in music.
Sophie Jones, the chief strategy officer at the music trade body the British Phonographic Industry (BPI), backed calls for clear labelling. “We believe that AI should be used to serve human creativity, not supplant it,” said Jones.
“That’s why we’re calling on the UK government to protect copyright and introduce new transparency obligations for AI companies so that music rights can be licensed and enforced, as well as calling for the clear labelling of content solely generated by AI.”
Liz Pelly, the author of Mood Machine: The Rise of Spotify and the Costs of the Perfect Playlist, said independent artists could be exploited by people behind AI bands who might create tracks that are trained using their music.
She referred to the 2023 case of a song that was uploaded to TikTok, Spotify and YouTube, which used AI-generated vocals claiming to be the Weeknd and Drake. Universal Music Group said the song was “infringing content created with generative AI” and it was removed shortly after it was uploaded.
It is not clear what music the Velvet Sundown’s albums were trained on, with critics saying that lack of clarity means independent artists could be losing out on compensation.
Pelly said: “We need to make sure that it’s not just pop stars whose interests are being looked after, all artists should have the ability to know if their work has been exploited in this way.”
For some, the appearance of the Velvet Sundown is the logical next step as music and AI combine, while legislation is fighting to keep up with a rapidly changing musical ecosystem.
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Jones said: “The rise of AI-generated bands and music entering the market points to the fact that tech companies have been training AI models using creative works – largely without authorisation or payment to creators and rights-holders – in order to directly compete with human artistry.”
Neri added that the UK has a chance to lead the world in ethical AI adoption in music but said there needed to be robust legal frameworks that “guarantee consent and fair remuneration for creators, and clear labelling for listeners”.
“Without such safeguards, AI risks repeating the same mistakes seen in streaming, where big tech profits while music creators are left behind,” he added.
Aurélien Hérault, the chief innovation officer at the music streaming service Deezer, said the company uses detection software that identifies AI-generated tracks and tags them.
He said: “For the moment, I think platforms need to be transparent and try to inform users. For a period of time, what I call the ‘naturalisation of AI’, we need to inform users when it’s used or not.”
Hérault did not rule out removing tagging in future if AI-generated music becomes more popular and musicians begin to use it like an “instrument”.
Deezer recently told the Guardian that up to seven out of 10 streams of AI-generated music on the platform are fraudulent.
At present, Spotify does not label music as AI-generated and has previously been criticised for populating some playlists with music by “ghost artists” – fake acts that create stock music.
A spokesperson for the company said Spotify does not prioritise AI-generated music. “All music on Spotify, including AI-generated music, is created, owned and uploaded by licensed third parties,” they said.
Travel the universe with Dr. Ethan Siegel as he answers the biggest questions of all.
Since the 1800s, energetic particles have probed the fundamental nature of matter.
Rutherford’s gold foil experiment showed that the atom was mostly empty space, but that there was a concentration of mass at one point that was far greater than the mass of an alpha particle: the atomic nucleus. By observing that some of the emitted, radioactive particles bounced back, or ricocheted off, in a different direction than they were emitted in, Rutherford was able to demonstrate the existence of a compact, massive nucleus to the atom.
Credit: Chris Impey
By bombarding matter with other particles, we probe their internal structures.
The production of matter/antimatter pairs (left) from two photons is a completely reversible reaction (right), with matter/antimatter annihilating back to two photons. This creation-and-annihilation process, which obeys E = mc², is the only known way to create and destroy matter or antimatter. If high-energy gamma-rays collide with other particles, there is a chance to produce electron-positron pairs, diminishing the gamma-ray flux observed at great distances.
Credit: Dmitri Pogosyan/University of Alberta
At still greater energies, we create new quanta via Einstein’s E=mc².
By taking a hot air balloon up to high altitudes, far higher than could be achieved by simply walking, hiking, or driving to any location, scientist Victor Hess was able to use a detector to demonstrate the existence and reveal the components of cosmic rays. In many ways, these early expeditions, dating back to 1912, marked the birth of cosmic ray astrophysics.
Credit: VF Hess Society, Schloss Pöllau/Austria
Early experiments with cosmic rays first revealed heavy, unstable Standard Model particles.
The first muon ever detected, along with other cosmic ray particles, was determined to be the same charge as the electron, but hundreds of times heavier, due to its speed and radius of curvature. The muon was the first of the heavier generations of particles to be discovered, dating all the way back to the 1930s.
Credit: P. Kunze, Zeitschrift für Physik, 1933
Then particle accelerators and colliders arrived, revealing nature’s secrets at still higher energies.
Bubble chamber tracks from Fermilab, revealing the charge, mass, energy, and momentum of the particles and antiparticles created. This recreates similar conditions to what was present during the Big Bang, where matter and antimatter can both be readily created from pure energy. At the highest energies, all particles and antiparticles can be created, but at energies corresponding to “only” a temperature of ~10 billion K or so, electron-positron pairs can still be spontaneously created.
Credit: Fermi National Accelerator Laboratory/DOE/NSF
As the energy frontier progressed, more and more of the Standard Model was uncovered.
This chart of particles and interactions details how the particles of the Standard Model interact according to the three fundamental forces that quantum field theory describes. When gravity is added into the mix, we obtain the observable Universe that we see, with the laws, parameters, and constants that we know of governing it. However, many of the parameters that nature obeys cannot be predicted by theory, they must be measured to be known, and those are “constants” that our Universe requires, to the best of our knowledge.
Finally, in the 2010s, the Higgs boson was discovered at the Large Hadron Collider (LHC), completing the Standard Model.
The first robust, 5-sigma detection of the Higgs boson was announced a few years ago by both the CMS and ATLAS collaborations. But the Higgs boson doesn’t make a single ‘spike’ in the data, but rather a spread-out bump, due to its inherent uncertainty in mass. Its mass of 125 GeV/c² is a puzzle for theoretical physics, but experimentalists need not worry: it exists, we can create it, and now we can measure and study its properties as well. Direct detection was absolutely necessary in order for us to be able to definitively say that.
Credit: CMS Collaboration/CERN
Many hope to build new, more powerful colliders, attempting to unearth additional secrets about reality.
Deep underground, this tunnel is part of interior workings of the Large Hadron Collider (LHC), where protons pass each other at 299,792,455 m/s while circulating in opposite directions: just 3 m/s shy of the speed of light. Particle accelerators like the LHC consist of sections of accelerating cavities, where electric fields are applied to speed up the particles inside, as well as ring-bending portions, where magnetic fields are applied to direct the fast-moving particles toward either the next accelerating cavity or a collision point.
Credit: Maximilien Brice and Julien Marius Ordan, CERN
It could be a linear collider: probing heavy, unstable particles exquisitely.
Although there are many novel proposals for new particle colliders, including in China, at CERN, and at Fermilab, the question of whether to build a circular machine, a linear lepton collider, or to pursue a novel muon collider all remain options on the table. In an ideal world, we’d get a linear machine to study the Higgs and the electroweak phase transition with great precision, and then a circular machine to collide hadrons at even higher energies. But funding, political realities, and popular opinion will also play a major role in determining what decisions get made.
Credit: Rey Hori/KEK
It could be a circular collider, progressing farther than ever into the energy frontier.
The Future Circular Collider (in blue) would overlap slightly with the current Large Hadron Collider, but requires an additional ring (and tunnel) somewhere upward of 80 km in circumference: dwarfing the LHC’s current 27 km circumference. Bigger tunnels and stronger magnets are needed for a more energetic hadron collider, with the FCC proposing ~16 T magnets, approximately double the LHC’s current magnet strength.
Credit: CERN / Big Think
Someday, we may even build a collider around the Earth: thousands of times as powerful as the LHC.
This illustration shows a hypothetical ring around the Earth, which could represent a particle accelerator even larger than the Earth’s circumference. With approximately ~1500 times the radius of the Large Hadron Collider, such an accelerator, even with only slightly more advanced magnet technology, would be thousands of times more powerful. A particle accelerator that was merely a factor of ~10 more powerful than the LHC could potentially shed tremendous light on the matter-antimatter asymmetry puzzle.
Credit: Adrian Mann/aerospace illustration/bisbos
At some point, however, there will be a limit to whatever energies colliders can pragmatically reach.
In this artistic rendering, an active, supermassive black hole whose jet points at us (a blazar) is accelerating protons to extreme energy, producing pions as daughter particles, which in turn produce neutrinos and gamma rays. Extreme events in energy are thought to be generated by processes occurring around the largest supermassive black holes known in the Universe when they’re actively feeding. The energies of these cosmic rays vastly exceed those achieved in terrestrial accelerators.
Credit: IceCube collaboration/NASA
However, the Universe creates cosmic rays exceeding ~1011 GeV: millions of times the LHC’s maximum energy.
The energy spectrum of the highest energy cosmic rays, by the collaborations that detected them. The results are all incredibly highly consistent from experiment to experiment, and reveal a significant drop-off at the GZK threshold of ~5 x 10^19 eV. Still, many such cosmic rays exceed this energy threshold, indicating that the heaviest cosmic rays are likely heavy nuclei, rather than the more common bare proton.
Credit: M. Tanabashi et al. (Particle Data Group), Phys. Rev. D, 2019
After the last collider has finished, rare, ultra-energetic cosmic rays will continue revealing the Universe’s secrets.
In May of 2021, the second most energetic cosmic ray ever detected struck Earth, producing a shower of particles detected on the ground by the Telescope Array Collaboration. These particles achieve energies more than a million times greater than the maximum LHC energy, such that after humanity has built our last collider, the energy frontier will still be accessible from space, albeit extremely rarely.
Love heartbreakingly clashes with economic difficulties in Ico Costa’s third feature, shot on location in Mozambique and featuring non-professional actors. The film opens on a gentle moment between Domingos (Domingos Marengula) and Neusia (Neusia Emídio Guiamba), their figures wrapped in the velvety half-light of the early dawn. Lying in bed together, the young couple sleepily talk of mundane trivialities but their tender gaze seems to speak louder than words; locked in each other’s arms, the pair will soon spend the rest of the film apart.
Unsatisfied with the meagre earnings he makes from washing cars, Domingos leaves his home town for the dangerous goldmines in the north of the country. When he enters this precarious trade, the camera trails behind his footsteps as he passes through rocky terrains and makeshift tents. It’s an interesting stylistic choice that seeks to emphasise both the geographical specificity and the lethal risks of Domingos’s environment, as well as his bond with fellow workers. When seen in group compositions, the men share a moving rapport that can’t always resist the callous indifference of an exploitative industry.
Nevertheless, with Domingos largely filmed from behind, much of the intimacy seen in the striking first scene is lost. The camera simply gazes at him and his circumstances; we don’t get to know anything of his internal world. In comparison, Neusia, who stays behind, is shot in more dynamic closeup; though her scenes are so far and few between that she does not come off as a fully fledged character either. Costa certainly has a keen eye for the landscape and the working-class milieu of Mozambique, but his purely observational style reduces Gold Songs to a detached ethnographic portrait rather than a properly absorbing drama.
The universe holds endless mysteries, and today’s most powerful telescopes are helping us explore them like never before. These advanced instruments drive the progress of modern astronomy, using radio, infrared, optical, and X-ray wavelengths to capture light from ancient galaxies, black holes, exoplanets, and other distant objects. Built in extreme environments and equipped with modern technology, they allow scientists to see far into space and look back in time. Each telescope on this list plays a vital role in uncovering how the universe began, how it changes, and what might exist beyond what we know.
Top 10 most powerful telescopes on Earth and in space
1. James Webb Space Telescope (JWST)
Launched in December 2021, the James Webb Space Telescope (JWST) is placed about 1.5 million kilometres away from Earth, at a special spot in space called the Sun–Earth L2 point. This location is stable and perfect for observing deep space without interruptions. JWST looks mainly in infrared light, which helps it see through thick clouds of space dust. This allows it to spot stars and galaxies that formed soon after the Big Bang. Its powerful instruments are so sensitive that they can catch the faintest light from faraway galaxies and even study the atmospheres of planets outside our solar system—possibly helping us find signs of life.
Located in a natural depression in Guizhou, China, FAST is the world’s largest and most sensitive single-dish radio telescope. With a massive 500-metre dish, FAST listens to the universe in radio frequencies—essential for detecting signals from distant pulsars, mapping interstellar hydrogen, and searching for potential extraterrestrial intelligence (SETI). Its sensitivity enables the discovery of otherwise undetectable cosmic phenomena across vast distances.
3. Extremely Large Telescope (ELT)
Under construction atop Cerro Armazones in Chile, the ELT will be the largest optical/infrared telescope ever built, with a 39-metre main mirror composed of 798 hexagonal segments. Its light-gathering power will be 250 times greater than Hubble’s and will provide images 15 times sharper. Scheduled for first light around 2029, ELT is designed to investigate dark matter, black holes, early galaxies, and potentially habitable exoplanets—pushing the limits of what we know about the universe.
4. Giant Magellan Telescope (GMT)
Also rising in Chile’s high desert, the GMT uses seven large mirrors to act as a single, 24.5-metre telescope. It promises image clarity up to ten times better than Hubble, enabling it to see incredibly fine details in distant objects. Scientists hope to use GMT to directly image Earth-like planets, explore galaxy formation, and deepen our understanding of the universe’s accelerated expansion.
5. Thirty Meter Telescope (TMT)
Planned for construction atop Mauna Kea, Hawaii (though delayed due to site access disputes), the TMT will feature a 30-meter segmented mirror, optimised for near-infrared and optical observations. It’s designed to study everything from the formation of the first galaxies to the evolution of black holes and the search for life-supporting exoplanets, offering unmatched resolution in ground-based astronomy.
6. Gran Telescopio Canarias (GTC)
Located on La Palma in Spain’s Canary Islands, GTC is currently the world’s largest single-aperture optical telescope with a 10.4-metre mirror. It’s been instrumental in studying dark energy, stellar explosions (supernovae), and planet formation. Its location—far from city lights and high above sea level—makes it ideal for observing the universe with minimal atmospheric distortion.
7. Atacama Large Millimeter/submillimeter Array (ALMA)
Sitting high in Chile’s Atacama Desert, ALMA consists of 66 movable radio antennas working together as one giant interferometer. By observing the coldest regions of space in millimetre and submillimetre wavelengths, ALMA can peer into dense gas clouds to uncover the birthplaces of stars and planets. It also studies ancient galaxies and the building blocks of life, such as organic molecules.
8. Gemini Observatory (North & South)
Gemini consists of two twin 8.1-metre telescopes—one in Hawaii (Gemini North) and the other in Chile (Gemini South). Together, they provide full-sky coverage. Equipped with adaptive optics and powerful spectrographs, Gemini can capture clear, detailed images of distant galaxies, stellar nurseries, and gamma-ray bursts. Its versatility makes it one of the most productive observatories in modern astronomy.
9. Chandra X-ray Observatory
Launched in 1999, NASA’s Chandra remains one of the most important telescopes for observing the universe in X-rays, a high-energy form of light. It specialises in studying extreme environments—such as the hot gas swirling around black holes, exploding stars, and neutron stars. Chandra’s precision has helped us understand the life cycles of stars and the structure of galaxy clusters.
Located in New Mexico, USA, the MROI uses a technique called interferometry, where light from multiple smaller telescopes is combined to simulate the resolution of a much larger one. This approach yields ultra-high-resolution images of binary star systems, stellar surfaces, and debris disks around young stars—objects typically too fine to resolve using single-mirror telescopes.
Why these telescopes are essential tools in modern astronomy
These telescopes represent the pinnacle of astronomical technology. Their large apertures allow them to gather light from the farthest corners of the universe, enabling us to look back in time. Each observatory focuses on specific wavelengths—infrared, radio, X-ray, or optical—uncovering different layers of cosmic phenomena. Technologies like adaptive optics and interferometry enhance clarity, letting scientists image distant galaxies, exoplanets, black holes, and supernovae with astonishing precision.
India’s growing role in modern astronomy
Though not featured in the top‑ten global list, India contributes significantly to modern astronomy infrastructure:
Major Atmospheric Cherenkov Experiment (MACE): Operational from Hanle, Ladakh, at ~4,500 m altitude, MACE is one of the world’s highest gamma-ray telescopes. Commissioned in 2021 and inaugurated in 2024, it advances research in cosmic rays and fundamental physics.
Devasthal Optical Telescope (DOT): Located at ARIES in Uttarakhand, the 3.6-metre DOT is Asia’s largest optical telescope. Commissioned in 2016, it supports advanced imaging and spectroscopy and plays a critical role in regional space research.
These ten telescopes are among the most advanced tools ever built for exploring space. From JWST’s deep cosmic gaze to ALMA’s insight into galactic birthplaces, they’re transforming our understanding of the universe. As more such observatories become operational—and with countries like India boosting their astronomical capabilities—the future of space exploration looks brighter than ever.
In a delicious twist, the return coincides with McIlroy ending his 11-year major drought by winning the Masters to complete the career Grand Slam. The Northern Irishman is here wearing the coveted Green Jacket.
No further hype needed. “Yeah, poor Rory, everyone seems to build up the pressure on him being the favourite,” Harrington said.
“But if you want to be at that level the pressure’s always going to be on you.
“Clearly, he knows Portrush very well, he’ll have the support and there’s no doubt we’d love to see an Irish winner.”
But Harrington says McIlroy should maintain some perspective for what could otherwise be an overwhelming week.
“Him going with the Masters’ jacket, I think it’s enough for him to just swan around and wave to the crowds,” said the three-time major winner.
“He doesn’t have to win. The people always want him to win the next major or whatever, but it doesn’t have to be this one.
“I know it would be nice to be Portrush, but he’ll win plenty more majors.”
Regardless of whether Portrush can serve up another domestic fairytale, this will remain a golden period for golf on the island of Ireland. How does Harrington think the sport’s historians will reflect on it in years to come?
“Clearly it’s been unprecedented,” he said. “There’s been a lot of ‘how did we do it?’ You know, I don’t know if you can replicate things like that.
“Everybody’s been trying to find the formula, did we have something special in Ireland? I’m not sure.
“We gained some momentum. We did our thing. I think it’s good for us going forward that we will have players who will believe in themselves.”
They will do so while speculation grows that new ground will be broken by the R&A taking a future Open to Portmarnock in the the Republic of Ireland.
It is another indicator of how far and how quickly golf in this part of the world has moved. “Definitely, that’s a big step,” Harrington said.
“It’s tried for a long time to lose the tag as the British Open; it’s The Open,” Harrington said.
“And it represents everybody, not just the people in Britain, but it represents everybody around the world who plays golf.
“It’s everybody’s Open.” But this week with a discernible Irish hue.
Mike Covell, founder of Entertainment in Motion and recipient of the APEX Lifetime Achievement Award, passed away peacefully on July 11, 2025, at the age of 86. A trailblazer in the in-flight entertainment industry, Covell spent decades shaping how passengers experienced content in the air, elevating airline entertainment from a novelty to a global expectation.
Covell launched Entertainment in Motion in the early 1980s with a single but significant deal: securing airline rights to the film Da. That early success laid the groundwork for a company that would become one of the industry’s premier distributors of films and television to airlines and cruise lines around the world.
Caption: Attending a Post Modern Edit Golf event in 2013 included (from left to right) West Entertainment President Rick Warren, Disney Senior Manager of Non-Theatrical Sales Melinda Meyer-Gilmore, Warner Bros. Vice President of Non-Theatrical Distribution Evelyn Saunders, and Entertainment in Motion Founder Mike Covell (Photo from Eric Silverstein).
In 2007, the Airline Passenger Experience Association (APEX) recognized his contributions by awarding him its highest individual honor.
“Mike Covell is the consummate professional,” said APEX Board Member Stanley Hosford during the ceremony, as noted in the APEX 2007 Annual Report. “He had a vision for this industry long before most others saw its potential.”
From Studio Roots to Global Distribution
Before entering aviation, Covell honed his expertise at Hal Roach Studios and MGM. His experience in Hollywood gave him the tools to navigate licensing, production, and content curation at a time when few had considered how film might translate to aircraft cabins. He started his IFE career at Trans Com, with a keen understanding of both storytelling and operational logistics, he recognized early on the importance of customized content for different markets and audiences.
As Entertainment in Motion grew, Covell established strong partnerships with major studios and international airlines. His leadership guided the company through multiple technological shifts, from film reels and video cassettes to hard drives and digital streaming platforms. He maintained a focus on quality and consistency, helping airlines deliver engaging passenger experiences no matter the flight length or route.
Mike Covell working at Entertainment in Motion in the early 1990s (Photo from Covell Family).
A Voice of Integrity in a Transforming Industry
Throughout his career, Covell was known for his fairness, mentorship, and unwavering professionalism. His calm demeanor and deep knowledge of the field earned him the respect of colleagues across the aviation and media sectors.
“Mike Covell was one of the true architects of our industry,” said APEX Group CEO Dr. Joe Leader. “He created not just a business, but a legacy of storytelling at altitude.”
In an interview following his Lifetime Achievement Award, Covell reflected on the longevity of his career. “I never dreamed this would be a lifelong career,” he said. “But when you love what you do, it doesn’t feel like work.”
Even in his later years, Covell remained engaged with the evolution of airline entertainment, providing insight as the industry expanded into streaming, wireless delivery, and data-driven personalization.
An Influence That Continues to Fly
Memorial photo for Michael Theodore Covell, January 25, 1939 – July 10, 2025 (Photo from Covell Family).
Entertainment in Motion continues to live on as a part of Anuvu after its acquisition fifteen years ago. The legacy of the company remains a light carried forward in the inflight entertainment ecosystem, reflecting the standards and spirit Covell established from the very beginning. His work redefined how entertainment reaches travelers at altitude, leaving an enduring imprint on every screen that lights up between takeoff and landing.
“Mike Covell, our ‘Mr. Hollywood,’ was a mentor, yes, but more profoundly, he was family – a father, a brother, a true friend,” fellow APEX Lifetime Achievement Award Winner and West Entertainment CEO Rick Warren stated. “He was always available to listen and advise. His wisdom and warmth were often shared over a martini and always with a good laugh. We love him dearly, and he will be deeply missed.”
According to an announcement shared by the marketplace, the games are scheduled to go offline on July 17, 2025, at 3 PM CEST and will no longer be available for purchase afterward, though existing owners will still have access to them.
Furthermore, the team’s statement reveals they have no details about discounts, implying that GOG itself doesn’t have much information on the “why” behind the delisting, which, in turn, suggests that the decision to remove the games was Playdead’s and not theirs. Speaking of discounts, as of now, Inside is available on GOG with a steep 90% discount, while Limbo is still being sold at its regular price.
Between 2006 and 2020, eight Burmese individuals were newly diagnosed with HIV in Baoshan, China. The participants included six females and two males, with ages ranging from twenty-one to forty-two years old in 2020, and ethnicities including Lisu, Dai, Han, and Jingpo (Table 1). Most participants were either uneducated or had only received primary education, and all were farmers. The majority were married, but three participants were unmarried. The primary route of HIV transmission was heterosexual contact, with only one male infected through needle sharing (Table 1).
Table 1 Characteristics of Burmese people living with HIV in baoshan, China
Prevalence of known subtypes of HIV strains
The maximum likelihood tree, constructed using near full-length genomes of HIV, showed that among the eight sequences amplified from Burmese PLHIV in Baoshan, only one sequence (YN33F28) clustered with the known subtype C, and one sequence (YN9M24) clustered with the known CRF08_BC, suggesting that they likely belonged to subtype C and CRF08_BC, respectively (Fig. 1). The remaining six sequences did not cluster with any known subtypes, indicating that they might have represented newly formed recombinant strains (Fig. 1). These findings were corroborated by a neighbor-joining tree incorporating reference sequences of all HIV subtypes and CRFs (Figure S1).
Fig. 1
Maximum likelihood tree based on the near full-length genome of HIV. The red triangles represent the sequences amplified in this study, and the black circles represent the similar sequences downloaded from the HIV database. The sectorial shadings in different colors are used to distinguish different HIV subtypes
The Bootscan plot of sequence YN33F28 showed that it was most similar to HIV subtype C, with no recombination breakpoints, further confirming that it was indeed subtype C (Fig. 2A). The Bootscan plot of sequence YN9M24 demonstrated that its recombination structure was consistent with that of CRF08_BC, which was formed by recombination between subtype B and subtype C. It contained three subtype B fragments and three subtype C fragments, separated by five breakpoints (Fig. 2B). These results further confirmed that sequence YN9M24 belonged to CRF08_BC.
Fig. 2
Bootscan plots of HIV sequences of known subtypes. (A) Subtype C; (B) CRF08_BC. In each plot, the lines in different colors represent the reference sequences of different subtypes, and the black arrows indicate the shared breakpoints among different HIV sequences
Recombinant strains formed through second-generation recombination
The maximum likelihood tree revealed that sequences YN36F38, YN35F22, YN34F21, YN7F27, and YN32M22 clustered with known CRFs (CRF82_cpx, CRF86_BC, and CRF178_BC) and a URF (KY406739), respectively. However, these sequences were positioned closer to the root of the phylogenetic tree compared to their corresponding reference sequences, indicating that they were genetically closely related but not identical to the reference sequences. This suggested that they might have been recombinant strains formed through second-generation recombination involving these reference sequences (Fig. 1).
Bootscan analysis further supported this hypothesis. YN36F38 exhibited a recombination structure in the latter half of the genome identical to that of CRF82_cpx, with three shared recombination breakpoints (Fig. 3A). Similarly, YN35F22 shared a recombination structure in the latter half of the genome with CRF86_BC, with five shared recombination breakpoints (Fig. 3B). Meanwhile, YN34F21 and YN7F27 each shared four and three recombination breakpoints with CRF178_BC, respectively (Fig. 3C). YN32M22, on the other hand, shared two recombination breakpoints with KY406739 in both the anterior and posterior regions of the genome (Fig. 4). These findings provided further evidence that these five sequences (YN36F38, YN35F22, YN34F21, YN7F27, and YN32M22) represent recombinant strains resulting from second-generation recombination involving known CRFs or URFs.
Fig. 3
Bootscan plots of second-generation recombinant HIV sequences composed of CRFs. (A) CRF82_cpx; (B) CRF86_BC; (C) CRF178_BC. In each plot, the lines in different colors represent the reference sequences of different subtypes, and the black arrows indicate the shared breakpoints among different HIV sequences
Fig. 4
Bootscan plots of second-generation recombinant HIV sequences composed of URF_BC. In each plot, the lines in different colors represent the reference sequences of different subtypes, and the black arrows indicate the shared breakpoints among different HIV sequences
Maximum likelihood trees constructed from the subregions of these five sequences revealed that their identical fragments clustered with their respective parental sequences (Figure S2). This observation further supports the conclusion that these sequences are second-generation recombinants derived from known CRFs or URFs.
Newly formed URFs
The remaining sequence, YN8F28, did not cluster with any known subtypes, CRFs, or URFs. Instead, it positioned closer to the root of the phylogenetic tree, suggesting that it was genetically distant from known sequences and might have represented newly formed URF (Fig. 1).
Bootscan analysis confirmed that YN8F28 was a recombinant strain derived from HIV subtypes B and C, as well as CRF01_AE, comprising four subtype B fragments, six subtype C fragments, and four CRF01_AE fragments (Fig. 5). Although phylogenetically positioned between CRF82_cpx and CRF83_cpx (Fig. 1), YN8F28 shared no recombination breakpoints with these CRFs (Fig. 5). These findings indicated that YN8F28 was a novel URF arising from complex recombination events involving multiple HIV subtypes.
Fig. 5
Bootscan plots of newly identified HIV URF and known CRFs. In each plot, the lines in different colors represent the reference sequences of different subtypes