During the early phases of the COVID-19 pandemic, concerns were raised that the use of the painkiller ibuprofen may lead to a more severe course of coronavirus disease. As use of ibuprofen and other NSAIDs is widespread, data on their safety is urgently needed to guide clinicians and patients. In the new study, researchers obtained data on all 9,326 Danish residents who tested positive for the SARS-CoV-2 virus between February 27, 2020 and April 29, 2020. Data were available on NSAID use, 30-day mortality, hospitalization, ICU admission, mechanical ventilation, and acute renal replacement therapy. 248 people (2.7%) had filled a prescription for NSAIDs within 30 days of their positive virus test.

The researchers found no association between any of the outcomes and NSAID use. Among NSAID users in a matched cohort who tested positive for the coronavirus, 6.3% (95% Confidence Interval [CI] 3.1-9.4) died, 24.5% (95% CI 18.6-30.4) were hospitalized and 4.9% (95% CI 2.1-7.8) were admitted to ICU. Of those who tested positive for the coronavirus but were not treated with NSAIDs, 6.1% (95% CI 4.4-7.8) died, 21.2% (95% CI 18.1-24.3) were hospitalized, and 4.7% (95% CI 3.2-6.2) were admitted to ICU. None of these differences between groups were statistically significant (mortality Risk Ratio [RR] 1.02, 95%CI 0.57-1.82, p=0.95; hospitalization RR 1.16, 95%CI 0.87-1.53, p=0.31; ICU admission RR 1.04, 95%CI 0.54-2.02, p = 0.90).

“Considering the available evidence, there is no reason to withdraw well-indicated use of NSAIDs during the SARS-CoV-2 pandemic,” the authors say. “However, the well-established adverse effects of NSAIDs, particularly their renal, gastrointestinal, and cardiovascular effects, should always be considered, and NSAIDs should be used in the lowest possible dose for the shortest possible duration for all patients.”

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The clinical manifestations of COVID-19 vary widely across different patient populations, despite the virus’s limited genetic diversity. To better understand the mechanisms that drive the diverse responses of infected individuals (hosts) among different patient demographics, researchers extracted and sequenced viral RNA from swabs collected from 430 COVID-19 positive cases and 54 negative controls. The scientists then analyzed the hosts’ antiviral and immune responses across infection status, viral load, age and sex.

Researchers found that immune cell responses were not activated until after three days following the onset of infection, and that immune cell composition and function fluctuated with viral loads, in a way that suggested a dysfunctional antiviral response in males and the elderly. While these findings have important implications for development of immunomodulatory treatments for SARS-CoV-2, additional studies are needed as swabs were taken from the nasopharynx, which is not a sensitive anatomic location for accurately examining markers of systemic inflammation.

Elderly individuals as well as men have suffered higher mortality rates since the onset of the SARS-CoV-2 pandemic and may indicate weaker immune responses underlying poor clinical outcomes. According to the authors, “Collectively, our data demonstrate that host responses to SARS-CoV-2 are dependent on viral load and infection time with observed differences due to age and sex that may contribute to disease severity.”

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The frog, Megaelosia bocainensis, was among seven total species — including four other declining species, and two that had disappeared locally for many years — that were detected. The findings appeared in a paper, “Lost and Found: Frogs in a Biodiversity Hotspot Rediscovered with Environmental DNA,” published in August in Molecular Ecology.

Megaelosia bocainensis. A disappeared species from Parque Nacional da Serra da Bocaina, Brazil, known only from this museum specimen collected in 1968, and detected by eDNA surveys. In the study, the researchers collected and screened environmental DNA (eDNA) in the biodiverse Atlantic Coastal Forest and Cerrado grasslands of Brazil.

The eDNA technique offers a way to survey that can confirm the presence of species undetected by traditional methods, providing a tool for conservation scientists to evaluate the presence of threatened species, especially those with low population densities and those not seen in years.

After careful research to identify species at various levels of threat in these regions of Brazil, the researchers used the eDNA method to search for 30 target amphibian species in six localities where the frogs were known to previously live.

“Little bits of DNA in the environment don’t tell us about how many individuals there are or whether those individuals are healthy, but it does tell us that the species is still present,” said senior author Kelly Zamudio, the Goldwin Smith Professor of Ecology and Evolutionary Biology in the College of Arts and Sciences.

“This is one more kind of survey data, and for species that are declining or locally disappeared, it not only means they are there, but there’s now the potential to study them in more detail,” she said, noting that for many species, very little is known.

Around the world, conservationists have been challenged to keep pace with declining and disappearing amphibians. At the same time, living organisms leave DNA traces in the soil, water and air. Now, scientists are increasingly using highly sensitive sampling techniques to detect eDNA for conservation purposes.

In the study, the researchers targeted 13 frog species that have totally disappeared and are presumed extinct; 12 frogs that have disappeared locally but are still found in other parts of their range; and five species that were once very abundant and are still there but hard to find.

The researchers hiked into the sampling sites carrying battery packs, a shoebox-sized peristaltic pump and backpacks of sterile tubing. They used the pump and tubing to draw up to 60 liters of stream or pond water through a capsule fitted with a filter for capturing DNA. A buffer was then applied to stabilize and preserve the DNA on the filter.

Back in the lab, the researchers extracted the DNA, genetically sequenced it, weeded out genetic material from humans, pigs, chickens and other organisms until they could isolate all the frog DNA.

“Now you’ve got a subset of genetic sequences that we know only belong to frogs, and then it’s step by step, going finer and finer, until you get to the genus and species you are looking for,” Zamudio said.

Identifying M. bocainensis required clever detective work: The species disappeared long ago, and there were no tissues from which to extract DNA for comparison with the eDNA. But the researchers did have the sequences for all the sister species in the genus Megaelosia and they knew the ranges of the sister species and M. bocainensis.

“We know there’s a Megaelosia there,” Zamudio said, “we just don’t know which one it is, but the only one that has ever been reported there historically is the one that went missing. Do we believe it? That’s how far the analysis can take us.”

Zamudio added that samples from nearby areas may be worth collecting for more signs of M. bocainensis.

Carla Martins Lopes, a researcher at São Paulo State University in Brazil, is the paper’s first author.

The Brazilian National Council for Scientific and Technological Development and the São Paulo Research Foundation funded the study.

“When you’re approaching a big tree with a power struggle from far away, you’ll first hear a lot of acorn woodpeckers calling very distinctly, and see birds flying around like crazy,” says first author Sahas Barve, currently a postdoctoral fellow at the Smithsonian National Museum of Natural History. “When you get closer, you can see that there are a dozen or more coalitions of three or four birds fighting and posturing on branches. One group has to beat all the others to win a spot in the territory, which is really, really rare in animals — even in fantasy novels it usually boils down to one army against the other.”

The chaos of the battles makes studying behavior using direct observation difficult. But Barve and his team had an advantage: they used new radio telemetry technology that allowed them to track the birds’ locations down to the minute. With radio tags, which “sit like a rock-climbing harness with a fanny pack on the woodpecker’s back,” the researchers could learn how much time was spent fighting at the power struggles and where the warriors came from.

Power struggles for co-breeding positions in oak trees with “granaries” — large acorn storage structures built by the birds consisting of acorns stuffed into thousands of individual holes in the bark — involve fighting coalitions formed by groups of non-breeding brothers or sisters from neighboring territories. The radio tag data showed that some birds return day after day and fight for ten hours at a time. “We didn’t think it could be that long because they have to be away from their home territory,” says Barve. “When do they eat? We still don’t know.”

The researchers hypothesized that woodpeckers would fight the hardest for territories closest to their current home, but found that deciding to fight may depend on more complex social cues as they recruit members to join their coalition. “These birds often wait for years, and when there’s the right time and they have the right coalition size, they’ll go and give it their all to win a really good territory,” he says.

The woodpeckers’ complex social behavior also extends to the other group that comes to power struggles: the spectators. “We never really paid attention to them because we were always fixated on the birds that were actually fighting,” Barve says. “We often forget that there are birds sitting on trees watching nearby.” His team found that the biggest battles can attract more than 30 birds, or a third of all woodpeckers in the area, with some traveling more than three kilometers to “come with popcorn and watch the fight for the biggest mansion in the neighborhood.”

The radio tag data also showed that the spectators spend up to an hour a day watching the fights, despite many already having breeding position granaries of their own. For them, the benefits of social information must outweigh the costs of leaving their home territory unattended for considerable amounts of time. Acorn woodpeckers have tight social networks and know everyone’s place due to frequent travels to other territories. “If anything is disruptive to that, or if anything weird happens, they want to go check it out,” he says. “The spectators are probably as interested in the outcome as the fighter is, although the warriors benefit more directly.”

There’s still a lot researchers don’t know about the acorn woodpeckers’ complex social structures, but radio telemetry provides a glimpse into their unique social behaviors. “They potentially have friendships, and they probably have enemies,” Barve says. “With our radio tag data, we can tell when two birds are at the same place at the same time. The next step is to try and understand how their social networks are shaped, and how they vary across the year.”

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