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van der Linden, A.

Publications and source records attributed to van der Linden, A..

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Shared viral burdens: Evidence of active Usutu virus circulation and multi-arbovirus exposure in migrant and resident birds at wintering locations in Nigeria

BackgroundWest Nile (WNV), Usutu (USUV), and Sindbis (SINV) virus were initially detected in the African region, and subsequently across temperate regions where they were absent. Wild birds are primary reservoirs for these arboviruses and are considered major contributors to their global spread through seasonal migration. To understand the transmission dynamics of arboviruses in wild birds and the potential of migratory birds to spread the viruses at an intercontinental scale, we investigated arboviral infections and exposures in African resident and Palearctic migratory birds at wintering locations in Nigeria. Methodology/Principal FindingsOropharyngeal- and cloacal swabs, feathers and blood were collected from resident and migratory birds at two wintering locations (Amurum and Ngel-Nyaki Forest Reserves). Swabs and feathers were tested using RT-PCR for WNV, USUV and SINV, and blood with ELISA and FRNT90 or PRNT80 for antibodies. 573 birds were sampled between 2021 to 2024 across months coinciding with arrival and departure of migratory birds. USUV RNA was detected in 2.6% of feathers including a positive Icterine warbler and a garden warbler sampled prior to spring migration. None of the swabs was positive for viral RNA but neutralizing antibodies to WNV and USUV were detected in 4.5% of birds. SINV antibodies were also found in 34.1% of birds sampled across the wintering locations. Conclusions/SignificanceOur findings showed that migratory birds can become infected with USUV, and potentially with WNV and SINV during their overwintering periods in Africa and highlighted a wider arbovirus risk in Nigeria. In addition, detections of viral RNA in feathers, but not swabs, suggest feathers may be a suitable matrix for surveillance in the absence of a reliable cold chain. The overall detections in wild birds at these locations highlight the need for further surveillance to define the epidemiology and public health risks of these arboviruses in the region. SynopsisVector-borne viruses like West Nile, Usutu, and Sindbis, were once limited to parts of Africa, but now appear in Europe and other temperate regions. Migratory birds are believed to help spread these viruses across continents. To understand the role of birds in this, we tested resident and migratory birds in Nigeria for evidence of infection. Between 2021 and 2024, we sampled over 570 birds during their winter stay. We found genetic material of Usutu virus in feathers of six birds including two migratory birds just before their return to Europe. While swab tests did not detect active infections, antibodies to these viruses, which indicate past exposure, were found in several birds, especially for Sindbis virus. Our findings show that migratory birds can pick up and possibly carry these viruses across continents. We also found that feathers may be a practical tool for virus detection in areas without reliable refrigeration, supporting easier and broader surveillance of these viruses in Africa and possibly beyond.

microbiology↗

Weight cycling-induced hypothalamic and metabolic tissue immune remodeling is uncoupled from metabolic dysfunctions

BackgroundObesity-induced insulin resistance is associated with white adipose tissue (WAT) and liver inflammation, which are both mitigated by weight loss. However, most individuals undergoing weight loss will regain lost weight, resulting in weight cycling (WC), which may exacerbate metabolic dysfunctions. Here, we studied the immunometabolic impact of WC in mice. MethodsC57BL/6J mice were exposed to two cycles of weight gain and weight loss by alternating between low (LFD) and high-fat diet (HFD) feeding. Animals were sacrificed when WC mice were weight stable for 10 weeks upon weight loss (WC-lean) and after a subsequent exposure to weight regain for 10 weeks (WC-obese), and compared to mice persistently fed LFD (LFD-lean) or HFD (HFD-obese). ResultsBody weight stabilized at a higher level in WC-lean mice after two weight gain/loss cycles compared to LFD-lean controls. While insulin resistance, metabolic tissue inflammation, and hepatic steatosis normalized between the two groups of lean mice, WC-lean mice exhibited features of WAT dysfunction. In the hypothalamus, inflammatory microglia were less abundant in WC-lean mice compared to LFD-lean mice, but mean individual microglial cell volume was larger. WC-obese mice stabilized at slightly lower weight compared with HFD-obese controls. Intriguingly, WC-obese mice exhibited increased WAT macrophages and reduced WAT and liver effector T cells compared to HFD-obese mice, whereas energy intake, body composition, whole-body insulin resistance and hepatic steatosis were similar. ConclusionsOur results suggest that WC in mice differently impacts animals in the weight-stable lean and obese states. WC-lean mice display features of a novel body weight settling point, associated with hypothalamic inflammatory changes. However, metabolic dysfunctions were uncoupled from WC-induced metabolic tissue inflammation in WC-obese mice.

immunology↗

Emergence and Dynamics of Usutu and West Nile Viruses in the Netherlands, 2016-2022

BackgroundMosquito-borne arboviruses, including Usutu virus (USUV) and West Nile virus (WNV), are emerging threats in Europe, with changes in climate, land use, and increased travel and trade influencing their dynamics. Understanding the emergence and establishment of these viruses in new regions is critical for informing targeted mitigation of drivers of emergence and enhancing public and wildlife health preparedness. MethodsSeven years (2016-2022) of interconnected studies were conducted in the Netherlands. Live birds were sampled by volunteer bird ringers, dead birds were referred for sampling by citizen-scientists and zoological institutions, and mosquitoes were trapped. Samples were tested for USUV and WNV using RT-PCR and screened for Orthoflavivirus antibodies using protein microarray and neutralization assays. Sequencing and phylogenetic analyses were performed. FindingsUSUV was first detected in the Netherlands in 2016, caused large outbreaks in birds until 2018, and resurged in 2022. One dominant, enzootic lineage, co-circulated alongside several limited introductions of a second lineage. A localized WNV lineage 2 outbreak occurred in live birds and mosquitoes in 2020, with another positive bird in 2022 and serological evidence of continued circulation. InterpretationWe provide the first comprehensive, multi-year documentation of two emerging arboviruses in the Netherlands. USUV was enzootically maintained over seven years and was associated with substantial bird mortality. WNV is in an early stage of establishment with no bird mortality observed. Our integrated wildlife sampling was crucial in detecting a human WNV outbreak, bringing this emerging threat to public health attention. FundingZonMw, Dutch Research Council (NWO), European Union, Government of the Netherlands

microbiology↗

High number of HPAI H5 Virus Infections and Antibodies in Wild Carnivores in the Netherlands, 2020-2022

In October 2020, a new lineage of clade 2.3.4.4b HPAI virus of the H5 subtype emerged in Europe, resulting in the largest global outbreak of HPAI to date, with unprecedented mortality in wild birds and poultry. The virus appears to have become enzootic in birds, continuously yielding novel HPAI virus variants. The recently increased abundance of infected birds worldwide increases the probability of bird-mammal contact, particularly in wild carnivores. Here, we performed molecular and serological screening of over 500 dead wild carnivores for H5 HPAI virus infection and sequencing of positive materials. We show virological evidence for HPAI H5 virus infection in 0.8%, 1.4% and 9.9% of animals tested in 2020, 2021 and 2022 respectively, with the highest proportion of positives in foxes, polecats and stone martens. We obtained near full genome sequences for seven viruses and detected PB2 amino acid substitutions known to play a role in mammalian adaptation in three of these. Infections were also found in animals without associated neurological signs or mortality. Serological evidence for infection was detected in 20% of the study population. These findings suggest that a higher number of wild carnivores are infected but undetected in current surveillance programs. We recommend increased surveillance in susceptible mammals, irrespective of the presence of neurological signs or encephalitis.

microbiology↗

Rapid SARS-CoV-2 whole genome sequencing for informed public health decision making in the Netherlands

SARS-CoV-2 is a novel coronavirus that has rapidly spread across the globe. In the Netherlands, the first case of SARS-CoV-2 has been notified on the 27th of February. Here, we describe the first three weeks of the SARS-CoV-2 outbreak in the Netherlands, which started with several different introductory events from Italy, Austria, Germany and France followed by local amplification in, and later also, outside the South of the Netherlands. The timely generation of whole genome sequences combined with epidemiological investigations facilitated early decision making in an attempt to control local transmission of SARS-CoV-2 in the Netherlands.

genomics↗

Distributed DNA-based Communication in Populations of Synthetic Protocells

Developing distributed communication platforms based on orthogonal molecular communication channels is a crucial step towards engineering artificial multicellular systems. Here, we present a general and scalable platform entitled Biomolecular Implementation of Protocellular Communication (BIO-PC) to engineer distributed multichannel molecular communication between populations of non-lipid semipermeable microcapsules. Our method leverages the modularity and scalability of enzyme-free DNA strand-displacement circuits to develop protocellular consortia that can sense, process and respond to DNA-based messages. We engineer a rich variety of biochemical communication devices capable of cascaded amplification, bidirectional communication and distributed computational operations. Encapsulating DNA strand-displacement circuits further allows their use in concentrated serum where non-compartmentalized DNA circuits cannot operate. BIO-PC enables reliable execution of distributed DNA-based molecular programs in biologically relevant environments and opens new directions in DNA computing and minimal cell technology.

synthetic biology↗

Global accumulation of circRNAs during aging in Caenorhabditis elegans

Circular RNAs (CircRNAs) are a newly appreciated class of RNAs that lack free 5{acute} and 3{acute} ends, are expressed by the thousands in diverse forms of life, and are mostly of enigmatic function. Ostensibly due to their resistance to exonucleases, circRNAs are known to be exceptionally stable. Here, we examined the global profile of circRNAs in C. elegans during aging by performing ribo-depleted total RNA-seq from the fourth larval stage (L4) through 10-day old adults. Using stringent bioinformatic criteria and experimental validation, we annotated 1,166 circRNAs, including 575 newly discovered circRNAs. These circRNAs were derived from 797 genes with diverse functions, including genes involved in the determination of lifespan. A massive accumulation of circRNAs during aging was uncovered. Many hundreds of circRNAs were significantly increased among the aging time-points and increases of select circRNAs by over 40-fold during aging were quantified by qRT-PCR. The age-accumulation of circRNAs was not accompanied by increased expression of linear RNAs from the same host genes. We attribute the global scale of circRNA age-accumulation to the high composition of postmitotic cells in adult C. elegans, coupled with the high resistance of circRNAs to decay. These findings suggest that the exceptional stability of circRNAs might explain age-accumulation trends observed from neural tissues of other organisms, which also have a high composition of post-mitotic cells. Given the suitability of C. elegans for aging research, it is now poised as an excellent model system to determine if there are functional consequences of circRNA accumulation during aging.

genomics↗