The salaries of the 130 Dutch employees throughout the country were not paid on time in March after the company’s bankruptcy in Belgium resulted in the headquarters being closed within a day. This is because unlike Dutch laws, there is no guarantee that Belgian workers will receive any salary in the case of a bankruptcy, which led to the staff at the headquarters staying home. The salaries for April have not been paid as of yet. The staff of the Dutch stores had requested that the company file for bankruptcy a month ago when their payments were late. They then asked the court to adjourn the hearing until May 6 to help ensure they received payment for their work in April. However, the company filed for bankruptcy at the court in Breda before this date. Louwerier has said that he has no idea who the legal owner is of the items in the Casa stores. “There may be a lien or other reservation on those goods.” That is why he decided to keep the stores closed, saying that it would be “irresponsible” to keep the stores open during this time. The bankruptcy administrator promised to do his best to get the benefits agency UWV to pay the staff salaries for April as quickly as possible. CASA, like many other retail chains, had run into financial difficulties in recent years. The company has previously sought additional financing, closed loss-making stores and reorganised its Belgian head office. © 2012-2025, NL Times, All rights reserved. Volume 15 - 2024 | https://doi.org/10.3389/fmicb.2024.1394661 Parts of this article's content have been modified or rectified in: Erratum: Detection of two alphaviruses: Middelburg virus and Sindbis virus from enzootic amplification cycles in southwestern Uganda Our knowledge of alphavirus genetic diversity is mainly based on viruses isolated from anthropophilic mosquito species Studies on alphaviruses from sylvatic amplification cycles in sub-Saharan Africa have been conducted less often than from epizootic environments To gain insight into alphavirus diversity in enzootic transmission cycles we collected over 23,000 mosquitoes in lowland rainforest and savannah gallery forest in southwestern Uganda and tested them for alphavirus infections We detected Sindbis virus (SINV) in a Culex Culex sp mosquito and Middelburg virus (MIDV) in Eretmapodites intermedius and Mansonia africana MIDV is a mosquito-borne alphavirus that causes febrile illness in sheep and horses and was previously not known to occur in Uganda Full genomes of SINV and MIDV were sequenced showing a nucleotide identity of 99% to related strains Both isolates replicated to high titres in a wide variety of vertebrate cells Our data suggest endemic circulation of SINV and MIDV in Uganda While most studies on SINV have been conducted in Finland knowledge regarding its geographic distribution and incidence in other regions in Africa remains limited Uganda is a hotspot region for arbovirus circulation and emergence, including several arboviruses that have been detected for the first time in Uganda’s forests, such as West Nile virus and Zika virus (Smithburn et al., 1940; Dick et al., 1952). This study aimed to sample mosquitoes from pristine and remote areas in Uganda that have an extraordinary amount of biodiversity (Convention on Biological Diversity, 2016) The objective was to assess the genetic diversity of alphaviruses in mosquitoes within enzootic amplification cycles in Uganda that have so far been neglected Such explorations are crucial for detection and control of circulating (endemic) arboviruses as well as to monitor genetic and phenotypic changes contributing to arbovirus emergence The Uganda Wildlife Authority (UWA) and the Uganda National Council for Science and Technology (UNCST) have approved a research permit for this project (number NS632) Virus growth kinetics were performed in mosquito (C6/36) and human (HEK) cells with a multiplicity of infection (MOI) of 0.1 and 0.01 All experiments were performed in duplicate An aliquot of the cell culture supernatant was taken every 24 h for 4 days to measure the number of viral genome copies by quantitative real-time PCR using the primers and probes MIDV-F (5’-TCAGATTTCACTCCATGCACAATG-3′) MIDV-R (5’-ATGCTCAACATGACTATAGCTAGCA-3′) MIDV-TM (5’-CCGATAAAGGCGGCACAT-3′) SINV-F (5’-TTGAATGTCGTTATCGCCAGC-3′) SINV-R (5’-GTTGTCGTCGCCAATGAACG-3′) and SINV-TM (5’-AGCGGCTTAAAACGTCCAGA-3′) RNA was extracted from infectious cell culture supernatants using TRIzol (Life Technologies) followed by cDNA synthesis and library preparation using the KAPA RNA HyperPrep Kit (Roche) next-generation sequencing was performed using the MiSeq Reagent Kit v3 and a MiSeq desktop sequencer (Illumina) The generated reads were processed using an in-house pipeline and database The 5′ and 3′ genome termini were amplified using 5’ RACE System for Rapid Amplification of cDNA Ends (ThermoFisher Scientific) and sequenced by Sanger sequencing All phylogenies were computed using an estimated fraction of invariable sites and an estimated Gamma shape parameter The bootstrap analysis was conducted based on 1,000 replicate trees The complete genome sequences of one SINV strain and two MIDV strains have been registered in GenBank under accession numbers OR183436 The morphologically identified mosquito species were confirmed by sequencing a fragment of the cytochrome c oxidase I (COI) gene indicating the presence of MIDV in female Mansonia africana (MP61-UG-2019) and Eretmapodites intermedius mosquitoes (MP1299-UG-2019) with 100 and 99.7% nucleotide identities The COI gene sequence of the mosquito sample infected with SINV (MP762-UG-2019) showed a nucleotide identity of 97.5% to Culex watti No vertebrate DNA was identified in the virus-positive individual mosquitoes by blood meal analysis SINV MP762-UG-2019 showed rapid and successful replication in all tested cell lines reaching a plateau of approximately 109 RNA genome copies/ml after 4 dpi Genome copies of MIDV MP61-UG-2019 reached a plateau in all tested cell lines 3 dpi and ZNR cells were approximately 10–100-fold higher than in Llu-L and Vero E6 cells and approximately 1,000-fold lower in HEK293T cells (A) Growth kinetics of MIDV in mosquito (C6/36) goat (ZN-R) and sheep (Llu-L) cells infected with a multiplicity of infection of (m (B) Growth kinetics of SINV in mosquito (C6/36) chicken (DF-I) and blackbird (TME-R) cells infected with a m Figure 2. Phylogenetic relationship of SINV strain MP762-UG-2019. The Maximum likelihood (ML) phylogenetic tree of SINV-I is based on the concatenated nucleotide ORF sequences. The virus sequenced in this study is shown in red. SINV strains are labeled by GenBank Accession number, strain, source of isolation, country, and year of collection. The clades are named after Ling et al. (2019) Bootstrap support values are represented by gray (70–90%) or black (>90%) circles at the respective nodes Phylogenetic relationship of MIDV strains MP61-UG-2019 and MP1299-UG-2019 The Maximum likelihood (ML) phylogenetic tree of MIDV based on the concatenated nucleotide ORF sequences (A) and the structural polyprotein (B) The viruses sequenced in this study are shown in red MIDV strains are labeled by GenBank Accession number Bootstrap support values are represented by gray (70–90%) or black (> 90%) circles at the respective nodes Arbovirus surveillance in hotspot regions of arbovirus emergence provides information on circulating strains and is important for correct diagnosis and disease control we collected over 23,000 mosquitoes in a lowland tropical rainforest and adjacent rural communities in southwestern Uganda and tested them for infection with alphaviruses We detected SINV and MIDV in individual mosquitoes from two different primary habitat types suggesting endemic circulation of these viruses in Uganda The efficient and rapid growth of the Ugandan MIDV isolate on the tested human and livestock cell lines suggests no overt impairment of the potentially enzootic isolate with respect to host range future investigations aiming to identify MIDV in acute febrile patients and animals as well as testing for the presence of neutralizing antibodies against MIDV in humans and animals would be needed to shed light on the geographic distribution and burden of MIDV in Uganda surveillance for and diagnosis of SINV infection is limited in Uganda and knowledge of human infection rates is unknown This study highlights the importance of arbovirus surveillance in hotspot regions of arbovirus emergence for understanding the genetic diversity of circulating arboviruses and for the implementation of respective diagnostics and disease control We identified the endemic circulation of SINV and MIDV in Uganda as well as provided the first detection of MIDV in the country Both viruses infect humans and MIDV also causes disease in livestock The potential impact of these viruses on human and animal health is currently unknown as no seroprevalence studies have been conducted in the region Clinical data from patients with compatible symptoms are also not available It is likely that MIDV and SINV infections are underdiagnosed the geographic distribution of SINV and MIDV in Uganda would need to be investigated for more thorough risk assessments with respect to human and animal health It may also be the case that both viruses are mainly restricted to the sylvatic environments in Uganda The isolation of SINV and / or MIDV from epizootic amplification cycles in Uganda could add knowledge regarding virus adaption and evolution during emergence processes from sylvatic amplification cycles This study further raises questions regarding the pathogenicity host range and amplification cycles of these viruses The datasets presented in this study can be found in online repositories The names of the repository/repositories and accession number(s) can be found in the article/supplementary material The author(s) declare that financial support was received for the research This work was funded by the German Ministry for Education and Research (BMBF) under grant agreement number 01KI1716 and the German Research Foundation under grant agreement number JU 2857/11-1 We thank the Uganda Wildlife Authority (UWA) for granting us access to Semuliki National Park (SNP) and Queen Elizabeth National Park (QENP) We further extend our thanks to the park rangers at SNP and QENP for their assistance and use of UWA facilities Special thanks go to Christian Hieke and Verena Heyde for their support in the laboratory and to George Ongodia and Fred Ssenfuka for their introduction to mosquito identification at UVRI The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest The author(s) declared that they were an editorial board member of Frontiers This had no impact on the peer review process and the final decision All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher Sindbis virus as a human pathogen-epidemiology Complete nucleotide sequence of Middelburg virus isolated from the spleen of a horse with severe clinical disease in Zimbabwe Nucleocapsid and glycoprotein organization in an enveloped virus Convention on Biological Diversity (2016) New York: Convention on Biological Diversity Google Scholar Clinical and neuroradiographic manifestations of eastern equine encephalitis A metagenomics-based survey of the virus diversity in mosquito vectors allows the first detection of Sindbis virus in Burkina Faso Edwards FWBM (1941) Google Scholar Ockelbo disease in Sweden: epidemiological and virological data from the 1982 outbreak DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates Detection and genome characterization of Middelburg virus strains isolated from CSF and whole blood samples of humans with neurological manifestations in South Africa Ecologic studies of mosquitoes and birds as hosts of Ockelbo virus in Sweden and isolation of Inkoo and Batai viruses from mosquitoes Seasonality and biting behavior of Mansonia (Diptera Culicidae) in rural settlements near Porto Velho A supplement to the anophelinae of Africa south of the Sahara: Afrotropical region Johannesburg: South African Institute for Medical Research Google Scholar New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0 and accurate algorithm to estimate large phylogenies by maximum likelihood Annual report of the east African Council for Medical Research Google Scholar Discovery of a novel alphavirus related to Eilat virus The subgenus Stegomyia of Aedes in the Afrotropical region with keys to the species (Diptera:Culicidae) Google Scholar Institute of Medicine Forum on Microbial T (2008) The National Academies Collection: Reports funded by National Institutes of Health Global Climate Change and Extreme Weather Events: Understanding the Contributions to Infectious Disease Emergence: Workshop Summary Google Scholar Mosquitoes of southern Africa: Culicinae and Toxorhynchitinae Google Scholar Isolations of Middelburg virus from Aedes (Ochlerotatus) juppi McIntosh (Diptera: Culicidae) suggestive of a reservoir vector Google Scholar Sindbis and West Nile virus infections in the Witwatersrand-Pretoria region Google Scholar Neutralizing antibodies against flaviviruses and Rift Valley fever virus in Ugandan bats MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform Middelburg virus; a hitherto unknown agent isolated from Aedes mosquitoes during an epizootic in sheep in the eastern Cape Province Clinical and laboratory manifestations of Sindbis virus infection: prospective study Introduction and dispersal of Sindbis virus from Central Africa to Europe Mosquito breeding and opposition stimulant in fruit husks Crossref Full Text | Google Scholar Lundström Sindbis virus polyarthritis outbreak signalled by virus prevalence in the mosquito vectors Lundström Phylogeographic structure and evolutionary history of Sindbis virus Discovery of a unique novel clade of mosquito-associated bunyaviruses Infections in sentinel pigeons by Sindbis and West Nile viruses in South Africa with observations on Culex (Culex) univittatus (Diptera: Culicidae) attracted to these birds Epidemics of West Nile and Sindbis viruses in South Africa with Culex (Culex) univittatus Theobald as vector Google Scholar The etiology of epizootic encephalomyelitis of horses in the san Joaquin valley Crossref Full Text | Google Scholar Arboviruses isolated from mosquitoes collected in Uganda Arboviruses and blood meal sources in zoophilic mosquitoes at Human-wildlife interfaces in Kenya a unique alphavirus with host range restricted to insects by RNA replication Occurrence of arthralgia and specific IgM antibodies three to four years after Ockelbo disease Complete genomic sequence of an Australian Sindbis virus isolated 44 years ago reveals unique Indels in the E2 and nsP3 proteins Re-emergence of chikungunya and o’nyong-nyong viruses: evidence for distinct geographical lineages and distant evolutionary relationships Neurological complications in chikungunya fever PubMed Abstract | Google Scholar Ratnasingham, S., and Hebert, P. D. N. (2007). BOLD: The Barcode of Life Data System. Molecular Ecology Notes, pp. pp. 355–364. Available at: http://www.barcodinglife.org Google Scholar A simple method of estimating fifty per cent endpoints12 Crossref Full Text | Google Scholar Barcoding blood meals: new vertebrate-specific primer sets for assigning taxonomic identities to host DNA from mosquito blood meals Sem Ouilibona Full-length genome sequence of a Sindbis virus strain isolated from Culex cinereus in 1977 in bozo Evolutionary analyses of Sindbis virus strains isolated from mosquitoes in Kenya Ockelbo disease: epidemic arthritis-exanthema syndrome in Sweden caused by Sindbis-virus like agent A neurotropic virus isolated from the blood of a native of Uganda 1 Crossref Full Text | Google Scholar Neutralizing antibodies for certain viruses in the sera of human beings residing in northern Natal Ethiopian culicidxe–eretmapodites theobald: description of four new species of the chrysogaster group with notes on the five known species of this group Proceedings of the royal entomological society of london series b Google Scholar Zoonotic alphaviruses in fatal and neurologic infections in wildlife and nonequine domestic animals Sequence coding for the alphavirus nonstructural proteins is interrupted by an opal termination codon PubMed Abstract | Crossref Full Text | Google Scholar Sindbis virus outbreak and evidence for geographical expansion in Finland doi: 10.2807/1560-7917.ES.2022.27.31.2200580 MEGA11: molecular evolutionary genetics analysis version 11 The isolation of Coxsackie-like viruses from mosquitoes PubMed Abstract | Google Scholar Complete genome sequences of two Middelburg viruses isolated from arthropods in the Central African Republic Pogosta disease: clinical observations during an outbreak in the province of North Karelia Van Niekerk Sindbis and Middelburg Old World alphaviruses associated with neurologic disease in horses Global expansion of chikungunya virus: mapping the 64-year history Genetic and phenotypic changes accompanying the emergence of epizootic subtype IC Venezuelan equine encephalitis viruses from an enzootic subtype ID progenitor Arrival of chikungunya virus in the new world: prospects for spread and impact on public health Re-emergence of epidemic Venezuelan equine encephalomyelitis in South America an alphavirus infecting farmed Atlantic Salmon The isolation of SINDBIS virus from the mosquito mansonia (coquillettidia) fuscopennata (theobald) in Uganda PubMed Abstract | Crossref Full Text | Google Scholar Rwego IB and Junglen S (2024) Detection of two alphaviruses: Middelburg virus and Sindbis virus from enzootic amplification cycles in southwestern Uganda Received: 01 March 2024; Accepted: 13 May 2024; Published: 28 May 2024 Copyright © 2024 Graff, Eibner, Ochieng, Jones, Nsubuga, Lutwama, Rwego and Junglen. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) distribution or reproduction in other forums is permitted provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited in accordance with accepted academic practice distribution or reproduction is permitted which does not comply with these terms *Correspondence: Sandra Junglen, c2FuZHJhLmp1bmdsZW5AY2hhcml0ZS5kZQ== Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher. 94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish. Volume 15 - 2024 | https://doi.org/10.3389/fmicb.2024.1469636 Detection of two alphaviruses: Middelburg virus and Sindbis virus from enzootic amplification cycles in southwestern Uganda An Erratum on Detection of two alphaviruses: Middelburg virus and Sindbis virus from enzootic amplification cycles in southwestern Uganda by Graff, S. L., Eibner, G. J., Ochieng, J. R., Jones, T. C., Nsubuga, A. M., Lutwama, J. J., Rwego, I. B., and Junglen, S. (2024). Front. Microbiol. 15:1394661. doi: 10.3389/fmicb.2024.1394661 Due to a production error, there was a mistake in Table 2 as published. In the third column, the aa substitutions in the first three rows are incorrectly formatted and the aa substitutions in the sixth and seventh row are each missing a letter. The corrected Table 2 appears below Non-synonymous substitutions and putative polyprotein position of MP762 to clade B viruses Citation: Frontiers Production Office (2024) Erratum: Detection of two alphaviruses: Middelburg virus and Sindbis virus from enzootic amplification cycles in southwestern Uganda Received: 24 July 2024; Accepted: 24 July 2024; Published: 20 August 2024 Copyright © 2024 Frontiers Production Office. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) *Correspondence: Frontiers Production Office, cHJvZHVjdGlvbi5vZmZpY2VAZnJvbnRpZXJzaW4ub3Jn Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher 94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish AVBOB celebrates a milestone by rewarding its members on a large scale MIDDELBURG - Reinhardt Steyn and his three co-accused appeared in court on Wednesday WATCH: Middelburg farmer allegedly paid for aid in double murder It's believed the farmer accused Thembeni Mnguni and Wanda Mahlangu of stealing sheep Police in Middelburg seized a fake weapon on Wednesday after someone waved it from a moving car It is not clear where exactly the incident took place which bear a European safety certification the toy guns are prohibited from being used to threaten or shoot at people A Middelburg neighborhood officer shared details of the incident on Instagram stating that weapons resembling real firearms can be confiscated and the act is considered a criminal offense The officer noted that the individual who reported the incident was understandably alarmed by the situation The rise in incidents involving gelblasters has been a concern for authorities the police reported an increase in these types of incidents which was partly attributed to a trend on TikTok where videos circulated showing people shooting gelblasters at others A police spokesperson explained that such situations could escalate into dangerous encounters as officers on the scene may not immediately recognize the weapon as a toy While some toys can be immediately identified as such other gelblasters can look rather realistic “It’s certainly possible that you could be held at gunpoint by police it’s crucial that individuals follow instructions carefully,” the spokesperson said Mining and Technical Exhibition (MTE) will drive innovation to the coal face on 14 May 2025.The annual MTE Middelburg/Emalahleni (Witbank) Mining & Industrial Expo will take place in Middelburg this year at Steelcrest High School on 14 May 2025 – rescheduled due to the fact that COALSAFE 2025 takes place on 8 May this year Last year 115 companies booked stands and 400 visitors came through to see innovation in action Historically this is MTE’s main coal exhibition the company has added a general industrial element to the focus on mining houses “Last year 115 companies booked stands and 400 visitors came through to see innovation in action and this year we expect an even greater turnout!” says Andrew Macnamara MTE always customises the expertise exhibiting at the tradeshow to answer the call of the industry operations and commodity focus in the area making sure that the exhibition “speaks” to the challenges of industry in the area through relevant exhibitors Macnamara notes that in his recent conversations with the operations in Middelburg in preparation for the show so its time to concentrate on maintenance and efficiency as producers streamline their operations to remain competitive in the current market environment.” month-on-month coal production contracted (-8.8%)1 Transnet rail and port inefficiencies remain a major constraint despite a notable improvement in the tonnages of coal railed in the latter stages of 2024 Coal railed to the privately owned and operated Richards Bay Coal Terminal (RBCT) “improved to around 52 million tonnes in 2024 up from 47.9 million tonnes in 2023…This is still significantly below the levels of more than 70 million tonnes Transnet railed annually to RBC between 2017 and 2020” 2 Although recent collaborative efforts by the Transnet Freight Rail Industry Recovery Team have provided some relief criminal activity and aging infrastructure remain significant obstacles to achieving efficient rail performance “Coal companies continue to work closely with Transnet to secure the 600km rail line from the coal fields to Richards Bay aiming to mitigate the impact of cable theft and vandalism that act as a constraint to exports.” With an annual output reaching approximately 235 million tonnes in South Africa coal is the country’s most significant mining commodity by production volume and it remains the backbone of South Africa’s energy sector – employing approximately 174 515 people directly in 2024 Minerals Council has noted that the global narrative on coal has been more nuanced after the European energy crisis in 2022 when the continent scrambled to find alternative energy sources after being cut off from Russian gas amid the Ukraine war There is now greater recognition that although the energy transition is an absolute imperative coal will still have a role to play for some time in providing baseload power where it is now considered to be a “critical mineral” because of the important role it plays in sustaining the economy Register for free to gain access the digital library for African Mining publications Click here to send us a message JOHANNESBURG - Middelburg farmer Reinhardt Steyn and his co-accused are also back in court They face charges of kidnapping and assault Murder charges are expected to be added after the bodies of their victims were found The remains were discovered last week after police got a tip-off assaulted and set alight after being accused of stealing sheep JOHANNESBURG - Two bodies burned beyond recognition have been found by police following a tip-off The bodies are believed to be those of Wanda Mahlangu and Thembeni Mnguni They are believed to have been assaulted and killed by farmer Reinhardt Steyn and security guard Dumisani Masilela READ: 2 bodies believed to be Middleburg murder victims found The suspects appeared in court on Thursday for a bail application Mpumalanga's acting Police Commissioner Major General Zeph Mkhwanazi said a lot of processes need to be performed to identify the victims "We will allow the investigation to take place and we hope without doubt more information will come forward," he said.