Biology subjects

Sadiq, S.

Publications and source records attributed to Sadiq, S..

8 recordsLinked to original sources

bioRxiv2026

RNA virus discovery in Australian camelids reveals divergent picornaviruses and the convergent evolution of upstream ORFs

Invasive species can impact viral ecology, evolution and emergence by acquiring and disseminating viruses absent from native hosts. However, the extent to which invasive species harbour previously unrecognized RNA viruses and transmit these to native species is uncertain. We performed metatranscriptomic sequencing of invasive camelids in Australia and identified several previously undescribed vertebrate-associated RNA viruses, including an astrovirus closely related to avian-associated viruses suggesting a recent host jump. We also identified highly divergent picornaviruses that differed sufficiently from recognized taxa in genome organization and polyprotein phylogeny to establish a new genus. Notably, one virus encoded a putative upstream ORF (uORF) in the 5' genomic region. Across the Picornaviridae, putative uORF gain and loss appear to have occurred multiple times independently. In addition, although most of these uORF-encoded proteins exhibited little to no amino acid sequence homology, a subset showed overlapping ranges of secondary structure composition and intrinsic disorder, and when heterologously expressed, these proteins were translated and triggered reproducible transcriptional responses in a cell line. While no single pathway was uniformly affected across all uORFs, distinct uORFs from divergent lineages consistently perturbed overlapping sets of cellular pathways, supporting broadly analogous functional effects despite a lack of sequence homology. These findings demonstrate that uORFs represent a recurrent and selectable functional module within RNA virus genomes, suggest that the upstream genomic position itself constitutes a "hotspot" for the repeated acquisition of a functional module, and provide experimental evidence that their functional properties have converged across evolutionarily independent lineages. Author SummaryRNA viruses evolve under strong genomic constraints, forcing them to repeatedly adopt similar solutions to common challenges posed by their host environment. Invasive species can impact viral ecology, evolution and emergence by acquiring and disseminating non-native viruses. By characterizing RNA viruses infecting invasive camelids in Australia, we discovered previously unrecognized RNA viruses and recurrent patterns of genome evolution. Notably, we identified a small additional gene located at the 5' genomic region, known as an upstream open reading frame (uORF). uORFs were independently acquired and lost across multiple viral lineages, revealing that the same genomic region can be repeatedly exploited to acquire new functions. Although uORF-encoded proteins share little or no amino acid sequence homology, some of the encoded proteins showed comparable secondary structure composition and induce overlapping host cellular responses when expressed in cells. Hence, RNA viruses that have followed different evolutionary paths can converge on similar functional strategies.

microbiology↗

bioRxiv2025

Risk reshapes amygdala representation of choice

Modifying behavior in response to changing environmental conditions is a crucial adaptive function. This capacity is exemplified when animals curtail pursuit of a valued outcome that risks being punished by aversive consequences, but the mediating brain mechanisms remain poorly understood. Here, using in vivo cellular-resolution calcium (Ca2+) imaging, optogenetics and chemogenetics, we show that risk of punishment dramatically alters animals choice between a large/risky and small- /safe reward and produces novel, causally necessary, patterns of activity in basolateral amygdala (BLA) neurons. We find that experience of a punished outcome generates a BLA representation that is selectively replayed when animals subsequently abort choice of the large/risky reward option. Additionally, we show that risk leads to the incorporation of newly encoding BLA neurons into the pre-choice representation, which predicts shifting away from the large/risky option. These findings reveal how dynamic reshaping of BLA representations underpins behavioral flexibility in the face of risk.

neuroscience↗

bioRxiv2025

Limited effect of short- to mid-term storage conditions on an Australian farmland soil RNA virome

Soils represent one of the largest and most diverse reservoirs of microbial life on Earth, yet their associated RNA viruses remain underexplored compared to animal and aquatic systems. Viral discovery in soils has been further limited by technical hurdles, particularly obtaining sufficient yields of high-quality RNA for sequencing. To address this, we evaluated a range of storage and preservation strategies, including the use of commercial preservative solutions and ultra-cold snap-freezing, followed by standardised RNA extraction, sequencing, and virus discovery pipelines. This work aimed to establish minimum sample storage requirements that maintain RNA integrity, generate sufficient RNA sequencing data, and subsequently enable reliable soil virome characterisation. While no preservative solution proved effective, "neat" soil samples were stable at 2-8{degrees}C and -30{degrees}C for at least two weeks, and at -80{degrees}C for at least three months, with no measurable reduction in RNA quality, sequencing data, or viral abundance and diversity. From 32 resulting sequencing libraries, we identified 1,475 putative novel RNA viruses, with the majority belonging to the microbe-associated phylum Lenarviricota. Several novel viruses formed divergent clusters with other environmentally derived sequences distantly related to traditionally animal-associated families such as the Astroviridae and Picornaviridae. Furthermore, unique clusters within the Picobirnaviridae, Alsuvurucetes, Ghabrivirales, and Amabiliviricetes comprised exclusively Australian viruses, suggesting instances of region-specific evolution. Together, these findings highlight soils as rich reservoirs of RNA viral diversity and provide practical minimum standards for storage, expanding opportunities to investigate the ecological and evolutionary roles of RNA viruses in terrestrial systems. ImportanceRNA viruses are the most abundant and diverse biological entities on Earth and are likely present in all other organisms and ecosystems, including soil-dwelling invertebrates, microbes, and plants. Despite this, their diversity and role in soil systems remains largely unknown. Methodological challenges in preserving and extracting sufficient quantities of RNA from soils have hindered the study of these communities. Here, we identified 1,475 previously undescribed RNA viruses in Australian soils while systematically testing different preservation strategies. The significance of our research lies in the demonstration that snap-freezing soil is a viable and robust storage strategy for at least three months, while also highlighting the extraordinary scale of viral diversity present in terrestrial environments. This work establishes a foundation for reliable exploration of terrestrial RNA viruses, improving the accessibility of more remote environmental viromes and enabling future efforts to integrate them into broader models of microbial ecology and ecosystem function.

microbiology↗

bioRxiv2025

Diverse Viral Pathogens in Australian Canines: Limited Geographic Structure and the First Detection of an RNA Virus in Dingoes

Viruses impose a substantial disease burden on dogs and the close relationship between dogs and humans may facilitate zoonotic disease emergence. Australias geographic isolation, strict biosecurity measures and native dingo populations present a unique model for understanding the spread and evolution of canine viruses. However, aside from a few well-characterised pathogens, genomic data are scarce for many common dog viruses, limiting our understanding of their evolution and disease ecology. Using a metatranscriptomic approach we identified the viruses in dogs and dingoes from various geographical locations across mainland Australia and sample types, revealing 86 vertebrate-associated viruses belonging to 16 distinct species, including a new vesivirus-like species. Many of the viruses identified here have not previously been sequenced in Australia. We identified important dog pathogens associated with canine infectious respiratory disease syndrome--such as canine pneumovirus, canine herpesvirus, and canine respiratory coronavirus--and gastroenteritis, including canine parvovirus, canine coronavirus, and rotavirus A. The sequences of Australian canine viruses often occupied multiple distinct clades phylogenetically and had little geographic structure, suggesting multiple virus introductions and subsequent spread across the country. Notably, we identified the first RNA virus - rotavirus A - in a dingo. This virus was phylogenetically distinct from dog-associated rotavirus A sequences and more closely related to viruses found in humans and bats, indicative of the past cross-species transmission of a reassortant virus into dingoes, and shows dingoes and domestic dogs may have distinct viromes. Our findings expand the knowledge of viral diversity in Australian canines, improving our understanding of viral movement into and within Australia, as well as the potential zoonotic risks associated with dogs and dingoes.

microbiology↗

bioRxiv2024

Genomic and phylogenetic features of the Picobirnaviridae suggest microbial rather than animal hosts

The RNA virus family Picobirnaviridae has traditionally been associated with the gastrointestinal systems of terrestrial mammals and birds, with the majority of viruses detected in animal stool samples. Metatranscriptomic studies of vertebrates, invertebrates, microbial communities, and environmental samples have resulted in an enormous expansion of the genomic and phylogenetic diversity of this family. Yet picobirnaviruses remain poorly classified, with only one genus and three species formally ratified by the International Committee of Virus Taxonomy. Additionally, an inability to culture picobirnaviruses in a laboratory setting or isolate them in animal tissue samples, combined with the presence of bacterial genetic motifs in their genomes, suggests these viruses may represent RNA bacteriophage rather than being associated with animal infection. Utilising a data set of 2,286 picobirnaviruses sourced from mammals, birds, reptiles, fish, invertebrates, microbial communities, and environmental samples, we identified seven consistent phylogenetic clusters likely representing picobirnavirus genera that we tentatively name Alpha-, Beta-, Gamma-, Delta-, Epsilon-, Zeta-, and Etapicobirnavirus. A statistical analysis of topological congruence between virus-host phylogenies revealed more frequent cross-species transmission than any other RNA virus family. In addition, bacterial ribosomal binding site motifs were more enriched in picobirnavirus genomes than in the two groups of established RNA bacteriophage - the Leviviricetes and Cystoviridae. Overall, our findings support the hypothesis that the Picobirnaviridae have bacterial hosts and provide a lower-level taxonomic classification for this highly diverse and ubiquitous family of RNA viruses.

microbiology↗

bioRxiv2023

Australian terrestrial environments harbour extensive RNA virus diversity

Australia is home to a diverse range of unique native fauna and flora. To address whether Australian ecosystems also harbour unique viruses, we performed meta-transcriptomic sequencing of 16 farmland and sediment samples taken from the east and west coasts of Australia. We identified 2,562 putatively novel viruses across 15 orders, the vast majority of which belonged to the microbe-associated phylum Lenarviricota. In many orders, the novel viruses identified here comprised entirely new clades, such as the Nodamuvirales and Ghabrivirales. Novel viruses also fell between established genera or families, such as in the Cystoviridae and Picornavirales, while highly divergent lineages were identified in the Martellivirales and Ghabrivirales. Viral abundance and alpha diversity were influenced by sampling site, soil type and land use, but not by depth from the surface. In sum, Australian soils and sediments are home to remarkable viral diversity, reflecting the biodiversity of local fauna and flora.

microbiology↗

bioRxiv2022

RdRp-scan: A Bioinformatic Resource to Identify and Annotate Divergent RNA Viruses in Metagenomic Sequence Data

Despite a rapid expansion in the number of known RNA viruses following the advent of metagenomic sequencing, the identification and annotation of highly divergent RNA viruses remains challenging, particularly from poorly characterized hosts and environmental samples. Protein structures are more conserved than primary sequence data, such that structure-based comparisons provide an opportunity to reveal the viral "dusk matter": viral sequences with low, but detectable, levels of sequence identity to known viruses with available protein structures. Here, we present a new open computational and resource - RdRp-scan - that contains a standardized bioinformatic toolkit to identify and annotate divergent RNA viruses in metagenomic sequence data based on the detection of RNA dependent RNA polymerase (RdRp) sequences. By combining RdRp-specific Hidden Markov models (HMM) and structural comparisons we show that RdRp-scan can efficiently detect RdRp sequences with identity levels as low as 10% to those from known viruses and not identifiable using standard sequence-to-sequence comparisons. In addition, to facilitate the annotation and placement of newly detected and divergent virus-like sequences into the known diversity of RNA viruses, RdRp-scan provides new custom and curated databases of viral RdRp sequences and core motif, as well as pre-built RdRp alignments. In parallel, our analysis of the sequence diversity detected by RdRp-scan revealed that while most of the taxonomically unassigned RdRps fell into pre-established clusters, some sequences cluster into potential new orders of RNA viruses related to the Wolframvirales and Tolivirales. Finally, a survey of the conserved A, B and C RdRp motifs within the RdRp-scan sequence database revealed additional variations of both sequence and position, which might provide new insights into the structure, function and evolution of viral RdRps.

microbiology↗

bioRxiv2018

Evidence for the Modulation of the Immune Response in Peripheral Blood Mononuclear Cells after Stimulation with a High Molecular Weight β-glucan Isolated from Lactobacillus fermentum Lf2

Lactobacillus fermentum Lf 2 produces large amounts of exopolysaccharides under optimized conditions ([~]2 g/L, EPS) which have been shown to possess immunomodulatory activity. In this study, the crude EPS was fractionated to give a high molecular weight (HMw) homoglycan and a mixture of medium molecular weight heteroglycans. The HMw EPS was isolated and identified as a {beta}-glucan.\n\nPeripheral blood mononuclear cells (PBMC) were pre-treated with purified polysaccharide to determine if the HMw {beta}-glucan is responsible for the immunomodulatory activity. Cells were also stimulated with either lipopolysaccharide (LPS) or phytohemagglutinin (PHA) and their effects, both with and without {beta}-glucan pre-treatment, compared.\n\nExposure of the cells to {beta}-glucan increased their metabolic activity and whilst a small but statistically significant drop in CD14 expression was observed at Day 1, the levels were significantly elevated at Day 2. High levels of CD14 expression were observed in cells initially exposed to the {beta}-glucan and subsequently stimulated with either LPS or PHA. In contrast, reduced levels of TLR-2 expression were observed for cells initially exposed to the {beta}-glucan and subsequently stimulated with LPS.\n\nTNF- levels were elevated in {beta}-glucan treated cells (Day1) with the levels dropping back once the {beta}-glucan had been removed (Day 2). The stimulants LPS and PHA both induced significant rises in TNF- levels, however, this induction was completely (LPS) or partially blocked (PHA) in {beta}-glucan pre-treated cells.\n\nThe results indicate a role for the bacterial {beta}-glucan in modulating the immune response following exposure to agonists such as bacterial LPS.

microbiology↗