Original Host Research Articles

SARS-CoV-2 host prediction based on virus-host genetic features

The genetic diversity of the Coronaviruses gives them different biological abilities, such as infect different cells and/or organisms, a wide spectrum of clinical manifestations, their different routes of dispersion, and viral transmission in a specific host. In recent decades, different Coronaviruses have emerged that are highly adapted for humans and causing serious diseases, leaving their host of unknown origin. The viral genome information is particularly important to enable the recognition of patterns linked to their biological characteristics, such as the specificity in the host-parasite relationship. Here, based on a previously computational tool, the Seq2Hosts, we developed a novel approach which uses new variables obtained from the frequency of spike-Coronaviruses codons, the Relative Synonymous Codon Usage (RSCU) to shed new light on the molecular mechanisms involved in the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) host specificity. By using the RSCU obtained from nucleotide sequences before the SARS-CoV-2 pandemic, we assessed the possibility of know the hosts capable to be infected by these new emerging species, which was first identified infecting humans during 2019 in Wuhan, China. According to the model trained and validated using sequences available before the pandemic, bats are the most likely the natural host to the SARS-CoV-2 infection, as previously suggested in other studies that searched for the host viral origin.

In search of COVID-19 transmission through an infected prey.

This paper considers a nonlinear dynamical model of an ecosystem, which has been established through combining the classical Lotka–Volterra model with the classic SIR model. This nonlinear system consists of a generalist predator that subsists on two prey species in which disease is becoming endemic in one of them. The dynamical analysis methods prove that the system has a chaotic attractor and extreme multistability behavior, where there are infinitely many attractors that coexist under certain conditions. The occurrence of extreme multistability demonstrates the high sensitivity of the system for the initial conditions, which means that tiny changes in the original prey species could enlarge and be widespread, and that could confirm through studying the complexity of the time series of the system’s variables. Simulation results of the sample entropy algorithm show that the changes in the system’s variables expand over time. It is reasonable now to consider the endemic in the prey species of the system could evolve to be pandemic such as COVID-19. Consequently, our results could provide a foresight about the unpredictability of the COVID-19 outbreak in its original host species as well as after the transmission to other species such as humans.

Engineering a synthesis-friendly serum responsive promoter for antibiotic selection of genomic integration in cell-based therapy.

For clinical cell-based therapies (e.g. CAR-T cells), the genomic integration of therapeutic genes into cells are selected using inefficient resistance genes of host origin to avoid potential immune response from using more efficient resistance genes of foreign origin. In principle, a serum-responsive promoter could express efficient resistance genes during the cell manufacturing stage that could then diminish during in vivo administration. To avoid genomic instability, we designed a synthesis-friendly serum-responsive promoter (SFSp) with no extreme GC ratios or repeats greater than 9 base pairs. SFSp was used to express a fluorescent reporter, whose expression was diminished after serum starvation. Furthermore, SFSp could be used in replacement of weak promoters (e.g. SV40p) for expressing efficient resistance genes (e.g. blasticidin resistance) from genomic integration via lentiviral infection. Thus, the regulation of resistance genes using SFSp could be a valuable tool in cell-based therapeutics to increase selection efficiency and reduce immunogenicity.

Pharmaceutically Active Microbial AhR Agonists as Innovative Biodrugs in Inflammation.

Alterations of the microbiome occur in inflammatory and autoimmune diseases, a finding consistent with the role of the microbiome in the maintenance of the immune system homeostasis. In this regard, L-tryptophan (Trp) metabolites, of both host and microbial origin, act as important regulators of host–microbial symbiosis by acting as aryl hydrocarbon receptor (AhR) ligands. The intestinal and respiratory barriers are very sensitive to AhR activity, suggesting that AhR modulation could be a therapeutic option to maintain the integrity of the epithelial barrier, which has substantial implications for health even beyond the mucosal site. A number of studies have highlighted the capacity of AhR to respond to indoles and indolyl metabolites, thus positioning AhR as a candidate indole receptor. However, the context-and ligand-dependent activity of AhR requires one to resort to suitable biopharmaceutical formulations to enable site-specific drug delivery in order to achieve therapeutic effectiveness, decrease unwanted toxicities and prevent off-target effects. In this review, we highlight the dual activity of the microbial metabolite indole-3-aldehyde at the host–microbe interface and its ability to orchestrate host pathophysiology and microbial symbiosis and discuss how its proper clinical development may turn into a valuable therapeutic strategy in local and distant inflammatory diseases.

Impacts of Apis cerana gut microbes on Nosema ceranae proliferation in Apis mellifera

The gut microbiota is a complex ecosystem including both beneficial and harmful microbes, which is essential for the metabolism, health and immunity of the host. Fecal microbiota transplantation (FMT) is an effective treatment for diseases of altered intestinal microbiota. Nosema ceranae, a parasitic fungus in intestinal epithelial cells, destroys the honey bee's gut integrity. This study provided the first report that the number of spores was significantly lower in the native host (Asian honey bee) compared with the novel host (European honey bee). The treatment effect of FMT was denoted by feeding the gut tissue solution from the original host to the new host. The results showed that FMT did not significantly decrease the spore load but enhanced the expression levels of immune genes in the Toll pathway. Our data confirmed that the native host inhibits N. ceranae proliferation. Our data suggest microbes of the native host could be an alternative approach to treat the parasite in the novel host.

Systematic Tracing of Susceptible Animals to SARS-CoV-2 by a Bioinformatics Framework.

Since the outbreak of SARS-CoV-2 in 2019, the Chinese horseshoe bats were considered as a potential original host of SARS-CoV-2. In addition, cats, tigers, lions, mints, and ferrets were naturally or experimentally infected with SARS-CoV-2. For the surveillance and control of this highly infectious disease, it is critical to trace susceptible animals and predict the consequence of potential mutations at the binding region of viral spike protein and host ACE2 protein. This study proposed a novel bioinformatics framework to systematically trace susceptible animals to SARS-CoV-2 and predict the binding affinity between susceptible animals’ mutated/un-mutated ACE2 receptors. As a result, we identified a few animals posing a potential risk of infection with SARS-CoV-2 using the docking analysis of ACE2 protein and viral spike protein. The binding affinity of some of these species is weaker than that of humans but more potent than that of Chinese horseshoe bats. We also found that a few point mutations in human ACE2 protein or viral spike protein could significantly enhance their binding affinity, posing an enormous potential threat to public health. The ancestors of the Omicron may evolve rapidly through the accumulation of mutations in infecting the host and jumped into human beings. These findings indicate that if the epidemic expands, there may be a human-animal-human transmission route, which will increase the difficulty of disease prevention and control.

Did Myzus persicae (Sulzer) from potato reared on a novel host for 15 years retain its host-related properties?

Myzus persicae (Sulzer) is an important agricultural pest worldwide causing major economic losses due to its ability to transmit over 100 viruses including Potato virus Y (PVY). Myzus persicae shows considerable variation with respect to performance on its host plants. The objective of this study was to use a survival experiment, behavioural observations, including observations of probing and feeding behaviour obtained using the electrical penetration graph (EPG) technique, and a PVY acquisition experiment to determine whether or not potato was still the more suitable host for M. persicae originating on potato and reared on a novel host, table beet, for over 15 years. In a survival experiment, the pre-reproductive period was significantly longer while adult survival and whole longevity were significantly lower for M. persicae reared on beet fed beet leaves compared to M. persicae reared on potato fed potato leaves. The number of progenies produced and fecundity were both significantly reduced (90 and 85%, respectively) for M. persicae reared on beet fed beet leaves. Ethological observations and EPG assessment of M. persicae behaviour reared on beet placed on beet leaves showed significantly impaired behavioural responses compared to M. persicae reared on potato placed on potato leaves. The rate of PVY acquisition was the same for M. persicae reared on beet and on potato. These results indicate that after 15 years on table beet, M. persicae still performs better on its original host, potato, and appears to be a specialized potato-adapted genotype.

Comparative Transcriptome Investigation of Nosema ceranae Infecting Eastern Honey Bee Workers

Simple SummaryAt present, interaction between Nosema ceranae and Apis cerana is poorly understood, though A. cerana is the original host for N. ceranae. Here, comparative investigation was conducted using transcriptome data from N. ceranae infecting Apis cerana cerana workers at seven days post inoculation (dpi) and 10 dpi (NcT1 and NcT2 groups) as well as N. ceranae spores (NcCK group). There were 1411, 604, and 38 DEGs identified in NcCK vs. NcT1, NcCK vs. NcT2, and NcT1 vs. NcT2 comparison groups. Additionally, 10 upregulated genes and nine downregulated ones were shared by above-mentioned comparison groups. GO classification and KEGG pathway analysis suggested that these DEGs were engaged in a number of key functional terms and pathways such as cell part and glycolysis. Further analysis indicated that most of virulence factor-encoding genes were upregulated, while a few were downregulated during the fungal infection. Findings in this current work provide a basis for clarifying the molecular mechanism udnerlying N. ceranae infection and host-microsporidian interaction during bee nosemosis.Apis cerana is the original host for Nosema ceranae, a widespread fungal parasite resulting in honey bee nosemosis, which leads to severe losses to the apiculture industry throughout the world. However, knowledge of N. ceranae infecting eastern honey bees is extremely limited. Currently, the mechanism underlying N. ceranae infection is still largely unknown. Based on our previously gained high-quality transcriptome datasets derived from N. ceranae spores (NcCK group), N. ceranae infecting Apis cerana cerana workers at seven days post inoculation (dpi) and 10 dpi (NcT1 and NcT2 groups), comparative transcriptomic investigation was conducted in this work, with a focus on virulence factor-associated differentially expressed genes (DEGs). Microscopic observation showed that the midguts of A. c. cerana workers were effectively infected after inoculation with clean spores of N. ceranae. In total, 1411, 604, and 38 DEGs were identified from NcCK vs. NcT1, NcCK vs. NcT2, and NcT1 vs. NcT2 comparison groups. Venn analysis showed that 10 upregulated genes and nine downregulated ones were shared by the aforementioned comparison groups. The GO category indicated that these DEGs were involved in a series of functional terms relevant to biological process, cellular component, and molecular function such as metabolic process, cell part, and catalytic activity. Additionally, KEGG pathway analysis suggested that the DEGs were engaged in an array of pathways of great importance such as metabolic pathway, glycolysis, and the biosynthesis of secondary metabolites. Furthermore, expression clustering analysis demonstrated that the majority of genes encoding virulence factors such as ricin B lectins and polar tube proteins displayed apparent upregulation, whereas a few virulence factor-associated genes such as hexokinase gene and 6-phosphofructokinase gene presented downregulation during the fungal infection. Finally, the expression trend of 14 DEGs was confirmed by RT-qPCR, validating the reliability of our transcriptome datasets. These results together demonstrated that an overall alteration of the transcriptome of N. ceranae occurred during the infection of A. c. cerana workers, and most of the virulence factor-related genes were induced to activation to promote the fungal invasion. Our findings not only lay a foundation for clarifying the molecular mechanism underlying N. ceranae infection of eastern honey bee workers and microsporidian–host interaction.

Multipurpose medical image watermarking for effective security solutions.

Digital medical images contain important information regarding patient’s health and very useful for diagnosis. Even a small change in medical images (especially in the region of interest (ROI)) can mislead the doctors/practitioners for deciding further treatment. Therefore, the protection of the images against intentional/unintentional tampering, forgery, filtering, compression and other common signal processing attacks are mandatory. This manuscript presents a multipurpose medical image watermarking scheme to offer copyright/ownership protection, tamper detection/localization (for ROI (region of interest) and different segments of RONI (region of non-interest)), and self-recovery of the ROI with 100% reversibility. Initially, the recovery information of the host image’s ROI is compressed using LZW (Lempel-Ziv-Welch) algorithm. Afterwards, the robust watermark is embedded into the host image using a transform domain based embedding mechanism. Further, the 256-bit hash keys are generated using SHA-256 algorithm for the ROI and eight RONI regions (i.e. RONI-1 to RONI-8) of the robust watermarked image. The compressed recovery data and hash keys are combined and then embedded into the segmented RONI region of the robust watermarked image using an LSB replacement based fragile watermarking approach. Experimental results show high imperceptibility, high robustness, perfect tamper detection, significant tamper localization, and perfect recovery of the ROI (100% reversibility). The scheme doesn’t need original host or watermark information for the extraction process due to the blind nature. The relative analysis demonstrates the superiority of the proposed scheme over existing schemes.

Proteolytic Landscapes in Gastric Pathology and Cancerogenesis

Gastric cancer is a leading cause of cancer-related death, and a large proportion of cases are inseparably linked to infections with the bacterial pathogen and type I carcinogen Helicobacter pylori. The development of gastric cancer follows a cascade of transformative tissue events in an inflammatory environment. Proteases of host origin as well as H. pylori-derived proteases contribute to disease progression at every stage, from chronic gastritis to gastric cancer. In the present article, we discuss the importance of (metallo-)proteases in colonization, epithelial inflammation, and barrier disruption in tissue transformation, deregulation of cell proliferation and cell death, as well as tumor metastasis and neoangiogenesis. Proteases of the matrix metalloproteinase (MMP) and a disintegrin and metalloproteinase domain-containing protein (ADAM) families, caspases, calpain, and the H. pylori proteases HtrA, Hp1012, and Hp0169 cleave substrates including extracellular matrix molecules, chemokines, and cytokines, as well as their cognate receptors, and thus shape the pathogenic microenvironment. This review aims to summarize the current understanding of how proteases contribute to disease progression in the gastric compartment.

Prevalence, antimicrobial resistance and virulence genes of Salmonella serovars isolated from humans and animals.

We investigated the prevalence, antimicrobial susceptibility, antimicrobial resistance and virulence genes of Salmonella isolates recovered from humans and different species of animals. Out of 1231 samples, 88 (7.15%) Salmonella isolates were obtained, among which 21 (23.86%) belonged to Salmonella enterica subsp. enterica sero var. Weltevreden, 22 (25%) to S. Enteritidis, 16 (18.2%) to S. Typhi and 14 (15.9%) to S. Newport; 7 (7.95%) isolates were untypable. Among the 88 isolates, 65.90% showed resistance to gentamicin, 61.36% to tetracycline, 61.18% to cefotaxime, 48.86% to trimethoprim, 45.45% to ampicillin, 11.36% to ceftriaxone, 10.22% to chloramphenicol and 7.95% each to ciprofloxacin and cefepime. Most of the isolates were susceptible, with a low MIC (≤ 0.25μg/ml) value, to cefepime, cefotaxime, ciprofloxacin, ceftriaxone and co-trimoxazole and with a moderate MIC (0.5-4μg/ml) to ampicillin, tetracycline, gentamicin and chloramphenicol. The resistance genes blaTEM, tetA and dfrA12 were most prevalent, irrespective of the host of origin of the isolates. While invA was used for molecular detection of Salmonella, other virulence genes, viz. sipA, sipB, sipC, stn and pagN, were also detected in all Salmonella isolates. A total of 38.64% isolates were multidrug-resistant (MDR), and various virulence genes were present among the isolated serovars. This study highlights the importance of continuous monitoring and surveillance for pathogenic Salmonella and their potential risks to both humans and animals.

Full Genome Nobecovirus Sequences From Malagasy Fruit Bats Define a Unique Evolutionary History for This Coronavirus Clade.

Bats are natural reservoirs for both Alpha- and Betacoronaviruses and the hypothesized original hosts of five of seven known zoonotic coronaviruses. To date, the vast majority of bat coronavirus research has been concentrated in Asia, though coronaviruses are globally distributed; indeed, SARS-CoV and SARS-CoV-2-related Betacoronaviruses in the subgenus Sarbecovirus have been identified circulating in Rhinolophid bats in both Africa and Europe, despite the relative dearth of surveillance in these regions. As part of a long-term study examining the dynamics of potentially zoonotic viruses in three species of endemic Madagascar fruit bat (Pteropus rufus, Eidolon dupreanum, Rousettus madagascariensis), we carried out metagenomic Next Generation Sequencing (mNGS) on urine, throat, and fecal samples obtained from wild-caught individuals. We report detection of RNA derived from Betacoronavirus subgenus Nobecovirus in fecal samples from all three species and describe full genome sequences of novel Nobecoviruses in P. rufus and R. madagascariensis. Phylogenetic analysis indicates the existence of five distinct Nobecovirus clades, one of which is defined by the highly divergent ancestral sequence reported here from P. rufus bats. Madagascar Nobecoviruses derived from P. rufus and R. madagascariensis demonstrate, respectively, Asian and African phylogeographic origins, mirroring those of their fruit bat hosts. Bootscan recombination analysis indicates significant selection has taken place in the spike, nucleocapsid, and NS7 accessory protein regions of the genome for viruses derived from both bat hosts. Madagascar offers a unique phylogeographic nexus of bats and viruses with both Asian and African phylogeographic origins, providing opportunities for unprecedented mixing of viral groups and, potentially, recombination. As fruit bats are handled and consumed widely across Madagascar for subsistence, understanding the landscape of potentially zoonotic coronavirus circulation is essential for mitigation of future zoonotic threats.

Genomic Comparisons and Phenotypic Diversity of Dickeya zeae Strains Causing Bacterial Soft Rot of Banana in China.

Bacterial soft rot of banana, caused by Dickeya zeae, is spreading rapidly in important banana growing areas in China and seriously threatens banana production. In this study, we sequenced the high-quality complete genomes of three typical banana strains, MS1 (size: 4,831,702-bp; genome coverages: 538x), MS_2014 (size: 4,740,000-bp; genome coverages: 586x) and MS_2018 (size: 4,787,201-bp; genome coverages: 583x), isolated in 2009, 2014, and 2018, respectively. To determine their genomic and phenotypic diversity with respect to their hosts of origin, they were compared with other D. zeae strains, including another representative banana strain MS2 from China. The sequenced strains were similar in utilization of carbon source and chemical substrates, and general genomic features of GC content, and tRNA and rRNA regions. They were also conserved in most virulence determinants, including gene-encoding secretion systems, plant cell wall degrading enzymes, and exopolysaccharides. We further explored their genomic diversity in the predicted genomic islands (GIs). These GIs were rich in integrases and transposases, where some genomic dissimilarity was observed in the flagellar gene cluster and several secondary metabolite gene clusters. Different constituents of core biosynthetic modules were found within the bacteriocin and aryl polyene (APE) pigment gene clusters, and the strains from banana showed different phenotypes with respect to antibiosis effects and colony pigmentation. Additionally, clustered regularly interspaced short palindromic repeat (CRISPR) and prophage elements, such as type I-F and III-A CRISPR arrays and an intact prophage of MS1-P5, contributed to bacterial diversity. Phylogenetic tree analysis and genome-genome nucleotide comparison confirmed the genomic divergence among the strains isolated from banana. Considering these characteristics, MS2 and MS_2014 probably diverged later than MS1, while MS_2018 was different and more similar to foreign strains isolated from other hosts in several characteristics. Strain MS_2018 caused severe symptoms on banana varieties previously considered moderately resistant or moderately susceptible, including varieties of Cavendish (Musa AAA) and Plantain (Musa ABB). Our study of genomic and phenotypic diversity raises public attention to the risk of spreading new pathogenic variants within banana growing regions and supports development of predictive strategies for disease control.

Molecular detection of Trypanosoma (Trypanosomatidae) in bats from Thailand, with their phylogenetic relationships.

The vast majority of trypanosome species is vector-borne parasites, with some of them being medically and veterinary important (such as Trypanosoma cruzi and Trypanosoma brucei) and capable of causing serious illness in vertebrate hosts. The discovery of trypanosomes in bats emphasizes the importance of bats as an important reservoir. Interestingly, there is a hypothesis that bats are ancestral hosts of T. cruzi. Trypanosome diversity has never been investigated in bats in Thailand, despite being in a biodiversity hot spot. To gain a better understanding of the diversity and evolutionary relationship of trypanosomes, polymerase chain reaction-based surveys were carried out from 2018 to 2020 in 17 sites. A total of 576 bats were captured, representing 23 species. A total of 38 (6.6%) positive samples was detected in ten bat species. Trypanosoma dionisii and Trypanosoma noyesi were identified from Myotis siligorensis and Megaderma spasma, respectively. The remaining 18S rRNA sequences of trypanosomes were related to other trypanosomes previously reported elsewhere. The sequences in the current study showed nucleotide identity as low as 90.74% compared to those of trypanosomes in the GenBank database, indicating the possibility of new species. All bat trypanosomes identified in the current study fall within the T. cruzi clade. The current study adds to evidence linking T. noyesi to a bat trypanosome and further supports the bat host origin of the T. cruzi clade. To the best of authors' knowledge, this is the first study on bat trypanosomes in Thailand and their phylogenetic relationships with global isolates.

Aggressiveness and Mycotoxin Production by Fusarium meridionale Compared with F. graminearum on Maize Ears and Stalks in the Field.

Fusarium meridionale and F. graminearum both cause Gibberella ear rot (GER) and Gibberella stalk rot (GSR) of maize in Brazil, but the former is much more common. Recent work with two isolates of each from maize suggested this dominance could be caused by greater aggressiveness and competitiveness of F. meridionale on maize. We evaluated pathogenicity and toxigenicity of 16 isolates of F. graminearum and 24 isolates of F. meridionale recovered from both wheat and maize. Strains were individually inoculated into ears of four maize hybrids in field trials. GER severity varied significantly between isolates within each species. Although ranges overlapped, the average GER severity induced by F. meridionale (25.2%) was two times as high overall as that induced by F. graminearum (12.8%) for isolates obtained from maize but was similar for those isolated from wheat (19.9 and 21.4%, respectively). In contrast, severity of GSR was slightly higher for F. graminearum (22.2%) than for F. meridionale (19.8%), with no effect of the host of origin. Deoxynivalenol and its acetylated form 15ADON were the main mycotoxins produced by F. graminearum (7/16 strains), and nivalenol toxin was produced by F. meridionale (17/24 strains). Six isolates of F. graminearum and three of F. meridionale also produced zearalenone. Results confirmed that F. meridionale from maize is, on average, more aggressive on maize but also suggested greater complexity related to diversity among the isolates within each species and their interactions with different hybrids. Further studies involving other components of the disease cycle are needed to more fully explain observed patterns of host dominance.

Impacts of Continued Exposure to a Susceptible Host Genotype on Aggressiveness of Phytophthora nicotianae Isolates Adapted to Multiple Sources of Partial Resistance.

Pathogen adaptation can threaten the durability of partial resistance. Mixed plantings of susceptible and partially resistant varieties may prolong the effectiveness of partial resistance, but little is known about how continued exposure to a susceptible genotype can change the aggressiveness of pathogen isolates adapted to a source of partial resistance. The objective of this study was to examine the effects of continued exposure to a highly susceptible tobacco genotype on isolates of Phytophthora nicotianae that had been adapted to partial resistance. Isolates of P. nicotianae previously adapted to two sources of partial resistance were continually exposed to either the original host of adaptation or a susceptible host. After six generations of host exposure, isolates obtained from the partially resistant and the susceptible hosts were compared for their aggressiveness on the resistant host and for differences in expression of genes associated with pathogenicity and aggressiveness. Results suggested that exposure to the susceptible tobacco genotype reduced aggressiveness of isolates adapted to partial resistance in K 326 Wz/- but not of isolates adapted to partial resistance in Fla 301. Quantification of pathogenicity-associated gene expression using qRT-PCR suggested the rapid change in aggressiveness of isolates adapted to Wz-sourced partial resistance may have resulted from modification in gene expression in multiple genes.

Accessory phase perspectives for ore-forming processes and magmatic sulphide exploration in the Labrador Trough, northern Quebec, Canada

The compositions of resistant indicator minerals are diagnostic of their original host environment. They may be used to fingerprint different types of mineral deposit as well as vector towards them. We have characterised the composition of apatite and Fe–Ti oxides in variably mineralised mafic–ultramafic rock units of the Montagnais Sill Complex in the Labrador Trough to assess their suitability for vectoring towards magmatic sulphide occurrences. Two broad types of apatite were identified: (i) fluoro- to hydroxy-apatite (Cl/(Cl+F) < 0.2); and (ii) chloro- to hydroxy-apatite (Cl/(Cl+F) > 0.5). The former reflects variable degrees of degassing and Cl loss during Rayleigh fractionation and is not indicative of Ni–Cu mineralisation or host rock. The latter exists only in sulphidic olivine cumulate units and thus may be used to vector towards similar rock types in the Labrador Trough. Ilmenite is the dominant oxide, except for the upper parts of differentiated gabbroic sills in which titanomagnetite is dominant. Magnetite occurs only as a secondary phase in serpentinised olivine cumulates and is not discriminative for magmatic sulphides. Ilmenite and titanomagnetite in the sulphidic olivine-bearing units have characteristically high Mg (∼1000–10 000 ppm), Cr (∼100–1000 ppm), and Ni (∼10–1000 ppm) concentrations relative to those from other rock units. Their composition is consistent with Fe–Ti oxides derived from evolved sulphide melts in ultramafic-hosted Ni–Cu–(PGE) sulphide deposits and thus may be used to vector towards similar magmatic sulphide occurrences in the Labrador Trough.

P004 Microbiota, not host origin drives ex vivo epithelial response in ulcerative colitis patients and non-IBD controls

Abstract Background Host-microbial interactions in inflammatory bowel disease (IBD) are poorly understood. Furthermore, patients with IBD have microbial dysbiosis and their epithelial cells exhibit intrinsic defects. We aimed to unravel the effect of exposure to microbiota on epithelial cells from both patients with ulcerative colitis (UC) and non-IBD controls. Methods Confluent Transwell® organoid-derived monolayers of 8 UC patients and 8 non-IBD controls were co-cultured for 6 hours with microbiota (3.108 cells) derived from active UC patients (n=3, endoscopic Mayo score ≥2) or healthy volunteer (HV, n=1, selected on high microbial cell count and presence of selected phyla1). For this purpose, fresh faecal samples were filtered and frozen in 0.9% NaCl. Inflammation was re-induced with 100 ng/ml TNF-α, 20 ng/ml IL-1β, and 1 µg/ml flagellin in confluent Transwell® cultures, 24 hours prior to microbiota co-culture. Transepithelial electrical resistance (TEER), 4 kDa fluorescein isothiocyanate (FITC) dextran (2 mg/ml) measurements, and RNA sequencing by Truseq were performed on epithelial cells, and 16S rRNA sequencing on microbiota samples before and after co-culture. Results The transcriptomic response of epithelial cells to microbiota was clearly influenced by the type of microbiota stimulation (no, UC or HV microbiota). However, this response was not influenced by the origin of the epithelial cells (UC or non-IBD controls) (Figure 1A). TEER and FITC dextran measurements showed a strong decrease in epithelial integrity (in both UC and non-IBD epithelium) following stimulation with UC microbiota, in contrast to HV microbiota (Figure 1B). In UC epithelial cells, stimulation with microbiota from UC patients induced a stress induced phenotype including activation of EGR1 signalling, AP-1 family, FOSL genes, MAPK and JNK signalling (Figure 2). Activation of stress pathways (MAPK family cascades) and key markers (e.g. FOSB, FOSL1, MYC, EGR1, ATF3, NOTCH1) was lower after HV microbiota stimulation (Figure 3–4). Expression levels of key markers were UC microbiota specific and similar in epithelial cells of UC and non-IBD controls (Figure 4). Conclusion Not the host (UC vs non-IBD epithelial cells) but the microbial donor (UC vs HV) is driving the transcriptomic response. Epithelial cells from UC patients did not show an increased sensitivity towards microbiota stimulation compared to non-IBD epithelial cells. In contrast, exposure of epithelial cells to UC microbiota was sufficient to induce a strong stress response and barrier disruption. Further research on therapies to restore the microbial balance, to remove the constant trigger of dysbiosed microbiota, is required.

Insight into the symbiotic lifestyle of DPANN archaea revealed by cultivation and genome analyses

Decades of culture-independent analyses have resulted in proposals of many tentative archaeal phyla with no cultivable representative. Members of DPANN (an acronym of the names of the first included phyla Diapherotrites, Parvarchaeota, Aenigmarchaeota, Nanohaloarchaeota, and Nanoarchaeota), an archaeal superphylum composed of at least 10 of these tentative phyla, are generally considered obligate symbionts dependent on other microorganisms. While many draft/complete genome sequences of DPANN archaea are available and their biological functions have been considerably predicted, only a few examples of their successful laboratory cultivation have been reported, limiting our knowledge of their symbiotic lifestyles. Here, we investigated physiology, morphology, and host specificity of an archaeon of the phylum "Candidatus Micrarchaeota" (ARM-1) belonging to the DPANN superphylum by cultivation. We constructed a stable coculture system composed of ARM-1 and its original host Metallosphaera sp. AS-7 belonging to the order Sulfolobales Further host-switching experiments confirmed that ARM-1 grew on five different archaeal species from three genera-Metallosphaera, Acidianus, and Saccharolobus-originating from geologically distinct hot, acidic environments. The results suggested the existence of DPANN archaea that can grow by relying on a range of hosts. Genomic analyses showed inferred metabolic capabilities, common/unique genetic contents of ARM-1 among cultivated micrarchaeal representatives, and the possibility of horizontal gene transfer between ARM-1 and members of the order Sulfolobales Our report sheds light on the symbiotic lifestyles of DPANN archaea and will contribute to the elucidation of their biological/ecological functions.

The First Report of Elaphostrongylus cervi Infection in Two Imported Wapitis (Cervus canadensis) in Slovenia

This study describes two female wapitis (Cervus canadensis) with neurological signs associated with an Elaphostrongylus cervi (E. cervi) infection. The original host of the nematode parasite is the Eurasian red deer (Cervus elaphus), although other cervids and small ruminants may also be affected. The two wapitis imported from Canada were kept in an enclosure with the Slovenian red deer herd. After developing debilitating neurological signs, the wapitis were euthanized and examined for possible causes. A histopathological examination of the brain of the first wapiti revealed severe diffuse perivascular meningoencephalitis with chronic vasculitis, and some cross-sections of nematodes were found in the leptomeninges. A necropsy of the second wapiti revealed severe pachymeningitis and leptomeningitis, where several adult nematode parasites were found. E. cervi was confirmed by molecular methods. The prevalence of E. cervi in the European red deer population is high, but no study has been conducted to assess its prevalence in Slovenia. This was the first confirmation of E. cervi in Slovenia and the first infection with this parasite described in Europe in a wapiti. Elaphostrongylus cervi should also be considered as a differential diagnosis in Europe for all ruminants grazing on pastures frequented by red deer and showing neurological clinical signs.