Narrow Host Research Articles

“Sichuanvirus”, a novel bacteriophage viral genus, able to lyse carbapenem-resistant Klebsiella pneumoniae

BackgroundCarbapenem-resistant Klebsiella pneumoniae (CRKP) is a severe threat for human health and urgently needs new therapeutic approaches. Lytic bacteriophages (phages) are promising clinically viable therapeutic options against CRKP. We attempted to isolate lytic phages against CRKP of sequence type 11 and capsular type 64 (ST11-KL64), the predominant type in China.ResultsWe recovered a lytic phage from sewage collected at a wastewater treatment station in Sichuan province, China. We obtained the genome of this phage and found that it is distinct from all known phages with the highest overall DNA similarity (12.5%, 16% coverage and 78.4% identity) with phage vB_EcoM_PHB05 (accession no. NC_052652) in ICTV. This phage represents a novel viral genus of the subfamily Stephanstirmvirinae, for which we proposed “Sichuanvirus” as the genus name. This phage has a narrow host range lyse specific for KL64 Klebsiella. This phage has no genes referring to antimicrobial resistance, virulence, and lysogen and is stable to a wide range of pH and temperatures. We also obtained three bacterial mutants resistant to the phage and performed genome sequencing for them. We therefore discovered that the interruption of a capsular polysaccharide biosynthesis-related gene wcaJ by insertion sequences mediated the resistance to this phage.ConclusionWe recovered and characterized a phage of “Sichuanvirus”, a novel viral genus of subfamily Stephanstirmvirinae, which is suitable for phage therapy. The discovery of this phage expands the arsenal against CRKP.

Cophylogeny, narrow host breadth and local conditions drive highly specialized bird-haemosporidian associations in West-Central African sky islands.

The parasite island syndrome denotes shifts in parasite life histories on islands, which affect parasite diversity, prevalence and specificity. However, current evidence of parasite island syndromes mainly stems from oceanic islands, while sky islands (i.e. mountains isolated by surrounding low-elevation habitats) have received limited attention. To explore the parasite syndrome in Afrotropical sky islands, we examined haemosporidian blood parasites and their bird hosts in two Afromontane regions in Cameroon. Analysing more than 1300 bird blood samples from the Bamenda Highlands and Mount Cameroon, we found considerably reduced parasite lineage diversity and total prevalence in Mt Cameroon, but not in the Bamenda Highlands. We found highly specific parasite-host interactions at both sites and these associations showed significant phylogenetic congruence, suggesting that closely related parasites infect phylogenetically related hosts. These parasite-host associations tend to be shaped mainly by duplications, switches, losses and failures to diverge rather than through co-speciation events. Overall, the high specificity and parasite-host association differences at local scales largely agree with the limited insights from other sky islands. Yet the drivers of these interactions differ geographically, suggesting that unique dynamics of fragmentation and isolation of montane habitats can lead to similar patterns of host-parasite interactions that are shaped by different underlying drivers.

Genetic variability and population structure analysis of Protostrongylus oryctolagi (Nematoda: Protostrongylidae) in Lepus europaeus from Central and Northern Italy.

Nematodes are abundant and ubiquitous animals which are poorly known at intraspecific level. This work represents the first attempt to fill the gap on basic knowledge of genetic variability and differentiation in Protostrongylus oryctolagi, a nematode parasite of lagomorphs. 68 cox1 sequences were obtained from brown hares collected in five locations in Northern and Central Italy, highlighting the presence of a high amount of genetic variation inside this species. The eleven haplotypes identified (Haplotype diversity equal to 0.702) were split into two lineages: lineage A (comprising six different haplotypes, A1-A6) and lineage B (B1-B5). The mean intra-lineage amount of genetic variation was 0.3%, whereas the inter-lineage percentage of variation was ten-fold higher (3%). These two lineages were non-randomly distributed in the investigated areas. Lineage A showed a preference for Central Italy (Tuscany) even if it was sporadically found also in northern territories (Emilia-Romagna), while B-haplotypes were present exclusively in Emilia-Romagna. The analysis of molecular variance identified two main barriers to gene flow: (i) a strong major one which separate samples of Central Italy (PIA and GR7) from the northern ones (RE1, RE3 and MO1; ΦST = 0.750, P = 0.00); (ii) a secondary faint barrier which separates Pianosa island from Grosseto (ΦST = 0.133, P = 0.00). Any difference was found among northern samples (ΦST = 0.009, P = 0.00). The observed data may be explained by several factors ranging from the parasite's biology (presence of a narrow host spectrum), the final host's behaviour (small home range), the natural dispersion of the host-parasite dyad occurred in past or the recent passive men-mediated migration. Finally, the presence of unconventional shortened amplicons revealed the presence of NUMTs (nuclear copy of mitochondrial genes) in the P. oryctolagi nuclear genome, suggesting caution when using DNA barcode as unique marker for the identification of species belonging to this genus. "In short, if all the matter in the universe except the nematodes were swept away, our world would still be dimly recognizable". Nathan Augustus Cobb, from "Nematodes and Their Relationships", 1915.

Isolation of Bacteriophage Against the Drug Resistant Clinical Pathogens and Host Range Analysis

Antibiotic crisis is a major challenge being encountered by the global population as a result of development of antibiotic-resistant bacteria. Thus, shifting the focus to explore alternative methods to treat bacterial infection. Bacteriophage, the natural predator of the bacteria is one of those alternatives. This study is aimed to isolate bacteriophages from different types of water samples which are capable of infecting drug-resistant clinical isolates and understand its host range capacity. The isolation of bacteriophage was done by Double Layer Agar Assay (DLAA) from 3 different water samples i.e sewage sample from Bagmati and ALKA hospital and from Lele river water. For the isolation of phage, host organisms used were Multi Drug Resistant (MDR) E. coli and K. oxytoca resistant to Amoxyclav, Cefotaxime, Ciproflaxcin, Erythromycin, and Nalidixic Acid. Phage against E. coli II was isolated from both sewage samples while K. oxytoca phage was isolated from Bagamti sewage only. Phage could not be isolated against E. coli I and K. oxytoca phage could not be sub cultured as no plaques were formed on sub culturing and it showed temperate properties. The host range analysis of E. coli II phage showed that the phage has narrow host range as it did not formed plaques against bacteria of other genera. This indicates that sewage is the best source for discovery of phage and the isolated bacteriophage are capable of lytic activity on MDR bacterial isolates. Even though the isolated bacteriophage has potential to be used in phage therapy, still further research should be done for its application. Int. J. Appl. Sci. Biotechnol. Vol 12(4): 201-206.

Engineering Phages to Fight Multidrug-Resistant Bacteria.

Facing the global "superbug" crisis due to the emergence and selection for antibiotic resistance, phages are among the most promising solutions. Fighting multidrug-resistant bacteria requires precise diagnosis of bacterial pathogens and specific cell-killing. Phages have several potential advantages over conventional antibacterial agents such as host specificity, self-amplification, easy production, low toxicity as well as biofilm degradation. However, the narrow host range, uncharacterized properties, as well as potential risks from exponential replication and evolution of natural phages, currently limit their applications. Engineering phages can not only enhance the host bacteria range and improve phage efficacy, but also confer new functions. This review first summarizes major phage engineering techniques including both chemical modification and genetic engineering. Subsequent sections discuss the applications of engineered phages for bacterial pathogen detection and ablation through interdisciplinary approaches of synthetic biology and nanotechnology. We discuss future directions and persistent challenges in the ongoing exploration of phage engineering for pathogen control.

Lytic Spectra of Tailed Bacteriophages: A Systematic Review and Meta-Analysis.

As natural predators of bacteria, tailed bacteriophages can be used in biocontrol applications, including antimicrobial therapy. Also, phage lysis is a detrimental factor in technological processes based on bacterial growth and metabolism. The spectrum of bacteria bacteriophages interact with is known as the host range. Phage science produced a vast amount of host range data. However, there has been no attempt to analyse these data from the viewpoint of modern phage and bacterial taxonomy. Here, we performed a meta-analysis of spotting and plaquing host range data obtained on strains of production host species. The main metric of our study was the host range value calculated as a ratio of lysed strains to the number of tested bacterial strains. We found no boundary between narrow and broad host ranges in tailed phages taken as a whole. Family-level groups of strictly lytic bacteriophages had significantly different median plaquing host range values in the range from 0.18 (Drexlerviridae) to 0.70 (Herelleviridae). In Escherichia coli phages, broad host ranges were associated with decreased efficiency of plating. Bacteriophage morphology, genome size, and the number of tRNA-coding genes in phage genomes did not correlate with host range values. From the perspective of bacterial species, median plaquing host ranges varied from 0.04 in bacteriophages infecting Acinetobacter baumannii to 0.73 in Staphylococcus aureus phages. Taken together, our results imply that taxonomy of bacteriophages and their bacterial hosts can be predictive of intraspecies host ranges.

Isolation, partial characterization, therapeutic, and safety evaluation of carbapenem-resistant Acinetobacter baumannii lytic phage in a mouse model

BackgroundAntimicrobial resistance (AMR) is a major worldwide health concern, characterized by the ability of microorganisms to withstand the effects of medications that once effectively treated infections. Phage therapy has emerged as a promising alternative for management of multidrug-resistant (MDR) bacterial infections. Acinetobacter baumannii (A. baumannii) exemplifies the emergence of bacteria resistant to clinically relevant antimicrobials, leading to severe nosocomial infections and exhibiting extensive and pan drug-resistant (XDR and PDR) traits. In response, this study isolated A. baumannii virulent phage designated as vB_AbaP_PhE54 against carbapenem-resistant A. baumannii (CRAB) pathogen and examined its morphological characteristics using an electron micrograph. Phage stability at different temperatures, pH, chloroform, safety, therapeutic evaluation, and growth kinetics have been analyzed.ResultsThe A. baumannii phage vB_AbaP_PhE54 belongs morphologically to the Podoviridae family with very short, noncontractile tails, the phage demonstrated high thermal tolerance and infectivity across a pH range of 4–11, although it displayed a narrow host range. One-step growth kinetics indicated a burst size of 85 PFU (Plaque Forming Unit) per infected cell and a latent period of 20 min. Additionally, therapeutic efficiency in a mouse model showed total elimination of CRAB pathogen from lungs homogenates of mice and recovery from lung inflammation in all infected mice. On the other hand, safety evaluation of isolated phage revealed no adverse effects on structural or morphological tissue integrity.ConclusionsThese findings suggest that A. baumannii phage vB_AbaP_PhE54 could be a viable safe therapeutic option against A. baumannii infections, warranting further research into its clinical applications.

Isolation and Characterization of Lytic Phages Infecting Clinical Klebsiella pneumoniae from Tunisia.

Background: Klebsiella pneumoniae is an opportunistic pathogen that causes a wide range of infections worldwide. The emergence and spread of multidrug-resistant clones requires the implementation of novel therapeutics, and phages are a promising approach. Results: In this study, two Klebsiella phages, KpTDp1 and KpTDp2, were isolated from wastewater samples in Tunisia. These phages had a narrow host range and specifically targeted the hypervirulent K2 and K28 capsular types of K. pneumoniae. Both phages have double-stranded linear DNA genomes of 49,311 and 49,084 bp, respectively. Comparative genomic and phylogenetic analyses placed phage KpTDp2 in the genus Webervirus, while phage KpTDp1 showed some homology with members of the genus Jedunavirus, although its placement in a new undescribed genus may be reconsidered. The replication efficiency and lytic ability of these phages, combined with their high stability at temperatures up to 70 °C and pH values ranging from 3.5 to 8.2, highlight the potential of these phages as good candidates for the control of hypervirulent multidrug-resistant K. pneumoniae. Methods: Phage isolation, titration and multiplicity of infection were performed. The stability of KpTDp1 and KpTDp2 was tested at different pH and temperatures. Genomic characterization was done by genome sequencing, annotation and phylogenetic analysis. Conclusions: The ability of KpTDp1 and KpTDp2 to lyse one of the most virulent serotypes of K. pneumoniae, as well as the stability of their lytic activities to pH and temperature variations, make these phages promising candidates for antibacterial control.

The evolution of complex metarhizium-insect-plant interactions

Metarhizium species interact with plants, insects, and microbes within a diffuse co-evolutionary framework that benefits soil health, biodiversity, and plant growth. The insect host ranges of these fungi vary greatly. Specialization to a narrow host range usually occurs in the tropics with its stable insect populations, and is characterized by the rapid evolution of existing protein sequences, sexual recombination, and small genomes. Host-generalists are associated with temperate regions and ephemeral insect populations. Their mutualistic plant-colonizing lifestyle increases survival when insects are rare, while facultative entomopathogenicity feeds both the fungi and plants when insects are common. Host-generalists have lost meiosis and associated genome defense mechanisms, enabling gene duplications to diversify functions related to plant colonization and host exploitation. Horizontal gene transfer events via transposons have also contributed to host range changes, while parasexuality combines beneficial mutations within individual clones of host-generalists. There is also a lot of genetic variation in insect populations and an increasing understanding that both pathogen virulence and insect immunity are linked with stress responses. Thus, susceptibility to host-generalists can vary due to non-specific resistance to multiple stressors, multipurpose physical and chemical barriers, and heterogeneity in physiological and behavioral factors, such as sleep.

A blueprint for broadly effective bacteriophage-antibiotic cocktails against bacterial infections

Bacteriophage (phage) therapy is a promising therapeutic modality for multidrug-resistant bacterial infections, but its application is mainly limited to personalized therapy due to the narrow host range of individual phages. While phage cocktails targeting all possible bacterial receptors could theoretically confer broad coverage, the extensive diversity of bacteria and the complexity of phage-phage interactions render this approach challenging. Here, using screening protocols for identifying “complementarity groups” of phages using non-redundant receptors, we generate effective, broad-range phage cocktails that prevent the emergence of bacterial resistance. We also discover characteristic interactions between phage complementarity groups and particular antibiotic classes, facilitating the prediction of phage-antibiotic as well as phage-phage interactions. Using this strategy, we create three phage-antibiotic cocktails, each demonstrating efficacy against ≥96% of 153 Pseudomonas aeruginosa clinical isolates, including biofilm cultures, and demonstrate comparable efficacy in an in vivo wound infection model. We similarly develop effective Staphylococcus aureus phage-antibiotic cocktails and demonstrate their utility of combined cocktails against polymicrobial (mixed P. aeruginosa/S. aureus) cultures, highlighting the broad applicability of this approach. These studies establish a blueprint for the development of effective, broad-spectrum phage-antibiotic cocktails, paving the way for off-the-shelf phage-based therapeutics to combat multidrug-resistant bacterial infections.

Hepatitis B and D virus entry.

Hepatitis B virus (HBV) entry is the initial step of viral infection, leading to the formation of covalently closed circular DNA, which is a molecular reservoir of viral persistence and a key obstacle for HBV cure. The restricted entry of HBV into specific cell types determines the nature of HBV, which has a narrow host range in tissues and species. Hepatitis D virus (HDV) shares viral surface antigens with HBV and thus follows a similar entry mechanism at its early stages. In late 2012, sodium taurocholate cotransporting polypeptide was discovered as an HBV and HDV entry receptor. Since then, the mechanisms of HBV and HDV entry have been extensively analysed. These analyses have expanded our understanding of HBV and HDV host tropism and have provided new strategies for the development of antiviral agents. Notably, the structures of sodium taurocholate cotransporting polypeptide and its interaction with the 2-48 amino acid region of viral preS1 have been recently solved. These findings will stimulate further entry studies. In this Review, we summarize current understanding of HBV and HDV entry and future perspectives.

Characterization and genome analysis of Klebsiella phages with lytic activity against Klebsiella pneumoniae.

Klebsiella pneumoniae is an important gram-negative opportunistic pathogen that causes a variety of infectious diseases. As K. pneumoniae are becoming increasingly resistant to antibiotics, the use of bacteriophages may offer a non-antibiotic-based approach to treat these infections. In the present study, five lytic bacteriophages, 2044307w, k2044hw, k2044ew, k2044302 and 2146hw specific to K. pneumoniae were isolated from hospital sewage and characterized. They belong to group A of the KP32viruses based on transmission electron microscopy (TEM) and genome analysis. These bacteriophages have an extremely narrow host spectrum. The phenotypic characteristics of the phages were determined using lysis assay, pH, and temperature stability tests. This contributes to expanding our understanding of K. pneumoniae phages.

A Klebsiella-phage cocktail to broaden the host range and delay bacteriophage resistance both in vitro and in vivo

Bacteriophages (phages), viruses capable of infecting and lysing bacteria, are a promising alternative for treating infections from hypervirulent, antibiotic-resistant pathogens like Klebsiella pneumoniae, though narrow host range and phage resistance remain challenges. In this study, the hypervirulent K. pneumoniae NTUH-K2044 was used to purify phage ΦK2044, while two ΦK2044-resistant strains were used to purify two further phages: ΦKR1, and ΦKR8 from hospital sewage. A detailed characterization showed that ΦK2044 specifically killed KL1 capsule-type K. pneumoniae, while ΦKR1 and ΦKR8 targeted 13 different capsular serotypes. The phage cocktail (ΦK2044 + ΦKR1 + ΦKR8) effectively killed K. pneumoniae in biofilms, pre-treatment biofilm formation, and delayed phage-resistance. The phage cocktail improved 7-day survival in Galleria mellonella and mouse models and showed therapeutic potential in a catheter biofilm model. In summary, this proof-of-principle phage cocktail has a broad host range, including hypervirulent and highly drug-resistant K. pneumoniae, and serves as a promising starting point for optimizing phage therapy.

Exploring the genomic basis of Mpox virus-host transmission and pathogenesis.

Mpox disease, caused by the monkeypox virus (MPXV), was recently classified as a public health emergency of international concern due to its high lethality and pandemic potential. MPXV is a zoonotic disease that emerged and is primarily spread by small rodents. Historically, it was considered mainly zoonotic and not likely to sustain human-to-human transmission. However, the worldwide outbreak of Clade IIb MPXV from 2020 to 2022 and ongoing Clade I MPXV epidemics in the Democratic Republic of the Congo and surrounding areas are a warning that human-adapted MPXVs will continually arise. Understanding the viral genetic determinants of host range, pathogenesis, and immune evasion is imperative for developing control strategies and predicting the future of Mpox. Here, we delve into the MPXV genome to detail genes involved in host immune evasion strategies for this zoonotic rodent-borne and human-circulating virus. We compare MPXV gene content to related Orthopoxviruses, which have narrow host ranges, to identify potential genes involved in species-specific pathogenesis and host tropism. In addition, we cover the key virulence factor differences that distinguish the MPXV clade lineages. Finally, we dissect how genomic reduction of Orthopoxviruses, through various molecular mechanisms, is contributing to the generation of novel MPXV lineages with increased human adaptation. This review aims to highlight gene content that defines the MPXV species, MPXV clades, and novel MPXV lineages that have culminated in this virus being elevated to a public health emergency of national concern.

Characterization and Potential Application of Phage vB_PmuM_CFP3 for Phage Therapy Against Avian Pasteurella multocida.

The rise of antibiotic-resistant bacterial infections necessitates alternative therapeutic strategies, such as phage therapy. This study investigates the potential of phage vB_PmuM_CFP3 (CFP3) as a therapeutic agent against avian cholera caused by Pasteurella multocida (P. multocida). Phage CFP3 was isolated from the feces and wastewater of a laying hen farm and underwent comprehensive biological characterization, including host range, lytic activity, and environmental stability. Transmission electron microscopy revealed CFP3's typical myovirus morphology, with a head diameter of approximately 60 nm and a tail length of about 120 nm. CFP3 demonstrated high stability across a pH range of 4-10 and temperatures of 30-40 °C, making it suitable for oral administration in poultry. The phage exhibited a latent period of about 90 min and an optimal multiplicity of infection (MOI) of 1. Despite its narrow host range, with a lysis rate of 28.2% against avian-derived type A P. multocida, CFP3's specificity minimizes impact on non-target bacteria. Whole-genome sequencing revealed a 32,696 bp linear double-stranded DNA genome with 46 predicted open reading frames (ORFs) and no tRNA or antibiotic resistance genes, enhancing its safety profile. Phylogenetic analysis indicated a close evolutionary relationship with Haemophilus phages HP1, HP2, and Pasteurella phage F108. While CFP3 shows promise as a precision therapeutic tool, further in vivo studies are required to evaluate its efficacy and safety. Future research should focus on expanding the phage library, optimizing phage mixtures, and exploring synergistic effects with other antimicrobial strategies. This study provides foundational data supporting the development of CFP3 as a viable alternative to antibiotics for controlling avian cholera.

Identification of the critical residues of TMPRSS2 for entry and host range of human coronavirus HKU1.

Human coronavirus (CoV) HKU1 infection typically causes common cold but can lead to pneumonia in children, older people, and immunosuppressed individuals. Recently, human transmembrane serine protease 2 (hTMPRSS2) was identified as the functional receptor for HKU1, but its region and residues critical for HKU1 S binding remain elusive. In this study, we find that HKU1 could utilize human and hamster, but not rat, mouse, or bat TMPRSS2 for virus entry, displaying a narrow host range. Using human-bat TMPRSS2 chimeras, we show that the serine peptidase (SP) domain of TMPRSS2 is essential for entry of HKU1. Further extensive mutagenesis analyses of the C-terminal regions of SP domains of human and bat TMPRSS2s identify residues 417 and 469 critical for entry of HKU1. Replacement of either D417 or Y469 with asparagine in hTMPRSS2 abolishes its abilities to mediate entry of HKU1 S pseudovirions and cell-cell fusion, whereas substitution of N417 with D or N469 with Y in bat TMPRSS2 (bTMPRSS2) renders it supporting HKU1 entry. Our findings contribute to a deeper understanding of coronavirus-receptor interactions and cross-species transmission.IMPORTANCEThe interactions of coronavirus (CoV) S proteins with their cognate receptors determine the host range and cross-species transmission potential. Recently, human transmembrane serine protease 2 (hTMPRSS2) was found to be the receptor for HKU1. Here, we show that the TMPRSS2 of hamster, but not rat, mouse, or bat, can serve as a functional entry receptor for HKU1. Moreover, swapping the residues at the positions of 417 and 469 of bTMPRSS2 with the corresponding residues of hTMPRSS2 confers it supporting entry of HKU1 S pseudovirions, indicating the critical role of these residues in HKU1 entry. Our study identified the critical residues in hTMPRSS2 responsible for receptor interaction and host range of HKU1.

E2F3-dependent activation of FAM111B restricts mouse cytomegalovirus replication in primate cells.

Viruses must counteract host cell defenses to facilitate viral replication. Viruses with a narrow host range, such as the cytomegaloviruses, are unable to counteract cellular defenses in cells of a foreign species. However, little is known about the cellular host range factors restricting cytomegalovirus replication. Here, we show that mouse cytomegalovirus (MCMV) induces the expression of the FAM111 proteases and that FAM111B, but not FAM111A that has previously been shown to restrict the replication of polyomavirus and orthopoxvirus host range mutants, acts as a cellular factor suppressing MCMV replication in human and rhesus monkey cells. The identification of FAM111B as a host range factor should provide new insight into the physiological functions of this poorly characterized protein.

Seasonal succession, host associations, and biochemical roles of aquatic viruses in a eutrophic lake plagued by cyanobacterial blooms

Viruses are implicated to play key roles as biogeochemical mediators and ecological drivers in freshwater ecosystems. However, the dynamics of viruses and host associations throughout the seasons and during blooming periods in eutrophic freshwater ecosystems remain poorly understood. From the water microbiomes of planktonic biomass from Lake Taihu, a large eutrophic freshwater lake in China that experiences annual Microcystis-dominated harmful algal blooms (HABs), we recovered 41,997 unique viral clusters spanning a wide taxonomic range, including 15,139 Caudovirales clusters targeting bacteria and 1,044 NCLDV clusters targeting eukaryotes. The viral community exhibited clear seasonal succession, driven primarily by microbial communities (particularly Cyanobacteria and Planctomycetes) and environmental factors (mainly nutrients and temperature). Host prediction revealed that viral infection had a more distinct impact on bacteria-driven nitrogen pathways than on phosphate cycling. HAB-induced variations in microbial composition and environmental conditions affected viral strategies including viral lifestyles, host range, and virus-encoded auxiliary metabolic genes (vAMGs) distributions. Viruses infecting Proteobacteria and Actinobacteria showed an enhanced lysogenic lifestyle and a narrower host range during HAB peak in summer, while viruses infecting Bacteroidota adopted an opposite strategy. Notably, vAMGs were most abundant before the HAB outbreak in spring, compensating for bacterial metabolic processes of their hosts such as carbohydrates metabolism, photosynthesis, and phosphate regulation. The findings highlight the intricate relationships between viruses, host microbes, and the bloom-associated environment, underscoring the important biochemical roles viruses play in eutrophic freshwater ecosystems.

Reconstructing host plant repertoire and timing of evolution of phyline plant bugs (Hemiptera, Miridae)

AbstractThe diversity of phytophagous insects is often attributed to the success of land plants in the framework of ecological speciation. Several hypotheses have been proposed to explain host plant driven insect diversification in a phylogenetic context and have mostly been explored using Lepidoptera. We posit that Miridae are a great system to examine these hypotheses because they are one of the largest primarily phytophagous insect families and include many species with narrow host repertoire. Focusing on the species‐rich Phylinae (>2700 spp.), we generate the most taxon‐rich phylogeny published to date and for the first time estimate divergence times and trace the evolution of host plant associations across the group. Focusing on two clades of oak‐associated phylines, we further examine if diversification in these insects and their hosts coincided or if the insects tracked their hosts. We find that Phylinae diverged from their orthotyline sister group before the end of the Cretaceous, tribal‐level taxa diversified throughout the Paleogene, and diversification within genera mostly occurred in the Neogene. Host plant repertoire reconstructions at the family level show transitions from stenophagy to polyphagy are more common than the reverse. We reconstructed the ancestral phyline host as ambiguous, followed by Asterales throughout most of the deep splits. Species‐level divergences in the two oak‐associated clades coincide with those in oaks, a pattern is that is consistent with the hypothesis that these plant bugs may have cospeciated with their hosts. Our study shows that Phylinae are a suitable system to further test hypotheses on ecological speciation of plants and insects but will require more robust phylogenetic hypotheses of the group.

Host population structure and species resolution reveal prophage transmission dynamics.

Much knowledge about bacteriophages has been obtained via genomics and metagenomics over the last decades. However, most studies dealing with prophage diversity have rarely conducted phage species delimitation (aspect 1) and have hardly integrated the population structure of the host (aspect 2). Yet, these two aspects are essential in assessing phage diversity. Here, we implemented an operational definition of phage species (clustering at 95% identity, 90% coverage) and integrated the host's population structure to understand prophage diversity better. Gathering the most extensive data set of Acinetobacter baumannii phages, we show that most prophage species have four or fewer prophages per species, and just five prophage species have more than 100 prophages. Most prophage species have a narrow host range and are geographically restricted; yet, very few have a broad host range being well spread in distant lineages of A. baumannii. These few broad host range prophage species are cosmopolitan and the most abundant species. Prophages in the same bacterial genome are very divergent, and prophages can easily be gained and lost within the bacterial lineages. Finally, even with this extensive data set, the prophage diversity has not been fully grasped. This study shows how integrating the host population structure and clustering at the species level allows us to better appreciate phage diversity and its transmission dynamics.