Primary Host Research Articles

Effects of reductive soil disinfestation on potential pathogens and antibiotic resistance genes in soil

Reductive soil disinfestation (RSD) is commonly employed for soil remediation in greenhouse cultivation. However, its influence on antibiotic resistance genes (ARGs) in soil remains uncertain. This study investigated the dynamic changes in soil communities, potential bacterial pathogens, and ARG profiles under various organic material treatments during RSD, including distillers’ grains, potato peel, peanut vine, and peanut vine combined with charcoal. Results revealed that applying diverse organic materials in RSD significantly altered bacterial community composition and diminished the relative abundance of potential bacterial pathogens (P < 0.05). The relative abundance of high-risk ARGs decreased by 10.7%-30.6% after RSD treatments, the main decreased ARG subtypes were AAC(3)_Via, dfrA1, ErmB, lnuB, aadA. Actinobacteria was the primary host of ARGs and was suppressed by RSD. Soil physicochemical properties, such as total nitrogen, soil pH, total carbon, were crucial factors affecting ARG profiles. Our findings demonstrated that RSD treatment inhibited pathogenic bacteria and could be an option for reducing high-risk ARG proliferation in soil.

Prevalence and dissemination of mcr-9.1-producing non-typhoidal Salmonella strains from diarrhea patients throughout China during 2010–2020

The emergence of mobilized colistin resistance (mcr) genes has raised significant concerns as they pose a public health issue. The prevalence of mcr genes, particularly the newly discovered mcr-9 gene, in non-typhoidal Salmonella (NTS) isolates remains unclear. We characterized mcr-9.1-producing NTS isolates from China. Among 7,106 NTS isolates from diarrhea cases in 32 provinces during 2010–2020, 11 mcr-9.1-producing isolates were identified and were all not resistant to colistin. Five isolates belonged to Salmonella Thompson and sequence type (ST) 26, two belonged to Salmonella Typhimurium and ST34, two belonged to Salmonella Typhimurium and ST36, and two belonged to Salmonella 1,4,[5],12:i:- and ST34. Plasmids harboring mcr-9.1 tended to possess the IncHI2 backbone and were ~ 300 kb long. All mcr-9.1 genes shared the same flanking sequence, rcnR-rcnA-pcoS-IS903-mcr-9.1-wbuC. According to the NCBI data, we found that NTS serves as the primary host of mcr-9.1, although the prevalence of specific serotypes differed between domestic and international settings. Notably, most data came from developed countries, such as the USA. mcr-9.1 tended to be transferred as a gene cassette or to be mobilized by a conjugational plasmid in multiple bacteria across humans, animals, and the environment. Furthermore, mcr-9.1 frequently co-existed and was co-transferred with various genes encoding resistance to first-line drugs, reducing the effectiveness of available therapeutic options. In summary, although mcr-9 does not mediate colistin resistance, it can silently spread with some genes encoding resistance to first-line drugs, and therefore warrants research attention.

Burdening Perspectives and Treatment Modalities of Monkeypox: A Central Dogma.

The monkeypox virus (MPXV), belonging to the genus Orthopoxvirus, is responsible for causing the zoonotic illness known as Monkeypox. The virus was initially identified during an outbreak at a Danish Zoo in 1958 and has since been found to infect various mammal species worldwide. While African squirrels and other rodents are believed to be the primary hosts, determining the natural host has proven challenging. While MPXV can be studied using different animal models in laboratory settings, understanding its natural transmission routes remains complex and species-dependent. Recent developments have elevated the global health concern surrounding Monkeypox, leading to its designation as a Global Health Emergency of International Concern on 23 July 2022. Enhancing surveillance and case detection is crucial in navigating the unpredictable epidemiology of this re-emerging disease. Human infections with the monkeypox virus are becoming less frequent due to population growth and economic improvements. Monkeypox, similar to smallpox, can potentially be controlled and eradicated in the future through vaccines, appropriate treatment, and personal protective equipment.

A Microcosm Experiment Reveals the Temperature-Sensitive Release of Mucochytrium quahogii (=QPX) from Hard Clams and Pallial Fluid as a Stable QPX Reservoir.

Mucochytrium quahogii, also known as QPX or Quahog Parasite Unknown, is the causative agent of QPX disease in the hard clam (Mercenaria mercenaria). Host-pathogen-environment interactions between M. quahogii, the hard clam, and temperature were explored in a microcosm experiment. Hard clams were housed in individual tanks with sterile seawater under two temperature regimes: low (13 °C) temperature, which is thought to be optimal for QPX disease development, and high (20 °C) temperature, which has been shown to promote "healing" of QPX-infected clams. Hard clam tissue, pallial fluid, seawater, and shell biofilms were collected and assayed for M. quahogii. The release of M. quahogii from naturally infected live hard clams into seawater was detected only in the low temperature treatment, suggesting that temperature influences the release of potentially infectious cells. M. quahogii was commonly found in hard clam pallial fluid, even after 9 weeks in the lab, suggesting pallial fluid is a stable reservoir of M. quahogii within its primary host and that M. quahogii is not a transient component of the hard clam microbiota. Overall, results support a host-specific relationship and that M. quahogii is a commensal member of the hard clam microbiota, supporting its classification as an opportunistic pathogen.

Updated Insights into the Phylogenetics, Phylodynamics, and Genetic Diversity of Nipah Virus (NiV).

Nipah virus (NiV), a biosafety level 4 agent, was first identified in human clinical cases during an outbreak in 1998 in Malaysia and Singapore. While flying foxes are the primary host and viral vector, the infection is associated with a severe clinical presentation in humans, resulting in a high mortality rate. Therefore, NiV is considered a virus with an elevated epidemic potential which is further underscored by its recent emergence (September 2023) as an outbreak in India. Given the situation, it is paramount to understand the molecular dynamics of the virus to shed more light on its evolution and prevent potential future outbreaks. In this study, we conducted Bayesian phylogenetic analysis on all available NiV complete genomes, including partial N-gene NiV sequences (≥1000 bp) in public databases since the first human case, registered in 1998. We observed the distribution of genomes into three main clades corresponding to the genotypes Malaysia, Bangladesh and India, with the Malaysian clade being the oldest in evolutionary terms. The Bayesian skyline plot showed a recent increase in the viral population size since 2019. Protein analysis showed the presence of specific protein families (Hendra_C) in bats that might keep the infection in an asymptomatic state in bats, which also serve as viral vectors. Our results further indicate a shortage of complete NiV genomes, which would be instrumental in gaining a better understanding of NiV's molecular evolution and preventing future outbreaks. Our investigation also underscores the critical need to strengthen genomic surveillance based on complete NiV genomes that will aid thorough genetic characterization of the circulating NiV strains and the phylogenetic relationships between the henipaviruses. This approach will better prepare us to tackle the challenges posed by the NiV virus and other emerging viruses.

NO3− as an electron acceptor elevates antibiotic resistance gene and human bacterial pathogen risks in managed aquifer recharge (MAR): A comparison with O2

Managed aquifer recharge (MAR) stands out as a promising strategy for ensuring water resource sustainability. This study delves into the comparative impact of nitrate (NO3−) and oxygen (O2) as electron acceptors in MAR on water quality and safety. Notably, NO3−, acting as an electron acceptor, has the potential to enrich denitrifying bacteria, serving as hosts for antibiotic resistance genes (ARGs) and enriching human bacterial pathogens (HBPs) compared to O2. However, a direct comparison between NO3− and O2 remains unexplored. This study assessed risks in MAR effluent induced by NO3− and O2, alongside the presence of the typical refractory antibiotic sulfamethoxazole. Key findings reveal that NO3− as an electron acceptor resulted in a 2 times reduction in dissolved organic carbon content compared to O2, primarily due to a decrease in soluble microbial product production. Furthermore, NO3− significantly enriched denitrifying bacteria, the primary hosts of major ARGs, by 747%, resulting in a 66% increase in the overall abundance of ARGs in the effluent of NO3− MAR compared to O2. This escalation was predominantly attributed to horizontal gene transfer mechanisms, as evidenced by a notable 78% increase in the relative abundance of mobile ARGs, alongside a minor 27% rise in chromosomal ARGs. Additionally, the numerous denitrifying bacteria enriched under NO3− influence also belong to the HBP category, resulting in a significant 114% increase in the abundance of all HBPs. The co-occurrence of ARGs and HBPs was also observed to intensify under NO3− influence. Thus, NO3− as an electron acceptor in MAR elevates ARG and HBP risks compared to O2, potentially compromising groundwater quality and safety.

Molecular epidemiology and phylogeny of the emerging zoonotic virus Rocahepevirus: A global genetic analysis

Human infections with Rocahepevirus ratti genotype C1 (HEV-C1) in Hong Kong of China, Canada, Spain, and France have drawn worldwide concern towards Rocahepevirus. This study conducted a global genetic analysis of Rocahepevirus, aiming to furnish comprehensive molecular insights and promote further research. We retrieved 817 Rocahepevirus sequences from the GenBank database through October 31, 2023, categorizing them according to research, sample collection area and date, genotype, host, and sequence length. Subsequently, we conducted descriptive epidemiological, phylogenetic evolutionary, and protein polymorphism (in length and identity) analyses on these sequences. Rocahepevirus genomes were identified across twenty-eight countries, predominantly in Asia (71.73%, 586/817) and Europe (26.44%, 216/817). The HEV-C1 dominates Rocahepevirus (77.2%, 631/817), while newly discovered Rocahepevirus genotypes (C3/C4/C5 and other unclassified genotypes) were primarily identified in Europe (25/120) and China (91/120). Muridae animals (72.5%, 592/817) serve as the primary hosts for Rocahepevirus, with other hosts encompassing species from the families Soricidae, Hominidae, Mustelidae, and Cricetidae. Additionally, Rocahepevirus genomes (C1 genotype) were identified in sewage samples recently. The phylogenetic evolution of Rocahepevirus exhibits considerable variation. Specifically, HEV-C1 can be classified into at least six genetic groups (G1 to G6), with human HEV-C1 distributed across multiple evolutionary clades. The overall ORF1 and ORF2 amino acid sequence lengths were significantly different (P < 0.001) across Rocahepevirus genotypes. HEV-C1/C2/C3 and HEV-C4/C5 displayed substantial differences in amino acid sequence identity (58.4%–59.6%). The identification of Rocahepevirus genomes has expanded across numerous countries, particularly in European and Asian countries, coinciding with an expanding host range and emergence of new genotypes. The evolutionary path of Rocahepevirus is intricate, where the HEV-C1 dominates globally and internally forms multiple evolutionary groups (G1 to G6), exhibiting diverse genetic variation within human HEV-C1. Significant differences exist in the protein polymorphism (in length and identity) across Rocahepevirus genotypes. Given Rocahepevirus's shift from an animal virus to a zoonotic pathogen, worldwide cooperation in monitoring Rocahepevirus genomes is vital.

Rohingya refugees and its impact on informal economy: Cox's Bazar, Bangladesh

Purpose: The purpose of this study is to determine whether refugees in Cox's Bazar area of Bangladesh can successfully integrate into the informal economic sector. This evaluation was carried out by comparing the actions of refugees in the labor market with those of the local population.&#x0D; Research methodology: This study used survey data from two population groupings. The sample size included 300 Rohingya refugees and 300 locals from Coxs' Bazar, where they are now living. ANOVA was employed to compare the category mean differences owing to the limited sample size. The calculations would include wealth, occupation, language, religion, and race as labor market barriers.&#x0D; Results: The data indicated significant disparities in both types of occupations and monthly incomes between local workers and refugee workers. Furthermore, the findings also suggest that Rohingya refugees encounter greater challenges when entering the job market than local workers.&#x0D; Limitations: Refugees in Bangladesh are unable to work. They must stay at local and international NGO (Ministry of Foreign Affairs 2014). No Bangladeshi labor legislation has protected them. Bangladeshi authorities can also imprison refugees on illicit travel. A good Rohingya refugee policy in Bangladesh is crucial to human rights.&#x0D; Contribution: The Rohingya Muslim population in Myanmar has been subjected to genocide, resulting in their expulsion and subsequent migration to neighboring countries. Bangladesh is the primary host nation for the Rohingya group from Myanmar, which has sought safety there as refugees for many years. Due to their lack of legal employment opportunities in Bangladesh, these refugees engage in informal economic operations and participate in various criminal activities inside the place where they reside.

Development of a novel tyrosine-based selection system for generation of recombinant Chinese hamster ovary cells

Efficiently expanding Chinese hamster ovary (CHO) cells, which serve as the primary host cells for recombinant protein production, have gained increasing industrial significance. A significant hurdle in stable cell line development is the low efficiency of the target gene integrated into the host genome, implying the necessity for an effective screening and selection procedure to separate these stable cells. In this study, the genes of phenylalanine hydroxylase (PAH) and pterin 4 alpha carbinolamine dehydratase 1 (PCBD1), which are key enzymes in the tyrosine synthesis pathway, were utilized as selection markers and transduced into host cells together with the target genes. This research investigated the enrichment effect of this system and advanced further in understanding its benefits for cell line development and rCHO cell culture. A novel tyrosine-based selection system that only used PCBD1 as a selection marker was designed to promote the enrichment effect. Post 9 days of starvation, positive transductants in the cell pool approached 100%. Applied the novel tyrosine-based selection system, rCHO cells expressing E2 protein were generated and named CHO TS cells. It could continue to grow, and the yield of E2 achieved 95.95mg/L in a tyrosine-free and chemically-defined (CD) medium. Herein, we introduced an alternative to antibiotic-based selections for the establishment of CHO cell lines and provided useful insights for the design and development of CD medium.

Impact of spatial and temporal resource distribution on rabies dynamics in the Arctic

In the Arctic, rabies is endemic in the Arctic fox (Vulpes lagopus), posing a significant and ongoing health risk for people and domestic animals. The mechanisms by which rabies is maintained within the low-density fox populations in the Arctic remain unclear. In this study, we developed a spatially explicit individual-based stochastic epidemiological model and performed an uncertainty analysis to better understand Arctic fox rabies dynamics. Rabies persisted in 25.68% of model simulations, with several variables having significant impact on rabies persistence: probability of rabies transmission, spatial and temporal distribution food resources, mean litter size and variability of rabies incubation periods. Where rabies is endemic, we identified 5 key parameters for rabies dynamics: spatiotemporal resource distribution, probability of birth for adult females, mean and standard deviation of litter size, and incubation period of rabies. Our study demonstrates that Arctic rabies can persist in its primary host under conditions consistent with existing empirical data in the literature and showed the important role played by the spatial and temporal distribution of resources. Finally, our results suggest that the ecological impacts of rapid climate warming could decrease the overall persistence of rabies in the Arctic and the associated health risk in Arctic communities.

From molecules to ecosystems: Insights into a network of interactions for a Chagas disease outbreak using Triatoma brasiliensis as natural samplers

Exploring the dynamics of disease transmission involves an understanding of complex interactions within the eco-epidemiologic framework. In the context of Chagas disease (CD), elements are mainly represented by the interactions among the pathogen, insect vector, host, humans and the environment. We performed quantitative and qualitative analyses on a dataset derived from 98 Triatoma brasiliensis infected by trypanosomatids, which were linked to a CD outbreak in the semi-arid region of northeastern Brazil. We extracted invertebrate-derived DNA (iDNA) from these insects, comprising 18 populations around the outbreak area, each indicative of various strata of anthropogenic influence. Food source (FS) diversity, representing potential parasite reservoirs, was determined through mitochondrial gene (cyt b) sequencing of vertebrates, and parasite genotyping was accessed using fluorescent amplified fragment barcodes (FFLB) of trypanosomatids. We also assessed the residents' awareness of breeding sites for CD vectors in the inspected houses. The quantification of Trypanosoma cruzi was estimated via real-time PCR and is denominated here as the average parasite load (PL) per insect (T. cruzi/intestinal unit). We aimed to address vector-parasite-host-environment interactions that were discussed based on their significance among the components. Notably, among the significant interactions, we observed that the PL in the insects was significantly influenced by FS. Infected insects that fed on the classic reservoir, Didelphis albiventris, and Galea spixii exhibited higher PLs, compared to those that fed on Kerodon rupestris (p < 0.04)—a primary host. While D. albiventris is already recognized as a synanthropic species, we propose that G. spixii may also be undergoing a synanthropic process. Conversely, domestic cats are frequently identified as FS in infected insects from the sylvatic environment, suggesting a possible change in their behavior towards a wild state. Therefore, we propose that neglected anthropogenic actions have facilitated the reciprocal (sylvatic-peridomestic) circulation of T. cruzi—especially noted for TcI because it was predominant in insects found in peridomestic environments. Residents are often unaware of the existence of insect breeding grounds near their homes, particularly when it involves the storage of materials without planning for use, such as piles of tiles, bricks and wood. Although indirect inferences about the interaction among vector-parasite-host-environment are still incipient, we highlight the potential use of vectors as natural samplers of biological and ecological components in transmitting the disease.

LdCyPA attenuates MAPK pathway to assist Leishmania donovani immune escape in host cells

BackgroundVisceral leishmaniasis is a neglected tropical disease affecting millions of people worldwide. Macrophages serve as the primary host cells for L. donovani, the immune response capability of these host cells is crucial for parasites’ intracellular survival. L. donovani peptidyl-prolyl cis/trans isomerase Cyclophilin A (LdCypA) is a key protein for L. donovani intracellular proliferation, while the molecular mechanism conducive to intracellular survival of parasites remains elusive. MethodsIn this study, we generated a macrophage cell line overexpressing LdCyPA to investigate its role in controlling host immunity and promoting intracellular immune escape of L. donovani. ResultsIt was discovered that the overexpression of the LdCyPA cell line regulated the host immune response following infection by downregulating the proportion of M1-type macrophages, promoting the secretion of the anti-inflammatory factor IL-4, and inhibiting the secretion of pro-inflammatory factors like IL-12, IFN-γ, TNF-α, and INOS. Transcriptome sequencing and mechanistic validation, meanwhile, demonstrated that cells overexpressing LdCyPA controlled the immune responses that followed infection by blocking the phosphorylation of P38 and JNK1/2 proteins in the MAPK signaling pathway and simultaneously increasing the phosphorylation of ERK proteins, which helped the L. donovani escape immune recognition. ConclusionOur findings thus pave the way for the development of host-directed antiparasitic drugs by illuminating the pro-Leishmania survival mechanism of L. donovani cyclophilin A and exposing a novel immune escape strategy for L. donovani that targets host cellular immune regulation.

Potential environmental risks of field bio/non-degradable microplastic from mulching residues in farmland: Evidence from metagenomic analysis of plastisphere

The plastisphere may act as reservoir of antibiotic resistome, accelerating global antimicrobial resistance dissemination. However, the environmental risks in the plastisphere of field microplastics (MPs) in farmland remain largely unknown. Here, antibiotic resistance genes (ARGs) and virulence factors (VFs) on polyethylene microplastics (PE-MPs) and polybutylene adipate terephthalate and polylactic acid microplastics (PBAT/PLA-MPs) from residues were investigated using metagenomic analysis. The results suggested that the profiles of ARG and VF in the plastisphere of PBAT/PLA-MPs had greater number of detected genes with statistically higher values of diversity and abundance than soil and PE-MP. Procrustes analysis indicated a good fitting correlation between ARG/VF profiles and bacterial community composition. Actinobacteria was the major host for tetracycline and glycopeptide resistance genes in the soil and PE-MP plastisphere, whereas the primary host for multidrug resistance genes changed to Proteobacteria in PBAT/PLA-MP plastisphere. Besides, three human pathogens, Sphingomonas paucimobilis, Lactobacillus plantarum and Pseudomonas aeruginosa were identified in the plastisphere. The PE-MP plastisphere exhibited a higher transfer potential of ARGs than PBAT/PLA-MP plastisphere. This work enhances our knowledge of potential environmental risks posed by microplastic in farmland and provides valuable insights for risk assessment and management of agricultural mulching applications.

Infestation patterns of a major wood boring pest, Psiloptera fastuosa (Buprestidae: Coleoptera) in Tasar, Terminalia arjuna (Myrtales: Combretaceae) plantation

Tasar silk is a significant cash crop in the tropics. In Tasar culture, arjuna (Terminalia arjuna) (Combretaceae) are often planted on farms because the Tasar worm largely consumes their leaves, Antheraea paphia (Lepidoptera: Saturniidae). The wood-boring Psiloptera fastuosa (Coleoptera: Buprestidae) causes severe threats to Tasar plantations during their mating season (September equinox to December solstice). After hatching, the coleopteran grubs puncture and penetrate the arjuna stem, mature into pupae inside, pass into adults, and exit through emergence holes, causing severe damage to younger plants. Though such infestations are highly fatal, no precautions have been proposed to protect the arjuna plantations. Therefore, sustainable Tasar plantation management is highly challenging because of limited bio-ecological information and the infestation pattern of the pest. This study investigates the origins and implications of P. fastuosa infestations in T. arjuna plantations. Our study focuses on seasonal changes in P. fastuosa assaults on its primary host and infestation frequency in an Indian agroecosystem. Second, we examined whether host plant age and size affected the P. fastuosa invasion. The occurrence and distribution of emergence holes for adult beetles on the arjuna bark and the intensity of wood galleries on the stem aid in assessing the beetle infestation. It was observed that 4 to 8-year-old arjuna plants wilted more frequently than younger (< 4 years) and older (> 8 years) plants. Since most infestations occur on the main trunks up to the middle of the plant's height during autumn, insecticidal treatments can be performed around the middle of the main trunks of 4–8-year-old arjuna plants during fall. The findings of this study will benefit the management and propagation of arjuna plants for tropical Tasar silk production.

Influence of colloidal silicon dioxide‑magnesium stearate interaction on flow and compaction behavior of an MCC-Lactose binary mixture

The amount of glidant and lubricant in a tablet formulation is driven by their primary functionalities of enhancing flowability and reducing particle-wall friction. However, their interaction can lead to undesirable outcomes. This study has examined the influence of this interaction on powder flow and compaction, using colloidal silicon dioxide as a glidant and magnesium stearate as a lubricant in a 50:50 w/w MCC:Lactose binary blend. The findings suggest that nano-sized CSD particles coat other particles in a blend. However, increasing CSD decreases exposed MgSt and reduces its lubricity, resulting in intriguing powder flow and compaction phenomena. CSD impacts powder flow more than MgSt. Increasing the CSD reduces exposed MgSt and increases particle-wall adhesion. Higher CSD concentrations decreased powder bed bulk density and permeability due to the formation of porous structures on primary host particles. A complex interplay between CSD and MgSt during tablet compaction affected tensile strength. CSD promoted tablet picking due to reduced exposed MgSt and increased tablet-punch adhesion. This is the first study correlating tablet picking with take-off force, and it was found that >5 N take-off force risked picking. Ejection force increased with CSD and decreased with MgSt, reaffirming CSD's impact on particle-wall adhesion.

A chromosome-level genome assembly of the Rhus gall aphid Schlechtendalia chinensis provides insight into the endogenization of Parvovirus-like DNA sequences

The Rhus gall aphid, Schlechtendalia chinensis, feeds on its primary host plant Rhus chinensis to induce galls, which have economic importance in medicines and the food industry. Rhus gall aphids have a unique life cycle and are economically beneficial but there is huge gap in genomic information about this group of aphids. Schlechtendalia chinensis induces rich-tannin galls on its host plant and is emerging as a model organism for both commercial applications and applied research in the context of gall production by insects. Here, we generated a high-quality chromosome-level assembly for the S. chinensis genome, enabling the comparison between S. chinensis and non-galling aphids. The final genome assembly is 344.59 Mb with 91.71% of the assembled sequences anchored into 13 chromosomes. We predicted 15,013 genes, of which 14,582 (97.13%) coding genes were annotated, and 99% of the predicted genes were anchored to the 13 chromosomes. This assembly reveals the endogenization of parvovirus-related DNA sequences (PRDs) in the S. chinensis genome, which could play a role in environmental adaptations. We demonstrated the characterization and classification of cytochrome P450s in the genome assembly, which are functionally crucial for sap-feeding insects and have roles in detoxification and insecticide resistance. This genome assembly also revealed the whole genome duplication events in S. chinensis, which can be considered in comparative evolutionary analysis. Our work represents a reference genome for gall-forming aphids that could be used for comparative genomic studies between galling and non-galling aphids and provides the first insight into the endogenization of PRDs in the genome of galling aphids. It also provides novel genetic information for future research on gall-formation and insect-plant interactions.

Spatiotemporal and Species‐Crossing Transmission Dynamics of Subclade 2.3.4.4b H5Nx HPAIVs

Subclade 2.3.4.4b H5Nx highly pathogenic avian influenza (HPAI) viruses, emerged in 2013 with multiple subtypes of H5N8, H5N1, and H5N6, had unprecedently caused a global epizootic by H5N1 since 2021, which had devasted multiple species of wild birds, poultry, and wild mammals (terrestrial and marine) with a high mortality, causing severe ecological damage. The infected wild mammals may become new “mixers” for influenza viruses, posing the potential transmission to human. Frequent outbreaks of subclade 2.3.4.4b H5Nx viruses among wild birds and poultry had exposed major gaps in our knowledge on their evolution, spatiotemporal diffusion, and species‐crossing transmission. Here, we integrated the phylogenetic and epidemiological data of subclade 2.3.4.4b H5Nx viruses in public database and used Bayesian phylodynamic analysis to reveal the pattern of the global large‐scale transmission. Phylogenic analysis demonstrated that the HA gene of these viruses diverged into two dominant clusters around 2015 and 2016. The Bayesian phylodynamic analysis illustrated that the viruses presented spatiotemporally complex transmission network with geographical and host relative expansion and recombination with different subtypes of NA segment. Spatially, the Russian Federation (Siberia) was identified as the primary hub for virus transmission, which was further facilitated by the establishment of strong epidemiological linkages between West Europe and broader regions, such as North America. As for hosts, wild Anseriformes were the primary species for the virus spillover, contributing to the spatial expansion and rapid diffusion globally of subclade 2.3.4.4b viruses. We investigated the phylogeny of subclade 2.3.4.4b H5Nx viruses and the spatiotemporal pattern of transmission with initial location and the primary host, which could provide comprehensive insights for subclade 2.3.4.4b H5Nx viruses. Due to the wild birds involved the widespread of subclade 2.3.4.4b H5Nx viruses, the epizootics in poultry are inevitable, so we highly recommend to apply the policy of culling plus with vaccination to protect the poultry industry and potentially protect the public health.

Studies of Serrasentis sagittifer (Linton, 1889) Van Cleave, 1923 from cobia Rachycentron canadum (Linn.) in the Arabian Gulf; with Molecular Analysis and Optical Microscopy

The finding of a population of Serrasentis sagittifer (Linton, 1889) Van Cleave, 1923 in Cobia Rachycentron canadum (Linn.) in the Arabian Gulf off Iranian waters prompted the study of its molecular parameters known previously only from a population of adults from the same host species, and of immatures from northern Australian waters and from other hosts in the Red Sea. We have previously described S. sagittifer from Cobia and from paratenic hosts in the same Gulf waters off Kuwait and Qatar and detailed its worldwide morphological variability and geographical distribution. We have processed and sampled a few specimens from the Arabian Gulf off Iran for morphological verification of their diagnosis and offered a few light microscopical images that accentuate their identity in Iranian waters for the first time. We document additional diagnostic features of S. sagittifer using optical microscopy. We determine that the distribution of S. sagittifer runs parallel that of its primary adult host, R. canadum, worldwide. We have used the 18S and 28S gene regions of ribosomal RNA, including the mitochondrial cox1 gene, for molecular analysis of an Arabian Gulf population for the first time. Maximum likelihood and Bayesian inference methods were used for phylogenetic reconstructions, the sequences of S. sagittifer from Iran were grouped within the genus Serrasentis and placed as a sister group to S. nadakali George and Nadakal, 1978. Due to the lack of molecular data for other species of this group of Rhadinorhynchidae, the present phylogenetic inferences are required to continue exploring the genetic diversity of this group of species to infer their phylogeny. The cox 1 haplotype network inferred with 14 sequences of S. sagittifer revealed 13 haplotypes separated from each other by a few substitutions. Our ocular microscopy accentuated morphological details of S. Sagittifer and the haplotype network genealogy produced in this study based on mt cox1 sequences from S. sagitiffer show a phylogeographic structure for the first time; therefore, the haplotypes grouped into their own geographic clusters.

Diversity of genes encoding immune-related GTPase B2 protein, an inherited element responsible for resistance against virulent Toxoplasma gondiistrains, among wild Mus musculus in local area of Japan.

The major genetic group of Toxoplasma gondii, known as type I, generally displays high lethality in laboratory Mus musculus (mouse) strains, with few exceptions. However, because rodents are the primary reservoir hosts for T. gondii, if this characteristic manifests in the wild, type I strains would be extinct. Therefore, we hypothesized that populations of wild rodents capable of harboring type I T. gondii asymptomatically exist globally and are not limited to a few localized areas, as previously thought. The strength of mouse resistance to T. gondii is known to depend on the affinity of the mouse-expressed immunity-related GTPases B2 (IRGB2) protein for the T. gondii-expressed rphoptry protein 5B (ROP5B) protein. Therefore, the Irgb2 gene sequences of 12 individuals mice captured at two animal farms in Gifu Prefecture, and on an island in Okinawa Prefecture, Japan were determined, and subjected to a molecular phylogenetic analysis together with those of various mouse strains worldwide. The Irgb2 gene of M. musculus individuals captured on one farm and one island showed diverse sequences. The sequences from two individual mice captured in an animal farm formed a single clade with a wild mouse derived CAST/EiJ strain, known for its exceptional resistance to type I T. gondii lethality. These results suggest that M. musuculus individuals resistant to the Type I T. gondii strain may be present in Japan, in addition to the previously known populations in South Asia, Thailand and India.

Parasitism on domestic cats by Amblyomma auricularium and serological evidence of exposure to Rickettsia amblyommatis.

The domestic cat is not considered a primary host for any specific tick species; however, it can be affected by some Ixodidae species, such as Rhipicephalus sanguineus sensu lato and Amblyomma spp. The study reports parasitism by Amblyomma auricularium and the detection of anti-Rickettsia spp. antibodies in domestic cats from a rural property in the Afrânio municipality, Pernambuco, Brazil. Amblyomma auricularium (24 nymphs, six females, and four males) and Amblyomma sp. (42 larvae) parasitized three cats, and 73 free-living ticks were captured in armadillo burrows: A. auricularium (36 nymphs, six females, five males) and Amblyomma sp. (26 larvae). Blood samples from cats were collected and the obtained plasma were subjected to indirect immunofluorescence assay (IFA) to detect antibodies against Rickettsia antigens. Thus, anti-Rickettsia spp. antibodies were determined (titers ranging from 128 to 512) and showed a predominant antibody response to Rickettsia amblyommatis or a very closely related genotype. This study reports the first infestation of nymphs and adults of A. auricularium on cats in a new area of occurrence in the semi-arid region of Northeastern Brazil and reports for the first time the presence of anti-Ricketsia antibodies in cats in the region, with R. amblyommatis as the probable infectious agent.