Pathogens

11 pages, 1324 KiB

Open AccessArticle

Experimental Infection of Mice and Ticks with the Human Isolate of Anaplasma phagocytophilum NY-18

by Veronika Urbanová, Eliška Kalinová, Petr Kopáček and Radek Šíma

Pathogens 2022, 11(7), 820; https://doi.org/10.3390/pathogens11070820 - 21 Jul 2022

Cited by 2 | Viewed by 2261

Anaplasma phagocytophilum is the causative agent of tick-borne fever (TBF) and human granulocytic anaplasmosis (HGA) and is currently considered an emerging disease in the USA, Europe, and Asia. The increased prevalence of A. phagocytophilum as a human pathogen requires the detailed characterization of [...] Read more.

Anaplasma phagocytophilum is the causative agent of tick-borne fever (TBF) and human granulocytic anaplasmosis (HGA) and is currently considered an emerging disease in the USA, Europe, and Asia. The increased prevalence of A. phagocytophilum as a human pathogen requires the detailed characterization of human isolates and the implementation of appropriate animal models. In this study, we demonstrated that the dynamics of infection with the human isolate of A. phagocytophilum NY-18 was variable in three different strains of mice (SCID, C3H/HeN, BALB/c). We further evaluated the ability of Ixodes ricinus to acquire and transmit A. phagocytophilum NY-18 and compared it with Ixodes scapularis. Larvae of both tick species effectively acquired the pathogen while feeding on infected mice. The infection rates then decreased during the development to nymphs. Interestingly, molted I. ricinus nymphs were unable to transmit the pathogen to naïve mice, which contrasted with I. scapularis. The results of our study suggest that I. ricinus is not a competent vector for the American human Anaplasma isolate. Further studies are needed to establish reliable transmission models for I. ricinus and European human isolate(s) of A. phagocytophilum. Full article

(This article belongs to the Special Issue Ixodes ricinus and Disease Transmission)

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16 pages, 699 KiB

Open AccessReview

The Current Knowledge on Clostridioides difficile Infection in Patients with Inflammatory Bowel Diseases

by Alina Boeriu, Adina Roman, Crina Fofiu and Daniela Dobru

Pathogens 2022, 11(7), 819; https://doi.org/10.3390/pathogens11070819 - 21 Jul 2022

Cited by 15 | Viewed by 3854

Abstract

Clostridioides difficile (C. difficile) represents a major health burden with substantial economic and clinical impact. Patients with inflammatory bowel diseases (IBD) were identified as a risk category for Clostridioides difficile infection (CDI). In addition to traditional risk factors for C. difficile [...] Read more.

Clostridioides difficile (C. difficile) represents a major health burden with substantial economic and clinical impact. Patients with inflammatory bowel diseases (IBD) were identified as a risk category for Clostridioides difficile infection (CDI). In addition to traditional risk factors for C. difficile acquisition, IBD-specific risk factors such as immunosuppression, severity and extension of the inflammatory disease were identified. C. difficile virulence factors, represented by both toxins A and B, induce the damage of the intestinal mucosa and vascular changes, and promote the inflammatory host response. Given the potential life-threatening complications, early diagnostic and therapeutic interventions are required. The screening for CDI is recommended in IBD exacerbations, and the diagnostic algorithm consists of clinical evaluation, enzyme immunoassays (EIAs) or nucleic acid amplification tests (NAATs). An increased length of hospitalization, increased colectomy rate and mortality are the consequences of concurrent CDI in IBD patients. Selection of CD strains of higher virulence, antibiotic resistance, and the increasing rate of recurrent infections make the management of CDI in IBD more challenging. An individualized therapeutic approach is recommended to control CDI as well as IBD flare. Novel therapeutic strategies have been developed in recent years in order to manage severe, refractory or recurrent CDI. In this article, we aim to review the current evidence in the field of CDI in patients with underlying IBD, pointing to pathogenic mechanisms, risk factors for infection, diagnostic steps, clinical impact and outcomes, and specific management. Full article

(This article belongs to the Special Issue Gastrointestinal Pathogens in Inflammatory Bowel Disease)

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17 pages, 3790 KiB

Open AccessReview

SUMOylation and Viral Infections of the Brain

by Fergan Imbert, Gabrielle Leavitt and Dianne Langford

Pathogens 2022, 11(7), 818; https://doi.org/10.3390/pathogens11070818 - 21 Jul 2022

Cited by 7 | Viewed by 3904

Abstract

The small ubiquitin-like modifier (SUMO) system regulates numerous biological processes, including protein localization, stability and/or activity, transcription, and DNA repair. SUMO also plays critical roles in innate immunity and antiviral defense by mediating interferon (IFN) synthesis and signaling, as well as the expression [...] Read more.

The small ubiquitin-like modifier (SUMO) system regulates numerous biological processes, including protein localization, stability and/or activity, transcription, and DNA repair. SUMO also plays critical roles in innate immunity and antiviral defense by mediating interferon (IFN) synthesis and signaling, as well as the expression and function of IFN-stimulated gene products. Viruses including human immunodeficiency virus-1, Zika virus, herpesviruses, and coronaviruses have evolved to exploit the host SUMOylation system to counteract the antiviral activities of SUMO proteins and to modify their own proteins for viral persistence and pathogenesis. Understanding the exploitation of SUMO is necessary for the development of effective antiviral therapies. This review summarizes the interplay between viruses and the host SUMOylation system, with a special emphasis on viruses with neuro-invasive properties that have pathogenic consequences on the central nervous system. Full article

(This article belongs to the Special Issue Viral Infections and Their Effects on the Central Nervous System (CNS))

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SUMOylation: the Small Ubiquitin-related Modifier (SUMO) pathway. SUMO proteins are processed by a SUMO-specific protease at the C-terminal tail to expose a diglycine (-GG) motif, resulting in a mature SUMO peptide. SUMO is subsequently activated in an ATP-dependent reaction, creating an intermediate thioester bond between the active site of SUMO and the heterodimeric E1-activating enzyme (SAE1/SAE2). Following activation, SUMO is transferred from the E1 enzyme to Ubc9, and finally attached to a target lysine in the protein substrate, which is usually located within the consensus site. This final step is mediated by E3 ligase enzymes that function in substrate recognition and specificity. SUMOylation is reversible (deconjugation), and the same SUMO proteases involved in SUMO maturation catalyze the removal of SUMO from the target substrate. Cross-talk exists between SUMOylation and ubiquitylation through SUMO-targeted ubiquitin ligases (STUbLs). STUbLs are enzymes that catalyze the addition of ubiquitin to proteins that have been previously SUMOylated with SUMO chains. STUbL activity results in target proteins that are modified by both SUMO and ubiquitin, which can be targeted to the proteasome for degradation. AMP: adenosine monophosphate; PPi: pyrophosphate; Ub: ubiquitin. Adapted with permission from [<a href="#B26-pathogens-11-00818" class="html-bibr">26</a>] 2021, Springer Nature. Figure was created with Biorender. Full article ">Figure 2
CTIP2 promotes the establishment of HIV-1 latency in microglia. CTIP2 participates in the establishment of HIV-1 latency in microglia by recruiting a chromatin modifying complex (or viral latency complex) to the viral promoter (HIV-1 LTR). This complex consists of: Sp1, which anchors CTIP2 to the viral promoter and acts as a scaffold for the recruitment of chromatin modifying proteins; HDAC1 and HDAC2, which are responsible for deacetylation of Nuc-1, one of the nucleosomes positioned immediately downstream of the transcriptional start site of the HIV-1 LTR; and the histone methyltransferase Suv39H1, which contributes to HIV-1 silencing through methylation of Nuc-1. CTIP2 also recruits the demethylase, LSD1, and the SUMO E3 ligase, TRIM28, which, in association with CTIP2, contributes to HIV-1 gene silencing. Several of the CTIP2-associated proteins in the viral latency complex interact with the host SUMOylation system. Accordingly, determining the role of the SUMOylation in the establishment and/or persistence of HIV-1 latency in microglia could aid in the design of new pharmacological agents that target HIV-1 viral reservoirs. Sp1: specificity protein 1; COUP-TF: chicken ovalbumin upstream promoter transcription factor; CTIP2: COUP-TF interacting protein 2; HDAC1/2: histone deacetylase 1/2; TRIM28: tripartite motif containing 28; SUMO: small ubiquitin-related modifier. Figure was created with Biorender. Full article ">Figure 3
Inflammatory pathology in COVID-19 brains. (A) Section of the hypoglossal nucleus shows several motor neurons and a microglial nodule (arrow). (B) An adjacent section immunolabeled for CD68 (brown) shows clustered microglia within the nodule. Inset: Microglia in close apposition to a hypoglossal neuron (CD68+). (E) The locus coeruleus contains a microglial nodule with a degenerating neuron in the center, identified by its residual neuromelanin (arrow). (F,G) Neurons of the dorsal motor nucleus of the vagus surrounded by CD68+ microglia. (H,I) Microglial nodules in the dentate nucleus (arrows in (H)), neuron in the middle of a nodule (arrow in (I)), CD68. Scale bar in (A,B) = 200 µm; (E) = 10 µm; (F,G) = 50 µm; (H) = 100 µm; (I) = 50 µm. Adapted with permission from [<a href="#B61-pathogens-11-00818" class="html-bibr">61</a>] 2021, Oxford University Press. Panels C, D, J, K from the original publication are not shown. Full article ">Figure 4
ZIKV NS5 forms unique nuclear speckles that disrupt PML/SUMO-1 NBs. (A) hBMECs grown on microslides were infected with ZIKV-PRV (MOI, 2) and fixed 12, 18, and 24 hpi; immunolabeled for ZIKV NS5 and SUMO-1 or PML; and visualized by confocal microscopy. (B) Mock-infected or ZIKV-infected hBMECs were immunolabeled for PML and SUMO-1 24 hpi and visualized by confocal microscopy. (C) ZIKV-infected hBMECs were immunolabeled for ZIKV NS5 and fibrillarin at 24 hpi and visualized by confocal microscopy. Experiments were conducted in triplicate, repeated at least three times, and representative data are presented. Bars = 5 μm. Adapted with permission from [<a href="#B76-pathogens-11-00818" class="html-bibr">76</a>]. 2020, American Society for Microbiology. PML: promyelocytic leukemia; ZIKV-PRV: PRVABC59 ZIKV strain; hBMEC: human brain microvascular endothelial cells; MOI: multiplicity of infection; hpi: hours post-infection. Full article ">Figure 5
Molecular docking model of the binding between the putative non-structural 5 (NS5) SUMO-interacting motifs of flaviviruses and the SUMO1 protein. (A) Top panel: multiple sequence alignment of the amino acid sequences of the putative NS5 protein SUMO-interacting motifs (SIM) of Zika virus (ZIKV), dengue virus (DENV) (serotype 3), Japanese encephalitis virus (JEV), West Nile virus (WNV), and yellow fever virus (YFV). Bottom panel: Stick representation of the structural similarities among the five flaviviruses’ putative NS5 SIM peptides. (B) Schematic representation of the binding between the SUMO1 protein and the five flaviviruses’ putative NS5 SIM peptides. The SUMO1 protein is shown in tan and the NS5 SIM peptides are shown in different colors. (C) Ribbon representation showing the interacting amino acid residues of the putative ZIKV NS5 SIM peptide and the active sites of the SUMO1 protein. The putative ZIKV NS5 SIM peptide and SUMO1 protein are displayed in magenta and blue, respectively. The interacting residues are shown as sticks with hydrogen bonds represented by yellow dashed lines. Adapted with permission from [<a href="#B74-pathogens-11-00818" class="html-bibr">74</a>]. 2019, MDPI. Full article ">Figure 6
Interactions between human cytomegalovirus (HCMV) and SUMO. HCMV entry to the central nervous system (CNS) is secondary to peripheral organ infection. Passage across the blood-brain barrier is thought to be mediated by monocytes. Upon crossing the BBB, HCMV infects resident cells and has been shown to interact with the host SUMOylation system. Immediate–early protein-1 (IE1) was the first viral protein identified as a SUMO interactor and has been shown to inhibit the SUMOylation of promyelocytic leukemia (PML) bodies, which suppresses innate immune responses. Similarly, the HCMV latency-associated protein, LUNA, functions as a de-SUMOylase to promote PML de-SUMOylation. The HCMV pp71 protein promotes the SUMOylation of its cellular substrate, Daxx, though the functional consequence of this interaction is unknown. pp71 has also been shown to mediate Daxx degradation through a ubiquitin-independent pathway. Figure was created with Biorender. Full article ">

12 pages, 1923 KiB

Open AccessArticle

Variability among the Isolates of Broad Bean Mottle Virus and Encapsidation of Host RNAs

by Nipin Shrestha, Melvin R. Duvall and Jozef J. Bujarski

Pathogens 2022, 11(7), 817; https://doi.org/10.3390/pathogens11070817 - 21 Jul 2022

Cited by 1 | Viewed by 2003

Abstract

Broad bean mottle bromovirus infects legume plants and is transmissible by insects. Several broad bean mottle virus (BBMV) isolates have been identified, including one in England (isolate Ba) and five in the Mediterranean countries: Libya (LyV), Morocco (MV), Syria (SV), Sudan (TU) and [...] Read more.

Broad bean mottle bromovirus infects legume plants and is transmissible by insects. Several broad bean mottle virus (BBMV) isolates have been identified, including one in England (isolate Ba) and five in the Mediterranean countries: Libya (LyV), Morocco (MV), Syria (SV), Sudan (TU) and Tunisia (TV). Previously, we analyzed the nucleotide sequence of the Ba RNA and here we report on and compare it with another five Mediterranean variants. The RNA segments in the latter ones were extensively homologous, with some SNPs, single nucleotide deletions and insertions, while the number of mutations was higher in isolate Ba. Both the 5′ and 3′ untranslated terminal regions (UTRs) among the corresponding RNAs are highly conserved, reflecting their functionality in virus replication. The AUG initiation codons are within suboptimal contexts, possibly to adjust/regulate translation. The proteins 1a, 2a, 3a and coat protein (CP) are almost identical among the five isolates, but in Ba they have more amino acid (aa) substitutions. Phylogenetic analysis revealed the isolates from Morocco and Syria clustering with the isolate from England, while the variants from Libya, Tunisia and Sudan created a different clade. The BBMV isolates encapsidate a high content of host (ribosomal and messenger) RNAs. Our studies present BBMV as a useful model for bromoviruses infecting legumes. Full article

(This article belongs to the Special Issue 10th Anniversary of Pathogens—Feature Papers)

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Denaturing gel of the BBMV RNAs isolated from five BBMV isolates. The images were taken prior to sequencing. L1, L2, L3, L4 and L5, which are the lanes loaded with the RNA from MV, LyV, SV, TU and TV isolates, respectively. Lane M is the size marker. The three major bands, characteristic for Bromoviruses, representing RNA1 + 2 (~2900–3100 bp), RNA3 (~2200 bp) and sgRNA4 (~800 bp) are visible in the gel (indicated on the right side). Full article ">Figure 2
A stacked plot showing proportion of reads that mapped to RNA1, RNA2 and RNA3 among the five BBMV isolates. Full article ">Figure 3
Multiple sequence alignment (MSA) of nucleotide sequences of BBMV RNAs 1, 2, and 3 (top to bottom) for five BBMV isolates as compared to Ba ones. The orange bars represent the ORF regions (with the length shown in parentheses). The gray regions signify identical nucleotides, while the nucleotide substitutions are depicted by the black vertical lines and the deletions by the horizontal black lines. The green bars illustrate the regions of identity. Full article ">Figure 4
The alignment of the nt sequences of the intercistronic noncoding region in the RNA3 of the BBMV variants covering the subgenomic promoter core (black bar) and the start site of the sg RNA4 transcription (see also Romero et al., 1992 [<a href="#B6-pathogens-11-00817" class="html-bibr">6</a>]). The subgenomic RNA start site are marked by red bar, and the CP translation start codon ATG is indicated by the blue bar on top of the alignment. Full article ">Figure 5
MSA of amino acid sequences of proteins 1a, 2a, 3a and 4a (CP) for six BBMV isolates. The gray regions represent identical nucleotides, while the aa substitutions are depicted by the black vertical lines and the deletions by small gaps. The green bars illustrate the regions of identity. The numerical scale on top marks the positions of amino acids. Full article ">Figure 6
Maximum likelihood (ML) bootstrap consensus cladogram for five Mediterranean and the England isolates of BBMV. The sequences of RNAs 1, 2 and 3 were concatenated and aligned by MSA followed by the consensus tree reconstruction by the ML bootstrap method. The BMV RNA sequences were used as the outgroup to infer the rooted cladogram. Numbers along the branches are ML bootstrap values. The cladogram has been calculated at the Geneious default values. Full article ">Figure 7
The stacked plot illustration profiling the proportions of different types of encapsidated host RNAs in BBMV vs. BMV, originating from three different cellular compartments. Apparently in BBMV, more than 99% of the host RNAs were nuclear RNAs with ribosomal RNAs being present more in comparison to the mRNA transcripts, whereas in BMV over 50% of the reads mapped to the nuclear RNAs followed by the organellar RNAs (mitochondrial and plastid). The proportion of the mRNAs being mapped dominantly in comparison to the rRNAs also differed between BBMV and BMV. Full article ">

10 pages, 754 KiB

Open AccessReview

COVID-19 as Another Trigger for HBV Reactivation: Clinical Case and Review of Literature

by Caterina Sagnelli, Laura Montella, Pierantonio Grimaldi, Mariantonietta Pisaturo, Loredana Alessio, Stefania De Pascalis, Evangelista Sagnelli and Nicola Coppola

Pathogens 2022, 11(7), 816; https://doi.org/10.3390/pathogens11070816 - 21 Jul 2022

Cited by 20 | Viewed by 3319

Abstract

Universal hepatitis B virus (HBV) vaccination has been applied for years in most countries, but HBV infection remains an unresolved public health problem worldwide, with over one-third of the world’s population infected during their lifetime and approximately 248 million hepatitis B surface antigen [...] Read more.

Universal hepatitis B virus (HBV) vaccination has been applied for years in most countries, but HBV infection remains an unresolved public health problem worldwide, with over one-third of the world’s population infected during their lifetime and approximately 248 million hepatitis B surface antigen (HBsAg) chronic carriers. HBV infection may reactivate with symptomatic and sometimes life-threatening clinical manifestations due to a reduction in the immune response of various origins, due to chemotherapy or immunosuppressive therapy, treatments increasingly practiced worldwide. SARS-CoV-2 and its COVID-19 associated disease have introduced new chances for HBV reactivation due to the use of dexamethasone and tocilizumab to counteract the cytokine storm. This could and should be prevented by accurate screening of HBV serologic markers and adequate pharmacologic prophylaxis. This article describes the case of a patient with COVID-19 who developed HBV reactivation and died of liver failure and analyzes published data on this setting to provide useful information to physicians who manage these patients during the SARS-CoV-2 pandemic. Full article

(This article belongs to the Special Issue Viral Hepatitis: The New Challenge in the Era of Antiviral Treatments)

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9 pages, 2471 KiB

Open AccessCase Report

A Ruptured Left Gastric Artery Aneurysm That Neoplasticized during the Course of Coronavirus Disease 2019: A Case Report

by Satoshi Ano, Yuto Shinkura, Tsuneaki Kenzaka, Naoaki Kusunoki, Satoru Kawasaki and Hogara Nishisaki

Pathogens 2022, 11(7), 815; https://doi.org/10.3390/pathogens11070815 - 20 Jul 2022

Cited by 3 | Viewed by 2185

Abstract

Coronavirus disease 2019 (COVID-19) is an acute respiratory syndrome caused by SARS-CoV-2 and is known to cause respiratory and systemic symptoms. A SARS-CoV-2 infection is involved in aneurysm formation, enlargement, and rupture in medium-sized vessels, such as the cerebral and coronary arteries and [...] Read more.

Coronavirus disease 2019 (COVID-19) is an acute respiratory syndrome caused by SARS-CoV-2 and is known to cause respiratory and systemic symptoms. A SARS-CoV-2 infection is involved in aneurysm formation, enlargement, and rupture in medium-sized vessels, such as the cerebral and coronary arteries and the aorta. In contrast, its involvement in forming aneurysms in medium-sized vessels other than the cerebral and coronary arteries has not been reported. An 84-year-old Japanese man with COVID-19 was admitted to our hospital. The treatment course was favorable, and the COVID-19 treatment was completed by the 10th day. On day 14, pancreatic enzymes increased mildly. An abdominal computed tomography revealed a ruptured left gastric aneurysm after spontaneous hemostasis. Arterial embolization was performed. In this patient, a new left gastric aneurysm was suspected of having formed and ruptured during the course of the COVID-19 treatment. To the best of our knowledge, this is the first report of abdominal visceral aneurysm formation caused by COVID-19 in a medium-sized vessel, and it is necessary to remember that aneurysms can be formed at any site when treating this syndrome. Full article

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Seated frontal view of the chest radiograph during admission. The whole lung fields on both sides are observed in frosted shadows (yellow arrows). Full article ">Figure 2
Non-contrast computed tomography of the chest at admission. Diffuse frosted shadows in both lungs and an infiltrative shadow in the right lower lobe can be observed (yellow arrows). Full article ">Figure 3
Post-hospitalization course. Full article ">Figure 4
(a) Non-contrast computed tomography of the abdomen on admission. Gallstones were found in the neck region of the gallbladder. However, there were no significant findings in the dorsal gastric or ventral pancreas. (b) Non-contrast computed tomography of the abdomen on day 14. A highly absorptive zone in the dorsal gastric/peripancreatic region (yellow arrows) and dilatation of the left gastric artery (red arrows) were observed. Full article ">Figure 5
Abdominal contrast computed tomography (Maximum Intensity Projection image) on day 16. A bead-shaped aneurysm formation was seen in the left gastric artery (yellow arrows). The patient was diagnosed after rupture and spontaneous hemostasis of the left gastric artery aneurysm. Full article ">Figure 6
(a) Abdominal angiography before arterial embolization. The left gastric artery shows a bead-shaped dilatation (yellow arrows). (b) Post-arterial embolization abdominal angiography. Blood flow to the aneurysmal lesion was interrupted (yellow arrows). Full article ">

9 pages, 2700 KiB

Open AccessArticle

The Biological Properties of the SARS-CoV-2 Cameroon Variant Spike: An Intermediate between the Alpha and Delta Variants

by Stefano Pascarella, Martina Bianchi, Marta Giovanetti, Domenico Benvenuto, Alessandra Borsetti, Roberto Cauda, Antonio Cassone and Massimo Ciccozzi

Pathogens 2022, 11(7), 814; https://doi.org/10.3390/pathogens11070814 - 20 Jul 2022

Cited by 1 | Viewed by 1938

Abstract

An analysis of the structural effect of the mutations of the B.1.640.2 (IHU) Spike Receptor Binding Domain (RBD) and N-terminal Domain (NTD) is reported along with a comparison with the sister lineage B.1.640.1. and a selection of variants of concern. The effect of [...] Read more.

An analysis of the structural effect of the mutations of the B.1.640.2 (IHU) Spike Receptor Binding Domain (RBD) and N-terminal Domain (NTD) is reported along with a comparison with the sister lineage B.1.640.1. and a selection of variants of concern. The effect of the mutations on the RBD–ACE2 interaction was also assessed. The structural analysis applied computational methods that are able to carry out in silico mutagenesis to calculate energy minimization and the folding energy variation consequent to residue mutations. Tools for electrostatic calculation were applied to quantify and display the protein surface electrostatic potential. Interactions at the RBD–ACE2 interface were scrutinized using computational tools that identify the interactions and predict the contribution of each interface residue to the stability of the complex. The comparison among the RBDs shows that the most evident differences between the variants is in the distribution of the surface electrostatic potential: that of B.1.640.1 is as that of the Alpha RBD, while B.1.640.2 appears to have an intermediate surface potential pattern with characteristics between those of the Alpha and Delta variants. Moreover, the B.1.640.2 Spike includes the mutation E484K that in other variants has been suggested to be involved in immune evasion. These properties may hint at the possibility that B.1.640.2 emerged with a potentially increased infectivity with respect to the sister B.1.640.1 variant, but significantly lower than that of the Delta and Omicron variants. However, the analysis of their NTD domains highlights deletions, destabilizing mutations and charge alterations that can limit the ability of the B.1.640.1 and B.1.640.2 variants to interact with cellular components, such as cell surface receptors. Full article

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9 pages, 262 KiB

Open AccessArticle

A Retrospective Study of the Efficacy of Albendazole and Diethylcarbamazine for the Treatment of Human Toxocariasis

by Jean-François Magnaval, Judith Fillaux and Antoine Berry

Pathogens 2022, 11(7), 813; https://doi.org/10.3390/pathogens11070813 - 20 Jul 2022

Cited by 6 | Viewed by 2372

Abstract

In the Department of Parasitology and Mycology of Toulouse University Hospitals, patients presenting with common/covert toxocariasis were treated either with albendazole (39 cases) or with diethylcarbamazine (32 cases). Albendazole (ABZ) was given at 10 mg/kg b/w daily for 14 days, and diethylcarbamazine (DEC) [...] Read more.

In the Department of Parasitology and Mycology of Toulouse University Hospitals, patients presenting with common/covert toxocariasis were treated either with albendazole (39 cases) or with diethylcarbamazine (32 cases). Albendazole (ABZ) was given at 10 mg/kg b/w daily for 14 days, and diethylcarbamazine (DEC) was given at 4 mg/kg b/w daily for 21 days. In both groups, follow-up consultations occurred approximately 48 days after the end of the anthelmintic therapy. ABZ and DEC displayed a similar efficacy on the kinetics of the clinical picture (−64.5% of reduction vs. −72.7%, respectively) and on the levels of blood eosinophilia, serum eosinophil cationic protein and serum total IgE. However, the effect of the medication on the laboratory parameters was moderate. The rate of adverse reactions was similar in both groups (38% for ABZ vs. 31% for DEC), but DEC-treated patients complained of more intense and long-lasting side effects. The DEC group had more major adverse reactions, resulting in the termination of the anthelmintic treatment. The results from this retrospective study bring further arguments for considering ABZ, given at 10 mg/kg daily for 2 weeks, as the drug of choice in the treatment of human toxocariasis. Full article

(This article belongs to the Special Issue Bacterial, Fungal and Parasitic Zoonoses)

12 pages, 1676 KiB

Open AccessArticle

Survival of Campylobacter jejuni Co-Cultured with Salmonella spp. in Aerobic Conditions

by Nagham Anis, Laetitia Bonifait, Ségolène Quesne, Louise Baugé, Wissam Yassine, Muriel Guyard-Nicodème and Marianne Chemaly

Pathogens 2022, 11(7), 812; https://doi.org/10.3390/pathogens11070812 - 20 Jul 2022

Cited by 4 | Viewed by 2583

Abstract

Campylobacter and Salmonella are responsible for the two major foodborne zoonotic diseases in Europe; poultry is the main infection source. Campylobacter cannot grow under aerobic conditions, but can show aerobic survival when co-cultured with other microorganisms; however, its interaction with Salmonella has not [...] Read more.

Campylobacter and Salmonella are responsible for the two major foodborne zoonotic diseases in Europe; poultry is the main infection source. Campylobacter cannot grow under aerobic conditions, but can show aerobic survival when co-cultured with other microorganisms; however, its interaction with Salmonella has not been studied yet. In this study, these two bacteria were co-cultured under controlled aerobic conditions. Different concentrations and strains of C. jejuni were incubated with or without different Salmonella serotypes (10 CFU) at 37 °C for 16 h. C. jejuni did not grow after incubation with or without Salmonella. The survival of C. jejuni was observed only for the highest initial concentration of 6 log CFU/mL with or without Salmonella. However, its survival was significantly higher when co-cultured with Salmonella. No survival was observed at lower concentrations. C. jejuni survival was positively affected by the presence of Salmonella but depended on the Salmonella serotype, the C. jejuni strain and the initial concentration. On the other hand, the Salmonella enumerations were not affected by C. jejuni. Our results suggest potential interactions between Salmonella and C. jejuni that require further investigations for a clearer understanding of their behavior in natural habitats. Full article

(This article belongs to the Collection Campylobacter Infections Collection)

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C. jejuni C97Anses640 counts before and after incubation under aerobic conditions (with or without Salmonella Blegdam) (n = 10: (a–c) from 6 to 4 log CFU/mL, n = 4: (d,e) for 3 and 2 log CFU/mL). C bf inc: Initial concentration of C97Anses640 before incubation; C + S bf inc: Initial concentration of C97Anses640 co-cultured with S. Blegdam before incubation; C af inc: Final concentration of C97Anses640 after aerobic incubation; C + S af inc: Final concentration of C97Anses640 co-cultured with S. Blegdam after aerobic incubation. Full article ">Figure 2
C. jejuni C97Anses640 counts before and after incubation under aerobic conditions. (a) With or without Salmonella Typhimurium; (b) With or without Salmonella Enteritidis (n = 1). Full article ">Figure 3
C. jejuni strain counts before and after incubation of 4 log CFU/mL under aerobic conditions with or without different Salmonella serovars (light blue bars: initial concentration of C. jejuni; light gray bars: initial concentration of C. jejuni co-cultured with Salmonella; dark blue bars: final concentration of C. jejuni following aerobic incubation; dark gray bars: final concentration of C. jejuni co-cultured with Salmonella following aerobic incubation). (a) Six strains of C. jejuni were co-cultured with or without S. Blegdam 421, S. Typhimurium (S17 LNR1383) and S. Enteritidis (S17 LNR01420); (b) C. jejuni strains AC 302 and AC 541 co-cultured with or without S. Typhimurium (S20 LNR0260) and S. Enteritidis (S20 LNR0176) compared to S. Typhimurium (S17 LNR1383) and S. Enteritidis (S17 LNR01420). Full article ">Figure 4
Experimental protocol for survival assays: A culture of C. jejuni (from 8 to 4 log CFU/mL) was diluted to inoculate the bags, containing 250 mL buffered peptone water, with different final concentrations of C. jejuni ranging from 6 to 2 log CFU/mL. Then, C. jejuni obtained in the bags were cultured under aerobic conditions at 37 °C for 16 h in the presence or absence of 10 CFU of Salmonella spp. Full article ">

5 pages, 208 KiB

Open AccessEditorial

Advances in the Immunobiology of Parasitic Diseases

by Jorge Morales-Montor, Derek M. McKay and Luis I. Terrazas

Pathogens 2022, 11(7), 811; https://doi.org/10.3390/pathogens11070811 - 20 Jul 2022

Viewed by 1846

Abstract

Notwithstanding that most biomedical research today focuses on the pandemic caused by the SARs-CoV-2 virus, there are many unresolved diseases that are almost forgotten worldwide [...] Full article

(This article belongs to the Special Issue Advances in the Immunobiology of Parasitic Diseases)

13 pages, 1939 KiB

Open AccessArticle

Assembly, Annotation, and Comparative Whole Genome Sequence of Fusarium verticillioides Isolated from Stored Maize Grains

by Vishwambar D. Navale, Amol M. Sawant, Varun U. Gowda and Koteswara Rao Vamkudoth

Pathogens 2022, 11(7), 810; https://doi.org/10.3390/pathogens11070810 - 20 Jul 2022

Cited by 4 | Viewed by 3101

Abstract

Fusarium verticillioides is a plant pathogenic fungus affecting a wide range of crops worldwide due to its toxigenic properties. F. verticillioides BIONCL4 strain was isolated from stored maize grain samples in India, and produces high amount of fumonisin B1 (FB1). We report a [...] Read more.

Fusarium verticillioides is a plant pathogenic fungus affecting a wide range of crops worldwide due to its toxigenic properties. F. verticillioides BIONCL4 strain was isolated from stored maize grain samples in India, and produces high amount of fumonisin B1 (FB1). We report a comparative genomic analysis of F. verticillioides, covering the basic genome information, secretome, and proteins involved in host–pathogen interactions and mycotoxin biosynthesis. Whole-genome sequencing (WGS) was performed using the Illumina platform with an assembly size of 42.91 Mb, GC content of 48.24%, and 98.50% coverage with the reference genome (GCA000149555). It encodes 15,053 proteins, including 2058 secretory proteins, 676 classical secretory proteins, and 569 virulence and pathogenicity-related proteins. There were also 1447 genes linked to carbohydrate active enzymes (CaZymes) and 167 genes related to mycotoxin production. Furthermore, F. verticillioides genome comparison revealed information about the species’ evolutionary history. The overall study helps in disease prevention and management of mycotoxins to ensure food safety. Full article

(This article belongs to the Special Issue Mycotoxigenic Fungi Associated with Food Commodities during Cultivation, Storage and Marketing)

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14 pages, 1571 KiB

Open AccessReview

Role of Microglia in Herpesvirus-Related Neuroinflammation and Neurodegeneration

by Magdalena Patrycy, Marcin Chodkowski and Malgorzata Krzyzowska

Pathogens 2022, 11(7), 809; https://doi.org/10.3390/pathogens11070809 - 19 Jul 2022

Cited by 18 | Viewed by 3611

Abstract

Neuroinflammation is defined as an inflammatory state within the central nervous system (CNS). Microglia conprise the resident tissue macrophages of the neuronal tissue. Upon viral infection of the CNS, microglia become activated and start to produce inflammatory mediators important for clearance of the [...] Read more.

Neuroinflammation is defined as an inflammatory state within the central nervous system (CNS). Microglia conprise the resident tissue macrophages of the neuronal tissue. Upon viral infection of the CNS, microglia become activated and start to produce inflammatory mediators important for clearance of the virus, but an excessive neuroinflammation can harm nearby neuronal cells. Herpesviruses express several molecular mechanisms, which can modulate apoptosis of infected neurons, astrocytes and microglia but also divert immune response initiated by the infected cells. In this review we also describe the link between virus-related neuroinflammation, and development of neurodegenerative diseases. Full article

(This article belongs to the Special Issue Modification of Cellular Response by HSV)

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23 pages, 1784 KiB

Open AccessArticle

Molecular Epidemiology and Baseline Resistance of Hepatitis C Virus to Direct Acting Antivirals in Croatia

by Petra Simicic, Anamarija Slovic, Leona Radmanic, Adriana Vince and Snjezana Zidovec Lepej

Pathogens 2022, 11(7), 808; https://doi.org/10.3390/pathogens11070808 - 19 Jul 2022

Viewed by 2079

Abstract

Molecular epidemiology of hepatitis C virus (HCV) is exceptionally complex due to the highly diverse HCV genome. Genetic diversity, transmission dynamics, and epidemic history of the most common HCV genotypes were inferred by population sequencing of the HCV NS3, NS5A, and NS5B region [...] Read more.

Molecular epidemiology of hepatitis C virus (HCV) is exceptionally complex due to the highly diverse HCV genome. Genetic diversity, transmission dynamics, and epidemic history of the most common HCV genotypes were inferred by population sequencing of the HCV NS3, NS5A, and NS5B region followed by phylogenetic and phylodynamic analysis. The results of this research suggest high overall prevalence of baseline NS3 resistance associate substitutions (RAS) (33.0%), moderate prevalence of NS5A RAS (13.7%), and low prevalence of nucleoside inhibitor NS5B RAS (8.3%). Prevalence of RAS significantly differed according to HCV genotype, with the highest prevalence of baseline resistance to NS3 inhibitors and NS5A inhibitors observed in HCV subtype 1a (68.8%) and subtype 1b (21.3%), respectively. Phylogenetic tree reconstructions showed two distinct clades within the subtype 1a, clade I (62.4%) and clade II (37.6%). NS3 RAS were preferentially associated with clade I. Phylogenetic analysis demonstrated that 27 (9.0%) HCV sequences had a presumed epidemiological link with another sequence and classified into 13 transmission pairs or clusters which were predominantly comprised of subtype 3a viruses and commonly detected among intravenous drug users (IDU). Phylodynamic analyses highlighted an exponential increase in subtype 1a and 3a effective population size in the late 20th century, which is a period associated with an explosive increase in the number of IDU in Croatia. Full article

(This article belongs to the Special Issue Molecular Diagnostic and Epidemiology of Viral Infections)

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Prevalence of overall and resistance conferring RAS according to geno2pheno [HCV] algorithm and HCV genotype in: (a) NS3 region (b) NS5A region. Full article ">Figure 2
Maximum likelihood phylogenetic analysis of the HCV NS5B gene sequences constructed by applying GTR+G+I model with 1000 bootstrap replicates. Scale bar represents 0.1 nucleotide substitutions per site. Bootstrap values between 70 and 100% are displayed at the branch nodes as blue triangles with circle size corresponding to magnitude of bootstrap. Branches of reference sequences are colored black, while branches of Croatian sequences are colored red. Genotypes and subtypes are indicated by different color strips. Full article ">Figure 3
Maximum likelihood phylogenetic analysis of the NS3 gene sequences of HCV 1a subtype constructed with GTR+G+I model and 1000 bootstrap replicates. Bootstrap values between 70 and 100% are displayed at the branch nodes as blue triangles with size corresponding to magnitude of bootstrap. Branches of two most similar control sequences per each local sequence obtained by searching the BLAST database and removing duplicates are colored black. Branches of Croatian sequences without RAS are colored green, sequences with RAS conferring resistance to DAA are colored red, and sequences with RAS associated with reduced susceptibility to DAA are colored orange. Reference sequences are colored gray. Clade I sequences are highlighted blue, while clade II sequences are highlighted pink. All identified RAS are positioned on the phylogenetic tree along with the corresponding sequences. Resistance conferring RAS are marked with filled symbols, while RAS causing reduced susceptibility to at least one DAA are marked with open symbols. Identified transmission pairs are highlighted gray, while transmission pairs identified consistently across all genomic regions for GT1a are highlighted yellow. Full article ">Figure 4
Bayesian skyline plots showing the epidemic history of the HCV subtype 1a, 1b, and 3a sequences in Zagreb, Croatia. Mean (solid blue line) and upper and lower 95% HPD (solid blue area) estimates of the effective population size (Y-axis; log10 scale) through time (X-axis; calendar years) from the time of the most recent common ancestor (tMRCA) are shown. Full article ">

16 pages, 1558 KiB

Open AccessArticle

Disturbance Ecology Meets Bovine Tuberculosis (bTB) Epidemiology: A Before-and-After Study on the Association between Forest Clearfelling and bTB Herd Risk in Cattle Herds

by Andrew W. Byrne, Damien Barrett, Philip Breslin, James O’Keeffe, Kilian J. Murphy, Kimberly Conteddu, Virginia Morera-Pujol, Eoin Ryan and Simone Ciuti

Pathogens 2022, 11(7), 807; https://doi.org/10.3390/pathogens11070807 - 19 Jul 2022

Cited by 5 | Viewed by 3008

Abstract

Disturbance ecology refers to the study of discrete processes that disrupt the structure or dynamics of an ecosystem. Such processes can, therefore, affect wildlife species ecology, including those that are important pathogen hosts. We report on an observational before-and-after study on the association [...] Read more.

Disturbance ecology refers to the study of discrete processes that disrupt the structure or dynamics of an ecosystem. Such processes can, therefore, affect wildlife species ecology, including those that are important pathogen hosts. We report on an observational before-and-after study on the association between forest clearfelling and bovine tuberculosis (bTB) herd risk in cattle herds, an episystem where badgers (Meles meles) are the primary wildlife spillover host. The study design compared herd bTB breakdown risk for a period of 1 year prior to and after exposure to clearfelling across Ireland at sites cut in 2015–2017. The percent of herds positive rose from 3.47% prior to clearfelling to 4.08% after exposure. After controlling for confounders (e.g., herd size, herd type), we found that cattle herds significantly increased their odds of experiencing a bTB breakdown by 1.2-times (95%CIs: 1.07–1.36) up to 1 year after a clearfell risk period. Disturbance ecology of wildlife reservoirs is an understudied area with regards to shared endemic pathogens. Epidemiological observational studies are the first step in building an evidence base to assess the impact of such disturbance events; however, such studies are limited in inferring the mechanism for any changes in risk observed. The current cohort study suggested an association between clearfelling and bTB risk, which we speculate could relate to wildlife disturbance affecting pathogen spillback to cattle, though the study design precludes causal inference. Further studies are required. However, ultimately, integration of epidemiology with wildlife ecology will be important for understanding the underlying mechanisms involved, and to derive suitable effective management proposals, if required. Full article

(This article belongs to the Section Bacterial Pathogens)

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17 pages, 1606 KiB

Open AccessArticle

GMP Manufacturing and IND-Enabling Studies of a Recombinant Hyperimmune Globulin Targeting SARS-CoV-2

by Rena A. Mizrahi, Wendy Y. Lin, Ashley Gras, Ariel R. Niedecken, Ellen K. Wagner, Sheila M. Keating, Nikita Ikon, Vishal A. Manickam, Michael A. Asensio, Jackson Leong, Angelica V. Medina-Cucurella, Emily Benzie, Kyle P. Carter, Yao Chiang, Robert C. Edgar, Renee Leong, Yoong Wearn Lim, Jan Fredrik Simons, Matthew J. Spindler, Kacy Stadtmiller, Nicholas Wayham, Dirk Büscher, Jose Vicente Terencio, Clara Di Germanio, Steven M. Chamow, Charles Olson, Paula A. Pino, Jun-Gyu Park, Amberlee Hicks, Chengjin Ye, Andreu Garcia-Vilanova, Luis Martinez-Sobrido, Jordi B. Torrelles, David S. Johnson and Adam S. Adler Show full author list Hide full author list

Pathogens 2022, 11(7), 806; https://doi.org/10.3390/pathogens11070806 - 19 Jul 2022

Cited by 6 | Viewed by 4541

Abstract

Conventionally, hyperimmune globulin drugs manufactured from pooled immunoglobulins from vaccinated or convalescent donors have been used in treating infections where no treatment is available. This is especially important where multi-epitope neutralization is required to prevent the development of immune-evading viral mutants that can [...] Read more.

Conventionally, hyperimmune globulin drugs manufactured from pooled immunoglobulins from vaccinated or convalescent donors have been used in treating infections where no treatment is available. This is especially important where multi-epitope neutralization is required to prevent the development of immune-evading viral mutants that can emerge upon treatment with monoclonal antibodies. Using microfluidics, flow sorting, and a targeted integration cell line, a first-in-class recombinant hyperimmune globulin therapeutic against SARS-CoV-2 (GIGA-2050) was generated. Using processes similar to conventional monoclonal antibody manufacturing, GIGA-2050, comprising 12,500 antibodies, was scaled-up for clinical manufacturing and multiple development/tox lots were assessed for consistency. Antibody sequence diversity, cell growth, productivity, and product quality were assessed across different manufacturing sites and production scales. GIGA-2050 was purified and tested for good laboratory procedures (GLP) toxicology, pharmacokinetics, and in vivo efficacy against natural SARS-CoV-2 infection in mice. The GIGA-2050 master cell bank was highly stable, producing material at consistent yield and product quality up to >70 generations. Good manufacturing practices (GMP) and development batches of GIGA-2050 showed consistent product quality, impurity clearance, potency, and protection in an in vivo efficacy model. Nonhuman primate toxicology and pharmacokinetics studies suggest that GIGA-2050 is safe and has a half-life similar to other recombinant human IgG1 antibodies. These results supported a successful investigational new drug application for GIGA-2050. This study demonstrates that a new class of drugs, recombinant hyperimmune globulins, can be manufactured consistently at the clinical scale and presents a new approach to treating infectious diseases that targets multiple epitopes of a virus. Full article

(This article belongs to the Collection SARS-CoV-2 Infection and COVID-19 Disease)

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Process flow diagram outlining the upstream and downstream manufacturing process for GIGA-2050, which is similar to a standard mAb process. After fed-batch production in a conventional single-use bioreactor, the process includes clarification via depth filtration, concentration of the clarified harvest followed by Protein A chromatography, low pH viral inactivation, cation exchange chromatography in bind and elute mode, anion exchange chromatography in flow through mode, viral filtration and ultrafiltration/diafiltration to concentrate, and buffer exchange into the final formulation. Full article ">Figure 2
Jaccard and Morisita statistical analyses of antibody RNA-Seq data show a high degree of sequence similarity (>0.95 for Jaccard and >0.98 for Morisita) between two representative development (Dev) lots, the Tox lot, and the GMP lot. Analysis of PCR replicates from these lots found that the lot-to-lot variability was no greater than the variability across PCR replicates from a single lot, indicating strong batch-to-batch consistency. Full article ">Figure 3
Kaplan–Meier curves for K18 hACE2 transgenic mice after SARS-CoV-2 infection. The survival probability is reported for uninfected controls (black), infected but no treatment control (green), reference mAb (CC12.3; 1.5 mg/kg; red), and GIGA-2050 (5 mg/kg; blue). Day after challenge with SARS-CoV-2 is on the x-axis. Animals were treated with GIGA-2050 or the reference mAb 24 h before infection with SARS-CoV-2. For treatment with GIGA-2050, the probability of survival is significantly higher than the no treatment infected control (p = 0.007, Mantel–Cox). The reference mAb at the concentration studied (1.5 mg/kg) did not significantly reduce mortality compared to no treatment control (p = 0.13, Mantel–Cox). Full article ">

10 pages, 807 KiB

Open AccessArticle

Preparation and Storage of Cryoprecipitate Derived from Amotosalen and UVA-Treated Apheresis Plasma and Assessment of In Vitro Quality Parameters

by Katarina Kovacic Krizanic, Florian Prüller, Konrad Rosskopf, Jean-Marc Payrat, Silke Andresen and Peter Schlenke

Pathogens 2022, 11(7), 805; https://doi.org/10.3390/pathogens11070805 - 18 Jul 2022

Cited by 5 | Viewed by 3361

Abstract

Cryoprecipitate is a plasma-derived blood product, enriched for fibrinogen, factor VIII, factor XIII, and von Willebrand factor. Due to infectious risk, the use of cryoprecipitate in Central Europe diminished over the last decades. However, after the introduction of various pathogen-reduction technologies for plasma, [...] Read more.

Cryoprecipitate is a plasma-derived blood product, enriched for fibrinogen, factor VIII, factor XIII, and von Willebrand factor. Due to infectious risk, the use of cryoprecipitate in Central Europe diminished over the last decades. However, after the introduction of various pathogen-reduction technologies for plasma, cryoprecipitate production in blood centers is a feasible alternative to pharmaceutical fibrinogen concentrate with a high safety profile. In our study, we evaluated the feasibility of the production of twenty-four cryoprecipitate units from pools of two units of apheresis plasma pathogen reduced using amotosalen and ultraviolet light A (UVA) (INTERCEPT^® Blood System). The aim was to assess the compliance of the pathogen-reduced cryoprecipitate with the European Directorate for the Quality of Medicines (EDQM) guidelines and the stability of coagulation factors after frozen (≤−25 °C) storage and five-day liquid storage at ambient temperature post-thawing. All pathogen-reduced cryoprecipitate units fulfilled the European requirements for fibrinogen, factor VIII and von Willebrand factor content post-preparation. After five days of liquid storage, content of these factors exceeded the minimum values in the European requirements and the content of other factors was sufficient. Our method of production of cryoprecipitate using pathogen-reduced apheresis plasma in a jumbo bag is feasible and efficient. Full article

(This article belongs to the Special Issue Pathogen Reduction of Blood Bank Components)

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Pathogen-reduced Cryoprecipitate (PR-Cryo). Coagulation factor stability after storage at <−25 °C and up to 5 days at ambient temperature (n = 24): (A) Fibrinogen, (B) Factor VIII, (C) vWF, (D) Factor XIII, (E) ADAMTS13, (F) Thromboelastography—Max. Amplitude after 30 min, and (G) Thrombin generation assay. (A–C) are the markers required by the EU regulatory body. (D–G) are markers not required by the EU regulatory body. “◊” represent mean values. “–” represent median values. “o” represents outliers. p-values were calculated from two-way mixed ANOVA model with frozen storage as between-product factor and room temperature (RT) storage as within-product factor. A p-value less than 0.05 indicates a significant effect of 24-month frozen storage or 5-day RT storage; significance is indicated by an asterisk (*). Full article ">Figure 1 Cont.
Pathogen-reduced Cryoprecipitate (PR-Cryo). Coagulation factor stability after storage at <−25 °C and up to 5 days at ambient temperature (n = 24): (A) Fibrinogen, (B) Factor VIII, (C) vWF, (D) Factor XIII, (E) ADAMTS13, (F) Thromboelastography—Max. Amplitude after 30 min, and (G) Thrombin generation assay. (A–C) are the markers required by the EU regulatory body. (D–G) are markers not required by the EU regulatory body. “◊” represent mean values. “–” represent median values. “o” represents outliers. p-values were calculated from two-way mixed ANOVA model with frozen storage as between-product factor and room temperature (RT) storage as within-product factor. A p-value less than 0.05 indicates a significant effect of 24-month frozen storage or 5-day RT storage; significance is indicated by an asterisk (*). Full article ">Figure 2
(A) Flow chart of plasma pathogen reduction. PR-plasma in the three containers is transferred into the jumbo ICPC bag after sterile connection. Afterwards, the ICPC bag and the three empty containers are shock-frozen together (not illustrated). (B) After being thawed and centrifuged the Cryo-poor supernatant is transferred into two of the attached plasma-storage containers; the PR-Cryo is transferred to the last plasma-storage container with subsequent freezing at <−25 °C. Full article ">

16 pages, 1002 KiB

Open AccessBrief Report

Improving Drug Sensitivity of HIV-1 Protease Inhibitors by Restriction of Cellular Efflux System in a Fission Yeast Model

by Jiantao Zhang, Qi Li, Shigehiro A. Kawashima, Mohamed Nasr, Fengtian Xue and Richard Y. Zhao

Pathogens 2022, 11(7), 804; https://doi.org/10.3390/pathogens11070804 - 16 Jul 2022

Cited by 2 | Viewed by 2020

Abstract

Fission yeast can be used as a cell-based system for high-throughput drug screening. However, higher drug concentrations are often needed to achieve the same effect as in mammalian cells. Our goal here was to improve drug sensitivity so reduced drugs could be used. [...] Read more.

Fission yeast can be used as a cell-based system for high-throughput drug screening. However, higher drug concentrations are often needed to achieve the same effect as in mammalian cells. Our goal here was to improve drug sensitivity so reduced drugs could be used. Three different methods affecting drug uptakes were tested using an FDA-approved HIV-1 protease inhibitor (PI) drug Darunavir (DRV). First, we tested whether spheroplasts without cell walls increase the drug sensitivity. Second, we examined whether electroporation could be used. Although small improvements were observed, neither of these two methods showed significant increase in the EC₅₀ values of DRV compared with the traditional method. In contrast, when DRV was tested in a mutant strain PR836 that lacks key proteins regulating cellular efflux, a significant increase in the EC₅₀ was observed. A comparison of nine FDA-approved HIV-1 PI drugs between the wild-type RE294 strain and the mutant PR836 strain showed marked enhancement of the drug sensitivities ranging from an increase of 0.56 log to 2.48 logs. Therefore, restricting cellular efflux through the adaption of the described fission yeast mutant strain enhances the drug sensitivity, reduces the amount of drug used, and increases the chance of success in future drug discovery. Full article

(This article belongs to the Collection Novel Strategies on Antiviral Drug Discovery Against Human Diseases)

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12 pages, 2208 KiB

Open AccessArticle

Establishing a Herpesvirus Quiescent Infection in Differentiated Human Dorsal Root Ganglion Neuronal Cell Line Mediated by Micro-RNA Overexpression

by Yu-Chih Chen, Hedong Li, Miguel Martin-Caraballo and Shaochung Victor Hsia

Pathogens 2022, 11(7), 803; https://doi.org/10.3390/pathogens11070803 - 16 Jul 2022

Cited by 2 | Viewed by 2490

Abstract

HSV-1 is a neurotropic pathogen associated with severe encephalitis, excruciating orofacial sensation, and other chronic neuropathic complications. After the acute infection, the virus may establish a lifelong latency in the neurons of trigeminal ganglia (TG) and other sensory and autonomic ganglia, including the [...] Read more.

HSV-1 is a neurotropic pathogen associated with severe encephalitis, excruciating orofacial sensation, and other chronic neuropathic complications. After the acute infection, the virus may establish a lifelong latency in the neurons of trigeminal ganglia (TG) and other sensory and autonomic ganglia, including the dorsal root ganglia (DRG), etc. The reactivation occurred periodically by a variety of physical or emotional stressors. We have been developing a human DRG neuronal cell-culture model HD10.6, which mimics the mature neurons for latency and reactivation with robust neuronal physiology. We found that miR124 overexpression without acyclovir (ACV) could maintain the virus in a quiescent infection, with the accumulation of latency-associate transcript (LAT). The immediate-early (IE) gene ICP0, on the other hand, was very low and the latent viruses could be reactivated by trichostatin A (TSA) treatment. Together, these observations suggested a putative role of microRNA in promoting HSV-1 latency in human neurons. Full article

(This article belongs to the Special Issue Host–Virus Interactions in the Nervous System)

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Figure 1
miR-124 is overexpressed in HEK-293 and HD10.6 cells following L124 transduction, performed in triplicate. The mature miR-124 was introduced into HEK-293 cells by L124 infection. The miR-124 expression level was validated by qRT-PCR. miR-124 expression appeared to increase by approximately 10-fold (A). HD10.6 cells were first transduced with L124 followed by HSV-1 infections at MOI of 1. The miR-124 expression was analyzed by the same method described in A. Quantitative analyses of miR-124 expression indicate a 3-fold and 4.95-fold upsurge with or without HSV-1 infection, respectively (B). All qRT-PCR experiments were normalized to miR-26a in a quadruple fashion for statistical analyses. The asterisk (*) designates significant differences compared to vector or no L124 control. Note that both A and B were normalized to miR-26a. The viral titer of the <a href="#pathogens-11-00803-f001" class="html-fig">Figure 1</a>B supernatant is 3.6 × 103 PFU/mL for the HD10.6 + HSV-1 group, while the others are less than 100 PFU/mL (determined by standard plaque assay; data not shown). Full article ">Figure 2
Differentiated HD10.6 cells with miR124 overexpression can be co-infected by HSV-1. The L124-transduced HD10.6 cells (HD-L-Stable) were subjected to KVP-mRFP HSV-1 infection at the MOI of 1 followed by fluorescent microscopy. The green and red fluorescence represented the miR124 expression and HSV-1 infection, respectively. The merged image demonstrated the presence of miR-124 in the HSV-1 infected cells. The size of the scale bar is 1 mm. Full article ">Figure 3
Electrophysiology of infected HD10.6 cells following miR-124 overexpression. Electrophysiological studies indicated that overexpression of miR-124 evokes a 47% decrease in cell capacitance, suggesting a decrease in the size of HD10.6 cells. The y-axis denotes Capacitance (pF) (A). Analysis of the sodium current densities demonstrated no significant difference between control and HD-L-Stable cells. HSV-1 acute infection decreases the functional expression of sodium channels. The y-axis denotes current density (pA/pF) (B). NS: denotes “Not significant”; * denotes statistically significant differences. Full article ">Figure 4
A dormant state of HSV-1 infection was achieved with miR-124 overexpression. Differentiated HD10.6 cells, with or without L124 transduction, were subjected to HSV-1 infection, in the absence of ACV, at the MOI of 1 followed by plaque assays using the media supernatant collected daily for three days. It is visible that HSV-1 replicated actively in HD10.6 cells without miR-124. The HD-L-Stable cells, however, released fewer viral particles (A). Other infections were performed with the MOI of 0.15 followed by the same analyses. It appeared that the infectious viruses were not released until 6 dpi in regular HD10.6 cells, and no release of infectious virus was detected in HD-L-Stable cells. Note that the assays were measured at 2, 4, and 6 dpi due to much lower titers used for infection (B). All plaque assay experiments were performed in triplicate for statistical analyses. Full article ">Figure 5
HSV-1 was not released but present within miR124 overexpressing HD10.6 cells. HD-L-Stable cells in a latency-like state were subjected to reactivation by the histone deacetylase inhibitor TSA. No release of infectious viruses was detected in the media supernatant of the HD-L-Stable cells, even after TSA treatment. In the regular HD10.6 cells, infectious viruses from the media supernatant were spotted in two out of three wells and the number increased after TSA treatment (A). The cell lysate was subjected to plaque assays, and the infectious viral particles were observed in both cases, but HD-L-Stable cells exhibited approximately a 3.8-fold reduction in plaque formation compared to the original HD10.6 cells (compare orange to yellow spots). The TSA was sufficient to boost the viral replication in both cases. All plaque assay experiments were performed in sextuplicate followed by dot plot analyses and the statistical analyses by ANOVA indicated the differences were significant (denoted by *) (B). Full article ">Figure 6
Viral transcription increased after TSA-induced reactivation from the dormant infection. The effects of TSA to reactivate the viral gene expression from HD-L-Stable cells were measured by qRT-PCR. The transcripts of TK were quite weak but increased approximately 7.2-fold after TSA treatment (A). The ICP0 transcription exhibited a similar pattern with 7.8-fold increases upon TSA reactivation (B). The miR-124 expression maintained a similar level after the treatment (C). Note that, in (A,B), the gene induction by TSA varied probably due to the different copy numbers of viruses maintained during latency. Full article ">Figure 7
Viral transcription increased after TSA-induced reactivation from the dormant infection. The HSV-1 induced gene LAT accumulates in HD10.6 cells overexpressing miR-124. The accumulation of HSV-1 LAT was analyzed by qRT-PCR. No difference was observed at 3 dpi but a significant increase appeared in HD-L-Stable cells at 6 dpi (orange line). The LAT accumulation in regular HD10.6 cells is displayed as a blue line (A). The accumulation of LAT showed no difference after TSA treatment (B). * denotes statistically significant differences. Full article ">Figure 8
Structure of miR-124 and the putative binding to HSV-1 UL36. The stem-loop structure of miR-124 was depicted. The functional domain was labeled in red (A). The putative match by computational analyses was described and the results exhibited two hits with 100% match. The miR-124 was first matched from 48–59 to UL36 cDNA 1545–1556 (B). The second match of miR-124 was from 52–61 to UL36 cDNA 4232–4241 (C). The HSV-1 sequence is based on the strain McKrae (GenBank#: MN136524.1). Full article ">

13 pages, 900 KiB

Open AccessArticle

Anthroponotic-Based Transfer of Staphylococcus to Dog: A Case Study

by Massimiliano Orsini, Sara Petrin, Michela Corrò, Giulia Baggio, Elena Spagnolo and Carmen Losasso

Pathogens 2022, 11(7), 802; https://doi.org/10.3390/pathogens11070802 - 15 Jul 2022

Cited by 4 | Viewed by 2039

Abstract

Although usually harmless, Staphylococcus spp. can cause nosocomial and community-onset skin and soft tissue infections in both humans and animals; thus, it is considered a significant burden for healthcare systems worldwide. Companion animals have been identified as potential reservoirs of pathogenic Staphylococcus with [...] Read more.

Although usually harmless, Staphylococcus spp. can cause nosocomial and community-onset skin and soft tissue infections in both humans and animals; thus, it is considered a significant burden for healthcare systems worldwide. Companion animals have been identified as potential reservoirs of pathogenic Staphylococcus with specific reference to Methicillin Resistant Staphylococcus aureus (MRSA). In this study, we investigated the circulation and the genetic relationships of a collection of Staphylococcus spp. isolates in a family composed of four adults (a mother, father, grandmother, and grandfather), one child, and a dog, which were sampled over three years. The routes of transmission among humans and between humans and the dog werelyzed. The results displayed the circulation of many Staphylococcus lineages, belonging to different species and sequence types (ST) and being related to both human and pet origins. However, among the observed host-switch events, one of them clearly underpinnthroponotic route from a human to a dog. This suggests that companion animals can potentially have a role as a carrier of Staphylococcus, thus posing a serious concern about MRSA spreading within human and animal microbial communities. Full article

(This article belongs to the Special Issue Exploring the Epidemiology, Pathogenicity, and Therapeutic Options of Staphylococcus spp.)

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16 pages, 2866 KiB

Open AccessEditor’s ChoiceArticle

Complete Genomes of Theileria orientalis Chitose and Buffeli Genotypes Reveal within Species Translocations and Differences in ABC Transporter Content

by Jerald Yam, Daniel R. Bogema, Melinda L. Micallef, Steven P. Djordjevic and Cheryl Jenkins

Pathogens 2022, 11(7), 801; https://doi.org/10.3390/pathogens11070801 - 15 Jul 2022

Cited by 3 | Viewed by 2784

Abstract

Theileria orientalis causes losses to cattle producers in Eastern Asia, Oceania and, more recently, North America. One pathogenic genotype (Ikeda) has been sequenced to the chromosomal level, while only draft genomes exist for globally distributed Chitose and Buffeli genotypes. To provide an accurate [...] Read more.

Theileria orientalis causes losses to cattle producers in Eastern Asia, Oceania and, more recently, North America. One pathogenic genotype (Ikeda) has been sequenced to the chromosomal level, while only draft genomes exist for globally distributed Chitose and Buffeli genotypes. To provide an accurate comparative gene-level analysis and help further understand their pathogenicity, we sequenced isolates of the Chitose and Buffeli genotypes of T. orientalis using long-read sequencing technology. A combination of several long-read assembly methods and short reads produced chromosomal-level assemblies for both Fish Creek (Chitose) and Goon Nure (Buffeli) isolates, including the first complete and circular apicoplast genomes generated for T. orientalis. Comparison with the Shintoku (Ikeda) reference sequence showed both large and small translocations in T. orientalis Buffeli, between chromosomes 2 and 3 and chromosomes 1 and 4, respectively. Ortholog clustering showed expansion of ABC transporter genes in Chitose and Buffeli. However, differences in several genes of unknown function, including DUF529/FAINT-domain-containing proteins, were also identified and these genes were more prevalent in Ikeda and Chitose genotypes. Phylogenetics and similarity measures were consistent with previous short-read genomic analysis. The generation of chromosomal sequences for these highly prevalent T. orientalis genotypes will also support future studies of population genetics and mixed genotype infections. Full article

(This article belongs to the Special Issue Bovine Theileriosis Caused by the Theileria orientalis Group)

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Figure 1
Apicoplast genomes of T. orientalis Fish Creek and Goon Nure isolates. Outer ring (black) represents DNA sequence. Middle ring shows annotated genes including ribosomal RNA subunits (red), transfer RNA (purple) and protein coding sequences (green). Inner ring shows %GC difference from average with a 100 bp sliding window. Full article ">Figure 2
Synteny dot plots of the T. orientalis Shintoku (Ikeda) reference and strains Fish Creek (Chitose) and Goon Nure (Buffeli). Red circles indicate rearrangement in strain Goon Nure between chromosomes 2 and 3 and translocation between chromosomes 1 and 4. Full article ">Figure 2 Cont.
Synteny dot plots of the T. orientalis Shintoku (Ikeda) reference and strains Fish Creek (Chitose) and Goon Nure (Buffeli). Red circles indicate rearrangement in strain Goon Nure between chromosomes 2 and 3 and translocation between chromosomes 1 and 4. Full article ">Figure 3
Venn diagram showing number of genes found in each isolate combination. Full article ">Figure 4
COG analysis of all predicted genes (top); genes identified as unique to each isolate combination (middle). Genes without COG assignment are not shown but consist of 31–38% of the total gene content of each isolate. COG categories (x-axis) are summarised by their letter categories (bottom). Full article ">Figure 5
Maximum likelihood tree of Piroplasmida whole-genome protein sequences inferred with concordance factors with IQ-TREE 2 using 1417 concatenated protein sequences from single-copy genes. P. vivax str. Salvador I and P. falciparum str. 3D7 were used as outgroups. Each branch label on the tree shows the bootstrap, gene concordance factor (gCF) and site concordance factor (sCF), respectively (bootstrap/gCF/sCF). Full article ">

42 pages, 2769 KiB

Open AccessReview

RNA Viruses, Pregnancy and Vaccination: Emerging Lessons from COVID-19 and Ebola Virus Disease

by Chandrasekharan Rajalekshmi Dhanya, Aswathy Shailaja, Aarcha Shanmugha Mary, Sumodan Padikkala Kandiyil, Ambili Savithri, Vishnu Sasidharan Lathakumari, Jayakrishnan Therthala Veettil, Jiji Joseph Vandanamthadathil and Maya Madhavan

Pathogens 2022, 11(7), 800; https://doi.org/10.3390/pathogens11070800 - 15 Jul 2022

Cited by 3 | Viewed by 4877

Abstract

Pathogenic viruses with an RNA genome represent a challenge for global human health since they have the tremendous potential to develop into devastating pandemics/epidemics. The management of the recent COVID-19 pandemic was possible to a certain extent only because of the strong foundations [...] Read more.

Pathogenic viruses with an RNA genome represent a challenge for global human health since they have the tremendous potential to develop into devastating pandemics/epidemics. The management of the recent COVID-19 pandemic was possible to a certain extent only because of the strong foundations laid by the research on previous viral outbreaks, especially Ebola Virus Disease (EVD). A clear understanding of the mechanisms of the host immune response generated upon viral infections is a prime requisite for the development of new therapeutic strategies. Hence, we present here a comparative study of alterations in immune response upon SARS-CoV-2 and Ebola virus infections that illustrate many common features. Vaccination and pregnancy are two important aspects that need to be studied from an immunological perspective. So, we summarize the outcomes and immune responses in vaccinated and pregnant individuals in the context of COVID-19 and EVD. Considering the significance of immunomodulatory approaches in combating both these diseases, we have also presented the state of the art of such therapeutics and prophylactics. Currently, several vaccines against these viruses have been approved or are under clinical trials in various parts of the world. Therefore, we also recapitulate the latest developments in these which would inspire researchers to look for possibilities of developing vaccines against many other RNA viruses. We hope that the similar aspects in COVID-19 and EVD open up new avenues for the development of pan-viral therapies. Full article

(This article belongs to the Special Issue Host Immune Responses to RNA Viruses)

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Innate immune response alterations common for SARS-CoV-2 and EBOV infections. Full article ">Figure 2
Comparative antibody response curve upon SARS-CoV-2 and EBOV infection. Full article ">Figure 3
Mechanisms of Lymphopenia in SARS-CoV-2 (A) and EBOV (B) infections. Lymphopenia, a common feature of COVID-19 and EVD, is characterized by different routes of T-cell death such as apoptosis, necrosis, necroptosis and autophagy. (A,B) demonstrate the known mechanisms involved in each of these pathways in COVID-19 and EVD, respectively. It is noteworthy that many of these alterations are common for both the diseases, the fact that can be exploited for the design of novel pan-viral therapies. Full article ">Figure 4
Molecular alterations associated with lymphopenia in COVID-19 patients. Proapoptotic BCl2 proteins, CXCL10, CCL2 and IL6 are upregulated. Antiviral antibodies IgG and IgM, infection of bone marrow progenitors and the upregulated IL6 suppresses lymphopoiesis. IL6 also suppresses the thymus. All these are reported to be associated with lymphopenia in COVID-19 patients. Full article ">Figure 5
Factors associated with T-cell exhaustion in COVID-19 and EVD. Proliferation of T-cells, release of cytokines, cytotoxic and self-renewal capabilities of T-cells and glycolysis are commonly found to be reduced in COVID-19 and EVD. Additionally, T-cells express exhaustion markers, of which PD-1 and CTLA4 are common. Alteration of transcription of genes related to TCR and cytokine signaling pathways and dysregulation of mitochondrial energetics are also a common feature of T-cell exhaustion in COVID-19 and EVD. Full article ">Figure 6
Alterations in immune response in pregnant women during viral infections. Full article ">

13 pages, 4211 KiB

Open AccessArticle

Productive Replication of HIV-1 but Not SIVmac in Small Ruminant Cells

by Hibet Errahmane Chergui, Takfarinas Idres, Chloé Chaudesaigues, Diana Noueihed, Jean Gagnon and Yahia Chebloune

Pathogens 2022, 11(7), 799; https://doi.org/10.3390/pathogens11070799 - 15 Jul 2022

Viewed by 2667

Abstract

Animal lentiviruses (LVs) have been proven to have the capacity to cross the species barrier, to adapt in the new hosts, and to increase their pathogenesis, therefore leading to the emergence of threatening diseases. However, their potential for widespread diffusion is limited by [...] Read more.

Animal lentiviruses (LVs) have been proven to have the capacity to cross the species barrier, to adapt in the new hosts, and to increase their pathogenesis, therefore leading to the emergence of threatening diseases. However, their potential for widespread diffusion is limited by restrictive cellular factors that block viral replication in the cells of many species. In previous studies, we demonstrated that the restriction of CAEV infection of sheep choroid plexus cells was due to aberrant post-translation cleavage of the CAEV Env gp170 precursor. Later, we showed that the lack of specific receptor(s) for caprine encephalitis arthritis virus (CAEV) on the surface of human cells was the only barrier to their infection. Here, we examined whether small ruminant (SR) cells can support the replication of primate LVs. Three sheep and goat cell lines were inoculated with cell-free HIV-1 and SIVmac viral stocks or transfected with infectious molecular clone DNAs of these viruses. The two recombinant lentiviral clones contained the green fluorescent protein (GFP) reporter sequence. Infection was detected by GFP expression in target cells, and the infectious virus produced and released in the culture medium of treated cells was detected using the indicator TZM-bl cell line. Pseudotyped HIV-GFP and SIV-GFP with vesicular stomatitis virus G glycoprotein (VSV-G) allowed the cell receptors to be overcome for virus entry to further evaluate the viral replication/restriction in SR cells. As expected, neither HIV nor SIV viruses infected any of the SR cells. In contrast, the transfection of plasmid DNAs of the infectious molecular clones of both viruses in SR cells produced high titers of infectious viruses for human indicators, but not SR cell lines. Surprisingly, SR cells inoculated with HIV-GFP/VSV-G, but not SIV-GFP/VSV-G, expressed the GFP and produced a virus that efficiently infected the human indictor, but not the SR cells. Collectively, these data provide a demonstration of the lack of replication of the SIVmac genome in SR cells, while, in contrast, there was no restriction on the replication of the IV-1 genome in these cells. However, because of the lack of functional receptors to SIVmac and HIV-1 at the surface of SR cells, there is specific lentiviral entry. Full article

(This article belongs to the Special Issue Animal Retrovirus)

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Organization of pHIV-GFP and pSIV-GFP plasmid DNAs genomes. Full article ">Figure 2
Detection of GFP expression in pHIV-GFP and pSIV-GFP transfected cells. (A) HEK, CRFK and TZM-bl cell lines. (B) TIGEF, TYGSM and RMI cell lines. Cell monolayers were transfected as described in Materials and Methods. At 24 h post-transfection, the cell monolayers were observed under a fluorescence microscope to assess the expression of GFP. (c.1–c.6) Cells transfected with pHIV-GFP. (d.1–d.6) Cells transfected with pSIV-GFP. (a.1–a.6) Non-transfected cells were used as a negative control. (c.3) TZM-bl cells transfected with pHIV-GFP and (d.3) pSIV-GFP were used as a positive control along with cells transfected with GFP plasmid (b.1–b.6). The images were acquired as a merge of the green channel and the bright field. Acquisitions were performed with 488 nm excitation and the emission was collected at 500–600 nm. Full article ">Figure 3
Detection of SIV-GFP and HIV-GFP infection by fluorescence microscopy. (A) HEK, CRFK and TZM-bl human and feline cell lines. (B) TIGEF, TYGSM and RMI SR cell lines. Monolayers of each of the cell lines were inoculated with HIV-1 and SIVmac viral stocks expressing GFP as indicated in Materials and Methods. At 120 h post-infection, the monolayers were observed under a fluorescence microscope to assess the expression of GFP. (b.1–b.3) Cells inoculated with HIV-GFP. (c.1–c.3) Cells inoculated with SIV-GFP. (a.1–a.3) Non-inoculated cell lines were used as a negative control. (b.3) TZM-bl cells inoculated with SIV-GFP and (c.3) HIV-GFP were used as positive controls. The images are a merge of the green channel and the bright field. They were acquired with 488 nm excitation and the emission was collected at 500–600 nm. Full article ">Figure 4
Detection of HIV-GFP/VSV-G and SIV-GFP/VSV-G infection by fluorescence microscopy. (A) HEK, CRFK and TZM-bl cell lines. (B) TIGEF, TYGSM and RMI cell lines. The cell lines were inoculated with SIV-GFP and HIV-GFP pseudotyped with VSV-G. At 120 h post-inoculation, the cell lines were observed under a fluorescence microscope to assess GFP expression in the monolayers. (b.1–b.6) Cells inoculated with HIV-GFP/VSV-G. (c.1–c.6) Cells inoculated with SIV-GFP/VSV-G. (a.1–a.6) Non-transfected cell lines were used as a negative control. TZM-bl cells inoculated with (b.3) HIV-GFP/VSV-G and (c.3) SIV-GFP/VSV-G were used as a positive control, respectively. The images are a merge of the green channel and the bright field. They were acquired with 488 nm excitation and the emission was collected at 500–600 nm. Full article ">Figure 4 Cont.
Detection of HIV-GFP/VSV-G and SIV-GFP/VSV-G infection by fluorescence microscopy. (A) HEK, CRFK and TZM-bl cell lines. (B) TIGEF, TYGSM and RMI cell lines. The cell lines were inoculated with SIV-GFP and HIV-GFP pseudotyped with VSV-G. At 120 h post-inoculation, the cell lines were observed under a fluorescence microscope to assess GFP expression in the monolayers. (b.1–b.6) Cells inoculated with HIV-GFP/VSV-G. (c.1–c.6) Cells inoculated with SIV-GFP/VSV-G. (a.1–a.6) Non-transfected cell lines were used as a negative control. TZM-bl cells inoculated with (b.3) HIV-GFP/VSV-G and (c.3) SIV-GFP/VSV-G were used as a positive control, respectively. The images are a merge of the green channel and the bright field. They were acquired with 488 nm excitation and the emission was collected at 500–600 nm. Full article ">Figure 5
Flow cytometry analyses of cells inoculated with VSV-G pseudotyped HIV-GFP and SIV-GFP. Cells were acquired in a FACSCantoII and displayed according to FSC/GFP characteristics. Cells were analyzed using FlowJo software. Non-inoculated cells were used as negative controls. Full article ">

15 pages, 2997 KiB

Open AccessArticle

Ginger Is a Potential Therapeutic for Chronic Toxoplasmosis

by Asmaa M. El-kady, Wafa Abdullah I. Al-Megrin, Iman A. M. Abdel-Rahman, Eman Sayed, Eman Abdullah Alshehri, Majed H. Wakid, Fadi M. Baakdah, Khalil Mohamed, Hayam Elshazly, Hussah M. Alobaid, Safa H. Qahl, Hatem A. Elshabrawy and Salwa S. Younis

Pathogens 2022, 11(7), 798; https://doi.org/10.3390/pathogens11070798 - 15 Jul 2022

Cited by 6 | Viewed by 6089

Abstract

Background:Toxoplasma gondii (T. gondii) is an opportunistic parasite that causes serious diseases in humans, particularly immunocompromised individuals and pregnant women. To date, there are limited numbers of therapeutics for chronic toxoplasmosis which necessitate the discovery of effective and safe therapeutics. [...] Read more.

Background:Toxoplasma gondii (T. gondii) is an opportunistic parasite that causes serious diseases in humans, particularly immunocompromised individuals and pregnant women. To date, there are limited numbers of therapeutics for chronic toxoplasmosis which necessitate the discovery of effective and safe therapeutics. In the present study, we aimed to evaluate the antitoxoplasmosis potential of ginger extract in mice with experimentally induced chronic toxoplasmosis. Results: Treatment with ginger extract significantly reduced cysts count in the brains of T. gondii-infected mice with a marked alleviation of edema and inflammation, and a reversal of neuronal injury. Moreover, ginger extract treatment reduced inflammation in liver and lungs and protected hepatocytes from infection-induced degeneration. Consistently, apoptosis was significantly mitigated in the brains of ginger extract-treated mice compared to infected untreated animals or spiramycin-treated animals. Methods: Four groups of Swiss albino mice (10 mice each) were used. The first group was not infected, whereas 3 groups were infected with Me49 T. gondii strains. One infected group remained untreated (infected untreated), whereas the other two infected groups were treated with either ginger extract (250 mg/kg) or spiramycin (positive control; 100 mg/kg), respectively. The therapeutic potential of ginger extract was evaluated by calculation of the parasite burden in infected animals, and examination of the infected tissues for reduced pathologic changes. Conclusions: Our results showed for the first time that ginger extract exhibited marked therapeutic effects in mice with chronic T. gondii infection which indicates that it can be used as a safe and effective treatment for chronic toxoplasmosis. Full article

(This article belongs to the Special Issue Optimizing Treatment for Parasitic Infections)

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8 pages, 1875 KiB

Open AccessCommunication

First Report of a Complete Genome Sequence of a Variant African Swine Fever Virus in the Mekong Delta, Vietnam

by Nguyen Duc Hien, Lam Thanh Nguyen, Le Trung Hoang, Nguyen Ngoc Bich, To My Quyen, Norikazu Isoda and Yoshihiro Sakoda

Pathogens 2022, 11(7), 797; https://doi.org/10.3390/pathogens11070797 - 15 Jul 2022

Cited by 7 | Viewed by 2960

Abstract

The objective of this study is to report the complete-genome sequence of a field African swine fever (ASF) virus (ASFV), namely ASF/VN/CanTho-OM/2021, which caused a fatal outbreak in domestic pigs in the Mekong Delta. Complete-genome sequencing detected an 18 bp nucleotide deletion in [...] Read more.

The objective of this study is to report the complete-genome sequence of a field African swine fever (ASF) virus (ASFV), namely ASF/VN/CanTho-OM/2021, which caused a fatal outbreak in domestic pigs in the Mekong Delta. Complete-genome sequencing detected an 18 bp nucleotide deletion in the EP402R gene (encoding for serotype-specific proteins CD2v) of ASF/VN/CanTho-OM/2021, which was determined to belong to genotype 2 and serotype 8. This mutation pattern was confirmed as unique in GenBank; thus, ASF/VN/CanTho-OM/2021 can be considered a novel variant, with a potential change of sero-characteristics within genotype 2. An additional unique mutation of 78 bp nucleotide insertion was also observed in the B475L gene. Additionally, four copies of tandem repeat sequences were found in the intergenic region (IGR) located between I73R and I329L, previously assigned as the IGR III variant. This study is the first to report the complete genome of ASFV in the Mekong Delta, and it highlights the necessity of strengthening molecular surveillance to provide further knowledge on the evolution and incursion of ASFV in the Mekong Delta and Vietnam. Full article

(This article belongs to the Collection Emerging and Re-emerging Pathogens)

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14 pages, 1503 KiB

Open AccessReview

Gut Microbes and Neuropathology: Is There a Causal Nexus?

by Katherine Dinan and Timothy G. Dinan

Pathogens 2022, 11(7), 796; https://doi.org/10.3390/pathogens11070796 - 14 Jul 2022

Cited by 9 | Viewed by 3623

Abstract

The gut microbiota is a virtual organ which produces a myriad of molecules that the brain and other organs require. Humans and microbes are in a symbiotic relationship, we feed the microbes, and in turn, they provide us with essential molecules. Bacteroidetes and [...] Read more.

The gut microbiota is a virtual organ which produces a myriad of molecules that the brain and other organs require. Humans and microbes are in a symbiotic relationship, we feed the microbes, and in turn, they provide us with essential molecules. Bacteroidetes and Firmicutes phyla account for around 80% of the total human gut microbiota, and approximately 1000 species of bacteria have been identified in the human gut. In adults, the main factors influencing microbiota structure are diet, exercise, stress, disease and medications. In this narrative review, we explore the involvement of the gut microbiota in Parkinson’s disease, Alzheimer’s disease, multiple sclerosis and autism, as these are such high-prevalence disorders. We focus on preclinical studies that increase the understanding of disease pathophysiology. We examine the potential for targeting the gut microbiota in the development of novel therapies and the limitations of the currently published clinical studies. We conclude that while the field shows enormous promise, further large-scale studies are required if a causal link between these disorders and gut microbes is to be definitively established. Full article

(This article belongs to the Collection Current Status of Research on Gut Metabolites and Microbiota)

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11 pages, 1199 KiB

Open AccessArticle

Pathogenicity and Metabolites of Purpureocillium lavendulum YMF1.00683 against Meloidogyne incognita

by Zheng-Xue Bao, Rui Liu, Chun-Qiang Li, Xue-Rong Pan and Pei-Ji Zhao

Pathogens 2022, 11(7), 795; https://doi.org/10.3390/pathogens11070795 - 14 Jul 2022

Cited by 3 | Viewed by 2094

Abstract

Purpureocillium lavendulum is a biological control agent with several registered products that can parasitize the eggs and larvae of various pathogenic nematodes. In this study, the pathogenicity and secondary metabolites of the fungus P. lavendulum YMF1.00683 were investigated. The strain YMF1.00683 had infection [...] Read more.

Purpureocillium lavendulum is a biological control agent with several registered products that can parasitize the eggs and larvae of various pathogenic nematodes. In this study, the pathogenicity and secondary metabolites of the fungus P. lavendulum YMF1.00683 were investigated. The strain YMF1.00683 had infection efficiency against the plant root-knot nematode Meloidogyne incognita. The strain’s process of infecting nematodes was observed under a microscope. Moreover, seven metabolites, including a new sterol (1), were isolated and identified from cultures of YMF1.0068 in Sabouraud’s dextrose agar. A bioassay showed that 5-methoxymethyl-1H-pyrrole-2-carboxaldehyde (7) is toxic to M. incognita and affects the egg hatching. It caused 98.23% mortality in M. incognita and could inhibit 80.78% of the hatching eggs at 400 μg/mL over a period of 96 h. Furthermore, 5-methoxymethyl-1H-pyrrole-2-carboxaldehyde (7) showed a strong avoidance effect at 40 ppm, and its chemotactic index value was −0.37. The results indicate that P. lavendulum could produce active metabolites against M. incognita. Full article

(This article belongs to the Special Issue Microbe-Nematode Interactions)

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15 pages, 2567 KiB

Open AccessArticle

Characterization of Bordetella pertussis Strains Isolated from India

by Shweta Alai, Manish Gautam, Sonali Palkar, Jitendra Oswal, Sunil Gairola and Dhiraj P. Dhotre

Pathogens 2022, 11(7), 794; https://doi.org/10.3390/pathogens11070794 - 14 Jul 2022

Cited by 2 | Viewed by 2879

Abstract

Despite high level vaccination and the availability of two different types of vaccines, whole cell (wP) and acellular vaccines (aP), the resurgence of pertussis has been reported in many countries. Antigenic variation within circulating and vaccine strains is the most documented reason reported [...] Read more.

Despite high level vaccination and the availability of two different types of vaccines, whole cell (wP) and acellular vaccines (aP), the resurgence of pertussis has been reported in many countries. Antigenic variation within circulating and vaccine strains is the most documented reason reported for the resurgence of pertussis. Research on genetic divergence among circulating and vaccine strains has largely been reported in countries using aP vaccines. There are inadequate data available for antigenic variation in B. pertussis from wP-using countries. India has used wP for more than 40 years in their primary immunization program. The present study reports five clinical isolates of B. pertussis from samples of pediatric patients with pertussis symptoms observed in India. Genotypic and phenotypic characterization of clinical isolates were performed by serotyping, genotyping, whole genome analyses and comparative genomics. All clinical isolates showed serotype 1, 2 and 3 based on the presence of fimbriae 2 and 3. Genotyping showed genetic similarities in allele types for five aP genes within vaccine strains and clinical isolates reported from India. The presence of the ptxP3 genotype was observed in two out of five clinical isolates. Whole-genome sequencing was performed for clinical isolates using the hybrid strategy of combining Illumina (short reads) and oxford nanopore (long reads) sequencing strategies. Clinical isolates (n = 5) and vaccine strains (n = 7) genomes of B. pertussis from India were compared with 744 B. pertussis closed genomes available in the public databases. The phylogenomic comparison of B. pertussis genomes reported from India will be advantageous in better understanding pertussis resurgence reported globally with respect to pathogen adaptation. Full article

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Agarose gel electrophoresis (2%) of PCR amplified products using virulence and species-specific PCR primer sets. Lanes 2–17 are examined for B. pertussis isolates. Lanes 2–17 from Isolate S1 for prn, ompQ, cyaA, TcfA, PtxA, BapC, Fim3, Fim2, vag8, BrkA, PtxP, Pgm, FHA, IS481, IS1002, IS1001, respectively. Lane 1: 0.1–10 kb DNA size marker, Lane 20: 100bp DNA size marker. Lane 18: Negative control. Full article ">Figure 2
Whole genome alignment of B. pertussis genomes from India (A) Genome alignment of vaccine and clinical isolates. Homologous blocks are represented by the same colour and connecting lines show rearrangements within homologous blocks. (B) Inversion were analyzed using NC_002929 (Tohama I) as reference with clinical isolate S5. Full article ">Figure 3
Global positioning of Indian B. pertussis genomes (A) Cut out section of phylogeny highlighting B. pertussis strains reported from India (B) Mash-based tree generated from whole genomes of all B. pertussis genomes using UPMGA highlighting sequenced types of isolates and vaccine strains from India S1, S2, S3, S4, S5. Green color indicate vaccine strains, red color indicate clinical isolates reported from India, Blue color indicate reference strain TohamaI. Full article ">

12 pages, 572 KiB

Open AccessArticle

Evolution of the Clinical Profile and Outcomes of Unvaccinated Patients Affected by Critical COVID-19 Pneumonia from the Pre-Vaccination to the Post-Vaccination Waves in Italy

by Cecilia Calabrese, Anna Annunziata, Domenica Francesca Mariniello, Antonietta Coppola, Angela Irene Mirizzi, Francesca Simioli, Corrado Pelaia, Lidia Atripaldi, Gaia Pugliese, Salvatore Guarino and Giuseppe Fiorentino

Pathogens 2022, 11(7), 793; https://doi.org/10.3390/pathogens11070793 - 14 Jul 2022

Cited by 2 | Viewed by 2041

Abstract

The vaccination campaign and the new SARS-CoV-2 variants may have changed the clinical profile and outcomes of patients admitted to sub-intensive unit care. We conducted a retrospective study aimed to compare the clinical and radiological features of unvaccinated critical COVID-19 patients hospitalized during [...] Read more.

The vaccination campaign and the new SARS-CoV-2 variants may have changed the clinical profile and outcomes of patients admitted to sub-intensive unit care. We conducted a retrospective study aimed to compare the clinical and radiological features of unvaccinated critical COVID-19 patients hospitalized during the last pandemic wave (December 2021–February 2022, No-Vax group) and before starting the vaccination campaign (March–December 2020, Pre-Vax group). The No-Vax group was also compared with vaccinated patients of the same pandemic wave (Vax group). With respect to the Pre-Vax group, the No-Vax group contained a higher percentage of smokers (p = 0.0007) and a lower prevalence of males (p = 0.0003). At admission, the No-Vax patients showed both a higher CT score of pneumonia and a worse severe respiratory failure (p < 0.0001). In the No-Vax group, a higher percentage of deaths occurred, though this was not significant. In comparison with the No-Vax group, the Vax patients were older (p = 0.0097), with a higher Charlson comorbidity index (p < 0.0001) and a significantly lower HRCT score (p = 0.0015). The percentage of deaths was not different between the two groups. The No-Vax patients showed a more severe disease in comparison with the Pre-Vax patients, and were younger and had fewer comorbidities than the Vax patients. Full article

(This article belongs to the Special Issue Infections and Over-Infections in COVID-19 Era: Diagnostics and Treatments)

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17 pages, 3021 KiB

Open AccessReview

Gurltia paralysans: A Neglected Angio-Neurotropic Parasite of Domestic Cats (Felis catus) and Free-Ranging Wild Felids (Leopardus spp.) in South America

by Lisbeth Rojas-Barón, Anja Taubert, Carlos Hermosilla, Marcelo Gómez, Manuel Moroni and Pamela Muñoz

Pathogens 2022, 11(7), 792; https://doi.org/10.3390/pathogens11070792 - 13 Jul 2022

Cited by 1 | Viewed by 3327

Abstract

Gurltia paralysans is a neglected and re-emerging metastrongyloid angio-neurotropic nematode causing severe chronic meningomyelitis in domestic cats (Felis catus) as well as in free-ranging small wild felids such as kodkods (Leopardus guigna), margays (Leopardus wiedii) and the [...] Read more.

Gurltia paralysans is a neglected and re-emerging metastrongyloid angio-neurotropic nematode causing severe chronic meningomyelitis in domestic cats (Felis catus) as well as in free-ranging small wild felids such as kodkods (Leopardus guigna), margays (Leopardus wiedii) and the northern tiger cat (Leopardus triginus) in South America. Within these definitive hosts (DH), adult males and females of G. paralysans parasitize the leptomeningeal veins of the subarachnoid space and/or the meningeal veins of spinal cord parenchyma, inducing vascular alterations. Feline gurltiosis has been associated with progressive thrombophlebitis of the meningeal veins, resulting in ambulatory paraparesis, paraplegia, ataxia, hindlimb proprioceptive deficit, uni- or bilateral hyperactive patellar reflexes, faecal and urinary incontinence, and tail paralysis. The complete life cycle of G. paralysans has not been elucidated yet, but most probably involves gastropods as obligate intermediate hosts (IH). In terms of epidemiology, G. paralysans infections in domestic and wild felids are scattered around various South American countries, with hyperendemic areas in southern parts of Chile. Etiological diagnosis of G. paralysans still represents a challenge for clinicians due to a lack of evidence of the excretion of either eggs or larvae in faeces or in other body fluids. Diagnosis is based on clinical neurological signs, imaging findings through computed tomography (CT), myelography, magnetic resonance imaging (MRI), and post mortem examination. Nonetheless, novel diagnostic tools have been developed, including semi-nested PCR for detecting circulating G. paralysans DNA in the cerebrospinal fluid, serum and blood samples as well as in serological diagnostic kits detecting parasite-derived antigens, but these need validation for routine usage. The hypothetical life cycle of G. paralysans is addressed in this article, including the exogenous stages (i.e., eggs, and first- (L1), second- (L2) and third-stage (L3) larvae) and obligate gastropod IH and/or paratenic hosts (PH), and we propose possible anatomical migration routes of infective L3 that reach the leptomeningeal veins in vivo. Finally, the pro-inflammatory endothelium- and leukocyte-derived innate immune reactions of the host against G. paralysans, which most likely result in thrombophlebitis and meningomyelitis, are briefly touched on. Full article

(This article belongs to the Section Parasitic Pathogens)

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Distribution of wild guiñas (syn. huiñas, kodkods, spotted tiger cat) in South America. (A) Adult specimen of a guiña (Leopardus guigna) (image reprinted with permission from © Joel Sartore/Photo Ark, 2022). (B) Geographic distribution of guiña in Chile (orange) and Argentina (yellow). (C) Scale representation of an adult guiña. Full article ">Figure 2
Proposed life cycle and migration pathways of Gurltia paralysans. (A) Cranial end of an adult specimen of G. paralysans. (B) Domestic cats (Felis catus) or wild felids (Leopardus spp.) acquire the L3 larvae by ingesting an infected obligate intermediate host (gastropods) or paratenic hosts (lizards, rodents, amphibians, birds or insects). Infective larvae penetrate the stomach and enter the hepatic portal system, and then the caudal vena cava and/or the azygous venous system. From these vein systems, the larvae migrate to the spinal cord via the intervertebral veins and the vertebral venous plexus. The larvae invade the veins of the subarachnoid space of the spinal cord, where they mature and lay eggs. It is still unknown on how domestic cats eliminate the eggs or the first-stage larvae (L1) into the environment, their further development into the L2 and L3 larval stages, or how the obligate intermediate hosts become infected with L1. AV: azygos vein; CV: caudal vena cava; IV: intervertebral veins; H: heart; L: liver; S: stomach; SC: spinal cord; VVP: vertebral venous plexus; L1: first-stage larvae; L2: second-stage larvae; L3: third-stage larvae. The inserted QR code shows a video of a G. paralysans-infected cat with clinical signs of paraparesis. Full article ">Figure 3
Microscopic view of Gurltia paralysans. (A) Cephalic end of the specimen showing a tooth at the anterior margin (scale bar: 50 µm). (B) Caudal end of a male, showing the small copulatory bursa (scale bar: 50 µm). (C) Higher magnification of a male caudal extremity, showing the spicules (arrow) (scale bar: 50 µm). Full article ">Figure 4
Macroscopic and microscopic lesions of the spinal cord in a Gurltia paralysans-infected cat. (A) Lumbar, sacral and caudal segments of the spinal cord showing severe and diffuse submeningeal vascular congestion. (B) Histopathological view of transverse sections of an adult of G. paralysans (arrows) inside a subarachnoid vein and vascular congestion (asterisk) in the spinal subarachnoid space; HE, 4 × (scale bar: 500 µm). (C) Histopathological section of the spinal cord parenchyma showing developing eggs of G. paralysans (arrow); HE, 40 × (scale bar: 50 µm). Full article ">Figure 5
Macroscopic view of the right lung with multiple haemorrhagic foci in a cat with feline gurltiosis. Full article ">

16 pages, 5623 KiB

Open AccessArticle

The Efficiency of Commercial Immunodiagnostic Assays for the Field Detection of Schistosoma japonicum Human Infections: A Meta-Analysis

by Zhongqiu Mei, Shan Lv, Liguang Tian, Wei Wang and Tiewu Jia

Pathogens 2022, 11(7), 791; https://doi.org/10.3390/pathogens11070791 - 13 Jul 2022

Cited by 2 | Viewed by 2309

Abstract

Although great strides have been achieved, schistosomiasis japonica remains a major public health concern in China. Immunodiagnostics have been widely accepted as the first choice in large-scale screening of Schistosoma japonicum human infections, and indirect hemagglutination test (IHA), enzyme-linked immunosorbent assay (ELISA), and [...] Read more.

Although great strides have been achieved, schistosomiasis japonica remains a major public health concern in China. Immunodiagnostics have been widely accepted as the first choice in large-scale screening of Schistosoma japonicum human infections, and indirect hemagglutination test (IHA), enzyme-linked immunosorbent assay (ELISA), and dipstick dye immunoassay (DDIA) are currently the three most common immunological tests for the diagnosis of S. japonicum human infections in China. This meta-analysis aimed to comprehensively assess the performance of IHA, ELISA, and DDIA for the field diagnosis of S. japonicum human infections. A total of 37 eligible publications were enrolled in the final analysis, including 29 Chinese publications and 8 English publications. No significant heterogeneities were detected among the studies reporting ELISA (I² = 88%, p < 0.05), IHA (I² = 95%, p < 0.05), or DDIA (I² = 84%, p < 0.05). DDIA showed the highest pooled sensitivity (90.8%, 95% CI: 84.6% to 94.7%) and IHA presented the highest pooled specificity for detection of S. japonicum human infections (71.6%, 95% CI: 65.9% to 76.7%). Summary receiver operating characteristic (SROC) curve analysis showed that IHA exhibited the highest area under the SROC curve (AUC) (0.88, 95% CI: 0.85 to 0.9), and ELISA presented the lowest AUC (0.85, 95% CI: 0.82 to 0.88). Deeks’ funnel plots indicated no publication bias. IHA presented the highest sensitivity in medium-endemicity regions and the highest specificity for diagnosis of S. japonicum human infections in low-endemicity regions, and ELISA showed the highest diagnostic sensitivity in high-endemicity regions and the highest specificity in medium-endemicity regions, while DDIA exhibited the highest diagnostic sensitivity in high-endemicity regions and the highest specificity in low-endemicity regions. IHA and DDIA presented a higher efficiency for the diagnosis of S. japonicum human infections in marshland and lake regions than in hilly and mountainous regions, while ELISA showed a comparable diagnostic sensitivity between in marshland and lake regions and hilly and mountainous regions (88.3% vs. 88.6%), and a higher specificity in marshland and lake regions than in hilly and mountainous regions (60% vs. 48%). Our meta-analysis demonstrates a comparable diagnostic accuracy of IHA, ELISA, and DDIA for S. japonicum human infections, and the diagnostic sensitivity and specificity of IHA, ELISA, and DDIA vary in types and infection prevalence of endemic regions. DDIA combined with IHA is recommended as a tool for screening chemotherapy targets and seroepidemiological surveys during the stage moving towards schistosomiasis elimination in China. Further studies to examine the effectiveness of combinations of two or three immunological tests for diagnosis of S. japonicum human infections are warranted. Full article

(This article belongs to the Special Issue Advanced Diagnosis of Schistosomiasis)

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Pathogens, Volume 11, Issue 7 (July 2022) – 109 articles

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