Multiple Infections Research Articles

Burden of multidrug-resistant bacteria among HIV-positive individuals in Ethiopia: A systematic review and meta-analysis.

Multidrug-resistant (MDR) bacteria are a significant cause of severe infections, particularly in human immunodeficiency virus (HIV)-positive individuals because of their weakened immunity. Since there was no previous pooled representative data regarding the MDR bacteria among HIV-positive individuals in Ethiopia, this systematic review and meta-analysis is required. This study was conducted based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. A literature search was performed using PubMed, Medline, EMBASE, Google Scholar, Hinari, Web of Science, Science Direct, and African Journals Online databases. Data were extracted using Microsoft Excel 2019 and analyzed using STATA version 11.0 software. A random-effects model was used to estimate the pooled effect size of outcome variables across studies with a 95% confidence interval. The I2 statistic was used to check for heterogeneity. The presence of publication bias was determined using a funnel plot and Egger's test with a p-value < 0.05 evidence of statistically significant bias. The pooled prevalence of MDR was 58.02% (95% CI: 46.32-69.73%) with high heterogeneity (I2 = 97.1%, (p < 0.001). In subgroup analysis, the highest multi-drug resistance was observed in the Oromia region (80.95%), patients with multiple infections (82.35%), and studies identified both Gram-positive and Gram-negative bacteria (61.45%). Furthermore, the pooled prevalence of MDR bacteria colonizing HIV-positive individuals was 48.76%. Regarding MDR species, Enterococci (77.41%) and Pseudomonas spp. (84.60%) were commonly identified in individuals with HIV infection. Our study indicates a high burden of MDR among HIV-positive individuals in Ethiopia. The Oromia region, HIV patients with multiple infections, Pseudomonas spp., and Enterococci showed the highest MDR in the subgroup analysis. Therefore, regional hospitals should implement strategies to tackle MDR such as vaccination program, appropriate use of antibiotics, and further study on the associated factors of MDR bacteria in HIV are required.

Eco-friendly synthesis of silver nanoparticles from peel and juice C. limon and their antiviral efficacy against HSV-1 and SARS-CoV-2

The growing threat of viral infections requires innovative therapeutic approaches to safeguard human health. Nanomaterials emerge as a promising solution to overcome the limitations associated with conventional therapies. The eco-friendly synthesis of silver nanoparticles (AgNPs) currently represents a method that guarantees antimicrobial efficacy, safety, and cost-effectiveness. This study explores the use of AgNPs derived from the peel (Lp-AgNPs) and juice (Lj-AgNPs) Citrus limon “Ovale di Sorrento”, cultivars of the Campania region. The antiviral potential was tested against viruses belonging to the Coronaviridae and Herpesviridae. AgNPs were synthesized by reduction method using silver nitrate solution mixed with aqueous extract of C. limon peel and juice. The formation of Lp-AgNPs and Lj-AgNPs was assessed using a UV–Vis spectrophotometer. The size, ζ-potential, concentration, and morphology of AgNPs were evaluated by dynamic light scattering (DLS), nanoparticle tracking analysis (NTA), and field emission-scanning electron microscopy (FE-SEM). Cytotoxicity was evaluated in a concentration range between 500 and 7.8 µg/mL on VERO-76 and HaCaT cells, with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium test bromide (MTT). Antiviral activity consisted of virus pre-treatment, co-treatment, cellular pre-treatment, and post-infection tests versus HSV-1 and SARS-CoV-2 at a multiplicity of infections (MOI) of 0.01. Plaque reduction assays and real-time PCR provided data on the antiviral potential of tested compounds. Lp-AgNPs and Lj-AgNPs exhibited spherical morphology with respective diameters of 60 and 92 nm with concentrations of 4.22 and 4.84 × 1010 particles/mL, respectively. The MTT data demonstrated minimal cytotoxicity, with 50 % cytotoxic concentrations (CC50) of Lp-AgNPs and Lj-AgNPs against VERO cells of 754.6 and 486.7 µg/mL. Similarly, CC50 values against HaCaT were 457.3 µg/mL for Lp-AgNPs and 339.6 µg/mL for Lj-AgNPs, respectively. In the virus pre-treatment assay, 90 % inhibitory concentrations of HSV-1 and SARS-CoV-2 were 8.54–135.04 µg/mL for Lp-AgNPs and 6.13–186.77 µg/mL for Lj-AgNPs, respectively. The molecular investigation confirmed the antiviral data, recording a reduction in the UL54 and UL27 genes for HSV-1 and in the Spike (S) gene for SARS-CoV-2, following AgNP exposure. The results of this study suggest that Lp-AgNPs and Lj-AgNPs derived from C. Limon could offer a valid ecological, natural, local and safe strategy against viral infections.

Good syndrome combined with multiple microbial pulmonary infections: case report and review of the literature.

Good syndrome (GS), a rare acquired immunodeficiency disorder characterized by thymoma and hypogammaglobulinemia, predisposes individuals to recurrent infections. This study reports a case of a 37-year-old male GS with multiple pulmonary infections and reviews relevant literature. The patient, with a history of thymoma resection, experienced multiple hospitalizations due to lung infections and neutropenia. The alveolar lavage fluid was detected by macro-genomic sequencing (NGS) to detect multiple pathogens, and targeted anti-infective and immunity-enhancing treatments led to improved symptoms and normal neutrophil counts. A literature review of 98 case reports from 2000 to 2023 was conducted, summarizing the associated diseases and pathogens in GS patients. Regular immunoglobulin monitoring in thymoma patients is essential for early GS diagnosis. When empirical antimicrobial therapy fails, mNGS for pathogen detection and targeted therapy are crucial, and regular IVIG injections can reduce infection rates in GS patients.

Infections and their prognostic significance before diagnosis of chronic lymphocytic leukemia, non-Hodgkin lymphoma, or multiple myeloma

BackgroundImmunodeficiency is a shared feature of B cell malignancies. The risk of infections and their prognostic significance after diagnosis are well characterized, but, conversely, less is known about prediagnostic infections in these domains.MethodsIn matched case-control analyzes, using Danish nationwide registers, we assessed the rate of prediagnostic infections in chronic lymphocytic leukemia (CLL), diffuse large B cell lymphoma (DLBCL), multiple myeloma (MM), follicular lymphoma (FL), marginal zone lymphoma (MZL), and lymphoplasmacytic lymphoma (LPL). Survival analyzes of data from clinical registers were then used to determine the effect of infections in the year preceding diagnosis on overall survival. To yield results for as many patients as possible, antimicrobial prescriptions were used as surrogates for infections.ResultsThe nationwide and clinical registers comprised 30,389 patients, accumulating 213,649 antimicrobial prescriptions, and 18,560 patients accumulating 107,268 prescriptions, respectively. The relative risk of infections was increased up to 15 years prior to diagnosis of malignancy and markedly increased in the year just prior to diagnosis. More than two antimicrobials within one year prior to diagnosis were associated with significantly shorter overall survival, independently of known prognostic factors.ConclusionPatients with B cell-derived malignancies exhibit marked immunodeficiency several years prior to diagnosis such that different disease subtypes demonstrate both overlapping and distinct trends in infection risk preceding diagnosis. Moreover, multiple infections within the year preceding diagnosis are independently associated with shorter overall survival for all the examined malignancies.

Multifunctional Chi-Ag@HMPB@CBD nanocomposites for eradicating multidrug-resistant bacteria and promoting diabetic damaged tissue repair

Injured tissues are vulnerable to polymicrobial infection, resulting in insufficient blood supply, excessive oxidative stress, and inflammation, seriously affecting skin regeneration. Therefore, there is an urgent need to develop effective strategies to treat multiple bacterial infections and damaged tissues and tissue repair for clinical treatment. Herein, we designed new nano-complexes of Chi-Ag@HMPB@CBD NPs via hollow mesoporous Prussian blue nanoparticles (HMPB NPs) loaded with cannabidiol (CBD) and chitosan-modified silver nanoparticles (Chi-Ag NPs) to treat multiple bacterial infected damaged tissues. In vitro experiments show that Chi-Ag@HMPB@CBD NPs synergizes with CBD and Chi-Ag NPs to effectively kill the mixture of Methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E. coli) via disrupting the cell membranes and established biofilms. Meanwhile, Chi-Ag@HMPB@CBD NPs can decompose H2O2 and increase O2 levels, thereby alleviating microenvironmental hypoxia and protecting cells from oxidative stress damage. Moreover, Chi-Ag@HMPB@CBD NPs can promote Human Umbilical Vein Endothelial Cells (HUVECs) proliferation, migration, and neovascularization by inhibiting Matrix Metalloprotein-9 (MMP-9) expression. In vivo experiments show that Chi-Ag@HMPB@CBD NPs exhibit high efficacy against mixed infections of E. coli and MRSA, accelerating damaged tissue repair via upregulating levels of Vascular Endothelial Growth Factor (VEGF) and platelet endothelial cell adhesion molecule-1 (CD31). Collectively, this study demonstrates that Chi-Ag@HMPB@CBD NPs are versatile nanomaterials with a great potential in clinical treatment of mixed bacterial-infected damaged tissue repair.

Metabolomic analysis reveals altered amino acid metabolism and mechanisms underlying Eimeria infection in laying hens

Avian coccidiosis, caused by Eimeria spp, is a devastating disease in laying hens. Previous studies have suggested that amino acids may be involved in Eimeria infection of broiler chickens. However, their metabolic features in laying hens, as well as the effect of multiple Eimeria species challenges on poultry hosts have not been elucidated yet. Here, a targeted metabolomics approach was employed to identify altered amino acid metabolism and mechanisms in laying hens with multiple Eimeria species challenges. Laying hens, Hy-Line W-36 aged 25 wk, were randomly assigned to a control group and groups inoculated with varying levels of mixed Eimeria species (E. maxima, E. tenella, and E. acervulina). Serum samples from each group were collected at 6 d and 14 d of postinoculation (6 and 14 DPI) for metabolite profiling. Metabolomic analysis revealed notable metabolic variations between control and infected groups, especially at 6 DPI stage. Varying levels of Eimeria dosages did not show a significant metabolic difference, and metabolites were sensitive to low-level infection. With statistical analysis, differentially expressed compounds (3-methylhistidine, alanine, aspartate, lysine, asparagine, methionine, ornithine, and tryptophan) were selected, and their metabolic network was identified by pathway enrichment analysis. In the network, the lysine biosynthesis pathway was upregulated, while the arginine and proline metabolic pathway was downregulated under infection. Other pathways showed complex patterns of metabolic relationships. Based on the results, biological implications of metabolic changes were elucidated and discussed. Last, the results were further confirmed with our previous study (phenotype and gene expression results) using the same set of samples. Our finding provides in-depth information on altered amino acid metabolism and mechanisms in laying hens upon multiple Eimeria species infection.

Chemical optimization and derivatization of micrococcin p2 to target multiple bacterial infections: new antibiotics from thiopeptides.

Antimicrobial resistance poses a significant threat to humanity, and the development of new antibiotics is urgently needed. Our research has focused on thiopeptide antibiotics such as micrococcin P2 (MP2) and derivatives thereof as new anti-infective agents. Thiopeptides are sulfur-rich, structurally complex substances that exhibit potent activity against Gram-positive pathogens and Mycobacteria species, including clinically resistant strains. The clinical development of thiopeptides has been hampered by the lack of efficient synthetic platforms to conduct detailed structure-activity relationship studies of these natural products. The present contribution touches upon efficient synthetic routes to MP2 that laid the groundwork for clinical translation. The medicinal chemistry campaign on MP2 has been guided by computational molecular dynamic simulations and parallel investigations to improve drug-like properties, such as enhancing the aqueous solubility and optimizing antibacterial activity. Such endeavors have enabled identification of promising lead compounds, AJ-037 and AJ-206, against Mycobacterium avium complex (MAC). Extensive in vitro studies revealed that these compounds exert potent activity against MAC species, a subspecies of non-tuberculous mycobacteria (NTM) that proliferate inside macrophages. Two additional pre-clinical candidates have been identified: AJ-024, for the treatment of Clostridioides difficile infections, and AJ-147, for methicillin-resistant Staphylococcus aureus impetigo. Both compounds compare quite favorably with current first-line treatments. In particular, the ability of AJ-147 to downregulate pro-inflammatory cytokines adds a valuable dimension to its clinical use. In light of above, these new thiopeptide derivatives are well-poised for further clinical development.

Patterns of single and multiple HPV infections in female: A systematic review and meta-analysis

BackgroundData on the patterns of single and multiple HPV infections are largely limited to small size studies, and the regional difference have not been systematically examined. MethodsA literature search was conducted using PubMed, Embase, and Web of Science databases up to Sept 22, 2023. The pooled prevalence of HPV infection were calculated using random-effects meta-analysis. Subgroup analysis was used to explore the heterogeneity, and publication bias was evaluated by Egger's test and Begg's test. ResultsThere were 121 studies included with 1,682,422 participants. Globally, the most common genotypes of single HPV infection were HPV16 (7.05 %), 18 (1.94 %), 52 (1.93 %), 58 (1.68 %), and 31 (1.53 %), as well as HPV 16 (4.91 %), 31 (2.68 %), 52 (2.20 %), 51 (1.99 %), and 18 (1.96 %) in multiple HPV infections. Apart from HPV16 and 18, HPV52 and 58 were common in Asia, HPV31 and 51 was in Europe, North and South America, and HPV35 and 45 were in Africa. The prevalence of HPV infection among different age groups (<30, 30–50, >50 years age groups) was 20.93 %, 16.27 %, and 18.69 %, respectively. The single HPV infection prevalence in the No-ILs, LSILs, HSILs, and cervical cancer groups were 16.17 %, 51.60 %, 57.12 %, and 62.88 %, respectively, as well as in multiple infections were 5.09 %, 30.93 %, 32.86 %, and 21.26 % ConclusionDeveloping local HPV vaccines is necessary based on the HPV infection pattern. It is essential to educate young women to get vaccinated and encourage elderly women to have regular cervical cancer screenings to reduce the danger of cervical cancer.

CFTR Inhibitors Display Antiviral Activity against Herpes Simplex Virus.

The cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-dependent Cl- channel, is closely associated with multiple pathogen infections, such as SARS-CoV-2. However, whether the function of the CFTR is involved in herpes simplex virus (HSV) infection has not been reported. To evaluate the association of CFTR activity with HSV infection, the antiviral effect of CFTR inhibitors in epithelial cells and HSV-infected mice was tested in this study. The data showed that treatment with CFTR inhibitors in different concentrations, Glyh-101 (5-20 μM), CFTRi-172 (5-20 μM) and IOWH-032 (5-20 μM), or the gene silence of the CFTR could suppress herpes simplex virus 1 (HSV-1) and herpes simplex virus 2 (HSV-2) replication in human HaCaT keratinocytes cells, and that a CFTR inhibitor, Glyh-101 (10-20 μM), protected mice from HSV-1 and HSV-2 infection. Intracellular Cl- concentration ([Cl-]i) was decreased after HSV infection via the activation of adenylyl cyclase (AC)-cAMP signaling pathways. CFTR inhibitors (20 μM) increased the reduced [Cl-]i caused by HSV infection in host epithelial cells. Additionally, CFTR inhibitors reduced the activity and phosphorylation of SGK1 in infected cells and tissues (from the eye and vagina). Our study found that CFTR inhibitors can effectively suppress HSV-1 and HSV-2 infection, revealing a previously unknown role of CFTR inhibitors in HSV infection and suggesting new perspectives on the mechanisms governing HSV infection in host epithelial cells, as well as leading to potential novel treatments.

Association of Methylated DNA Markers with High-Risk HPV Infections in Oral Site and Precancer Anal Lesions in HIV-Positive MSM.

Human papillomavirus (HPV) infection is linked to several cancers, including anal and oral cancers. The incidence of anal cancer is particularly high among HIV-positive men who have sex with men (MSM). DNA methylation markers have shown promise as biomarkers for identifying precancerous lesions and cancer in HPV-infected individuals. The aim of this study was to investigate the correlation of DNA methylation with HPV infection in oral samples and the correlation of DNA methylation with lesion degree in the anal samples of HIV-positive MSM. This study investigated DNA methylation in oral and anal samples from HIV-positive MSM at the National Institute for Infectious Diseases (INMI) in Rome, Italy. Exfoliated oral epithelial cells and anal samples were collected and analyzed for 28 HPV genotypes using the Allplex 28 HPV assay. DNA methylation was assessed with the PrecursorM+ kit for oral samples and the AnoGyn kit for anal samples, focusing on the promoter regions of specific genes. The study included 63 participants, with a median age of 49 and a median CD4+ count of 705 cells/µL. The oral samples showed HPV16 as the most common type, with 22% testing positive for DNA methylation. The anal samples exhibited HPV-related methylation changes linked to cytological lesions, with a 30% increase in the observed ddCt ratio. Significant differences were found in both ASCL1 and ZNF582 genes, particularly for HSILvsNILM and HSILvsLSIL lesions. Of the samples with an increased ddCt ratio, 80% were from patients over 35 years old, and multiple HPV infections were common. DNA methylation markers could be valuable in identifying high-risk HPV infections in oral samples and detecting potential precancerous lesions in anal samples. These markers may enhance the early detection and prevention strategies for HPV-related cancers in high-risk populations, with follow-up data indicating potential for monitoring lesion progression.

Asymptomatic Malaria Cases and Plasmodium Species among BaAka Pygmies in Central Africa.

Malaria is a significant health problem in Africa, primarily due to the Plasmodium falciparum species, but this is not the only etiological factor responsible for malaria on the continent. The goal of the present research was to describe asymptomatic malaria cases and to identify Plasmodium species responsible for malaria in the BaAka Pygmies, inhabitants of the Central African Republic (CAR). Screening was realised in the period of August-September 2021 among 308 people, including 74 children and 234 adults reporting to a healthcare facility in Monasao (southwest CAR), an area inhabited by a semi-nomadic tribe of BaAka Pygmies. The study consisted of two phases. Phase I, which was conducted in Africa, consisted of performing malaria rapid diagnostic tests (mRDTs), taking haemoglobin measurements and collecting blood samples onto Whatman FTA cards for molecular diagnostics. Phase II, which was conducted in Poland, involved molecular tests (RT-PCR) to confirm or rule out malaria infections and to identify Plasmodium species responsible for the infections. mRDTs detected Plasmodium infections in 50.3% of children and 17.1% of adults participating in the study, whereas RT-PCR assays yielded positive results for 59.5% children and 28.6% adults. Molecular tests detected multiple Plasmodium falciparum infections but also three infections with P. malariae, three with P. ovale and one with P. vivax. The obtained results have confirmed numerous asymptomatic Plasmodium infections among the BaAka Pygmies. The rates of asymptomatic malaria cases in adults were twice as high as those in children, which may be indicative of the gradual acquisition of protective immunity with age. The study findings have also demonstrated that although most cases of malaria in Africa are caused by P. falciparum, three other species are also present in the region.

Preliminary Study on Ecto and Endoparasites of the Black Rats (Rattus Rattus) in Benghazi City

Forty-five adult trapped black rats (Rattus rattus), 24 males and 21 females, were dissected and examined in this study for ecto and endoparasites. Three ectoparasites were detected, namely, the flea (Xenopsallya cheopis), mites (Ornithonyssus bacoti), and tick (Ixodes spp) with 48.9%, 13.3%, and 2.2% respectively. Meanwhile, many species of endoparasites were: Entamoeba histolytica/dispar (73.3%), Entamoeba coli (31.3%), Blastocystis hominus (8.9%), Giardia lamblia (15.6), Chilomastix sp. (4.4%), a coccidia sp. (13.3%), Hymenolips nana (2.2%), Hymenolips diminuta (44.4%), and Taenia taeniaformis (6.7%), and Moniliformis moniliformis (2.2%). Of all examined rats, 97.8 % (44/45) were found to harbor at least one parasite species with no significant differences between body weight, body length, and gender of the host with the prevalence of parasite species. Multiple infections occurred at six species per rat with different combinations of parasite infections. These results are the first records of the parasitic fauna of R. rattus in the Benghazi area.

HPV specificity and multiple infections and association with cervical cytology in Chongqing, China: a cross-sectional study

BackgroundIt is important to assess the relationship between specific HPV genotype or multiple infection and cervical cytology. The protection provided by the HPV vaccine is type-specific, and the epidemiology feature of coinfections needs to be investigated. The aim is to provide baseline information for developing HPV vaccination and management of HPV-positive populations in the region.MethodsA total of 3649 HPV-positive women were collected from 25,572 women who underwent 15 HR-HPV genotypes and ThinPrep cytologic test (TCT) results. Logistic regression was used to determine the correlation between the risk of cytology abnormalities and specific HPV infection. We calculated odds ratios (ORs) to assess coinfection patterns for the common two-type HPV infections. chi-squared test was used to estimate the relationship between single or multiple HPV (divided into species groups) infection and cytology results.ResultsThe results showed there was a positive correlation between HPV16 (OR = 4.742; 95% CI 3.063–7.342) and HPV33 (OR = 4.361; 95% CI 2.307–8.243) infection and HSIL positive. There was a positive correlation between HPV66 (OR = 2.445; 95% CI 1.579–3.787), HPV51 (OR = 1.651; 95% CI 1.086–2.510) and HPV58(OR = 1.661; 95% CI 1.166–2.366) infection and LSIL. Multiple HPV infections with α9 species (OR = 1.995; 95% CI 1.101–3.616) were associated with a higher risk of high-grade intraepithelial lesions (HSIL) compared with single HPV infection. There were positive correlations between HPV66 and HPV56 (α6) (OR = 3.321; 95% CI 2.329–4.735) and HPV39 and HPV68 (α7). (OR = 1.677; 95% CI 1.127–2.495). There were negative correlations between HPV52, 58, 16 and the other HPV gene subtypes.ConclusionHPV33 may be equally managed with HPV16. The management of multiple infections with α9 may be strengthened. The 9-valent vaccine may provide better protection for the population in Chongqing currently. The development of future vaccines against HPV51 and HPV66 may be considered in this region.

Influence of biosecurity on the occurrence of various enteric viruses in broiler flocks

ABSTRACT Common enteric viruses affecting commercial broiler flocks include fowl adenovirus (FAdV), chicken parvovirus (ChPV), chicken astrovirus (CAstV), avian reovirus (ARV) and avian rotavirus (AvRV). To investigate their prevalence and to identify single and multiple infections we collected intestinal samples from 49 Austrian broiler flocks during necropsy of dead-on-farm birds twice during a production cycle (7–14 days and 28–35 days). Altogether, up to three consecutive clinically healthy flocks without signs of gastrointestinal disease were sampled from 17 different farms. Samples were analysed using virus isolation and PCR/RT–PCR methods. Virus prevalence was correlated with production data and on-farm biosecurity and management practices. Overall, ARV (75%) was most commonly detected in the flocks, followed by CAstV (61%), ChPV (61%), FAdV (57%) and AvRV (8%). Only in three (6%) flocks were none of the investigated enteric viruses detected. Flock infection profiles were very heterogeneous and included individual detection of the investigated viruses as well as different combinations thereof (up to all five investigated viruses). Even in the absence of clinical diarrhoea and/or macroscopic intestinal lesions, statistical analysis confirmed that the number of viruses detected had a significant economic impact characterised by poor weight gain and increased mortality, particularly due to the presence of FAdV, CAstV and/or ARV. Furthermore, the use of barn-specific clothing and/or footbaths as well as regular vermin control, resulted in lower prevalence of enteric viruses in the flocks studied. This highlights the importance of common biosecurity measures in poultry production to prevent economic losses. RESEARCH HIGHLIGHTS Detection timepoints and patterns indicate horizontal introduction of various enteric viruses. Flock infection profiles were very heterogeneous; no dominating virus profile. Broiler production was negatively affected by the number of enteric viruses detected. Common biosecurity measures had a significant negative effect on virus prevalence.

Prevalence and molecular analysis of some important viruses in honey bee colonies in Türkiye: the status of multiple infections.

In this study, seven bee viruses of significant importance for bee health in Türkiye were investigated using one-step RT-PCR. For this purpose, larvae from 1183 hives and adult bees from 1196 hives were sampled from 400 apiaries in 40 provinces. The prevalence of viral infections in hives was as follows: acute bee paralysis virus (ABPV), 6.4%; black queen cell virus (BQCV), 77%; chronic bee paralysis virus (CBPV), 3.2%; deformed wing virus (DWV), 63.8%; Israel acute bee paralysis virus (IAPV), 7%; Kashmir bee virus (KBV), 2.7%; sacbrood virus (SBV), 49.7%. Moreover, 50 different combinations of viral infections were identified in the hives. While dual infections (36.1%) were the most common in hives, triple infections with BQCV, DWV, and SBV were found to have the highest prevalence (22.1%). At least one viral infection was detected in all of the apiaries tested. Phylogenetic analysis showed that the isolates from this study generally exhibited the highest similarity to previously reported Turkish isolates. When similarity ratios and the locations and types of amino acid mutations were analyzed, it was observed that the isolates from our study exhibited high similarity to isolates from various countries, including China, the United Kingdom, Syria, and Germany.

Pathogenicity of Duck Adenovirus Type 3 in Chickens.

Duck adenovirus Type 3 (DAdV-3) severely affects the health of ducks; however, its pathogenicity in chickens remains unknown. The objectives of this study were to evaluate the pathogenicity and major pathological changes caused by DAdV-3 in chickens. Viral DNA was extracted from the liver of the Muscovy duck, and the fiber-2 and hexon fragments of DAdV-3 were amplified through polymerase chain reaction (PCR). The evolutionary tree revealed that the isolated virus belonged to DAdV-3, and it was named HE-AN-2022. The mortality rate of chicks that received inoculation with DAdV-3 subcutaneously via the neck was 100%, while the mortality rate for eye-nose drop inoculation was correlated with the numbers of infection, with 26.7% of chicks dying as a result of exposure to multiple infections. The main symptoms exhibited prior to death were hepatitis-hydropericardium syndrome (HHS), ulceration of the glandular stomach, and a swollen bursa with petechial hemorrhages. A histopathological examination revealed swelling, necrosis, lymphocyte infiltration, and basophilic inclusion bodies in multiple organs. Meanwhile, the results of quantitative real-time PCR (qPCR) demonstrated that DAdV-3 could affect most of the organs in chickens, with the gizzard, glandular stomach, bursa, spleen, and liver being the most susceptible to infection. The surviving chicks had extremely high antibody levels. After the chickens were infected with DAdV-3 derived from Muscovy ducks, no amino acid mutation was observed in the major mutation regions of the virus, which were ORF19B, ORF66, and ORF67. On the basis of our findings, we concluded that DAdV-3 infection is possible in chickens, and that it causes classic HHS with ulceration of the glandular stomach and a swollen bursa with petechial hemorrhages, leading to high mortality in chickens. The major variation domains did not change in Muscovy ducks or in chickens after infection. This is the first study to report DAdV-3 in chickens, providing a new basis for preventing and controlling this virus.

Bacilli load in PTB- intestinal helminths co-infected and PTB non -infected patients at selected public health facilities in Jimma zone, Oromia, Ethiopia: comparative cross-sectional study

BackgroundTuberculosis (TB) and intestinal helminths are diseases that pose a dual burden on public health in low-income countries. Previous studies have shown that helminths can affect the shedding of bacteria or the bacterial load in the sputum of active TB patients. However, there is limited information on bacterial load in TB patients with helminth infections.ObjectiveThis study aimed to compare bacterial load in helminths-infected and non-infected pulmonary tuberculosis patients at selected public health facilities in Jimma zone, Oromia, Ethiopia.MethodsThe study was conducted in Jimma Zone, Oromia, Ethiopia. A facility-based comparative cross-sectional study was employed from August 01, 2020, to January 2021. A total of 124 (55 intestinal helminths-infected and 69 non-infected) newly diagnosed smear-positive pulmonary tuberculosis (PTB) patients were included in the study. A convenience sampling technique was employed to recruit study participants, and a semi-structured questionnaire was used to collect data regarding socio-demographic characteristics and possible risk factors for intestinal helminths co-infection. Stool examination was performed using both wet mount and Kato Katz technique. Additionally, weight and height measurements, sputum, and blood samples were taken to determine body mass index, bacilli load, and diabetic mellitus, respectively. Data were entered into Epi-Data software version 3.1 and analyzed using Statistical Packages for Social Sciences (SPSS) Version 25. A statistically significant difference was defined as a P-value of less than 0.05.ResultsIntestinal helminths reduced bacilli load 3 times more than intestinal helminths non-infected PTB (AOR = 3.44; 95% CI; 1.52, 7.79; P = 0.003) However, diabetes mellitus, HIV, drinking alcohol and cigarette smoking were not associated with bacilli load. The rate of co-infection TB with intestinal helminths was 44%. The three most prevalent parasites detected were Trichuris trichiura 29 (66%), hookworm 19 (43%), and Ascaris lumbricoides 11(25%)). Among co-infected patients about 36 (81.8%) had a single parasite infection, and 19 (43.2%) had multiple infections. A body mass index < 18.5 (AOR = 3.26; 95% CI; 1.25, 8.56;P = 0.016) and untrimmed fingernail status (AOR = 3.63; 95%CI;1.32,9.93;P = 0.012) were significantly associated with PTB- intestinal helminth –co-infection.ConclusionHelminth infection was associated with a lower bacilli load compared to helmenths non-infected PTB. The rate of co-infection TB with intestinal helminths was 44%. Trichuris trichiura was the most prevalent helminth. Untrimmed fingernail and a body mass index were associated with PTB-intestinal helminth co-infection.

MOIRE: A software package for the estimation of allele frequencies and effective multiplicity of infection from polyallelic data.

Malaria parasite genetic data can provide insight into parasite phenotypes, evolution, and transmission. However, estimating key parameters such as allele frequencies, multiplicity of infection (MOI), and within-host relatedness from genetic data has been challenging, particularly in the presence of multiple related coinfecting strains. Existing methods often rely on single nucleotide polymorphism (SNP) data and do not account for within-host relatedness. In this study, we introduce a Bayesian approach called MOIRE (Multiplicity Of Infection and allele frequency REcovery), designed to estimate allele frequencies, MOI, and within-host relatedness from genetic data subject to experimental error. Importantly, MOIRE is flexible in accommodating both polyallelic and SNP data, making it adaptable to diverse genotyping panels. We also introduce a novel metric, the effective MOI (eMOI), which integrates MOI and within-host relatedness, providing a robust and interpretable measure of genetic diversity. Using extensive simulations and real-world data from a malaria study in Namibia, we demonstrate the superior performance of MOIRE over naive estimation methods, accurately estimating MOI up to 7 with moderate sized panels of diverse loci (e.g. microhaplotypes). MOIRE also revealed substantial heterogeneity in population mean MOI and mean relatedness across health districts in Namibia, suggesting detectable differences in transmission dynamics. Notably, eMOI emerges as a portable metric of within-host diversity, facilitating meaningful comparisons across settings, even when allele frequencies or genotyping panels are different. MOIRE represents an important addition to the analysis toolkit for malaria population dynamics. Compared to existing software, MOIRE enhances the accuracy of parameter estimation and enables more comprehensive insights into within-host diversity and population structure. Additionally, MOIRE's adaptability to diverse data sources and potential for future improvements make it a valuable asset for research on malaria and other organisms, such as other eukaryotic pathogens. MOIRE is available as an R package at https://eppicenter.github.io/moire/.

Single-cell analysis of VACV infection reveals pathogen-driven timing of early and late phases and host-limited dynamics of virus production.

The extent and origin of variation in the replication dynamics of complex DNA viruses is not well-defined. Here, we investigate the vaccinia virus (VACV) infection cycle at the single-cell level, quantifying the temporal dynamics of early and post(dna)-replicative phase gene expression across thousands of infections. We found that viral factors determine the initiation time of these phases, and this is influenced by the multiplicity of infection (MOI). In contrast, virus production dynamics are largely constrained by the host cell. Additionally, between-cell variability in infection start time and virus production rate were strongly influenced by MOI, providing evidence for cooperativity between infecting virions. Blocking programmed cell death by pan-caspase inhibition increased infection frequency but not virus production at the population level due to a concurrent attenuation of per-cell virus yield, suggesting a dual role for caspase signaling in VACV infection. Our findings provide key insights into the pivotal factors influencing heterogeneity in the infection cycle of a large DNA virus at the single-cell level.

Virus diseases limiting greenhouses and open field production of cucurbits in Antalya province

The Mediterranean region in Türkiye is noted for the production of cucurbits and other vegetables. As such, the aim of this study was to sample symptomatic cucurbits crops from open fields and greenhouses where there is active cultivation of these crops. Young shoot, leaves, and fruits exhibiting virus-like symptoms (yellowing, mosaic, necrotic) were collected. Out of 968 plant samples collected and tested through RT-PCR and PCR, 949 were discovered to be infected with several viruses. The identified virus diseases included ZYMV, WMV, PRSV, SqMV, CGMV, CYSDV, BPYV, CABYV, ToLCNDV and CMV. In terms of hosts exhibiting a high incidence of virus infections, cucumber (363 samples), squash (277 samples), melon (201 samples), and watermelon (108 samples) emerged as the top four hosts. Additionally, viruses with notable high incidences in the collected samples, as recorded through molecular testing in decreasing order included ZYMV at 28.1%, CYSDV at 15.5%, and WMV at 14.4%. Also, 90% of samples collected from open fields had single or multiple infection. In contrast, 26.4% of samples from greenhouses exhibited mosaic symptoms and 74.6% showed yellowing symptoms. Notably, MNSV and BPYV, were detected in these samples. The samples also exhibiting mixed infections predominantly displayed mosaic symptoms, including mixed infections such as ZYMV with WMV, ZYMV with CMV, CMV with WMV, and CMV with PRSV. In contrast, samples obtained from open fields showed a higher prevalence of yellowing symptoms, such as ToLCNDV with CMV, ToLCNDV with ZYMV, ZYMV with CMV with CYSDV, and ZYMV with CMV with CVYV.