Post Infection Research Articles

Prevalence of Post Caesarean Wound Infections and Associated Factors among Mothers Delivering from Kampala International University Teaching Hospital

Caesarean section simply is an operative technique by which a fetus is delivered through an abdominal and uterine incisions after a minimum period of 28 weeks of amenorrhea, whilst a post caesarean section wound infection (PCWI) is an infection that occurs after a C- section, usually due to a bacterial infection in the surgical incision site. The purpose of this study was to determine the prevalence of post caesarean wound infections among mothers who delivered from KIU-TH, and to identify common factors associated with post caesarean wound infections among these mothers. Descriptive statistics were used to characterize and highlight the difference among the post-caesarean patients. Data was entered and analyzed using SPSS version 16. The analyzed data was then presented using tables and charts. The results indicate that the prevalence of post-caesarean wound infection at Kampala International University-Teaching Hospital is still low (3.2%) but may increase if the steps are not taken to initiate its prevention and eradication. The results indicate that post-caesarean wound infection is positively and significantly associated with age (X2 = 10.747, P<0.05), the cause/reason of caesarean section (X2 = 62.000, P<0.05), color of liquor (X2 =45.983, P<0.05), duration of labor (X2 =10.067, P< 0.05) and amount of blood lost (X2 = 3.844, P<0.05). The prevalence of PCWIs at KIU-TH was 3.2% and the factors associated were the reason for caesarean section, color of liquor, duration of labor, amount of blood lost. Based on the results and the findings, the study recommends that recognition of the consequences and the building of the strategies to prevent and treat Post-caesarean wound infections at Kampala international university-teaching hospital is essential for reducing post- caesarean maternal morbidity and mortality. In addition, the study also recommends the recognition of the associated risk factors particularly the modifiable and their modification to reduce the occurrence of Post-caesarean wound infection. Keywords: Prevalence, Post Caesarean, Wound Infections and Mothers Delivering

Routine blood test markers for predicting liver disease post HBV infection: precision pathology and pattern recognition.

Early stages of hepatitis B virus (HBV) infection usually involve inflammation of the liver. Patients with chronic infection have an increased risk of progressive liver fibrosis, cirrhosis, and life-threatening clinical complications of end-stage hepatocellular carcinoma (HCC). Early diagnosis of hepatic fibrosis and timely clinical management are critical to controlling disease progression and decreasing the burden of end-stage liver cancer. Fibrosis staging, through its current gold standard, liver biopsy, improves patient outcomes, but the clinical procedure is invasive with unpleasant post-procedural complications. Routine blood test markers offer promising diagnostic potential for early detection of liver disease without biopsy. There is a plethora of candidate routine blood test markers that have gone through phases of biomarker validation and have shown great promise, but their current limitations include a predictive ability that is limited to only a few stages of fibrosis. However, the advent of machine learning, notably pattern recognition, presents an opportunity to refine blood-based non-invasive models of hepatic fibrosis in the future. In this review, we highlight the current landscape of routine blood-based non-invasive models of hepatic fibrosis, and appraise the potential application of machine learning (pattern recognition) algorithms to refining these models and optimising clinical predictions of HBV-associated liver disease. Machine learning via pattern recognition algorithms takes data analytics to a new realm, and offers the opportunity for enhanced multi-marker fibrosis stage prediction using pathology profile that leverages information across patient routine blood tests.

Regulatory effects of potassium channel blockers on potassium channel genes upon nervous necrosis virus infection in sevenband grouper Hyporthodus septumfasciatus

Ion channels in fishes regulate the flow of important ions that play an active role in the excitation and transmission of impulses through neuronal cells. Specific housekeeping genes translates into proteins and selectively permeabilize and facilitate ion crossover transmissions. Potassium (K+) channels play a crucial role in a wide range of functions such as cell volume regulation, hormone secretion, synaptic transmission and muscle contraction. The dysfunction of ion channels result in channelopathies, which hinder critical cellular activities. Recent studies have indicated that viral pathogens tend to regulate cellular ion channels for entry into host cells. Hence, the present study aimed to elucidate the role of K+ channels during nervous necrosis virus (NNV) infections in the sevenband grouper (Hyporthodus septumfasciatus). Real-time PCR with the standardized potassium genes revealed the downregulation of potassium two pore domain channel subfamily member - KCNK10, KCNK9, KCNK2, and KCNK1 genes post infection at both 17 °C and 25 °C whereas an upregulation was noted in the case of gill tissues. SMART analysis revealed a transmembrane region in all genes. Multiple sequence alignment using MultAlin and phylogenetic analysis revealed true homology of potassium genes with other higher vertebrates. In vitro and in vivo challenge study of NNV using Tetra ethyl ammonium (TEA) as potential drug showed inverse relation to that of viral replication and a corresponding downregulation of K+ channel gene expression was observed which was further confirmed by an immunofluorescence assay. These findings indicate that K+ channels play a crucial role during viral infection. Moreover, the observed downregulation can be related to rapid endocytosis resulting from recycling endosomes during a viral infection. Hence, further studies are warranted to better understand the role of K+ channel genes during NNV infection.

Effects of Anticoccidial Vaccination and Taraxacum officinale Extract on the Growth Performance, Biochemical Parameters, Immunity, and Intestinal Morphology of Eimeria-Challenged Chickens.

A total of 160 Ross 308 male chickens were used in a 2 × 2 factorial design to examine the effects of anticoccidial vaccination (ACV; lack or 1× dose recommended by the manufacturer) and dietary supplementation with Taraxacum officinale (dandelion) extract (DE; with or without) on growth performance, immunity, biochemical parameters, and intestinal morphology in broiler chickens challenged with Eimeria spp. At 20 days of age, all birds were challenged with a 25× dose of ACV, including Eimeria acervulina, E. maxima, E. mitis, and E. tenella. No interaction between ACV and DE was observed in terms of growth performance. Vaccinated birds showed increased feed intake (FI) and feed conversion ratio (FCR) during the 11-20 day period. Meanwhile, DE supplementation led to decreased FI and body weight gain (BWG) during the 1-10 day period. ACV effectively induced immunity against Eimeria, as evidenced by reduced oocyst shedding and less intestinal lesions, decreased levels of pro-inflammatory interleukin-6, and improved BWG during both the post infection (PI) period (21-35 days) and the entire growth period. DE supplementation lowered FCR and increased BWG during the 35-42 day period, increased the concentration of butyric acid in the cecal digesta, and lowered oocyst shedding PI. In vaccinated birds, DE elevated levels of plasma total protein and immunoglobulin M, and influenced tight junction proteins zonula occludens-1 and claudin-3, indicating a more robust epithelial barrier. DE also lowered alanine aminotransferase activity in unvaccinated birds. Both ACV and DE independently improved intestinal morphology in the jejunum, decreasing crypt depth and increasing the villus height-to-crypt ratio. These findings suggest that both ACV and DE could be effective strategies for managing coccidiosis in broiler chickens.

Relative biochemical and physiological docking of cucumber varieties for supporting innate immunity against Podosphaera xanthii

Powdery mildew in cucumber is caused by the Podosphaera xanthii. No strategy for improving disease resistance can be successful in the absence of thorough insights into the physiological and biochemical responses of cucumber plants to powdery mildew. Therefore, a field experiment was executed to evaluate five commercial cucumber varieties (V1: Dynasty, V2: Long green, V3:Desi Kheera, V4:Thamin II, V5:Cucumber 363) for their inherent immunity to powdery mildew. Upon inoculating cucumber plants with Podosphaera xanthii, we noted differential responses among the varieties. Compared to other varieties, V1 and V2 showed higher values (P ≤ 0.05) for chlorophyll-a under control and pathogen-attacked plants respectively. The minimum value of anthocyanin content (−53.73%) was recorded in V3 as compared to other varieties post pathogen infection. All pathogen-infected cucumber varieties showed a considerable (P ≤ 0.05) loss in flavonoid content except V2. The maximum destruction for Phenolics under powdery mildew (179%) were recorded in V4, whereas V1 exhibited maximum phenolic content under control conditions. In pathogen-infected plants, the minimum AsA was recorded in V5 as compared to all other varieties. Pathogen invasion impacted significantly (P ≤ 0.05) the activity of superoxide dismutase (SOD). Besides, cucumber plants after pathogen inoculation resulted in a considerable (P ≤ 0.05) increase of peroxidase (POD) activity in V1 (5.02%), V2 (7.5%), and V3 (11%) in contrast to V4. Our results confirmed that cucumber varieties perform differently, which was brought on by distinct metabolic and physiological modifications that have an impact on growth and development. The changes in different attributes were correlated with cucumber resistance against powdery mildew. The results would help us fully harness the potential of these varieties to trigger disease management initiatives and defense responses.

Administration of Ethanolic Extract of Spinacia oleracea Rich in Omega-3 Improves Oxidative Stress and Goblet Cells in Broiler Chickens Infected with Eimeria tenella.

This study investigated the anticoccidial activity of spinach (Spinacia oleracea) whole-plant extract against Eimeria tenella, both in vitro and in vivo. For this purpose, one hundred 8-day-old broiler chicks of both sexes were divided into four groups (n = 25 in each group). Chicks in the first group served as the negative control (non-treated-non-infected). Chicks in the second group were challenged at 18 days old with 5 × 104E. tenella sporulated oocysts. The third group was challenged with 5 × 104 sporulated E. tenella oocysts at 18 days old after receiving spinach extract at a dose of 50 mg/kg at 8 days old. The fourth group received 0.2 mg/kg diclazuril (Coxiril® 0.2%) in their diet two days before being orally infected with 5 × 104 sporulated E. tenella oocysts and this continued till day 10 post-infection (PI). The growth performance, clinical symptoms, oocyst shedding, histological findings, and biochemical parameters were used to evaluate the efficacy on day 8 PI when the infection was at its peak. A gas chromatography examination revealed that omega-3 fatty acids were the main constituents of the spinach extract, followed by oleic acid, palmitic acid, and phytol, with amounts of 23.37%, 17.53%, 11.26%, and 7.97%, respectively. The in vitro investigation revealed that the spinach extract at concentrations of 10% and 5% inhibited the oocyst sporulation by 52.1% and 45.1%, respectively. The 5% concentration was selected for the in vivo trial based on the results of the in vitro study. The infected-untreated group showed high levels of OPG; lower body weight; a greater number of parasite stages; few goblet cells; decreased SOD, CAT, and GPX levels; and increased MDA and NO levels. The spinach-treated group, on the other hand, showed a significant decrease in oocyst output per gram of feces (OPG), increased body weight, decreased parasitic stages, and a nearly normal number of goblet cells. Additionally, it reduced malondialdehyde (MDA) and nitric oxide (NO), while increasing superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX). In conclusion, spinach produced significant antioxidant effects, increased body weight, reduced the number of oocysts and parasite stages in the caecum, and restored the number of goblet cells relative to those of an uninfected control. Furthermore, spinach extract inhibits the sporulation percentage of E. tenella oocysts. The ethanolic extract of S. oleracea (whole plant) contained high concentrations of fatty acids, palmitic acid, Phytol, betulin, and ursolic aldehyde, all of which are known to regulate the antioxidant pathway and modulate inflammatory processes and may be the main reason for its anticoccidial activity.

The effect of stabilized culture of Lactobacillus rhamnosus GCC-3 on gut and liver health, and anti-viral immunity of zebrafish

Probiotics are promising antibiotics alternatives to improve growth and disease resistance of cultured fish. Our study aimed to investigate the effect of dietary stabilized culture of Lactobacillus rhamnosus GCC-3 on growth performance, gut and liver health and anti-viral ability of zebrafish (Danio rerio). Zebrafish (0.161 ± 0.001 g) were fed control and the experimental diet containing 1% GCC-3 culture (1 × 107 CFU/g diet) for four weeks. Growth performance and gut and liver health parameters were monitored after four weeks feeding. The gut microbiota was analyzed by 16S rRNA gene sequencing. In another experiment, zebrafish (0.212 ± 0.001 g) were fed with basal or GCC-3 diets and challenged by spring viremia of carp virus (SVCV) at the end of feeding. The antiviral immune response was evaluated at 2nd and 4th days post SVCV infection and survival rate was calculated 14 days after challenge. The results showed that adding 1% GCC-3 significantly improved growth performance of zebrafish (P < 0.05). The intestinal expression of hypoxia-inducible factor Hif-1α, tight junction protein ZO-1α and ZO-1β was significantly up-regulated in 1% GCC-3 group compared with control (P < 0.05). Besides, 1% GCC-3 decreased the content of MDA and increased total antioxidant capacity in the intestine, and the relative expression of SOD, GST and Gpxa was improved. The abundance of Proteobacteria was reduced while Firmicutes was enriched in the intestinal microbiota of 1% GCC-3 group compared with control (P < 0.05). Zebrafish fed 1% GCC-3 showed higher survival rate after SVCV challenge. Accordingly, the expression of antiviral genes in the spleen was increased at 2nd and 4th days post infection. In conclusion, our results indicate that dietary 1% GCC-3 supplementation can improve gut and liver health as well as antiviral immunity of zebrafish.

Characterization and Comparative Genomic Analysis of a Deep-Sea Bacillus Phage Reveal a Novel Genus.

As the most abundant biological entities, viruses are the major players in marine ecosystems. However, our knowledge on virus diversity and virus-host interactions in the deep sea remains very limited. In this study, vB_BteM-A9Y, a novel bacteriophage infecting Bacillus tequilensis, was isolated from deep-sea sediments in the South China Sea. vB_BteM-A9Y has a hexametric head and a long, complex contractile tail, which are typical features of myophages. vB_BteM-A9Y initiated host lysis at 60 min post infection with a burst size of 75 PFU/cell. The phage genome comprises 38,634 base pairs and encodes 54 predicted open reading frames (ORFs), of which 27 ORFs can be functionally annotated by homology analysis. Interestingly, abundant ORFs involved in DNA damage repair were identified in the phage genome, suggesting that vB_BteM-A9Y encodes multiple pathways for DNA damage repair, which may help to maintain the stability of the host/phage genome. A BLASTn search of the whole genome sequence of vB_BteM-A9Y against the GenBank revealed no existing homolog. Consistently, a phylogenomic tree and proteome-based phylogenetic tree analysis showed that vB_BteM-A9Y formed a unique branch. Further comparative analysis of genomic nucleotide similarity and ORF homology of vB_BteM-A9Y with its mostly related phages showed that the intergenomic similarity between vB_BteM-A9Y and these phages was 0-33.2%. Collectively, based on the comprehensive morphological, phylogenetic, and comparative genomic analysis, we propose that vB_BteM-A9Y belongs to a novel genus under Caudoviricetes. Therefore, our study will increase our knowledge on deep-sea virus diversity and virus-host interactions, as well as expanding our knowledge on phage taxonomy.

A CRISPR-based rapid DNA repositioning strategy and the early intranuclear life of HSV-1.

The relative positions of viral DNA genomes to the host intranuclear environment play critical roles in determining virus fate. Recent advances in the application of chromosome conformation capture-based sequencing analysis (3 C technologies) have revealed valuable aspects of the spatiotemporal interplay of viral genomes with host chromosomes. However, to elucidate the causal relationship between the subnuclear localization of viral genomes and the pathogenic outcome of an infection, manipulative tools are needed. Rapid repositioning of viral DNAs to specific subnuclear compartments amid infection is a powerful approach to synchronize and interrogate this dynamically changing process in space and time. Herein, we report an inducible CRISPR-based two-component platform that relocates extrachromosomal DNA pieces (5 kb to 170 kb) to the nuclear periphery in minutes (CRISPR-nuPin). Based on this strategy, investigations of herpes simplex virus 1 (HSV-1), a prototypical member of the human herpesvirus family, revealed unprecedently reported insights into the early intranuclear life of the pathogen: (I) Viral genomes tethered to the nuclear periphery upon entry, compared with those freely infecting the nucleus, were wrapped around histones with increased suppressive modifications and subjected to stronger transcriptional silencing and prominent growth inhibition. (II) Relocating HSV-1 genomes at 1 hr post infection significantly promoted the transcription of viral genes, termed an 'Escaping' effect. (III) Early accumulation of ICP0 was a sufficient but not necessary condition for 'Escaping'. (IV) Subnuclear localization was only critical during early infection. Importantly, the CRISPR-nuPin tactic, in principle, is applicable to many other DNA viruses.

Pictorial Review of Post COVID-19 Acute Invasive Fungal Sinusitis: Clinical Radiologists' Perspective.

Acute invasive fungal sinusitis (AIFS) is a rapidly progressive infection predominantly seen in immunocompromised patients. There is an increasing incidence of AIFS post COVID-19 infection. Yet, there is sparse literature regarding spectrum of cross-sectional imaging findings (CT and MRI) in these patients, which is prudent for appropriate timely surgical intervention. This study aims to highlight spectrum of imaging findings in patients with post-COVID-19 AIFS integrating the clinical details from presentation to follow-up. We retrospectively reviewed the demographics, clinical details and radiological imaging of 31 histopathological proven cases of post COVID-19 AIFS. MR and CT images of these patients were retrieved from the PACS and analysed. 90-day follow-up of these patients was obtained. Statistical analysis was performed using descriptive statistics. Cross-sectional imaging showed nasal cavity involvement in all patients, bilateral in (15, 48%) cases; predominant involvement of maxillary (31, 100%) followed by ethmoid sinuses (29, 93.5%) was seen. MR showed patchy/complete loss of normal mucosal enhancement in the turbinates and sinuses as predominant imaging finding. Maxillary sinus walls erosions were seen in 28 cases (90%). Rhino-orbital and rhino-orbito-cerebral mycosis was seen in 24 (77%) and 14 (45%) respectively. Optic nerve-sheath complex was involved in 15 (48%) cases. Cerebral involvement was seen in form of meningeal enhancement, cerebritis, ischemic changes, cavernous sinus and intracranial arterial thrombosis and aneurysms. Comprehensive knowledge of imaging features of AIFS and recognition of extent of their spread allows radiologists to play pivotal role in alerting the clinician for appropriate therapy to avoid protracted and fatal outcome.

Development of short hairpin RNA expression vectors targeting the internal ribosomal entry site of the classical swine fever virus genomic RNA

BackgroundClassical swine fever (CSF) is a fatal contagious disease affecting pigs caused by classical swine fever virus (CSFV). The disease can be transmitted by pigs and wild boars, and it is difficult to prevent and control. To obtain necessary information to establish the CSFV resistant animals in a future study, we designed lentiviral vector-delivered short hairpin RNAs (shRNAs) targeting the conserved domain III of the internal ribosomal entry site (IRES) of the CSFV genomic RNA.ResultsFirst, we confirmed the effects of siRNAs on CSFV-IRES activity. We observed significant inhibition of CSFV-IRES activity by si42 (domain IIIa), si107 (domain IIIc), and si198 (domain IIIf) in SK-L cells and si56 (domain IIIb), si142 (domain IIId1) and si198 in HEK293 cells without affecting the amount of luciferase RNA. Next, we constructed lentiviral vectors expressing shRNA based on siRNA sequences. Treatment with shRNA-expressing lentivirus was examined at 7 and 14 days post infection in SK-L cells and HEK293 cells, and CSFV-IRES was significantly suppressed at 14 days (sh42) post infection in HEK293 cells without significant cytotoxicity. Next, we examined the silencing effect of siRNA on CSFV replicon RNA and observed a significant effect by si198 after 2 days of treatment and by shRNA-expressing lentivirus (sh56, sh142, and sh198) infection after 14 days of treatment. Treatment of sh198-expressing lentivirus significantly suppressed CSFV infection at 3 days after infection.ConclusionThe IRES targeting sh198 expressing lentivirus vector can be a candidate tool for CSFV infection control.

Adenovirus type D and type E infection in broiler chickens: the effect on CD4 and CD8 T cell response, cytokines expression and their immunopathology

ABSTRACT 1. A total of 150-day-old chicks were divided into three groups of 50 birds (G1-G3); G1 and G2 were orally inoculated at 1-day old with 0.5 ml of 107 TCID50/ml FAdV-D serotype 2 (MT386509.1) and FAdV-E serotype 8a (MW847902), respectively, and G3 was blank control group. 2. Cell-mediated immune response was evaluated by detection of CD4, CD8 T lymphocytes and the mRNA expression of IL6 and IL8 in the chicken spleen using q-PCR. Additionally, immunopathology was performed at 3, 5 and 7 day post infection (dpi) and weekly until the end of the experiment. 3. Results revealed that transcription of inflammatory cytokines (IL6, IL8) was up regulated in the spleen of FAdV type D and type E infected chickens at various time points relative to the control group. A marked decrease in the number of CD4 and CD8 T lymphocytes at 5 and 7 dpi in G1 of chickens infected with FAdV type D. Whereas, in chickens infected with FAdV type E, the CD4 and CD8 T lymphocytes were markedly decreased at 7 dpi. 4. In contrast, there were no significant differences in humoral immune responses against NDV vaccine in (G1 and G2) at different intervals post-vaccination compared to the control group. The histopathology of the bursa, thymus, and spleen in the infected groups showed lymphocytolysis with severe reticular cells hyperplasia and lymphoid depletion. 5. In conclusion, fowl adenovirus types D and E have an immunosuppressive effect in broilers which may be considered one of the main causes of the continuous co-infections with other viruses reported in the field during the last 10 years.

Evaluation of zinc oxide and copper oxide nanoparticles as potential alternatives to antibiotics for managing fowl typhoid in broilers.

Antimicrobial resistance poses challenges to humans and animals, especially to the poultry sector in control of fowl typhoid with antibiotics, leading to increased mortality and food insecurity. Therefore, it is essential to develop more effective medications as alternatives to antibiotics. Currently, zinc oxide and copper oxide nanoparticles are of such significant interest due to their antibacterial properties. This study aimed to evaluate antimicrobial activity of zinc oxide and copper oxide nanoparticles against fowl typhoid in broilers. Ninety broiler chicks were raised under suitable management conditions. On day 10 of age, chicks were divided into six groups: control negative, control positive, T1, T2, T3, and T4. On day 19 of age, chicks in all groups except control negative were infected with Salmonella gallinarum (0.2 mL, 108 CFU/mL). After appearance of clinical signs, the treatments (Florfenicol; 50 mg/L drinking water [T1], and zinc oxide + copper oxide nanoparticles; 25 + 10 mg/kg/d [T2], 37.5 + 15 mg/kg/d [T3], and 50 + 20 mg/kg/d [T4]) were administered to chicks. Chicks were sacrificed on 26th and 30th day of age, and samples of blood and tissue were obtained. Hematological analysis with gross and histopathological examination of spleen, thymus and bursa of Fabricius was performed. Results revealed that there was no visible congestion in spleen and thymus of T3 and T4 at 11th day post infection. Antibody level against new castle's disease and lymphoproliferative response showed no significant difference in all groups. However, phagocytic response in nanoparticles treated groups exhibited a notable (p < 0.01) distinction compared to control positive. Notably, T3 demonstrated the highest level of phagocytic activity. Hematological parameters, including lymphocytes, heterophils, eosinophils, and heterophils/lymphocytes ratio in groups T2, T3, and T4, indicated significant (p < 0.01) difference compared to control positive. However, lymphocytes, heterophils, and heterophils/lymphocytes ratio in groups T2, T3, and T4 showed no significant difference when compared to T1. Nanoparticle treated groups showed decreased (p < 0.01) congestion of spleen and thymus as compared to control positive. Overall, zinc oxide and copper oxide nanoparticles have potential to serve as an alternative to florfenicol in treatment of fowl typhoid.

Inflammatory reaction and immune response of half-smooth tongue sole (Cynoglossus semilaevis) after infection with Vibrio anguillarum

Frequently occurred bacterial diseases have seriously affected the aquaculture industry of half-smooth tongue sole (Cynoglossus semilaevis). Notably, vibriosis, with Vibrio anguillarum as one of the causative pathogens, is the most severe bacterial disease with severe inflammatory response of the host, leading to high mortality rates. In the present study, we explored the relationship between bacterial concentrations and host mortality, inflammatory reaction, and immune response in half-smooth tongue sole after infection with V. anguillarum at different concentrations (Treatment 1, 6.4 × 105 CFU/mL; Treatment 2, 6.4 × 106 CFU/mL). The mortality of Treatment 2 (77.5%) was significantly higher than that of Treatment 1 (10%), corresponding with bacterial concentrations. Although the number of deaths varies, intensive deaths were observed within 24 h post infection (hpi) in both bacterial concentration groups. Histopathological analyses revealed that fish tissues were most severely damaged at 24 or 48 hpi, and Treatment 2 was more severe than Treatment 1. A qRT-PCR-based detection method with virulence factor gene empA was established to quantify the bacterial loads in various tissues, and the bacterial loads were the highest at 24 hpi in Treatment 2, and at 48 hpi in Treatment 1. Additionally, the expression levels of complement genes (C5a, C3, C5, and C6), inflammatory factors (IL-1β, TNF-α, and IL-10), and other immune-related genes (jak2, NF-κB1, stat3, and tlr3) were increased in various tissues after infection in both treatment groups, with most genes being most expressed at 24 or 48 hpi, and expression levels of inflammatory factors in Treatment 2 were higher than those in Treatment 1. Moreover, the expression of C5a was positively correlated with that of proinflammatory cytokines in both bacterial concentration groups. According to the results of this study, 24–48 hpi was a key node for early vibriosis detection and intervention. Compared with the low mortality of Treatment 1, the mass death of fish in Treatment 2 was suggested to be caused by uncontrolled excessive inflammatory reaction induced by the overactivation of complement system, especially C5a. We believe these results could provide theoretical basis for prevention, evaluation, and treatment of vibrio disease in tongue sole aquaculture, and lay a solid foundation for future functional analyses.

Evaluating anti-viral effect of Ivermectin on porcine epidemic diarrhea virus and analyzing the related genes and signaling pathway by RNA-seq in vitro

Porcine epidemic diarrhea virus (PEDV) has catastrophic impacts on the global pig industry. However, there remains no effective drugs for PEDV infection. Ivermectin is an FDA-approved anthelmintic drug used to treat worm infections. In this study, we reported the broad-spectrum antiviral activity of Ivermectin in vitro. Ivermectin can inhibit PEDV infections of different genotypes. Avermectin derivatives can also inhibit PEDV infections. A time of addition assay showed that Ivermectin exhibited potent anti-PEDV activity when added simultaneously with or post virus infection. Furthermore, Ivermectin significantly inhibited the late stage of viral infection by affecting viral release. RNA sequencing indicates Ivermectin induces cell cycle arrest, which may be related to its ability to inhibit viral release. Interestingly, when combined with Niclosamide, Ivermectin demonstrated an enhanced anti-PEDV effect. These findings highlight Ivermectin as a novel antiviral agent with potential for the development of drugs against PEDV infection.

Methyltransferase-like 3 suppresses red spotted grouper nervous necrosis virus and viral hemorrhagic septicemia virus infection by enhancing type I interferon responses in sea perch.

Methylation at the N6 position of adenosine (m6A) is the most abundant internal mRNA modification in eukaryotes, tightly associating with regulation of viral life circles and immune responses. Here, a methyltransferase-like 3 homolog gene from sea perch (Lateolabrax japonicus), designated LjMETTL3, was cloned and characterized, and its negative role in fish virus pathogenesis was uncovered. The cDNA of LjMETTL3 encoded a 601-amino acid protein with a MT-A70 domain, which shared the closest genetic relationship with Echeneis naucrates METTL3. Spatial expression analysis revealed that LjMETTL3 was more abundant in the immune tissues of sea perch post red spotted grouper nervous necrosis virus (RGNNV) or viral hemorrhagic septicemia virus (VHSV) infection. LjMETTL3 expression was significantly upregulated at 12 and 24 h post RGNNV and VHSV infection in vitro. In addition, ectopic expression of LjMETTL3 inhibited RGNNV and VHSV infection in LJB cells at 12 and 24 h post infection, whereas knockdown of LjMETTL3 led to opposite effects. Furthermore, we found that LjMETTL3 may participate in boosting the type I interferon responses by interacting with TANK-binding kinase. Taken together, these results disclosed the antiviral role of fish METTL3 against RGNNV and VHSV and provided evidence for understanding the potential mechanisms of fish METTL3 in antiviral innate immunity.

Abstract P436: Increased West Nile Virus Infection And Pathology In The Spontaneously Hypertensive Rat

Introduction: West Nile virus (WNV), a mosquito-borne flavivirus, causes grave neuroinvasive disease and death, and has been reported in over 55,000 people in the US since 1999. Patients &gt;55 years and those with hypertension are most at risk for WNV-associated mortality. Understanding how hypertension correlates with WNV morbidity and mortality may reduce the burden of this disease. Investigation of WNV in hypertensive people is limited to basic epidemiology. Without an animal model, understanding the causal relationship between hypertension and WNV-associated pathology remains elusive. Here, we tested a novel hypothesis that hypertension will promote WNV infection and pathology in a rodent model of age-dependent chronic hypertension, the spontaneously hypertensive rat (SHR). Methods: Male adult SHR and normotensive WKY rats were implanted with radiotelemetry transmitters for measurements of blood pressure and heart rate. Following baseline measurements, SHR and WKY rats (N=9/gp) were injected with WNV (1 X 10 4 PFU/ml, i.p.). Vehicle injection (PBS, i.p.) was performed in additional WKY and SHR (N=2-3/gp). Each rat was observed for clinical abnormalities, and body weight, blood pressure, and heart rate were measured daily. Rats were sacrificed on days 9 and 10 post-infection (PI). At endpoint, several tissues were processed for measurement of viral load by RT-qPCR and histopathology. Bone marrow and circulating immune cells (CD4 + , CD8 + , CD68 + ) were quantified at endpoint by flow cytometry. Results: One SHR rat developed neurological symptoms and was euthanized on day 4 PI. Viral load was significantly increased in the SHR kidney and spleen (P&lt;0.001) in SHR compared to WKY rats. In addition, WNV-infection elevated heart rate in the SHR but not the WKY (P&lt;0.0001). SHR also showed elevated circulating CD68 + monocytes (P&lt;0.05), and a significant increase (P&lt;0.05) in renal pathology was observed in the SHR following WNV infection. Conclusions: We demonstrate, for the first time, a positive interaction between high blood pressure and severity of WNV infection in a rodent model of chronic hypertension. Further studies are needed to elucidate the mechanisms of this interaction for new therapeutic targets of WNV disease.

Distribution of carp edema virus in organs of infected juvenile common carp.

The disease caused by carp edema virus (CEV) manifests with lethargy as a primary sign; this observation in koi in Japan gained the disease the name koi sleepy disease (KSD). In the years following the discovery of the virus in Japan, KSD cases have been noted in the UK in koi and common carp. Conducting research in order to expand knowledge of the processes of distribution of CEV in infected fish organs will be helpful for eradication and diagnostic purposes. Carp edema virus-affected fish with clinical signs of KSD were experimentally cohabited with common carp fry (30 fish). Three fish were euthanised by bath in a 0.5 g L-1 tricaine solution at one week intervals (7, 14, 21 and 28 days post cohabitation). Tissue samples from the brain, gills, spleen, kidney, intestines and skin were collected, and the total DNA was extracted and tested by real-time PCR. By the seventh day post infection, CEV DNA was most often found in the skin, gills and brain and less frequently in the kidney and intestines. In many of the common carp fry, CEV DNA could typically be found in several organs of each individual fish, although it was only found in one sample of spleen tissue. In this experimental study the pathogenesis of the CEV infection process was shown, the high infectivity of CEV was confirmed and the best organs were determined for sampling in CEV-infection experimentation. The real-time PCR method used in our cohabitation experiments was shown to be useful at the clinical and asymptomatic stage of virus infection.

Efficacy of Raphamin against Pneumococcal Infection: a Preclinical Study.

The aim of the study was to evaluate the activity of Raphamin in a model of non-lethal pneumococcal infection caused by Streptococcus pneumoniae 3 in BALB/c mice. The drug or placebo was administered intragastrically 3 days prior to infection, 2 h before and 2 h post infection, and then for 3 full days, alone or in combination with antibiotic (amoxicil-lin/clavulanic acid). Raphamin monotherapy significantly decreased bacterial load in the lungs in comparison with placebo (p<0.05) which was comparable to the effect in antibiotic alone or combined with Raphamin. Raphamin prevented reproduction of Streptococcus pneumoniae in the lower respiratory tract and its combination with the antibiotic was safe and did not reduce the efficacy of amoxicillin/clavulanic acid.

Effect of COVID vaccination on monthly migraine days: a longitudinal cohort study.

This longitudinal cohort study aimed to investigate changes in migraine-related outcomes following COVID-19 infection and vaccination. We identified 547 clinically diagnosed migraine patients from the Leiden Headache Center who kept a headache E-diary during the COVID-19 pandemic (February 2020 to August 2022). We sent a questionnaire to register their COVID-19 infection and/or vaccination dates. After applying inclusion criteria, n = 59 participants could be included in the infection analysis and n = 147 could be included in the vaccination analysis. Primary outcome was the change in monthly migraine days (MMD) between 1 month prior and 1 month post COVID-19 infection or vaccination. Secondary outcome variables were change in monthly headache days (MHD) and monthly acute medication days (MAMD). Vaccination against COVID-19 was associated with an increase in MMD (1.06; 95% confidence interval [CI] = 0.57-1.55; p < 0.001), MHD (1.52; 95% CI = 0.91-2.14; p < 0.001) and MAMD (0.72; 95% CI = 0.33-1.12; p < 0.001) in the first month post-vaccination. COVID-19 infection solely increased the number of MAMD (1.11; 95% CI = 0.10-1.62; p < 0.027), but no statistically significant differences in MMD or MHD were observed. Our findings imply that vaccination against COVID-19 is associated with an increase in migraine, indicating a possible role of inflammatory mediators in migraine pathophysiology.