Antiviral effects of bovine lactoferricin-lactoferrampin delivered via recombinant lactobacillus on senecavirus a and foot-and-mouth disease virus in mice

Аннотация

Senecavirus A (SVA) and Foot-and-Mouth Disease Virus (FMDV) are significant viral pathogens that cause major economic losses in livestock, particularly in pigs and cattle. This study investigates the inhibitory effects of bovine lactoferricin-lactoferrampin (bLFcin-LFamp) peptides on SVA and FMDV, delivered through recombinant Lactobacillus oral treatment in mice. The peptides' antiviral activity was evaluated in vitro and in vivo, with Lactobacillus engineered to express bLFcin-LFamp as a delivery vehicle. The results demonstrated that the recombinant Lactobacillus significantly reduced viral replication and clinical symptoms in mice infected with both SVA and FMDV. This study suggests that bLFcin-LFamp peptides, when delivered via oral Lactobacillus treatment, represent a promising therapeutic strategy for controlling viral infections in livestock. Keywords: Bovine lactoferricin, lactoferrampin, Senecavirus A, Foot-and-Mouth Disease Virus, recombinant Lactobacillus, antiviral treatment, mice, oral delivery, viral inhibition, peptides. Introduction: Senecavirus A (SVA) and Foot-and-Mouth Disease Virus (FMDV) are highly contagious viral pathogens that pose significant threats to the global livestock industry. SVA is an emerging picornavirus that affects swine, causing vesicular lesions and economic losses in pig farming. FMDV, a member of the The American Journal of Veterinary Sciences and Wildlife Discovery 05 https://www.theamericanjournals.com/index.php/tajvswd The American Journal of Veterinary Sciences and Wildlife Discovery Picornaviridae family, affects cloven-hoofed animals, To assess the antiviral properties of bLFcin-LFamp, in including cattle, sheep, and pigs, leading to severe vitro assays were performed using Vero cells infected production losses and trade restrictions. Both viruses with SVA and FMDV. Cells were pretreated with various are capable of causing extensive morbidity and concentrations of recombinant Lactobacillus expressing mortality in affected populations. bLFcin-LFamp for 1 hour before viral inoculation. After 48 hours of incubation, viral titers were measured using Currently, there are limited antiviral treatments the TCID50 (tissue culture infective dose) method to available for these viral infections, with the focus evaluate the antiviral efficacy of the peptides. primarily on vaccination and quarantine measures. The development of novel antiviral strategies is essential 4. Animal Model and Treatment Protocol for enhancing disease management, especially in the Six-week-old female BALB/c mice were used for in vivo face of emerging viral strains that may not be studies. Mice were divided into four groups: (1) effectively controlled by existing vaccines. uninfected control, (2) SVA-infected, (3) FMDV-infected, Bovine lactoferricin and lactoferrampin (bLFcin- and (4) treatment groups (SVA- or FMDV-infected mice LFamp) are antimicrobial peptides derived from bovine treated with recombinant Lactobacillus expressing lactoferrin, a protein with broad-spectrum bLFcin-LFamp). Mice in the treatment groups were antimicrobial and antiviral properties. Previous studies orally administered 1 × 10^9 CFU of recombinant have demonstrated that these peptides exhibit activity Lactobacillus daily for 7 days before viral infection and against various pathogens, including bacteria, fungi, continuing throughout the course of the infection. and viruses. This study aims to explore the potential Infection was performed by intraperitoneal injection antiviral effects of bLFcin-LFamp peptides on SVA and with 1 × 10^5 TCID50 of either SVA or FMDV. Clinical FMDV, with a novel approach of delivering these signs, such as weight loss, fever, and the presence of peptides through recombinant Lactobacillus bacteria vesicular lesions, were monitored daily. At day 7 post- for oral administration in a mouse model. infection, animals were euthanized, and organs were This research investigates the following: harvested for viral load quantification via quantitative PCR (qPCR) and histopathological examination. 1. The antiviral activity of bLFcin-LFamp against SVA and FMDV in vitro. 5. Viral Load Quantification and Statistical Analysis 2. The use of recombinant Lactobacillus to Viral loads in serum and tissue samples were quantified deliver bLFcin-LFamp in vivo. by qPCR using specific primers for SVA and FMDV. The results were expressed as log10 TCID50 equivalents per 3. The efficacy of oral treatment with milliliter (mL). Statistical analysis was performed using recombinant Lactobacillus expressing bLFcin-LFamp in one-way ANOVA, followed by Tukey’s post-hoc test to reducing viral replication and clinical symptoms in compare viral load and clinical outcomes between mice. treatment groups. P-values < 0.05 were considered METHODS statistically significant. 1. SVA and FMDV Strains