Feline calicivirus (FCV) typically causes severe respiratory infections and oral ulcers in cats, posing a significant threat to feline health. However, the high variability of FCV presents serious challenges to vaccine development and drug treatment. Currently available vaccines may not provide complete protection; thus, the need to screen for novel broad-spectrum vaccine strains to enhance cross-protection against multiple genotypes is urgent. This study successfully established animal models with a 100
Poxviruses (family Poxviridae) infect a wide range of human and animal hosts, causing diseases of substantial concern in both public health and veterinary medicine. Although smallpox was eradicated by global vaccination, the multicountry spread of mpox since 2022 and the continuing burden of capripox and parapox infections underscore the need for sustained poxvirus research and preparedness. Poxviruses have a complex virion architecture with two infectious forms, the intracellular mature virion (IMV) and extracellular enveloped virion (EEV), and large DNA genomes encoding more than 200 proteins, creating an antigenic landscape that complicates cross-genus comparison and application-oriented target prioritization. To define the biological basis for rational poxvirus antigen selection, we provide a cross-genus comparative synthesis of antigenic proteins in Orthopoxvirus, Capripoxvirus, and Parapoxvirus, integrating biological function, sequence conservation, antibody recognition, and published immunogenicity data. We delineate major antigen classes associated with IMV, EEV, and core structures and evaluate how their conserved and variable features influence immunodominance, neutralization, cross-protection, and serological discrimination. By relating these antigen-level features to translational application, this review establishes an antigen-centered framework that connects fundamental biology with practical relevance and provides a comparative basis for vaccine candidate selection, serological differentiation, and antibody-based intervention.
Tembusu virus (TMUV) is an emerging flavivirus that causes encephalitis and a decrease in egg production in poultry across East and Southeast Asia. Here, we isolated and characterized a chicken-origin TMUV strain, CHN-WH, from a laying hen farm in China. Whole-genome sequencing and phylogenetic analysis based on the complete open reading frame revealed that CHN-WH belongs to cluster 3.2 and is most closely related to the mosquito-derived strain TMUV-YN2020. Comparative sequence analysis and structural modeling revealed several cluster 3.2-specific signature residues in the E protein, as well as a strain-specific M198L substitution predicted to be located in a surface-exposed region. CHN-WH replicated efficiently in BHK-21 cells, chicken embryo fibroblasts (CEFs), and duck embryo fibroblasts (DEFs). In vivo, CHN-WH caused systemic viral dissemination and neurological lesions in chicks and induced ovarian lesions accompanied by reduced egg production in laying hens. These findings indicate that CHN-WH is a pathogenic chicken-origin cluster 3.2 TMUV strain with distinct genomic and phenotypic characteristics. This study expands the current knowledge of the molecular features and pathogenic potential of the chicken-origin cluster 3.2 TMUV and provides useful information for epidemiological surveillance and vaccine development.
Schmallenberg virus (SBV), an emerging Culicoides‑borne arbovirus, is responsible for febrile illness and reduced milk production in adult ruminants and can induce congenital malformations in fetuses, representing a substantial concern for livestock health worldwide. Consequently, rapid, field‑adaptable diagnostic approaches are urgently needed. In this study, we developed a visual nucleic acid detection assay for SBV that combines reverse transcription recombinase‑aided amplification (RT‑RAA) with the CRISPR/Cas12a system, targeting conserved regions of the SBV S gene. Following systematic optimization of the reaction conditions, the assay was completed within 50 min, with a sensitivity of 8.6 copies/μL for the SBV-S plasmid and 8.6 × 101 copies/μL for the RNA transcripts. The assay exhibited high specificity, with no cross‑reactivity observed against a panel of relevant pathogens, including Seoul orthohantavirus (SEOV), infectious bovine rhinotracheitis virus (IBRV), Rift Valley fever virus (RVFV), Crimean‑Congo hemorrhagic fever virus (CCHFV), and bovine viral diarrhea virus (BVDV). When evaluated using simulated clinical samples, the sensitivity of the method was superior to that of conventional real-time fluorescent reverse transcription‒polymerase chain reaction (RT‑qPCR). Importantly, the entire reaction is performed in a single closed-tube format, significantly reducing the risk of cross-contamination and false-positive results. In summary, this method demonstrates favorable analytical performance for the detection of SBV-S plasmid-spiked nasal swab samples and holds promise for further clinical validation, although its diagnostic utility in authentic clinical specimens remains to be confirmed in future investigations.
Abstract H9N2 avian influenza virus (AIV) remains a global threat to poultry health and has zoonotic potential. Antigenic drift in the hemagglutinin (HA) protein complicates vaccine efficacy and diagnostic accuracy, highlighting the need for precise epitope characterization. In this study, the HA protein of H9N2 AIV was expressed in a eukaryotic system, and two monoclonal antibodies (mAbs), 9C12 and 9F4, were generated. Both mAbs specifically bound HA, as shown by ELISA, Western blot, and immunofluorescence, but lacked hemagglutination inhibition activity. Epitope mapping revealed two minimal linear epitopes: 123FSSSRSYQ130 within the vestigial esterase domain and 201NLYTRTDTT209 within the receptor-binding domain. Alanine scanning revealed key residues required for antibody binding, whereas structural modeling confirmed that both epitopes are surface exposed. Sequence analysis demonstrated strong conservation across H9N2 strains, with the 9F4 epitope showing near-complete invariance, whereas both epitopes exhibited low conservation among other influenza A virus subtypes. These findings define two novel, nonneutralizing epitopes on H9N2 HA that expand the antigenic map and represent promising targets for subtype-specific diagnostic assays.
Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne zoonotic pathogen of increasing global concern that is maintained through intricate interactions among Hyalomma ticks, livestock, wildlife, and humans. Among wildlife species, wild ungulates exhibit substantial seroprevalence across endemic regions and may contribute to the maintenance and dispersal of infected ticks at the wildlife–livestock interface. However, most available evidence from wildlife remains serological and should not be interpreted as definitive proof of reservoir competence or sustained vertebrate-mediated viral maintenance. In this review, I compiled the available evidence of natural and experimental CCHFV infection in wild animal species, with a particular emphasis on ungulates. Existing surveillance efforts remain largely focused on human infections and domestic livestock, whereas the ecology of wildlife-associated transmission remains relatively underexplored. Improved surveillance of wild ungulates and tick populations, combined with molecular detection and geospatial analysis of host–vector interactions, could strengthen the understanding of viral circulation in endemic ecosystems and support more accurate risk prediction. A One Health approach that integrates wildlife, livestock, vector, and human surveillance is therefore essential for improving ecological understanding and supporting evidence-based mitigation strategies for CCHFV.
Abstract Salmonella enterica serovar Enteritidis (SE) not only causes systemic acute infections in chickens but can also be transmitted to humans through contaminated poultry and egg products, posing a serious threat to public health safety. In this study, we constructed a multi-epitope subunit vaccine against SE and investigated its immunostimulatory capacity and protective efficacy in mice and chickens. Ten B-cell epitopes and three T-cell epitopes were selected from two immunogenic proteins, FliC and SipD, and linked to construct a multi-epitope vaccine designated FLPD. In silico analysis revealed that FLPD exhibited favorable immunogenicity, stability, and immune-inducing potential. To validate its protective efficacy in vivo, immunization and challenge experiments were performed in mice and chickens. In mice, FLPD induced high IgG levels, with the 50 μg dose group showing particularly robust responses. After triple immunization, FLPD conferred 65% and 45% protection against lethal challenge with SE and Salmonella Typhimurium (ST), respectively. In chickens, triple immunization with FLPD and a commercial live attenuated vaccine achieved 70% and 80% protection against lethal SE challenge, respectively. Furthermore, FLPD-induced polyclonal antibodies showed cross-reactive effects against eight common Salmonella serovars in vitro. These findings suggest that FLPD is a promising vaccine candidate for combating SE infections.
Abstract Feline infectious peritonitis (FIP) is a fatal disease caused by feline coronavirus (FCoV), a member of the Coronaviridae family that comprises two genotypes, FCoV-I and FCoV-II. From the ascitic fluid of a British shorthair cat diagnosed with FIP, we isolated and identified an FCoV strain designated AH2023. Western blotting, RT‒PCR, and transmission electron microscopy were employed for characterization. According to phylogenetic analyses of the full viral genome and the spike, nucleocapsid, and 3C-like protease genes, strain AH2023 consistently fell within the clade of previously reported Chinese FCoV-II strains and exhibited greater evolutionary proximity to transmissible gastroenteritis virus (TGEV) and canine coronavirus (CCoV) than to FCoV-I did. Recombination analysis using SimPlot further revealed strain AH2023 to be a recombinant virus. Strain AH2023 is genetically distinct, carrying four amino acid substitutions in the spike protein. Substitutions at positions 387, 577, and 829 are shared with strains SH2021 and DQ2023, whereas the substitution at position 1375 is shared with strain DF2. Pathogenicity tests in 12‑week‑old cats confirmed the virulence of AH2023. All the cats died between 12 and 18 days post-challenge (dpc) and presented severe intestinal, hepatic, and renal lesions. The virus exhibited broad tissue tropism, with the highest viral load (10 4 copies/mg) found in the intestine. Fecal shedding began at 7 dpc and reached 10 4 copies by 10 dpc. These results advance our understanding of FCoV evolution and pathogenesis, especially for type II strains in China, and aid in the development of effective FIP prevention strategies.
Brucella species are responsible for the zoonotic disease brucellosis and utilize the type IV secretion system (T4SS) to introduce effector proteins into host cells, thereby modulating immune responses and sustaining intracellular infection. Notably, BspJ was identified as the first Brucella nucleomodulin. Although our previous studies demonstrated its influence on host inflammatory responses and bacterial colonization, the precise molecular mechanism remains elusive. In this study, we report novel findings that BspJ directly targets the host transcription factor Zfp740 promoter region through three critical amino acid residues (His66, Arg87, and Arg96), leading to the upregulation of Zfp740 expression. This upregulation subsequently blocks IκBα degradation and inhibits P65 phosphorylation and nuclear translocation, thereby obstructing the activation of the NF-κB signaling pathway and ultimately diminishing the production of proinflammatory cytokines. Moreover, both the BspJ deletion strain and strains with mutations in key amino acids significantly enhanced the inflammatory response and compromised the intracellular survival capacity of Brucella. In conclusion, our study systematically elucidates a novel regulatory axis, BspJ-Zfp740-P65/NF-κB, and reveals a new mechanism by which Brucella directly interferes with host transcriptional programs through a nucleus-targeting effector protein to suppress inflammation and establish persistent infection. These findings provide a significant theoretical foundation for understanding bacteria-mediated immune evasion and chronic infection and introduce new research avenues for the development of host-directed anti-Brucella strategies.
Rabies virus (RABV) is a neurotropic virus that infects the central nervous systems of humans and animals, causing fatal neurological symptoms and ultimately death. Vaccination is the only effective strategy for preventing rabies. However, once clinical symptoms appear, there is no effective treatment. Therefore, exploring effective therapeutic approaches, particularly for postexposure prophylaxis, is imperative. This study investigated the inhibitory effect of siRNA delivered by recombinant adeno-associated virus serotype 2 (rAAV2) against RABV. Specific siRNAs targeting the mRNA of the G and L genes of the RABV were designed. rAAV-G997 and rAAV-L5055 were constructed with siRNA expression cassettes. Pretreatment of neuroblastoma (NA) cells with either rAAV-G997 or rAAV-L5055 before RABV infection resulted in a considerable reduction in both the viral titer and the mRNA levels of the G and L genes. Similarly, pretreatment of baby hamster kidney (BHK-21) cells with rAAV-G997 or rAAV-L5055 prior to RABV infection decreased the expression of viral structural proteins. In a mouse model, compared with the negative control (rAAV-NC), the intracranial administration of rAAV-G997 or rAAV-L5055 conferred significant protection. When administered before challenge with the RABV CVS-11 strain, both rAAV-G997 and rAAV-L5055 significantly reduced the viral load in brain tissue, and achieved a survival rate of 90
Abstract Viral host tropism defines the molecular boundaries of infection and governs the adaptive processes by which pathogens cross species barriers. This review synthesizes the mechanistic and evolutionary determinants that enable viral spillover, with a focus on receptor recognition, proteolytic activation, immune evasion, and genomic plasticity as core drivers of host range expansion. By integrating structural virology, comparative genomics, and molecular immunology, we examine how adaptive mutations, recombination, and host‒virus coevolution shape molecular compatibility and facilitate interspecies transmission. We further analyzed how environmental and anthropogenic pressures, including habitat disruption, climate change, and wildlife‒livestock‒human interfaces, modulate viral evolution and increase exposure opportunities. Central to this synthesis is the proposed sequential molecular gatekeeping model of viral spillover, which outlines the stepwise molecular and ecological barriers that viruses must overcome to achieve successful cross-species emergence. This review highlights how advances in metagenomic surveillance and artificial intelligence (AI)-driven prediction of receptor binding affinity can identify high-risk viral lineages before they cause outbreaks. Framed within a One Health perspective, this work bridges molecular mechanisms, ecological dynamics, and translational preparedness, offering a conceptual framework to support hypothesis generation and risk-informed surveillance. The SMGM integrates previously fragmented molecular determinants into a structured, stage-based conceptual model rather than introducing entirely new mechanistic components.
Abstract Toxoplasma gondii ( T. gondii) is a globally distributed protozoan parasite that can infect a wide range of warm-blooded animals, including humans. Nonhuman primates (NHPs) are particularly valuable for studying toxoplasmosis because of their physiological and immunological similarity to humans. In this study, we report the isolation and characterization of two T. gondii strains from captive NHPs that died suddenly in a zoo in Guangdong Province, China. The animals, a red-backed bearded saki ( Chiropotes chiropotes ), a New World monkey, and a mandrill ( Mandrillus sphinx ), an Old World monkey, were diagnosed with acute fatal toxoplasmosis on the basis of histopathology, immunohistochemistry, serology (MAT), and PCR. Parasite isolation was performed via a mouse bioassay, and the strains were genotyped using multilocus PCR–RFLP. The isolates were identified as ToxoDB #15 and ToxoDB #169, atypical genotypes that have been reported in Africa, Europe, and the Americas. Pathogenicity assessment in Swiss mice revealed that both TgMonkeyGd2 (ToxoDB #15) and TgMonkeyGd3 (ToxoDB #169) exhibited intermediate virulence, causing 100% and 80% mortality at 10 4 tachyzoites, respectively, with lower doses allowing host survival and seroconversion. These findings challenge the traditional view that Old World monkeys (exemplified by the mandrill) are relatively resistant to toxoplasmosis and suggest a possible role of the wildlife trade in the transcontinental spread of exotic T. gondii genotypes, although this hypothesis requires further investigation with a larger number of cases. Enhanced surveillance and biosecurity measures are urgently needed in captive animal facilities to prevent outbreaks and protect both animal and public health.
Abstract Porcine rotavirus (PoRV) induces severe gastroenteritis in both suckling and weaned piglets, resulting in substantial economic losses to the global swine industry. In this study, a group A PoRV (PoRVA) strain, designated RShanD1, was isolated from the intestinal contents of a piglet suffering from severe diarrhea. Whole-genome sequencing analysis revealed that the genotype of RShanD1 is G5-P[7]-I5-C1-M1-R1-A1-N1-T1-E1-H1. Phylogenetic analysis indicated that RShanD1 is a porcine-bovine reassortant rotavirus. By cotransfecting 11 plasmids encoding viral structural and nonstructural proteins, along with a plasmid expressing the African swine fever virus (ASFV) capping enzyme NP868R, into BHK-T7 cells (baby-hamster kidney cells stably expressing T7 RNA polymerase), we established a complete reverse genetics system for RShanD1. Compared with those of the parental strain wtRShanD1, the plaque sizes and growth kinetics of the rescued virus, rRShanD1, were comparable. We also assessed the pathogenicity of rRShanD1 and wtRShanD1 using a suckling mouse infection model, and the results demonstrated that both parental wtRShanD1 and rescued rRShanD1 tended to exhibit similar changes in average daily weight, viral shedding, and intestinal histopathology. Overall, we isolated and identified a porcine–bovine reassortant rotavirus and successfully established a reverse genetics system, which not only facilitates the study of PoRV biology but also paves the way for advancing the development of PoRV vaccines.
Abstract Mastitis remains the most economically significant disease in dairy production, impacting milk yield and quality from the mammary gland and impairing overall animal welfare. Conventional treatment strategies rely heavily on antibiotics, raising concerns about antimicrobial resistance, withdrawal periods, and consumer demand for residue-free milk. Photobiomodulation (PBM), a noninvasive therapy utilizing low-level laser light, has emerged as a promising adjunct for udder health management. This review synthesizes current evidence on PBM’s physiological mechanisms, therapeutic benefits, and practical implications for dairy herd health. PBM operates through the activation of mitochondrial chromophores, primarily cytochrome c oxidase, triggering enhanced ATP synthesis, nitric oxide release, and redox signaling. These processes modulate inflammatory pathways, including NF-κB and MAPK signaling, and regulate cytokine production, thereby promoting tissue repair without broadly suppressing immune function. In cows, PBM has been shown to reduce oxidative stress, inflammatory mediators, and the somatic cell count (SCC), contributing to improved milk quality and accelerated recovery. Studies report up to a 60% reduction in subclinical mastitis cases and an enhanced recovery of 31.2% when PBM is combined with antibiotics, underscoring the role of PBM in antimicrobial stewardship. Beyond therapeutic outcomes, PBM offers welfare advantages by alleviating pain and minimizing stress during treatment. Its noninvasive nature reduces handling-related anxiety compared with conventional interventions. Emerging evidence also suggests potential benefits for milk composition (increase in de novo fatty acids) and metabolic stability, although further research is needed to confirm these effects in vivo. Despite its promise, PBM faces limitations, including variability in treatment parameters, restricted tissue penetration, and logistical challenges in large herds. Standardization of dosing protocols and cost-effectiveness analyses remain critical for widespread adoption. Thus, PBM represents a viable complementary strategy for mastitis management, aligning with industry goals to reduce antibiotic dependence while enhancing animal welfare and milk quality. Continued research into optimized applications, long-term efficacy, and economic feasibility will determine its integration into sustainable dairy health programs.
In this study, the hepatoprotective potential of Salvia splendens (S. splendens) leaf extract against atorvastatin-induced liver toxicity in Wistar albino rats was evaluated. Twenty-four rats were divided into four groups: a negative control group, a positive control group receiving atorvastatin (20 mg/kg/day), a standard treatment group receiving atorvastatin with silymarin (200 mg/kg/day), and an herbal treatment group receiving atorvastatin with S. splendens extract (200 mg/kg/day). Treatments were administered for 14 d. At the end of the experiment, blood and liver samples were collected for biochemical, hematological, and histopathological analyses. Atorvastatin administration caused a significant reduction in body weight gain and a marked increase in liver and renal function marker levels, along with increased oxidative stress. Compared with treatment with silymarin, treatment with S. splendens extract significantly improved body weight gain, restored the antioxidant status, and reduced the increase in liver enzymes. Histopathological findings supported the biochemical results, as hepatic damage was reduced in the treated groups. These findings suggest that S. splendens possesses significant hepatoprotective activity and may serve as a potential natural therapeutic agent against drug-induced liver injury.
Anthelmintic resistance in gastrointestinal nematodes poses a severe threat to global livestock production and sustainability. While the genomic adaptations of parasites have been extensively characterized, the role of the parasite-associated microbiome in modulating drug susceptibility remains largely unexplored. In this study, we investigated the symbiotic microbiota of ivermectin-resistant and ivermectin-susceptible strains of the blood-feeding parasitic nematode Haemonchus contortus and assessed its potential association with ivermectin resistance. Comparative microbiome analysis revealed Stenotrophomonas maltophilia as a potential resistance biomarker, with a 3.4-fold higher relative abundance in resistant strains. Furthermore, functional prediction analysis revealed that pathways related to genetic information processing and cell motility were significantly enriched in the microbiota of resistant strains. Larval mortality assays demonstrated that gentamicin treatment significantly increased the susceptibility of the larvae to ivermectin. These findings suggest that symbiotic bacteria are correlated with ivermectin resistance in Haemonchus contortus, possibly in combination with genetic factors of the parasite.
Abstract Streptococcus suis (S. suis) is the principal etiological agent of swine streptococcosis and is an emerging zoonotic pathogen. In recent years, the prevalence of S. suis serotype 9 (SS9) has transitioned from primarily subclinical colonization to invasive disease, accompanied by a continuous increase in isolation rates across multiple countries. However, compared with that of serotype 2 (SS2), our understanding of SS9 remains markedly limited. In this study, we analyzed 7,913 S. suis isolates, among which SS9 was the second most prevalent serotype, accounting for 9.12% (722/7,913), followed by SS2 at 26.9% (2,132/7,913). We also isolated a highly virulent SS9 meningitis-associated strain and performed a comprehensive genomic analysis of 722 SS9 isolates. Population structure analysis revealed substantial genetic diversity within SS9, comprising 168 sequence types (STs) and 23 CCs, with ST16 representing the predominant lineage. The predominance of isolates from the brain and cerebrospinal fluid (37.89%), together with the lack of source-specific phylogenetic clustering, suggests a potential association of SS9 with central nervous system-related infection, particularly meningitis. Bioinformatics analysis revealed significant differences in the distribution of 17 virulence genes between brain-associated and nonbrain-associated isolates, indicating a potential association between these genes and meningitis. The most common antimicrobial resistance genes were tet(O) (54.29%) and erm(B) (62.05%), with 36.98% of the strains carrying both resistance to tetracycline and resistance to MLSB antibiotics. This study was designed to investigate whether the meningitis-associated pathogenicity of SS9 is related to the enrichment of specific virulence determinants and multidrug resistance-associated genomic features, thereby enhancing our understanding of the pathogenicity and resistance of SS9 and providing important data for improving disease control strategies and public health preparedness.
Canine mammary tumors are challenging to treat and share multiple molecular and pathological features with human breast cancer, particularly triple-negative breast cancer (TNBC). Demethylzeylasteral (T-96) exhibits significant antitumor activity; however, its mechanism of action against triple-negative breast cancer (TNBC) remains poorly understood. Moreover, pharmacokinetic information regarding intraperitoneal administration remains limited in BALB/c mice. This study aimed to investigate the therapeutic efficacy and underlying mechanisms of T-96 against TNBC and to characterize its pharmacokinetic profile. Transcriptomic analysis revealed that T-96 modulates pathways related to the cell cycle and apoptosis. Consistently, protein-level validation and MDM2 perturbation experiments support the functional involvement of the MDM2–p21 signaling module in T-96-associated phenotypes. At the molecular level, T-96 upregulated the expression of p21, BAX, and cleaved caspase 9 but downregulated the expression of MDM2 and Bcl-2. Pharmacokinetic analysis revealed that T-96 reached its peak plasma concentration (Tmax) at 2.0 h postadministration and had a half-life (t1/2) of 23.5 h. In a mouse breast tumor model, T-96 significantly inhibited TNBC growth, increased the number of necrotic lesions in the tumor center, and reduced liver and lung metastases. Functional validation using transient MDM2 overexpression and inhibitor cotreatment supported a functional contribution of MDM2 to several T-96-associated phenotypes. To our knowledge, this study provides the first evidence that T-96 treatment is associated with reduced MDM2 levels and increased p21 expression, accompanied by G1 phase accumulation and activation of mitochondrial apoptosis markers in TNBC models, collectively contributing to the suppression of TNBC cell growth. Here, MDM2 was identified as a key functional mediator rather than a direct molecular target of T-96. Our findings indicate that the MDM2-p21 signaling module may partially functionally mediate T-96-associated cell cycle arrest and apoptosis, providing preclinical, comparative oncology–relevant evidence to support further evaluation of T-96 in additional models, including those of spontaneously occurring canine mammary tumors.
Abstract Ticks are obligate hematophagous ectoparasites that play a critical role in wildlife health and disease transmission. While Amblyomma helvolum is a well-known specialist parasite of reptiles, particularly the Komodo dragon (Varanus komodoensis), its infestation of atypical hosts remains poorly documented. This study describes the first confirmed case in which A. helvolum infested a human in Indonesia and discusses its broader implications for wildlife health and conservation management. A tick sample was recovered from a staff member involved in wildlife monitoring at Loh Buaya, Rinca Island, within Komodo National Park. Morphological examination revealed that the sample was an adult male A. helvolum, which was further corroborated by molecular analysis of the 16S rRNA gene, which revealed 99–100% similarity to reference sequences. Concurrent field observations also revealed the presence of A. helvolum on V. komodoensis within the same locality, highlighting an active tick‒host interface. This documented host-switching event serves as a biological sentinel, reflecting intense ectoparasite pressure within the native reptile population and the potential for pathogen spillover. These findings underscore the necessity of integrating ectoparasite surveillance into routine wildlife health monitoring and biosecurity protocols to safeguard both the endangered Komodo dragon and the personnel dedicated to its conservation in high-interaction habitats.
