A comprehensive review on lipid nanocarrier systems for cancer treatment: fabrication, future prospects and clinical trials.

Over the last few decades, cancer has been considered a clinical challenge, being among the leading causes of mortality all over the world. Although many treatment approaches have been developed for cancer, chemotherapy is still the most utilized in the clinical setting. However, the available chemotherapeutics-based treatments have several caveats including their lack of specificity, adverse effects as well as cancer relapse and metastasis which mainly explains the low survival rate of patients. Lipid nanoparticles (LNPs) have been utilized as promising nanocarrier systems for chemotherapeutics to overcome the challenges of the currently applied therapeutic strategies for cancer treatment. Loading chemotherapeutic agent(s) into LNPs improves drug delivery at different aspects including specific targeting of tumours, and enhancing the bioavailability of drugs at the tumour site through selective release of their payload, thus reducing their undesired side effects on healthy cells. This review article delineates an overview of the clinical challenges in many cancer treatments as well as depicts the role of LNPs in achieving optimal therapeutic outcomes. Moreover, the review contains a comprehensive description of the many LNPs categories used as nanocarriers in cancer treatment to date, as well as the potential of LNPs for future applications in other areas of medicine and research.

Potential anticancer effect of free and nanoformulated Deferasirox for breast cancer treatment: in-vitro and in-vivo evaluation.

BACKGROUND: Breast cancer (BC) stands as the second-leading cause of mortality among women worldwide. Many chemotherapeutic treatments for BC come with significant adverse effects. Additionally, BC is recognized as one of the most resistant forms of malignancy to treatment. Consequently, there exists a critical need for innovative therapeutic agents that are both highly effective and exhibit reduced toxicity and side effects for patients. Deferasirox (DFX), an iron-chelating drug approved by the FDA for oral use, emerges as a promising contender in the fight against BC proliferation. DFX, primarily administered orally, is utilized to address chronic iron excess resulting from blood transfusions, and it is the inaugural treatment for chronic iron overload syndrome. However, DFX encounters limitations due to its poor water solubility. AIM: This study aimed at incorporating DFX into lipid nanocapsules (DFX-LNCs) followed by investigating the anticancer effect of the DFX nanoform as compared to free DFX in-vitro and on an orthotopic BC mouse model in-vivo. METHODS: The DFX-LNCs was prepared and imaged using TEM and also characterized in terms of particle size (PS), zeta potential (ZP), and polydispersity index (PDI) using DLS. Moreover, drug release, cytotoxicity, and anticancer effect were assessed in-vitro, and in-vivo. RESULTS: The results revealed that DFX-LNCs are more cytotoxic than free DFX with IC50 of 4.417 µg/ml and 16.114 µg/ml, respectively, while the plain LNCs didn't show any cytotoxic effect on the 4T1 cell line (IC50 = 122.797 µg/ml). Besides, the apoptotic effect of DFX-LNCs was more pronounced than that of free DFX, as evidenced by Annexin V/PI staining, increased BAX expression, and decreased expression of BcL-2. Moreover, DFX-LNCs showed a superior antitumor effect in-vivo with potent antioxidant and anti-proliferative effects. CONCLUSION: The newly developed DFX nanoform demonstrated a high potential as a promising therapeutic agent for BC treatment.

Early pathogenesis in rabbit hemorrhagic disease virus 2.

To detail early tissue distribution and innate immune response to rabbit hemorrhagic disease virus 2 (RHDV2), 13 rabbits were orally (Oryctolagus cuniculus) inoculated with liver homogenate made from a feral rabbit that succumbed to RHDV2 during the 2020 outbreak in Oregon, USA. Rabbits were monitored regularly, with euthanasia and collection of tissues and swabs, at 12, 24, 36, 48, 96, and 144 h post inoculation. Livers from these rabbits were positive by RT-rtPCR for presence of the virus. Using RNAscope for viral and replicative intermediates, rabbits had detectable viral genomic RNA at each time point, initially within the gastrointestinal tract, then in the liver by 36 h post inoculation. Also using RNAscope, there were increasing amounts of mRNA coding for TNF-α, IL-6, and IL-1ß within the liver and spleen through 48 h post inoculation. The results of this study aided our understanding of the local innate immune response to RHDV2, as well as aspects of pathogenesis.

Impacts of deficiency in vitamin D derivatives on disease severity in adult bronchial asthma patients.

Vitamin D affects innate and adaptive immunity processes that impact treatment, severity, and morbidity of acute asthma episodes. Several vitamin D forms may help modulate immunity, including vitamin D2 (D2), vitamin D3 (D3), 25-hydroxyvitamin D3 (25(OH)D3), and 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3). This study assessed serum levels of vitamin D derivatives in bronchial asthma patients and their correlation with disease markers. One hundred thirteen subjects, divided into two groups, were enrolled. The first group included 73 asthmatic patients (57 males and 16 females), and the second included 40 healthy adults (31 males and 9 females) as a control group. All subjects were evaluated with a careful history and clinical examination, a chest X-ray with a posteroanterior view, routine laboratory examination, spirometry, and asthma control tests (ACT). Vitamin D serum levels were assessed using ultra-performance liquid chromatography (UPLC) with tandem mass spectrometry. Disease markers were assessed and correlated with serum levels of vitamin D forms. Markers included forced expiratory volume in 1 s (FEV1), forced vital capacity (FVC), FEV1/FVC%, peak expiratory flow (PEF), forced expiratory flow25-75% (FEF25-75%), eosinophilic blood count, and total immunoglobulin E (IgE). Asthmatic patients had significantly lower serum levels of vitamin D than healthy controls (p ≤ 0.001). Further, serum vitamin D levels decreased significantly in uncontrolled asthmatic patients than partially controlled and controlled patients. Correlations for 25(OH)D3 and 1,25-(OH) 2D3 were stronger than for D2 and D3. There were negative correlations for eosinophilic blood count, total IgE, and ACT. Serum levels of all vitamin D forms were reduced in asthmatic patients with moderate to strong correlations with disease severity. Vitamin D deficiency or even insufficiency may thus play a role in disease severity.

Deferasirox: A comprehensive drug profile.

Deferasirox is an iron-chelating drug developed by Novartis company for treatment of diseases accompanied by chronic iron overload; such as ß-thalassemia or sickle cell diseases. Owing to its advantages such as high affinity, specificity and wide therapeutic window, it is considered as first line treatment. The current chapter describes the physicochemical characteristics, mode of action, pharmacokinetics, therapeutic applications and synthetic methods for deferasirox. Moreover, it includes Fourier transform infrared spectrometry (FTIR) and nuclear magnetic resonance spectroscopy (NMR) analysis for its functional groups. In addition, the selected analytical methods are summarized to aid the analysts in their routine analysis of deferasirox.

A Case of Renal Artery Thrombosis With Concurrent Adrenal Hemorrhage in Polycythemia Vera.

Polycythemia vera (PV) is a rare myeloproliferative neoplasm characterized by the clonal proliferation of hematopoietic stem cells, leading to an elevated red blood cell mass. This hyperproliferative state increases blood viscosity and predisposes patients to thrombotic events, which are a significant cause of morbidity and mortality in PV. The diagnosis of PV is typically confirmed through elevated hemoglobin or hematocrit levels, low serum erythropoietin, and the presence of the Janus kinase 2 (JAK2) mutation. Common complications include venous and arterial thromboses, hemorrhage, and transformation to myelofibrosis or acute leukemia. A 68-year-old female with a history of PV and chronic kidney disease (CKD) presented with uremic symptoms in the form of malaise and nausea. Laboratory investigations indicated acute kidney injury (AKI) and hyperkalemia. Imaging evaluation of renal US Doppler revealed renal artery thrombosis and an incidental adrenal hemorrhage. The patient was managed with intravenous heparin and did not receive thrombolytics or thrombectomy. Her renal function did not improve, necessitating the initiation of hemodialysis (HD) during hospitalization. Over the course of the next few weeks, her renal parameters improved and she managed to be discharged from dialysis. The primary goal of this study was to highlight a rare presentation of renal artery thrombosis secondary to polycythemia vera (PV) and discuss the complexities involved in managing the underlying disease and its thrombotic complication, particularly in the presence of concomitant bleeding. Effective management of PV-related thrombosis requires a delicate balance between anticoagulation to prevent further thrombotic events while carefully addressing the risk of hemorrhage.

Coding-complete genome sequences of rabbit hemorrhagic disease virus type 2 detected in 2023 in Washington State indicate a divergent incursion into the United States.

Three rabbit hemorrhagic disease virus type 2 (RHDV2) coding-complete genome sequences were obtained from domestic and wild rabbits in Washington State in June and July 2023. These three RHDV2 sequences are <82% identical to previous RHDV2 sequences in North America and likely indicate a discrete incursion.

Recent progress on polySarcosine as an alternative to PEGylation: Synthesis and biomedical applications.

Biotherapeutic PEGylation to prolong action of medications has gained popularity over the last decades. Various hydrophilic natural polymers have been developed to tackle the drawbacks of PEGylation, such as its accelerated blood clearance and non-biodegradability. Polypeptoides, such as polysarcosine (pSar), have been explored as hydrophilic substitutes for PEG. pSar has PEG-like physicochemical characteristics such as water solubility and no reported cytotoxicity and immunogenicity. This review discusses pSar derivatives, synthesis, characterization approaches, biomedical applications, in addition to the challenges and future perspectives of pSar based biomaterials as an alternative to PEG.

Plasmodium vivax Malaria Complicated With Acute Respiratory Distress Syndrome: A Case Report.

Plasmodium vivax (P. vivax) malaria was long considered to have low mortality. Severe malaria was classically associated with Plasmodium falciparum (P. falciparum). However, there is growing evidence that severe and complicated forms of vivax malaria may be more widespread than previously assumed. We report a case of a 32-year-old female patient with acute respiratory distress syndrome (ARDS) as a complication of P. vivax malaria who has recovered completely following starting intravenous antimalarial medications and noninvasive ventilation (NIV).

Polyvinyl alcohol-chitosan-polyethylene glycol-glycerol incorporated with Peganum harmala loaded in lipid nanocapsules as an elastic nanocomposite surgical sealant to control bleeding.

Uncontrolled hemorrhage remains a critical threat in trauma and surgery. This study developed a novel hemostatic composite by encapsulating Peganum harmala L. seed extract (PH) with known hemostatic properties into lipid nanocapsules (PH-LNCs) and then embedding them within a polyvinyl alcohol-chitosan-polyethylene glycol-glycerol (PVA-CS-PEG-G) matrix. The composite was physically crosslinked via the dual processes of freezing-thawing and thermal crosslinking and exhibited robust mechanical properties reaching 0.434 ± 0.014 MPa and elasticity of 40.685 % ± 4.04. It also demonstrated excellent biocompatibility, surface morphology, physical stability, and ex-vivo skin deposition/permeation were assessed. The characterization of PH-LNCs revealed optimal PH-LNC formation and successful integration into the composite with particle size, zeta potential, and PDI were approximately 45.45 ± 24 nm, -16.3 ± 1.4 mV, and 0.374 ± 0.1, respectively. In vitro studies highlighted enhanced blood clotting and platelet adhesion, while in vivo experiments confirmed superior hemostatic efficacy in a mouse tail amputation model. The composite's soft texture, conformability, and mechanical strength make it a promising candidate for effective traumatic wound management.

Folic/lactobionic acid dual-targeted polymeric nanocapsules for potential treatment of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a malignant tumor that affects many patients diagnosed with hepatic cell inflammation and liver cirrhosis. Targeted polymeric nanocapsules could facilitate the internalization and accumulation of anticancer drugs. Dual-targeted folic acid/lactobionic acid-poly lactic co-glycolic acid nanocapsules (NCs) were prepared and loaded with pterostilbene (PTN) and characterized for their physicochemical properties, as well as in vitro and in vivo anticancer activity. NCs displayed a size of 222 nm, zeta potential of - 16.5 mV, and sustained release for 48 h. The IC50 of PTN NCs (5.87 ± 0.8 µg/mL) was 20 times lower than unencapsulated PTN (121.26 ± 9.42 µg/mL) on HepG2 liver cancer cells owing to the enhanced cellular uptake of the former, as delineated by flow cytometry. In vivo study on HCC-induced animals delineated the superiority of the dual-targeted NCs over the unencapsulated PTN, which significantly reduced the liver markers ALT, AST, and ALP, as well as the tumor-related markers AFP and Bcl2, and elevated the anti-apoptotic marker caspase 3. Furthermore, the NCs significantly reduced the oxidative stress and exhibited almost comparable histological features to the normal group. Therefore, it can be concluded that the dual-ligated folic acid/lactobionic acid nanocapsules can be considered a promising potential treatment option for hepatocellular carcinoma.

Student Performance and Perceptions of Flipped Classrooms and Small Group Discussions as Teaching Tools in Practical Anatomy.

Introduction A quasi-experimental study was conducted to assess students' attitudes toward the flipped practical classroom and evaluate its effectiveness in teaching practical anatomy. Methods Two survey questionnaires were developed: the first assessed students' attitudes toward flipped practical classrooms, while the second focused on the obstacles and difficulties students might encounter in this learning environment. Results We found that students rated flipped learning significantly higher compared to other teaching methods, particularly in terms of the quality of learning materials, enhancement of learning skills, engagement and understanding of anatomy topics, and problem-solving abilities. The highest mean examination grades were observed for the pretest flipped modality, followed by the pretest small group discussion (mean scores: 82.72 vs. 54.46, F = 43.2, P = 0.004). Conclusions Students hold positive attitudes toward flipped classrooms and small group discussions compared to traditional classes.

Antibody response of endangered riparian brush rabbits to vaccination against rabbit hemorrhagic disease virus 2.

Rabbit hemorrhagic disease virus 2 (RHDV2; Caliciviridae, Lagovirus europaeus), the cause of a highly transmissible and fatal lagomorph disease, has spread rapidly through the western United States and Mexico, resulting in substantial mortality in domestic and wild rabbits. The disease was first detected in California in May 2020, prompting an interagency/zoo/academia/nonprofit team to implement emergency conservation actions to protect endangered riparian brush rabbits (Sylvilagus bachmani riparius) from RHDV2. Prior to vaccinating wild rabbits, we conducted a vaccine safety trial by giving a single SC dose of Filavac VHD K C+V (Filavie) vaccine to 19 adult wild riparian brush rabbits captured and temporarily held in captivity. Rabbits were monitored for adverse effects, and serum was collected before vaccination, and at 7-10, 14-20, and 60 d post-vaccination. Sera were tested using an ELISA to determine antibody response and timing of seroconversion. Reverse-transcription quantitative real-time PCR (RT-qPCR) was performed on rectal swabs to evaluate infection status. No adverse effects from the vaccine were observed. Before vaccination, 18 of 19 rabbits were seronegative, and RHDV2 was not detected by RT-qPCR on any rectal swabs. After vaccination, all rabbits developed an antibody response, with titers of 1:10-1:160. Seroconversion generally occurred at 7-10 d. The duration of antibody response was ≥60 d in 12 of 13 rabbits. Sixteen animals were released and 4 were recaptured several months later, offering a glimpse into longer duration immune response. Our study has informed vaccination strategies for this species and serves as a model for protecting other vulnerable lagomorphs against RHDV2.

Tsunami deposits in Tunisia contemporaneous of the large 365 CE Crete earthquake and Mediterranean Sea catastrophic event.

New field investigations along the East Tunisian coastline reveal sedimentary deposits and damaged localities that may account for a catastrophic event during the late Holocene. North of Sfax city, ~ 3.4 m high cliff coastal marine and alluvial terraces show 20 to 50-cm-thick chaotic layer with sandy coarse gravels mixed with reworked pebbles, broken shells of gastropods and molluscs, organic matter and Roman pottery. The chaotic layer truncates sandy-silty paleosol, covers Roman settlements and is overlain by fire remains, a thin sandy-silty aeolian unit and ~ 1-m-thick alluvial deposits. Charcoal samples collected at 10 cm below and 4 cm above the catastrophic deposits provide radiocarbon dating that brackets the catastrophic unit between 286 and 370 CE. Other historical investigations on the Roman sites of Neapolis (Nabeul), Hadrumete (Sousse), Thyna (Sfax), Meninx in Girba (Djerba), Wadi Ennouili (Gulf of Gabes), and Sabratha (in Libya) evidenced major damage and abandonment of sites in the fourth century (16, 41, 42, 43, 44). The new identification of catastrophic deposits, offshore-onshore correlations with turbidites and modelling of tsunami waves suggest the relationship with the 21 July 365 tsunamigenic earthquake (Mw ~ 8) of west Crete (Greece) and call for a better estimate of tsunami risk on the Mediterranean coastlines.

Role of virtual bronchoscopy in the evaluation of bronchial lesions.

BACKGROUND: Virtual bronchoscopy (VB) is a type of 3-dimensional reconstruction in which the observation point is placed within the airway to produce an endoscopiclike view. AIM: To evaluate the diagnostic role of VB in the diagnosis of tracheobronchial lesions, as compared to fiberoptic bronchoscopy (FOB). SUBJECTS: Fifty patients with tracheobronchial lesions were enrolled (30 patients with bronchogenic carcinomas and 20 patients with tracheobronchial inflammatory lesions). METHODS: The patients were examined using VB and FOB. Virtual bronchoscopic studies were calculated and reconstructed from cross-sectional images obtained from spiral computed tomographic examination of the chest. RESULTS: Virtual bronchoscopy provided an excellent overview of the trachea, main stem, and lobar bronchi up to the fourth order. The data obtained by VB and FOB (signs of tumor infiltration including endobronchial mass, stenosis, obstruction, and external indentations) were comparable. However, FOB had the advantage of giving direct cues to color, vascularity, and motility. It also detected early tumor infiltration by picking up subtle mucosal changes. Alternatively, VB was superior in bypassing any obstruction and therefore provided an excellent view distal to the obstructive lesions or stenotic segments. Virtual bronchoscopy also defined the optimum pathway for passing instruments into lesions beyond the field of view. CONCLUSIONS: Here, we compared the diagnostic capacities of both VB and FOB. The interventional and therapeutic ramifications of our findings await further investigations.

Impact of photodynamic therapy as an adjunct to non-surgical periodontal treatment on clinical and biochemical parameters among patients having mild rheumatoid arthritis with periodontitis.

PURPOSE: To evaluate the efficacy of photodynamic therapy (PDT) as an adjunct to non-surgical periodontal therapy on the clinical periodontal and biochemical parameters among patients with rheumatoid arthritis (RA) having periodontitis. METHODS: A total of 50 RA patients with periodontitis were included. The subjects were equally divided into two groups: Group A - scaling and root planning (SRP) + PDT; Group B - SRP only, respectively. Plaque score (PS), bleeding on probing (BOP), and pocket depth (PD) were estimated. The biochemical parameters included the assessment of interleukin 6 (IL-6), tumor necrosis factor-alpha (TNF-α) and rheumatoid factors (RFs). RESULTS: Plaque scores and BOP significantly reduced in both the groups at both 6 and 12 weeks with significant difference between both the groups at 6 weeks follow up (p<0.05). On inter-group comparison, there was a statistically significant reduction seen for BOP in Group A at 12 weeks (p<0.001). PD significantly reduced in both the groups at both time points; however, significant reduction was noted for Group A compared to Group B (p<0.01). IL-6 and TNF-α significantly reduced in both the groups at 6 and 12 weeks follow up. However, the proinflammatory cytokine levels significantly reduced in group A as compared to group B at both 6 and 12 weeks (p<0.05). GCF levels of RF did not show any change in either of the groups at either time point or between the groups (p>0.05). CONCLUSION: PDT significantly reduced the proinflammatory burden in terms of periodontal attachment level and bleeding on probing within the periodontal inflammatory pockets in patients having RA.

Switching indication of PEGylated lipid nanocapsules-loaded with rolapitant and deferasirox against breast cancer: Enhanced in-vitro and in-vivo cytotoxicity.

Breast cancer (BC) is considered the leading cause of mortality and morbidity among adult women worldwide, and it is associated with many genetic or hormonal factors. Despite the advanced therapeutic and theranostic strategies for BC treatment, cancer metastasis and relapse are often observed among patients which lead to therapeutic failure. Accordingly, among the repositioned medication against BC proliferation is neurokinin receptor antagonists and iron chelating agents especially rolapitant HCl (RP) and deferasirox (DFO), respectively. However, RP and DFO are classified as class II with low aqueous solubility. Both drugs were nanoformulated into PEGylated lipid nanocapsules (LNCs) for enhancing their aqueous solubility and augmenting their efficacy. RP-LNCs, DFO-LNCs and their combinations were evaluated according to particle size (PS), zeta potential, polydispersity index (PDI) and surface morphology. Importantly, the antitumor effect of these novel molecules and their nanoforms was evaluated against the suppression of Ehrlich Ascites tumor model using female mice. Results revealed that RP-LNCs, DFO-LNCs and RP/DFO-LNCs exerted PS from 45.23 ± 3.54 to 60.1 ± 3.32 nm with PDI around 0.20 which indicates homogenous particles distribution. Also, RP-LNCs, DFO-LNCs and RP/DFO-LNCs displayed surface charges of +16.6 ± 6.9, -13.3 ± 5.82 and - 20.2 ± 5.40 mV, respectively. The obtained LNCs conferred a high potent cytotoxic effect against MCF7 cancer cells as compared to parent drugs, with IC50 of 10.86 ± 0.89, 3.34 ± 0.99 and 2.24 ± 0.97 µg/mL for RP-LNCs, DFO-LNCs and RP/DFO-LNCs, respectively. The in-vivo pharmacodynamics effect of the developed nano-formulations showed superior antitumor effect for the individual drugs rather than their combinations as compared to the control group. The current study confirmed the potential of RP and DFO nanoforms as promising therapeutic agents for BC treatment.

New repurposed rolapitant in nanovesicular systems for lung cancer treatment: Development, in-vitro assessment and in-vivo biodistribution study.

Lung cancer is characterized by poor prognosis, and is considered a serious disease that causes a significant mortality. The available conventional chemotherapeutic agents suffer from several limitations; hence, new drug molecules are constantly being sought. In the current study, lipid nanovesicles (LNVs) were selected as a colloidal vehicle for encapsulation of the FDA-approved drug; rolapitant (RP), which is used particularly for the treatment of nausea and vomiting, but is repurposed for the treatment of lung cancer in the current work. RP was loaded into various LNVs (liposomes, ethosomes and transethosomes) using the thin film hydration method, and the LNVs were evaluated for particle size, zeta potential, entrapment efficiency (EE%), storage stability and surface morphology. Besides, the in-vitro drug release, in-vitro cytotoxicity on A549 lung cancer cells, nebulization performance using next generation impactor (NGI), and the in-vivo biodistribution behavior were evaluated. The selected ethosomal and transethosomal vesicles displayed a particle size less than 400 nm, a positive charge, and EE% exceeding 90% for RP, with a sustained release pattern over 15 days. The in-vivo biodistribution results proved the high lung deposition potential of RP-LNVs with a considerable safety. Besides, the developed RP-LNVs were able to reach the metastatic organs of lung cancer, hence they were proven promising as a possible treatment modality for lung cancer.

Comparative susceptibility of eastern cottontails and New Zealand white rabbits to classical rabbit haemorrhagic disease virus (RHDV) and RHDV2.

Rabbit haemorrhagic disease virus (RHDV) is associated with high morbidity and mortality in the European rabbit (Oryctolagus cuniculus). In 2010, a genetically distinct RHDV named RHDV2 emerged in Europe and spread to many other regions, including North America in 2016. Prior to this study it was unknown if eastern cottontails (ECT(s); Sylvilagus floridanus), one of the most common wild lagomorphs in the United States, were susceptible to RHDV2. In this study, 10 wild-caught ECTs and 10 New Zealand white rabbits (NZWR(s); O. cuniculus) were each inoculated orally with either RHDV (RHDVa/GI.1a; n = 5 per species) or RHDV2 (a recombinant GI.1bP-GI.2; n = 5 per species) and monitored for the development of disease. Three of the five ECTs that were infected with RHDV2 developed disease consistent with RHD and died at 4 and 6 days post-inoculation (DPI). The RHDV major capsid protein/antigen (VP60) was detected in the livers of three ECTs infected with RHDV2, but none was detected in the ECTs infected with RHDV. Additionally, RHD viral RNA was detected in the liver, spleen, intestine and blood of ECTs infected with RHDV2, but not in the ECTs infected with RHDV. RHD viral RNA was detected in urine, oral swabs and rectal swabs in at least two of five ECTs infected with RHDV2. One ECT inoculated with RHDV2 seroconverted and developed a high antibody titre by the end of the experimental period (21 DPI). ECTs inoculated with the classic RHDV did not seroconvert. In comparison, NZWRs inoculated with RHDV2 exhibited high mortality (five of five) at 2 DPI and four of five NZWRs inoculated with RHDV either died or were euthanized at 2 DPI indicating both of these viruses were highly pathogenic to this species. This experiment indicates that ECTs are susceptible to RHDV2 and can shed viral RNA, thereby suggesting this species could be involved in the epidemiology of this virus.