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Mexico is home to 14 species of lagomorphs, 6 of which are endemic. Studies on diseases affecting native lagomorphs are scarce, and in most cases, the impact on their populations remains largely unknown. Rabbit hemorrhagic disease virus (RHDV), especially the RHDV2 variant, causes a serious and extremely contagious disease, resulting in high mortality rates and major declines in wild lagomorph populations. The objectives of this study were to identify disease hotspots and critical biodiversity regions in Mexico through the combined use of disease information and lagomorph distribution maps and to determine the areas of greatest concern. In total, 19 states of Mexico recorded RHDV2 from April 2020 to August 2021, and 12 of them reported the wild species Sylvilagus audubonii, Lepus californicus, and unidentified Leporidae species. The distribution of RHDV2 in Mexico can be closely predicted from climatic variables. RHDV2 hotspots are located in the central-southern area of the Mexican Highlands and the Trans-Mexican Volcanic Belt, where the virus affects multiple species. This knowledge is essential for proposing specific actions to manage and preserve lagomorph populations at risk and address these issues as soon as possible. [ABSTRACT FROM AUTHOR]

A new form of the rabbit haemorrhagic disease virus, RHDV2, first observed in European rabbits, has spread widely among different species of hares in Europe, jackrabbits and cottontails in North America, and hares in southern Africa. However, only limited surveillance studies have been undertaken so far. It is suggested that methods developed for controlling the disease in farmed rabbits in Europe and studying the efficacy of RHDV as a biological control agent in Australia could facilitate epidemiological research on those recently affected lagomorph species. This would enable the assessment of the risk of RHDV2 to native lagomorphs, including endangered species, and the determination of the main host species of RHDV2. Because RHDV2 has not spread equally through all lagomorph species, epidemiological studies could give insights into factors important for determining host susceptibility. [ABSTRACT FROM AUTHOR]

Trans fatty acids are specific unsaturated fats found in processed foods that undergo hydrogenation, leading to hepatic disorders such as metabolic-associated fatty liver disease (MAFLD) and conditions like CVD and CKD. The effects of different food samples containing trans fatty acids (elaidic and oleic acid) on the liver, heart, and kidney through antioxidant enzyme activity were investigated in animal models. Liver function tests (ALT, ALP, AST, and LDH), heart biomarker levels (CPK, TC, HDL, LDL, and triglycerides), and kidney biomarker levels (serum creatinine, blood urea nitrogen, and serum uric acid) were examined in serum of rabbits and the histopathology of liver tissues. Results showed that these biomarkers were more elevated in the Mujahid Ghee group than in the normal control, oleic acid, and Kausar Ghee groups. The concentration of antioxidant markers such as peroxidase, glutathione, catalase, thiobarbituric acid reactive substances, and superoxide dismutase were lower in the Mujahid Ghee group. HPLC showed that Mujahid Ghee had the highest quantified value of elaidic acid among all selected samples. Overall, this study demonstrated that elaidic acid in its purest form aggravated MAFLD in rabbit livers and provoked CVK and CVD. [ABSTRACT FROM AUTHOR]

Simple Summary: Mammary carcinomas have been diagnosed with increasing frequency in pet rabbits. Prognostic biomarkers are limited, and the only available treatment is surgical excision. Additional treatment options are needed, e.g., for animals in which metastases to internal organs preclude complete tumor removal. Human breast cancer may express the immunosuppressive enzyme indoleamine 2,3-dioxygenase 1 (IDO1), that represents a prognostic biomarker and a possible therapeutic target. Since previous studies revealed similarities between human breast cancer and pet rabbit mammary carcinomas, this study investigated IDO1 immunostaining in 96 mammary carcinomas of 96 pet rabbits with an average age of 5.5 years. All rabbits with reported sex were female. Variable percentages of IDO1-positive tumor cells were detected in 90 (94%) carcinomas. Furthermore, IDO1 immunostaining was observed in the secretory epithelial cells of the adjacent non-neoplastic mammary tissue. This study provides further information on the molecular features of mammary carcinomas in rabbits. It also shows similarities in IDO1 expression between rabbit mammary carcinomas and human breast cancer. These findings can have a mutual benefit. They could lead the development of novel treatment options for rabbits with mammary carcinomas. In addition, they further support the value of rabbits with mammary carcinomas for breast cancer research. For mammary carcinomas in pet rabbits, prognostic biomarkers are poorly defined, and treatment is limited to surgical excision. Additional treatment options are needed for rabbit patients for which surgery is not a suitable option. In human breast cancer, the immunosuppressive enzyme indoleamine 2,3-dioxygenase 1 (IDO1) represents a prognostic biomarker and possible therapeutic target. This retrospective immunohistochemical study examined IDO1 in 96 pet rabbit mammary carcinomas with known mitotic count, hormone receptor status, and percentage of stromal tumor infiltrating lymphocytes (TILs). Tumors were obtained from 96 pet rabbits with an average of 5.5 years. All rabbits with reported sex (n = 88) were female or female-spayed. Of the carcinomas, 94% expressed IDO1, and 86% had sparse TILs consistent with cold tumors. Statistically significant correlations existed between a higher percentage of IDO1-positive tumor cells, lower mitotic counts, and increased estrogen receptor expression. The threshold for significance was IDO1 staining in >10% of tumor cells. These results lead to the assumption that IDO1 expression contributes to tumorigenesis and may represent a prognostic biomarker and possible therapeutic target also in pet rabbit mammary carcinomas. They also support the value of rabbits for breast cancer research. [ABSTRACT FROM AUTHOR]

Simple Summary: The study focuses on the shadow of prion protein (Sho) encoded by the shadow of prion protein gene (SPRN), which associates with PrP and promotes the progression of prion diseases. While genetic polymorphisms in SPRN are linked to susceptibility in several species, rabbit SPRN gene polymorphisms have not been extensively studied. Through amplicon sequencing, we identified novel single nucleotide polymorphisms (SNPs) in the rabbit SPRN gene and found strong linkage disequilibrium (LD) between them. However, strong LD was not observed between polymorphisms in the prion protein gene (PRNP) and SPRN genes in rabbits. Comparison of amino acid sequences revealed differences in SPRN between rabbits and other species susceptible or resistant to prion diseases. This study represents the first examination of genetic features of the rabbit SPRN gene. Prion disorders are fatal infectious diseases that are caused by a buildup of pathogenic prion protein (PrPSc) in susceptible mammals. According to new findings, the shadow of prion protein (Sho) encoded by the shadow of prion protein gene (SPRN) is associated with prion protein (PrP), promoting the progression of prion diseases. Although genetic polymorphisms in SPRN are associated with susceptibility to several prion diseases, genetic polymorphisms in the rabbit SPRN gene have not been investigated in depth. We discovered two novel single nucleotide polymorphisms (SNPs) in the leporine SPRN gene on chromosome 18 and found strong linkage disequilibrium (LD) between them. Additionally, strong LD was not found between the polymorphisms of PRNP and SPRN genes in rabbits. Furthermore, nonsynonymous SNPs that alter the amino acid sequences within the open reading frame (ORF) of SPRN have been observed in prion disease-susceptible animals, but this is the first report in rabbits. As far as we are aware, this study represents the first examination of the genetic features of the rabbit SPRN gene. [ABSTRACT FROM AUTHOR]

Simple Summary: In recent years, research has shown that animals' gut microbiota can regulate various physiological functions such as nutrition, metabolism, and immunity, and play an indispensable role in maintaining host gut health. At present, there are few reports on the establishment of rabbits' gut microbiota and its relationship with the host. To clarify the colonization of rabbits' gut microbiota, the changes in species during development, and the relationship between gut microbiota and rabbits' growth and development, this study determined the 16s rRNA of gut microbiota in rabbits of different age groups and preliminarily explored the changes in the types of gut microbiota during the colonization process. Six amplicon sequence variants (ASVs) that significantly affect body weight were obtained. The research results provide a firm theoretical basis for maintaining gut health and constructing new methods for rabbit breeding. This study used feces from 0-day-old (36 rabbits), 10-day-old (119 rabbits), and 60-day-old (119 rabbits) offspring rabbits and their corresponding female rabbits (36 rabbits) as experimental materials. Using 16s rRNA sequencing, the study analyzed the types and changes of gut microbiota in rabbits at different growth and development stages, as well as the correlation between gut microbiota composition and the weight of 60-day-old rabbits. All experimental rabbits were placed in the same rabbit shed. Juvenile rabbits were fed solid feed at 18 days of age and weaned at 35 days of age. In addition to identifying the dominant bacterial phyla of gut microbiota in rabbits at different age stages, it was found that the abundance of Clostridium tertium and Clostridium paraputrificum in all suckling rabbits (10-day-old) was significantly higher than that in rabbits fed with whole feed (60-day-old) (p < 0.05), while the abundance of Gram-negative bacterium cTPY13 was significantly lower (p < 0.05). In addition, Fast Expected Maximum Microbial Source Tracing (FEAST) analysis showed that the contribution of female rabbits' gut microbiota to the colonization of offspring rabbits' gut microbiota was significantly higher than that of unrelated rabbits' gut microbiota (p < 0.05). The contribution of female rabbits' gut microbiota to the colonization of gut microbiota in 0-day-old rabbits was significantly higher than that to the colonization of gut microbiota in the 10- and 60-day-old rabbits (p < 0.05). Finally, the correlation between gut microbiota composition and body weight of 60-day-old rabbits was analyzed based on a mixed linear model, and six ASVs significantly affecting body weight were screened. The above results provide important theoretical and practical guidance for maintaining gut health, improving growth and development performance, and feeding formulation in rabbits. [ABSTRACT FROM AUTHOR]

Featured Application: Postbiotic, Mundticin-like substance EM 41/3 looks as a promising additive for health status sustainability in animal husbandry. Nowadays, the use of probiotics, postbiotics, parabiotics and other beneficial natural substances dominate health status sustainability in animals. Mundticin-like substance EM 41/3 (MLS) is a thermo-stable substance of proteinaceous character—a bacteriocin (postbiotic) with a broad antimicrobial (inhibitory) spectrum produced by the horse-derived strain Enterococcus mundtii EM 41/3. The aim of this study was to find if this new one bacteriocin can have an effect and/or a similar effect in rabbits as was achieved previously by using enterocins detected and characterized in our laboratory. In addition, these tests have been conducted to secure the health of rabbits as food-derived animals. Moreover, the aim of this study was also to test the effect of MLS EM 41/3 in broiler rabbits as an animal model before its use in horses. The parameters tested were analyzed by validated methods. Administration of MLS EM 41/3 lead to a significant increase in the non-specific immunity parameter (phagocytic activity, p < 0.05), while microbiota in feces, caecum and appendix were not negatively influenced. However, the total enterococcal count was significantly increased in the EG compared to the CG (p < 0.001). Application of MLS EM 41/3 did not have a negative impact on blood biochemistry. No oxidative stress was noted. However, higher growth parameters were noted and also hydrolytic activity was increased after MLS EM 41/3 application. Finally, to check physico-chemical parameters, no negative influence on rabbit meat quality was noted. Postbiotic MLS EM 41/3 administration seems to be a contributing factor in the feeding strategy for animal health status sustainability. [ABSTRACT FROM AUTHOR]

The article discusses a study conducted by Shah and colleagues on the genetic characteristics and phylogeographic dynamics of lagoviruses. The study aimed to provide an update on the evolutionary history of lagoviruses and their emergence and re-emergence. However, the authors of this response argue that the methodologies used in the analysis were inappropriate and led to incorrect and misleading inferences. They highlight the importance of considering recombination in phylogenetic analysis and criticize the conclusions drawn by Shah et al. They also point out errors in the classification system proposed by Shah et al. and suggest that their analysis will only generate confusion in the already complex classification of lagoviruses. [Extracted from the article]

The article is a response to a comment made by Abrantes et al. on a previously published article about the genetic characteristics and phylogeographic dynamics of lagoviruses. The authors of the response address the questions raised by Abrantes et al. and defend their classification system and methodology. They argue that their classification system is not complicated and that they have used the latest tools and techniques to make it more reliable. They also explain the importance of considering recombination in virus evolution and the need to include full-length genome sequences in their analysis. The authors clarify some misunderstandings and provide additional evidence to support their findings. [Extracted from the article]

Simple Summary: The aim of this study is to provide information about the Pasteurella multocida strains that are responsible for pasteurellosis in rabbits. A total of 114 strains responsible for different lesions in rabbits from industrial farms were investigated. In detail, the serogroup belonging to and the presence of 15 virulence-associated genes were investigated using PCR (polymerase chain reaction) protocols. The tested strains belonged to serogroups A, D, and F. Type A was the most frequently identified and it was prevalently found in strains responsible for rhinitis and pneumonia. Type D was the prevalent capsular type of strains responsible for metritis, while F was identified in strains detected from otitis, mastitis, subcutaneous abscesses, and septicaemia. Among the virulence-associated genes, sodC was found in all tested strains. Gene pfhA was more frequently found in strains belonging to type A than in type D and it was prevalently found in strains responsible for respiratory lesions. In addition, it was found in all F strains, suggesting a strong relationship between pfhA and this capsular type. Conversely, gene tadD was rarely found in strains belonging to capsular type F. Gene fur occurred more frequently in strains belonging to type D and involved with rhinitis. Gene hgbB was detected prevalently in strains responsible for metritis. Additionally, based on the results, the presence of more than 8 virulence-associated genes in the strains seems to increase the risk of the occurrence of severe lesions in the affected rabbits. Nevertheless, the detection of some strains responsible only for rhinitis which were equipped with 10 and 11 virulence-associated genes, seems to emphasise the importance of a gene's expression rather than its presence in strains. Pasteurellosis, a disease caused by Pasteurella multocida, is responsible for economic losses in rabbit industrial farms due to rhinitis, conjunctivitis, pneumonia, metritis, mastitis, orchitis, subcutaneous abscesses, otitis, encephalitis, and septicaemic forms. Although the occurrence of the disease is conditioned by predisposing factors that affect the rabbit immune response, the strains of P. multocida involved in the infection may have a different pathogenic ability. Therefore, typing of strains spread among the rabbits is important to assess their pathogenic potential. The aim of this study is to investigate the P. multocida strains responsible for disease in rabbit industrial farms. A total of 114 strains identified from different lesions were serotyped. Additionally, the presence of virulence-associated genes was investigated using three PCR (polymerase chain reaction) protocols. Capsular type A was prevalently found in strains from respiratory lesions while types D and F in those from metritis, mastitis, and other lesions. Different associations between some virulence-associated genes and both capsular type and lesions found in rabbits were detected. The presence of 8 virulence-associated genes seems to increase the occurrence of metritis. In addition, strains belonging to capsular type A and responsible for respiratory disorders especially, were found equipped with 10 and 11 virulence-associated genes. Nevertheless, the presence of strains responsible only for rhinitis was also detected among the latter, suggesting that the pathogenic ability of the bacteria depends on the expression rather than the presence of a gene. [ABSTRACT FROM AUTHOR]

Rabbit haemorrhagic disease viruses (RHDV) belong to the family Caliciviridae, genus Lagovirus europaeus, genogroup GI, comprising four genotypes GI.1–GI.4, of which the genotypes GI.1 and GI.2 are pathogenic RHD viruses, while the genotypes GI.3 and GI.4 are non-pathogenic RCV (Rabbit calicivirus) viruses. Among the pathogenic genotypes GI.1 and GI.2 of RHD viruses, an antigenic variant of RHDV, named RHDVa—now GI.1a–RHDVa, was distinguished in 1996; and in 2010, a variant of RHDV—named RHDVb, later RHDV2 and now GI.2–RHDV2/b—was described; and recombinants of these viruses were registered. Pathogenic viruses of the genotype GI.1 were the cause of a disease described in 1984 in China in domestic (Oryctolagus (O.) cuniculus domesticus) and wild (O. cuniculus) rabbits, characterised by a very rapid course and a mortality rate of 90–100%, which spread in countries all over the world and which has been defined since 1989 as rabbit haemorrhagic disease. It is now accepted that GI.1–RHDV, including GI.1a–RHDVa, cause the predetermined primary haemorrhagic disease in domestic and wild rabbits, while GI.2–RHDV2/b cause it not only in rabbits, including domestic rabbits' young up to 4 weeks and rabbits immunised with rabbit haemorrhagic disease vaccine, but also in five various species of wild rabbits and seven different species of hares, as well as wild ruminants: mountain muskoxen and European badger. Among these viruses, haemagglutination-positive, doubtful and harmful viruses have been recorded and described and have been shown to form phylogenogroups, immunotypes, haematotypes and pathotypes, which, together with traits that alter and expand their infectious spectrum (rabbit, hare, wild ruminant, badger and various rabbit and hare species), are the determinants of their pathogenicity (infectivity) and immunogenicity and thus shape their virulence. These relationships are the aim of our consideration in this article. [ABSTRACT FROM AUTHOR]

Cysticercus pisiformis is a kind of tapeworm larvae of Taenia pisiformis, which parasitizes the liver envelope, omentum, mesentery, and rectum of rodents such as rabbits. Cysteine protease inhibitors derived from helminth were immunoregulatory molecules of intermediate hosts and had an immunomodulatory function that regulates the production of inflammatory factors. Thus, in the present research, the recombinant Stefin of C. pisiformis was confirmed to have the potential to fight inflammation in LPS-Mediated RAW264.7 murine macrophages. CCK8 test showed that rCpStefin below 50 μg/mL concentration did not affect cellular viability. Moreover, the NO production level determined by the Griess test was decreased. In addition, the secretion levels of IL-1β, IL-6, and TNF-α as measured by ELISA were decreased. Furthermore, it exerted anti-inflammatory activity by decreasing the production of proinflammatory cytokines and proinflammatory mediators, including IL-1β, IL-6, TNF-α, iNOS, and COX-2 at the gene transcription level, as measured by qRT-PCR. Therefore, Type I cystatin derived from C. pisiformis suppresses the LPS-Mediated inflammatory response of the intermediate host and is a potential candidate for the treatment of inflammatory diseases. [ABSTRACT FROM AUTHOR]

Simple Summary: Domestic rabbits are one of the most commonly kept pets with a hypselodontic type of dentition, possessing teeth that grow throughout the animal's life. These animals often show signs of disease in the stomatognathic system. Dental disease can also affect the function of other organs and systems. The most common clinical signs of dental disease in domestic rabbits are the following: lack of appetite, weight loss, apathy, and difficulty chewing and swallowing food. These clinical signs are non-specific, and, therefore, a definitive diagnosis usually requires additional methods, such as an X-ray examination, a CT scan, MRI, and endoscopic examination. The most common dental disease found in this study's animals was malocclusion secondary to abnormal clinical crown abrasion and abnormal tooth growth. In domestic rabbits, osteomyelitis is a common complication of dental abscesses. Computed tomography is an invaluable diagnostic method in the diagnosis of dental disease and secondary diseases of the head area, such as inflammation of the nasal cavities or otitis media, in pet rabbits. Domestic rabbits have teeth that grow throughout the animal's life and are prone to disease. Clinical signs of dental disease in domestic rabbits are non-specific, and, therefore, a definitive diagnosis usually requires additional methods. This study was carried out on a group of 105 domestic rabbits aged 3 to 9 years. In total, 90 domestic rabbits with dental disease visible on CT images and other secondary diseases of the head area qualified for this study. Malocclusion was found in 57 (63.3%). Retrograde elongation of the tooth apices in the mandible was present in 39 (43.3%), and it was present in the maxilla in 48 (53%). Clinical tooth crowns were overgrown in 39 (43%). Dental abscesses were present in 54 (63%). Secondary to the presence of a dental abscess, osteomyelitis was found in 43 (79% of the animals with a dental abscess). Dental inflammatory resorption was found in 36 (40%). Secondary to dental disease, nasal cavity inflammation was found in 18 (20%). Otitis media was present in six (6.7%). The most common dental disease found in this study's animals was malocclusion secondary to abnormal clinical crown abrasion and abnormal tooth growth. In domestic rabbits, osteomyelitis is a common complication of dental abscesses. Computed tomography is an invaluable diagnostic method in the diagnosis of dental disease and secondary diseases of the head area, such as inflammation of the nasal cavities or otitis media, in pet rabbits. [ABSTRACT FROM AUTHOR]

Rabbit hemorrhagic disease virus 2 (RHDV2) emerged in the United States in 2018 and has spread in both domestic and wild rabbits nationwide. The virus has a high mortality rate and can spread rapidly once introduced in a rabbit population. Vaccination against RHDV2 provides the best protection against disease and should be considered by all rabbit owners. Here, we investigate the duration of immunity provided by vaccination with the Medgene Platform conditionally licensed commercial vaccine 6 months following the initial series. Rabbits received either the vaccination or a placebo and were challenged with RHDV2 6 months later. All vaccinated rabbits survived challenge whereas 18/19 non-vaccinated controls succumbed to infection within 10 or fewer days post-challenge. These results demonstrate lasting immunity following vaccination with the Medgene RHDV2 vaccine. [ABSTRACT FROM AUTHOR]

Rabbit haemorrhage disease virus 2 (RHDV2) is a highly pathogenic lagovirus that causes lethal disease in rabbits and hares (lagomorphs). Since its first detection in Europe in 2010, RHDV2 has spread worldwide and has been detected in over 35 countries so far. Here, we provide the first detailed report of the detection and subsequent circulation of RHDV2 in New Zealand. RHDV2 was first detected in New Zealand in 2018, with positive samples retrospectively identified in December 2017. Subsequent time-resolved phylogenetic analysis suggested a single introduction into the North Island between March and November 2016. Genetic analysis identified a GI.3P-GI.2 variant supporting a non-Australian origin for the incursion; however, more accurate identification of the source of the incursion remains challenging due to the wide global distribution of the GI.3P-GI.2 variant. Furthermore, our analysis suggests the spread of the virus between the North and South Islands of New Zealand at least twice, dated to mid-2017 and around 2018. Further phylogenetic analysis also revealed a strong phylogeographic pattern. So far, no recombination events with endemic benign New Zealand rabbit caliciviruses have been identified. This study highlights the need for further research and surveillance to monitor the distribution and diversity of lagoviruses in New Zealand and to detect incursions of novel variants. [ABSTRACT FROM AUTHOR]

Rabbit hemorrhagic disease (RHD) is an acute fatal disease caused by the rabbit hemorrhagic disease virus (RHDV). Since the first outbreaks of type 2 RHDV (RHDV2) in April 2020 in China, the persistence of this virus in the rabbit population has caused substantial economic losses in rabbit husbandry. Previous failures in preventing RHDV2 prompted us to further investigate the immune mechanisms underlying the virus's pathogenicity, particularly concerning the spleen, a vital component of the mononuclear phagocyte system (MPS). For this, a previous RHDV2 isolate, CHN/SC2020, was utilized to challenge naive adult rabbits. Then, the splenic transcriptome was determined by RNA-Seq. This study showed that the infected adult rabbits had 3148 differentially expressed genes (DEGs), which were associated with disease, signal transduction, cellular processes, and cytokine signaling categories. Of these, 100 upregulated DEGs were involved in inflammatory factors such as IL1α, IL-6, and IL-8. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that these DEGs were significantly enriched in the cytokine–cytokine receptor interaction signaling pathway, which may play a vital role in CHN/SC2020 infection. At the same time, proinflammatory cytokines and chemokines were significantly increased in the spleen at the late stages of infection. These findings suggested that RHDV2 (CHN/SC2020) might induce dysregulation of the cytokine network and compromise splenic immunity against viral infection, which expanded our understanding of RHDV2 pathogenicity. [ABSTRACT FROM AUTHOR]

Pathogenic lagoviruses (Rabbit hemorrhagic disease virus, RHDV) are widely spread across the world and are used in Australia and New Zealand to control populations of feral European rabbits. The spread of the non-pathogenic lagoviruses, e.g., rabbit calicivirus (RCV), is less well studied as the infection results in no clinical signs. Nonetheless, RCV has important implications for the spread of RHDV and rabbit biocontrol as it can provide varying levels of cross-protection against fatal infection with pathogenic lagoviruses. In Chile, where European rabbits are also an introduced species, myxoma virus was used for localised biocontrol of rabbits in the 1950s. To date, there have been no studies investigating the presence of lagoviruses in the Chilean feral rabbit population. In this study, liver and duodenum rabbit samples from central Chile were tested for the presence of lagoviruses and positive samples were subject to whole RNA sequencing and subsequent data analysis. Phylogenetic analysis revealed a novel RCV variant in duodenal samples that likely originated from European RCVs. Sequencing analysis also detected the presence of a rabbit astrovirus in one of the lagovirus-positive samples. [ABSTRACT FROM AUTHOR]

Simple Summary: Rabbits' participation in Animal-Assisted Interventions is increasing. Rabbits are loveable animals, their handling is quick to learn, their body language is simple to read and they can be easily transported into several institutions from kindergarten to palliative care homes. While the positive impact of interventions on humans is indisputable, there is little available information about the effects on animals themselves. Regularly transporting the rabbits to the place of the intervention can be a source of stress, but we assume that they can be trained, and animals can get used to it by offering feed (hay, carrot and apple) and providing appropriate circumstances. Cortisol hormone secretion is initiated as the response to stress and metabolites of cortisol can be isolated in feces samples by laboratory analysis. Using this method we can obtain adequate information about the experienced stress caused the transportation, without disturbing the animals. According to our results, based on the laboratory analysis, repeated transportation causes a significant rise in stress hormone metabolite levels in feces samples regardless of the offered treats during the transport. Those owners who use rabbits for Animal-Assisted Intervention purposes need to take into account that transportation itself is a stressful experience for the animals. (1) Background: the popularity of rabbits has increased during the last decade and become the third most common companion animal in the EU. Rabbits' participation in Animal-Assisted Interventions (AAIs) is growing. It is highly important to ensure the well-being of the animals in AAIs. Whereas the needs and the advantages of people involved in AAI are becoming more and more evident, the needs of animals are not clearly defined, therefore, it is a great field of inquiry. Animals who are used for AAI need to be transported regularly, which itself might be a source of stress. (2) Methods: the stress of rabbits—caused by transportation—was measured in a non-invasive way: cortisol levels were determined from feces, based on their breakdown products. Eighteen animals were involved in the study. Rabbits experienced a 30 min transportation every second day for two weeks (altogether six times) while 126 samples were collected. (3) Results: rabbits could handle the transportation procedure the first time but subsequently the stress hormone metabolites in feces samples increased regardless of the offered treatments (hay, carrot and apple) during the carriage. (4) Conclusions: those owners who use rabbits for Animal-Assisted Interventions need to take into account that transportation itself is a stressful experience for the animals. [ABSTRACT FROM AUTHOR]

Hereditary hemorrhagic telangiectasia (HHT), also called Rendu–Osler syndrome, is a group of rare genetic diseases characterized by autosomal dominance, multisystemic vascular dysplasia, and age-related penetrance. This includes arteriovenous malformations (AVMs) in the skin, brain, lung, liver, and mucous membranes. The correlations between the phenotype and genotype for HHT are not clear. An HHT Chinese pedigree was recruited. Whole exome sequencing (WES) analysis, Sanger verification, and co-segregation were conducted. Western blotting was performed for monitoring ENG/VEGFα signaling. As a result, a nonsense, heterozygous variant for ENG/CD105: c.G1169A:p. Trp390Ter of the proband with hereditary hemorrhagic telangiectasia type 1 (HHT1) was identified, which co-segregated with the disease in the M666 pedigree. Western blotting found that, compared with the normal levels associated with non-carrier family members, the ENG protein levels in the proband showed approximately a one-half decrease (47.4% decrease), while levels of the VEGFα protein, in the proband, showed approximately a one-quarter decrease (25.6% decrease), implying that ENG haploinsufficiency, displayed in the carrier of this variant, may affect VEGFα expression downregulation. Pearson and Spearman correlation analyses further supported TGFβ/ENG/VEGFα signaling, implying ENG regulation in the blood vessels. Thus, next-generation sequencing including WES should provide an accurate strategy for gene diagnosis, therapy, genetic counseling, and clinical management for rare genetic diseases including that in HHT1 patients. [ABSTRACT FROM AUTHOR]

Lagovirus europaeus/GI.2 causes severe and highly fatal Rabbit Hemorrhagic Disease (RHD). Because of its characteristics, this infection is used as an animal model for acute liver failure (ALF). Apoptosis is one of the key processes underlying ALF and has been described as one of the mechanisms of RHD pathogenesis. Apoptotic cell death has been quite well characterized in infection with different variants of GI.1 strains, but so far, the GI.2 genotype has not been widely studied. In this study, we performed an evaluation of apoptotic cell death in hepatocytes of rabbits infected with Lagovirus europaeus/GI.2. We analyzed the expression of genes involved in apoptotic cell death by real-time PCR and performed immunohistochemical (IHC) assays. We showed a significant increase in the expression of caspase-3 and the proapoptotic Bax and anti-apoptotic Bcl-2 in infected animals. In addition, we recorded increased Bax/Bcl-2 ratios. IHC analyses showed the presence of morphological signs of apoptosis in the hepatocytes of infected rabbits. Our results indicate that caspase-3 and proteins from the Bcl-2 families play a key role in apoptosis induced by Lagovirus europaeus/GI.2 infection. [ABSTRACT FROM AUTHOR]

Chronic myelomonocytic leukemia (CMML) is a hematological neoplasm characterized by monocytosis, splenomegaly, thrombocytopenia, and anemia. Moreover, it is associated with SRSF2 mutations and, rarely, with CSF3R variants. We present the case of an 84-year-old patient with persistent anemia and monocytosis. Due to the presence of dysmorphic granulocytes, monocyte atypia, and myeloid precursors in the peripheral blood cells, the patient was subjected to a bone marrow examination. The diagnosis was consistent with CMML type 2. The Hemocoagulative test showed an increase in fibrinolysis markers. Next-generation targeted sequencing showed TET2 and SRSF2 mutations, along with an unexpected CSF3R germline missense variant, rarely encountered in CMML. The patient started Azacitidine treatment and achieved normal hemostatic process values. In conclusion, we identified a heterozygous germline mutation that, together with TET2 and SRSF2 variants, was responsible for the hemorrhagic manifestation. [ABSTRACT FROM AUTHOR]

The cottontail rabbit papillomavirus (CRPV)-associated VX2 carcinoma of the New Zealand White rabbit serves as a model system for human papillomavirus (HPV)-associated head and neck squamous cell carcinomas (HNSCCs). The aim of this study was to evaluate the tumor-inhibiting effect of RNAi-mediated knockdown of the CRPV oncogenes, E6 and E7, using siRNA-loaded lipopolyplexes (LPPs). VX2-carcinoma-derived cells were cultured for up to 150 passages. In addition, CRPV E6 and E7 oncogenes were transiently expressed in COS-7 cells. Efficiency and safety of LPPs were evaluated in both VX2 cells and the COS-7 cell line. Both of these in vitro CRPV systems were validated and characterized by fluorescence microscopy, Western blot, and RT-qPCR. Efficient knockdown of CRPV E6 and E7 was achieved in VX2 cells and COS-7 cells pretransfected with CRPV E6 and E7 expression vectors. Knockdown of CRPV oncogenes in VX2 cells resulted in reduced viability, migration, and proliferation and led to a G0/G1 block in the cell cycle. CRPV E6 and E7 siRNA-loaded LPPs could represent promising therapeutic agents serving as a paradigm for the treatment of papillomavirus-positive cancers and could be of value for the treatment of CRPV-associated diseases in the rabbit such as papillomas and cancers of the skin. [ABSTRACT FROM AUTHOR]

Rabbit haemorrhagic disease virus (RHDV) is established as a landscape-scale biocontrol that assists the management of invasive European rabbits and their impacts in both Australia and New Zealand. In addition to this, it is also available to land managers to augment rabbit control efforts at a local scale. However, current methods of deploying RHDV to rabbits that rely on the consumption of virus-treated baits can be problematic as rabbits are reluctant to consume bait when there is abundant, green, protein-rich feed available. We ran a suite of interrupted time-series experiments to compare the duration of infectivity of two conventional (carrot and oat baits) and two novel (meat bait and soil burrow spray) methods of deploying RHDV to rabbits. All methods effectively killed exposed rabbits. Soil burrow spray and carrot baits resulted in infection and mortality out to 5 days post their deployment in the field, and meat baits caused infection out to 10 days post their deployment. In contrast, oat baits continued to infect and kill exposed rabbits out to 20 days post deployment. Molecular assays demonstrated high viral loads in deployed baits beyond the duration for which they were infectious or lethal to rabbits. Based on our results, we suggest that the drying of meat baits may create a barrier to effective transmission of RHDV by adult flies within 10 days. We therefore hypothesise that fly larvae production and development on infected tissues is critical to prolonged viral transmission from meat baits, and similarly from carcasses of RHDV mortalities, via mechanical fly vectors. Our study demonstrates that meat baits and soil spray could provide additional virus deployment options that remove the need for rabbits to consume baits at times when they are reluctant to do so. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs, miRs) are a group of small, 17–25 nucleotide, non-coding RNA sequences that, in their mature form, regulate gene expression at the post-transcriptional level. They participate in many physiological and pathological processes in both humans and animals. One such process is viral infection, in which miR-155 participates in innate and adaptive immune responses to a broad range of inflammatory mediators. Recently, the study of microRNA has become an interesting field of research as a potential candidate for biomarkers for various processes and disease. To use miRNAs as potential biomarkers of inflammation in viral diseases of animals and humans, it is necessary to improve their detection and quantification. In a previous study, using reverse transcription real-time quantitative PCR (RT-qPCR), we showed that the expression of ocu-miR-155-5p in liver tissue was significantly higher in rabbits infected with Lagovirus europaeus/Rabbit Hemorrhagic Disease Virus (RHDV) compared to healthy rabbits. The results indicated a role for ocu-miR-155-5p in Lagovirus europaeus/RHDV infection and reflected hepatitis and the impairment/dysfunction of this organ during RHD. MiR-155-5p was, therefore, hypothesized as a potential candidate for a tissue biomarker of inflammation and examined in tissues in Lagovirus europaeus/RHDV infection by dPCR. The objective of the study is the absolute quantification of ocu-miR-155-5p in four tissues (liver, lung, kidney, and spleen) of rabbits infected with Lagovirus europaeus/RHDV by digital PCR, a robust technique for the precise and direct quantification of small amounts of nucleic acids, including miRNAs, without standard curves and external references. The average copy number/µL (copies/µL) of ocu-miRNA-155-5p in rabbits infected with Lagovirus europaeus GI.1a/Rossi in the liver tissue was 12.26 ± 0.14, that in the lung tissue was 48.90 ± 9.23, that in the kidney tissue was 16.92 ± 2.89, and that in the spleen was 25.10 ± 0.90. In contrast, in the tissues of healthy control rabbits, the average number of copies/µL of ocu-miRNA-155-5p was 5.07 ± 1.10 for the liver, 23.52 ± 2.77 for lungs, 8.10 ± 0.86 for kidneys, and 42.12 ± 3.68 for the spleen. The increased expression of ocu-miRNA-155-5p in infected rabbits was demonstrated in the liver (a fold-change of 2.4, p-value = 0.0003), lung (a fold-change of 2.1, p-value = 0.03), and kidneys (a fold-change of 2.1, p-value = 0.01), with a decrease in the spleen (a fold-change of 0.6, p-value = 0.002). In the study of Lagovirus europaeus/RHDV infection and in the context of viral infections, this is the first report that shows the potential use of dPCR for the sensitive and absolute quantification of microRNA-155-5p in tissues during viral infection. We think miR-155-5p may be a potential candidate for a tissue biomarker of inflammation with Lagovirus europaeus/RHDV infection. Our report presents a new path in discovering potential candidates for the tissue biomarkers of inflammation. [ABSTRACT FROM AUTHOR]

Simple Summary: The aim of this study was to evaluate the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Phyto-L (Pro Tech s.r.l.), a commercial product containing organosulfur compounds (OSCs) such as propyl propane thiosulfonate (PTSO) from Allium spp., on 108 enteropathogenic E. coli (EPEC) strains responsible for colibacillosis in rabbits. Bacterial suspensions with a charge of 108 CFU/mL were tested with different concentrations (20, 10, 5, 2.5, 1.25, 0.6, 0.3, and 0.15 μL/mL) of Phyto-L. To evaluate MBC values, bacterial suspensions corresponding to the MIC and above the MIC were plated on Tryptic Soy agar (TSA) without Phyto-L. The MICs of the tested strains corresponded to 1.25 μL/mL (37/108-34.3%), 2.5 μL/mL (70/108-64.8%), and 5 μL/mL (1/108-0.9%). The MBCs were 1.25 μL/mL (15/108-13.9%), 2.5 μL/mL (46/108-42.6%), 5 μL/mL (9/108-8.3%), 10 μL/mL (20/108-18.5%), 20 μL/mL (8/108-7.4%), and higher than 20 μL/mL (10/108-9.3%). Based on the results obtained, Phyto-L has antibacterial activity on EPEC strains. Therefore, in field applications, Phyto-L should be useful in limiting the E. coli load in the rabbit gut, preventing the occurrence of colibacillosis. Moreover, considering that 104–105 CFU/g of feces is the charge of E. coli normally present in the intestinal contents of rabbits under physiological conditions, it is possible that lower dosages than those found in this study may be effective in preventing the disease in rabbit farms. Colibacillosis, caused by enteropathogenic Escherichia coli (EPEC), is one of the most common diseases in rabbit farms, resulting in economic losses due to mortality and decrease in production. Until recently, antimicrobials were used to both treat and prevent disease on livestock farms, leading to the possible risk of antimicrobial resistance (AMR) and the selection of multidrug-resistant (MDR) bacteria. Therefore, interest in alternative control methods, such as the use of natural substances, has increased in the scientific community. The aim of this study was to evaluate the antimicrobial efficacy of Phyto-L (Pro Tech s.r.l.), a product containing organosulfur compounds (OSCs) such as propyl propane thiosulfonate (PTSO) from Allium spp., against 108 strains of E. coli isolated from rabbits with colibacillosis from 19 farms. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Phyto-L were assessed. Bacterial suspensions with a charge of 108 CFU/mL, corresponding to those found in the rabbit gut under pathologic conditions, were tested with different concentrations from 20 to 0.15 μL/mL of Phyto-L. For each strain, the MIC and concentrations above the MIC were plated on Tryptic Soy agar (TSA) without Phyto-L to assess the MBCs. MIC and MBC values ranged from 1.25 to 5 μL/mL and 1.25 to 20 μL/mL, respectively, depending on the strain tested. The data showed an interesting antibacterial activity of Phyto-L against EPEC strains. Therefore, this product could be effective in preventing colibacillosis in field application, especially considering that 104–105 CFU/g of feces is the amount of E. coli usually found in the gut contents of rabbits under physiological condition. [ABSTRACT FROM AUTHOR]

The idea of a common mucosal immune system (CMS) is 50 years old. Its relevance to immune protection at mucosal sites and its potential to modulate the impact of vaccination-induced protection against infection of the airway has been poorly understood. The consequent failure of the current SARS-CoV-2 vaccination to satisfy expectations with respect to prevention of infection, viral transmission, duration of protection, and pattern of clinical protection, led to public health and medical decisions now under review. This review summarises knowledge of the CMS in man, including the powerful role it plays in immune protection and lessons with respect to what can and cannot be achieved by systemic and mucosal vaccination for the prevention of airway infection. The powerful impact in both health and disease of optimising delivery of immune protection using selected isolates from the respiratory microbiome is demonstrated through a review of randomised controlled trials (RCTs) in subjects with chronic airway disease, and in otherwise healthy individuals with risk factors, in whom the idea of mucosal immune resilience is introduced. This review is dedicated to two giants of mucosal immunology: Professors John Bienenstock and Allan Cripps. Their recent deaths are keenly felt by their colleagues and students. [ABSTRACT FROM AUTHOR]

Simple Summary: In this study, multi-omics techniques were used to reveal the effects of actual temperature and humidity index (THI) on intestinal microbe, function and serum metabolism of Ira rabbits. The increase of THI had a significant effect on the intestinal microbial structure of meat rabbits. In response to environmental THI changes, intestinal microorganisms with similar functions were replaced with each other, harmful bacteria were increased, heat-sensitive factors were upregulated, and disease-related biomarkers were increased in serum. In addition, the potential biomarkers of serum metabolism could be predicted with high predictive accuracy. In future production, the rapid detection of biomarkers, which has guiding significance for scientific meat rabbit production, can be attempted to determine whether the use of THI in the small environments of meat rabbits is suitable. This study investigates the effects of different THI values on growth performance, intestinal microbes, and serum metabolism in meat rabbits. The results showed that there were significant differences in THI in different location regions of the rabbit house. The high-THI group (HG) could significantly reduce average daily gain and average daily feed intake in Ira rabbits (p < 0.05). The low-THI group (LG) significantly increased the relative abundance of Blautia (p < 0.05). The HG significantly increased the relative abundance of Lachnospiraceae NK4A136 group and reduced bacterial community interaction (p < 0.05). The cytokine–cytokine receptor interactions, nuclear factor kappa B signaling pathway, and toll-like receptor signaling pathway in each rabbit's gut were activated when the THI was 26.14 (p < 0.05). Metabolic pathways such as the phenylalanine, tyrosine, and tryptophan biosynthesis and phenylalanine metabolisms were activated when the THI was 27.25 (p < 0.05). Meanwhile, the TRPV3 and NGF genes that were associated with heat sensitivity were significantly upregulated (p < 0.05). In addition, five metabolites were found to be able to predict THI levels in the environment with an accuracy of 91.7%. In summary, a THI of 26.14 is more suitable for the growth of meat rabbits than a THI of 27.25, providing a reference for the efficient feeding of meat rabbits. [ABSTRACT FROM AUTHOR]

A Japanese rabbit hepatitis E virus (HEV) strain, JP-59, has been identified in a feral rabbit. When this virus was transmitted to a Japanese white rabbit, it caused persistent HEV infection. The JP-59 strain shares an <87.5% nucleotide sequence identity with other rabbit HEV strains. Herein, to isolate JP-59 by cell culture, we used a 10% stool suspension recovered from a JP-59-infected Japanese white rabbit and contained 1.1 × 107 copies/mL of the viral RNA and using it to infect a human hepatocarcinoma cell line, PLC/PRF/5. No sign of virus replication was observed. Although long-term virus replication was observed in PLC/PRF/5 cells inoculated with the concentrated and purified JP-59 containing a high titer of viral RNA (5.1 × 108 copies/mL), the viral RNA of JP-59c that was recovered from the cell culture supernatants was <7.1 × 104 copies/mL during the experiment. The JP-59c strain did not infect PLC/PRF/5 cells, but its intravenous inoculation caused persistent infection in rabbits. The nucleotide sequence analyses of the virus genomes demonstrated that a total of 18 nucleotide changes accompanying three amino acid mutations occurred in the strain JP-59c compared to the original strain JP-59. These results indicate that a high viral RNA titer was required for JP-59 to infect PLC/PRF/5 cells, but its replication capability was extremely low. In addition, the ability of rabbit HEVs to multiply in PLC/PRF/5 cells varied depending on the rabbit HEV strains. The investigations of cell lines that are broadly susceptible to rabbit HEV and that allow the efficient propagation of the virus are thus needed. [ABSTRACT FROM AUTHOR]

Vaccines play an important role in maintaining human and animal health worldwide. There is continued demand for effective and safe adjuvants capable of enhancing antigen-specific responses to a target pathogen. Rabbit hemorrhagic disease virus (RHDV) is a highly contagious calicivirus that often induces high mortality rates in rabbits. Herein, we evaluated the activity of an experimental sulfated lactosyl archaeol (SLA) archaeosome adjuvant when incorporated in subunit vaccine formulations targeting RHDV. The subunit antigens consisted of RHDV–CRM197 peptide conjugates or recombinant RHDV2 VP60. SLA was able to enhance antigen-specific antibody titers and cellular responses in mice and rabbits. Three weeks following immunization, antigen-specific antibody levels in rabbits vaccinated with RHDV2 VP60 + SLA were significantly higher than those immunized with antigen alone, with geomean titers of 7393 vs. 117. In addition, the SLA-adjuvanted VP60-based formulations were highly efficacious in a rabbit RHDV2 challenge model with up to 87.5% animals surviving the viral challenge. These findings demonstrate the potential utility of SLA adjuvants in veterinary applications and highlight its activity in different types of mammalian species. [ABSTRACT FROM AUTHOR]

Simple Summary: The emergence of two strains of rabbit haemorrhagic disease (i.e., GI.1 and GI.2) in the1990s and 2010s, respectively, has been the primary determinant for the decline of wild European rabbits within their native range. We compared the impact of both strains on the wild rabbit populations in Spain using national hunting bags. Our findings showed that GI.1 had a greater impact on wild rabbit populations than GI.2. This disparity is likely to be explained by several factors, such as climatic conditions, host resistance improvement, virulence attenuation, and population density, among others. The outbreaks of two strains of rabbit haemorrhagic disease (RHD) (GI.1 and GI.2) in the Iberian Peninsula have caused substantial economic losses in commercial rabbitries and have affected the conservation of rabbit-sensitive predators due to the dramatic decline of their natural populations. However, the assessment of the impact of both RHD strains on wild rabbit populations has been limited to a few small-scale studies. Little is known about the overall impact within its native range. In this study, we described and compared the effects of GI.1 and GI.2 countrywide by using time series of hunting bag data widely available across the country and compared their trend during the first eight years after the first outbreak of GI.1 (i.e., 1998) and GI.2 (i.e., 2011), respectively. We used Gaussian generalised additive models (GAM) with the number of hunted rabbits as the response variable and year as the predictor to evaluate the non-linear temporal dynamics of the population at the national and regional community levels. The first GI.1 caused a population decline of around 53%, affecting most Spanish regional communities where the disease occurred. The positive trend observed after GI.1 in Spain ended with the initial outbreak of GI.2, which did not appear to cause a national population decline. In contrast, we found significant variability in the rabbit population trend among regional communities, where some increased, and others decreased. Such a disparity is unlikely to be explained by a single factor; rather, it appears to result from several factors, such as climatic conditions, host resistance improvement, virulence attenuation, or population density. Our study suggests that a national comprehensive hunting bag series could aid in elucidating the differences in the impact of emerging diseases on a large scale. Future research should focus on national longitudinal serological studies to shed light on the immunological status of rabbit populations in different regions to better understand the evolution of RHD strains and the resistance gained by the wild populations. [ABSTRACT FROM AUTHOR]

Following the arrival of rabbit haemorrhagic disease virus 2 (RHDV2) in Australia, average rabbit population abundances were reduced by 60% between 2014 and 2018 based on monitoring data acquired from 18 sites across Australia. During this period, as the seropositivity to RHDV2 increased, concurrent decreases were observed in the seroprevalence of both the previously circulating RHDV1 and RCVA, a benign endemic rabbit calicivirus. However, the detection of substantial RHDV1 seropositivity in juvenile rabbits suggested that infections were continuing to occur, ruling out the rapid extinction of this variant. Here we investigate whether the co-circulation of two pathogenic RHDV variants was sustained after 2018 and whether the initially observed impact on rabbit abundance was still maintained. We monitored rabbit abundance and seropositivity to RHDV2, RHDV1 and RCVA at six of the initial eighteen sites until the summer of 2022. We observed sustained suppression of rabbit abundance at five of the six sites, with the average population reduction across all six sites being 64%. Across all sites, average RHDV2 seroprevalence remained high, reaching 60–70% in adult rabbits and 30–40% in juvenile rabbits. In contrast, average RHDV1 seroprevalence declined to <3% in adult rabbits and 5–6% in juvenile rabbits. Although seropositivity continued to be detected in a low number of juvenile rabbits, it is unlikely that RHDV1 strains now play a major role in the regulation of rabbit abundance. In contrast, RCVA seropositivity appears to be reaching an equilibrium with that of RHDV2, with RCVA seroprevalence in the preceding quarter having a strong negative effect on RHDV2 seroprevalence and vice versa, suggesting ongoing co-circulation of these variants. These findings highlight the complex interactions between different calicivirus variants in free-living rabbit populations and demonstrate the changes in interactions over the course of the RHDV2 epizootic as it has moved towards endemicity. While it is encouraging from an Australian perspective to see sustained suppression of rabbit populations in the eight years following the arrival of RHDV2, it is likely that rabbit populations will eventually recover, as has been observed with previous rabbit pathogens. [ABSTRACT FROM AUTHOR]

Streptococcus suis (S. suis) is a bacterial pathogen of pigs that has a major animal health and economic impact on the pig industry. Bovine herpesvirus-4 (BoHV-4) is a new virus-based vaccine vector that has been used for the immunogenic delivery of antigens from a variety of pathogens. In the present study, two recombinant BoHV-4-based vectors were evaluated for their ability to induce immunity and protection against S. suis in a rabbit model. The GMD protein is a fusion protein consisting of multiple dominant B-cell epitopes ((B-cell dominant epitopes of GAPDH, MRP, and DLDH antigens) (BoHV-4/GMD)) and the second suilysin (SLY) (BoHV-4/SLY) from S. suis serotype 2 (SS2). Both GMD and SLY delivered by the BoHV-4 vectors were recognized by sera from SS2-infected rabbits. The vaccination of rabbits with the BoHV-4 vectors induced antibodies against SS2, as well as against additional S. suis serotypes, SS7 and SS9. However, sera from BoHV-4/GMD-vaccinated animals promoted a significant level of phagocytic activity by pulmonary alveolar macrophages (PAMs) against SS2, SS7, and SS9. In contrast, sera from rabbits immunized with BoHV-4/SLY induced PAM phagocytic activity against only SS2. In addition, BoHV-4 vaccines differed in the associated level of protection against lethal SS2 challenge, which ranged from high (71.4%) to low (12.5%) for BoHV-4/GMD and BoHV-4/SLY, respectively. These data suggest BoHV-4/GMD as a promising vaccine candidate against S. suis disease. [ABSTRACT FROM AUTHOR]

Epizootic Hemorrhagic Disease (EHD) of ruminants is a viral pathology that has significant welfare, social, and economic implications. The causative agent, epizootic hemorrhagic disease virus (EHDV), belongs to the Orbivirus genus and leads to significant regional disease outbreaks among livestock and wildlife in North America, Asia, Africa, and Oceania, causing significant morbidity and mortality. During the past decade, this viral disease has become a real threat for countries of the Mediterranean basin, with the recent occurrence of several important outbreaks in livestock. Moreover, the European Union registered the first cases of EHDV ever detected within its territory. Competent vectors involved in viral transmission, Culicoides midges, are expanding its distribution, conceivably due to global climate change. Therefore, livestock and wild ruminants around the globe are at risk for this serious disease. This review provides an overview of current knowledge about EHDV, including changes of distribution and virulence, an examination of different animal models of disease, and a discussion about potential treatments to control the disease. [ABSTRACT FROM AUTHOR]

Simple Summary: Vitamin D is an important hormone that can be acquired through diet or exposure to ultraviolet B (UVB) radiation. Few studies have evaluated the effects of UVB radiation on vitamin D concentrations in the domestic rabbit (Oryctolagus cuniculus); however, initial findings have found they can increase their serum 25-hydroxyvitamin D3 (25-OHD3) following 12 h of artificial UVB exposure. Current husbandry recommendations for rabbits do not include specific UVB lighting requirements. Rabbits are a common pet and research model and are frequently housed indoors without access to natural UVB lighting. Rabbits that are chronically vitamin D deficient may develop mineral deficiencies that can lead to poor calcification of the teeth and skull, predisposing these animals to dental abnormalities, bone infections, and other debilitating diseases. While initial results suggest artificial UVB is positive for rabbits, UVB can also be detrimental to the health of vertebrates. The aim of this study was to determine if shorter-duration UVB exposure could also increase 25-OHD3 concentrations. Rabbits were provided 6 h of artificial UVB daily for 14 days, and there was a significant increase in 25-OHD3 concentrations over time. These findings affirm that rabbits can use short-duration artificial UVB to increase 25-OHD3 concentrations. Vitamin D is an important hormone that can be acquired through diet, exposure to ultraviolet B (UVB) radiation, or a combination of these methods. In domestic rabbits (Oryctolagus cuniculus), both methods appear viable, but there is limited research evaluating the effects of UVB on this species. Previous studies found that 12 h of artificial UVB radiation significantly increased 25-hydroxyvitamin D3 (25-OHD3) concentrations over time. While these findings suggest UVB can be beneficial in rabbits, this form of radiation can also be detrimental to vertebrates. The purpose of this study was to determine if shorter-duration UVB could elicit a similar physiological response in rabbits while minimizing potential negative effects. Six rabbits were used for this pilot study. The baseline serum 25-OHD3 was measured for each rabbit and following 14 days of 6 h/day exposure to artificial UVB, a second 25-OHD3 sample was collected. There was a significant increase (p = 0.001) in serum 25-OHD3 over time (Baseline: 27.7 ± 8.1 nmol/L; Day 14: 79.8 ± 9 nmol/L). This study affirmed that 6 h of UVB produced 25-OHD3 concentrations similar to those found in rabbits exposed to 12 h of UVB. Future studies should continue to determine how the duration of UVB exposure affects 25-OHD3 concentrations. [ABSTRACT FROM AUTHOR]

Rabbit haemorrhagic disease virus (RHDV), European brown hare syndrome virus (EBHSV), rabbit calicivirus (RCV), and hare calicivirus (HaCV) belong to the genus Lagovirus of the Caliciviridae family that causes severe diseases in rabbits and several hare (Lepus) species. Previously, Lagoviruses were classified into two genogroups, e.g., GI (RHDVs and RCVs) and GII (EBHSV and HaCV) based on partial genomes, e.g., VP60 coding sequences. Herein, we provide a robust phylogenetic classification of all the Lagovirus strains based on full-length genomes, grouping all the available 240 strains identified between 1988 and 2021 into four distinct clades, e.g., GI.1 (classical RHDV), GI.2 (RHDV2), HaCV/EBHSV, and RCV, where the GI.1 clade is further classified into four (GI.1a–d) and GI.2 into six sub-clades (GI.2a–f). Moreover, the phylogeographic analysis revealed that the EBHSV and HaCV strains share their ancestor with the GI.1, while the RCV shares with the GI.2. In addition, all 2020–2021 RHDV2 outbreak strains in the USA are connected to the strains from Canada and Germany, while RHDV strains isolated in Australia are connected with the USA-Germany haplotype RHDV strain. Furthermore, we identified six recombination events in the VP60, VP10, and RNA-dependent RNA polymerase (RdRp) coding regions using the full-length genomes. The amino acid variability analysis showed that the variability index exceeded the threshold of 1.00 in the ORF1-encoded polyprotein and ORF2-encoded VP10 protein, respectively, indicating significant amino acid drift with the emergence of new strains. The current study is an update of the phylogenetic and phylogeographic information of Lagoviruses that may be used to map the evolutionary history and provide hints for the genetic basis of their emergence and re-emergence. [ABSTRACT FROM AUTHOR]

Crimean–Congo haemorrhagic fever (CCHF) is a potentially lethal tick-borne viral disease with a wide distribution. In Spain, 12 human cases of CCHF have been confirmed, with four deaths. The diagnosis of CCHF is hampered by the nonspecific symptoms, the high genetic diversity of CCHFV, and the biosafety requirements to manage the virus. RT-qPCR and serological tests are used for diagnosis with limitations. Reverse-transcription loop-mediated isothermal amplification (RT-LAMP) could be an effective alternative in the diagnosis of the disease. However, none of the few RT-LAMP assays developed to date has detected different CCHFV genotypes. Here, we designed a RT-LAMP using a degenerate primer set to compensate for the variability of the CCHFV target sequence. RT-LAMP was performed in colorimetric and real-time tests on RT-qPCR-confirmed CCHF patient samples notified in Spain in 2020 and 2021. Urine from an inpatient was analysed by RT-LAMP for the first time and compared with RT-qPCR. The amplicons obtained by RT-qPCR were sequenced and African III and European V genotypes were identified. RT-LAMP amplified both genotypes and was more sensitive than RT-qPCR in urine samples. We have developed a novel, rapid, specific, and sensitive RT-LAMP test that allows the detection of different CCHFV genotypes in clinical samples. This pan-CCHFV RT-LAMP detected viral RNA for the first time in urine samples. It can be easily performed as a single-tube isothermal colorimetric method on a portable platform in real time and without the need for expensive equipment, thus bringing molecular diagnostics closer to rural or resource-poor areas, where CCHF usually occurs. [ABSTRACT FROM AUTHOR]

Simple Summary: Hepatic coccidiosis is a severe rabbit parasitic disease caused by Eimeria stiedae. It may be fatal for rabbits and leads to important economic losses. The disease is well described in domestic rabbits, but little is known about infection with E. stiedae in wild rabbits. In this study, we investigated (a) whether wild rabbits from the island of Lemnos, Greece, where this species is overpopulated, are infected with E. stiedae and (b) what the effects are of this infection on their liver function. Based on our findings, 13.3% of the wild rabbits included in this study were infected with E. stiedae. Moreover, we observed several alterations in the liver variables in infected individuals compared to the non-infected ones, which were indicative of the negative effects of E. stiedae infection on the liver function of wild rabbits. This study adds to the current knowledge on the pathogens affecting wild rabbits and those circulating in this population on the island of Lemnos, Greece, and shows that E. stiedae infection impairs liver function in wild rabbits. (1) Background: Rabbit hepatic coccidiosis, caused by Eimeria stiedae, is a devastating disease with high morbidity and mortality rates. The disease is well described in rabbits, but little is known about E. stiedae infection in wild rabbits. In this study, we investigated the presence of E. stiedae infection in wild rabbits from the island of Lemnos, Greece, where this species is overpopulated, and the effects of infection on common hepatic biomarkers. (2) Methods: We used liver impression smears to detect the coccidian oocysts, and we defined the liver biochemical profile of the infected individuals. (3) Results: Overall, 13.3% of the liver imprints examined were positive for the presence of coccidial oocysts. The activities of liver enzymes, that is, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and glutamyltransferase (GGT), as well as globulins (GLOB), were increased while the concentrations of albumins (ALB), total proteins (TP) and the albumin to globulin (A/G) ratio were decreased in the infected individuals compared to the non-infected ones. (4) Conclusions: This study adds to the current knowledge on the pathogens affecting wild rabbits and those circulating in this population on the island of Lemnos, Greece. Moreover, we showed that E. stiedae infection exerts pathological effects on the hepatocyte integrity and liver function of wild rabbits, as reflected by the abnormal values of liver injury and dysfunction biomarkers. [ABSTRACT FROM AUTHOR]

Simple Summary: There has been an increase in domestic rabbit ownership in the recent years. The type of dentition of this exotic (non-traditional) animal pet, predisposes it to the development of Acquired Dental Disease (ADD), a disease that affects tooth quality, generates malocclusion, among other dental and non-dental consequences. The aim of this study was to assess the factors that modify the risk for ADD in domestic pet rabbits. For this, 1420 owned rabbits' clinical records from a private practice from 2018 to 2021 were used, recording several variables, including the clinical signs at diagnosis. ADD was detected in 25.4% of the individuals, mostly on their cheek teeth. In addition, age and sex (male) were found to be significant risk factors for ADD. In contrast, a free lifestyle and consuming hay in the diet were protective factors. Understanding the factors that modify the risk of ADD increases the chances of prevention and helps to improve the education of owners and/or guardians of domestic rabbits. Rabbits (Oryctolagus cuniculus) have elodont dentition, a characteristic that predisposes them to the development of Acquired Dental Disease (ADD), which is a multifactorial disease. The aim of this study was to assess the risk factors for ADD in domestic pet rabbits. To do this, a retrospective analysis of 1420 rabbits treated at a private practice during 2018–2021 was performed. For this, several variables were retrieved from clinical records, in addition to signology at the time of diagnosis. ADD was found on 25.4% of rabbits, mostly on their cheek teeth. In addition, age (OR = 1.029; 95% CI = 1.023–1.035; p < 0.001) and sex (male) (OR = 1.591; 95% CI = 1.226–2.064; p < 0.001) were found to be significant risk factors for ADD. In contrast, a free lifestyle (OR = 0.565; 95% CI = 0.362–0.882; p = 0.012) and consuming hay in the diet (OR = 0.323; 95% CI = 0.220–0.473; p < 0.001) were protective factors. In conclusion, ADD has a high prevalence and is usually underdiagnosed, highlighting the need for an exhaustive evaluation of patients during the clinical examination. This study improves our knowledge of ADD and its prevention. [ABSTRACT FROM AUTHOR]

Simple Summary: In rabbits, milk is the primary source of nutrition from early growth to weaning. The ability of the mother rabbit to produce milk, which is also influenced by the maternal genotype, is particularly important in the case of the larger litters. The hormone prolactin is responsible for the initiation and maintenance of lactation and for the synthesis of the major components of milk. Prolactin acts through membrane receptors in target tissues. Point mutations and microsatellites in receptor genes can affect production characteristics. Our aim was to examine the prolactin receptor gene in a wild rabbit (Oryctolagus cuniculus) population with a diverse genetic background. Our hypothesis was that the detected polymorphisms could be associated with milk production. By sequencing the promoter region of the PRLR gene, we detected four point mutations and one microsatellite. Among the genotypes of point mutations in the regulatory region of the PRLR gene, the homozygous genotype and the short repeat of the microsatellite resulted in higher milk production. These could be potential marker candidates for the development of marker-assisted selection. One of the problematic points of rabbit breeding is that the nutritional requirements of the kits are not fully satisfied by the does' milk production from the third week of lactation onwards. The prolactin receptor gene has a significant effect on reproductive processes, and its polymorphisms have been associated with milk production in several species (cattle, goats, sheep, and buffalo). The European wild rabbit (Oryctolagus cuniculus), has a more diverse genetic background compared to domesticated lines. In the course of our study, sequencing of the 1210 bp long segment of the PRLR gene promoter region was accomplished. We detected four point mutations (SNP1-407G > A, SNP2-496G > C, SNP3-926T> and SNP4-973A > C) and one microsatellite at position 574. In our population, the four SNPs were segregated into four genotypes: AACCCCCC, GGGGTTAA, AAGGTTAC, and GGGGTCAC. Our results show that the genotype in the homozygous form is associated with higher milk production (1564.7 ± 444.7 g) compared to the other three genotypes (AACCCCCC 1399.1 ± 326.8 g; GTGACCTT 1403.8 ± 517.1 g; GGGGTCAC 1220.0 ± 666.2 g), and the short microsatellite repeat (167 bp) also coincides with significantly higher milk production (1623.8 ± 525.1 g). These results make the marker-assisted selection possible also for domesticated lines. [ABSTRACT FROM AUTHOR]

As most new medications, Cabotegravir (CAB) was recently approved as an antiretroviral treatment of HIV infection without in-depth safety information on in utero exposure. Although no developmental toxicity in rats and rabbits was reported, recent studies demonstrated that CAB decreases pluripotency of human embryonic stem cells. CAB exposure effects during development were assessed in zebrafish embryos by the Fish Embryo Toxicity test after exposure at subtherapeutic concentrations up to 25× the human Cmax. Larvae behavior was assessed by the light–dark locomotion test. The expression of factors involved in neurogenesis was evaluated by whole-mount in situ hybridization. CAB did not cause gross morphological defects at low doses, although pericardial edema, uninflated swim bladder, decreased heartbeats, growth delay, and decreased hatching rate were observed at the highest concentrations. Decreased locomotion was observed even at the subtherapeutic dose, suggesting alterations of nervous system integrity. This hypothesis was supported by the observation of decreased expression of crucial factors involved in early neuronal differentiation in diencephalic and telencephalic dopaminergic areas, midbrain/hindbrain boundary, and craniofacial ganglia. These findings support CAB effects on neurogenesis in zebrafish embryos and suggest long-term follow-up of exposed infants to provide data on drug safety during pregnancy. [ABSTRACT FROM AUTHOR]

In recent years, several studies have been conducted on Muse cells mainly due to their pluripotency, high tolerance to stress, self-renewal capacity, ability to repair DNA damage and not being tumoral. Additionally, since these stem cells can be isolated from different tissues in the adult organism, obtaining them is not considered an ethical problem, providing an advantage over embryonic stem cells. Regarding their therapeutic potential, few studies have reported clinical applications in the treatment of different diseases, such as aortic aneurysm and chondral injuries in the mouse or acute myocardial infarction in the swine, rabbit, sheep and in humans. This review aims to describe the characterization of Muse cells, show their biological characteristics, explain the differences between Muse cells and mesenchymal stem cells, and present their contribution to the treatment of some diseases. [ABSTRACT FROM AUTHOR]

The European brown hare (Lepus europaeus) is a quite adaptable species, but populations have been decreasing for several decades in different countries, including Germany. To investigate infectious diseases as possible influences on observed population decline in the German federal state Schleswig-Holstein, 118 deceased free-ranging European brown hares were collected between 2017 and 2020 and underwent detailed postmortem examination with extended sampling. Infectious diseases were a major cause of death (34.7%). The number of juveniles found exceeded the adult ones. The main pathomorphological findings were hepatitis (32.8%), pneumonia (22.2%), nephritis (19.1%), liver necrosis (12.9%), and enteritis (40.7%). An unusual main finding was steatitis (20.9%) of unknown origin. Animals were mainly emaciated and showed high infection rates with Eimeria spp. (91.3%) and Trichostrongylus spp. (36.2%). European Brown Hare Syndrome Virus reached an epidemic status with few fatal infections (4.2%) and high seroprevalence (64.9%), whereas the prevalence of Rabbit Haemorrhagic Disease Virus 2 was very low (0.8%) in hares in Schleswig-Holstein. Pathogens such as Yersinia pseudotuberculosis (5.9%), Pasteurella multocida (0.8%), and Staphylococcus aureus (3.4%) only caused sporadic deaths. This study illustrates the wide distribution of various infectious pathogens with high mortality and even zoonotic potential. Infectious diseases need to be considered as an important influence on population dynamics in Schleswig-Holstein. [ABSTRACT FROM AUTHOR]

Since their introduction in 1859, European rabbits (Oryctolagus cuniculus) have had a devastating impact on agricultural production and biodiversity in Australia, with competition and land degradation by rabbits being one of the key threats to agricultural and biodiversity values in Australia. Biocontrol agents, with the most important being the rabbit haemorrhagic disease virus 1 (RHDV1), constitute the most important landscape-scale control strategies for rabbits in Australia. Monitoring field strain dynamics is complex and labour-intensive. Here, using phylodynamic models to analyse the available RHDV molecular data, we aimed to: investigate the epidemiology of various strains, use molecular data to date the emergence of new variants and evaluate whether different strains are outcompeting one another. We determined that the two main pathogenic lagoviruses variants in Australia (RHDV1 and RHDV2) have had similar dynamics since their release, although over different timeframes (substantially shorter for RHDV2). We also found a strong geographic difference in their activities and evidence of overall competition between the two viruses. [ABSTRACT FROM AUTHOR]

Though the first discovery of TRPV6 channel expression in various tissues took place in the early 2000s, reliable tools for its protein detection in various cells and tissues are still missing. Here we show the generation and validation of rabbit polyclonal anti-TRPV6 channel antibodies (rb79–82) against four epitopes of 15 amino acids. Among them, only one antibody, rb79, was capable of detecting the full-length glycosylated form of the TRPV6 channel at around 100 kDa. The generated antibody was shown to be suitable for all in vitro applications, such as immunoblotting, immunoprecipitation, immunocytochemistry, immunofluorescence, etc. One of the most important applications is immunohistochemistry using the paraffin-embedded sections from cancer resection specimens. Using prostate cancer resection specimens, we have confirmed the absence of the TRPV6 protein in both healthy and benign hyperplasia, as well as its expression and correlation to the prostate cancer grades. Thus, the generated rabbit polyclonal anti-TRPV6 channel antibody rb79 is suitable for all in vitro diagnostic applications and particularly for the diagnosis in clinics using paraffin-embedded sections from patients suffering from various diseases and disorders involving the TRPV6 channel. [ABSTRACT FROM AUTHOR]

Simple Summary: China is not only a huge meat rabbit consumer but also the largest meat rabbit producer in the world, contributing a large amount of rabbit meat products to the domestic and foreign markets every year. Therefore, it is important for the domestic and international rabbit meat market to improve rabbit breeding production efficiency and rabbit meat quality based on the good use of domestic feed resources in China. It is well known that dietary amino acid nutrition is of great importance to animal growth. Lysine and methionine are limited in the common domestic rabbit feed sources in China, but they play an important role in rabbit growth. Moreover, different lysine and methionine compositions of the diets respond differently to rabbit growth. Consequently, the search for a better composition of dietary lysine and methionine is the main objective of this study. In recent years, ensuring food security has been an important challenge for the world. It is important to make good use of China's domestic local feed resources to provide safe, stable, efficient, and high-quality rabbit meat products for China and the world. Lysine and methionine are the two most limiting essential amino acids in the rabbit diet. However, little is known about the rational composition of lysine and methionine in rabbit diets and the mechanisms that affect growth and development. Accordingly, in this study, we sought to address this knowledge gap by examining the effects of different compositions of lysine and methionine in rabbit diets. Subsequently, the growth status, nitrogen metabolism, blood biochemical indexes, muscle development, muscle quality, and the growth of satellite cells were evaluated in the animals. The results showed that diets containing 0.80% Lys and 0.40% Met improved average daily weight gain, feed conversion, nitrogen use efficiency, and muscle quality in the rabbits (p < 0.05). Additionally, it altered the amino acid transport potential in muscle by upregulating the expression of the SLC7A10 gene (p < 0.05). Meanwhile, the cell viability and the rate of division and migration of SCs in the 0.80% Lys/0.40 % Met composition group were increased (p < 0.05). SLC38A2 and P–mTOR protein expression was upregulated in the 0.80% lysine/0.40% methionine composition group (p < 0.05). In conclusion, 0.80% Lys/0.40% Met was the most suitable lysine and methionine composition in all tested diets. SLC38A2 acted as an amino acid sensor upstream of mTOR and was involved in the 0.80% Lys/0.40% Met regulation of muscle growth and development, thus implicating the mTOR signaling pathway in these processes. [ABSTRACT FROM AUTHOR]

European brown hare syndrome (EBHS) is one of the main causes of mortality in brown hares (Lepus europaeus) and mountain hares (Lepus timidus) in Europe. Since the mid-1990s, this highly lethal and contagious plague has been widespread in many European countries, contributing to a drastic decline in the number of free-living and farmed hares. A second lagovirus, able to infect some species of hares is rabbit haemorrhagic disease virus 2 (RHDV2; GI.2) recognised in 2010, a new viral emergence of RHDV (GI.1) which is known to be responsible for haemorrhagic disease in rabbits—RHD. The aim of this study was to evaluate the current EBHS epidemiological situation on the basis of the presence of antibodies to European brown hare syndrome virus (EBHSV) and anti-RHDV2 antibodies in sera collected from free-ranging hares in Central and Southeastern Poland in 2020–2021. Additionally, studies on the presence of EBHSV and RHDV2 antigens or their genetic material in the blood and internal organs taken from brown hares between 2014–2021 have been carried out. The results of the serological examination showed nearly 88% of tested blood samples were positive for EBHSV antibodies. No EBHSV was identified in the examined hares using virological and molecular tests. The positive results of EBHS serological studies confirmed the circulation and maintenance of EBHSV in free-living brown hares in Poland. However, no serological, virological or molecular evidence was obtained indicating that the brown hares tested had been in contact with RHDV2. [ABSTRACT FROM AUTHOR]

Simple Summary: Astrovirus is an emerging zoonotic pathogen. Rabbit astrovirus can cause diarrhea in rabbits, which brings great economic losses to the rabbit industry. In this study, a rabbit astrovirus strain named Z317 was successfully isolated from the carcasses of rabbits that suffered from diarrhea, and its whole genome sequence was obtained. In this study, we provide valuable data to help understand the variation and recombination of the virus, and lay a foundation for the prevention and control of the disease and molecular epidemiological research. Rabbit astrovirus (RAstV) is a pathogen that causes diarrhea in rabbits, with high infection rate at various stages, which can often cause secondary or mixed infections with other pathogens, bringing great economic losses to the rabbit industry. In this study, 10 samples were collected from cases of rabbits with diarrhea on a rabbit meat farm in the Shandong area of China. The positive sample for astrovirus detected by RT-PCR was inoculated into an RK 13 cell line. A rabbit astrovirus strain named Z317 was successfully isolated, which produced an obvious cytopathic effect 48 h post-inoculation in the RK 13 cell line. The genome structure of this isolate was studied by high-throughput sequencing, showing that the Z317 strain had the highest similarity with the American strain TN/2208/2010, with 92.43% nucleotide homology, belonging to group MRAstV-23. The basic properties of the Z317 capsid (Cap) protein were analyzed, and 10 liner B cell epitopes were screened with the online biosoft Bepipred 2.0 and SVMTriP, including 445–464, 186–205, 655–674, 88–107, 792–811, 45–64, and 257–276 amino acids. This is the first contribution concerning RAstV genomes in China; more studies are needed to understand the diversity and impact of RAstV on rabbit health. [ABSTRACT FROM AUTHOR]

Simple Summary: The high incidence of stress-associated diseases post rabbit weaning results in great losses threatening the rabbit industry. The increasing emergence of multidrug resistant (MDR) methicillin-resistant Staphylococcus aureus (MRSA) causes life threatening infections worldwide. Thus, the necessity to look inward for natural alternative treatments is now compelling. In this perspective, Nigella sativa extract (NSE) could serve as an effective antibiotic alternate source against MRSA. Herein, NSE was found to possess iin vitro antimicrobial activities against MRSA clinical isolates. Moreover, the synergistic activity between NSE and other antimicrobials was employed to overcome the MRSA resistance. Our findings added new insights for application of NSE in diets of growing rabbits as a growth promoting and an immunostimulant agent, which in turn reduced the high risk associated MRSA infections in growing rabbits. Weaning is the most crucial period associated with increased stress and susceptibility to diseases in rabbits. Methicillin-resistant Staphylococcus aureus (MRSA), a historic emergent pathogen related to post weaning stressors, adversely affects rabbit's growth rate and productive cycle. Since MRSA is rapidly evolving antibiotics resistance, natural products are desperately required to tackle the public health threats posed by antimicrobial resistance. Thus, this study aimed to screen the iin vitro antibacterial activity of Nigella sativa extract (NSE) and its interactions with antibiotics against MRSA isolates. Moreover, 200 weaned rabbits were divided into 4 groups to investigate the iin vivo superiority of NSE graded levels towards growth performance, tight junction integrity, immune responsiveness and resistance against MRSA. Herein, NSE showed promising antimicrobial activities against MRSA isolates from animal (77.8%) and human (64.3%) origins. Additionally, MRSA isolates exposed to NSE became sensitive to all antimicrobials to which they were previously resistant. Our results described that the growth-promoting functions of NSE, especially at higher levels, were supported by elevated activities of digestive linked enzymes. Post-NSE feeding, rabbits' sera mediated bactericidal activities against MRSA. Notably, upregulated expression of occludin, CLDN-1, MUC-2 and JAM-2 genes was noted post NSE supplementation with maximum transcriptional levels in 500 mg/kg NSE fed group. Our data described that NSE constitutively motivated rabbits' immune responses and protected them against MRSA-induced experimental infection. Our results suggest the antimicrobial, growth stimulating and immunomodulation activities of NSE to maximize the capability of rabbits for disease response. [ABSTRACT FROM AUTHOR]

Simple Summary: This work aims to address the use of biomarkers that can provide us with information on the previous nutrition levels of wild rabbits, a keystone species that has drastically reduced in the last few years. Non-esterified fatty acids (NEFA), plasmatic urea nitrogen (PUN), albumin, glutamate and total protein metabolites were analysed. Additionally, we examined the potential of these metabolites as biomarkers for the nutritional and conservation status of European rabbits to further the biological knowledge of this species and contribute to its conservation. European wild rabbit (Oryctolagus cuniculus) populations have drastically reduced, and recently, rabbits have been classed as "endangered" by the IUCN. This animal plays an important ecological role in Mediterranean ecosystems and its introduction could significantly contribute to ecological restoration. Rabbits have high nutrient requirements that apparently cannot be covered in all ecosystems, and there are clues that nutrition can limit their abundance and density. On the other hand, some studies reflect the effects of food availability on the metabolomic status of other animal species, but there are no specific studies on this keystone species. The main aim of this work is to find biomarkers to assess the previous levels of ingestion of European rabbits (Oryctolagus cuniculus). To address this gap, gastric content and blood samples were collected from European rabbits (n = 99) in a Mediterranean area for the analysis of glucose, non-esterified fatty acids (NEFA), plasmatic urea nitrogen (PUN), albumin, glutamate and total protein metabolites. Depending on their previous feed intake (gastric content and the ratio between the gastric content and the weight of the animal), the animals were divided into two groups (lower and normal previous feed intake). Our work shows that the metabolomic profiles of the animals were affected. Levels of glucose (+82%; p = 0.0003), NEFA (−61%; p = 0.0040) and PUN (+139%; p < 0.001) were different in the animals with lower previous feed intake than the animals with normal previous feed intake. This work summarises that metabolic phenotype can be interesting when seeking to discover the limiting nutrients and food availability in diets that could affect the ecological fitness and conservation of European wild rabbits. It is important to mention that in this work, only the effects on six different metabolites have been analysed and more studies are necessary to complement the knowledge of possible metabolites that indicate the level of ingestion in this species and others. These (and new) biomarkers could be used as a tool to provide information about individual or population characteristics that other physiological parameters cannot detect, improving the conservation physiology field. [ABSTRACT FROM AUTHOR]

Lassa virus (LASV) is a zoonotic virus endemic to western Africa that can cause a potentially lethal and hemorrhagic disease, Lassa fever (LF). Survivors suffer a myriad of sequelae, most notably sudden onset sensorineural hearing loss (SNHL), the mechanism of which remains unclear. Unfortunately, studies aiming to identify the mechanism of these sequelae are limited due to the biosafety level 4 (BSL4) requirements of LASV itself. ML29, a reassortant virus proposed as an experimental vaccine candidate against LASV, is potentially an ideal surrogate model of LF in STAT1-/- mice due to similar phenotype in these animals. We intended to better characterize ML29 pathogenesis and potential sequelae in this animal model. Our results indicate that while both CD4 and CD8 T cells are responsible for acute disease in ML29 infection, ML29 induces significant hearing loss in a mechanism independent of either CD4 or CD8 T cells. We believe that this model could provide valuable information for viral-associated hearing loss in general. [ABSTRACT FROM AUTHOR]