An Integrated in silico and in vivo study of nucleic acid vaccine against Nocardia seriolae infection in orange-spotted grouper Epinephelus coioides.

Nocardiosis in aquatic animals caused by Nocardia seriolae is a frequently occurring serious infection that has recently spread to many countries. In this study, DNA vaccines containing potential bacterial antigens predicted using the reverse vaccinology approach were developed and evaluated in orange-spotted groupers. In silico analysis indicated that proteins including cholesterol oxidase, ld-transpeptidase, and glycosyl hydroxylase have high immunogenicity and are potential vaccine candidates. In vitro assays revealed the mature and biological configurations of these proteins. Importantly, when compared to a control PBS injection, N. seriolae DNA-based vaccines showed significantly higher expression of IL1ß, IL17, and IFNγ at 1 or 2 days, in line with higher serum antibody production and expression of other cellular immune-related genes, such as MHCI, CD4, and CD8, at 7 days post-immunization. Remarkably, enhanced immune responses and strong protective efficacy against a highly virulent strain of N. seriolae were recorded in DNA vaccine-cholesterol oxidase (pcD::Cho) injected fish, with a relative survival rate of 73.3%. Our results demonstrate that the reverse vaccinology approach is a valid strategy for screening vaccine candidates and pcD::Cho is a promising candidate that can boost both innate and adaptive immune responses and confer considerable protection against N. seriolae infection.

Post-COVID condition: a survey of patients recovered from COVID-19 in Central Vietnam.

INTRODUCTION: Evidence on post-COVID-19 conditions is emerging. This study aims to assess post-COVID conditions and related factors in COVID-19 patients in Central Vietnam. METHODOLOGY: A descriptive cross-sectional study was performed on people who have recovered from COVID-19 at least 2 weeks prior to the online survey. Participants were interviewed face-to-face after 6 and 9 months from the first survey. RESULTS: 53 patients (21.2%) were confirmed to have persistent symptoms, of which, 100% and 94.3% reported prolonged fatigue and full-body weakness respectively. Loss of appetite was reported by 90.6%, while persistent cough, insomnia, and trouble sleeping were reported by 86.3% of patients. Headaches and dyspnea were reported by 69.5% and 56.8% respectively, while other symptoms had lower rates. The prevalence of post-COVID condition showed a statistically significant relationship with the time of infection, duration of illness, treatment place, use of herbal medicine, adherence to the 5K message from Vietnam's Ministry of Health, and daily saltwater mouthwash (p < 0.05). However, the use of medicine and supplements was not related to the post-COVID condition (p > 0.05). After 6 months, 125 participants were interviewed face-to-face, and only 15 people (12.0%) reported having post-COVID symptoms, mainly prolonged fatigue (33.3%) and persistent cough (26.7%). After 9 months, these 15 patients no longer had symptoms related to the post- COVID-19 condition. CONCLUSIONS: The post-COVID condition can persist for several weeks or months, but will mostly be in remission after 6 months, and completely resolve after 9 months from the onset of the infection.

First report on genetic characterization, cell-surface properties and pathogenicity of Lactococcus garvieae, emerging pathogen isolated from cage-cultured cobia (Rachycentron canadum).

The diseased cage-cultured cobia (Rachycentron canadum) displayed clinical signs, haemorrhagic eyes, dorsal darkness and gross pathological lesions, enlargement of spleen and liver. Haemorrhages were found in brain, heart and liver with cumulative mortality rates ranging from 20% to 50%. Extensive congestion in the heart, liver, spleen, kidney and brain was observed histopathologically. Epicarditis and meningitis were also revealed in diseased cobia. All isolates recovered from the organs (liver, spleen, head kidney, posterior kidney, brain and muscle) of cobia were found to be gram-positive, non-motile, ovoid cocci, short-chain-forming (diplococci) and α-haemolytic. The API 32 strep system together with the polymerase chain reaction assay for species-specific primers (pLG1 and pLG2) and the internal transcribed spacer (ITS) region (G1 and L1 primers) confirmed all four selected isolates as Lactococcus garvieae. Partial 16S rDNA nucleotide sequence (~1,100 bp) of one representative L. garvieae isolate AOD109191 (GenBank accession number, MW328528.1) shared 99.9% identities with the 16S rDNA nucleotide sequence of L. garvieae (GenBank accession numbers: MT604790.1). Transmission electron microscopy (TEM) evaluation of one representative L. garvieae isolate (AOD109191) and the results of multiplex PCR did not reveal the presence of the capsular gene cluster (CGC), thus categorizing the isolate as the KG+ phenotype. Capsule staining and TEM observations confirmed the presence of a hyaluronic acid-like capsule, a possible virulence factor in KG+ phenotype L. garvieae isolates. The pathogenic potential of the representative isolate (AOD109191) was assessed through intraperitoneal injection challenges in cobia. The gross lesions and histopathological changes found in experimentally infected cobia were similar to those seen in naturally infected fish. This is the first report that confirms L. garvieae-induced 'warm water lactococcsis' can cause outbreaks of diseases in cage-cultured cobia.

Novel technique for scapulothoracic arthroscopy: A case report of the application of intraoperative 3D imaging.

INTRODUCTION: Snapping shoulder syndrome could be effectively treated with scapulothoracic arthroscopy. The excision of the scapular superomedial corner is assumed to help lower the recurrence rate. However, the amount of resection is still controversial. Furthermore, we lack a technique to measure if the resected amount was adequate based only on arthroscopy evaluation. CASE STUDY PRESENTATION: We describe a 47-year-old man who suffered from severe snapping shoulder syndrome as a consequence of a deformity of the left superomedial scapular corner. The patient had endoscopic bursectomy and superomedial corner resection. Intraoperative three-dimensional CT scans (3D-CT) were used to evaluate the amount of resection. The patient recovered without incident and resumed his usual activities within 30 days following surgery. At the six-month follow-up, there were no recurrent symptoms. CONCLUSION: Intraoperative 3D imaging significantly enhances the safety and efficacy of scapulothoracic arthroscopy. This is a novel technique that, to our knowledge, has not been reported previously in the literature.

Non-oncologic indication for elbow megaprothesis replacement: 2 cases report.

INTRODUCTION: Treatment of elbow bone defects is still a huge challenge in orthopaedic in order to restore the shape and function of the elbow joint. Bone defect reconstruction is very difficult due to biomechanical complexity of the elbow joint and the poor coverage tissue of this area, so mega-prothesis can be considered the most optimal solution in these cases. CASE REPORT: We present two clinical cases of megaprosthesis elbow replacement for treatment of bone defects caused by sequelae of trauma. There is one case of 3 cm bone defect at proximal ulna and one case of 3 cm bone defect at distal humerus. In the 1st case, the elbow joint is fusioned and the second case, the elbow joint is degenerated totally after 3 previous surgery. We performed total elbow replacement with a customized megaprosthesis for them. The Mayo elbow function assessment scale [1] pre-surgery was poor at 50 points. The average age is 35 years old. The mean post-operative follow-up time was 14 months. Range of elbow flexed motion was 135 degrees, both patients were maximally extension, the forearm pronation and supination were 90 and 75 degrees, respectively. The Mayo score is very good with 97,5 points. Both patients were completely satisfied with the postoperative results. CONCLUSION: Our results show that megaprosthesis elbow replacement is a very effective option for cases large elbow bone defects due to trauma sequelae. However, careful preoperative preparation is required for the best outcome.

The moonlighting protein fructose 1,6-bisphosphate aldolase as a potential vaccine candidate against Photobacterium damselae subsp. piscicida in Asian sea bass (Lates calcarifer).

Vaccination is the most effective, safe, and environmentally friendly method to prevent the outbreak of Photobacterium damselae subsp. piscicida (Phdp), a dangerous pathogen in aquaculture worldwide. Here, recombinant proteins of catalase, superoxide dismutase, isocitrate dehydrogenase, fructose 1,6-bisphosphate aldolase (Fba), and a mixture of all four proteins were investigated for their immunoprotective effects against photobacteriosis in Asian sea bass (Lates calcarifer). After immunization, experimental fish showed an increase in specific antibody levels and lysozyme activities, especially the Fba group. After a lethal challenge with Phdp strain AOD105021, the Fba group achieved the highest relative percentage of survival rate (70.21%) and a significantly lower bacterial load in the spleens than other groups 3 days after infection. The results suggest that Fba is a good candidate for subunit vaccine development against photobacteriosis in fish.

Protective efficacy of four heat-shock proteins as recombinant vaccines against photobacteriosis in Asian seabass (Lates calcarifer).

Photobacterium damselae subsp. piscicida (Phdp) is the causative agent of photobacteriosis in marine fish and is responsible for huge losses to marine aquaculture worldwide. Efforts have been made to develop a vaccine against this disease. Heat-shock proteins (HSPs) are a family of proteins that are ubiquitous in cellular life. Bacteria produce elevated levels of HSPs as a survival strategy when exposed to stressful environments in a host during infection. This group of proteins are also important antigens that can induce both humoral and cellular immune responses. In this study, four HSPs of Phdp, HSP90, HSP33, HSP70, and DnaJ, were selected for cloning and recombinant expression. Western blotting with rabbit anti-Phdp helped identify rHSP70 and rHSP33 as immunogenic proteins. Asian seabass (Lates calcarifer) immunised with rHSP90, rHSP33, rHSP70, and rDnaJ showed 48.28%, 62.07%, 51.72%, and 31.03% relative percent survival, respectively, after being challenged with Phdp strain AOD105021. High expression levels of immune-related genes and high antibody titres were observed in the rHSP33 group, and the sera of this group also exhibited a high level of bactericidal activity against Phdp. Collectively, our results suggest that HSP33 is a potential candidate for vaccine development against Phdp infection.

Genotypic diversity, and molecular and pathogenic characterization of Photobacterium damselae subsp. piscicida isolated from different fish species in Taiwan.

Photobacteriosis, caused by Photobacterium damselae subsp. piscicida (Phdp), is a serious disease in marine fish species worldwide. To date, the epidemiological characterization of this pathogen in Taiwan remains limited. In this study, we collected 39 Phdp isolates obtained from different farmed fish for phenotypic and genotypic analysis. Phenotype bioassays using API-20E and API-20NE systems showed that the Phdp is a homogeneous group. However, genotyping using the pulsed-field gel electrophoresis (PFGE) technique revealed genetic variability among Phdp isolates when 13 and 11 different PFGE band patterns were obtained with SmaI and NotI as restriction enzymes, respectively. Phylogenetic analysis using 16S rDNA and the Fur gene clustered Taiwanese isolates and other species of P. damselae in the same clade. In contrast, the ToxR phylogenetic tree, a powerful discriminatory marker, separated the two subspecies. Furthermore, the virulence-associated genes, AIP56, P55, PDP_0080, Sod and Irp1, were detected from all isolates. Virulence testing with nine representative isolates in cobia (Rachycentron canadum) and Asian sea bass (Lates calcarifer) showed that some were highly pathogenic with 80%-100% mortality rates. This study provides epidemiological data of Phdp infections in farmed fish in Taiwan, which is necessary to develop comprehensive prevention and control strategies for the disease.

Isolation and genetic characterization of Nocardia seriolae from snubnose pompano Trachinotus blochii in Vietnam.

A total of 480 cage-cultured fish were collected from 4 coastal provinces in central Vietnam to investigate the causative agent of nocardiosis. Fish displayed unique characteristics such as paleness and lethargy and exhibited haemorrhages and ulcers on the skin. Prominent white nodules varying in size were observed in the spleen, kidney, and liver. Furthermore, histopathological sections showed typical granulomatous lesions in these organs. Using the Ziehl-Neelsen staining method, isolated bacteria exhibited acid-fast, bead-like filament morphology when cultured in brain-heart infusion medium or Ogawa medium. Phylogenetic analysis of 16S rDNA confirmed that the isolated bacterium was Nocardia seriolae. This study demonstrates for the first time an outbreak of N. seriolae in snubnose pompano in central Vietnam.