The arrival, establishment and spread of a highly virulent Edwardsiella ictaluri strain in farmed tilapia, Oreochr omis spp.

Edwardsiella ictaluri is an emerging bacterial pathogen that affects farmed tilapia ( Oreochromis spp.). This study reports the arrival, establishment, and widespread findings of E. ictaluri in farmed tilapia in Vietnam. Among 26 disease outbreaks from 9 provinces in Northern Vietnam during 2019–2021, 19 outbreaks originated from imported seeds, while outbreaks in seven farms were from domestic sources. Clinically sick fish showed the appearance of numerous white spots in visceral organs, and accumulative mortality reached 30%–65%. Twenty-six representative bacterial isolates recovered from 26 disease outbreaks were identified as E. ictaluri based on a combination of phenotypic tests, genus- and species-specific polymerase chain reaction assays, 16S rRNA and gyrB sequencing, and phylogenetic analysis. All isolates harbored the same virulence gene profiles esrC, evpC, ureA-C, eseI, escD, and virD4. Antimicrobial susceptibility tests revealed that 80.8%–100% of isolates were multidrug resistant, with resistance to 4–8 antimicrobials in the groups of penicillin, macrolides, sulfonamides, amphenicols, and glycopeptides. The experimental challenge successfully induced disease that mimicked natural infection. The median lethal doses (LD ) of the tested isolates (n = 4) were 42–61 colony forming units/fish, indicating their extremely high virulence. This emerging pathogen is established and has spread to various geographical locations, causing serious impacts on farmed tilapia in northern Vietnam. It is likely that this pathogen will continue to spread through contaminated stocks (both imported and domestic sources) and persist. Thus, increased awareness, combined with biosecurity measures and emergent vaccination programs is essential to mitigate the negative impact of this emerging disease on the tilapia farming industry.

Hoạt tính kháng oxy hóa và kháng khuẩn của cao chiết ethanol từ lá Cóc trắng (Lumnitzera racemosa Willd)

Cóc trắng hay còn gọi Cọc vàng (Lumnitzera racemosa) là một loài thực vật ngập mặn với nhiều dược tính quý. Tuy nhiên, các nghiên cứu về loài cây này không nhiều, đặc biệt ở Việt Nam. Trong nghiên cứu này, thành phần hóa học cũng như hoạt tính kháng oxy hóa in vitro và hoạt tính kháng khuẩn gây bệnh trên thủy sản của cao chiết Cóc trắng đã được khảo sát. Kết quả cho thấy cao ethanol Cóc trắng có hàm lượng polyphenol và flavonid tổng được xác định lần lượt là 138,532 mg GAE/g; 182,014 mg QE/g. Kết quả khảo sát hoạt tính kháng oxy hóa cho thấy cao ethanol Cóc trắng cho hoạt tính mạnh nhất trên phương pháp thử nghiệm là ABTS●+(IC50=20,461 μg/mL), DPPH (IC50=81,734 μg/mL) và TAC (OD0,5= 86,943 μg/mL), sau đó là năng lực khử sắt (OD0,5=113,108 μg/mL). Cao chiết Cóc trắng thể hiện hoạt tính kháng đối với 4 dòng vi khuẩn Aeromonas hydrophila, Aeromonas dhakensis, Edwardsiella ictaluri, Streptococcus agalactiae với đường kính kháng khuẩn tương ứng 3,87 mm, 4,93 mm, 4,93 mm, 5,73 mm.

Accuracies of genomic predictions for disease resistance of striped catfish to Edwardsiella ictaluri using artificial intelligence algorithms

Assessments of genomic prediction accuracies using artificial intelligence (AI) algorithms (i.e.,, machine and deep learning methods) are currently not available or very limited in aquaculture species. The principal aim of this study was to examine the predictive performance of these new methods for disease resistance to Edwardsiella ictaluri in a population of striped catfish Pangasianodon hypophthalmus and to make comparisons with four common methods, i.e.,, pedigree-based best linear unbiased prediction (PBLUP), genomic-based best linear unbiased prediction (GBLUP), single-step GBLUP (ssGBLUP) and a non-linear Bayesian approach (notably BayesR). Our analyses using machine learning (i.e.,, ML-KAML) and deep learning (i.e.,, DL-MLP and DL-CNN) together with the four common methods (PBLUP, GBLUP, ssGBLUP and BayesR) were conducted for two main disease resistance traits (i.e.,, survival status coded as 0 and 1 and survival time, i.e.,, days that the animals were still alive after the challenge test) in a pedigree consisting of 560 individual animals (490 offspring and 70 parents) genotyped for 14,154 Single Nucleotide Polymorphism (SNPs). The results using 6,470 SNPs after quality control showed that machine learning methods outperformed PBLUP, GBLUP and ssGBLUP, with the increases in the prediction accuracies for both traits by 9.1–15.4%. However, the prediction accuracies obtained from machine learning methods were comparable to those estimated using BayesR. Imputation of missing genotypes using AlphaFamImpute increased the prediction accuracies by 5.3–19.2% in all the methods and data used. On the other hand, there were insignificant decreases (0.3–5.6%) in the prediction accuracies for both survival status and survival time when multivariate models were used in comparison to univariate analyses. Interestingly, the genomic prediction accuracies based on only highly significant SNPs (P < 0.00001, 318 - 400 SNPs for survival status and 1,362–1,589 SNPs for survival time) were somewhat lower (0.3 to 15.6%) than those obtained from the whole set of 6,470 SNPs. In most of our analyses, the accuracies of genomic prediction were somewhat higher for survival time than survival status (0/1 data). It is concluded that although there are prospects for the application of genomic selection to increase disease resistance to Edwardsiella ictaluri in striped catfish breeding programs, further evaluation of these methods should be made in independent families/populations when more data are accumulated in future generations to avoid possible biases in the genetic parameters estimates and prediction accuracies for the disease resistant traits studied in this population of striped catfish P. hypophthalmus.

Edwardsiella ictaluri, an unusual cause of bacteraemia in a Nigerian child with acute bloody diarrhoea

Enteric septicaemia of catfish was first detected in 1976 as an economically significant disease associated with commercial catfish production. Initially, Edwardsiella ictaluri was a host specific pathogen of catfish species but has also been reported from other hosts other than the catfish such as the zebrafish. E. ictaluri has not been isolated in humans hence it is not a zoonotic infection. There has been no previous report of isolation of this organism in humans. This was a case report of a 5 year old boy who presented with fever, vomiting, passage of bloody stool of 6 days and abdominal pain of a day duration. In the case of this 5 year old boy who presented with features of dysentery, blood culture using BACTEC™ grew E. ictaluri. E. ictaluri may be a pathogen which can infect humans just like another closely related species, Edwardsiella tarda. Although, E. ictaluri has not been reported in humans, could this be the first case? Non availability of diagnostic technique appropriate for its diagnosis may explain the rare incidence of the organism in humans, hence many cases would have been treated without isolating the organism.

Diverse Bacteriophages Infecting the Bacterial Striped Catfish Pathogen Edwardsiella ictaluri

Bacteriophages infecting Edwardsiella ictaluri have been less investigated, although the host bacterium is one of the most important fish pathogens causing enteric septicemia of catfish (ESC). We present here two distinctly novel bacteriophages vB_EiM_PVN06 and vB_EiA_PVN09 infecting Edwardsiella ictaluri E1, with their geographical origins from the Mekong Delta, Vietnam. Bacteriophage vB_EiM_PVN06 native to a mud sample reveals complete differences of biological properties with the phage vB_EiA_PVN09 originated from a viscus of a healthy catfish (Pangasianodon hypophthalmus) cultured in the same area. Morphological analyses combined with genomic data indicate that phage vB_EiM_PVN06 is classified to Myoviridae family and shares high similarity with E. ictaluri phage PEi21 genome, while vB_EiA_PVN09 is a member of Teseptimavirus genus, Autographiviridae family, and mostly closes to phage vB_EcoP_IME390. The vB_EiA_PVN09 is a T7-like bacteriophage, which has been firstly found infecting to E. ictaluri, and host range analysis also evidences for the cross-infection of this phage to Escherichia coli K12 and Escherichia coli DH5α. Together, our research highlights the diversity of bacteriophages infecting the pathogen E. ictaluri and suggests further explorations of lytic phages in environmental niches, to be exploited in feasible strategies of phage therapy in ESC disease control.

Hemolysin Co-regulated Family Proteins Hcp1 and Hcp2 Contribute to Edwardsiella ictaluri Pathogenesis

Edwardsiella ictaluri is a Gram-negative facultative intracellular pathogen causing enteric septicemia of catfish (ESC), a devastating disease resulting in significant economic losses in the U.S. catfish industry. Bacterial secretion systems are involved in many bacteria's virulence, and Type VI Secretion System (T6SS) is a critical apparatus utilized by several pathogenic Gram-negative bacteria. E. ictaluri strain 93–146 genome has a complete T6SS operon with 16 genes, but the roles of these genes are still not explored. In this research, we aimed to understand the roles of two hemolysin co-regulated family proteins, Hcp1 (EvpC) and Hcp2. To achieve this goal, single and double E. ictaluri mutants (EiΔevpC, EiΔhcp2, and EiΔevpCΔhcp2) were generated and characterized. Catfish peritoneal macrophages were able to kill EiΔhcp2 better than EiΔevpC, EiΔevpCΔhcp2, and E. ictaluri wild-type (EiWT). The attachment of EiΔhcp2 and EiΔevpCΔhcp2 to ovary cells significantly decreased compared to EiWT whereas the cell invasion rates of these mutants were the same as that of EiWT. Mutants exposed to normal catfish serum in vitro showed serum resistance. The fish challenges demonstrated that EiΔevpC and EiΔevpCΔhcp2 were attenuated completely and provided excellent protection against EiWT infection in catfish fingerlings. Interestingly, EiΔhcp2 caused higher mortality than that of EiWT in catfish fingerlings, and severe clinical signs were observed. Although fry were more susceptible to vaccination with EiΔevpC and EiΔevpCΔhcp2, their attenuation and protection were significantly higher compared to EiWT and sham groups, respectively. Taken together, our data indicated that evpC (hcp1) is involved in E. ictaluri virulence in catfish while hcp2 is involved in adhesion to epithelial cells and survival inside catfish macrophages.