Journal of Fish Diseases

Aeromonas veronii (A. veronii) is an opportunistic pathogen of fish–human–livestock, which poses a threat to the development of aquaculture. Based on our previous studies on proteomics and genomics, we found out that the aodp gene may be related to the virulence of A. veronii TH0426. However, aodp gene encodes a hypothetical protein with an unknown function, and its role in A. veronii TH0426 is not clear. Here, we first constructed a mutant strain (△‐aodp) to investigate the functional role of aodp in A. veronii TH0426. Compared with the wild strain A. veronii TH0426, the growth rate of strain △‐aodp was slower and was resistant to neomycin and kanamycin, but sensitive to cephalexin. The swimming and swarming ability of △‐aodp strain decreased, and the pathogenicity to mice decreased by 15.84‐fold. Besides, the activity of caspase‐3 in EPCs infected with △‐aodp strain was 1.49‐fold lower than that of the wild strain. We examined 20 factors closely related to A. veronii virulence, among them 17 genes were down‐regulated as a result of aodp deficiency. This study laid a foundation for further studies on the pathogenesis of A. veronii.

Bacillary necrosis is a problematic disease in farming of Mekong striped catfish (Pangasianodon hypophthalmus). The pathogenic bacterium is Edwardsiella ictaluri, causing numerous white spots in swelled liver, kidney and spleen. An alternative to antibiotic treatment and vaccine is to select for improved genetic resistance to the disease that requires to establish a proper challenge test. Here, four challenge tests of Mekong striped catfish against E. ictaluri are reported proposing 3 days of acclimatization of test fish prior to the challenge, with restricted water level in the test, keeping a temperature of 26°C. In the challenge, cohabitant shedders should be released directly into the test tank and make up around ⅓ of the fish, and bacteria should be added directly to water. The last two experiments, with the highest mortality, suggest that any factor involving the dead cohabitants should be removed and that additional experimentation should focus on bacteria (density) and timing for addition of bacteria to water. Genetic analyses revealed that resistance to bacillary necrosis tested in replicated tanks in the same experiment can be considered the same genetic trait.

The aim was to carry out a joint genetic analysis of survival and harvest body weight, recorded in a growth performance test in Mekong striped catfish (Pangasianodon hypophthalmus), and susceptibility to bacillary necrosis (caused by Edwardsiella ictaluri), recorded in challenge tests. Data were from two challenge tested year‐classes (~6,000 fish in both) that both had growth test data available for survival and body weight (~13,000 fish each year). Data were analysed with a linear tri‐variate sire‐dam model without the common environmental effect because otherwise genetic correlations were estimated with large standard errors. Susceptibility to bacillary necrosis was found weakly genetically correlated to both growth and survival in the growth test, while growth was found with moderate favourable genetic correlation to growth survival. To defend continued challenge testing of striped catfish in Vietnam, a strong genetic relationship needs to be established between bacillary necrosis and survival under a natural disease outbreak. This requires another field test (in addition to the growth test) with siblings, without antibiotic treatment and the cause of death continuously monitored. Meanwhile, the routine challenge testing with the aim to indirectly improve field survival through selection should continue.

The aim of this study was to analyse four cohabitation challenge‐test experiments with Mekong striped catfish (Pangasianodon hypophthalmus) against the bacterium Edwardsiella ictaluri. The data were genetically analysed per experiment by three models: 1) a cross‐sectional linear model; 2) a cross‐sectional threshold model; and 3) a linear survival model, at both 50% mortality (for models 1 and 2) and at the end of the test (for all three models). In two of the experiments (3 and 4) that were carried out in two replicated tanks, the predicted family effects (sum of sire, dam and common environmental effects) in each tank were correlated with the family survival in the other replicated tank (cross‐validation). The heritability estimates of resistance to E. ictaluri infection were ≤ 0.012 with the survival model, and up to 0.135 – 0.220 (50% survival) and 0.085 and 0.174 (endpoint survival) for the cross‐sectional linear and threshold models, respectively. The challenge test should aim for an endpoint survival that ceases naturally at 50%. Then, genetic analysis should be carried out for survival at the endpoint (reflecting susceptibility) with a simple cross‐sectional linear model.

Abstract. The antibacterial effect of two lysozyme variants purified from rainbow trout, Salmo gairdneri Richardson, kidney was investigated as part of a project directed towards increasing the disease resistance of fish by the transgenic technique. Seven bacterial strains from five Gram‐negative species, of which one was considered non‐pathogenic, were examined. One of the rainbow trout lysozymes was surprisingly potent, having substantial antibacterial activity on all strains tested. Hen egg‐white lysozyme was bactericidal only against the one species considered non‐pathogenic. The data suggest that lysozyme does play a role in the disease defence of rainbow trout and that the gene for the most active lysozyme may be suitable for testing the transgenic strategy.

Water in aquaculture systems may contain a high load of microorganisms. Reduction in overall bacterial tank water load improves fish health and growth parameters. In this study, the effect of an increase of overall bacterial load in tank water on carp skin mucus was assessed. Intracellular and released high molecular weight glycoproteins (HMGs) of carp skin mucus were analysed for changes using histological, histochemical and biochemical techniques. Increase of bacterial load did not induce obvious clinical responses in carp, but the skin of exposed carp responded quickly. The amount of skin mucus HMGs isolated increased as well as their total glycosylation. An increased goblet cell number was observed for all carbohydrate stainings, but most clearly for acidic glycoconjugates. A change in the terminal presence of some sugars was also seen. After the initial response of carp, an adaptation to the higher bacterial load in the water appeared to occur as mucins had a higher glycosylation. The changes observed suggest that these skin mucus adaptations are part of a primary defence mechanism of mucosal epithelia, even at a low pathogenic pressure.

An efficient clearance and degradation system may be needed during microbial invasion which otherwise would lead to severe inflammation and eventually death. Non‐specific defence mechanisms in fish play an important role at all stages in infection. The non‐specific humoral defence including proteases, lysins and agglutinins, for example, in mucosal secretion is the first line of defence, whereas mucosal lining cells function as the second barrier against invasion. Blood cells, especially granulocytes and monocytes, may destroy microbes present in the circulation and may function as the third line of defence. Finally, endocytically active cells such as endothelial cells, macrophages and granulocytes in organs and tissues may take up and degrade microbes or microbial products. The endocytic and degradation processes strongly depend on the effectiveness of the reticuloendothelial system which consists of endothelial cells and macrophages that line small blood vessels (e.g. sinusoids and ellipsoids). Potentiation of non‐specific defence mechanisms may occur during microbial invasion, leading to more efficient clearance and destruction of pathogens or other harmful substances. In microbial invasion, an inflammatory response such as elevated production of antimicrobial substances is often encountered. Central cells in the production of antimicrobial substances are macrophages and granulocytes, and microbial products in inflammation may alter the cells function to a more activated state in vivo. Activated cells may enhance their antimicrobial capacity and efficiency by producing higher amounts and more active antimicrobial agents. This review concerns the non‐specific defence system and gives an introduction to some of the known non‐specific humoral substances and their induction/suppression, and provides a more extensive introduction to cytokine research and immunomodulation. Cellular aspects of non‐specific defence, including macrophages and their products, are discussed in the light of their function in the reticuloendothelial system in fish.

Ô nhiễm nước gây ra các thay đổi bệnh lý ở cá. Mô bệnh học, với vai trò là một chỉ báo việc tiếp xúc với các chất gây ô nhiễm, thể hiện một công cụ hữu ích để đánh giá mức độ ô nhiễm, đặc biệt là đối với các ảnh hưởng dưới mức chết và mãn tính. Tuy nhiên, một phương pháp tiêu chuẩn hóa cho việc mô tả và đánh giá các thay đổi mô học, chủ yếu sử dụng trong cá nước ngọt, vẫn còn thiếu. Trong bài báo này, các tác giả hiện tại đề xuất một công cụ tiêu chuẩn để đánh giá các phát hiện mô học có thể áp dụng cho các cơ quan khác nhau. Phương pháp dựa trên hai yếu tố: (1) sự mở rộng của một thay đổi bệnh lý được đánh giá bằng 'giá trị điểm'; và (2) tầm quan trọng bệnh lý của sự thay đổi này được xác định là một 'hệ số quan trọng'. Tổng của các giá trị điểm và các hệ số quan trọng nhân với nhau của tất cả các thay đổi được chẩn đoán dẫn đến các chỉ số khác nhau. Với những chỉ số này, phân tích thống kê có thể được thực hiện. Các phương pháp đánh giá dành cho mang, gan, thận và da được mô tả.