A comparative study on the apparent digestibility of selected feedstuffs in hybrid catfish (Clarias macrocephalus × Clarias gariepinus) and Nile tilapia (Oreochromis niloticus)

The apparent digestibility (AD) of dry matter (DM), organic matter (OM), crude protein (CP) and essential amino acids (EAA) in five selected feedstuffs was evaluated in hybrid catfish and Nile tilapia. The AD of DM, OM and CP in cassava leaf meal (CLM) was lower (P < 0.05) than in groundnut meal (GNM), soybean meal (SBM), sesame husk meal (SSH) and shrimp head meal (SHM). The AD of DM, OM and CP among GNM, SBM, SSH and SHM was not different (P > 0.05). The AD of most EAA was higher in SSH (P < 0.05) than in CLM, GNM, SBM and SHM. The AD of most EAA among CLM, GNM, SBM and SHM was not different (P > 0.05). The AD of DM and OM in CLM was higher (P < 0.05) in hybrid catfish than in Nile tilapia, while there was no difference in the AD of CP in CLM between fish species. The AD of DM, OM, CP and EAA among GNM, SBM, SSH and SHM did not differ between hybrid catfish and Nile tilapia. In conclusion, there were only small differences in the nutritional properties between the selected feedstuffs in both fish species. Most EAA in the selected feedstuffs were equally well utilized by hybrid catfish and Nile tilapia.     

Growth performance and robustness of African Catfish Clarias gariepinus (Burchell) in biofloc‐based nursery production with different stocking densities

The aim of this research is to evaluate the effects of biofloc system application at different fish density on the nursery production performance and the robustness of African catfish Clarias gariepinus (Burchell) against Aeromonas hydrophila and salinity stress. An economic analysis was also performed to determine the most optimum fish density in biofloc‐based African catfish nursery production. African catfish with an average body weight of 0.96 ± 0.04 g and average body length of 4.20 ± 0.46 cm, respectively, were distributed in 12 units of circular plastic‐lined tanks (1,020 L). The experiment consisted of four treatments in triplicates, that is, a control without carbon source addition at a density of 4 fish/L, and three biofloc treatments at different densities, that is, BFT4 (4 fish/L), BFT6 (6 fish/L) and BFT8 (8 fish/L). Tapioca flour was used as the organic carbon source in biofloc systems and was added at an estimated C/N ratio of 10. Housing the fish in biofloc systems resulted in higher fish growth, more efficient feed utilization, higher fish robustness against A. hydrophila infection and salinity stress, as well as higher profitability of nursery production. Increasing the fish density resulted in higher mortality. However, higher number of fish produced and lower feed conversion ratio were observed in the treatments with higher density (6 and 8 fish/L). In conclusion, the application of biofloc technology at a density of 8 fish/L could be recommended to increase the production and profitability of African catfish nursery culture.     

Effect of Indian lotus (Nelumbo nucifera Gaertn.) leaf powder on immune status and disease resistance of Nile tilapia

This study assessed the immune response of Nile tilapia (Oreochromis niloticus L.) after feeding on different levels (0.1%, 0.2% and 0.4%) of dietary Indian lotus (Nelumbo nucifera) leaf powder for 45 days. We evaluated both the nonspecific immune response at the end of the feeding period and the resistance to Aeromonas hydrophila. Exposure to Indian lotus resulted in a significant elevation in serum total globulins, serum lysozyme activity, serum killing percentage and the phagocytic activity (p < 0.05). Total serum protein and albumin showed no remarkable variation between tilapia fed on 0.1% Indian lotus and the control group (p > 0.05). In addition, the relative expressions of immune‐related genes, namely interleukin–1β and tumour necrosis factor–α were significantly up‐regulated in tilapia fed on 0.4% Indian lotus as compared to the control group; their expressions were down‐regulated in the other tested groups (p < 0.05). The survival rate of Nile tilapia postchallenge to A. hydrophila reported a significant and dose‐dependent increase in the Indian lotus‐supplemented groups (p < 0.05). Therefore, dietary incorporation of Indian lotus leaves (0.4%, 0.2% and 0.1%) could strengthen the immunity of Nile tilapia and improve its resistance to A. hydrophila infection. Therefore, Indian lotus leaves could serve as potential feed supplements for Nile tilapia.     

Cage‐pond integration of African catfish (Clarias gariepinus) and Nile tilapia (Oreochromis niloticus) with carps

Cage‐pond integration system is a new model for enhancing productivity of pond aquaculture system. A field trial was conducted using African catfish (Clarias gariepinus) and Nile tilapia (Oreochromis niloticus) in cages and carps in earthen ponds. There were four treatments replicated five times: (1) carps in ponds without cage, (2) tilapia at 30 fish m^?3 in cage and carps in open pond, (3) catfish at 100 fish m^?3 in cage and carps in open pond, (4) tilapia and catfish at 30 and 100 fish m^?3, respectively, in separate cages and carps in open pond. The carps were stocked at 1 fish m^?2. The cage occupied about 3% of the pond area. The caged tilapia and catfish were fed and the control ponds were fertilized. Results showed that the combined extrapolated net yield was significantly higher (P < 0.05) in the catfish, tilapia and carps integration system (9.4 ± 1.6 t ha^?1 year^?1) than in the carp polyculture (3.3 ± 0.7 t ha^?1 year^?1). The net return from the tilapia and carps (6860 US$ ha^?1 year^?1) and catfish, tilapia and carps integration systems (6668 US$ ha^?1 year^?1) was significantly higher than in the carp polyculture (1709 US$ ha^?1 year^?1) (P < 0.05). This experiment demonstrated that the cage‐pond integration of African catfish and Nile tilapia with carps is the best technology to increase production; whereas integration of tilapia and carp for profitability.     

Aeromonas shuberti as a cause of multi‐organ necrosis in internal organs of Nile tilapia,Oreochromis niloticus

A disease with white spots in internal organs of Nile tilapia occurred in Zhanjiang, southern China. Multiple, white nodules, 0.8–2.2 mm in diameter, were scattered throughout the liver, spleen and kidney of diseased fish. Signs of nodules reproduced after artificial infection with the isolated strain. Isolated bacteria were Gram‐negative, facultative anaerobic, motile, short rod‐shaped, with a length of 1.2–2.2 μm. Morphological and biochemical tests, as well as phylogenetic analysis, all strongly indicated that the isolate from tilapia is identical to Aeromonas schubertii (A. schubertii) which temporary named LF1708 strain. Antibiotic sensitivity assays showed the LF1708 is sensitive to 24 of 27 tested antibiotics. Pathogenicity test revealed that the isolate at the dose of 3.75 × 10⁶ CFU/g killed 100% of experimental tilapia within 2 days and the dose of 1 × 10⁷ CFU/g killed 100% of experimental zebrafish within 1 day. Histopathology of diseased tilapia infected with A. schubertii showed numerous necrotic lesions widely distributed in spleen, liver and kidney, and infiltration with a large number of bacteria. To our knowledge, this was the first report that associated A. schubertii with mortality in tilapia.     

Mortality and pathology of hybrid catfish,Clarias macrocephalus (Günther) × Clarias gariepinus (Burchell), associated with Edwardsiella ictaluri infection in southern Thailand

Enteric septicaemia of catfish (ESC) caused by Edwardsiella ictaluri is becoming an increasing problem in aquaculture and has been reported worldwide in a variety of fish species. This study reports ESC in hybrid catfish, Clarias macrocephalus (Günther) × Clarias gariepinus (Burchell), cultured in southern Thailand. The bacteria were identified as E. ictaluri by conventional and rapid identification systems, as well as by genetic and phylogenetic characterization. Analysis of 16S rRNA indicated 100% homology to the 16S rRNA sequence of several E. ictaluri strains in GenBank. Plasmid profiles demonstrated 4.0‐ and 5.6‐kb plasmids, compared with the 4.8‐ and 5.6‐kb plasmids in the US isolates, and representative genes of three of the four known pathogenicity islands of US isolates were present. Serologically, lipopolysaccharide (LPS) purified from the Thai isolates was not recognized by a monoclonal antibody against the LPS of US isolates. Fish experimentally infected with E. ictaluri showed 23–100% mortality within 14 days with a 168‐h LD₅₀ of 6.92 × 10⁷ CFU mL^?1 by immersion and a 96‐h LD₅₀ of 1.58 × 10⁶ CFU fish^?1 by intraperitoneal injection. Examination of tissue sections obtained from both naturally and experimentally infected fish indicated that infection of hybrid catfish with E. ictaluri produced lesions in several organs including liver, kidney, spleen, heart and brain. Histopathology findings included cellular necrosis, focal haemorrhage, infiltration of lymphocytes and multifocal granulomatous inflammation in the infected organs.     

Pond farming of Nile tilapia, Oreochromis niloticus (L.), in northern Cameroon. Adding hand-sexed male tilapia to graze the dense algal blooms in ponds with African catfish, Clarias gariepinus (Burchell)

Four earthen ponds (250 m² each) were stocked each with 250 small catfish (W₀=39g). In treatment A, African catfish. Clarias gariepinus (Burchell), were raised in monoculture, while in treatment B catfish were raised in polyculture with an additional 125 male Nile tilapia, Oreochromis niloticus (L.) (W₀=44g). Feeding of cottonseed cake was at about 4% of catfish body weight day^?1. Daily feed quantities, however, were averaged over all four ponds so that each pond received the same amount of cottonseed cake. Rearing time was 118 days. In treatment A, catfish grew to an average weight of 200g. In treatment B, catfish reached 158g and tilapia 185g, Extrapolated marketable fish production was strikingly similar in all four ponds (around 4.8 t ha^?1 year^?1). No synergistic effect was obtained by stocking microphagous tilapia, although the feeding of cottonseed cake enhanced dense algal blooms in all ponds. Catfish did not appear to exploit the tilapia recruits, as an indirect pathway of algae cropping.     

Natural co‐infection of cultured Nile tilapia Oreochromis niloticus with Aeromonas hydrophila and Gyrodactylus cichlidarum experiencing high mortality during summer

Aeromonas hydrophila and Gyrodactylus cichlidarum are common pathogens that induce significant economic losses in farm‐reared Nile tilapia. Nowadays, the sudden appearance of fish mortalities was exaggerated due to mixed and multiple infections. During summer 2016, mass mortality among earthen pond‐farmed Nile tilapia was reported. Clinico‐pathological, bacteriological and parasitological examinations have been demonstrated. As well, the water quality parameters were assessed. The clinical and histopathological findings of the moribund and recently dead fish were characterized by generalized septicaemic signs. The water quality parameters were significantly elevated over the permissible levels, whereas there was an elevation in nitrite (0.04 mg/L), un‐ionized ammonia (0.8 mg/L), hydrogen sulphide levels (153.1 mg/L) and organic matter content (3.79 mg/L). A. hydrophila was identified based on phenotypic characterization, API 20E features and the homology of 16S rRNA gene sequence analysis. In addition, PCR data confirmed the presence of aerolysin (aerA) and haemolysin (hly) genes in the identified A. hydrophila isolates. Gene sequencing and phylogenetic analysis based on 16S rRNA sequence confirmed that A. hydrophila H/A (accession No. MN726928) of the present study displayed 98%–99% identity with the 16S rRNA gene of A. hydrophila. Furthermore, the monogenetic trematode, G. cichlidarum was identified in the wet mounts from the skin and gills of the examined fish with a high infestation rate. In this context, it was reported that the synergistic co‐infection of A. hydrophila and G. cichlidarum with deteriorated water quality parameters could induce exaggerated fish mortalities during hot weather.     

The effect of nearly closed RAS on the feed intake and growth of Nile tilapia (Oreochromis niloticus), African catfish (Clarias gariepinus) and European eel (Anguilla anguilla)

One of the challenges that Recirculating Aquaculture Systems (RAS) are still facing is the risk that in RAS fish grow less than in flow-through systems due to the accumulation of substances originating from feed, fish or bacteria associated with the water re-use. The present study investigated whether RAS with high and low accumulation levels of these substances affect feed intake and growth of Nile tilapia Oreochromis niloticus, African catfish Clarias gariepinus, and European eel Anguilla Anguilla. One-hundred and twenty individuals of each species were used (start body weights: Nile tilapia 264.8 ± 8.3 g; African catfish 253.2 ± 2.1 g and European eel 66.6 ± 1.3 g). For a period of 39 days, growth and feed intake were compared between high and low accumulation RAS. HIGH accumulation RAS was designed for maximal accumulation of substances in the water by operating the system at nearly-closed conditions (30 L/kg feed/d), using mature biofilters and high feed loads; and (2) LOW accumulation RAS was designed to be a proxy for flow-through systems by operating at high water exchange rates (1500 L/kg feed/d), new biofilters and low feed load. HIGH accumulation RAS induced a reduction in feed intake (42%) and growth (83%) of Nile tilapia, as compared to systems that are a proxy for flow-through conditions. This effect was not observed in European eel and African catfish. The cause of this reduced feed intake and growth rate of Nile tilapia is still unclear and should be addressed in further studies.     

In vivo and in vitro assessment of Lactobacillus acidophilus as probiotic for tilapia (Oreochromis niloticus,Perciformes:Cichlidae) culture improvement

In this study, the use of Lactobacillus acidophilus as a probiotic for Nile tilapia (Oreochromis niloticus) culture was studied. Fish survival and the expression of some genes involved in the immune response were assessed. Diet supplementation with L. acidophilus for 15 days caused a significant increase in fish survival during a challenge with Aeromonas hydrophila and variations in immune response related to IL‐1β and transferrin expression in Nile tilapia spleen and kidney. Moreover, extracellular products (ECPs) of L. acidophilus showed high antibacterial activity against fish pathogens such as A. hydrophila and Streptococcus agalactiae in vitro. It was also observed that viable L. acidophilus was able to disrupt quorum sensing activity in Chromobacterium violaceum.     

Plasmid‐mediated antimicrobial resistance in motile aeromonads from diseased Nile tilapia (Oreochromis niloticus)

For the sustainable farming of tilapia, proper maintenance of their health and adequate treatment for infections at appropriate time are inevitable. The indiscriminate use of antibiotics in aquaculture, as a part of treatment and as growth promoters, accelerates antimicrobial resistance (AMR) among the fish pathogens. In the present study, we have isolated diverse aeromonads from Nile tilapia and studied their antibiogram and plasmid profiling. Aeromonas hydrophila, Aeromonas veronii, Aeromonas sobria, Aeromonas dhakensis, Aeromonas caviae, Aeromonas jandaei and Aeromonas aquatica were isolated from infected tilapia (n = 150), and their Shannon wiener diversity index was calculated as 1.926. A. veronii was found to be the most multiple antibiotic‐resistant pathogen with the MAR index of 0.46, and A. aquatica was noticed as the least resistant isolate. The minimum inhibitory concentration of resistant antibiotics was shown as >256 mcg/ml for most of the isolates. The virulent genes such as aerolysin and hemolysin were identified in all the isolates except A. aquatica. The detection of class 1 integrons, plasmid profiling and plasmid curing studies confirmed that AMR exhibited by most of the Aeromonas species is of plasmid mediated. This challenges the risk of wide spread of AMR among the pathogens and subsequent treatment of the infection.     

Clostridium butyricum as probiotic for promoting growth performance,feed utilization,gut health and microbiota community of tilapia (Oreochromis niloticus × O. aureus)

The feeding trial was conducted to evaluate the potentials of Clostridium butyricum in the diet of tilapia. Fish (~14 g) were fed with basal diet supplemented with 0 (Control), 0.5 (C‐1), 1 (C‐2), 2 (C‐3), 4 (C‐4) and 8 (C‐5) g/kg commercial probiotic‐containing C. butyricum (1.5 × 10⁸ CFU/g) for 8 weeks. The results showed that weight gain significantly increased, and feed conversion ratio decreased in the C‐2, C‐3 and C‐4 groups (p < .05). The protein retention (except C‐1 group), lipid retention and apparent digestibility coefficient (ADC) of dry matter in probiotic supplementation groups were significantly enhanced, and ADC of protein in the C‐4 group was also improved (p < .05). The supplementation of probiotic significantly increased villus height in anterior intestines and reduced the numbers of intestinal Escherichia coli (p < .05). High‐throughput sequencing showed that top three phyla namely Planctomycetes in all probiotic‐containing groups, Proteobacteria in the C‐1 and C‐2 groups and Chloroflexi in the C‐3 group had higher level than the NC group. The cumulative mortality was reduced by dietary probiotic after challenging with Aeromonas hydrophila (p < .05). In conclusion, C. butyricum can be supplemented at 1–2 g/kg feed for promoting the growth, feed utilization, gut health and microbiota of tilapia.     

The impact of elevated water nitrate concentration on physiology,growth and feed intake of African catfish Clarias gariepinus (Burchell 1822)

The nitrate threshold concentration in rearing water of African catfish (Clarias gariepinus) was assessed. Female African catfish with an initial mean (SD) weight of 154.3 (7.5) g were exposed to 0.4 (Control), 1.5, 4.2, 9.7 and 27.0 mM nitrate for 42 days. Mean (SD) plasma concentrations of nitrate increased from 71 (29) to 6623 (921) μM at the highest ambient nitrate level. Mean (SD) plasma nitrite concentration ranged from 1.2 (0.5) to 7.9 (9.0) μM. Haematocrit, plasma concentrations of non‐esterified fatty acids (NEFA), cortisol, glucose, lactate, osmolality, gill morphology and branchial Na⁺/K⁺‐ATPase activity were not affected. Feed intake and specific growth rate were significantly reduced at the highest nitrate concentration. We advise not to exceed a water nitrate concentration of 10 mM (140 mg L^?1 NO₃‐N) to prevent the risk of reduced growth and feed intake in African catfish aquaculture.     

Dietary supplementation with low molecular weight sodium alginate improves growth,haematology, immune reactions and resistance against Aeromonas hydrophila in Clarias gariepinus

A few studies have illustrated the effects of sodium salt derived from alginic acid on different fish species. However, little is known about the effect of sodium alginate on catfish (Clarias gariepinus). Therefore, this study was performed to assess the use of low molecular weight sodium alginate ( LMWSA) in C. gariepinus. A total of 180 apparently healthy C. gariepinus with a mean body weight of 45 g were randomly divided into three equal groups (D1, D2 and D3). D1 the control group received a control diet, while D2 and D3 received 1% and 3% LMWSA, respectively, for 8 weeks. A challenge test against Aeromonas hydrophila was performed on 15 randomly selected catfish for 15 days. At the end of the experiment, catfish in D3 that received a diet of 3% LMWSA showed significant increases in the final body weight, weight gain and thermal‐unit growth coefficient compared with those in D2 and D1. There was a significant decrease in the erythrogram in D1 after the 4‐day pathogen challenge. A leucogram revealed leucocytosis, heterophilia and lymphocytosis in catfish in D2 and D3 compared with those in D1. After the 4‐day challenge, the following changes took place: lysozyme, nitric oxide, phagocytic activity and the respiratory burst were significantly elevated in catfish that received LMWSA and were more pronounced in D3 than in D1. The mortalities of catfish have been stopped after pathogen challenge from 8‐day in D1 and D2 where at 6‐day in D3. Thus, administration of 1% and 3% LMWSA enhances the growth, immune response and resistance of C. gariepinus against A. hydrophila.     

Comparative genomic analysis of five high drug‐resistance Aeromonas hydrophila strains induced by doxycycline in laboratory and nine reference strains in Genbank

Aeromonas hydrophila is an opportunistic pathogen and the leading cause of fatal haemorrhagic septicaemia in fish and shellfish. Doxycycline, one of the second generation tetracyclines, has been used in fish farming to fight against infectious diseases caused by A. hydrophila due to its broad‐spectrum antimicrobial activity and lower cost. However, progressive increase in resistance of Aeromonas strains to doxycycline aroused serious concern. In this report, drug‐resistant A. hydrophilaAH10 strains were induced and selected by using a consecutive batch culture system in Mueller‐Hinton Broth (MHB) supplemented with varying concentrations of doxycycline. Five isolates (AH101‐105) were obtained from the bacterial culture induced by 25 μg/ml doxycycline for drug‐resistance analysis. Minimal inhibitory concentrations (MIC) values of all five isolates were 50 times higher than that of the parental strain AH10. All of them also displayed high‐level resistance to sulphonamides and amides. We sequenced five isolates and performed comparative genomic analysis of these draft genomes with nine A. hydrophila complete genomes from GenBank. Results showed that the pan‐genome of 14 strains contains 4,730 genes, 3,056 genes of which present in all strains. The drug‐resistance genes also showed significant difference in these genomes, which indicated dangers of indiscriminate use of antibiotics in aquaculture and the necessity of understanding the variation of antibiotic resistance of A. hydrophila. Pan‐genome analysis further revealed that no specific SNP (single nucleotide polymorphism) or InDel (insertion and deletion variation) was identified in any functional gene locus among the genomes of AH10 mutated strains, in contrast, significant CNVs (copy number variations) and SV (structure variations) for gene groups were identified in all the mutant genomes.     

Effects of Bacillus subtilis strains and the prebiotic Previda® on growth,immune parameters and susceptibility to Aeromonas hydrophila infection in Nile tilapia,Oreochromis niloticus

This study was conducted to evaluate the individual and combined effects of long‐term feeding of diets containing two probiotic Bacillus subtilis group strains (Aqua NZ and AP193) and the prebiotic Previda^®, a commercial hemicellulose extract, on growth performance, immune parameters and Aeromonas hydrophila susceptibility of juvenile Nile tilapia, Oreochromis niloticus. Nile tilapia of average weight 7.47 ± 0.11 g were fed diets formulated with the probiotics and/or the prebiotic, or a control diet for 8 weeks and, subsequently, challenged with A. hydrophila by intragastric gavage at a dosage of 3.9 × 10⁷ CFU per fish. Fish attained a mean weight of 59.5 ± 0.99 g at the end of the growth period. Under the conditions of the present trial, none of the diets significantly improved mean per cent weight gain (P = 0.70), thermal growth coefficient (P = 0.88) or feed conversion ratio (P = 0.87) of Nile tilapia. Except for the diet containing the prebiotic Previda^® only (P = 0.17), all other diets resulted in significantly higher fish survival compared to the control (P < 0.05). The combined effect of the prebiotic and probiotic strains emerged as the most important diet with respect to mortality reduction. The mean lysozyme and respiratory burst activities did not show any significant differences between treatments and control (P = 0.14 and 0.32, respectively). Thus, these probiotic strains used in this study have the potential to prevent disease due to A. hydrophila in O. niloticus.     

Effects of dietary supplementation of a marine thermotolerant bacterium,Bacillus paralicheniformis SO‐1, on growth performance and immune responses of Nile tilapia,Oreochromis niloticus

This study was carried out to evaluate the effects of dietary supplementation of thermotolerant bacterium on growth and immune responses of Nile tilapia (Oreochromis niloticus). Bacillus paralicheniformis SO‐1 was isolated from marine environments and incorporated into four isonitrogenous (300g/kg crude protein; cp) and isocaloric (18 MJ/kg) diets at four concentrations: 0, 5, 10 and 20 g/kg diet. Each diet was fed to triplicate groups of Nile tilapia (41.5 ± 0.5 g average weight) at a daily rate of 3% of their biomass, three times a day for 50 days. At the end of the feeding trial, the growth rates, feed utilization efficiency (feed conversion ratio, protein efficiency ratio, protein productive value), digestive enzymes (protease, amylase and lipase) activities, immunological response (serum lysozyme activity, phagocytic activity, respiratory burst and superoxide dismutase activity) and the expression of immune‐related genes [interleukin‐1 (IL‐1), interleukin‐4 (IL‐4) and interleukin‐12 (IL‐12)] were determined. Growth rates, digestive enzymes activities and immunological parameters were significantly improved (p < 0.05) with increasing supplemental SO‐1 up to 10 g/kg. However, further increase in bacterial concentration to 20 g/kg lead to significant decline in fish performance and immune response (p < 0.05). The expression of IL‐1, IL‐4 and IL‐12 genes was significantly up‐regulated (p < 0.05) in the liver of Nile tilapia fed SO‐1‐treated diets. This study clearly demonstrated that B. paralicheniformis SO‐1 could be considered as an efficient growth promoter and immune‐stimulating probiotic for farmed Nile tilapia.     

Microsatellite–centromere mapping in walking catfish Clarias macrocephalus (Günther, 1864) using gynogenetic diploids

Thirty new microsatellite loci were isolated from a microsatellite‐enriched genomic library of walking catfish Clarias macrocephalus. The CA motif was the most abundant, although several other motifs were also isolated. Two gynogenetic diploid families, each consisting of a female and 50 offspring, were genotyped at 56 microsatellite loci, including 30 loci developed in the present study and 26 loci reported in the previous study. Overall, 33 anonymous microsatellites derived from genomic library and 11 EST‐linked microsatellites were mapped to their centromeres. High levels of chiasma interference were apparent in the walking catfish genome as indicated by an average frequency of second‐division segregation (y) of 0.643 ± 0.248. Twenty‐six loci (59%) showed a high microsatellite–centromere recombination, with a frequency >0.67, and three loci had recombination frequencies >0.9. This study demonstrated that gene–centromere mapping provided a rapid method for the expansion of the initial linkage map of walking catfish.     

Comparison of Edwardsiella ictaluri isolates from different hosts and geographic origins

The intraspecific variability of E. ictaluri isolates from different origins was investigated. Isolates were recovered from farm‐raised catfish (Ictalurus punctatus) in Mississippi, USA, tilapia (Oreochromis niloticus) cultured in the Western Hemisphere and zebrafish (Danio rerio) propagated in Florida, USA. These isolates were phenotypically homologous and antimicrobial profiles were largely similar. Genetically, isolates possessed differences that could be exploited by repetitive‐sequence‐mediated PCR and gyrB sequence, which identified three distinct E. ictaluri genotypes: one associated with catfish, one from tilapia and a third from zebrafish. Plasmid profiles were also group specific and correlated with rep‐PCR and gyrB sequences. The catfish isolates possessed profiles typical of those described for E. ictaluri isolates; however, plasmids from the zebrafish and tilapia isolates differed in both composition and arrangement. Furthermore, some zebrafish and tilapia isolates were PCR negative for several E. ictaluri virulence factors. Isolates were serologically heterogenous, as serum from a channel catfish exposed to a catfish isolate had reduced antibody activity to tilapia and zebrafish isolates. This work identifies three genetically distinct strains of E. ictaluri from different origins using rep‐PCR, 16S, gyrB and plasmid sequencing, in addition to antimicrobial and serological profiling.