Pure bacterial cultures were isolated from diseased snakeheads, Channa maculata (Lacepède), suffering high mortality in a farm in Zhongshan, southern China. Three isolates, namely ZS20100725, ZS20100725-1 and ZS20100725-2, were identified as Aeromonas schubertii. All the isolates showed high 16S rRNA sequence similarities with A. schubertii. The isolates exhibited strong virulence to snakeheads in experimental challenges with LD(50) ranging between 1.4 × 10(4) and 6.4 × 10(6) CFU g(-1). Two of the isolates were positive for haemolysin, elastase, lipase and lecithinase by phenotypic determination, which was further confirmed by PCR amplification of the haemolysin and elastase genes. In sterile liquid medium, the best growth conditions of strain ZS20100725 were 30 °C, pH 7 and 0.5% salinity (w/v). Antibiotic susceptibility tests showed that strain ZS20100725 was susceptible to cefoxitin, cefoperazone and chloramphenicol. Furthermore, histopathology of diseased snakeheads infected with A. schubertii showed necrosis and congestion in liver, kidney and spleen and also damage to the cardiac muscle, intestine and gills.
Background
Litopenaeus vannamei is one of the most important aquaculture shrimps in the world and low temperatures present a serious challenge to its survival, growth, and distribution.
Methods
To investigate their physiological responses during acute cold-stress, L. vannamei were treated under acute cooling conditions from 28 to 13 °C with a cooling rate of 2.5 °C/2 h and were maintained at 13 °C for 12 h. Plasma metabolite concentrations, histological changes, and relative gene expression related to the unfolded protein response (UPR) pathway and apoptosis in the hepatopancreas and the hemocytes of L. vannamei were investigated.
Results
The results revealed that the concentrations of triglycerides, total cholesterol, and total protein in plasma reached their peaks at 23 °C, and then decreased to their minimum values at 13 °C for 12 h. The activity of alkaline phosphatase in the plasma decreased to its lowest level while the activity of alanine aminotransferase increased to its highest level at 13 °C for 12 h. The hepatic tubules became necrotic and the basement membranes were ruptured at 13 °C for 12 h. The gene expression related to UPR and apoptosis in the hepatopancreas and hemocytes was significantly altered by the decrease in the temperature.
Discussion
The results revealed that acute cold-stress caused histological damage in the hepatopancreas of L. vannamei, reducing its immunity. The three UPR pathways were involved in the process of acute cold-stress and the response of activating transcription factor 6 to UPR may be faster and more directthan the IRE1 and PERK pathways.
Haemulidae species are morphologically diversified fishes with wondrous and changeable coloration. These species are prevalent in global tropical, subtropical, and temperate inshore reef areas. Previous morphological classification within Haemulidae and some related families was problematic, and no comprehensive molecular evaluation was conducted on these groups. In this study, we revealed the first molecular phylogenetic analysis involving representative species within Haemulidae and the relevant families (Nemipteridae and Teraponidae). The analysis was performed using both mitochondrial DNA (16S rRNA) and nuclear DNA (TMO-4c4) genes to construct maximum parsimony, maximum likelihood, and Bayesian analysis tree topologies. The molecular phylogeny recovered well-resolved relationships within Haemulidae species and the problematic taxa. In the trees, the Haemulidae species (except for genus Hapalogenys) were divided into two distinct sister lineages, including grunts (Haemulinae) and sweetlips (Plectorhynchinae). Hapalogenys was positioned outside the major Haemulidae tribe and formed an independent group, which challenged the traditional taxonomy that Hapalogenys was classified into Haemulidae. The results did agree with most current studies that Hapalogenys was only distantly related to Haemulidae and could potentially be removed from this family. Additionally, the genus Diagramma was observed to be tightly grouped inside the clade Plectorhinchus, indicating its highly close affinity within Plectorhinchus. Regarding interfamily relationships, the phylogenetic constructions suggested distant relationships within two pairs: between Scolopsis and Haemulidae and between Teraponidae and Haemulidae. Scolopsis was revealed to be highly close with Nemipteridae, and Teraponidae was clustered in an independent group, which was in accordance with most current taxonomic studies.
Genetic variation and population structure of northern snakehead (Channa argus) from eight locations in China were investigated using mitochondrial DNA control region and adjacent regions sequences. Sequence analysis showed that there were 105 haplotypes in 260 individuals, 48 unique haplotypes and 57 shared haplotypes, but no common haplotype shared by all populations. As a whole, the haplotype diversity was high (h=0.989), while the nucleotide diversity was low (π=0.00482). AMOVA analysis detected significant genetic differentiation among all eight populations (FST=0.328, p<0.01) and 66.17% of the total variance was resulted from intra-population differentiation. UPGMA analysis indicated that the eight populations could be divided into four major clusters, which was consistent with that the eight sampled locations were belonged to four isolated river systems. The neutrality and mismatch distribution tests suggested that the eight populations of C. argus in the sampling locations underwent recent population expansion. Among the eight populations, the Erhai Lake population may represent a unique genetic resource and therefore needs to be conserved.
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