Detection and characterization of potentially pathogenic Aeromonas sobria isolated from fish Hypophthalmichthys molitrix (Cypriniformes: Cyprinidae).

The current study focuses on the detection and characterization of potentially pathogenic Aeromonas sobria from fish silver carp (Hypophthalmichthys molitrix). Assessment of clinical, microbiological, pathological and biochemical characteristics of A. sobria were taken into account in order to understand the epidemiology, frequency and occurrence of this infection. Clinically the infected fish (H. molitrix) was observed for various types of symptoms. A total of 33 colonies of A. sobria strain were isolated from 20 cultured H. molitrix, collected from controlled fish pond. Microscopic examination revealed that the strains were rod-shaped, Gram negative bacteria. The revealed percent probability identification of A. sobria from the biochemical characterization in VITEK system was 93% with gram negative (GN) card. The histopathology of Gills caused by this bacterium, A. sobria indicate haemorrhagic gill epithelia and epithelial hyperplasia. Lamelar epithelial hypertrophy and hyperplasia with degenerative changes of the epithelium and hypertrophic epitheliocystis infected cells on gills of H. molitrix were observed during the present study.

Enhanced production of α-amylase by Penicillium chrysogenum in liquid culture by modifying the process parameters.

In this paper, we have assessed the role of changing physicochemical parameters and substrate types on the production of α-amylase enzyme from Penicillium chrysogenum, with a view to determining the optimal conditions required for its maximum production. The findings of this research revealed that, at pH 6 using linseed oil cake as substratum, α-amylase enzyme production was maximum (550.0 U/mL), when the fungi was incubated for 6 days at 30 °C in 0.1 M acetate buffer. Further, reasonably good production of the α-amylase enzyme was also observed at pH 9 with all the experimented carbon sources as substrates. Moreover, statistical analysis, using analysis of variance (ANOVA) carried out to study the impact of different studied parameters on the α-amylase enzyme production revealed that incubation period of 6-18 days is highly significant (p = 0.01) factor in amylotic activity of the P. chrysogenum. Under the researched out optimal conditions, P. chrysogenum is an economically viable option for the industrial and biotechnological production of α-amylase enzyme.

Specificity of DNA binding and methylation by the M.FokI DNA methyltransferase.

The M.FokI adenine-N(6) DNA methyltransferase recognizes the asymmetric DNA sequence GGATG/CATCC. It consists of two domains each containing all motifs characteristic for adenine-N(6) DNA methyltransferases. We have studied the specificity of DNA-methylation by both domains using 27 hemimethylated oligonucleotide substrates containing recognition sites which differ in one or two base pairs from GGATG or CATCC. The N-terminal domain of M.FokI interacts very specifically with GGATG-sequences, because only one of the altered sites is modified. In contrast, the C-terminal domain shows lower specificity. It prefers CATCC-sequences but only two of the 12 star sites (i.e. sites that differ in 1 bp from the recognition site) are not accepted and some star sites are modified with rates reduced only 2-3-fold. In addition, GGATGC- and CGATGC-sites are modified which differ at two positions from CATCC. DNA binding experiments show that the N-terminal domain preferentially binds to hemimethylated GGATG/C(m)ATCC sequences whereas the C-terminal domain binds to DNA with higher affinity but without specificity. Protein-protein interaction assays show that both domains of M.FokI are in contact with each other. However, several DNA-binding experiments demonstrate that DNA-binding of both domains is mutually exclusive in full-length M.FokI and both domains do not functionally influence each other. The implications of these results on the molecular evolution of type IIS restriction/modification systems are discussed.