Prevalence and antimicrobial resistance of Salmonella enteritidis and other salmonellas in the eggs and egg-storing trays from retails markets of Coimbatore, South India

A study was conducted to determine the incidence of Salmonella enterica serovar Enteritidis and other Salmonella serovars on eggshell, egg contents and on egg-storing trays. A total of 492 eggs and 82 egg-storing trays were examined over a period of 1 year from different retail outlets of a residential area of Coimbatore city, South India. Salmonella contamination was recorded in 38 of 492 (7.7%) eggs out of which 29 was in eggshell (5.9%) and 9 in egg contents (1.8%). Around 7.5% of the egg-storing trays were also found to be contaminated with Salmonella. Serotyping of the Salmonella strains showed that 89.7% of the strains from eggshell, 100% of the strains from egg contents and 71.4% of the strains from egg-storing trays were Salmonella Enteritidis. Other serovarvars encountered were S. Cerro, S. Molade and S. Mbandaka from eggshell and S. Cerro from egg-storing trays. Seasonal variations in the prevalence pattern were identified with, a higher prevalence during monsoon months followed by post-monsoon and premonsoon. Further examination of the Salmonella strains was carried out by testing their antimicrobial sensitivity against 10 commonly used antimicrobials. Results revealed high prevalence of multiple antimicrobial resistance among these strains suggesting possible prior selection by use of antimicrobials in egg production

Synthesis, characterization and physiochemical information, along with antimicrobial studies of some metal complexes derived from an ON donor semicarbazone ligand

Eight new transition metal complexes of benzaldehyde-N(4)–phenylsemicarbazone have been synthesized and characterized by elemental analyses, molar conductance, electronic and infrared spectral studies. In all the complexes, the semicarbazone is coordinated as neutral bidentate ligand. 1H NMR spectrum of [Zn(HL)2(OAc)2] shows that there is no enolisation of the ligand in the complex. The magnetic susceptibility measurements indicate that Cr(III), Mn(II), Fe(III), Co(II) and Cu(II) complexes are paramagnetic and Ni(II) is diamagnetic. The EPR spectrum of [Mn(HL)2(OAc)2] in DMF solution at 77K shows hyperfine sextet with low intensity forbidden lines lying between each of the two main hyperfine lines. The g values calculated for the [Cu(HL)2SO4] complex in frozen DMF, indicate the presence of unpaired electron in the dx2−y2 orbital. The metal ligand bonding parameters evaluated showed strong in-plane bonding and in-plane bonding. The ligand and complexes were screened for their possible antimicrobial activities.

Molecular characterization of a crustinlike antimicrobial peptide in the giant tiger shrimp, Penaeus monodon, and its expression profile in response to various immunostimulants and challenge with WSSV

A crustinlike antimicrobial peptide from the haemocytes of giant tiger shrimp, Penaeus monodon was partially characterized at the molecular level and phylogenetic analysis was performed. The partial coding sequence of 299 bp and 91 deduced amino acid residues possessed conserved cysteine residues characteristic of the shrimp crustins. Phylogenetic tree and sequence comparison clearly confirmed divergence of this crustinlike AMP from other shrimp crustins. The differential expression of the crustinlike AMP in P. monodon in response to the administration of various immunostimulants viz., two marine yeasts (Candida haemulonii S27 and Candida sake S165) and two bglucan isolates (extracted from C. haemulonii S27 and C. sake S165) were noted during the study. Responses to the application of two grampositive probiotic bacteria (Bacillus MCCB101 and Micrococcus MCCB104) were also observed. The immune profile was recorded preand postchallenge white spot syndrome virus (WSSV) by semiquantitative RTPCR. Expressions of seven WSSV genes were also observed for studying the intensity of viral infection in the experimental animals. The crustinlike AMP was found to be constitutively expressed in the animal and a significant downregulation could be noted postchallenge WSSV. Remarkable downregulation of the gene was observed in the immunostimulant fed animals prechallenge followed by a significant upregulation postchallenge WSSV. Tissuewise expression of crustinlike AMP on administration of C. haemulonii and Bacillus showed maximum transcripts in gill and intestine. The marine yeast, C. haemulonii and the probiotic bacteria, Bacillus were found to enhance the production of crustinlike AMP and confer significant protection to P. monodon against WSSV infection

Molecular characterization of a crustin-like, putative antimicrobial peptide, Fi-crustin, from the Indian white shrimp, Fenneropenaeus indicus

Fish & Shellfish Immunology 28 (2010) 216-220

Antimicrobial activity of chitosan against vibrios from freshwater prawn Macrobrachium rosenbergii larval rearing systems

Chitosan is a biocompatible and biodegradable natural polymer with established antimicrobial properties against specific microorganisms. The present study demonstrates its antibacterial activity against 48 isolates of Vibrio species from prawn larval rearing systems. The antibacterial activity had a positive correlation with the concentration of chitosan. This work opens up avenues for using chitosan as a prophylactic biopolymer for protecting prawn larvae from vibriosis.

Molecular characterization and phylogenetic analysis of a penaeidin-like antimicrobial peptide, Fi-penaeidin from Fenneropenaeus indicus

Antimicrobial peptides (AMPs) play a major role in innate immunity. Penaeidins are a family of AMPs that appear to be expressed in all penaeid shrimps. Penaeidins are composed of an N-terminal proline-rich domain, followed by a C-terminal domain containing six cysteine residues organized in two doublets. This study reports the first penaeidin AMP sequence, Fi-penaeidin (GenBank accession number HM243617) from the Indian white shrimp, Fenneropenaeus indicus. The full length cDNA consists of 186 base pairs encoding 61 amino acidswith an ORF of 42 amino acids and contains a putative signal peptide of 19 amino acids. Comparison of F. indicus penaeidin (Fi-penaeidin) with other known penaeidins showed that it shared maximum similarity with penaeidins of Farfantepenaeus paulensis and Farfantepenaeus subtilis (96% each). Fi-penaeidin has a predicted molecular weight (MW) of 4.478 kDa and theoretical isoelectric point (pI) of 5.3

Characterization of Histone H2A Derived Antimicrobial Peptides, Harriottins, from Sicklefin Chimaera Neoharriotta pinnata (Schnakenbeck, 1931) and Its Evolutionary Divergence with respect to CO1 and Histone H2A

Antimicrobial peptides (AMPs) are humoral innate immune components of fishes that provide protection against pathogenic infections. Histone derived antimicrobial peptides are reported to actively participate in the immune defenses of fishes. Present study deals with identification of putative antimicrobial sequences from the histone H2A of sicklefin chimaera, Neoharriotta pinnata. A 52 amino acid residue termed Harriottin-1, a 40 amino acid Harriottin-2, and a 21 mer Harriottin-3 were identified to possess antimicrobial sequence motif. Physicochemical properties andmolecular structure ofHarriottins are in agreement with the characteristic features of antimicrobial peptides, indicating its potential role in innate immunity of sicklefin chimaera. The histone H2A sequence of sicklefin chimera was found to differ from previously reported histone H2A sequences. Phylogenetic analysis based on histone H2A and cytochrome oxidase subunit-1 (CO1) gene revealed N. pinnata to occupy an intermediate position with respect to invertebrates and vertebrates

A Novel Isoform of the Hepatic Antimicrobial Peptide, Hepcidin (Hepc-CB1), from a Deep-Sea Fish, the Spinyjaw Greeneye Chlorophthalmus bicornis (Norman, 1939): Molecular Characterisation and Phylogeny

Hepcidin is cysteine-rich short peptide of innate immune system of fishes, equipped to perform prevention and proliferation of invading pathogens like bacteria and viruses by limiting iron availability and activating intracellular cascades. Hepcidins are diverse in teleost fishes, due to the varied aquatic environments including exposure to pathogens, oxygenation and iron concentration. In the present study, we report a 87-amino acid (aa) preprohepcidin (Hepc-CB1) with a signal peptide of 24 aa, a prodomain of 39 aa and a bioactive mature peptide of 24 aa from the gill mRNA transcripts of the deep-sea fish spinyjaw greeneye, Chlorophthalmus bicornis. Molecular characterisation and phylogenetic analysis categorised the peptide to HAMP2-like group with a mature peptide of 2.53 kDa; a net positive charge (?3) and capacity to form b-hairpin-like structure configured by 8 conserved cysteines. The present work provides new insight into the mass gene duplication events and adaptive evolution of hepcidin isoforms with respect to environmental influences and positive Darwinian selection. This work reports a novel hepcidin isoform under the group HAMP2 from a nonacanthopterygian deep-sea fish, C. bicornis

Molecular Characterisation and Phylogenetic Analysis of a Novel Isoform of Hepatic Antimicrobial Peptide, Hepcidin (Zc-hepc1), from the Coral Fish Moorish idol, Zanclus cornutus (Linnaeus,1758)

Hepcidin is a family of short cysteine-rich antimicrobial peptides (AMPs) participating in various physiological functions with inevitable role in host immune responses. Present study deals with identification and characterisation of a novel hepcidin isoform from coral fish Zanclus cornutus. The 81 amino acid (aa) preprohepcidin obtained from Z. cornutus consists of a hydrophobic aa rich 22 mer signal peptide, a highly variable proregion of 35 aa and a bioactive mature peptide with 8 conserved cysteine residues which contribute to the disulphide back bone. The mature hepcidin, Zc-hepc1 has a theoretical isoelectric point of 7.46, a predicted molecular weight of 2.43 kDa and a net positive charge of ?1. Phylogenetic analysis grouped Z. cornutus hepcidin with HAMP2 group hepcidins confirming the divergent evolution of hepcidin-like peptide in fishes. Zc-hepc1 can attain a b-hairpin-like structure with two antiparallel b-sheets. This is the first report of an AMP from the coral fish Z. cornutus.

Marine Microbial Enzymes

3.4. Lipase (EC-3.1. 1.3) 3.5. Other Known Enzymes 3.6. Extremozymes (Enzymes from extremophiles) 3.7. Recognition of Valuable Extremozymes 4. Enzymes as Tools in Biotechnology 4.1. Restriction Enzymes from Marine Bacteria 4.2. Other Nucleases from Marine Bacteria 4.3. Bacteriolytic Enzyme by Bacteriophage from Seawater 5. Innovations in Enzyme Technology 5.1. Enzyme Engineering 5.2. Immobilization Technology 5.3. Gene Cloning for Marine Enzymes 6. Future Prospects

Catalysis by Enzymes immobilized on Tuned Mesoporous Silica

Mesoporous silica nanoparticles provide a non-invasive and biocompatible delivery platform for a broad range of applications in therapeutics, pharmaceuticals and diagnosis. Additionally, mesoporous silica materials can be synthesized together with other nanomaterials to create new nanocomposites, opening up a wide variety of potential applications. The ready functionalization of silica materials makes them ideal candidates for bioapplications and catalysis. These properties of mesoporous silica like high surface areas, large pore volumes and ordered pore networks allow them for higher loading of drugs or biomolecules. Comparative studies have been made to evaluate the different procedures; much of the research to date has involved quick exploration of new methods and supports. Requirements for different enzymes may vary, and specific conditions may be needed for a particular application of an immobilized enzyme such as a highly rigid support. In this endeavor, mesoporous silica materials having different pore size were synthesized and easily modified with active functional groups and were evaluated for the immobilization of enzymes. In this work, Aspergillus niger glucoamylase, Bovine liver catalase, Candida rugosa lipase were immobilized onto support by adsorption and covalent binding. The structural properties of pure and immobilized supports are analyzed by various characterization techniques and are used for different reactions of industrial applications.

Actinomycete Isolates from Arabian Sea and Bay of Bengal: Biochemical, Molecular and Functional Characterization

The thesis is comprised of seven chapters. Chapter 1 gives a general introduction to marine actinomycetes; Chapter 2 gives an account on the morphological, biochemical and physiological characterization of marine actinomycetes. Comprehensive description of molecular identification and phylogenetic analysis of actinomycetes is dealt with in Chapter 3. The antimicrobial property with special reference to antivibrio activity is described in Chapter 4. Chapter 5 explores the melanin production ability of marine actinomycetes, characterization of melanin and evaluation of its bioactivity. Chapter 6 illustrates the study on chitinolytic Streptomyces as antifungal and insecticidal agents. Summary and Conclusion of the study is presented in Chapter 7, followed by References and Appendices.The present study provides an insight into the various actinomycetes occurring in the sediments of Arabian Sea and Bay of Bengal. Streptomyces was found to be the dominant group followed by Nocardiopsis. Eventhough generic level identification is possible by traditional phenotypic methods, species level identification necessitate a polyphasic approach including both phenotypic and genotypic characterization. Antibiotic production coupled with biogranulation property helped in the effective utilization of the actinomycetes for the control of vibrios. Melanin from Streptomyces bikiniensis was proved to be a promising antioxidant and photoprotectant. Marine actinomycetes were found to be a good source of hydrolytic enzymes and the chitinolytic isolates could be explored as biocontrol agents in terms of antifungal and insecticidal property. The present study explored the potential of marine actinomycetes especially Streptomycetes as a promising source of bioactive molecules for application in aquaculture and pharmaceutical industry.

The Major Digestive Enzymes in Etroplus suratensis and Oreochromis mossambicus: Distribution and Characteristics

The major digestive enzyme activities and digestive indices were compared between Etroplus suratensis and Oreochromis mossambicus. Pepsin - like acid proteases that acts on low pH has been identified all along the digestive tract of both the fishes. Comparatively low alpha amylase activity is shown by the E. suratensis and the enzyme is distributed almost equally throughout the intestinal segments in both the species. Very low alkaline protease activity is found in the stomach of both the fishes and in O. mossambicus, the enzyme activity diminishes extensively towards the posterior portion of the intestine whereas in E. suratensis the activity increases towards the posterior part. The present study showed that lipase is one of the prominent digestive enzymes in O. mossambicus with a remarkable specific activity throughout the digestive tract than that of E. suratensis .It has been noted that O. mossambicus has a higher values for digestive somatic index, hepato somatic index, intestinal coefficient and gut Vs standard length ratio than that of E. suratensis indicating its higher digestive and metabolic capabilities. The early maturity and fast growth of O. mossambicus can be explained by their enhanced digestive indices. The compa ratively low activities of acid protease, amylase, lipase and total alkaline protease of E. suratensis revealed poor digestive capacity than that of O. mossambicus

Characterization and applications of two protease enzymes obtained by culture dependent and independent approaches from mangrove sediments

Soil community genomics or metagenomics is employed in this study to analyze the evolutionary related - ness of mangrove microbial community. The metagenomic DNA was isolated from mangrove sediment and 16SrDNA was amplified using universal primers. The amplicons were ligated into pTZ57R/T cloning vector and transformed onto E. coli JM109 host cells. The recombinant plasmids were isolated from positive clones and the insert was confirmed by its reamplification. The amplicons were subjected to Amplified Ribosomal DNA Restriction Analysis (ARDRA) using three different tetra cutter restriction enzymes namely Sau3A1, Hha1 and HpaII. The 16SrDNA insert were sequenced and their identity was determined. The sequences were submitted to NCBI database and accession numbers obtained. The phylo - genetic tree was constructed based on Neighbor-Joining technique. Clones belonged to two major phyla of the bacterial domain, namely Firmicutes and Proteobacteria, with members of Firmicutes predominating. The microbial diversity of the mangrove sediment was explored in this manner.

Novel esterases from microbes through classical and metagenomics approach: Studies on the enzymes and their applications

Cochin University of Science And Technology