Biotechnology

The Fluorescence (FL) and Anti Microbial (AM) effects of the LAlaninium maleate which is Zwitter ionic in nature is studied and reported, XRD data reveals that it is orthorhombic and space group as
P212121 and of NLO with SHG type. The FL emission wavelength of LAM is 399 nm for macro scale and 403 nm, 406 nm, 430 nm for nano scales of 50nm, 29nm and 27 nm of sample sizes. The Anti microbial analysis is also studied for macro scale and for nano scales of LAM for 4 species such as
Staphyloccus Sp., Pseudomonas sp., E.coli sp., Klebsiella sp., respectively.

The LVP macro crystals are grown by slow evaporations solution growth method and the empirical formula is C11H14N4O9, with a = 9.9715 Å, b = 6.2931 Å, c = 12.649 Å, β = 110.51°, crystal system as monoclinic with space group P21, the macro level NLO SHG value is 1.35 times than that of standard KDP which is predominant for opto electronic applications. Here its AD activity is analysed for macro and nano level samples and nano it is 18 nm.

The LVP macro crystals are grownup by slow evaporation solution growth method and the lattice parameters are a = 9.9715 Å, b = 6.2931 Å, c = 12.649 Å, b = 110.51
, crystal system is monoclinic with space group P21, the macro level NLO SHG value is higher than that of standard KDP which is predominant for opto electronic applications. Here its Anti microbial (AM) and fluorescence (FL) activities are analysed for macro and nano level samples and nano it is 18 nm is mainly taken for consideration and FL emission is of 376 nm, 399 nm, 413 nm, 425 nm for macro and 3 nano samples and AM is analysed for 4 species such as Staphyloccus sp., Pseudomonas sp., E. coli sp., Klebsiella sp., respectively.

A study on codon usage bias in cytochrome c oxidase I (COI) gene of solitary ascidian Herdmania momus Savigny, 1816.

Four ascidians, Didemnum psammathodes, Eudistoma muscosum, Lissoclinum bistratum and L. fragile extracts were used to evaluate antimitotic effects in root tip cells of Allium cepa. All the extracts caused an increase in mitotic index of A. cepa treated cells with increasing concentrations of treatment in all cases. However, roots treated with the highest concentration showed decreased in their mitotic index. The results of this study showed that, of the four analyzed ascidians, the methanol extract of L. bistratum had excellent antimitotic activity which is followed by D. psammathodes, L. fragile and E. muscosum with the Mitotic Index of 18, 24, 38 and 46 respectively. The number of cells entering prophase decreased with increase in concentration of the extracts. This finding suggests that the promising antimitotic properties of L. bistratum could be exploited in against cancer cells.

Ascidians are one of the dominant marine sedentary filter feeders recorded more frequently as introduced species than other taxa. It is renowned that artificial structures offer novel niches to the non-native species. A yearlong investigation was carried out to understand the role of ascidian colonization on various artificial structures located along 84 stations stretched on the 1076 km long Tamil Nadu coast of South India. It revealed the occurrence of 26 ascidian species, among these18 specimens were identified to species level, 8 were identified to genus level based on morphological characters. As on origin and nativity, out of the total 26, 3 species were classified as introduced, 8 species were classified as native and 15 as cryptogenic species. Interestingly, Polyclinum isipingense and Diplosoma variostigmatum were reported first time in Indian waters. The cryptogenic and colonial forms of ascidians are dominant in the artificial structures. There were significant differences observed between artificial structure type, geographic locations (p = 0.0071) and between ascidians forms as well as geographic areas (p = 0.00375). This study also confirms the artificial structures offer new niches for non-native ascidian colonization. The influence of the substrate (structure type) as well as geographic locations on the biotic assemblage was also observed.

DNA barcoding and phylogenetic analysis of five ascidians (Phlebobranchia) distributed in Gulf of Mannar, India.

Ascidians (Tunicates/Urochordates) are excellent animal models for developmental, evolutionary and molecular biology studies. Isolation of DNA is the basic phase for most of the molecular biology studies including DNA barcoding. So far there is no definite method for the isolation of DNA from ascidians. In an effort to overwhelm this problem, three different methods such as Phenol-Chloroform, Salting out and Kit method were evaluated for DNA extraction. Results showed that among the three methods, kit method, after few modifications, yielded sufficient quantity and quality of DNA. The yield of DNA between the samples and methods was significant (P<0.005). This method is more pronounced for the PCR amplification of COI gene with tunicate primer.

Traditional medicine is an important resource for the development of potentially useful new chemotherapeutic agents. The essential values and uses of some plants have been worked out and published, but many of them remain unexplored to date. In this study, acetone extract of leaves of Tridax procumbens was subjected to Phytochemical Screening, GC-MS study and Antioxidant activity. The results of the phytochemical analysis showed the presence of vital secondary metabolites in acetone extract which play a role in plant disease resistant mechanism. Among the 15 compounds obtained from GCMS study, the major constituents were Dodecane, 2,6,10-trimethly-, alkane compound with a tune of, Hexadecanol, Tridecanol, Fatty acid ester and Triterpenoid. This extract showed massive antioxidant activity. Inhibitory activity was increasing with increasing concentration with maximum 64.95% which indicates the presence of strong antioxidant activity. The range of inhibitory percentage of extract of T. procumbens was between 30 -65 with IC50 value of 200μg.

Use of microbes is very promising approach for the synthesis of technology based, ecofriendly, cost effective and biocompatible nanoparticles possessing unique physical and chemical properties is one of the developments in nanobiotechnology. Bacterial flora obtained from the tannery effluent was identified as Pseudomonas putida. Biosynthesis of silver nanoparticles (AgNPs) by using culture supernatant of P. putida was investigated and the formation of silver nanoparticles was confirmed by the change in colour of the culture filtrate from yellow to brown. Furthermore, the silver nanoparticles were characterized by means of UV-Visible spectroscopy and it showed absorption peak at around 440 nm which corresponds to the Plasmon resonance of silver nanoparticles. The XRD pattern showed the presence of sharp reflections at (111), (200), (220), (311) which indicate the presence of biologically synthesis of silver nanoparticles. The Scanning electron micrograph showed the spherical natures of particles have size ranged from 20-100 nm and possess an average size of 60 nm. A highly significant antimicrobial activity against pathogenic bacteria by the bio fabricated Ag NPs
was also revealed by the highest zone of inhibition of 20 mm dia. Therefore, this novel bacterial strain could be used for biofabrication of AgNPs and ultimately in the nanoparticle based drug formulation for the treatment of bacterial diseases.

The effect of free and immobilized cells of effluent tolerant bacteria isolated from the tannery effluents of Vaniyambadi to treat effluent water was studied. Two bacterial strains were isolated from tannery waste water and identified as Pseudomonas putida and Bacillus cereus. The level of turbidity and other Physico-chemical parameters found to decreases from high to moderate or optimum level in the effluent treated with free and immobilized cells of both isolated strains whereas, the pH was increased from 6.5 to 7.0. The percentage reduction of free CO2, total alkalinity, hardness,
dissolved oxygen, Nitrate and Nitrite was ranged from 45 to 75% when treated with P. putida, whereas they were ranged from 60 to 79% in the 15thday of the treatment. The level of BOD was reduced drastically after 15thday of exposure by the free cells of both bacteria. The maximum percentage of reduction (83%) was shown by immobilized B. cereus.The percent reduction of COD ranged from 16% to 65% in both the bacterial strains. The percentage of removal of Cr (VI) was 86 and 91% for free and immobilized cells respectively. In B. cereus treated sample, it reached 84% and 89% at the 15thday respectively. It is concluded that both P. putida and B. cereus investigated in this study are highly recommended for beneficial bioremediation applications for in-situ and off-site removal of pollutants.

In-vitro influence of mercury toxicity was tested on fresh water fish
Cirrhinusmrigala. The 96 h LC50 value in the acute toxicity test was 0.23 ppm for
mercury against the test fish. The results showed a decrease in biochemical constituents
with respect to increases in exposure periods. Activities of amylase, maltase and
invertase were suppressed to a minimum strength when exposed to Hg. When
experimental days progressed, all these enzymes showed only their presence and the
activities were found to be very minimum. Absence of protease and lipase enzymes
was observed in both control and experimental fishes. Among the 3 physiological
grouping of bacteria, lipolytic bacteria were found to be high followed by caseinolytic
and amylolytic bacteria. During accumulation periods, the number of these groups
reduced substantially with maximum 95% of amylolytic bacterial population in mid
gut which is followed by caseinolytic bacterial population.

DNA barcoding has played a significant role in biodiversity assessment as well as its conservation. This technique involves sequencing of mitochondrial marker gene including a short COI gene, known as barcode gene. It has proved its efficiency in identifying several species and resolving the limitations incurred during identification through conventional taxonomy. This study involves the use of DNA barcoding of ascidian species belonging to order Phlebobranchia. A total of 14 individuals, covering two families, three genera and five species, were barcoded. COI gene sequences of all the five species were deposited for the first time in NCBI as well as BOLD. The NJ tree revealed identical phylogenetic relationship among the individuals collected from three different stations. Mean Kimura 2-parameter (K2P) distances within-species, genus, family and order were 0.08%, 6.69%, 9.49% and 18.58%, respectively. This result concludes that COI gene sequencing is the efficient tool in identifying ascidians of the order Phlebobranchia. We report for the first time the COI gene sequences of four species of ascidians studied.

Ascidians (tunicates) are marine benthic organisms possessing various pharmacological activities, including anti-oxidant, anti-tumour, antimicrobial, etc. They also play a key role as model organisms to study various neurobehavioral disorders. Ascidian diversity is reportedly less in India due to lack of taxonomists as well as the limitations in morphology based taxonomy. Molecular taxonomy, comprising the sequencing of cytochrome c oxidase 1 gene (barcode region) otherwise known as DNA barcoding reduces these bottlenecks. Since several species of the family Didemnidae closely resemble in morphology, the present study was aimed to develop DNA barcodes of a colonial ascidian, Lissoclinum fragile belonging to the family Didemnidae. CO1 gene of L. fragile from Thoothukudi, Mandapam, and Vizhinjam waters were sequenced and submitted in GenBank, NCBI through Barcode submission tool. BLAST results showed maximum identity (97–100%) for L. fragile collected from different stations. The pairwise genetic distances within species and genera were calculated using Kimura two parameter (K2P) and the phylogenetic tree was constructed using Neighbour-Joining Tree.

DNA barcoding involving the sequencing of a short mitochondrial DNA segment, cytochrome c oxidase subunit 1 (COI) gene, is a specialized technique for the identification of species even at the early embryonic and larval stages, which is quite difficult in morphology-based taxonomy. Ascidians are sessile invertebrate chordates possessing numerous biochemical as well as pharmacological activities. In this study, a total of 36 ascidian samples belonging to the family Didemnidae were sequenced for a 650 bp region of the mitochondrial COI gene. All the species were represented by multiple specimens. The barcode sequences showed no stop-codons and indels in the alignments. The aligned sequences were submitted in Barcode submission tool, NCBI, and the accession numbers were obtained. The minimum intraspecific distance was found to be 0.00% and the maximum was 2.23%. Mean Kimura 2-parameter (K2P) distances within-species, genus, and family were 0.88, 5.98, and 20.03%, respectively. The mean interspecific distance is more than the mean intraspecific divergence, which indicates efficiency of DNA barcoding for identification of ascidians.

Ascidians are well known worldwide for their rapid invasions and also for the presence of potential biomedical molecules. Members of the family Didemnidae are widely distributed in tropical waters and they are reported to be among the families possessing rich bioactive compounds. Didemnum psammathodes has a cosmopolitan distribution in tropical waters. The growing evidence of multifarious potential and ever increasing invasion of this species accentuated the need for additional research into its diversity and distribution for sustainable utilization and conservation. The present study was intended to focus on distribution and invasiveness of colonial ascidian, D. psammathodes, along the southern Indian peninsular waters. The present data are based on our own observations made during 2012—2014 period and also on the published and unpublished records of the last 20 years. Out of 45 stations surveyed, D. psammathodes was encountered at a maximum of 41 stations and was found to be more abundant in Hare Island (n = 42), North Break Water (n = 38) and Vizhinjam bay (n = 32). This species was absent at four different stations. Catch per unit effort was higher (19.6) in Hare Island followed by NBW (16.0) and Vizhinjam bay (6.8). The highest number of colonies (136) was observed in calcareous stones, followed by embedded rocks (54) and molluscan shells (33). Hydrographical parameters showed no significant differences between the stations ( p < 0.005). It is concluded that D. psammathodes has the potential to invade most of the stations and its distribution was not influenced by hydrographical parameters rather than substrates.

Background
Ascidians play a key role in the ecology and biodiversity of marine ecosystem. Ascidians
can be transported in ship ballast water and while attached to ship and boat hulls. Heavy
traffic by domestic and international ships as well as cargo vessels between the major and
minor ports warrants continuous monitoring for new introductions of ascidians. The
Mandapam coast is situated in the Gulf of Mannar, India, a marine hot spot area in the
Indian Ocean which provides an environment suitable for the settlement of ascidians.
New information
A total of 30 species of ascidians were reported from Mandapam coastal waters, of which
26 species were new to the study area and five species: Ecteinascidia turbinata, Eudistoma
carnosum, Trididemnum caelatum, T. vermiforme and Didemnum spadix, were new to
India.

The giant freshwater prawn Macrobrachium rosenbergii is the largest and fastest growing species and considered to be a potential candidate species for large scale aquaculture because of its capacity to breed in captivity, availability of well established techniques for larval rearing, a very good growth rate, production and survival and wide consumer acceptability. Further, the absence of major disease problems is also favourable for farming this species. However, suitable technique
to raise broodstock in this species is scanty. In the present study, two hormones namely Human chorionic gonadotropin and Diethylstilbestrol were selected to raise growth and development in short period. The two selected hormones were incorporated separately in a protein rich (32%) pelleted feed at l ppm, 1.5 ppm and 2 ppm concentrations. M. rosenbergii were fed with these hormone incorporated feeds to study the growth of the animal. In the first experiment, the juveniles were
fed with the hormone incorporated feed to study the growth and maturation. The experiment was conducted for 120 days. Of the different concentrations tested for the two selected hormones, growth was found to maximum for animals fed with hormone HCG at 1.5ppm concentration. In the second experiment, maturing M.rosenbergii were reared and fed with the two selected hormones namely HCG and DES at three concentrations (l ppm, 1.5ppm. 2ppm). The experiment was conducted for 90 days. M. rosenbergii fed with feed HCG incorporated feed at 1.5ppm concentration attained good growth (35g for male) and (37.15g for female). In both the experiments, the monthly growth rate was found to be high in animals fed with HCG incorporated feed at 1.5ppm concentration. The obtained findings revealed that M. rosenbergii fed the diet containing HCG is recommended to obtain
optimum growth performance and feed utilization.

Indian coastline is dotted with 12 major ports and number of minor ports and provided with diverse habitats which are ideal for the settlement of number of ascidians. Even though India has very long coastline extending to about 7000 km, only limited species of ascidian have been recorded. Exhaust review of literature revealed that there are untouched and undisturbed areas available in Indian regimes. These areas are to be explored not only for conserving the biodiversity but also for monitoring the alarming entry of exotic species through ballast water and hull of ships, as these species have significant effects on biodiversity rich area. In this context, the present study is aimed to report ascidians of two ecologicallysignificant stations such as Hare Island and Inigo Nagar located
along the Thoothukudi coast. A total 20 species of ascidians have been reported. Station 1 was found to harbour seven species of ascidians which are exclusively colonial whereas 15 species of ascidians with only two simple ascidians has been found at station 2. Among 15 species, Microcosmus curuvus and Styela canopus were predominant whereas at station 1, Eudistoma viride and E. muscosum were dominant. As this preliminary work could give as many as 20 species of ascidians in only two untapped areas, a detailed survey on various unexplored areas along with seasonal availability, factors affecting colonization, succession at different depths etc is sure to yield a rich haul of ascidians in future.

Occurrence of alien ascidians in VO Chidambaranar port, Thoothukudi

Morphology-based taxonomical studies of ascidians in India are meagre due to lack of ascidian
taxonomist and limitations inherent in conventional system-based identification. The use of
short fragment of mitochondrial DNA sequence is proving highly useful in identifying species in
a situation where, the traditional morphology-based identification is difficult. In the present
study, two adult solitary ascidians collected from the Thoothukudi coast were morphologically
identified as Herdmania momus Savigny, 1816 and Microcosmus squamiger Michaelsen, 1927.
The genomic DNA of these ascidians was isolated, COI gene was amplified, sequenced and
submitted to the GenBank under the accession numbers KM058116, KM411616 and KJ944390.
Homology search result using BLAST showed that H. momus showed 100% matched with other
H. momus, while M. squamiger showed similarity with Pyura herdmani, a member of the same
family Pyuridae. The phylogenetic and genetic distance was maximum in interspecies than in
intraspecies. These COI sequences will allow the identification of the species through DNA
barcoding technique. Here, we report for the first time the COI gene of H. momus, Savigny 1816
from the Indian coast.

Globally, ascidians is proven to have significant novel lead molecules with varying bioactivities, but have remained poorly studied in this context in India. In vitro antibacterial screening of a colonial ascidian Lissoclinum bistratum collected from Hare Island, Thoothukudi coast of India, against selected clinical isolates of bacteria was carried out in this study. The methanol extracts of L. bistratum exhibited broad antibacterial activity. The highest activity was shown against Pseudomonas aeruginosa (inhibitory zone of 11mm in DIA).GC-MS study revealed the presence of 14 chemical
constituents. Of these different compounds, Phenol, 2,5-bis (1,1-dimethylethyl), Benzoic acid, 4-ethoxy- ethyl ester, Dodecanoicacid, Acetic acid, octadecyl ester, Oleyl Alcohol, 1-Eicosanol, Ethanol, and 2 (octadecyloxy)- are known for antimicrobial activity. It is concluded that there is a great scope for finding further novel antimicrobial compounds in the ascidian.

Accurate and unambiguous identification of marine animals, from eggs to adult, is important for conservation of biodiversity. Conventional morphological identification of species has many limitations particularly in larval and egg stages. This can be addressed through various protein and DNA based methods of genetic identification. Ascidians are sessile and filter-feeding marine animals and are
known for several pharmacologically important metabolites. Species identification of ascidians is possible by observing various morphological and anatomical features in various stages of life span. However this method is labour intensive, time consuming and very difficult for non-specialists particularly when dealing with field collections. This study is aimed to identify morphologically similar
colonial ascidians collected from Mandapam coast through molecular technique i.e. DNA barcoding. For the present study, the whole genomic DNA of both species 1 and 2 was isolated using Qiagen kit and the cytochrome c oxidase sub unit 1 gene (CO1) of each species was amplified with Universal Primers. Forward and reverse sequences of Species 1 contains 910 and 589 bp respectively whereas Species 2 contains 708 and 629 bp respectively. BLASTn result of both forward and reverse sequences of the Species 1 showed 99 and 98% identity with the species Didemnum perlucidum (JQ731735.1 and JQ731740.1). Similarly BLASTn result of the Species 2 showed 99 and 98% identity with the species Lissoclinum fragile (KJ725150.1, KM411612.1 and KM411611.1). This result
confirmed the identification of species 1 as Didemnum perlucidum and species 2 as Lissoclinum fragile. This study clearly shows that CO1 gene sequence will help the identification of ascidians through DNA barcoding.

In-silico analysis of CO1 gene sequence of two colonial ascidians Polyclinum indicum and Didemnum candidum from Gulf of Mannar

Ascidians (Tunicata) are gaining paramount importance in raising marine biodiversity. Considering ongoing changes in marine ecosystem, seasonality in the recruitment of ascidians and their invasive nature, the arrival of ascidians is increasing in India. In this context, a field study was carried out in selected transects of South Indian coastline during 2012–2014 to understand occurrence and distribution of ascidians. The study revealed the occurrence of 36 species of ascidians belonging to 7 families and 17 genera inclusive of 2 new records. The highest number of species was found in the
family Didemnidae followed by Styelidae. The most abundant colonial species were Eudistoma pyriforme, E. microlarvum, Botrylloides magnicoecum, B. schlosseri, Phallusia nigra, Aplidium multiplicatum, Didemnum psammathodes, Lissoclinum fragile, L. bistratumandEudistomasluiteri. Botrylloidesmagnicoecum, B.leachie and B. chevelence were present exclusively in Colachel. Among the 37species, D. psammathodes and L. fragile were found majority of the stations studied. E. microlarvum and E. sluiteri were reported for the first time at Tiruchendur water. Four species of the sub-family Botryllinae of Styelidae and one species of Didemnids were fouling the entire hull of boat at Colachel station. An increasing trend in the occurrence of ascidians in southern Indian waters reinforces the need for continued and periodic monitoring of their population expansion and also for sustainable utilization towards human welfare.

Ascidians belonging to the sub-phylum Uro-chordata are used as potential model organisms in various parts of the world for biosorption of metals.The sedentary nature, filter feeding habits, presence of vanadocytes and the absence of kidneys cause them to accumulate metals.The present study was aimed to compare biosorption of metals such as cadmium,copper,lead,mercury and vanadium in test and mantle body of solitary ascidian Phallusia nigra between two ecologically significant stations such as Thoothukudi (Station1) and Vizhinjam (Station2) ports of India.Monthly Sampling Of Water And P.nigra were done for a period of one year from September 2010 to August 2011 and subjected to analysis of metal accumulation.The average metal concentrations except mercury in the Thoothukudi water were found to be higher of comparable magnitudes than the Vizhinjam water.One-way ANOVA showed significant differences between the stations.A comparison of average metal concentrations in the test and mantle bodyof P.nigra between two stations showed that the enrichment of V, Cd, Pb, Cu and Hg in the Thoothukudi samples may be due to high bioaccumulation factors of these elements as compared to other species of ascidians.The bioaccumulation factors were in the order of V> Pb >Cd >Cu > Hg for the test and mantle body in stations 1 and 2. Application of one- way ANOVA for the concentration of these metals between test and mantle body showed significant differences in both stations.Similarly, ANOVA for biosorption of these trace metals by P.nigra showed significant differences between stations. Metal concentrations recorded in this ascidian could effectively be used as good reference material for monitoring metal contamination in Indian sea waters.

The mitochondrial cytochrome c oxidase subunit I (COI) gene, a widely accepted molecular
marker for species identification and classification, has been questioned because of the
presence of Numts. In this study we found the presence of Numts in the COI chromatogram
of two tunicates, Polyclinum indicum and Polyclinum madrasensis belonging to the genus
Polyclinum. Numts were also present in our sequence (Accession Number: KJ944391) and in
other sequences belonging to genus Polyclinum in the GenBank record. The GeneBank
database of genus Polyclinum contains COI-like sequences and COI pseudogenes, but no record
of COI gene from Polyclinids. The prevalence of Numts in Polyclinids belonging to Tunicata,
is an impediment to DNA barcoding studies of Polyclinum species.

More recently, ascidians belonging to the sub-phylum Uro-chordata are used as potential model organisms in various parts of the world for biosorption of metals. In India, the study on accumulation of metals in ascidians is meagre. Hence the present study is aimed to analyze the concentration of metals in selected five species of ascidians and their environment in Thoothukudi coast, India. Among the four metals studied in seawater, Cu accumulated in highest concentration (0.0028
ppm) followed by Pb (0.0026 ppm), V (0.0023 ppm) and Cd (0.0021 ppm). Biosorption of four metals was in the order of Cu>V>Pb>Cd in Microcosmus squamiger, Microcosmus exasperates and Herdmania pallida. But, in Phallusia arabica and Styela canopus, the order was V>Cu>Pb>Cd. The concentration factors were in the following order Cu >V >Pb> Cd in all ascidians studied except
in P. arabica where, it was V > Cu>Pb>Cdand BAF was 9665, 2176, 592 and 7.1 for V, Cu, Pb and Cd respectively. The BAF is high in P.arabica followed by H. pallida, S. canopus, M. squamiger and M. exasperates. Thus, it is concluded that ascidians could be used as bioindicator of metals in seawater.

Scientific facts in the Tamil proverb on cow dung.

Synthesis and antimicrobial activity of salicyloyl chitosan and starch blend.

Development of biogenic silver nano particle from Pelargonium graveolens leaf extract and their antibacterial activity

Diversity and seasonal variations of Class Ascidiacea in Thoothukudi coast, India

The present investigation was aimed to evaluate efficacy of ethanolic extract seeds of
Nigella sativa against glucose induced cataract in goat eye lens. In the in vitro study, goat lenses were
subjected to photographic evaluation and biochemical parameters such as protein, GSH, MDA,
NA+/K+ ATPase and sodium and potassium were also analyzed. Photographic examination of the eyes
showed that treatment with ethanolic extracts of seeds of N. sativa retarded the progression of lens
opacification. Cataract lens treated with N. sativa elevated the activity of Na+ K+ ATPase, total and
water soluble proteins and K+ ions to the level of normal level whereas reduced concentrations of Na+
ions. The MDA levels were significantly less in the N. sativa treated groups whereas, the level of
GSH, in high glucose (55mM), compared to the normal control group was significantly low but N.
sativa treated groups showed higher level of GSH. These results support the view that ethanolic
extract of seeds of N. sativa as seen in this in vitro model may, counteracts the effects of glucose in
inducing cataract to some extent.

New record of colonial ascidians from south west coast off Indi

Nutritional value of solitary ascidian Microcosmus exasperates Heller, 1878 (Ascididae: Pyuridae)

Herdmania pallida, simple ascidian belongs to the Subphylum Urochordata, Class Ascidiacea
commonly available in Tuticorin coast. Like other animals such as prawns, molluscs and fishes, ascidians are also consumed as food in various parts of the world due to its high nutritional and medicinal value. In the present study, biochemical components like carbohydrate, protein, lipids and amino acids were quantitatively estimated in H. pallida. As this species has high protein content, in its body tissues, pickle was prepared from the mantle bodies and the product was packed in sterilized bottles and stored at ambient temperature. The shelf life period was assessed for a period of six months. Quality indicators like pH, microbial load and sensory evaluation were analysed. Spoilage was not observed through out the study period. In conclusion, the prepared pickle from the mantle bodies of H. pallida is good and safe product for human consumption and can be stored for a period of six months.

The Indian peninsular is dotted with 12 major ports and a number of minor ports serving as gateway
for non-indigenous marine organisms. Taxonomical studies on ascidians in India have been expanded but there is a lack of information about the distributions of alien ascidians. Hence the present study was aimed to know the distribution of alien ascidians at 11 stations in the east and west coasts of Indian peninsular. A maximum of 34 non-indigenous ascidians out of 41 species have been reported in the present study. They all belong to diverse families with maximum representation from Didemnidae. All eight native ascidian species are exclusively colonial. Among the 34 alien ascidians, 26 species are listed as cryptogenic of which two are established-cryptogenic (Herdmania pallida and Lissoclinum fragile). Eight species are invasive with two established-invasive species such as Phallusia nigra and Didemnum candidum. The highest ascidian diversity was noticed in Tuticorin (south east coast) and Kadiapattanam (south west coast).

In the present observation, the food and feeding habit of the simple ascidians, Herdmania pallida
was carried out during the study period (July 2008 to June 2009). The percentage contribution of food items comprised of 51% of sand particles, 30% of micro flora, 13 % of meio fauna, 2% of macro fauna and 4% of other materials. The percentage of feeding intensity were observed throughout one year as poorly feed 20%, heavy feed 26%, medium 18 %, active 24% and empty 12%, The poorly feed were more recorded in the month of June (32.5%), heavy feed was highly recorded in the month of January (37.5 %), active feed was more reported in the month of February (34.6%),medium feed was more in the month of January (20.2%), empty feed was reported high in the month of July (18.9%).

The Indian peninsular is dotted with 12 major ports and a number of minor ports serving as gateway
for non-indigenous marine organisms. Taxonomical studies on ascidians in India have been expanded but there is a lack of information about the distributions of alien ascidians. Hence the present study was aimed to know the distribution of alien ascidians at 11 stations in the east and west coasts of Indian peninsular. A maximum of 34 non-indigenous ascidians out of 41 species have been reported in the present study. They all belong to diverse families with maximum representation from Didemnidae. All eight native ascidian species are exclusively colonial. Among the 34 alien ascidians, 26 species are listed as cryptogenic of which two are established-cryptogenic (Herdmania pallida and Lissoclinum fragile). Eight species are invasive with two established-invasive species such as Phallusia nigra and Didemnum candidum. The highest ascidian diversity was noticed in Tuticorin (south east coast) and Kadiapattanam (south west coast).

This study was carried out in order to screen the antibacterial activity of the bioactive compounds
extracted from the test body (body covering) and the mantle body of the two solitary ascidians,
Phallusia nigra and Herdmania pallida, collected from the Tuticorin harbour area. Antibacterial
activity of P. nigra was significantly higher than that displayed by H. pallida. The former species
showed maximum antibacterial activity and inhibition against all the bacteria tested. The bacterial
growth inhibitory effect was found to be dependant on the solvents used for extraction as
well as on the specific to the species extracts. Of the nine strains examined, Gram positive were
the most susceptible after treatment with all fractions except fraction VI which showed activity
against Escherichia coli and Salmonella paratyphii (Gram negative). Lipophilic (chloroform) extracts
were highly active against the pathogens tested. The study was also extended to find out
the chemical nature of the active compound. IR, 1H NMR and 13C NMR spectra of the fraction
II provided the complete carbon skeleton of a tyrosine derivative. This tyrosine derivative could
be probably responsible for the antibacterial activity and a future study involving pharmacology
would pave a way for the invention of a new antibiotic drug from P. nigra.

The aim of this study was to assess the effect of phosalone on the tilapia, Oreochromis mossambicus. The effect was assessed on the basis of the results of acute toxicity tests and comparison of results of haematological examination of a control and an experimental group exposed to phosalone. The 96 h LC50 is the basic value in the acute toxicity test and it was 0.26 mg/L for phosalone against the test fish. Haematological profile of the healthy fish was carried out with 2 sublethal concentrations, viz 0.0325 mg/L (1/8) and 0.065 mg/L (1/4) of phosalone. The experiment was performed semi-statically with a group of 10 fish in 10 experimental aquaria, 2 control aquaria, and 4 aquaria each with different concentrations. The results showed that phosalone had some effect on the haematological parameters of tilapia. It was determined that the total RBCs, WBCs, haemoglobin content, and haematocrit value significantly decreased (P < 0.001). The percentages of erythrocyte sedimentation rate, mean corpuscular haemoglobin, and mean corpuscular haemoglobin
concentration showed an increasing trend with respect to the increase in exposure period in both sublethal concentrations. ANOVA showed that the pesticide concentrations had more influence than the duration of exposure in all cases (P < 0.001).

Occurrence and distribution of ascidians in Vizhinjam Bay (south west coast of India)

DNA barcoding, which involves use of a short fragment ̴ 650 bps of mitochondrial cytochrome C oxidase subunit 1 (COI) gene as a
marker for the identification of species, has been proposed as a global standard for identifying unknown biological material to species level.
Since morphology based identification of ascidians has many limitations, there is need for molecular level identification for accurate
identification at species level. Three different habitats of Vizhinjam Bay were sampled for ascidians during the study period from May 2016 to
July 2017. COI gene sequences of 27 samples were amplified and the sequence data deposited in GenBank, NCBI. All submitted sequences
were released to public data base with accession numbers to all the sequences. As a result, a total of 17 species inclusive of 3 new records
(Diplosoma sp. BK02, Cnemidocarpa sp. BK03 and Diplosoma multifidum) under 10 genera and 5 families were recorded. The first report of
entry of three species in Indian coast was reported and CO1 gene sequences of these species were deposited in GenBank, NCBI. Ambiguity in
identification of Polyclinumsaturnium was confirmed with homology search in BLAST as Polyclinum indicum. Phylogenetic analysis through
NJ tree using K2P presents the branching pattern of the evolutionary relationship of the ascidians. Species that are most similar grouped
together to form a cluster. Clusters of species in a clade indicate that the species are closely related. Species that are highly divergent form a
separate branch.

Diversity and distribution of 42 species of ascidians belonging to 7 families and 19 genera from six different stations along the southwest coast of India were documented. Most of the species were recorded for the first time from the southwest coast of India while three species appear to be new records from India. Previous records of these species in India were usually from Gulf of Mannar particularly in Thoothukudi water. The family Didemnidae was represented by 13 species of 4 genera followed by Styelidae (11 species of 7 genera), Polyclinidae (7 species of 2 genera), Pyuridae (6 species of 2 genera), Perophoridae (two species) and Polycitoridae and Ascidiidae (one species each). The present study showed that the diversity and distribution of ascidians have changed considerably when compared to a previous report. Recruitment of ascidians in Vizhinjam Bay
region was more than in the other regions. As many as 28 species of ascidians were recorded in Vizhinjam Bay, 16 species in Colachel, six in Kadiapattanam, five in Chinna Muttom and
four each in Muttom and Leepuram. The percentage composition of representatives of various genera was: Didemnum and Polyclinum each by 15%, Microcosmus-12%, Botryllus- 8%,Trididemnum-10%, Botrylloides, Symplegma, Eudistoma and Diplosoma each by 5%; and Phallusia, Herdmania, Styela, Monandrocarpa, Polycarpa, Cnemidocarpa, Aplidium,
Lissoclinum, Perophora and Ecteinascidia each by 2%. The results showed that distribution of colonial ascidians were more at majority of the stations whereas the simple ascidians were
restricted to port area. Diversity, distribution and invasive status of ascidians are discussed in detail.

Non-indigenous ascidians in V.O.Chidambaram port, Thoothukudi, India.

Exploration of untapped resource of ascidians along the Thoothukudi coast of India.

Biosorption of metals by solitary ascidian, Phallusia nigra in Vizhinjam Bay (south west coast of India).

Marine ascidian biodiversity – a promising resource for bioactive compounds.

Ascidians – non conventional resources of food from marine environment

Marine tunicates collected from Tuticorin, southeast coast of India, were found to be a promising source of novel bioactive compounds against human urinary tract
pathogens. Methanol and ethyl acetate extracts of Phallusia nigra was tested against five human urinary tract pathogens. Streptomycin (400μg/ml) was used as a
standard and dimethyl formamide as control. Notably, both methanol and ethyl acetate extract of P. nigra showed activity against all the UTIs strains tested. The
methanol extract displayed higher antibacterial activity than ethyl acetate extract. Antibacterial activity of ascidians extracts increased with increasing concentrations.

Seasonal variation of ascidians (Tunicates) in Tuticorin coastal area

Biodiversity – the future of nature

Microbiology Laboratory Manual

Apiculture

Ascidians in Coastal Water: A Comprehensive Inventory of Ascidian Fauna of Southern Indian Coasts

DNA Barcoding: Methods and Protocols

Uyirinap Pazhamozhiyil Ariviyal Unmai

In India, 11 species of shrimp have been found to be suitable for culture among 23 species
available. Litopenaeus vannamei, Penaeus monodonand Macrobrachium rosenbergii are the major three
species contributing great economy in Indian aquaculture industry.Total freshwater aquaculture production
increased in 2016 compared to previous year.Aquaculture production of M. rosenbergii increased from 8,729
tonnes in 2015 to 10,152 tonnes in 2016. Giant freshwater prawn M. rosenbergii is becoming an important
targeted species, as its culture is considered to be the potential source of income among farmers. This study
reviews the current production status of freshwater prawn, white tail diseaseand its diagnostic methods and
potential drugs available to treat the prawn diseases.

Melia azedarach (Kattu vembhu - Tamil) also called as Persian lilac.In the present study, the phytochemical analysis showed that theMelia azedarach contains Alkaloids, Carbohydrate, Tannins, Saponin, Terpenoids, Oxalate and Glycoside and the absent of Amino Acid, Phenol, Flavonoids, Quinones and Coumarin in the Acetone extract. The Benzene extracts of Melia azedarach contains Alkaloids, Carbohydrate, Phenol, Saponin, Terpenoids, Coumarin and Glycoside and absence of Amino Acid, Flavonoids, Tannins, Oxalate and Quinones.Alkaloids, Carbohydrate, Tannins, Saponins, Terpenoids and Oxalate are presentand Amino Acid, Phenol, Flavonoids, Quinones, Coumarin, Glycosides are absent in the Methanol extract.Alkaloids, Amino Acid, Phenol, Flavonoids, Terpenoids, Quinones are presentand Carbohydrates, Tannins, Saponin, Oxalate, Coumarin and Glycosides are absent in the Water extract of Melia azedarach. Anthocyanin is absent in all the solvent extracts of Melia azedarach.Acetone, Benzene, Methanol and Water extract of Melia azedarach show activity against the tested pathogens by the highest zone of inhibition Staphylococcus aureus (10 mm), Escherichia coli (9 mm), Pseudomonas aeruginosa (7 mm) and Bacillus subtilis (5 mm)for Acetone Extract of Melia azedarach, Staphylococcus aureus (6 mm), Bacillus subtilis (5 mm), Escherichia coli (5 mm) and Pseudomonas aeruginosa (4 mm)recorded. Benzene Extract of Melia azedarach resulted inStaphylococcus aureus (16 mm), Escherichia coli (13 mm), Pseudomonas aeruginosa (9 mm) and Bacillus subtilis (6 mm) and for Methanol Extract of Melia azedarach and Pseudomonas aeruginosa (6 mm) and other pathogens are resistant to the Water extract of Melia azedarach.The antioxidant activity level of Melia azedarach solvent extracts was high to Benzene followed by Acetone, Methanol and Water. The results of the study indicate that the Melia azedarach possesses phyto-constituents having antibacterial activity thus it can be utilized as a natural plant based antimicrobials.

Isolate Pseudomoans fluoresces for studying the multiple antagonistic activity. The five different rhizosphere soil samples were collected and inoculated into selective media and their morphology was observed. Human skin swabs were collected for the isolation of five different pathogens such as Staphylococcus aureus, Streptococcus, Bacillus subtilis, Salmonella, Clostridium. The morphology characteristic of human skin pathogens causing microorganisms identified by performing Gram’s staining, capsule staining, motility test. The biochemical characteristics of five bacterial human pathogens causing micro organisms were studied. Pseudomonas fluorescens skin infections caused by various pathogens can be prevented.

Medicinal plants an important role in the healthcare of people around the world, especially in developing countries. About 80% of the population of most developing countries still use traditional medicines derived from plants for treating human diseases. The present study exposed that the people of the snake charmer community used 57 medicinal plant species that belonged to 51 genera and 35 families for the treatment of various diseases. The study has brought to light that the main diseases treated by this community was snakebite in which 19 different types of medicinal plants belongs to 13 families were used. Significantly higher number of medicinal plants was claimed by men as compared to women. The highest per cent of medicinal plants for traditional uses utilized by this community were belonging to family Fabaceae.

Ethnobotanical inspection was conducted on the Kalvarayan hills in the Vilupuram district, Tamil Nadu, India. The principle of this research is to collect data on medicinal plants available in the Kalvarai hills to preserve its rich biological resources through registries, as there is little information about the existence of medicinal plants in this region. This survey was conducted on the basis of semi-structured interviews, field visits and joint observations. A total of 20 medicinal plants were found and confirmed in the Herbarium of the University of Madras in Chennai. Plant species belonging to various families, such as Lilaceae, Solanaceae, Euphorbiaceae, followed by Lamiaceae and Apocyanaceae, have been found. These plants have medicinal properties, including anti-cancer, anti-diabetic, antidotes, antibacterial, anthelmintic, antispasmodic, anti-analgesics, healing of epilepsy, diarrhea, weakness, etc. This may be due to the growth of parthenic herbicide species. The presence of this herbicide led to a decrease in some medicinal plants in this area. Further research is needed to the hills slowly declining biological resources in this region.

An ethno botanical survey was conducted in Yelagiri hills, located in Vellore district, Tamil Nadu (State), India. The purpose of this study is to collect data about medicinal plants available in yelagiri hills in order to preserve its rich bio resources through documentation since there is scarce information about the existence of medicinal plants in this region. This survey was conducted based on semi structured interviews, field visits and participatory observations. A total of 28 medicinal plants were identified and confirmed in herbarium of university of madras, Chennai. Plant species belonging to diverse families like Lilaceae, Solanaceae, Euphorbiaceae followed by Lamiaceae, apocyanaceae were found. These plants have medicinal properties including anticancer, antidiabetic, antidote, antibacterial, anthelminthic, antispasmodic, antianalgesic, cure for epilepsy, diarrhea, weaknesses etc. However, certain medicinal plants were found to be in vulnerable condition like Ocimum sanctum and Leucas aspera. This could be due to growth of herbicide parthenium species. The presence of this herbicide has led to the decline of certain medicinal plants in this area. Further investigation is required to preserve the bioresources that is slowly declining in this region

The Fabaceae, the third largest family of plants and the source of many crops, has been the target of many genomic studies. Currently, only the grasses surpass the Chena legumes for the number of publicly available expressed sequence tags (ESTs). The quantity of sequences from diverse plants enables the use of computational approaches to identify novel genes in specific taxa. We used BLAST algorithms to compare unigene sets from Chickpea to nonlegume unigene sets, to GenBank’s nonredundant and EST databases, and to the genomic sequences of rice and Arabidopsis. As a working definition, putatively legume-specific genes had no sequence homology, below a specified threshold, to publicly available sequences of nonlegumes. As a first step toward predicting function, related sequences were clustered to build motifs that could be searched against protein databases. Evolutionary analyses of the genomic sequences of several CCPs in M. truncatula suggest that this family has evolved by local duplications and divergent selection.

250 objectives question and answers

In order to document the medicinal plants and their applications, a field survey
was undertaken at Mahadeva hill which is located 28 Km away from Vellore city
and 60 k.m. from Islamiah College (Autonomous), Vaniyambadi, TamilNadu,
India. An attempt has been made to identify the medicinal plants traditionally used
by the people of the region and also to preserve the traditional heritage. Further
this study highlights the risks of extensive human exploitation of biodiversity of
the region. The survey was carried out during January to November 2019. A total
of 88 herbaceous medicinal plants were documented and an inventory was
prepared. All these 88 plants species are belonging to 40 families such as
Acanthaceae, Aizoaceae, Alangiaceae, Alliaceae, Amaranthaceae, Annonaceae,
Apocyanaceae, Aristolochiaceae, Asclepiadaceae, Asteraceae, Bombacaceae,
Caesalpinacea, Capparaceae, Caricaceae, Cesalpinioideae, Cleomaceae,
Cucurbitaceae, Euphorbiaceae, Fabaceae, Gentianaceae, Labiatae, Lamiaceae,
Liliaceae, Lythraceae, Malvaceae, Meliaceae, Mimosaceae, Moraceae, Myrtaceae,
Oleaceae, Oxalidaceae, Papilionoideae, Poaceae, Rhamnaceae, Rubiaceae,
Rutaceae, Sapindaceae, Solanaceae, Verbanaceae and Vitaceae.This quantitative
survey gives an account on the floral diversity and uses of medicinal plants.

The phytochemical analysis of the various solvents extract such as Chloroform,
Hexane, Propanol and Ether were tested for 14 phytochemicals like Alkaloids,
Flavonoids, Saponin, Terpenoids, Glycoside, Amino Acid, Phenol, Carbohydrate,
Tannins, Quinones, Phlabotanins, Anthocyanin, Oxalate and Coumarin for Senna
alexandrina and Coccinia grandis. The zone of inhibition for Senna alexandrina
solvent extracts for Chloroform (8 mm), Hexane (8 mm), Petroleum Ether (5
mm), Propanol (5 mm) for Staphylococcus aureus bacterium and Propanol (8 mm),
Petroleum Ether (8 mm), Hexane (6 mm), Chloroform (4 mm) for Pseudomonas
aeruginosa bacterium are shown. The antibacterial activity of Coccinia grandis
for Propanol (16 mm), Petroleum Ether (10 mm), Chloroform (8 mm), Hexane (7
mm) for Staphylococcus aureus and Propanol (13 mm), Petroleum Ether (6 mm),
Chloroform (6 mm), Hexane (5 mm) for Pseudomonas aeruginosa are shown. The
results of the study indicate that both Senna alexandrina and Coccinia grandis
possesses phyto-constituents having antibacterial activity thus it can be utilized as
a natural plant based antimicrobials.

Ocimum sanctum has a rich and fanciful history known since the Vedic age for its immense curative and multi-purpose utility. The phytochemical results show the presence of Amino Acid, Carbohydrate, Phenol and Terpenoids in all the four solvent extracts of the O. santum. Except Aqueous extract of the plant all other extracts show the positive results for alkaloids, Coumarin, Flavonoids and Glycoside. Saponin is present in the benzene and water solvent extracts of O. sanctum. The four solvent extracts are showing the absence of following phytochemical�s such as Tannins, Phlobatanins, Quinones and Anthocyanin. The higher concentration Acetone extract, Benzene extract, Methanol extract and Water extract of the O. sanctum show the activity against the tested pathogens by the following order from the highest zone of inhibition P. aeruginosa (8.5 mm), S. typhi (5 mm) and E. coli (3.5 mm) for Acetone extract, E. coli (4 mm) only for Benzene extract, whereas P. aeruginosa (14 mm), S. typhi (8 mm) and E. coli (7 mm) for Methanol extract and there is no activity for the water extract for all the three pathogens were observed. The antioxidant activity of the O. sanctum solvent extracts was high to Methanol followed by Benzene, Acetone, Water extracts. The results of the study indicate that the Ocimum sanctum possesses phyto-constituents having antibacterial activity thus it can be utilized as a natural plant based antimicrobials.

Azolla pinnata is an aquatic nitrogen fixing pteridophyte commonly found in aquatic habitat. In the present study, the phytochemical analysis showed thatAlkaloids, phenol, terpenoids and oxalate are present in four solvent extracts except water. Carbohydrate is absent in benzene but present in all other extracts. Flavonoids present in acetone and water extract, saponin is present in methanol and water extract.Ethanol and water extract exhibited the presence of anthocyanin, except benzene and methanol other extracts contain Coumarin. The phytochemicals tannins, Phlobatanins and glycosides are absent in all the solvent extracts of A. pinnata. This study was carried out against the various extracts namely Acetone, Benzene, Ethanol, Methanol and Water of Azolla pinnata for its antibacterial activity against Pseudomonas aeruginosa and Staphylococcus aureus. The zone of inhibition S. areus (13 mm), P. aeruginosa (10 mm)for Acetone extract, S. areus (12 mm), P. aeruginosa (8 mm) for Benzene Extract, P. aeruginosa (12 mm), S. areus (9 mm) for Ethanol extract, S. areus (12 mm), P. aeruginosa (11 mm) for Methanol extract are observed and there is no activity for the Water extract of A. pinnata.The antioxidant activity of A. pinnata solvents extracts was more to the Ethanol followed by Methanol, Acetone, Benzene and Water sample. The results of the study indicate that the Azolla pinnata possesses phyto-constituents having antibacterial activity and thus if can be utilized as a natural plant based antimicrobial.

Abstract

Melia azedarach (Kattu vembhu - Tamil) also called as Persian lilac.In the present study, the phytochemical analysis showed that theMelia azedarach contains Alkaloids, Carbohydrate, Tannins, Saponin, Terpenoids, Oxalate and Glycoside and the absent of Amino Acid, Phenol, Flavonoids, Quinones and Coumarin in the Acetone extract. The Benzene extracts of Melia azedarach contains Alkaloids, Carbohydrate, Phenol, Saponin, Terpenoids, Coumarin and Glycoside and absence of Amino Acid, Flavonoids, Tannins, Oxalate and Quinones.Alkaloids, Carbohydrate, Tannins, Saponins, Terpenoids and Oxalate are presentand Amino Acid, Phenol, Flavonoids, Quinones, Coumarin, Glycosides are absent in the Methanol extract.Alkaloids, Amino Acid, Phenol, Flavonoids, Terpenoids, Quinones are presentand Carbohydrates, Tannins, Saponin, Oxalate, Coumarin and Glycosides are absent in the Water extract of Melia azedarach. Anthocyanin is absent in all the solvent extracts of Melia azedarach.Acetone, Benzene, Methanol and Water extract of Melia azedarach show activity against the tested pathogens by the highest zone of inhibition Staphylococcus aureus (10 mm), Escherichia coli (9 mm), Pseudomonas aeruginosa (7 mm) and Bacillus subtilis (5 mm)for Acetone Extract of Melia azedarach, Staphylococcus aureus (6 mm), Bacillus subtilis (5 mm), Escherichia coli (5 mm) and Pseudomonas aeruginosa (4 mm)recorded. Benzene Extract of Melia azedarach resulted inStaphylococcus aureus (16 mm), Escherichia coli (13 mm), Pseudomonas aeruginosa (9 mm) and Bacillus subtilis (6 mm) and for Methanol Extract of Melia azedarach and Pseudomonas aeruginosa (6 mm) and other pathogens are resistant to the Water extract of Melia azedarach.The antioxidant activity level of Melia azedarach solvent extracts was high to Benzene followed by Acetone, Methanol and Water. The results of the study indicate that the Melia azedarach possesses phyto-constituents having antibacterial activity thus it can be utilized as a natural plant based antimicrobials.

ABSTRACT: In India, 11 species of shrimp have been found to be suitable for culture among 23 species available. Litopenaeus vannamei, Penaeus monodonand Macrobrachium rosenbergii are the major three species contributing great economy in Indian aquaculture industry.Total freshwater aquaculture production increased in 2016 compared to previous year.Aquaculture production of M. rosenbergii increased from 8,729 tonnes in 2015 to 10,152 tonnes in 2016. Giant freshwater prawn M. rosenbergii is becoming an important targeted species, as its culture is considered to be the potential source of income among farmers. This study reviews the current production status of freshwater prawn, white tail diseaseand its diagnostic methods and potential drugs available to treat the prawn diseases.

Abstract
The gene encoding VP28 protein of WSSV was cloned into pRSET B vector, and
recombinant VP28 (r-VP28) protein was produced in E. coli GJ1158 by induction with NaCl.
The r-VP28 protein was purified. The purified r-VP28 protein was injected intramuscularly in
Whiteleg shrimp, Litopenaeus vannamei, to study its tissue distribution at different time
intervals by ELISA using the antiserum raised against r-VP28, and the results showed the
presence of r-VP28 protein in different organs such as heart, hepatopancreas, gill, muscle, and
hemolymph. The r-VP28 protein was cleared from all the organs after 264 h post-injection
(hpi). Various immunological parameters such as phenoloxidase activity (PO), superoxide
anion production, activity of superoxide dismutase, and MDA content were assessed in normal
and experimentally r-VP28 protein injected shrimp at different time intervals. The mRNA
expression of ten immune-related genes was analyzed in hemocytes, hepatopancreas, and gill
by quantitative real-time PCR in order to investigate their expression to r-VP28 injection in
L. vannamei, and the results showed the upregulation of expression of immune genes such as
PO, cMnSOD, lipopolysaccharides (LPs), BGBP, hemocyanin, crustin, lectin, toll receptor,
lysozyme, and tumor necrosis factor receptor-associated factor 6 (TRAF6) in VP28-injected
shrimp. The efficacy study revealed that 86.67% survival was observed in shrimp orally
treated with r-VP28 protein after WSSV challenge without any clinical sign of WSSV
infection till the end of experimental period.

Abstract
In the present study, Response Surface Methodology (RSM) involving a central composite design
for seven factors was successfully employed to optimize the parameters for increased production of r-VP28 protein. The fermentation technique was applied to improve the yield of r-VP28 protein produced by Escherichia coli GJ1158 since Escherichia coli have been used for the production of recombinant proteins due to their growth and well known genetic characteristics. Under optimized condition r-VP28 was increased to 2.13 fold in comparison with commonly used original medium. Several parameters like temperature, pH, media concentration, time of induction and amount of inoculum were optimized to produce r-VP28 protein. Biomass concentration was measured in 41 different medium compositions for optimization of the fermentation conditions for large scale production of r-VP28. RSM was found to be effective in optimizing nutritional requirements under the optimized medium, 420.0 mg DCW/L biomass was obtained.

Abstract
An eco-friendly approach for green synthesis of Titanium nanoparticles using leaf extract of
Murraya koenigii was investigated in the present research. Fourier Transform Infrared (FTIR)
Spectroscopy and Field Emission Scanning Electron Microscopy (FESEM) analysis were used to
characterize the synthesized Titanium nanoparticles. Electron microscopy analysis of these particles
reveals the development of nanoparticles with spherical shape. The antibacterial activity of the
nanoparticles was evaluated against Escherichia coli and Pseudomonas aeruginosa. Low concentration of Titanium nanoparticles inhibited the growth both Escherichia coli and Pseudomonas aeruginosa. The particles were found to be effective against clinical pathogens such as Escherichia coli and Pseudomonas aeruginosa due to their hydroxyl bonds present and can be further used in antibiotic drugs.

Abstract
An eco-friendly approach for green synthesis of Titanium nanoparticles using leaf extract of
Citrus limon was investigated in the present research. Fourier transform infrared (FTIR) spectroscopy, field emission scanning electron microscopy and energy dispersive spectrometry (FESEM–EDS) analysis were used to characterize the synthesised titanium nanoparticles. Electron microscopy analysis of these particles reveals irregular shape of variable morphology. The antibacterial activity of the nanoparticles was evaluated against Escherichia coli and Pseudomonas aeruginosa. Low concentration of Titanium nanoparticles, inhibited the growth of Escherichia coli whereas the growth - inhibitory effects on Pseudomonas aeruginosa were mild. Minimum inhibitory concentration (MIC) value of Titanium nanoparticles against pathogenic strains of Escherichia coli and Pseudomonas aeruginosa were investigated with Ampicillin as positive control.

Abstract
Aeromonas caviae and Macrobrachium rosenbergii Nodavirus/ Extra small virus (MrNV/XSV) is
an important pathogens to giant freshwater prawn, Macrobrachium rosenbergii cultured system in
worldwide. In the present study, we examined the immunological parameters, some important immune related genes and also histopathological changes in experimentally A. caviae and MrNV/XSV injected prawn. The results obtained were compared with healthy animals. Enhanced mRNA transcripts were observed in all tested immune genes such as superoxide dismutase, prophenol oxidase, crustin, peroxinectin, anti-lipopolysaccharides and lysozyme with internal control β-actin gene in both A. caviae and MrNV/XSV injected prawn. Experimentally injected prawn showed increased prophenol oxidase activity, high levels of superoxide anion production and significant increment of clotting time than the control group. Decreased haemocyte count and superoxide dismutase were noted whereas, no significant differences in oxyhemocyanin level. Superoxide anion levels and superoxide dismutase levels showed that the activity of NADPH oxidase responsible for the release of superoxide anion increased together with a decrease in the activity of superoxide dismutase responsible for scavenging superoxide anion. After injection with A. caviae and MrNV/XSV the prawn showed mild to extensive hemocytic infiltration in the muscle often accompanied by melanization. Infiltration of haemocytes at the site of injection of A. caviae was higher when compared to infiltration at MrNV/XSV injected site.

Abstract

Abstract
The giant freshwater prawn, Macrobrachium rosenbergii, is an economically important and extensively cultured crustacean worldwide. The viral pathogens, Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) are responsible for causing severe mortalities in the hatchery and nursery phases. This study investigates the protection of postlarvae of freshwater against white tail disease (WTD) using plant extract derived from Cyanodon dactylon and the modulation of the prawn non-specific immunity. To determine the immunomodulatory effect of C. dactylon extract, the prawn was injected with plant extract and various immunological parameters were estimated. The immunological parameters such as proPO, SOD, THC and clotting time were found to be significantly higher in the plant extract-injected prawn when compared with control groups. The results of real time PCR analysis revealed up regulation on the expression proPO, SOD and lysozyme genes in MrNV and XSV challenged prawn postlarvae treated with C. dactylon extract. Infectivity experiment showed high relative per cent survival in MrNV and XSV-challenged prawn postlarvae treated with C. dactylon extract. These results strongly indicate that the administration of C. dactylon plant extract enhances immunity of the prawn. Based on the results, this study recommends that the immersion of postlarvae in C. dactylon plant extract is a potential prophylactic agent against WTD.

Abstract
Fish nodavirus (betanodavirus), a viral pathogen responsible for viral nervous necrosis (VNN) was
isolated from infected Asian sea bass (Lates calcarifer). The distribution, clearance and expression of nodavirus vaccine, on the basis of DNA vaccine (pFNCPE42 DNA-pcDNA3.1) construction, were analysed in tissues of the Asian seabass by PCR, RT-PCR, ELISA and Immunohistochemistry. Fish
immunized with a single intramuscular injection of 20 lg of the pFNCPE42-DNA vaccine showed a significant increase in the serum antibody level in the 3rd week after vaccination, compared to control eukaryotic expression vector pcDNA3.1 vaccinated fish. Results from PCR studies indicated that the vaccine-containing plasmids were distributed in heart, intestine, gill, muscle and liver 10 days after vaccination. Clearance of pFNCPE42-DNA vaccine was studied at 10, 25, 50, 75 and 100 days of post vaccination (d p.v). At 100 days p.v. pFNCPE42- DNA was cleared from muscle of vaccinated sea bass. In vitro and in vivo expression of fish nodavirus capsid protein gene (FNCP) was determined by fluorescent microscopy. Asian seabass was immunized with pFNCPE42-DNA vaccine at a dose of 20 lg per fish and were challenged with betanodavirus by intramuscular injection. The vaccinated seabass was protected from nodaviral infection and 77.33% of relative percent survival (RPS) was recorded.

Abstract
White tail disease (WTD) caused by Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) is a serious problem in prawn hatcheries. The gene for capsid protein of MrNV (MCP43) was cloned into pRSET B expression vector. The MCP43 protein was expressed as a
protein with a 6-histidine tag in Escherichia coli GJ1158 with NaCl induction. This recombinant
protein, which was used to raise the antiserum in rabbits, recognized capsid protein in different
WTD-infected post-larvae and adult prawn. Various immunological methods such as Western blot,
dot blot and ELISA techniques were employed to detect MrNV in infected samples using the antiserum raised against recombinant MCP43 of MrNV. The dot blot assay using anti-rMCP43 was found to be capable of detecting MrNV in WTD-infected post-larvae as early as at 24 h
post-infection. The antiserum raised against r-MCP43 could detect the MrNV in the infected
samples at the level of 100 pg of total protein. The capsid protein of MrNV estimated by ELISA
using anti-rMCP43 and pure r-MCP43 as a standard was found to increase gradually during the
course of infection from 24 h p.i. to moribund stage. The results of immunological diagnostic
methods employed in this study were compared with that of RT-PCR to test the efficiency of
antiserum raised against r-MCP43 for the detection of MrNV. The Western blot, dot blot and
ELISA detected all MrNV-positive coded samples as detected by RT-PCR.

Abstract
Giant freshwater prawn (Macrobrachiumrosenbergii, de Man, 1879) is an important commercial species due to its importance as food supply as well as a valuable export product. Since invertebrates lack a true adaptive immune response, stimulating M. rosenbergii for protection againstwhite tail disease (WTD) with r-MCP capsid protein is a possible approach. The effectiveness of intramuscular injection of adult freshwater prawn with r-MCP protein
and immersion of post-larvae (PL) of M. rosenbergii in r-MCP protein were investigated for their ability to protect against white tail disease. In experiment 1, the r-MCP protein injected adult prawn showed good immune response compared to PBS injected adult M. rosenbergii (mean body weight: 14.3 g). Samples were collected at different day intervals (24, 72, 120 and 168 h) to analyze the hematological and immunological parameters and to study the immune-related and HSP gene expressions with �² actin as internal control in hemocytes, gill tissue and hepatopancreas by RT-PCR and quantification of immune and HSP gene expression by quantitative
real-time PCR. In experiment II, the PL (10â��12) of M. rosenbergii were exposed to r-MCP protein by immersion for 24 h and challenged with MrNV and XSV inoculum at a volume equal to 0.1% of the total rearing medium. The samples were collected at different day intervals, i.e. 1st day, 2nd day and 5th day from treated group and 1st day, 5th day and moribund samples from MrNV and XSV infected PL group to analyze the expression of immune-related gene of M. rosenbergii by RT-PCR. Quantitative real-time PCR was employed to determine the cycle threshold (CT) value of immune gene expressionwith �² actin as host gene. The obtained percentage survival of post-larvae treated with purified r-MCP protein and challenged with MrNV and XSV was 76.03% on 15th day
p.i., 99.78% survival was recorded in normal group on 15th day p.i. and 0% survival was recorded in MrNV and XSV infected group on 10th day p.i. This suggests that the prawn immune system is able to specifically recognize and react with protein. This study shows that stimulation of the immune system of prawn with r-MCP helps to control WTD in giant freshwater prawn, M. rosenbergii.

Abstract
The indiscriminate use of pesticides and herbicides to enhance crop production has aroused great concern, because these products are likely to reach the aquatic environment, thereby posing a health concern for humans and aquatic species. Cypermethrin (CYP), a type II pyrethroid insecticide, is widely used in agriculture and for other purposes. Therefore a study was conducted for the assessment of cytotoxic, genotoxic and oxidative stress of CYP in IEG, CB, ICG, LRG and CSG cell lines at 24 h exposure. The cytotoxic effect of CYP in IEG, CB, ICG, LRG and CSG cell lines was assessed using MTT, NR, AB and CB assays. Linear correlations between each EC50 values, of CYP resulting in 50% inhibition of cytotoxicity parameters after 24 h exposure to CYP were calculated for IEG, CB, ICG, LRG and CSG cell lines using MTT, NR, AB and CB assays. Statistical analysis revealed good correlation with R2 = 0.90–0.939 for all combinations between endpoints employed. The percentage of DNA damage was assessed by comet assay in IEG, CB,
ICG, LRG and CSG cells exposed to CYP. The results of antioxidant parameters obtained show a significant increase in lipid peroxidation (LPO) level and decreased level of GSH, SOD and CAT in IEG, CB, ICG, LRG and CSG cell lines after exposure to increasing CYP in a concentration-dependent manner. This work proves that fish cell lines could be used not only for cytotoxicity and genotoxicity studies but also for studying oxidative stress when exposed to environmental contaminants such as pesticides and other pollutants.

Abstract
The present study examines the efficacy of DNA vaccine against nodavirus through oral route using CS/TPP (chitosan tripolyphosphate) nanoparticles encapsulation. The RNA2 capsid protein gene of nodavirus was used to construct DNA vaccine using pcDNA 3.1, a eukaryotic expression vector and the construct was named as pFNCPE. The size of the RNA2 capsid protein gene was 42 kDa and it was named as pFNCPE42. The CS/TPP nanoparticles were used to deliver the constructed plasmid. In vitro and in vivo expressions of FNCP (fish nodavirus capsid protein) gene were observed in sea bass kidney cell line (SISK) and in fish, respectively by fluorescent microscopy. The cytotoxicity of CS/TPP�¢ï¿½ï¿½pFNCPE42 DNA nanoparticles was evaluated by MTT assay using
fish cell line. Distribution of DNA vaccines in different tissues was studied in fish fed with the pFNCPE42-DNA encapsulated in CS/TPP nanoparticles and the expression of the gene was confirmed by PCR, RT-PCR, immunohistochemistry and ELISA analysis. The results indicate that DNA vaccine can be easily delivered into fish by feeding with CS/TPP nanoparticles. After oral vaccination Asian sea bass were challenged with nodavirus by intramuscular injection. A relative percent survival (RPS) rate of 60.0% was recorded. This study suggested that CS/TPP nanoparticles were promising carriers for plasmid DNA vaccine and might be used to vaccinate fish by oral approach.

Abstract
Fish nodavirus (betanodavirus), a viral pathogen responsible for viral nervous necrosis (VNN) was isolated from infected Asian sea bass (Lates calcarifer). The capsid protein gene of nodavirus was amplified and cloned in pRSET B, a prokaryotic expression vector. The constructwas named as r-FNCP. The size of the capsid proteinwas 42kDa and it was named as r-FNCP42. The r-FNCP42 protein was expressed as a protein with a 6-histidine tag in Escherichia coli BL21 with IPTG induction. Polyclonal antiserum was raised against this recombinant protein in rabbits and it recognized r-FNCP42 protein. The gene encoding capsid protein (FNCP-42) nucleotide sequence
was cloned and its sequence was analysed with sequences of other isolates such as Taiwan, Malaysia, Singapore and France available in the GenBank using Bioinformatics tools such as BLASTn, clustal W analysis. Homology comparison showed 98â��99% identitywith other isolates. Asian sea basswas immunized with purified r-FNCP42 mixed with complete adjuvant at a dose of 50 �¼g per fish and was challenged with nodavirus by intramuscular injection. The vaccinated sea bass was protected from nodaviral infection and 76% of relative percent survival (RPS) was recorded.

Abstract
An attempt was made to determine the replication efficiency of hepatopancreatic parvo-like virus (HPV) of shrimp in different organs of freshwater rice-field crab Paratelphusa hydrodomous (Herbst)
using bioassay, PCR, RT-PCR, ELISA, Western blot and q-PCR analyses. Another attempt was
made to use this crab as an alternative to penaeid shrimp for the large-scale production of HPV.
This crab was found to be highly susceptible to HPV by intramuscular injection. The systemic
HPV infection was confirmed by PCR and Western blot analyses in freshwater crab. The expression
of capsid protein gene in different organs of infected crab was revealed by RT-PCR analysis.
Indirect ELISA was used to quantify the capsid protein in different organs of the crab. The copy
number of HPV in different organs of the infected crab was quantified by q-PCR. The
results revealed a steady decrease in CT values in different organs of the infected crab during the
course of infection. The viral inoculum that was prepared from different organs of the infected crab caused significant mortality in post-larvae of tiger prawn, Penaeus monodon (Fabricius). The
results revealed that this rice-field crab could be used as an alternative host for HPV replication
and also for large-scale production of HPV.

Abstract
Naja naja venom was characterized by its immunochemical properties and electrophoretic pattern which revealed eight protein bands (14 kDa, 24 kDa, 29kDa, 45 kDa, 48kDa, 65 kDa, 72 kDa and 99 kDa) by SDS-PAGE in reducing condition after staining with Coomassie Brilliant Blue. The results showed that Naja venom presented high lethal activity.Whole venom antiserum or individual venom protein antiserum (14 kDa, 29 kDa, 65kDa, 72kDa and 99kDa) of venomcould recognize N.najavenombyWesternblottingandELISA, andN.naja venom presented antibody titer when assayed by ELISA. The neutralization tests showed that the polyvalent antiserum neutralized lethal activities by both in vivo and in vitro studies using mice and Vero cells. The antiserumcould neutralize the lethal activities in in-vivo and antivenomadministered after injection of cobra venom through intraperitoneal route in mice. The cocktail antiserum also could neutralize the cytotoxic activities
in Vero cell line by MTT and Neutral red assays. The results of the present study suggest that cocktail antiserumneutralizes the lethal activities in both in vitro and in vivomodels using the antiserumagainst cobra venom and its individual venom proteins serum produced in rabbits.

Abstract
Hepatopancreatic Parvovirus (HPV) causes infection in the early stages of shrimp leading to retarded growth, ultimaltely resulting in monetary loss to the shrimp farmers. To over come this situation screening of post-larvae (PL) by immunology-based diagnostics is required. Hence, the specific gene of capsid protein for HPV was cloned into pRSET B expression vector and rHCP overexpressed with 6- histidine tagged fusion protein in Escherichia coli BL21(DE3). Immunology-based methods like Western blot, dot blot and ELISA techniques were employed to detect HPV in infected samples using the antiserum raised in rabbits against recombinant HCP of HPV. The dot blot assay using anti-rHCP was found to be capable of detecting HPV in HPV infected post-larvae as early as at 24 h post infection. The antiserum could detect the HPV in the infected samples at 1 ng of total protein. HPV infection estimated by ELISA using anti-HCP and pure r-HCP as a standard was found to increase gradually during the course of infection from 24 h post infection. The sensitivity of antibody-based diagnostics employed in the present study was compared with that of PCR diagnostic method to screen the post-larvae for the detection of HPV.

Abstract
A new cell line, Channa striatus gill (CSG), derived from the gill tissue of murrel, was established and characterized. The CSG cell line was maintained in Leibovitz’s L-15 supplemented with 10% fetal bovine serum and has been subcultured more than 92 times. This cell line was able to grow in a range of temperatures from 22 to 32 C with optimal growth at 28 C. The plating efficiency was very high (52.21%) and doubling time was approximately 37 h. The gill cell line was cryopreserved at different passage levels and revived successfully with 85% survival. Polymerase chain reaction amplification of mitochondrial 16S rRNA using primer specific to C. striatus confirmed the origin of this cell line from murrel. The cell line was further characterized by immunocytochemical analysis, chromosome number, transfection and mycoplasma detection. The cytotoxicity of endosulfan was assessed in CSG cell line using apoptosis assay, comet assay, mitochondrial alteration and five other endpoints such as Rhodamine 123 uptake, 3-(4, 5- dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide, neutral red assay, Alamar Blue assay and Methylene Blue protein assay. Acute toxicity study on fish was conducted by exposing murrel for 96 h to endosulfan
under static conditions. Statistical analysis revealed good correlation with r2 = 0.972–0.997 among
the five endpoints. Linear correlations between the in vivo lethal concentration 50 (LC50) and each in vitro effective concentration 50 (EC50) were highly significant. The present study highlights the development of a new gill cell line from an air breathing fish that could be used as an alternative in vitro tools for studying pesticide toxicity in fish.

Abstract
Hepatopancreatic parvovirus (HPV) which causes infection in many species of penaeid shrimp is a serious viral pathogen in the young life stages of shrimp. An attempt was made to develop an in vitro system using C6/36 subclone of Aedes albopictus cell line for propagation of HPV. The results revealed that C6/ 36 cells were susceptible to this virus and the infected cells showed CPE in the form of vacuole formation. The results of PCR, immunocytochemistry and Western blot revealed the HPV-infection in C6/36 cell line. The RT-PCR analysis confirmed the replication of HPV in C6/36 cell line. The HPV load was quantified at different time intervals by ELISA and real time PCR, and the results showed the increase of viral load in C6/36 cell line in time course of infection. HPV propagated in C6/36 cell line was used to infect post-larvae of shrimp and the results showed that the twentieth passage of HPV propagated in C6/36 cell line caused 100% mortality in post-larvae after 6 weeks post infection (d.p.i.). The infected post-larvae showed clinical signs of reduced growth, reduced preening, muscle opacity and atrophy of hepatopancreas. The HPV-infection was confirmed by PCR. The results of the present study showed that C6/36 cell line can
be used as an in vitro model for HPV replication instead of whole animal.

Abstract
An attempt was made to determine the replication efficiency of white spot syndrome virus (WSSV)
of shrimp in different organs of freshwater ricefield crab, Paratelphusa hydrodomous (Herbst),
using bioassay, PCR, RT-PCR, ELISA, Western blot and real-time PCR analyses, and also to use
this crab instead of penaeid shrimp for the largescale production of WSSV. This crab was found
to be highly susceptible to WSSV by intramuscular injection. PCR and Western blot analyses confirmed the systemic WSSV infection in freshwater crab. The RT-PCR analysis revealed the expression of VP28 gene in different organs of infected crab. The indirect ELISA was used to quantify the VP28 protein in different organs of crab. It was found that there was a high concentration of VP28 protein in gill tissue, muscle, haemolymph and heart tissue. The copy number of WSSV in different organs of infected crab was quantified by real-time PCR, and the results revealed a steady increase in copy number in different organs of infected crab during the course of infection. The viral inoculum prepared from different organs of infected crab caused significant mortality in tiger prawn, Penaeus monodon (Fabricius). The results revealed that this crab can be used as an alternate host for WSSV replication and production.

Abstract
White tail disease (WTD) was found to be a serious problem in hatcheries and nursery ponds of freshwater prawn (Macrobrachium rosenbergii). The causative organisms have been identified as Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV). RT�PCR and immunological techniques such as Western blot and ELISA were used for early detection of MrNV and XSV in post-larval samples obtained from time-course experiments at different time intervals. Two viruses were purified from diseased post-larvae of M. rosenbergii by a combination of low and high speed centrifugation using sucrose and CsCl gradients to raise the antisera separately. One structural protein with molecular weight of 43 kDa (CP-43) was identified from
the purified preparation of MrNV, and two overlapping polypeptides of about 17 kDa (CP-17) and 16 kDa (CP- 16) were found in XSV particles by SDS-PAGE. The antisera raised against CP-43 of MrNV, CP-16 and CP17 of XSV in mice were used to detect MrNV and XSV by Western blot and ELISA. Published primers specific to MrNV and XSV were used for the early detection of these viruses by RT�PCR and nested RT�PCR. The postlarval samples collected at 3 h post infection (h p.i.) showed positive for both viruses by nested RT�PCR and negative by RT�PCR, Western blot and ELISA techniques. The samples collected at 24 h p.i. and thereafter were found to be positive for MrNV and XSV by RT�PCR, ELISA and Western blot analyses.

Synonymous codon usage bias is an inevitable phenomenon in organismic taxa, where synonymous codons occur at uneven frequencies in gene. Synonymous codon usage pattern differs between species and also among the genes of a genome. Codon usage bias is determined under various degrees by mutational bias, natural selection and other factors. In this study codon usage pattern of cytochrome c oxidase I (COI) gene of a solitary ascidian Herdmania momus was determined by analyzing the nucleotide content, codon distribution frequency, hydrophobicity, Relative Synonymous Codon Usage (RSCU), Effective Number of Codons (ENC), GC3 and Codon Adaptation Index (CAI) values. Results presented low GC content and higher ENC values indicating less bias in codon usage preference. The Grand Average of Hydropathicity (GRAVY) values and hydropathy plot confirmed
the hydrophobic nature of COI the protein.

Morphology-based taxonomical studies of ascidians in India are meagre due to lack of ascidian
taxonomist and limitations inherent in conventional system-based identification. The use of
short fragment of mitochondrial DNA sequence is proving highly useful in identifying species in
a situation where, the traditional morphology-based identification is difficult. In the present
study, two adult solitary ascidians collected from the Thoothukudi coast were morphologically
identified as Herdmania momus Savigny, 1816 and Microcosmus squamiger Michaelsen, 1927.
The genomic DNA of these ascidians was isolated, COI gene was amplified, sequenced and
submitted to the GenBank under the accession numbers KM058116, KM411616 and KJ944390.
Homology search result using BLAST showed that H. momus showed 100% matched with other
H. momus, while M. squamiger showed similarity with Pyura herdmani, a member of the same
family Pyuridae. The phylogenetic and genetic distance was maximum in interspecies than in
intraspecies. These COI sequences will allow the identification of the species through DNA
barcoding technique. Here, we report for the first time the COI gene of H. momus, Savigny 1816
from the Indian coast.
Keywords
Ascidian, Herdmania momus, Microcosmus squamiger, mitochondrial cytochrome c oxidase I (COI)

The mitochondrial cytochrome c oxidase subunit I (COI) gene, a widely accepted molecular marker for species identification and classification, has been questioned because of the presence of Numts. In this study we found the presence of Numts in the COI chromatogram of two tunicates, Polyclinum indicum and Polyclinum madrasensis belonging to the genus Polyclinum. Numts were also present in our sequence (Accession Number: KJ944391) and in other sequences belonging to genus Polyclinum in the GenBank record. The GeneBank database of genus Polyclinum contains COI-like sequences and COI pseudogenes, but no record of COI gene from Polyclinids. The prevalence of Numts in Polyclinids belonging to Tunicata, is an impediment to DNA barcoding studies of Polyclinum species.

KEYWORDS:
COI; Polyclinum; nuclear mitochondrial pseudogenes

CO1 gene sequences of 670 and 737 bps in length were obtained two colonial
ascidians, Polyclinum indicum and Didemnum candidum respectively using
universal primer. Lengths of the obtained sequences were in mitochondrial
cytochrome c oxidase subunit I gene range. Maximum GC content was observed
in the Didemnum candidum compared to Polyclinum indicum, although both
were GC rich. The translated product of these CO1 genes resulted amino
acids sequences of length 215 and 183 amino acids, with molecular weight of
23707.4 and 20073.9 Daltons for Polyclinum indicum and Didemnum candidum
respectively. Theoritical pI values and aliphatic index values supported the
instability index values, which proved the stability of the mitochondrial COI.
Two amino acids cysteine and glutamate were absent in Polyclinum indicum and
Didemnum candidum correspondingly. Further studies using amino acids of COI
gene from different ascidians may aid in developing a molecular key for the
identification of ascidians.
Keywords: Polyclinum indicum, Didemnum candidum, CO1 gene, amino acids.

The present investigation was aimed to evaluate efficacy of ethanolic extract seeds of
Nigella sativa against glucose induced cataract in goat eye lens. In the in vitro study, goat lenses were subjected to photographic evaluation and biochemical parameters such as protein, GSH, MDA, NA+/K+ ATPase and sodium and potassium were also analyzed. Photographic examination of the eyes showed that treatment with ethanolic extracts of seeds of N. sativa retarded the progression of lens opacification. Cataract lens treated with N. sativa elevated the activity of Na+ K+ ATPase, total and water soluble proteins and K+ ions to the level of normal level whereas reduced concentrations of Na+ ions. The MDA levels were significantly less in the N. sativa treated groups whereas, the level of GSH, in high glucose (55mM), compared to the normal control group was significantly low but N. sativa treated groups showed higher level of GSH. These results support the view that ethanolic extract of seeds of N. sativa as seen in this in vitro model may, counteracts the effects of glucose in inducing cataract to some extent.
Key words: Anticataract, Nigella sativa, goat lens, in vitro

Adult tunicate specimens collected from Gulf of Mannar (south east coast of India) were identified by
morphological characters as Eudistoma viride Tokioka 1955, E. microlarvum Kott, 1990 and E.ovatum
(Herdman, 1886). The mitochondrial cytochrome c oxidase subunit I (COI) gene of these species weresequenced and deposited in the GenBank (Accession Numbers: KJ944392-93, MH669162, KJ710709,MH667475 – 77, JX871396, KU360794, MH667483 – 84, KR867634, MH667485 and MH667486). Barcodes were created for the COI sequences of the 3 species (Eudistoma viride BOLD: ACQ3396, E. microlarvum BOLD: ACS6841 and E. ovatum BOLD: ACS7059) - the first record of COI gene of these species from India. Homology results using BLAST searches resulted in 100% intra specific similarities in E. microlarvum and E.ovatum species and 98-100% similarities among Eudistoma viride species. Phylogenetic analysis showed the Eudistoma species forming separate clusters. This study underlines the efficiency of molecular methods in delineating the ascidian species and this may aid extensive and systematic molecular inventory of India’s existing marine invertebrate biodiversity.

Abstract

Abstract