Complete mitochondrial genome of critically endangered catfish Hemibagrus punctatus (Jerdon, 1849) and comparative analysis for insights into the phylogeny of hemibagrids through mitogenomic approach.

BACKGROUND: Hemibagrus punctatus (Jerdon, 1849) is a critically endangered bagrid catfish endemic to the Western Ghats of India, whose population is declining due to anthropogenic activities. The current study aims to compare the mitogenome of H. punctatus with that of other Bagrid catfishes and provide insights into their evolutionary relationships. METHODS AND RESULTS: Samples were collected from Hemmige Karnataka, India. In the present study, the mitogenome of H. punctatus was successfully assembled, and its phylogenetic relationships with other Bagridae species were studied. The total genomic DNA of samples was extracted following the phenol-chloroform isoamyl alcohol method. Samples were sequenced, and the Illumina paired-end reads were assembled to a contig length of 16,517 bp. The mitochondrial genome was annotated using MitoFish and MitoAnnotator (Iwasaki et al., 2013). A robust phylogenetic analysis employing NJ (Maximum composite likelihood) and ASAP methods supports the classification of H. punctatus within the Bagridae family, which validates the taxonomic status of this species. In conclusion, this research enriches our understanding of H. punctatus mitogenome, shedding light on its evolutionary dynamics within the Bagridae family and contributing to the broader knowledge of mitochondrial genes in the context of evolutionary biology. CONCLUSIONS: The study's findings contribute to a better understanding of the mitogenome of H. punctatus and provide insights into the evolutionary relationships within other Hemibagrids.

A rare case of giant mediastino-pleural lipoma.

Lipomas are common benign neoplastic mesenchymal tumours arising from adipose tissue anywhere in the body. Giant intra-thoracic lipoma extending across the anterior mediastinum into bilateral pleural spaces causing compression of the underlying lung resulting in respiratory embarrassment is rarely seen and reported. Giant lipomas of thorax require surgical excision as they continue to grow and cause compressive symptoms.

Description of a new Pangasius (Valenciennes, 1840) species, from the Cauvery River extends distribution range of the genus up to South Western Ghats in peninsular India.

A new species of the genus Pangasius, is described based on 17 specimens collected from the Cauvery River, India. It can be distinguished from its sister species from South and Southeast Asia, by its widely placed, small and rounded vomerine and palatine tooth plates, longer maxillary and mandibular barbels, greater vertebrae count 50 (vs. 44-48), and smaller caudal peduncle depth (6.5-8.2% SL vs. 9.89-13.09% SL). The tooth plates of the new species closely resembles that of Pangasius macronema but can be clearly distinguished from the latter by having lesser gill rakers (16-19 vs. 36-45); a smaller eye (2.4-4.4% SL vs. 5.2-9.6% SL); and larger adipose-fin base (1.5-2.9% SL vs. 0.1-1.2% SL). The mitochondrial cytochrome c oxidase (COI) gene sequence of the new species shows the genetic divergence of 3.5% and 5.1% from P. pangasius and P. silasi respectively, the two sister species found in South Asia and India. The species delimitation approaches, Poisson Tree Processes (PTP) and assemble species by automatic partitioning (ASAP) clearly resolved that the P. icaria is distinct from its sister species. Phylogenetic position of the species with its sister species was evaluated using maximum likelihood and Bayesian analysis. The discovery of this previously unknown species of genus Pangasius from the Cauvery River of peninsular India indicates important biogeographical insight that this genus migrated till the southern division of Western Ghats.

Immune gene expression in cyprinid herpesvirus-2 (CyHV-2)-sensitized peripheral blood leukocytes (PBLs) co-cultured with CyHV-2-infected goldfish fin cell line.

Goldfish is one of the preferred ornamental fish which is highly susceptible to cyprinid herpesvirus-2 (CyHV-2) infection. The present study aimed to analyse immune gene expression in a co-culture of CyHV-2-sensitized goldfish peripheral blood leukocytes (PBLs) with CyHV-2-infected fantail goldfish fin cell lines (FtGF). Goldfish were sensitized with intraperitoneal TCID50 dose (107.8±0.26/mL) of CyHV-2. After 2 weeks, PBLs were collected and co-cultured with CyHV-2-infected FtGF cells keeping both uninfected FtGF cells and PBL control groups. After 2 days of co-culture, WST-1 assay for cell proliferation was performed at 450 nm during the 2nd, 4th and 6th days of co-culture. The results showed a significant increase (p < 0.05) in cell density in CyHV-2-infected PBL and virus-infected FtGF cells during the 4th day post co-culture which confirmed effector cell generation. Expressions of few immune genes were checked taking RNA samples of CyHV-2-induced PBLs post co-culture with infected FtGF cells along with uninfected FtGF cells as control group at different time periods (2nd, 4th and 6th days) in triplicate. The results indicated increased expression of CD8α, IFNγ, b2m, MHC I, LMP 7, IL-10, IL-12 and GATA3 except Tapasin. From the above study, we concluded that goldfish showed both Th1- and Th2-mediated immune responses to CyHV-2. The current findings support the scope for further vaccine development against CyHV-2 for goldfish.

Microsatellite marker development in Spanish mackerel Scomberomorus commerson using third generation sequencing technology.

Spanish mackerel S. commerson belonging to family Scombridae, represent a group of highly commercial marine fisheries with an ever-growing demand world over. Analysing the genetic diversity of this species is of utmost importance and necessary for conservation purposes. Microsatellites are molecular tools with advantages that are ideal for population analyses. This study provides the first multiplex panel set of species-specific microsatellite loci for S. commerson that can be applied when assessing both intra- and inter population genetic variation. Microsatellite marker panels were developed in S. commerson, using Third Generation Sequencing technology in PacBio RSII, based on Single-Molecule Real-Time (SMRT). Thirty- two microsatellite loci were isolated and characterized for S. commerson, by genotyping 20 individuals each obtained from the Kochi and Veraval in the Arabian sea and Chennai along Bay of Bengal coast (n = 3). The number of alleles per locus in S. commerson varied from 4 to 17, while the mean observed and expected heterozygosities ranged from 0.656 to 0.753. The Polymorphic Information Content (PIC) were highly informative, 85% loci with PIC value 0 > 0.75. This suite of markers provides the first species specific nuclear multiplex microsatellite marker panels (32 loci) for S. commerson and thus allows assessment of different populations structures of the species across its distribution range, with more specificity. These newly developed loci have also been validated for cross transferability in another scomberid fish Scomberomorus guttatus.

Urocaridella arabianensis n. sp., a new Palaemonid shrimp (Crustacea, Decapoda, Palaemonidae) from Lakshadweep Islands, India with taxonomic comparison on the genus Urocaridella Borradaile, 1915.

A new species of the genus Urocaridella Borradaile, 1915, Urocaridella arabianensis n. sp. is described and illustrated based on fifty-five specimens collected from the reef regions of Agatti Island, Lakshadweep, Arabian Sea. Urocaridella arabianensis n. sp. is morphologically close to U. pulchella Yokes Galil, 2006, reported from the Mediterranean Sea. However, the present specimen is clearly distinguished with its rostrum length, which is 1.7 to 2.1 times as long as carapace, 10 ventral rostral teeth, carpus of 1st pereopod is 2.5 times long as palm, carpus of 2nd pereopod is 1.8 times as long as palm, presence of a spine in latero-middle of the fourth and fifth abdomens and the sixth abdominal segment is 2.3 times long as the fifth segment. Partial segment of the mitochondrial COI, 16S rRNA and nuclear gene 18S rRNA and H3 indicates that the new species is sister to a clade comprising of U. pulchella, U. antonbruunii and U. degravei. The pair wise genetic distance of COI showed great distance with the closest congeneric species.

New record of Thor hainanensis Xu amp; Li, 2014 and taxonomical remarks on Lysmata ternatensis de Man, 1902 (Decapoda: Thoridae amp; Lysmatidae) from the Lakshadweep Islands, India.

The present study reports the new record of Thor hainanensis Xu Li, 2014, on the basis of material from Agatti Islands in the Lakshadweep, India. This species was recently described from Hainan Island, China. Relationship of this species to two closely related species (T. paschalis and T. amboinensis) is discussed in this manuscript. Similarly, the present study also reports the rediscovery of other species, Lysmata ternatensis De Man, 1902 from the Lakshadweep waters with detailed taxonomic description. The present specimens were found to occur in the intertidal region in small crevices and rocks at a depth of 0.5-2.0 m. Genetic analysis using the mitochondrial COI sequences for both the species reveals close agreement for the morphological identification and showed the significant variation from the closely related species.

Aenigmachanna mahabali, a new species of troglophilic snakehead (Pisces: Channidae) from Kerala, India.

Aenigmachanna mahabali, a new species of troglophilic snakehead is described on the basis of a single specimen recovered from a well in Kerala, India, over 200km south of the type locality for the only known species in the genus. The new species can be distinguished from its congener in possessing fewer dorsal fin rays (53 vs 56-57), fewer total vertebrae (61 vs 64), fewer scales in lateral series (76 vs 83-85) and in the pectoral-fin rays being extended beyond the margin of the membrane into filaments.

Influence of the wearable posture correction sensor on head and neck posture: Sitting and standing workstations.

BACKGROUND: Flexed head and neck postures are associated with the development of neck pain in the office environment. There is little evidence regarding whether a wearable posture sensor would improve the head and neck postures of office workers. OBJECTIVE: The aim of this study was to evaluate the effect of the wearable posture sensor on the posture and physical demands on the head and neck during office work. METHODS: Nineteen participants performed a typing task with and without the wearable sensor in the sitting and standing positions. They were allowed to adjust their workstation during the experiment based on a psychophysical method. The flexion angles of the head and neck, the gravitational moment on the neck, and the positions of the workstation components were measured. RESULTS: On average, participants with the wearable sensor had 8% lower neck flexion angles and 14% lower gravitational moments on the neck than those of participants without the wearable sensor. The effect of the wearable sensor on reducing postural stress of the neck was more significant when using the standing workstation compared to the sitting workstation. CONCLUSIONS: The wearable posture sensor could be an effective tool to alleviate the postural stress of the neck in the office setting.

Establishment and characterization of a caudal fin-derived cell line, AOF, from the Oscar, Astronotus ocellatus.

Astronotus ocellatus, commonly called the oscar, is one of the popular cichlids among aquarium hobby. The present study deals with the development and characterization of a new cell line from caudal fin of A. ocellatus. The cell line was cultured in Leibovitz's L-15 medium supplemented with 10% fetal bovine serum at 28 °C. The optimum temperature and FBS concentration for cell growth were tested with temperature ranges from 20 to 37 °C and FBS concentrations of 5-20% at 28 °C. The Astronotus ocellatus fin cell line has been subcultured 45 times since its development and the modal chromosome number (2n) is 48. The cell line is composed mainly of epithelial cells as confirmed by immunocytological technique using anti-cytokeratin antibodies. The cell line was cryopreserved at different passage levels and the revival efficiency showed 80% survival rate. Partial sequence amplification and sequencing of two genes, mitochondrial 16S ribosomal RNA and cytochrome oxidase I, confirmed the origin of cell line. The cell line did not show Mycoplasma contamination. The cells showed good transfection efficiency when transfected with 2 µg of pAcGFP1-N1 expression vector. The extracellular products of fish bacterial pathogens viz., Aeromonas hydrophila and A. caviae, were cytotoxic to AOF cells but were not susceptible to Cyprinid herpes virus 2. The development of AOF cell line will have significant applications in fish virology and will prove useful to isolate pathogens in the event of sudden viral disease outbreak and for the development of vaccines and diagnostic kits.

Armrests and back support reduced biomechanical loading in the neck and upper extremities during mobile phone use.

Mobile phone use is known to be associated with musculoskeletal pain in the neck and upper extremities because of related physical risk factors, including awkward postures. A chair that provides adequate support (armrests and back support) may reduce biomechanical loading in the neck and shoulder regions. Therefore, we conducted a repeated-measures laboratory study with 20 participants (23 ±â€¯1.9 years; 10 males) to determine whether armrests and back support during mobile phone use reduced head/neck flexion, gravitational moment, and muscle activity in the neck and shoulder regions. The results showed that the chair support (armrests and back support) reduced head/neck flexion (p < 0.001), gravitational moment (p < 0.001), and muscle activity (p < 0.01) in the neck and shoulder regions significantly compared to no chair support. These results indicate that a chair with adequate support can be an effective intervention to reduce the biomechanical exposures and associated muscular pain in the neck and shoulders during mobile phone use.

Microbiological Investigation of Persistent Mortalities in Litopenaeus vannamei Grown in Low Saline Waters in India.

Abstract Microbial diseases are a serious hindrance to successful shrimp aquaculture. The Pacific white shrimp Litopenaeus vannamei is an exotic species recently introduced in India to supplement the failing aquaculture of the Asian tiger shrimp Penaeus monodon due to viral diseases. However, after a brief initial success, the aquaculture of L. vannamei is also experiencing serious problems due to microbial diseases. In this study, we conducted a microbiological investigation into the problem of persistent mortalities in selected L. vannamei farms on the southeastern coast of India. The infected shrimps were positive for the white spot syndrome virus (WSSV) by a nested PCR, though no visible white spots were present on the animals. The shrimps were heavily colonized by Vibrio parahemolyticus, which were isolated from the hepatopancreas, gills, and the body surface. The pond water, despite being low saline groundwater, harbored large numbers of V. parahemolyticus and other Vibrio species, and V. parahemolyticus isolated from L. vannamei were resistant to ß-lactam antibiotics and cephalosporins. Our results strongly suggest that the persistent mortalities of L. vannamei were due to a co-infection by V. parahemolyticus and WSSV. Received December 16, 2013; accepted February 14, 2014.

Immobilization of urease from pigeonpea (Cajanus cajan) on agar tablets and its application in urea assay.

The pigeonpea urease was immobilized on agar, a common gelling substance. The tablet strips were used as moulds to cast agar tablets of uniform shape and size. The time and temperature of solidification of agar was 6 min and 44 degrees C, respectively. The 5 % agar (w/v) and 0.019 mg protein/agar tablet yielded an optimum immobilization of 51.7%. The optimum pH was shifted through 0.2 U (from 7.3 to 7.5) towards basic side upon immobilization. The optimum temperature of soluble and immobilized urease was 30 degrees C and 60 degrees C, respectively, showing the improvement in thermal stability of urease. There was an increase in K m from 3.23 to 5.07 mM after immobilization. The half-lives of soluble and immobilized urease were 21 and 53 days, respectively, at pH 7.3 and 4 degrees C. The urea was estimated in different blood samples with the help of immobilized urease and the results were consistent with those from clinical pathology laboratory through an autoanalyzer (Zydus Co., Rome, Italy).

Boric acid and boronic acids inhibition of pigeonpea urease.

Urease from the seeds of pigeonpea was competitively inhibited by boric acid, butylboronic acid, phenylboronic acid, and 4-bromophenylboronic acid; 4-bromophenylboronic acid being the strongest inhibitor, followed by boric acid > butylboronic acid > phenylboronic acid, respectively. Urease inhibition by boric acid is maximal at acidic pH (5.0) and minimal at alkaline pH (10.0), i.e., the trigonal planar B(OH)3 form is a more effective inhibitor than the tetrahedral B(OH)4 -anionic form. Similarly, the anionic form of phenylboronic acid was least inhibiting in nature.

Fabrication of a potentiometric/amperometric bifunctional enzyme microbiosensor.

We report the fabrication and functional characterization of a needle-type bifunctional enzyme microbiosensor that has, as technical novelty, simultaneously integrated a potentiometric and amperometric detection of an enzyme-catalyzed reaction at the tip of a pulled glass micropipet. The construction involved immobilizing an enzyme onto the platinized outer tip surface using the precipitation of electrodeposition paint with direct entrapment of the biocomponent in the slowly growing polymer film. Products of enzyme-substrate reaction could then be targeted in a dual-detection mode on one hand with the covered Pt layer at the tip region as amperometric detector and on the other hand with a proton-selective liquid membrane-based potentiometric sensor inside the open pipet tip. Completing and testing bifunctional glucose microsensors demonstrated the functionality of the proposed strategy. Synchronized amperometric and potentiometric detection of the addition of a glucose standard to a buffer solution became evident by observing stepwise increases in the amperometric H2O2 oxidation current and corresponding increases in the potential of the pH-selective sensor, which translates to a local pH decrease around the tip due to hydrolysis of enzymatically formed gluconic acid.

L-Arginine increases the solubility of unfolded species of hen egg white lysozyme.

L-Arginine (L-Arg) has been widely used as an enhancer of protein renaturation. The mechanism behind its action is still not fully understood. Using hen egg white lysozyme as a model protein, we present data that clearly demonstrate the suppression of the aggregation of denatured protein by L-Arg. By chemical modification of free cysteines, a series of unfolded lysozyme species were obtained that served as models for unfolded and intermediate states during the process of oxidative refolding. An increased equilibrium solubility of unfolded species and intermediates in the presence of L-Arg seems to be its major mechanism of action.

Immobilization of pigeonpea (Cajanus cajan) urease on DEAE-cellulose paper strips for urea estimation.

Pigeonpea ( Cajanus cajan ) urease was immobilized on 1 cmx1 cm DEAE-cellulose paper strips. The optimum immobilization (51% activity) was observed at 4 degrees C, with a protein concentration of 1.0 mg/strip. The apparent optimum pH shifted from 7.3 to 6.8. Immobilized urease showed an optimal stability temperature of 67 degrees C, compared with 47 degrees C for the soluble urease. Time-dependent kinetics of the thermal inactivation of the immobilized urease were examined and found to be monophasic as compared with the soluble enzyme, which was biphasic. The Michaelis constant ( K (m)) for the DEAE-cellulose-immobilized urease was found to be 4.75 mM, 1.5 times higher than the soluble enzyme. Immobilized strips stored at 4 degrees C showed an increased half-life ( t (1/2)=150 days). There was practically no leaching of the enzyme from the immobilized strips over a period of 2 weeks. These strips were used for estimating the urea content of blood samples; the results obtained matched well with those obtained in a clinical laboratory through an Autoanalyzer(R) (Zydus Co., Rome, Italy). The easy availability of pigeonpea urease, the ease of its immobilization on DEAE-cellulose strips and the significantly lower cost of urease described in the present study makes it a suitable product for future applications in diagnostics.