Clinical Performance of I-gel® and BlockBuster™ Laryngeal Mask Airway in Adult Patients during General Anesthesia: A Randomized Comparison.

Background: Supraglottic devices have revolutionized the current practice of airway management. We compared the clinical performance of a recently introduced BlockBuster™ Laryngeal mask airway with i-gel® in adult patients under general anesthesia. Methods: Following Institutional ethical clearance, the present study was conducted on 62 patients belonging to American Society of Anesthesiologists physical status 1 and 2 of either sex in the age group of 20-60 years under general anesthesia. Patients were randomly assigned to i-gel® (I) and BlockBuster™ (B) groups (31 per group). Time for successful insertion, insertion success rate, ease of insertion, oropharyngeal leak pressures (OLPs), and complications were assessed. Results: Mean insertion time of device was less in Group I (13.52 ± 2.58 s) than that of Group B (14.10 ± 2.04 s), which was neither clinically nor statistically significant (P = 0.330). OLP in Group B (24.52 ± 2.77 cm of H2O) was found to be significantly higher compared to Group I (20.81 ± 2.56 cm of H2O) with P < 0.001. Overall insertion and first attempt success was similar (i-gel® 31/31 [100%] and 29/31 [93.5%] and BlockBuster™ 31/31 [100%] and 29/31 [93.5%], respectively). Ease of insertion (P = 0.684) and complications (P = 0.782) of both the devices were comparable. Conclusions: Both the devices are useful and effective for airway management in adult under general anesthesia. Having a high OLP and comparable insertion time, risk of aspiration may be further reduced with the use of BlockBuster™ in comparison to i-gel®.

Agroecological Zone-Specific Diet Optimization for Water Buffalo (Bubalus bubalis) through Nutritional and In Vitro Fermentation Studies.

The water buffalo faces challenges in optimizing nutrition due to varying local feed resources. In response to this challenge, the current study introduces originality by addressing the lack of region-specific feeding strategies for water buffaloes. This is achieved through the formulation of 30 different diets based on locally available resources, offering a tailored approach to enhance nutritional optimization in diverse agroecological contexts. These diets were segmented into three groups of ten, each catering to the maintenance (MD1 to MD10), growth (GD1 to GD10), and lactation/production (PD1 to PD10) needs of buffaloes. Utilizing local feed ingredients, each diet was assessed for its chemical composition, in vitro gas and methane emissions, and dry matter (DM) disappearance using buffalo rumen liquor. The production diets (127 and 32.2 g/kg DM) had more protein and fats than the maintenance diets (82.0 and 21.0 g/kg DM). There was less (p < 0.05) fiber in the production diets compared to the maintenance ones. Different protein components (PB1, PB2) were lower (p < 0.05) in the maintenance diets compared to the growth and production ones, but other protein fractions (PB3, Pc) were higher (p < 0.05) in the maintenance diet. Furthermore, the growth diets had the highest amount of other protein components (PA), while the maintenance diets had the highest amount of soluble carbohydrates (586 g/kg DM), whereas the carbohydrate fraction (CB1) was highest (p < 0.05) in the production diets (187 g/kg DM), followed by the growth (129 g/kg DM) and maintenance diets (96.1 g/kg DM). On the contrary, the carbohydrate CA fraction was (p < 0.05) higher in the maintenance diets (107 g/kg DM) than in the growth (70.4 g/kg DM) and production diets (44.7 g/kg DM). The in vitro gas production over time (12, 24, and 48 h) was roughly the same for all the diets. Interestingly, certain components (ether extract, lignin, NDIN, ADIN, and PB3 and CC) of the diets seemed to reduce methane production, while others (OM, NPN, SP, PA and PB1, tCHO and CB2) increased it. In simple words, this study reveals that different diets affect gas production during digestion, signifying a significant step towards a promising future for buffalo farming through tailored, region-specific formulations.

Role of Thromboelastography Versus Coagulation Screen as a Safety Predictor in Pre-eclampsia/Eclampsia Patients Undergoing Lower-Segment Caesarean Section in Regional Anaesthesia.

PURPOSE: In this study, we aimed to correlate thromboelastography (TEG) variables versus conventional coagulation profile in all patients presenting with pre-eclampsia/eclampsia and to see whether TEG would be helpful for evaluating coagulation in parturients before regional anaesthesia. MATERIALS AND METHODS: This was a prospective study on 100 pre-eclampsia/eclampsia patients undergoing lower-segment caesarean section under regional anaesthesia. Two blood samples were collected. First sample was used for TEG measurement and second sample for laboratory tests. The following TEG data were obtained-reaction time, kinetic time, alpha angle, and maximum amplitude (MA). The following laboratory tests were obtained-haematology (haemoglobin, TLC, DLC, platelet count) and coagulation test [prothrombin time (PT), activated partial thromboplastin time (aPTT), thrombin time (TT)]. RESULT: Out of 100 patients enrolled in the study, 80 (80 %) had a normal coagulation profile, while remaining 20 (20 %) had hypocoagulation profile. The results show that TEG parameters have a good correlation with conventional coagulation profile and also showed excellent independent predictive efficacy for prediction of hypocoagulation. PT, aPTT, and TT were directly proportional to R-time and K-time and inversely proportional to alpha angle (p < 0.001). Platelet count showed a strong positive correlation with MA (p < 0.001). CONCLUSION: By giving a global picture of haemostasis, TEG can lead to improved decision-making about safety of using regional anaesthesia. Its fast feedback time makes it ideal for monitoring in a fast moving situation such as in obstetric emergency.

Low-depth shotgun sequencing resolves complete mitochondrial genome sequence of Labeo rohita.

Labeo rohita, popularly known as rohu, is a widely cultured species in whole Indian subcontinent. In the present study, we used in-silico approach to resolve complete mitochondrial genome of rohu. Low-depth shotgun sequencing using Roche 454 GS FLX (Branford, Connecticut, USA) followed by de novo assembly in CLC Genomics Workbench version 7.0.4 (Aarhus, Denmark) revealed the complete mitogenome of L. rohita to be 16 606 bp long (accession No. KR185963). It comprised of 13 protein-coding genes, 22 tRNAs, 2 rRNAs and 1 putative control region. The gene order and organization are similar to most vertebrates. The mitogenome in the present investigation has 99% similarity with that of previously reported mitogenomes of rohu and this is also evident from the phylogenetic study using maximum-likelihood (ML) tree method. This study was done to determine the feasibility, accuracy and reliability of low-depth sequence data obtained from NGS platform as compared to the Sanger sequencing. Thus, NGS technology has proven to be competent and a rapid in-silico alternative to resolve the complete mitochondrial genome sequence, thereby reducing labors and time.

Authentication of five Barilius species from Indian waters using DNA barcoding.

Authentic identification of fish species is essential for conserving them as a valuable genetic resource in our environment. DNA barcoding of living beings has become an important and ultimate tool for establishing their molecular identity. Among cyprinids, Barilius is an important genus having nearly 23 species in Indian region whose morphological identification is often difficult due to minute differences in their features. Five species collected from Indian waters and primarily identified as Opsarius bakeri (syn. Barilius bakeri), B. gatensis, B. vagra, B. bendelisis and B. ngawa were authenticated by their DNA barcoding based on mitochondrial COI gene sequences. Five individuals of each species were taken for barcode preparation by COI gene sequencing which yielded one barcode for B. ngawa, two barcodes each for O. bakeri, B. gatensis, B. bendelisis and three barcodes for B. vagra. The order of inter and intra-specific variation was estimated to know a preliminary status of variation prevailing in these cold stream fish species significant for evolution and conservation of these valued species of our ichthyofauna. Average variation within genera was found to be 13.6% with intra-specific variation ranging from 0.0% (B. ngawa) to 0.6% (B. gatensis). These distance data are in the same order found by various researchers globally using COI barcode sequences in different fish species. Phylogenetic relatedness among Barilius species and some other cyprinids validate their status of individual species as established by conventional taxonomy.

Establishment and characterization of a continuous cell line from thymus of striped snakehead, Channa striatus (Bloch 1793).

The establishment and characterization of a continuous cell line from the thymus of air-breathing fish Channa striatus are described. The cell line, designated C. striatus thymus (CST), has been subcultured over 71 times and shows optimal growth at 28°C in Leibovitz's-15 (L-15) medium supplemented with 20% fetal bovine serum. The CST cells exhibited low plating efficiency which improved with increase in seeding density. The karyotype analysis revealed that CST cells have a normal diploid karyotype with 2n = 40. Partial amplification and sequencing of two mitochondrial genes, viz. 16S ribosomal RNA (rRNA) and cytochrome oxidase I, confirmed that the cell line originated from C. striatus. CST cells were successfully transfected indicating their potential application for expression of recombinant proteins. In immunocytochemical staining, CST cells showed characteristics of epithelial cells. These cells were sensitive to extracellular products of Vibrio cholerae MTCC 3904 as well as to heavy metal mercuric chloride. The CST cell line would be a useful tool in functional genomic studies such as RNA interference and gene knockout as well as for cytotoxicity studies.

ATPase 8/6 GENE BASED GENETIC DIVERSITY ASSESSMENT OF SNAKEHEAD MURREL, Channa striata (Perciformes, Channidae).

The mitochondrial DNA (mtDNA) ATPase 8/6 gene has been used in phylogenetic as well as in phylogeographic studies along with other mtDNA markers. In this study, ATPase gene sequences were used to assess the genetic structuring and phylogeographic patterns in Channa striata. Out of 884 nucleotide positions generated in ATPase 8/6 genes, 76 were polymorphic. The study suggested 23 unique haplotypes from 67 individuals of nine populations collected from different riverine systems of India. The ATPase 8/6 sequence revealed highest haplotype as well as nucleotide diversities in Imphal River population and lowest diversities in Tapti River population. The pattern of genetic diversity and haplotype network indicated distinct mitochondrial lineages for Chaliyar population, whereas mismatch distribution strongly suggested a population expansion in mid pleistocene epoch (0.4 Mya) with distinct genetic structuring in C. striata. The baseline information on genetic variation and the population sub-structuring would facilitate conservation and management of this important snakehead murrel.

Assessment of tissue-specific effect of cadmium on antioxidant defense system and lipid peroxidation in freshwater murrel, Channa punctatus.

The effects of different concentrations of cadmium chloride on the extent of lipid peroxidation (LPO) and alterations in the antioxidant enzyme activities were studied in liver, kidney and gill tissues of freshwater murrel, Channa punctatus. The fish specimens were exposed to 6.7, 13.4 and 20.1 mg l(-1) sublethal concentrations of cadmium chloride and the oxidative stress was assessed after 24, 48, 72 and 96 h post-exposure. The biomarkers selected for the study were thiobarbituric acid reactive substances for assessing the extent of lipid peroxidation and antioxidant defense system such as reduced glutathione (GSH), glutathione reductase (GR), glutathione peroxidase (GP(X)), glutathione-S-transferase (GST), catalase (CAT) and superoxide dismutase (SOD) activities. In general, the cadmium exposure elevated the LPO in subject tissues of treated group and modulated the activities of GPx, GST, SOD, CAT, GR and level of GSH after given exposure as compared to the control. All enzymes activities, except CAT (in kidney and gills), and amount of LPO elevated significantly (P < 0.05) in treated group with respect to control in all tissues, while significant difference was not observed between the exposed concentrations and within exposure duration. The results indicated that increase in LPO level and the fluctuation in antioxidant defense system in fish could be due to cadmium-induced increase in the production of reactive oxygen species (ROS). The potential role of these parameters as biomarkers of heavy metal pollution in aquatic system is discussed.

Nucleotide variation and physical mapping of ribosomal genes using FISH in genus Tor (Pisces, Cyprinidae).

Molecular cytogenetic studies were carried out for localization of 18S and 5S ribosomal DNAs on chromosomes of three cyprinid fish species viz., T. khudree, T. mussullah and T. mosal mahanadicus using two color fluorescence in situ hybridization (FISH). All the species typically possessed 100 diploid chromosomes with minor variation in karyo-morphology. The 18S rDNA signals were observed on two pair of chromosomes in T. khudree and T. mussullah, and three pairs in T. mosal mahanadicus. The location of 18S signals also showed affinity to silver nitrate and chromomycin A3 staining. Similarly, variation in localization of 5S rDNA among the three species has been detected with the presence of FISH signals on one pair of chromosome in T. khudree and T. mussullah, and on two pairs in T. mosal mahanadicus. These molecular markers could be used as species specific markers for taxonomic identification and can further add in understanding the dynamics of genome organization and karyotypic evolution of these species. The 18S rDNA region was sequenced that generated 1811, 1810 and 1776 bp long 18S sequence in T. khudree, T. mussullah and T. mosal mahanadicus, respectively. The 18S rDNA sequence showed 95-98% identity among the subject species. Similarly, 5S sequencing generated 203 bp long fragments in these species with 100% identity in coding and 9.63% variability in non-transcribed spacer regions. The nucleotide sequence variations could be used for understanding the genetic diversity and will add new informative characters in comparative genomics. These results, in general, would enhance the value and interpretation of ecological assessment data for conservation of Tor species.

Mutagenic and genotoxic effects of carbosulfan in freshwater fish Channa punctatus (Bloch) using micronucleus assay and alkaline single-cell gel electrophoresis.

Carbosulfan insecticide is widely used in agriculture and was recently proposed for treatment against pyrethroid-resistant mosquitoes. The mutagenic and genotoxic effect of carbosulfan was carried out in fish Channa punctatus using micronucleus (MN) test and comet assay. The 96h LC(50), estimated by probit analysis in a semi-static bioassay experiment, was 0.268 mg l(-1). Based on the LC(50) value, three sub-lethal concentrations of carbosulfan (1/4th LC(50)= approximately 67 microg l(-1), 1/2nd LC(50)= approximately 134 microg l(-1) and 3/4th LC(50)= approximately 201 microg l(-1)) were selected and fishes were exposed to the said concentrations for 96h and the samplings were done at regular intervals of 24h for assessment of the MN frequencies and DNA damage. In general, significant effects (P<0.01) from both concentrations and time of exposure were observed in exposed fishes. The MN induction was highest on 96h at all the concentrations in the peripheral blood. Similar trend was observed for the DNA damage measured in terms of the percentage of tail DNA in the erythrocyte and gill cells. This study confirmed that the comet and micronucleus assays are useful tools in determining potential genotoxicity of water pollutants and might be appropriate as a part of monitoring program.

Extensive NOR site polymorphism in geographically isolated populations of Golden mahseer, Tor putitora.

Analyses of nucleolar organizer region (NOR) site polymorphism using silver staining were carried out in 72 individuals of 5 geographically isolated populations of Tor putitora from India. All Ag-NORs analyzed were constituted by rDNA that was confirmed by the presence of positive fluorescence in situ hybridization (FISH) signals. Analyses of the extra NORs among the populations confirmed the hypothesis that multi-chromosomal NOR site polymorphism is not an exception but a rule. We found a negative correlation between the mean number of extra NORs and geographical distance from the Alaknanda River population, and a significant positive correlation between the genetic distances obtained by NOR data and geographical distances among the isolated populations. These findings suggest a single geographical origin of this NOR polymorphism, from which it might have radiated to neighboring populations before their isolation. The distribution of NOR site variation was compared with the sequence variation of the ITS1 region of major rDNA (45S) and the two were found to be positively related. Different mechanisms such as transposable elements adjacent to ribosomal genes, repetitive elements which could serve as a point for chromosome exchange, amplification of minor rDNA loci, and reinsertion of extrachromosomal rDNA amplified during oogenesis have been proposed to explain the apparent NOR site polymorphism. The high dispersion of NOR site variants within and among the populations suggests that whatever transposition phenomenon is responsible for repetitive DNA or NOR jumping, it could still be active in T. putitora.

Physical mapping of rRNA gene in endangered fish Osteobrama belangeri (Valenciennes, 1844) (family: Cyprinidae).

Physical mapping of the 18S ribosomal RNA gene (rDNA) was carried out by fluorescent in situ hybridization (FISH) in the endangered freshwater fish O. belangeri. The specimens were collected from Imphal valley, Manipur, India and metaphase chromosome preparation was made using standard hypotonic treatment, methanol-acetic acid fixation and flame-drying technique followed by Giemsa, silver, CMA3 staining for complete cytogenetic characterization of the species. The diploid chromosome number was found to be 50 and the karyotype composed of 6m+16sm+12st+16t (FN=72). One active rDNA site, located on short arm of 3rd submetacentric chromosome, was mapped by FISH and confirmed by silver and CMA3 staining. The karyomorphology, chromosomal location of rDNA loci in this species and the utility of these cytogenetic markers have been discussed in the paper.

Direct and maternal (co)variance components and heritability estimates for body weights in Chokla sheep.

Estimates of (co)variance components were obtained for weights at birth, weaning and 6, 9 and 12 months of age in Chokla sheep maintained at the Central Sheep and Wool Research Institute, Avikanagar, Rajasthan, India, over a period of 21 years (1980-2000). Records of 2030 lambs descended from 150 rams and 616 ewes were used in the study. Analyses were carried out by restricted maximum likelihood (REML) fitting an animal model and ignoring or including maternal genetic or permanent environmental effects. Six different animal models were fitted for all traits. The best model was chosen after testing the improvement of the log-likelihood values. Direct heritability estimates were inflated substantially for all traits when maternal effects were ignored. Heritability estimates for weight at birth, weaning and 6, 9 and 12 months of age were 0.20, 0.18, 0.16, 0.22 and 0.23, respectively in the best models. Additive maternal and maternal permanent environmental effects were both significant at birth, accounting for 9% and 12% of phenotypic variance, respectively, but the source of maternal effects (additive versus permanent environmental) at later ages could not be clearly identified. The estimated repeatabilities across years of ewe effects on lamb body weights were 0.26, 0.14, 0.12, 0.13, and 0.15 at birth, weaning, 6, 9 and 12 months of age, respectively. These results indicate that modest rates of genetic progress are possible for all weights.

Chromosomal localization of 18S and 5S rDNA using FISH in the genus Tor (Pisces, Cyprinidae).

Dual color fluorescence in situ hybridization (FISH) was performed to study the simultaneous chromosomal localization of 18S and 5S ribosomal genes in the genus Tor for the first time. The 18S and 5S rDNAs in four Tor species were amplified, sequenced and mapped on the metaphase chromosomes. The number and distribution of 18S and 5S rDNA clusters were examined on metaphase chromosome spreads using FISH. The specimens of T. chelynoides, T. putitora and T. progeneius showed six bright fluorescent signals of 18S rDNA and T. tor exhibited ten such signals. The 5S rDNA signals were present only on one pair of chromosomes in all the four Tor species. Ag-NORs were observed on two pairs of chromosomes in T. chelynoides, T. putitora, T. progeneius and four pairs in T. tor. Comparison of the observed 18S rDNA FISH signals and Ag-NORs strongly suggested a possible inactivation of NORs localized at the telomeres of a subtelocentric and telocentric chromosome pairs in all four species. The 5S rDNA contained an identical 120 bp long coding region and 81 bp long highly divergent non-transcribed spacers in all species examined. 18S and 5S rDNA sequencing and chromosomal localization can be a useful genetic marker in species identification as well as phylogenetic and evolutionary studies.

Assessment of genotoxic and mutagenic effects of chlorpyrifos in freshwater fish Channa punctatus (Bloch) using micronucleus assay and alkaline single-cell gel electrophoresis.

Chlorpyrifos (CPF) is the single largest selling agrochemical that has been widely detected in surface waters in India. The studies on long-term genotoxic effects of CPF in different tissues of fish using genotoxic biomarkers are limited. Therefore, in the present study DNA damage by CPF in freshwater fish Channapunctatus using micronucleus (MN) and comet assays was investigated. The LC(50) - 96 h of CPF was estimated for the fish in a semi-static system. On this basis of LC(50) value sublethal and nonlethal concentrations were determined. The DNA damage was measured in lymphocytes and gill cells as the percentage of DNA in comet tails and micronuclei were scored in erythrocytes of fishes exposed to above concentrations of CPF. In general, significant effects for both the concentrations and time of exposure were observed in treated fish. It was found that MN induction in the blood was highest on day 14 at 203.0 microg/l of CPF. The highest DNA damage was observed on day 5, followed by a gradual non-linear decline in the lymphocytes and gill cells. The study indicated MN and comet assays to be sensitive and rapid methods to detect mutagenicity and genotoxicity of CPF and other pollutants in fishes.

Karyotypic diversity and evolution of seven mahseer species (Cyprinidae) from India.

Mahseer is a group of fish species that are well known as food and game fishes. The taxonomy of the mahseer species is confusing owing to the morphological variations and habitat adaptation. Detailed karyomorphological investigations have been carried out in seven species of mahseer, using karyotyping, Ag-NOR and fluorescent staining techniques. The basic diploid chromosome number (2n), in all mahseer species, was observed to be 100; however, the karyotype formula varied among the species, which were recorded as: 20m + 14sm + 22st + 44t (fundamental arm number, FN = 134) in Tor khudree; 22m + 24sm + 24st + 30t (FN = 146) in Tor mussullah; 12m + 22sm + 14st + 52t (FN = 134) in Tor putitora; 20m + 24sm + 24st + 32t (FN = 144) in Tor tor; 20m + 30sm + 24st + 26t (FN = 150) in Tor chelynoides; 20m + 20sm + 20st + 40t (FN = 140) in Tor progeneius; and 20m + 18sm + 14st + 48t (FN = 138) in Neolissochilus hexagonolepis. Silver staining of the chromosomes revealed the presence of multiple nucleolar organizer regions (NOR) in these mahseer species. The highest number of NORs was observed in T. tor (four pairs of chromosomes), whereas the other six species possessed Ag-NOR signals on only two pairs of chromosomes. Although chromomycin A(3) (CMA(3)) staining induced bright fluorescence signals on same Ag-NORs sites, with CMA(3), one additional signal was observed on the p arm of subtelocentric chromosomes in T. tor, T. chelynoides, T. progeneius and N. hexagonolepis, which may indicate the presence of inactive NOR in these species. The information on cytogenetic profile of these mahseer species is discussed in the light of cytotaxonomic implications and understanding the karyoevolution of these fish species.

Ovarian development in Labeo dyocheilus (McClelland) during active reproductive phase under captive and wild conditions.

Ovarian development in Labeo dyocheilus was assessed during active reproductive phase under ambient environmental conditions in captivity and wild. Increasing day length and water temperature seemed favourable for ovarian development in female L. dyocheilus under both conditions. Gonadosomatic index (GSI) was lower in May and higher in July in captivity (6.168 and 13.366) and wild (5.798 and 16.166) respectively Ovarian development started little bit in advance in captivity with late yolk vesicle stage oocyte in May when yolk globule stage oocytes were prominent in wild. Fully grown oocytes with germinal vesicle migration (GVM) and germinal vesicle breakdown (GVBD) stages were observed in July in both conditions. Degree of transformation of developing oogonials into advance stage oocytes was observed to be better in wild fish compared to captivity reared ones. The histophysiological changes in liver corresponded well to the progression of ovarian development. Successive granulation and vacuolization of hepatocyte cytoplasm were indicative of augmented synthetic activity and probably mobilization of energy content for oocyte growth. These observations indicated that normal ovarian development of L. dyocheilus under captivity in Tarai region of Uttarakhand would be useful for success of its seed production in captivity for stock augmentation in wild or species diversification in aquaculture.

Genotoxicity assessment of acute exposure of chlorpyrifos to freshwater fish Channa punctatus (Bloch) using micronucleus assay and alkaline single-cell gel electrophoresis.

Chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2-pyridylphosphorothioate) is one of the organophosphate pesticides widely used in agricultural practices throughout world and irreversible inhibitor of cholinesterase in all animal species. Limited efforts have been made to study acute genotoxic effects of chlorpyrifos (CPF) in different tissues of fish using genotoxic biomarkers. Therefore, the present investigation was aimed to study the induction of DNA damage by CPF in freshwater teleost fish Channapunctatus using micronucleus assay (MN assay) and alkaline single-cell gel electrophoresis (comet assay). The value of LC(50) - 96 h of CPF was determined as 811.98 microgl(-1) for C. punctatus, in a semi-static system and on the basis of LC(50) value three acute concentrations viz., 203, 406 and 609 microgl(-1) were determined. The fishes were exposed to the different concentrations of CPF for 96 h and samplings were done at regular intervals for assessment of the MN frequencies and DNA damage. In general, significant effects (P<0.01) from both concentrations and time of exposure were observed in exposed fishes. It was found that the micronucleus induction was highest on 96 h at all concentrations in the peripheral blood. Similar trend was observed for the DNA damage measured in terms of the percentage of tail DNA in the lymphocyte and gill cells. This study explored the combined use of micronucleus assay and comet assay for in vivo laboratory studies using fresh water fish for screening the genotoxic potential of xenobiotics.