An approach for dairy wastewater remediation using mixture of microalgae and biodiesel production for sustainable transportation.

The aim of this work is remediation of dairy wastewater (DWW) for biodiesel feedstock production using poly-microalgae cultures of four microalgae namely Chlorella minutissima (C. minutissima), Scenedesmus abundans (S. abundans), Nostoc muscorum (N. muscorum) and Spirulina sp. The poly-microalgae cultures were prepared as C. minutissima + N. muscorum (CN), C. minutissima + N. muscorum + Spirulina sp. (CNSS) and S. abundans + N. muscorum + Spirulina sp. (SNSS). Poly-microalgae culture CNSS cultivated on 70% DWW achieved 75.16, 61.37, 58.76, 84.48 and 84.58%, removals of biological oxygen demand (BOD), chemical oxygen demand (COD), total nitrogen (TN), total phosphorus (TP), and suspended solids (SS), respectively, at 12:12 h photoperiod that resulted into total biomass and lipid yield of 3.47 ± 0.07 g/L and 496.32± 0.065 mg/L. However, maximum biomass and lipid yields of 5.76 ± 0.06 and 1152.37 ± 0.065 mg/L were achieved by poly-microalgae culture CNSS cultivated on 70% DWW + 10 g/L of glucose at 18:6 h photoperiod. Fatty acid methyl ester (FAME) analysis shown presence of C14:0 (myristic acid) C16:0 (palmitic acid), C16:1 (palmitoleic acid), C18:0 (stearic acid), C18:2 (linoleic acid) and C18:3 (linolenic acid), it indicates that the lipids produced from poly-microalgae cultures are suitable for biodiesel production. Thus, poly-microalgae cultures could be more efficient than mono-microalgae cultures in the remediation of DWW and for biodiesel feedstock production.

Genu valgum deformity - correction by a wedgeless implantless femoral "V" osteotomy.

Coronal malalignment of the knee joint is very common in developing countries especially because of nutritional rickets. Significant valgus deformity needs to be treated surgically to improve appearance, gait and function of the patient. The purpose of this prospective study was to evaluate the results of supracondylar "V" osteotomy as a surgical technique for correction of the valgus knee deformity. This study was conducted in a tertiary level teaching hospital and 30 cases were included in the study. For all the patients deformity was assessed using ana- tomical tibiofemoral angle, mechanical axis deviation and intermalleolar distance preoperatively and post- operatively. The average age of our patients was 13.7 years and the average follow up was3.29 years (1.39-14.22 yrs). Clinically the average value of intermalleolar distance preoperatively was 16cm and 3.2 cm postperatively. Average pre-operative tibiofemoral angle was 23° and the average postoperative angle was 6 0 which was found to be statistically significant using the Paired t test (p<0.005). The average value of preoperative mechanical axis deviation was 3.1 cm which decreased to an average value of 1.1 cm postoperatively. The results with this technique have been encouraging. The advantages of this technique are low morbidity, good stability allowing early ambulation, ability to adjust alignment postoperatively by casting and no need for internal fixation. Few studies have been conducted on osteotomies that do not require internal fixation and are inherently stable. This technique has the advantage of practically no occurrence of any infection or a second surgery to remove hardware in children and adolescents. Since no specialized instrumentation, image intensifier and implants are required, it is cost effective and can be used in any primary care or district level surgical setup in a developing country like ours.

Gold-catalysed regioselective cascade cycloisomerisation reactions of aza-enediynes for the synthesis of substituted benzoisoquinoline derivatives.

Aza-enediynes underwent a facile, regioselective gold-catalysed cascade cycloisomerisation to furnish dihydrobenzo[f]isoquinoline derivatives in excellent yields. The aza-enediynes were conveniently prepared via a formal vinylic displacement reaction of allyl bromosulfones. The latter functioned as a stable and easily accessible synthetic equivalent of allenyl sulfone.

ESBL-producing Shiga-toxigenic E. coli (STEC) associated with piglet diarrhoea in India.

The present study was conducted to investigate the prevalence of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli carrying other virulence genes associated with piglet diarrhoea. Faecal samples from the piglets with history of diarrhoea were processed for isolation of E. coli and were tested for their ability of ESBL production. ESBL encoding genes and other virulence genes were detected by specific PCR, and amplicons were sequenced. Ability of transferring the ESBL genes between the enteric bacteria was tested in in vitro HGT. A total of 170 E. coli was isolated, of which 43 (25.29 %) were confirmed as ESBL producer by double disc synergy test (DDST). Altogether, 6.47 and 2.94 % isolates were positive for bla TEM and bla CTX-M-15 genes, respectively, of which 2.35 % isolates were positive for both the genes and only 0.6 % isolate was positive for the bla CTX-M gene alone. The resistance traits could not be transferred to the recipient host. Based on PCR, 2 (1.18 %) and 1 (0.59 %) isolates were recorded as Shiga-toxin-producing E. coli (STEC) and enteropathogenic E. coli (EPEC), respectively. Both the STEC (one isolate positive for stx 2 and another positive for stx 2 and hlyA) isolates belonging to serogroup O2 and NT were also positive for the bla CTX-M-15 gene. This is the first report of ESBL-producing E. coli isolate possessing the STEC gene associated with piglet diarrhoea.

Hydropericardium syndrome: current state and future developments.

Hydropericardium syndrome (HPS) is a highly infectious disease caused by fowl adenovirus serotype 4 (FAV-4) affecting poultry, especially broiler birds. The disease was initially reported from Angara Goth, Pakistan, and then from India during 1994, in the poultry belt of Jammu and Kashmir, and thereafter, from almost all parts of the country, causing heavy economic losses to the poultry industry. The disease occurs predominantly in broilers of the age group of 3-5 weeks, characterized by sudden onset of high mortality up to 80 %. The causative agent of HPS is fowl adenovirus 4, which is a member of the species Fowl Adenovirus C, genus Aviadenovirus, family Adenoviridae [60]. FAV-4 is non-enveloped and icosahedral in shape, measuring 70-90 nm in size and containing a linear dsDNA of approximately 45 kb in size as its genome. The livers of affected birds show necrotic foci and basophilic intranuclear inclusion bodies in the hepatocytes. The disease can be diagnosed from its gross and microscopic changes in the liver and by various serological tests, such as agar gel immunodiffusion, counterimmunoelectrophoresis, indirect haemagglutination, fluorescent antibody techniques, and ELISA. In the past few years, PCR has been used as a rapid diagnostic tool for the detection of fowl adenoviruses. The disease has been brought under control by the use of formalin-inactivated, attenuated or live vaccines in experimentally infected birds. Advancement in the field of computational immunology accelerates knowledge acquisition and simultaneously reduces the time and effort involved in screening potential epitopes, leading toward the development of epitope-based vaccines.

Results of titanium elastic nailing in paediatric femoral diaphyseal fractures----report of 30 cases.

OBJECTIVE: Fractures of the femur are the most incapacitating fractures. For children aged 6-14 years, there is no clear consensus as to the preferred treatment. The conventional treatment of traction and casting is no longer recommended. We report our experience in titanium elastic nailing for treatment of paediatric femoral diaphyseal fractures. METHODS: Thirty patients in the age group of 6-16 years with displaced diaphyseal femoral fractures were stabilized with titanium elastic nails. Patients were followed up clinically and radiologically for a minimum period of 1 year. The final results were evaluated using Flynn's criteria. Technical difficulties and complications associated with the procedure were also analysed. RESULTS: Overall results were excellent in 20 cases and satisfactory in 10 cases. No patient had poor result. The average hospital stay was 9 days. All the fractures healed with an average time to union of 6.86 weeks. The most common complication encountered was soft tissue irritation at the nail entry site seen in 6 cases. Clinically, lengthening was noticed in 4 cases, while no patient had shortening. Malalignment was seen in only 6 cases. Perioperative difficulties encountered were failure of closed reduction seen in 6 cases and corkscrewing of nails in one case. CONCLUSION: Titanium elastic nails are a relatively easy to use, minimally invasive, physeal-protective implant system with high rate of good and excellent outcomes in children aged 6-16 years. Technical pitfalls can be eliminated by adhering to the basic principles.

PCR-Based Detection of Extended-Spectrum ß-Lactamases (bla CTX-M-1 and bla TEM ) in Escherichia coli, Salmonella spp. and Klebsiella pneumoniae Isolated from Pigs in North Eastern India (Mizoram).

Cephalosporins are major antimicrobials used to treat serious infections. However, their effectiveness is being compromised by the emergence of extended-spectrum ß-lactamases (ESBLs). A total of 138 enteric bacteria were isolated from 53 faecal samples of pigs collected from different districts of Mizoram, of which 102 (73.91 %) were Escherichia coli, 26 (18.84 %) were Salmonella spp. and 10 (7.25 %) were Klebsiella pneumoniae. Phenotypic confirmatory test (Double Discs Synergy Test) showed that 8 (5.80 %) E. coli isolates were ESBLs producer. PCR analysis confirmed that out of the eight isolate, 7 (5.07 %) harboured bla CTX-M-1 gene and/or bla TEM gene. Of the eight positive isolates, 7 (5.07 %) and 3 (2.17 %) were found to be positive for bla CTX-M-1 gene and bla TEM gene, respectively, of which 3 (2.17 %) isolates were positive for both the genes. Only 4 (2.90 %) E. coli isolates carried bla CTX-M-1 gene alone. Agarose gel electrophoresis showed that all the isolates were carrying plasmids ranging between 0.9 and ~30 kb. Out of the seven isolates positive for bla CTX-M-1 and/or bla TEM , 2 (1.84 %) isolates were confirmed for bla CTX-M-1 gene in their plasmid. Only one E. coli isolate was found to be positive for both the genes in its plasmid. The resistance plasmid could not be transferred to a recipient by in vitro horizontal gene transfer method.

Synthesis and selective cytotoxic activity of novel hybrid chalcones against prostate cancer cells.

A new class of hybrid chalcones (17a-l &18a-l) was synthesized by Claisen-Schmidt condensation. All compounds were characterized by (1)H NMR, IR and mass spectral analysis and tested for their cytotoxic activity against PC-3 (prostate cancer), HT-29 (colon cancer), B-16 (mouse macrophages) and NCI-H460 (lung cancer) cell lines. Three compounds 18i, 18j and 18l (IC(50)=8.4, 7.9 & 5.9 µM) showed significant activity against PC-3 cell line.

Expression of rabies virus glycoprotein gene into eukaryotic system and determination of potential T-cell epitopes.

The present study was undertaken to clone, express rabies virus glycoprotein (RVG) and to identify potential T-cell epitopes on it. RVG gene (1590 bp) was amplified using gene specific primers. The amplified product was cloned into pTZ57R/T cloning vector by TA cloning. RVG gene was subcloned into pcDNA3.1 (+) expression vector. In this study, cloning and expression of rabies virus glycoprotein gene was done under CMV promoter and an expression construct (pcDNA.RVG) was prepared and clones were confirmed by restriction digestion, colony PCR and nucleotide sequencing. The expression construct was further characterized by western blotting and indirect fluorescent antibody test (IFAT). In silico analysis of this protein was done to find out potential antigenic sites so that it can be further evaluated for its potential as candidate for epitope vaccine against rabies.

Effects of various abiotic factors on biomass growth and lipid yield of Chlorella minutissima for sustainable biodiesel production.

In this study, effects of different abiotic factors were studied on biomass and lipid yield of green microalga Chlorella minutissima (C. minutissima) Various concentrations of abiotic factors like nitrogen, phosphorus, glucose, iron, zinc, different values of pH, temperature, light intensity and different photoperiods were observed on the biomass growth and lipid yield of C. minutissima cultivated with modified CHU-13 medium. Initially, three cultivation media namely, Bold's basal medium (BBM), modified CHU-13 and blue-green-11 (BG-11) were used to culture C. minutissima in batch mode. Microalga cultivated with modified CHU-13 medium resulted in maximum biomass and lipid yield of 970 ± 0.21 and 356.63 ± 0.51 mg/L, respectively. To maximize biomass and lipid yield of microalga further, it was cultivated with modified CHU-13 medium and variation of above mentioned abiotic factors was done. Different biomass and lipid yields were achieved for different abiotic factors varied. Highest biomass of 1840.49 ± 0.62 mg/L was achieved with 12 g of glucose containing medium and highest lipid yield of 579.86 ± 0.76 mg/L was achieved with 0.3 g of nitrogen containing medium. GC-MS analysis of biodiesel obtained from C. minutissima biomass cultivated with modified CHU-13 medium shown the presence of C14:0, C16:0, C16:1, C18:0, C18:1, C18:2, C18:3, C20:0, C20:1 and C22:0. Properties of biodiesel obtained from C. minutissima were found in compliance with ASTM-6751-02 and European biodiesel standards EN14214. These results suggest that C. minutissima can be used as a potential biodiesel feedstock for microalgal biodiesel production.

A novel gallium bisaminothiolate complex as a myocardial perfusion imaging agent.

The development of new myocardial perfusion imaging agents for positron emission tomography (PET) may improve the resolution and quantitation of changes in regional myocardial perfusion measurement. It is known that a (68)Ge/(68)Ga generator can provide a convenient source of PET tracers because of the long physical half-life of (68)Ge (271 days). A new ligand, 7,8-dithia-16,24-diaza-trispiro[5.2.5.2.5.3]pentacosa-15,24-diene, which consists of a N(2)S(2)-chelating core incorporated into three cyclohexyl rings, was prepared. To test feasibility and potential utility, the N(2)S(2) ligand was successfully labeled and tested with (67)Ga (half-life=3.26 day; gamma=93.3, 184.6 and 300.2 keV), which showed >92% radiochemical purity. The corresponding "cold" Ga complex was synthesized, and its structure containing a pyramidal N(2)S(2) chloride core was elucidated with X-ray crystallography. In vivo biodistribution of this novel (67)Ga complex, evaluated in normal rats, exhibited excellent heart uptake and retention, with 2.1% and 0.9% initial dose/organ at 2 and 60 min, respectively, after an intravenous injection. Autoradiography was performed in normal rats and in rats that had the left anterior descending coronary artery permanently ligated surgically. Autoradiography showed an even uptake of activity in the normal heart, and there was a distinctively lower uptake in the damaged side of the surgically modified heart. In conclusion, the new N(2)S(2) ligand was readily prepared and labeled with radioactive (67)Ga. Biodistribution in rats revealed high initial heart uptake and relatively high retention reflecting regional myocardial perfusion.

Asymmetric Reaction of p-Quinone Diimide: Organocatalyzed Michael Addition of α-Cyanoacetates.

Hitherto unknown catalytic enantioselective transformation of p-quinone diimides is achieved using chiral bifunctional organic molecules. Bifunctional thiourea compounds catalyze the Michael addition of cyanoacetates with excellent yields and enantioselectivities. The initially formed Michael adducts undergo cyclization to yield functionally rich, fused cyclic imidines bearing a quaternary benzylic chiral center. Density functional theory calculations of the competing transition states (TSs) were carried out to explain the observed stereochemical outcome.

New diphenylacetylenes as probes for positron emission tomographic imaging of amyloid plaques.

A series of 18F fluoropegylated diphenylacetylenes as probes for binding to Abeta plaques were successfully prepared. These relatively rigid acetylenes, 12a, 12b, 14a, and 14b, displayed high binding affinities in postmortem AD brain homogenates (Ki ranging from 1.2 to 2.9 nM). In vivo biodistribution in normal mice exhibited excellent initial brain penetrations (4.42, 4.55, 5.41, and 6.78% dose/g at 2 min for [18F]12a, 12b, 14a, and 14b, respectively). [18F]12b and [18F]14b, with a longer fluoropegylated unit, that is, n=3, showed faster brain washout at 30 min postinjection (0.42 and 1.57% dose/g) as compared to the shorter fluoropegylated chain ligands, that is, [18F]12a and [18F]14a (1.03 and 3.69% dose/g). Autoradiography and homogenate binding confirmed the high binding signal due to Abeta plaques. These preliminary results suggest that the novel diphenylacetylenes may be potentially useful for imaging of Abeta plaques in the brain of patients with Alzheimer's disease.

Phycoremediation potential of marine microalga Tetraselmis indica on secondary treated domestic sewage for nutrient removal and biodiesel production.

Marine microalga Tetraselmis indica (T. indica) was cultivated in secondary treated domestic sewage (STDS) in batch mode. Optimization studies showed that after 14 days of cultivation period, highest biomass yield reached was 0.88 ± 0.04 g/L at the optimum temperature of 27 ± 1 °C and light intensity of 135 µmol m-2 s-1. T. indica removed about 60.93% phosphate, 78.46% nitrate, 72.94% chemical oxygen demand (COD), 73.17% biological oxygen demand (BOD), 98.90% total dissolved solids (TDS) and heavy metals (83.11% Cd, 55.67% Ca, 45.12% Cu, 13.67% Mn, 50.88% Pb, and 98.92% Al) from STDS. The level of electrical conductivity was reduced to 0.0974 ± 0.045 dS/m. The fatty acid methyl ester (FAME) profile showed the presence of palmitic acid (12.91%), oleic acid (35.94%), linoleic acid (14.89%) and eicosanoic acid (12.34%). This study indicates the potential of T. indica for removal of pollutants from STDS and also its capability of biodiesel production.

Studies on diagnostic potential of secretory antigens of Mycobacterium bovis.

Culture filtrate proteins (CFPs) of M. bovis were produced by culturing the organism for a period of 3 weeks. These CFPs elicited a good delayed hypersensitivity (DTH) reaction in cattle and guinea pig. However, CFPs showed least cross-reactivity when tested in guinea pigs sensitized with M. bovis BCG and M. phlei. CFPs were found to be well recognized by T-cells of single intradermal test (SIDT) positive cattle and were also sero-reactive. Lymphocyte transformation assay revealed the culture filtrate antigens as major T-cell antigens. CFPs induced a good interferon gamma release as compared to conventional antigen, PPD, thereby highlighting their diagnostic capabilities.

Pathology and molecular diagnosis of classical swine fever in Mizoram.

AIM: Clinical histopathological and molecular diagnosis of classical swine fever disease in pigs of Mizoram. MATERIALS AND METHODS: Totally, 31 clinically suspected pigs from 6 districts of Mizoram were examined, and clinical symptoms were recorded. Detailed post mortem examination of all the 31 dead animals was conducted, and gross changes were recorded. Tissue samples were collected for histopathological examination and molecular diagnosis. The collected tissues (tonsil, lymph nodes, spleen) were also processed for RNA extraction. Reverse transcription polymerase chain reaction (RT-PCR) was performed to detect the specific gene fragments of classical swine fever virus (CSFV). RESULTS: Clinical examination of all the 31 suspected pigs revealed typical clinical signs of CSF. All the animals also showed typical gross and microscopic lesions of CSF. RT-PCR on tissue samples amplified the 421bp, 449bp and 735bp region of 5´NCR, non-structural protein 5B and E(rns) gene regions of CSFV, respectively. Nested PCR for internal region of E2 gene also amplified the expected product of 271bp using PCR product of whole E2 region as template DNA. CONCLUSION: CSF is highly endemic disease in Mizoram. The viral strains circulating in this region are highly virulent. The disease can be diagnosed specifically using RT-PCR.

Detection and characterization of extended-spectrum ß-lactamases (blaCTX-M-1 and blaSHV ) producing Escherichia coli, Salmonella spp. and Klebsiella pneumoniae isolated from humans in Mizoram.

AIM: The present study was conducted to isolate and characterize the extended spectrum ß-lactamases (ESBLs) producing enteric bacteria in human beings in Mizoram, India. MATERIALS AND METHODS: Fecal samples were collected from human beings with or without the history of diarrhea from different hospitals of Mizoram. Samples were processed for isolation and identification of Escherichia coli, Salmonella and Klebsiella pneumoniae. All the isolates were subjected to antibiotic sensitivity assays. Phenotypically, ESBLs production ability was determined by double discs synergy test (DDST) method. ESBLs producing isolates were subjected to polymerase chain reaction (PCR) for detection of ESBLs genes. Plasmids were cured by acridine orange. Transfer of resistance from a donor to recipient strains was done by in vitro horizontal method. RESULTS: A total of 414 enteric bacteria were isolated from 180 fecal samples (113 were from diarrheic patients and 67 were from non-diarrheic patients), of which 333 (80.44%), 52 (12.56%), and 29 (7.00%) were E. coli, K. pneumoniae and Salmonella spp., respectively. Double discs synergy test (DDST) exhibited 72 (21.62%) E. coli, 12 (23.08%) K. pneumoniae and 4 (13.79%) Salmonella spp. were ESBLs producers. Altogether, 24 (13.04%) isolates were found to be positive for at least one resistance genes under this study. A total of 36 (8.70%) E. coli, 4 (0.97%) K. pneumoniae and 2 (0.48%) Salmonella spp. were found to be positive for blaCTX-M-1 gene by PCR. Similarly, 5 (1.21%) E. coli and 4 (0.97%) K. pneumoniae isolates were found to be positive for blaSHV gene. A total of 3 (0.72%) K. pneumoniae isolates were recorded as positive for both blaCTX-M-1 and blaSHV genes. All the isolates were carrying plasmids ranging between 0.9 kb and ~30 kb. The resistance plasmid could not be transferred to a recipient by in vitro horizontal gene transfer method. CONCLUSION: ESBLs producing enteric bacteria are circulating in human population in North Eastern Region of India. Indiscriminate use of antibiotics should be avoided to control the menace of multidrug resistance bacteria in the environment, animals, and human beings.

Studies on structural and immunogenic polypeptides of hydropericardium syndrome virus by SDS-PAGE and western blotting.

The polypeptide pattern of a local isolate of a virus causing hydropericardium syndrome was analyzed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. A total of 12 polypeptides ranging in molecular weight between 13.8 and 110.0 kDa were observed. Western blot analysis of structural polypeptides revealed seven immunogenic polypeptides ranging in molecular weight between 15.8 and 110.0 kDa.

Isolation of etiological agent of hydropericardium syndrome in chicken embryo liver cell culture and its serological characterization.

The virus causing hydropericardium syndrome was isolated in chicken embryo liver (CEL) cell culture from livers obtained from naturally infected broilers. The cytopathic effects characterized by rounding and degeneration of cells were visible 36 hr post infection in first passage. At 4th passage level, the infectivity titre was 5.24 log10 TCID50/ml. In May-Grunwald and Giemsa stained cells, basophilic intranuclear inclusions ('bird eye' inclusion), typical of aviadenovirus infection, were observed. The specificity of inclusion was confirmed by indirect immunofluorescence. Various serological tests, such as agar gel precipitation test, counter immuno electrophoresis, micro serum neutralization test and enzyme linked immunosorbent assay were also standardized to confirm the isolation of etiological agent of hydropericardium syndrome in CEL cell culture and to diagnose the disease in poultry.