An inexpensive and rapid diagnostic method for detection of SARS-CoV-2 RNA by loop-mediated isothermal amplification (LAMP).

SARS-CoV-2 is a public pandemic health concern globally. Nasopharyngeal and oropharyngeal swab samples are used for Covid-19 viral detection. Sample collection procedure was tedious and uncomfortable and unsuitable for biochemical and CBC analysis in swab samples. Biochemistry and CBC tests are key determinant in management of Covid-19 patients. We developed a LAMP test to detect viral RNA in blood samples. LAMP is required four specific primers targeting the internal transcribed S-region and loop primers for viral RNA amplification. RNA was extracted from blood samples by TRIzol method. LAMP reaction was performed at 60 °C for 1 hour and amplicons were visualized in HNB dye. No cross-reactivity was seen with HBV, HCV, and HIV infected sample. Out of 40 blood samples, 33 samples were positive for LAMP and Q-PCR analysis, one sample was positive for LAMP and negative for Q-PCR, two samples were negative for LAMP but positive for Q-PCR, and four blood samples were negative for LAMP and Q-PCR. LAMP method has an accuracy of 92.50%, with sensitivity and specificity of 94.28% and 80%, respectively. Thus, LAMP diagnostic test has proved reliable, fast, inexpensive and can be useful for detection where the limited resources available.•LAMP method is a potential tool for detection of SARS-CoV-2.•Blood samples are the key determinant for routine diagnostics as well as molecular diagnostics.•LAMP assay is an appropriate diagnostics method which offers greater simplicity, low cost, sensitivity, and specificity than other methods in molecular diagnostics.

Prevalence of Escherichia coli O157 and Salmonella spp. in solid bovine manure in California using real-time quantitative PCR.

The occurrence of Escherichia coli O157 and Salmonella spp. in solid bovine manure was investigated through a multi-county survey in California. Solid bovine manure samples (n = 91) were collected from 13 dairy farms located in multiple counties in California between June 2016 and August 2017. To quantify pathogens, DNA was extracted from bacteria in manure samples. Afterwards, the prevalence and levels of E. coli O157 and Salmonella spp. in solid bovine manure were determined by real-time quantitative PCR (qPCR). The prevalence of E. coli O157 and Salmonella spp. in solid bovine manure was 15·4 and 6·6% respectively. Escherichia coli O157 and Salmonella spp. levels in positive samples ranged from 3·1 to 5·3 log CFU per g and from positive (the population was <3 log CFU per g) to 5·2 log CFU per g respectively. Surface samples of manure piles had higher prevalence and levels of E. coli O157 and Salmonella spp. than subsurface samples, while no seasonal effects on pathogen occurrence were observed. Our results indicated that solid bovine manure is a source of E. coli O157 and Salmonella spp. and the application of untreated manure as biological soil amendments may pose potential risks to public health. SIGNIFICANCE AND IMPACT OF THE STUDY: Our findings suggested that the presence of Escherichia coli O157 and Salmonella spp. in solid bovine manure may pose potential risks if untreated manure is applied as biological soil amendments. Considering the large-scale sampling used in this study, the observations provide a holistic assessment in terms of pathogen prevalence in solid bovine manure.

Biofloc improves water, effluent quality and growth parameters of Penaeus vannamei in an intensive culture system.

Biofloc technology was evaluated with a view to analyse utilization of nitrogenous waste from the effluent and to improve water quality and growth parameters of Penaeus vannamei in intensive culture system. The experiment was carried out in two different treatment outdoor earthen ponds of 0.12 ha, one supplemented with carbon source (molasses, wheat and sugar) for biofloc formation and other was feed based control pond with a stocking density of 60 animals m-2 in duplicate for 120 days. Water, sediment and P. vannamei were sampled at regular intervals from the both set of ponds for evaluating physico-chemical parameters, nitrogen content and growth parameters, respectively. A significant reduction in the concentration of total ammonia nitrogen (TAN) and nitrite (NO2-N) were found in the biofloc pond than that of control pond. A significant low level of nitrogen was recorded in the effluents of biofloc pond in comparison to the control. In biofloc system, a significantly elevated heterotrophic bacterial count along with reduction in total Vibrio count was noticed. A significant improvement in the feed conversion efficiency (FCR) and growth parameters of P. vannamei was noticed in the biofloc pond. Growth of P. vannamei in the biofloc pond showed positive allometric pattern with an increased survival. The microbial biomass grown in biofloc consumes toxic inorganic nitrogen and converts it into useful protein, making it available for the cultured shrimp. This improved FCR and reduced the discharge of nitrogenous waste into adjacent environment, making intensive shrimp farming an eco-friendly enterprise.

Simultaneous Superior Rectus Recession and Anterior Transposition of Inferior Oblique Muscle as a Surgical Option for Traumatically Lost Inferior Rectus Muscle.

PURPOSE: To evaluate the role of simultaneous superior rectus (SR) recession and anterior transposition of inferior oblique (ATIO) muscle in patients with traumatically lost inferior rectus (IR) muscle. METHODS: Six patients with history of ocular trauma, followed by sudden onset vertical diplopia along with marked hypertropia (HT) and limitation of depression in abduction in the affected eye suggestive of IR disinsertion, were included in this prospective study. The patients were treated by simultaneous SR recession and ATIO muscle in the affected eye by limbal conjunctival approach under local anesthesia. RESULTS: Preoperatively, primary position HT of 40-50 (mean 44.16 ± 4.91) prism diopters (PD) was present in all cases which increased to 65-70 (mean 65.83 ± 5.84) PD in down and in the ipsilateral gaze along with marked limitation of depression in abduction and A pattern. On exploration, the IR could not be traced in four cases. Fibrotic muscle sheath with retracted IR was found 10-12 mm away from the limbus in rest of the two patients. ATIO (6.5 mm from the limbus) with simultaneous recession of ipsilateral SR was done under local anesthesia. At 12 weeks postoperatively, three patients were orthophoric in primary position and vertical alignment with in 4-7 PD in primary position was achieved in rest of the three patients. CONCLUSION: Simultaneous SR recession with ATIO seems to be a good alternative to achieve satisfactory vertical alignment for patients with traumatically lost inferior rectus muscle.

Congenital cranial dysinnervation disorders.

The European Neuromuscular Centre (ENMC) derived the term Congenital Cranial Dysinnervation Disorders in 2002 at an international workshop for a group of congenital neuromuscular diseases. CCDDs are congenital, non-progressive ophthalmoplegia with restriction of globe movement in one or more fields of gaze. This group of sporadic and familial strabismus syndromes was initially referred to as the 'congenital fibrosis syndromes' because it was assumed that the primary pathologic process starts in the muscles of eye motility. Over the last few decades, evidence has accumulated to support that the primary pathologic process of these disorders is neuropathic rather than myopathic. This is believed that for normal development of extra ocular muscles and for preservation of muscle fiber anatomy, normal intra-uterine development of the innervation to these muscles is essential. Congenital dysinnervation to these EOMs can lead to abnormal muscle structure depending upon the stage and the extent of such innervational defects. Over last few years new genes responsible for CCDD have been identified, permitting a better understanding of associated phenotypes, which can further lead to better classification of these disorders. Introduction of high-resolution MRI has led to detailed study of cranial nerves courses and muscles supplied by them. Thus, due to better understanding of pathophysiology and genetics of CCDDs, various treatment modalities can be developed to ensure good ocular alignment and better quality of life for patients suffering from the same.

Impact of Superior Oblique Transposition on Primary Position Deviation, a Pattern and Intorsion in Third Nerve Palsy.

PURPOSE: To evaluate the role of superior oblique transposition on primary position alignment, A pattern, and intorsion in third nerve palsy. METHODS: Ten patients with isolated, unilateral third nerve palsy were included in this prospective study. The patients were treated by conventional surgery on horizontal recti together with superior oblique transposition by Scott's procedure in the paretic eye. RESULTS: Pre-operative primary horizontal deviation was 60-80 PD (mean 70.00±7.45 PD). Pre-operative primary hypotropia was 15-22 PD (mean 18.80±2.48 PD). Mean A-pattern was 17.80±2.65 PD. All patients included in the study had some degree of objective torsion as measured by Guyton's method. One patient had grade I objective intorsion, 2 had grade II, 6 had grade III, and 1 had grade IV objective intorsion. Primary position horizontal alignment (up to ±8 PD) was achieved in 9 patients. Primary position vertical alignment (up to ±8 PD) was achieved in all 10 patients. Only 2 of 10 patients had A-pattern of 10 PD, in the remaining 8 patients it was eliminated (P<0.05). Postoperatively, 5 patients had no objective intorsion and 5 had grade I intorsion, and none of the patients had hypertropia or paradoxical eye movements. CONCLUSION: Superior oblique transposition by Scott's procedure along with conventional surgery on horizontal recti in third nerve palsy results in better horizontal and vertical alignment, and improves A-pattern and intorsion, thus leading to better binocular interaction.

Solving third-order boundary value problems with quartic splines.

In this article, we present a novel second order numerical method for solving third order boundary value problems using the quartic polynomial splines. We establish the convergence of the method. We present numerical experiments to demonstrate the efficiency of the method and validity of our second order method, which shows that present method gives better results.

Sources, distribution and risk assessment of polycyclic aromatic hydrocarbons in the mangrove sediments of Thane Creek, Maharashtra, India.

The sources, distribution and risk assessment of polycyclic aromatic hydrocarbons (PAHs) were investigated in the mangrove sediments of Trombay and Vashi, along the Thane Creek, Maharashtra, India, for a period of 6 months. The results showed that the concentration of Æ©15 PAHs ranged from 902.58 to 1643.60 and from 930.69 to 1158.30 ng g(-1) in Trombay and Vashi, respectively. Trombay showed significantly higher PAH concentration (p < 0.05) than Vashi. The four carcinogenic PAHs, (benzo(b)fluorathene, benzo(k)fluorathene, Indeno(1,2,3-cd)pyrene and dibenz(a,h)anthracene) accounted for maximum concentration of the total PAHs. Specific PAH diagnostic indices and the molecular index indicated the presence of both pyrolytic and petrogenic sources with the predominance of pyrolytic origin. A positive correlation (r = 0.736, p < 0.05) existed between the benzo(k)fluorathene level and total PAHs, suggesting the use of this compound as a potential molecular marker for PAH pollution in mangrove sediment. Assessments of potential environmental risks associated with PAHs in this study revealed that the sediment was moderately polluted with high molecular weight PAHs. The study reports the baseline data that can be used for regular monitoring of contamination level considering the heavy industrialization and urbanization along the creek and its coastal region.

Effect of different combinations of antibodies and enzyme labels on ELISA of progesterone.

In steroid immunoassays, selection of right combination of antibody and enzyme-labeled antigen determine the sensitivity and specificity of ELISA. Antibodies raised against different positions of progesterone adopting heterologous systems were reported to provide better assays for progesterone. Four different antibodies developed against progesterone-11α-hemiglutarate-BSA (P-11-HG-BSA), progesterone-11α-hemisuccinate-BSA (P-11-HS-BSA), progesterone-3-O-carboxymethyloxime-BSA (P-3-CMO-BSA), and progesterone-3-O-carboxymethyloxime-ovalbumin (P-3-CMO-ova) were tested in combination with enzyme-labeled P-11-HG, P-11-HS, progesterone-11α-carboxymethyl ether (P-11-CME), P-3-CMO, 17-hydroxyprogesterone-3-O-carboxymethyl oxime (17-P-3-CMO), and progesterone-4-carboxymethyl thioether (P-4-CMTE). These were variously labeled with penicillinase, alkaline phosphatase (ALP), and horseradish peroxidase (HRP). When antibody developed against P-11-HS-BSA was tested with P-3-CMO labeled separately with penicillinase, ALP, and HRP, the type of enzyme used had no effect on the performance of the assay. It was found that a homologous assay using P-3-CMO-ova as immunogen and P-3-CMO-HRP as label, as well as a heterologous ELISA with antibody raised against P-11-HS-BSA in combination with P-3-CMO-HRP, provided sensitive assays for progesterone. The use of 17α-hydroxy progesterone-3-O-carboxymethyl oxime-HRP with the same antibodies against P-3-CMO-BSA and P-11-HS-BSA also proved to be better than P-3-CMO-HRP. These findings implied that the sensitivity and specificity of ELISA to a great extent depended on the nature of the antibody produced, while the choice of enzyme labels could be manipulated.

Effect of orally administered azadirachtin on non-specific immune parameters of goldfish Carassius auratus (Linn. 1758) and resistance against Aeromonas hydrophila.

Modulation of the immune responses using active bio-ingredients as a possible prophylaxis measure has been novel prospect for aquaculture. The present study evaluated the effects of azadirachtin EC 25% on non-specific immune responses in goldfish Carassius auratus and resistance against pathogenic bacteria Aeromonas hydrophila. The experimental trial for effects of azadirachtin on immuno-haematoloical parameters in goldfish was conducted by feeding the various levels of azadirachtin as control T(0) (without azadirachtin), T(1) (0.1%), T(2) (0.2%), T(3) (0.4%), T(4) (0.8%) and T(5) (1.6%) for a period of 28 days. Fishes were challenged with A. hydrophila 28 days post feeding and relative percentage survival (%) was recorded over 14 days post infection. Immuno-haematoloical (total erythrocyte count, total leukocyte count, haemoglobin, packed cell volume, NBT activity, phagocytic activity, serum lysozyme activity, myeloperoxidase activity, total immunoglobulin) and serum biochemical parameters (serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT) and blood glucose) of fishes were examined at 14 and 28 days of feedings. Fish fed with azadirachtin, showed significantly (p < 0.05) enhanced TEC, TLC, Total Ig, total protein, NBT activity, serum lysozyme activity and myeloperoxidase level in different treatment groups in comparison with control group. Similarly, SGOT, SGPT and blood glucose level were found to be significantly (p < 0.05) high but PCV and Hb did not differ significantly (p > 0.05) in the treatment groups compared to control groups. Azadirachtin at the concentration of 4 g kg(-1) showed significantly (p < 0.05) higher relative percentage survival (42.60%) when compared with the control against A. hydrophila infection. This study indicated that azadirachtin EC 25% (4 g kg(-1)) showed higher NBT activity, serum lysozyme, protein profiles, leukocyte counts and resistance against A. hydrophila infection and thus, can be used as a potential immunostimulant in aquaculture.

Effect of azadirachtin on haematological and biochemical parameters of Argulus-infested goldfish Carassius auratus (Linn. 1758).

Argulosis hampers aquaculture production and alters the host physiology and growth. Azadirachtin is recognized as a potential antiparasitic agent against Argulus sp. The present study aimed to investigate the effect of different concentration of azadirachtin solution on haematological and serum biochemical parameters of Argulus-infested goldfish Carassius auratus. Ninety Argulus-infested goldfish were randomly divided into six equal groups. Fish of group 1-5 were treated with azadirachtin solution through bath of 1, 5, 10, 15 and 20 mg L(-1) as T1, T2, T3, T4 and T5, respectively, and group 6 was exposed to 2% DMSO solution without azadirachtin and considered as negative control T0(-). Along with six treatment groups, a positive control T0(+) of healthy goldfish free from Argulus infestation was also maintained. Parasitic mortality was evaluated after 3 days of consecutive bath treatment. After 7 days of post-treatment, the blood and serum were drawn from each of the treatment groups and haematological and serum biochemical parameters were evaluated. Total leucocyte count (TLC), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), blood glucose, total protein (TP), globulin, serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT) were significantly (p < 0.05) high in negative control group when compared with positive control group. It could be concluded that Argulus infestation altered marked haematological and serum biochemical parameters. However, in treated groups complete elimination of Argulus was found in T4 and T5 groups. Also significant (p < 0.05) reduction in haematological and serum biochemical parameters of all the treatment groups were recorded in comparison with negative control group. In addition, T4 and T5 groups showed significantly (p < 0.05) high superoxide dismutase (SOD), catalase, total erythrocyte count (TEC) and haemoglobin (Hb). However, higher mean corpuscular haemoglobin concentration (MCHC), blood glucose and lactate dehydrogenase (LDH) levels in T5 group revealed that higher concentration of azadirachtin have notable effects on activity of vital tissues function and physiology of the host. Argulus spp. from infested goldfish could be eliminated using bath treatment with solution of azadirachtin having concentration of 15 mg L(-1) and that also shifted haematological and serum biochemical parameters towards homeostasis.

Ocular tilt reaction, internuclear ophthalmoplegia, and torsional nystagmus following mitral commissurotomy.

Ocular tilt reaction, a type of skew deviation, and unilateral internuclear ophthalmoplegia with torsional nystagmus resulting from an ischemic event may result from unilateral disruption of otolithic pathways in the medial longitudinal fasciculus. A subset of skew deviations is known to simulate superior oblique palsy; however, none have been reported with a coexisting internuclear ophthalmoplegia. The present report documents the rare occurrence of an ocular tilt reaction simulating a left superior oblique palsy from involvement of left medial longitudinal fasciculus at the level of interstitial nucleus of Cajal.

Antiparasitic efficacy of piperine against Argulus spp. on Carassius auratus (Linn. 1758): in vitro and in vivo study.

Argulus are common aquatic ectoparasites that create one of the major threats to aquaculture due to absence of suitable therapy. Piperine, a bioactive component of Piper longum, has medicinal properties and acts as anti-inflammatory, antibacterial, and antifungal, considering eco-friendliness and cost-effectiveness. The present study aimed to evaluate antiparasitic effect of piperine against an ectoparasite Argulus spp. on Carassius auratus. Artificial Argulus infection was carried out by cohabitation method, and the fishes were selected for in vivo study when intensity of Argulus infestation was observed to be 15-20 Argulus per fish. In vitro and in vivo studies were performed at different concentration 1.0 (T (1)), 3.0 (T (2)), 5.0 (T (3)), 7.0 (T (4)), and 9.0 mg l(-1) (T (5)) of piperine solution to treat Argulus for 3 and 72 h, respectively. The acute toxicity test for piperine EC 97 % against goldfish was performed for 96 h. The 96 h median lethal concentration (LC(50)) for piperine was found to be 52.64 mg l(-1). In vitro effect of piperine solution led to 100 % mortality of Argulus at 9.0 mg l(-1) in 3 h whereas, under in vivo test, the 100 % antiparasitic efficacy of piperine solution was found at 9.0 mg l(-1) in 48 h. The EC(50) for 48 h was 9.0 mg l(-1), and thus, therapeutic index is 5.8. The results revealed that piperine at a concentration of 9.0 mg l(-1) can be used as a potential natural agent for controlling Argulus parasite.

Stoichiometric investigations of laser-ablated brass plasma.

Laser-ablated brass plasma plumes expanding in various air pressures have been studied using optical emission spectroscopy and two-dimensional imaging. The velocity of the plume front calculated from the R-t plot decreases from 1.9×10(4) m/s to ~5.5×10(3) m/s as the pressure increases from 0.01 to 10(5) Pa. The estimated higher electron temperature for Cu I (510.5 nm) transition than for Zn I (481.1 nm) may be due to differences in the heat of vaporization and vaporization temperature of copper and zinc. The electron density estimated using the Stark-broadened transition 4p(2)P(3/2)→4s(2)(2)D(5/2) of Cu I (510.5 nm) is about 10 times higher than that for transition 4s5s(3)S(1)→4s4p(3)P(2) of Zn I (481.1 nm). The appearance and enhancement of the Cu(2) (A-X) band at lower ambient pressure and formation of nanoparticle clusters have been extensively discussed. Stoichiometric and morphological study of the deposited nanoparticles on carbon tape at different ambient pressure reveals a different percentage composition of copper and of nanoparticles.

In vitro and in vivo antiparasitic activity of Azadirachtin against Argulus spp. in Carassius auratus (Linn. 1758).

Argulus is one of the most common and predominant ectoparasites which cause serious parasitic disease and is a potent carrier of viruses and bacteria in the ornamental fish industry. In recent years, organic (herbs)-based medicines are widely used to cure the disease, and neem (Sarbaroganibarini) medicine is very popular and effective throughout the world. The present study was conducted to find the effects of Azadirachtin against Argulus spp. on Carassius auratus under in vitro and in vivo conditions. The 96-h median lethal concentration (LC(50)) for Azadirachtin EC 25% against Carassius auratus was found to be 82.115 mg L(-1). The antiparasitic activity test under in vitro and in vivo was evaluated at 1 (T1), 5 (T2), 10 (T3), 15 (T4) and 20 mg L(-1) (T5) to treat Argulus for 3 h and 72 h, respectively. In vitro effect of Azadirachtin solution led to 100% mortality of Argulus at 20 and 15 mg L(-1) for 2.5 and 3 h, respectively. Whereas, under in vivo test, the 100% antiparasitic efficacy of Azadirachtin solution was found at 15 and 20 mg L(-1) for 72 and 48 h, respectively. The EC(50) for 48 h was 20 mg L(-1), and thus, therapeutic index is 4.10. The results provided evidence that Azadirachtin can be used as a potential agent for controlling Argulus.

Functional markers based molecular characterization and cloning of resistance gene analogs encoding NBS-LRR disease resistance proteins in finger millet (Eleusine coracana).

Magnaporthe grisea, the blast fungus is one of the main pathological threats to finger millet crop worldwide. A systematic search for the blast resistance gene analogs was carried out, using functional molecular markers. Three-fourths of the recognition-dependent disease resistance genes (R-genes) identified in plants encodes nucleotide binding site (NBS) leucine-rich repeat (LRR) proteins. NBS-LRR homologs have only been isolated on a limited scale from Eleusine coracana. Genomic DNA sequences sharing homology with NBS region of resistance gene analogs were isolated and characterized from resistant genotypes of finger millet using PCR based approach with primers designed from conserved regions of NBS domain. Attempts were made to identify molecular markers linked to the resistance gene and to differentiate the resistant bulk from the susceptible bulk. A total of 9 NBS-LRR and 11 EST-SSR markers generated 75.6 and 73.5% polymorphism respectively amongst 73 finger millet genotypes. NBS-5, NBS-9, NBS-3 and EST-SSR-04 markers showed a clear polymorphism which differentiated resistant genotypes from susceptible genotypes. By comparing the banding pattern of different resistant and susceptible genotypes, five DNA amplifications of NBS and EST-SSR primers (NBS-05(504,) NBS-09(711), NBS-07(688), NBS-03(509) and EST-SSR-04(241)) were identified as markers for the blast resistance in resistant genotypes. Principal coordinate plot and UPGMA analysis formed similar groups of the genotypes and placed most of the resistant genotypes together showing a high level of genetic relatedness and the susceptible genotypes were placed in different groups on the basis of differential disease score. Our results provided a clue for the cloning of finger millet blast resistance gene analogs which not only facilitate the process of plant breeding but also molecular characterization of blast resistance gene analogs from Eleusine coracana.

Characterization of nanostructured PZT prepared by chemical routes.

Nanostructured, crystalline Pb(Zr0.52Ti0.48)O3 (crystallite size 8-10 nm) ceramic powders were synthesized at low temperature by the hydroxide co-precipitation and aqueous solution method (water bath technique). The phase and surface morphology characterizations were carried out by XRD, SEM, and TEM analyses. XRD powder pattern exhibited the presence of a major tetragonal and a minor rhombohedral crystalline phase indicating the mixed phase composition in both the samples. SEM-EDS analysis revealed good homogeneity of the materials. From evolution of the Raman spectra the spectroscopic signature of the corresponding phases was determined. Detailed dielectric measurements (between 30 degrees C to 500 degrees C) were obtained to analyze a complete set of impedance-related functions such as dielectric constant (epsilon), loss factor (tan delta), dielectric modulus (M), and conductivity (sigma). Similar to the normal ferroelectric (FE) materials, the dielectric constant (epsilon) of synthesized PZTs was found to be increasing gradually with temperature and attains a maxima (epsilonmax) at the Curie temperature (Tc). The peaks in the dielectric constant plots are attributed to the phase transition associated with the change from tetragonal to rhombohedral structures. The higher dielectric constant of the PZT prepared by hydroxide co-precipitation make it more suitable for various applications.

A New Begomovirus Species Causing Leaf Curl Disease of Radish in India.

Leaf curl disease of radish (RLCD) was observed for the first time in India in commercial fields and kitchen gardens of the Varanasi District and adjoining areas of eastern Uttar Pradesh during November 2003. Infected plants exhibited typical upward and downward leaf curling, leaf distortion, reduction of leaf area, and conspicuous enations on the underside of the leaves. Incidence of RLCD was estimated to be between 10 and 40% depending on the cultivars used. Electron microscopic observation revealed typical geminate particles in infected leaf samples. The causal virus could be transmitted to radish cv. Minu Early by whiteflies (Bemisia tabaci) and grafting. Inoculated plants developed symptoms similar to those observed in naturally infected radish plants. Viral DNA was isolated from artificially inoculated symptomatic radish plants (4) followed by concentration of super-coiled DNA by alkaline denaturation (1). The presence of a geminivirus was confirmed by PCR using DNA-A degenerate primers (3), and a 1.5-kb amplified product was obtained from six artificially and three naturally infected plants. Amplification of the full-length DNA-A was achieved using a primer combination derived from sequences obtained from a 1.5-kb amplicon. Amplification of 1.3-kb DNA-ß sequences was achieved using specific primers (2) in three infected plants. Sequence analysis revealed that DNA-A (GenBank Accession No. EF 175733) contained 2,756 nt and DNA-ß contained 1,358 nt (GenBank Accession No. EF 175734). DNA-A of the causal virus shares 87.7% identity with Tomato leaf curl Bangladesh virus (GenBank Accession No. AF 188481) and 62% identity with Mungbean yellow mosaic India virus (GenBank Accession No. AF126406). The begomovirus DNA-A sequence associated with RLCD contained seven open reading frames (AV1, AV2, AC1, AC2, AC3, AC4, and AC5). The DNA-ß associated with RLCD shared the highest nucleotide sequence identity (84.9%) with DNA-ß of Tobacco leaf curl virus isolate NIB 12-1 (GenBank Accession No. AJ316033) reported from Pakistan. Despite exhaustive attempts to amplify a putative viral B-component using degenerate primers based on the intergenic region sequence of the DNA-A or sequences that are highly conserved for other begomoviruses, no DNA-B component was detected. On the basis of DNA-A sequence analysis, the ICTV species demarcation criteria of 89% sequence identity, and genome organization, the virus causing RLCD should be considered a new Begomovirus species, for which the name Radish leaf curl virus (RLCV) is proposed. To our knowledge, this is the first report of the association of a Begomovirus with a disease of radishes in India. References: (1) H. C. Birnboim and J. Doly. Nucleic Acids Res. 7:1513, 1979. (2) R. W. Briddon et al. Mol. Biotechnol. 20:315, 2002. (3) M. R. Rojas et al. Plant Dis. 77:340, 1993. (4) K. M. Srivastava et al. J. Virol. Methods 51:297, 1995.

Arsenic contamination in the Kanker district of central-east India: geology and health effects.

This paper identifies newer areas of arsenic contamination in the District Kanker, which adjoins the District Rajnandgaon where high contamination has been reported earlier. A correlation with the mobile phase episodes of arsenic contamination has been identified, which further hinges on the complex geology of the area. Arsenic concentrations in both surface and groundwater, aquatic organisms (snail and water weeds) soil and vegetation of Kanker district and its adjoining area have been reported here. The region has been found to contain an elevated level of arsenic. All segments of the ecoysystem are contaminated with arsenic at varying degrees. The levels of arsenic vary constantly depending on the season and location. An analysis of groundwater from 89 locations in the Kanker district has shown high values of arsenic, iron and manganese (mean: 144, 914 and 371 microg L(-1), respectively). The surface water of the region shows elevated levels of arsenic, which is influenced by the geological mineralised zonation. The most prevalent species in the groundwater is As(III), whereas the surface water of the rivers shows a significant contamination with the As(V) species. The analysis shows a bio-concentration of the toxic metals arsenic, nickel, copper and chromium. Higher arsenic concentrations (groundwater concentrations greater than 50 microg L(-1)) are associated with sedimentary deposits derived from volcanic rocks, hence mineral leaching appears to be the source of arsenic contamination. Higher levels of arsenic and manganese in the Kanker district have been found to cause impacts on the flora and fauna. A case study of episodic arsenical diarrhoea is presented.