Influence of selected Indian immunostimulant herbs against white spot syndrome virus (WSSV) infection in black tiger shrimp, Penaeus monodon with reference to haematological, biochemical and immunological changes

Immunostimulants are the substances, which enhance the non-specific defence mechanism and provide resistance against the invading pathogenic micro-organism. In order to increase the immunity of shrimps against the WSSV, the methanolic extracts of five different herbal medicinal plants like Cyanodon dactylon, Aegle marmelos, Tinospora cordifolia, Picrorhiza kurooa and Eclipta alba were selected and mixed thoroughly in equal proportion. The mixed extract was supplemented with various concentrations viz. 100 (A), 200 (B), 400 (C), and 800 (D) mg kg(-1) through artificial diets individually. The prepared diets (A-D) were fed individually to WSSV free healthy shrimp Penaeus monodon with an average weight of 8.0 +/- 0.5 g for 25 days. Control diet (E), devoid of herbal extract was also fed to shrimps simultaneously. After 25 days of feeding experiment, the shrimps were challenged with WSSV, which were isolated and propagated from the infected crustaceans. The shrimps succumbed to death within 7 days when fed on no herbal immunostimulant diet (E). Among the different concentrations of herbal immunostimulant supplemented diets, the shrimps fed on diet D (800 mg kg(-1)) significantly (P < 0.0001) had more survival (74%) and reduction in the viral load. Also the better performance of haematological, biochemical and immunological parameters was found in the immunostimulant incorporated diets fed shrimps. The present work revealed that the application of herbal immunostimulants will be effective against shrimp viral pathogenesis and they can be recommended for shrimp culture. (c) 2006 Published by Elsevier Ltd.

Construction and characterization of a novel recombinant single-chain variable fragment antibody against White Spot Syndrome Virus from shrimp

An antibody phage display library against White Spot Syndrome Virus (WSSV) was constructed. After four rounds of panning against WSSV, 192 out of 480 clones displayed WSSV binding activity. One of the positive clones, designated A1, had relatively higher activity specifically binding to WSSV A1-soluble, single-chain fragment variable (scFv) antibody has an affinity constant (K-aff) of 2.02 +/- 0.42 x 10(9) M-1. Dot blot assays showed that A1-soluble scFv could detect WSSV directly from shrimp hemolymph after 24-h feeding infection by WSSV. A1 scFv has potential for the development of a cheap, simple and sensitive diagnostic kit for WSSV in the field. (C) 2003 Elsevier Science B.V. All rights reserved.

Characterization of an iridovirus from the cultured pig frog Rana grylio with lethal syndrome

Three virus isolates, RGV-9506, RGV-9807 and RGV-9808, were obtained from cultured pig frogs Rana grylio undergoing lethal infections. Previously, the first isolate, RGV-9506, was shown to be an iridovirus based on ultrastructural and morphological studies. In the present study, the original isolate, along with 2 recent ones, were more extensively characterized by experimental infection studies, histopathology, electron microscopy, serological reactivity, gel electrophoresis of viral polypeptides and DNA restriction fragments, PCR amplification, and nucleic acid sequence analysis of the major capsid protein (MCP) gene. The 3 isolates were shown to be identical to each other, and very similar to FV3, the type species of the genus Ranavirus (family Iridoviridae). These results suggest that RGV should be considered a strain of FV3, and indicate that FV3-like iridoviruses are capable of causing widespread, severe disease among cultured frogs.

Purification and characterization of White Spot Syndrome Virus (WSSV) produced in an alternate host: crayfish, Cambarus clarkii

Penaeid shrimp is the natural host of White Spot Syndrome Virus (WSSV) that can cause high mortality in the infected hosts. Attempts to obtain sufficient amounts of purified intact WSSV for characterization have been unsuccessful. Using crayfish, Cambarus clarkii as a proliferation system, a large amount of infectious WSSV was reproduced and intact WSSV viral particles were purified with a new isolation medium by ultra-centrifugation. Purified WSSV particles were very sensitive to organic solvents and the detergent, Triton X-100. The size of the rod-shape, somewhat elliptical, intact WSSV was 110-130 x 260-350 mm with a long, tail-like envelope extension. The naked viral nucleocapsid was about 80 x 350 nm, and it possessed 15 spiral and cylindrical helices composed of 14 globular capsomers along its long axis, and a 'ring' structure at one terminus. Distinct WSSV genome DNA patterns were obtained when the purified genomic dsDNA of WSSV was digested with five different restriction enzymes (HindIII, XhoI, B(BamHI, SalI, and SacI). In addition, at least 13 major and distinct protein bands could be observed when purified intact WSSV viruses were separated by SDS-PAGE followed by Coomassie Brilliant R-250 staining. The estimated molecular weights of these proteins were 190, 84, 75, 69, 68, 58, 52, 44, 28, 27.5, 23, 19, and 16 kD, respectively. Both the 44 and 190 kD proteins were easily removed if the hemolymph from the: WSSV infected crayfish was transiently treated with 1%, Triton X-100 before it was subjected to gradient centrifugation, indicating that both of them are located on the surface of the viral envelope. These characteristics are consistent with WSSV isolated from the penaeid shrimp. (C) 2001 Elsevier Science B.V. All rights reserved.

Atomic configurations of dislocation core and twin boundaries in 3C-SiC studied by high-resolution electron microscopy

The defects in 3C-SiC film grown on (001) plane of Si substrate were studied using a 200 kV high-resolution electron microscope with point resolution of 0.2 nm. A posterior image processing technique, the image deconvolution, was utilized in combination with the image contrast analysis to distinguish atoms of Si from C distant from each other by 0.109 nm in the [110] projected image. The principle of the image processing technique utilized and the related image contrast theory is briefly presented. The procedures of transforming an experimental image that does not reflect the crystal structure intuitively into the structure map and of identifying Si and C atoms from the map are described. The atomic configurations for a 30 degrees partial dislocation and a microtwin have been derived at atomic level. It has been determined that the 30 degrees partial dislocation terminates in C atom and the segment of microtwin is sandwiched between two 180 degrees rotation twins. The corresponding stacking sequences are derived and atomic models are constructed according to the restored structure maps for both the 30 degrees partial dislocation and microtwin. Images were simulated based on the two models to affirm the above-mentioned results.

Microtwins and twin inclusions in the 3C-SiC epilayers grown on Si(001) by APCVD

Microtwins in the 3C-SiC films grown on Si(001) by APCVD were analyzed in detail using an X-ray four-circle diffractometer. The empty set scan shows that 3C-SiC films can grow on Si substrates epitaxially and the epitaxial relationship is revealed as (001)(3C-SiC)//(001)(Si), [111](3C-SiC)//[111](Si). Other diffractions emerged in the pole figures of the (111) 3C-SiC. We performed the (10 (1) over bar0) h-SiC and the reciprocal space mapping of the (002) plane of twins for the first time, finding that the diffractions at chi = 15.8 degrees result from not hexagonal SiC but microtwins of 3C-SiC, and twin inclusions are estimated to be 1%.

New insight into the origin of twin and grain boundary in InP

X-ray reflectivity curves show bi-crystal (twin) characteristics. Defect segregations at the twin boundary can be seen, whereas stress is relaxed at the edge of the boundary. Relaxation of the stress resulted in the formation of twins and other defects. As a result of the formation of such defects, a defect-free and stress-free zone or low defect density and small stress zone is created around the defects. Stress, chemical stoichiometry deviation and non-homogeneous distribution of impurities are the key factors that cause twins in LEC InP crystal growth. (C) 1999 Elsevier Science Ltd. All rights reserved.

Radiation-matter oscillations in attosecond polarization beats using twin color-locked noisy lights

Based on the phase-conjugate polarization interference between two one-photon processes. When the laser has broadband linewidth, the sum-frequency polarization beat (SFPB) signal shows the autocorrelation of SFPB exhibits hybrid radiation-matter detuning terahertz damping oscillation. As an attosecond ultrafast modulation process, it can be extended intrinsically to any sum-frequency of energy-levels. It hits been also found that the asymmetric behaviors of the polarization beat signals result from the unbalanced dispersion effects, (c) 2005 Elsevier B.V. All rights reserved.

Ultrafast sum-frequency polarization beats in twin Markovian stochastic correlation

An analytic closed form for the second- order or fourth- order Markovian stochastic correlation of attosecond sum- frequency polarization beat ( ASPB) can be obtained in the extremely Doppler- broadened limit. The homodyne detected ASPB signal is shown to be particularly sensitive to the statistical properties of the Markovian stochastic light. fields with arbitrary bandwidth. The physical explanation for this is that the Gaussian- amplitude. field undergoes stronger intensity. fluctuations than a chaotic. field. On the other hand, the intensity ( amplitude). fluctuations of the Gaussian- amplitude. field or the chaotic. field are always much larger than the pure phase. fluctuations of the phase-diffusion field. The field correlation has weakly influence on the ASPB signal when the laser has narrow bandwidth. In contrast, when the laser has broadband linewidth, the ASPB signal shows resonant- nonresonant cross correlation, and the sensitivities of ASPB signal to three Markovian stochastic models increase as time delay is increased. A Doppler- free precision in the measurement of the energy- level sum can be achieved with an arbitrary bandwidth. The advantage of ASPB is that the ultrafast modulation period 900as can still be improved, because the energy- level interval between ground state and excited state can be widely separated.

Twin Markovian field correlation on four-level attosecond polarization beats

Based on the phase-conjugate polarization interference between two two-photon processes, we obtained an analytic closed form for the second-order or fourth-order Markovian stochastic correlation of the four-level attosecond sum-frequency polarization beat (FASPB) in the extremely Doppler-broadened limit. The homodyne-detected FASPB signal is shown to be particularly sensitive to the statistical properties of the Markovian stochastic light fields with arbitrary bandwidth. The different roles of the amplitude fluctuations and the phase fluctuations can be understood physically in the time-domain picture. The field correlation has a weak influence on the FASPB signal when the laser has narrow bandwidth. In contrast, when the laser has broadband linewidth, the FASPB signal shows resonant-nonresonant cross-correlation, and drastic difference for three Markovian stochastic fields. The maxima of the two two-photon signals are shifted from zero time delay to the opposite direction, and the signal exhibits damping oscillation when the laser frequency is off-resonant from the two-photon transition. A Doppler-free precision in the measurement of the energy-level sum can be achieved with an arbitrary bandwidth. As an attosecond ultrafast modulation process, it can be extended intrinsically to any sum frequency of energy levels.

Coherent laser control in attosecond sum-frequency polarization beats using twin noisy driving fields

Based on the phase-conjugate polarization interference between two-pathway excitations, we obtained an analytic closed form for the second-order or fourth-order Markovian stochastic correlation of the V three-level sum-frequency polarization beat (SFPB) in attosecond scale. Novel interferometric oscillatory behavior is exposed in terms of radiation-radiation, radiation-matter, and matter-matter polarization beats. The phase-coherent control of the light beams in the SFPB is subtle. When the laser has broadband linewidth, the homodyne detected SFPB signal shows resonant-nonresonant cross correlation, a drastic difference for three Markovian stochastic fields, and the autocorrelation of the SFPB exhibits hybrid radiation-matter detuning terahertz damping oscillation. As an attosecond ultrafast modulation process, it can be extended intrinsically to any sum frequency of energy levels. It has been also found that the asymmetric behaviors of the polarization beat signals due to the unbalanced controllable dispersion effects between the two arms of interferometer do not affect the overall accuracy in case using the SFPB to measure the Doppler-free energy-level sum of two excited states.

Twin and grain boundary in InP: A synchrotron radiation study

Experimentally observed X-ray reflectivity curves show bi-crystal(twin) characteristics. The study revealed that there was defect segregation at the twin boundary. Stress was relaxed at the edge of the boundary. Relaxation of the stress resulted in formation of twin and other defects. As a result of formation of such defects, a defect-free and stress-free zone or low defect density and small stress zone is created around the defects. So a twin model was proposed to explain the experimental results. Stress(mainly thermal stress), chemical stoichiometry deviation and impurities nonhomogeneous distributions are the key factors that cause twins in LEC InP crystal growth. Twins on (111) face in LEC InP crystal were studied. Experimental evidence of above mentioned twin model and suggestions on how to get twin-free LEC InP single crystals will be discussed.

Influence of Hypericum perforatum Extract on Piglet Infected with Porcine Respiratory and Reproductive Syndrome Virus

To study the influence of Hypericum perforatum extract (HPE) on piglets infected with porcine respiratory and reproductive syndrome virus (PRRSV), enzyme-labeled immunosorbent assay (ELISA) and cytopathic effect (CPE) were used to determine in vitro whether HPE could induce swine pulmonary alveolar macrophages (PAMs) to secrete IFN-gamma, and whether PRRSV titers in PAMs were affected by the levels of HPE-induced IFN-gamma. HPE (200 mg kg(-1)) was administrated by oral gavage to piglets infected with the PRRSV in vivo to observe whether HPE affected the viremia, lung viral titers, and weight gain of piglets infected with PRRSV. The results showed that HPE was capable of inducing PAMs to produce IFN-gamma in a dose dependent manner and HPE pretreatment was capable of significantly reducing PRRSV viral titers in PAMs (P<0.01). Administration of HPE to the PRRSV-infected animals significantly (P<0.05) reduced viremia over time as compared with the PRRSV-infected animals. But there was not significant decrease in lung viral titers at day 21 post-infection between the HPE-treated animals and the PRRSV-infected control piglets. There were no significant differences in weight gain over time among the HPE-treatment animals, the normal control, and the HPE control animals. The PRRSV-infected animals caused significant (P<0.01) growth retardation as compared with the HPE controls and the normal piglets. It suggested that HPE might be an effective novel therapeutic approach to diminish the PRRSV-induced disease in swine.