Effect of an immunomodulator on the efficacy of an attenuated vaccine against avian pneumovirus in turkeys

Since 1997, avian pneumovirus (APV) has caused estimated annual losses of $15 million to the Minnesota turkey industry. In order to develop an attenuated live vaccine against APV, we serially passaged a Minnesota isolate of APV (APV/MN/turkey/1-a/97) in vitro in cell cultures for 41 passages. Laboratory experiments with this high-passage virus (P41) indicated that the attenuated virus provided immunogenic protection to turkeys against challenge with virulent APV, although some birds showed mild to moderate dinical signs after inoculation. To reduce the residual pathogenicity of P41, while maintaining its immunogenicity, we decided to vaccinate turkeys with P41 in the presence of an immunomodulator, S-28828 (1-n-butyl-2-ethoxymethyl-1H-imidazo[4,5-c]quinolin-4-amine-hydrochloride), which is a potent cytokine inducer. The combined inoculation of S-28828 (5 mg/kg body weight) and P41 resulted in a significant reduction in the incidence of virus-induced clinical signs in comparison with birds that received P41 without immunomodulator (P < 0.05). Only 17% of birds inoculated with S-28828 + APV P41 showed mild respiratory symptoms at 5 days postinoculation as compared with 46% of the vaccinated turkeys that did not receive S-28828. Vaccination with either P41 or with P41 + S-28828 protected turkeys against dinical signs and viral replication after challenge with virulent APV. These results indicate that immunomodulators, such as S-28828, may act as good vaccine adjuvants that can reduce the pathogenicity but maintain the immunogenicity of partially attenuated vaccines.     

Serological Methods for Detection of Polymyxa graminis, an Obligate Root Parasite and Vector of Plant Viruses

A purification procedure was developed to separate Polymyxa graminisresting spores from sorghum root materials. The spores were used as im-munogen to produce a polyclonal antiserum. In a direct antigen coating enzyme-linked immunosorbent assay (DAC ELISA), the antiserum could detect one sporosorus per well of the ELISA plate. In spiked root samples, the procedure detected one sporosorus per mg of dried sorghum roots. The majority of isolates of P. graminis from Europe, North America, and India reacted strongly with the antiserum. Interestingly, P. graminis isolates from the state of Rajasthan (northern India), from Pakistan, and an isolate from Senegal (West Africa) reacted weakly with the antiserum. The cross-reactivity of the serum with P. betae isolates from Belgium and Turkey was about 40% of that observed for the homologous isolate. There was no reaction with common fungi infecting roots or with the obligate parasite Olpidium brassicae. However, two isolates of Spongospora sub-terranea gave an absorbance similar to that observed with the homologous antigen. The DAC ELISA procedure was successfully used to detect various stages in the life cycle of P. graminis and to detect infection that occurred under natural and controlled environments. A simple procedure to conjugate antibodies to fluorescein 5-isothiocyanate (FITC) is described. Resting spores could be detected in root sections by using FITC-labeled antibodies. The potential for application of the two serological techniques for studying the epidemiology of peanut clump disease and for the characterization of Polymyxa isolates from various geographical origins is discussed.     

Insecticidal action of the polyphenolic rich fractions from the stem bark of Streblus asper on Dysdercus cingulatus

Extracts from the stem bark of Streblus asper possess insecticidal activity against the fifth instar of Dysdercus cingulatus. Methanolic extract showed an LC(50) value of 5.56 microg/insect. Partition with chloroform increased the insecticidal activity (LC(50) 2.01 microg/insect). Three polyphenolic rich fractions were obtained from silica-gel column chromatography of the chloroform fraction and found to have noteworthy insecticidal activity (LC(50): 1.82 microg, 2.70 microg, 2.26 microg/insect) by topical application. This may provide a useful beginning for the development of biopesticides.     

The stress of weaning influences serum levels of acute-phase proteins, iron-binding proteins, inflammatory cytokines, cortisol, and leukocyte subsets in Holstein calves

The purpose of our study was to investigate changes in immunological parameters induced by weaning stress (including milk restriction) in calves. Fifteen Holstein calves were subjected to weaning at 6 weeks of age. Blood samples were collected at -14, -7, -2, 1, 3, and 5 days post-weaning (DPW; 0 DPW = 42 days). Weaning caused significant (p < 0.01) increases in the neutrophil (NE):lymphocyte (LY) ratio at 5 DPW with a significant (p < 0.05) reduction of LYs. The concentration of acute-phase proteins (haptoglobin and serum amyloid A) also increased significantly (p < 0.05) at 3 and 5 DPW compared to -2 DPW. Levels of the iron-binding protein lactoferrin decreased significantly (p < 0.05) after weaning. Serum tumor necrosis factor-α and cortisol levels were elevated (p < 0.05) at 3 DPW, while those of serum interferon-γ decreased (p < 0.05) at 1 and 3 DPW compared to levels observed before weaning. Weaning significantly (p < 0.05) decreased the percentage of CD25⁺ T cells in the peripheral blood. In conclusion, weaning stress affected the NE:LY ratio along with the levels of acute phase proteins, lactoferrin, cortisol, and inflammatory cytokines in the peripheral blood of calves. Weaning stress may induce an acute phase response possibly through the elevation of cortisol production and modulation of inflammatory cytokines.     

Copper toxicity in tropical barnacles,Balanus Amphitrite amphitrite and Balanus tintinnabulum tintinnabulum

The effect of Cu on survival time, O₂ consumption and accumulation of Cu in barnacles, Balanus amphitrite amphitrite and Balanus tintinnabulum tintinnabulum was studied. B. amphitrite amphitrite were found to survive longer than B. tintinnabulum tintinnabulum at all concentrations of Cu. Copper reduced the O₂ consumption of both barnacles, but the reduction was greater with B. tintinnabulum tintinnabulum. Copper accumulation in soft parts of the barnacles increased with increasing Cu concentration and time. The barnacles accumulated Cu up to a particular limit (166.7 ± 8.98 μg g^?1 in B. amphitrite amphitrite and 173.6 ± 12.17 μg g^?1 in B. tintinnabulum tintinnabulum) at all Cu concentrations beyond which they died.     

High Performance Thin Layer Chromatography (HPTLC) profile of an insecticidal compound of <Emphasis Type="Italic">Trochus radiatus</Emphasis> and assessment of mucus-associated bacterial extracts

The current study is based on the screening of novel insecticides from new sources that have not being exploited hitherto. The major objectives of this research work were to extract marine molluscs, Lambis lambis, Trochus radiatus and Chicoreus ramosus, from Tuticorin coastal waters using different solvents to test their insecticidal properties and partially purify the active components. The ethyl acetate extracts of L. lambis and T. radiatus showed 100% mortality of Sitophilus oryzae, at a concentration of 200 μg ml⁻¹. The LC₅₀ values for ethyl acetate extracts of L. lambis, T. radiatus and C. ramosus were found to be 67.08, 348.86 and 571.42 μg ml⁻¹, respectively. With regard to bacterial metabolites, at a concentration of 200 μg ml⁻¹ the ethyl acetate extract of A3 was able to elicit an activity of 40% and that of strain A1 – 20%. The LC₅₀ values of the bacterial metabolites were also investigated. Upon chromatographic separation of active ethyl acetate extracts of T. radiatus, the 100% methanol column-purified fraction was found to have an activity of 30% at a concentration of 10 μg against S. oryzae. The purity of the partially pure active compound was observed to be 72.78% on analysis with high pressure thin layer chromatography.     

Plant regeneration from encapsulated nodal segments of Sterculia urens roxb., an endangered gum-yielding tree

The present study is an attempt to prepare synthetic seeds using alginate encapsulation of nodal explants of Sterculia urens. Regeneration potential of encapsulated nodal segments was tested on MS basal, MS + BA (3.0 mg L^?1), MS + TDZ (0.2 mg L^?1), and MS + BA (0.5 mg L^?1) + NAA (2.0 mg L^?1). The regeneration potential was maximum (73.33 ± 1.33) on MS + TDZ (0.2 mg L^?1) followed by (69.33 ± 1.76) on MS + BA (3 mg L^?1) even after 6 months of storage at 4°C, whereas controls did not show any regeneration potential after 1 week. These findings suggest synthetic seed technology as an alternative method for micropropagation and germplasm conservation of Sterculia urens an endangered species.