Pasteurella haemolytica A1 and Bovine Respiratory Disease: Pathogenesis

The severe fibrinonecrotic pneumonia associated with pneumonic pasteurellosis usually results from colonization of the lower respiratory tract by Pasteurella haemolytica biotype A, serotype 1(A1). Despite recent research efforts, the authors lack a detailed understanding of the interactions and host response to P. haemolytica in the respiratory tract. The authors hypothesize that management and environmental stress factors or viral infection alters the upper respiratory tract (URT) epithelium allowing P. haemolytica to colonize the epithelium. Once the URT is colonized, large numbers of organisms enter the lung where they interact with alveolar macrophages. Endotoxin, released from the bacteria, crosses the alveolar wall where it activates pulmonary intravascular macrophages, endothelium, neutrophils, lymphocytes, platelets, complement, and Hageman factor leading to complex interactions of cells and mediators. It is the progression of this inflammatory response with neutrophil influx that is ultimately responsible for the pulmonary injury. Leukotoxin is a major virulence factor of P. haemolytica that allows it to survive by destroying phagocytic cells. At subcytolytic concentrations it may also enhance the inflammatory response by activating cells to produce mediators and release reactive oxygen metabolites and proteases.     

Effects of Pasteurella haemolytica A1 leukotoxin on bovine neutrophils: degranulation and generation of oxygen-derived free radicals.

To further define the role of Pasteurella haemolytica A1 leukotoxin in the pathogenesis of bovine pneumonic pasteurellosis, its in vitro effects on bovine neutrophils were investigated. Leukotoxin-containing culture supernatant, from P. haemolytica, stimulated a neutrophil respiratory burst as measured by the generation of oxygen-derived free radicals O2- and H2O2. This effect was immediate because preincubation of neutrophils with the culture supernatant for 5 min or longer substantially suppressed this respiratory burst. This suppression was due to cytolysis of the neutrophils. Prolonged incubation of neutrophils with the same culture supernatant caused further cytolysis and degranulation. Heat-inactivated P. haemolytica culture supernatant that had lost its cytotoxic properties failed to stimulate respiratory burst by neutrophils. Furthermore, the respiratory burst, cytolysis and degranulation were abrogated only by leukotoxin-neutralizing monoclonal and polyclonal antibodies, but not by antibodies against the lipopolysaccharide. These studies show that the leukotoxin component in the culture supernatant was responsible for the generation of oxygen-derived free radicals and proteolytic enzymes from neutrophils which may participate in direct lung injury.     

Production of polyclonal antiserum against recombinant VP28 protein and its application for the detection of white spot syndrome virus in crustaceans

The VP28 gene of white spot syndrome virus (WSSV) was cloned into pRSET B expression vector. The VP28 protein was expressed as a protein with a 6-histidine taq in Escherichia coli GJ1158 with NaCl induction. Antiserum was raised against this recombinant-VP28 protein in rabbits and it recognized VP28 protein in naturally and experimentally WSSV-infected shrimp, marine crabs, freshwater prawns and freshwater crabs. The antiserum did not recognize any of the other known WSSV structural proteins. Various organs such as eyestalks, head muscle, gill tissue, heart tissue, haemolymph, tail tissue and appendages were found to be good materials for detection of WSSV using the antiserum and detection of WSSV was successful in experimentally infected Penaeus monodon and P. indicus at 12 and 24 h post-infection (p.i.), respectively. The antiserum was capable of detecting WSSV in 5 ng of total haemolymph protein from WSSV-infected shrimp.     

Microsatellite Marker Based Assessment of Genetic Diversity among Cultivars, Landraces and Wild Relatives in Rice

India being the primary center of origin for rice had a very large treasure of local land races, most of which are out of cultivation today. The exact genetic potential and their differences from commercial varieties and the magnitude of heterogeneity still present in them are not well catalogued. Hence the need to characterize the available land races has become imminent in the modem day concept of crop improvement （Rezai and Frey, 1990）. The present study addresses the utility of SSR markers in divulging the genetic relationships at molecular level among the major component factions office gennplasm viz., local cultivars, land races and wild species collected from a wide range of agro-geographical regions of Indian subcontinent.     

Mortality of the tobacco caterpillar,Spodoptera litura (F.), when treated withBacillus Thuringiensis combinations with boric acid and insecticides

Bacillus thuringiensis Berliner in combination with 0.1, 0.5, 1.0, 1.5 and 2% boric acid caused 100% mortality of the tobacco caterpillar,Spodoptera litura (F.), irrespective of instar, in a much shorter period (1.77 to 3.61 days) than with boric acid or the pathogen alone. The toxicity ofB. thuringiensis in combination with dicrotophos at 0.02 and 0.04%, with fenitrothion at 0.025 and 0.05%, or with dichlorvos at 0.01 and 0.05%, was enhanced.     

Influence of packaging method and storage time on shear value and mechanical strength of intramuscular connective tissue of chevon

The objectives of this study were to determine the effects of storage time (ST) and packaging method (PM) on tenderness and changes in intramuscular connective tissue (IMCT) strength of chevon. Spanish does (8 mo of age, average BW 25 kg) were harvested (n = 12), chilled at 4 degrees C for 24 h, and then fabricated into 2.5-cm-thick leg, shoulder/arm, and loin/rib cuts. The cuts from six carcasses were vacuum-packed and aged at 2 degrees C for 0, 4, 8, or 12 d. To assess the influence of a packaging method that favors oxidation on postmortem tenderization, the cuts from the remaining six carcasses were placed on styrofoam trays, overwrapped with polyvinyl-chloride film, and stored at 2 degrees C for similar periods. At each ST, longissimus (LM), semimembranosus (SM), and triceps brachii (TB) muscles were assessed for Warner-Bratzler shear (WBS) values. The WBS of uncooked meat, myofibrillar fragmentation index (MFI), and collagen solubility were assessed on LM. The IMCT samples were prepared to assess changes in mechanical strengths and for scanning electron microscopy (SEM). Intact honeycomb structures of endomysium, with no muscle fiber elements, were observable under SEM. The PM or ST did not influence the mechanical strength of IMCT preparations, as measured by a texture analyzer. Collagen solubility of LM muscles also did not change during aging. For both PM, cooked meat WBS values were higher (P < 0.01) in SM and TB than in LM. In the SM samples, the average WBS values were higher (P < 0.01) at d 0 than at other ST. Although MFI of LM increased with increasing aging time (P < 0.05), changes in WBS over ST were minimal in TB and LM samples. The WBS of uncooked LM decreased sharply up to 8 d postmortem in both PM (P < 0.05). However, there was no PM x ST interaction to indicate any adverse influence of packaging on tenderization of chevon. The results suggest that aging chevon cuts for more than 4 d may not result in significant additional improvement in tenderness.     

Effect of dietary protein on intake,nutrients utilization,nitrogen balance,blood metabolites,growth and puberty in growing Bhadawari buffalo (Bubalus bubalis) heifers

Fifteen Bhadawari buffalo heifers of 207?±?9.78 kg mean body weight were randomly distributed into three dietary groups to evaluate the effect of protein level on nutrient utilization, nitrogen (N) balance, growth rate, blood metabolites, and puberty. All animals were offered wheat straw-berseem diets supplemented with concentrate mixtures of similar energy (2.7 Mcal/kg) and different protein levels (14.3–22 %). Animals of standard-protein group (SPG) were offered protein and energy as per requirement, while animals of low-protein group (LPG) and high-protein group (HPG) were fed 20 % less and 20 % more protein, respectively, than SPG. Feed dry matter (DM) and metabolizable energy (ME) intake (% body wt. and g/kg w^0.75) were similar for all three diets; however, the crude protein (CP) and digestible crude protein (DCP) intake on percent body weight and per kilogram metabolic weight was higher (P?<?0.05) in HPG than in SPG or LPG. Digestibility of CP, cellulose, and hemicellulose was higher (P?<?0.05) in HPG versus LPG. Fecal N excretion was similar, while urinary N excretion was highest (P?<?0.05) in HPG (74.83 g/day) compared with SPG (50.03 g/day) and LPG (47.88 g/day), which resulted in lower N retention in HPG than in the other dietary groups. Level of dietary N had no effect on blood metabolites viz. glucose, urea, and N. Digestible energy (DE) and ME contents of diets were identical, while DCP contents were higher (P?<?0.05) in HPG than in LPG. Feed and nutrient (CP and ME) conversion efficiency to produce a unit kilogram weight gain was identical among the dietary groups. Dietary protein level had no effect on the heifer’s weight and age at puberty. The mean growth rate of heifers at 240 days was higher (P?>?0.05) in SPG (330.8 g/day) than in LPG (296.7 g/day), while the animals gained more weight in January to March months and the lowest weight in May to July months. Protein level had no effect on conception rate of heifers. Results revealed that 20 % higher or less protein than the ICAR requirement had no significant (P?>?0.05) on feed intake, nutrient conversion efficiency for weight gain, heifer growth, and puberty; however, 20 % more protein increased urinary N loss.

     

Estimation of Deposition Velocities for 85Sr, 131I, 137Cs on Spinach,Radish and Beans Leaves in a Tropical Region Under Simulated Fallout Conditions

Poly dispersive aerosols of SrCl₂, NaI, CsCl having a size distribution of 2.33 m (Activity Median Aerodynamic Diameter, [AMAD]) with a geometric standard deviation of 1.83 m are used for the determination of deposition velocity for ⁸⁵Sr, ¹³¹I and ¹³⁷Cs aerosols on tropical spinach (Spinicia olericia), radish (Raphanous sative) and beans (Phasolous valugeries) plant leaves. The experiments were carried out in a specially designed exposure chamber to simulate deposition under accidental releases of radioactivity in tropical environment. The rates of particles deposition are expressed as a function of plant surface area and of plant dry weight. The deposition velocities obtained are in the range of 10^–6 to 10^–7 m/s. These values differ significantly from those in temperate regions. The lower values for deposition velocities (v _g) for the tropical environment is attributed due to high humidity. Therefore, the deposition of suspended aerosols on leaf surfaces caused by impaction and Brownian diffusion becomes slower in tropical environment and resulted into lower value of deposition velocities.     

Specific mutations induced by triplex-forming oligonucleotides in mice

Triplex-forming oligonucleotides (TFOs) recognize and bind to specific duplex DNA sequences and have been used extensively to modify gene function in cells. Although germ line mutations can be incorporated by means of embryonic stem cell technology, little progress has been made toward introducing mutations in somatic cells of living organisms. Here we demonstrate that TFOs can induce mutations at specific genomic sites in somatic cells of adult mice. Mutation detection was facilitated by the use of transgenic mice bearing chromosomal copies of the supF and cII reporter genes. Mice treated with a supF-targeted TFO displayed about fivefold greater mutation frequencies in the supF gene compared with mice treated with a scrambled sequence control oligomer. No mutagenesis was detected in the control gene (cII) with either oligonucleotide. These results demonstrate that site-specific, TFO-directed genome modification can be accomplished in intact animals.     

Antibodies to the core protein of lymphadenopathy-associated virus (LAV) in patients with AIDS

Lymphadenopathy-associated virus ( LAV ), a human T- lymphotrophic retrovirus isolated from a homosexual man with lymphadenopathy, has been causally associated with acquired immunodeficiency syndrome (AIDS). A sensitive and specific radioimmunoprecipitation test was developed for the detection of antibodies to the major core protein of LAV , p25 (molecular weight 25,000). Antibody to LAV p25 was found in the serum of 51 of 125 AIDS patients, 81 of 113 patients with lymphadenopathy syndrome, 0 of 70 workers at the Centers for Disease Control (some of whom had handled specimens from AIDS patients), and 0 of 189 random blood donors. Of a group of 100 homosexual men from San Francisco whose serum was obtained in 1978, only one had antibody to LAV p25; in contrast, of a group of 50 homosexual men in the same community whose serum was obtained in 1984, 12 had antibodies to LAV p25.