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.     

Impact of High Night‐Time and High Daytime Temperature Stress on Winter Wheat

High temperature is a major environmental factor that limits wheat (Triticum aestivum L.) productivity. Climate models predict greater increases in night‐time temperature than in daytime temperature. The objective of this research was to compare the effects of high daytime and high night‐time temperatures during anthesis on physiological (chlorophyll fluorescence, chlorophyll concentration, leaf level photosynthesis, and membrane damage), biochemical (reactive oxygen species (ROS) concentration and antioxidant capacity in leaves), growth and yield traits of wheat genotypes. Winter wheat genotypes (Ventnor and Karl 92) were grown at optimum temperatures (25/15 °C, maximum/minimum) until the onset of anthesis. Thereafter, plants were exposed to high night‐time (HN, 25/24 °C), high daytime (HD, 35/15 °C), high daytime and night‐time (HDN, 35/24 °C) or optimum temperatures for 7 days. Compared with optimum temperature, HN, HD and HDN increased ROS concentration and membrane damage and decreased antioxidant capacity, photochemical efficiency, leaf level photosynthesis, seed set, grain number and grain yield per spike. Impact of HN and HD was similar on all traits. Greater impact on seed set, grain number and grain yield per spike was observed at HDN compared with HN and HD. These results suggest that HN and HD during anthesis cause damage of a similar magnitude to winter wheat.     

Variability in concentrations of zinc in serum and feed when using zinc oxide as a supplement for the prevention of facial eczema

AIMS: To determine the variability of concentrations of Zn in feed, when used as a supplement to prevent facial eczema, and to determine the variability in concentrations of Zn in serum between cows and herds that are being supplemented with ZnO in feed, using in-shed feeders or on a feed pad.

METHODS: Sixteen commercial dairy farms in the Waikato region of New Zealand were enrolled, that were supplementing cows with ZnO in the feed using either an automatic in-shed feeder (ASF) or a feed pad (FP) using a feed-out or mixer wagon. On each farm 10 cows were selected by the farmer, that were assumed to be representative of the age and liveweight of the herd. Four hours after supplement feeding, each cow was weighed and a blood sample collected for measurement of concentrations of Zn in serum. Three samples of feed were collected from each farm for Zn analysis, from the beginning, middle and end of the feed being distributed. Levene’s test for homoscedasticity was used to analyse whether there were differences in variation of individual concentrations of Zn in serum, and in the feed, between the two feeding systems. Multivariable linear regression was used to examine associations between age, feeding method or liveweight and concentrations of Zn in serum, after accounting for the variability between farms.

RESULTS: Of the 163 cows sampled, concentrations of Zn in serum were between 20–35?µmol/L in 75/163 (46 (95% CI=38–54)%) cows; were <20?µmol/L in 71/163 (44 (95% CI=36–52)%) cows, and >35?µmol/L in 17/163 (10 (95% CI=6–16)%) cows. The variation in concentrations of Zn in serum in individual cows differed between farms (p<0.001), and the variability was greater for cows fed using a FP than ASF (p<0.001). There was no difference in the variation of concentrations of Zn in feed between the two feeding methods (p=0.54), but concentrations of Zn in serum were associated with the amount of Zn offered in feed (p=0.008).

CONCLUSIONS AND CLINICIAL RELEVENCE: There was significant variability between farms in the concentrations of Zn in the serum of cows being supplemented with ZnO in feed. Only 46% of cows sampled had concentrations of Zn between 20–35?µmol/L. Effective management of facial eczema should include monitoring Zn in the feed and in serum to ensure cows are receiving the correct dose they require.     

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.     

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.     

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.     

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.