Transfection of non-susceptible cells with Ovis aries recombinant lymphocyte function-associated antigen 1 renders susceptibility to Mannheimia haemolytica leukotoxin

Mannheimia haemolytica is an important etiological agent of pneumonia in domestic sheep (DS, Ovis aries). Leukotoxin (Lkt) produced by this organism is the principal virulence factor responsible for the acute inflammation and lung injury characteristic of M. haemolytica caused disease. Previously, we have shown that the leukocyte-specific integrins, beta(2) integrins, serve as the receptor for Lkt. Although it is certain that CD18, the beta subunit of beta(2) integrins, mediates Lkt-induced cytolysis of leukocytes, it is not clear whether CD18 of all three beta(2) integrins, LFA-1, Mac-1 and CR4, mediates Lkt-induced cytolysis of DS leukocytes. Since polymorphonuclear leukocytes, which express all three beta(2) integrins, are the leukocyte subset that is most susceptible to Lkt, we hypothesized that all three beta(2) integrins serve as the receptor for Lkt. The objective of this study was to determine whether DS LFA-1 serves as a receptor for M. haemolytica Lkt. We cloned the cDNA for DS CD11a, the alpha subunit of LFA-1, and co-transfected it along with the previously cloned cDNA for DS CD18, into a Lkt-non-suceptible cell line. Transfectants stably expressing DS LFA-1 were bound by Lkt. More importantly, Lkt lysed the DS LFA-1 transfectants in a concentration-dependent manner. Pre-incubation of Lkt with a Lkt-neutralizing monoclonal antibody (MAb), or pre-incubation of transfectants with MAbs specific for DS CD11a or CD18, inhibited Lkt-induced cytolysis of the transfectants. Exposure of LFA-1 transfectants to low concentrations of Lkt resulted in elevation of intracellular [Ca(2+)](i). Taken together, these results indicate that DS LFA-1 serves as a receptor for M. haemolytica Lkt.     

The generation and persistence of genetic variation in foot-and-mouth disease virus

Genetic variation in foot-and-mouth disease virus (FMDV) is of interest for at least two reasons. First, changes to the genes encoding capsid proteins results in antigenic variation, and affects vaccine efficiency and effectiveness of vaccination programs; second, genetic changes can lead to important insights into the transport of virus between countries, regions, herds, and even possibly individuals. Current estimates of RNA virus mutation rates suggest that an average of about one base mis-incorporation is likely to occur each time a single FMDV genome replicates. This should result in the introduction of every possible 1-step mutation from the progenitor genotype into the viraemia of a single infected animal many times a day. In the absence of purifying selection, a single infected animal should therefore generate a genetically very diverse population of virus.Viral-capsid sequences obtained from infected animals sampled over long-term FMDV epidemics suggest that these genetic changes accrue in a remarkably linear 'clock-like' fashion and at rates of around 1% change per year. While such a rate is generally regarded as quite high, it is actually somewhat lower than one might expect based on the rate at which viral diversity could be generated within a single animal. The difference might be explained in a variety of possible ways: (1) the mutation rate has been overestimated; (2) purifying selection is stronger than predicted; (3) only a restricted subset of excreted virus is actually infectious; (4) infected animals only excrete virus from a small partitioned subset of amplified virus, and that most of the generated viral diversity is unable to exit the animal; or (5) only a small fraction of all infected animals participate in the actual disease-transmission process.     

Mechanisms conferring resistance of Western flower thrips to bendiocarb

Mechanisms associated with bendiocarb resistance were examined in two strains of western flower thrips, Frankliniella occidentalis (Pergande), that differed in their susceptibility to this carbamate by 13.6-fold. No appreciable differences between the two strains in [¹⁴C] bendiocarb penetration and excretion were detected; however, bendiocarb was metabolised substantially faster by the resistant KCM thrips than by the more susceptible UMC thrips. No appreciable difference between the two strains was found in the sensitivity of acetylcholinesterase activity to inhibition by bendiocarb. It was concluded that bendiocarb resistance in KCM western flower thrips was due to enhanced metabolism that probably was mainly oxidative in nature.     

An investigation on commercial farms of factors thought to contribute to egg cracking

An investigation of 52 egg production units showed that in battery units, but not under other systems of husbandry, the proportion of defective eggs produced by birds in their second year was more than twice that of the first year. The proportion of defective eggs was unrelated to the shell thickness (measured by the deformation test on random samples of sound eggs from the same flocks). Damage to eggs during collection and packing was trivial, relative to the proportions of defective eggs found in the nests or cages.     

Oxygen inhibition of acetylene reduction (nitrogen fixation) in soil: Effect of glucose and oxygen concentrations

Sandy loam soil was amended with different concentrations of glucose and was incubated at different pO₂ levels. Under many conditions of incubation time and treatment, N₂ ase activity as determined by 1-h aerobic C₂H₂ reduction assay (flushed with Ar:O₂, 4:1 before assay) was significantly less than that determined by means of ambient assay (carried out at the pO₂ of incubation without flushing with Ar:O₂, 4:1 before assay). The difference between the N₂ase activity in aerobic assay and that in ambient assay increased with decreasing glucose and O₂ concentrations imposed during incubation. The inhition in aerobic assays was analogous to O₂-induced shut-off of N₂ase and amounted to 75 per cent inhibition after incubation at 0.06 atm pO₂ of samples amended with 0.75% glucose (w/w). Similar O₂ inhibition was observed after amendment with mannitol and with lactate. Times of incubation were chosen such that development of anaerobic N₂ase activity was either absent or too low to account for the observed effects of O₂ during assay. It was shown that 0.05 atm pC₂H₂ was adequate for routine 1-h assays of the soil system employed. Individual soil samples could be subjected to repeated 1-h assays (with removal of C₂H₂ and C₂H₄ by evacuation after each assay) thus avoiding side-effects of long exposure to C₂H₂.     

Effect of electrical stunning at slaughter on the quality of farmed turbot (Psetta maxima)

Food quality aspects of farmed turbot (Psetta maxima) were compared following two methods of slaughter: the current commercial method, by immersion in an ice slurry, which is then dewatered after approximately 20 min, or by first humanely, electrically stunning the fish using a prototype commercial stunner, before immersion in an ice slurry, which is dewatered after 20 min. Quality was assessed for up to 10 days of storage on ice following slaughter. No differences were found between the slaughter methods in terms of an overall carcass quality: overall appearance, haemorrhage, damage, burst gall bladder, staining of the body cavity by leakage from the gut or damage to the spine. No detectable difference was found between the treatments using the industry standard freshness scoring system, the Quality Index Method. Both groups of fish were classified as ‘Fresh’ after 10 days of storage on ice. Using objective measurements of colour, no differences between fish from either treatment were found in fillet colour. Changes in flesh pH were similar in electrically stunned and traditionally killed fish with a mean pH (±SE) at 2 h post‐mortem of 6.80±0.027 declining to 6.44±0.032 at 24 h post‐mortem. Humane electrical stunning of turbot at slaughter neither detectably improved nor decreased product quality as measured between 1 and 10 days of storage on ice.