Mechanism of neuromuscular block by chlordimeform

The mechanism of action of chlordimeform on the frog sciatic nerve-sartorius muscle preparation has been studied by means of microelectrodes. Chlordimeform (0.1 mM) suppresses the amplitude of spontaneous miniature end-plate potentials without much changing their frequency. At 1 mM, it completely blocks the miniature end-plate potentials. The resting membrane potential is not affected. The end-plate potential evoked by nerve stimulation is also suppressed, while the action potential from the nerve terminal is not impaired. The sensitivity of the end-plate membrane to the transmitter substance as examined by iontophoretic applications of acetylcholine is effectively decreased by chlordimeform. The quantal content of transmitter release is not affected. It is concluded that the block of neuromuscular transmission by chlordimeform is due primarily to a depression of the end-plate sensitivity to the transmitter.     

Quantitative structure-activity study of herbicidal N-aryl-3,4,5,6-tetrahydrophthalimides and related cyclic imides

The quantitative relationship between structure and inhibitory activity against the root growth of sawa millet of 30 N-aryl-3,4,5,6-tetrahydrophthalimides and 88 related N-aryl cyclic imide compounds (3-aryl-1,5-tetramethylenehydantoins, 4-aryl-1,2-tetramethylenetriazolidine-3,5-diones, and their thiocarbonyl analogs) was analyzed by means of physicochemical parameters of the aryl substituents and regression analysis. The results showed that, in each series of these classes of compounds, the position-specific steric effects of aromatic substituents, as represented by Verloop's STERIMOL values, are highly important in determining the inhibitory potency of the compound.     

Development of furnished cages re‐using conventional cages for laying hens: Behaviour,physical condition and productivity

Furnished cages for laying hens have advantages in allowing normal behaviors and maintaining productivity. As the cost of introduction is a barrier for farms, we developed furnished cages that re‐use conventional cages. To determine the minimum and functional cage design, we compared six designs, combinations of two floor designs (artificial turf or wire cage floor) and three screening designs in the integrated area (no screening, one entrance side or four sides). In total, 144 hens were used, and we measured behavior, physical condition and productivity. Comparing the floors, the percentages of hens performing dust‐bathing and laying eggs in the integrated area were higher in cages with turf than wire floor (P < 0.05 for both). Comparing the screening, dust‐bathing, litter‐exploring and active behavior tended to be more frequent in cages with the integrated area screened on one side than four sides. Feather damage was lower in cages with the integrated area screened on one side than with no screening (P < 0.05). These results suggest that the cage design with an integrated area with artificial turf floor, screened on one side, was effective for furnished cages that re‐use conventional cages.     

Phage conversion of exfoliative toxin A in Staphylococcus aureus isolated from cows with mastitis

An exfoliative toxin produced by Staphylococcus aureus is the causative agent of staphylococcal scalded-skin syndrome (SSSS) in young children. Recently, we reported that only few isolates of S. aureus from bovine mastitis contained the eta gene encoding exfoliative toxin A (ETA) and produced ETA in vitro. In this study, we isolated temperate phages from two ETA-positive bovine isolates of S. aureus by treatment with mitomycin C. Polymerase chain reaction (PCR) assay of the phage genomes suggested that the temperate phages carried the structural gene for ETA. Moreover, the nucleotide sequence analysis of the PCR products revealed that the eta gene was located very close to an amidase gene on the phage genomes. The nucleotide sequence for the amidase gene of the bovine phage (bovine phi ETA) differed at nine positions from that of the amidase gene of phi ETA from a human isolate reported by Yamaguchi et al. [Mol. Microbiol. 38 (2000) 694], suggesting that eta-converting phages are heterogeneous. Bovine phi ETA had a head with a hexagonal outline and a non-contractile and flexible tail. Bovine phi ETA was able to lysogenize ETA-negative bovine isolates of S. aureus, and the lysogenized S. aureus isolates had the ability to produce ETA. These results suggest the possibility of horizontal transmission of the eta gene by temperate bacteriophages among bovine isolates of S. aureus.     

Application of highly differentiated SNPs between Japanese Black and Holstein to a breed assignment test between Japanese Black and F1 (Japanese Black x Holstein) and Holstein

Two taurine breeds, Japanese Black and Holstein, established from geographically distant origins and selected for different uses, beef and dairy, were extensively genotyped using a genome‐wide single nucleotide polymorphism (SNP) chip with more than 1000 animals of each breed. The genetic structure was examined by principal component analysis, in which the first principal component clearly separated the two breeds and explained more than 15% of the variance. Highly differentiated SNPs were detected throughout the genome, some of which were clustered within small regions on BTA4 (79.2–79.7 Mb, Btau4.0) and BTA26 (22.2–23.6 Mb). A breed assignment test was developed using 18 highly differentiated SNPs to distinguish Japanese Black from F₁ (Japanese Black × Holstein) and Holstein. The error rate that an F₁ or Holstein animal is misjudged as Japanese Black was expected to be < 0.8%, while the error rate that a Japanese Black animal is misjudged as F₁ or Holstein was expected to be < 0.001%. This test provides a reliable and powerful method to detect breed label falsification in retail beef.     

Overall welfare assessment of laying hens: Comparing science‐based,environment‐based and animal‐based assessments

To increase the validity of evaluations and facilitate expansion and maintenance of assessment systems, we constructed a database of studies on the welfare of laying hens around the world. On the basis of this database, we devised a science‐based welfare assessment model. Our model includes measurements, levels and weightings based on the scientific studies in the database, and can clarify the advantages and disadvantages of housing systems for laying hens from the viewpoint of the five freedoms. We also evaluated the usefulness of our model by comparing it with environment‐based Animal Needs Index (ANI), another science‐based model called FOWEL, and animal‐based measurements. Our model showed that freedom from injury, pain and disease, and freedom from discomfort were more secure in the cage system, while non‐cage systems scored better for natural behavior and freedom from fear and distress. A significant strong‐positive correlation was found between the animal‐based assessment and the total scores of ANI (rs = 0.94, P < 0.05), FOWEL (rs = 0.99, P < 0.05) or our model (rs = 0.99, P < 0.05), which indicate that these different approaches to welfare assessment may be used almost interchangeably to ‘measure’ a common property (‘overall laying hen welfare’). However, assessments using our model and FOWEL were more sensitive than ANI and can be applied to cage systems, which suggest that our model and FOWEL may have added value.     

Enantioseparation of imazalil residue in orange by capillary electrophoresis with 2-hydroxypropyl-beta-cyclodextrin as a chiral selector

Chiral resolution of imazalil, a fungicide, was performed by capillary electrophoresis (CE) using 2-hydroxypropyl-beta-cyclodextrin as a chiral selector. Factors affecting the chiral resolution and migration time of imazalil were studied. The optimum running conditions were found to be 5 mM ammonium dihydrogenphosphate-50 mM phosphate buffer (pH 3.0) containing 4 mM 2-hydroxypropyl-beta-cyclodextrin with an effective voltage of +25 kV at 20 degrees C using direct detection at 200 nm. Under these conditions, the resolution (Rs) of racemic imazalil was approximately 6. The extraction of imazalil from orange samples was done with acetonitrile under basic conditions. The extract was purified with a solid-phase extraction cartridge (Sep-Pak plus PS-2) and was analyzed by the above CE method. Eight orange samples were analyzed, and imazalil was detected in seven samples. In four of these seven oranges, the level of (-)-imazalil was the same as that of (+)-imazalil, but in the other three oranges, the level of (-)-imazalil was found to be lower than that of (+)-imazalil, suggesting that (-)-imazalil was degraded more quickly than (+)-imazalil in oranges.     

Quantitative structure–activity studies of insect growth regulators: XVI. Substituent effects of dibenzoylhydrazines on the insecticidal activity to Colorado potato beetle Leptinotarsa decemlineata

Insecticidal activity against the Colorado potato beetle, Leptinotarsa decemlineata, was measured for a series of substituted N-tert-butyl-dibenzoylhydrazines, in which one of the benzoyl moieties closer to the tert-butyl group was fixed as being 2-chloro-substitued and the other variously substituted singly or doubly. The effects of substituents on the activity were quantitatively analysed using the classical quantitative structure–activity relationship (QSAR) procedure. The activity against the Colorado potato beetle increases with the molecular hydrophobicity. In addition, various types of steric effect are at work, depending upon the positions. Hydrogen-bonding acceptor substituents at the para position enhance the activity. There seem to be threshold (or optimum) values, albeit position-dependent, in the molecular hydrophobicity, above which the activity starts to decrease. This biphasic contribution of the molecular hydrophobicity to activity against coleopterous larvae is the most conspicuous difference in substituent effects from those found for similar compounds against lepidopterous pest insects, and may be the basis of the variations in the activity spectrum for certain compounds in this series. The introduction of bulkier substituents into the meta- and para-positions of the benzene ring, apart from the tert-butyl group, is unfavorable to activity. LD₅₀ values against Colorado potato beetle larvae of methoxyfenozide (RH-2485) and tebufenozide (RH-5992) were in the order of 10⁻⁷ mol per insect, whereas those of RH-5849, and halofenozide (RH-0345) were very low, 10⁻⁹–10⁻¹⁰ mol per insect being selective to the coleopterous larvae. © 1999 Society of Chemical Industry     

Transgenic Nile tilapia (Oreochromis niloticus) over-expressing growth hormone show reduced ammonia excretion

The growth of aquaculture has negatively affected the environment due to the high levels of nitrogen excreted by farmed fish. Here we propose that modifying the nitrogen metabolism of the fish themselves using transgenic technology might solve the pollution problem. Growth hormone (GH) is known to increase protein retention and absorption, and is thought to reduce ammonia excretion. Thus, we produced transgenic Nile tilapia (Oreochromis niloticus) that over-expressed the GH gene throughout their bodies. Our findings showed that the food-conversion efficiency of the transgenic fish was 35% higher than that of their non-transgenic siblings. The rearing period required for the transgenic fish to reach a body weight of 20 g was about 75% of that required for non-transgenic fish that were fed the same type and quantity of food. The total amount of ammonium-nitrogen excreted by the transgenic fish was about 69% of that excreted by the wild-type fish over their lifetime. These results suggest that our transgenic approach has the potential to reduce the amount of nitrogen pollution caused by farmed fish. This strategy is a promising option for making aquaculture more ‘eco-friendly’.