A method for estimating radioactive cesium concentrations in cattle blood using urine samples

In the region contaminated by the Fukushima nuclear accident, radioactive contamination of live cattle should be checked before slaughter. In this study, we establish a precise method for estimating radioactive cesium concentrations in cattle blood using urine samples. Blood and urine samples were collected from a total of 71 cattle on two farms in the ‘difficult‐to‐return zone’. Urine ¹³⁷Cs, specific gravity, electrical conductivity, pH , sodium, potassium, calcium, and creatinine were measured and various estimation methods for blood ¹³⁷Cs were tested. The average error rate of the estimation was 54.2% without correction. Correcting for urine creatinine, specific gravity, electrical conductivity, or potassium improved the precision of the estimation. Correcting for specific gravity using the following formula gave the most precise estimate (average error rate = 16.9%): [blood ¹³⁷Cs] = [urinary ¹³⁷Cs]/([specific gravity] ? 1)/329. Urine samples are faster to measure than blood samples because urine can be obtained in larger quantities and has a higher ¹³⁷Cs concentration than blood. These advantages of urine and the estimation precision demonstrated in our study, indicate that estimation of blood ¹³⁷Cs using urine samples is a practical means of monitoring radioactive contamination in live cattle.     

Prevalence of extended-spectrum β-lactamase-producing Escherichia coli and Klebsiella pneumoniae in food-producing animals

To evaluate the diversity of extended-spectrum β-lactamases (ESBL) genes among food-producing animals, 48 isolates of ESBL-producing Escherichia coli isolates were obtained from rectal samples of broilers, layers, beef cattle and pigs, at the slaughterhouse level. ESBL-carrying E. coli were isolated from 60.0% of individual broiler rectal samples, 5.9% of layers, 12.5% of beef cattle and 3% of pigs. One ESBL-producing Klebsiella pneumoniae was isolated from a broiler. The ESBL-positive E. coli isolates from broilers harbored various ESBL genes: bla (SHV-12), bla(CTX-M-2), bla(CTX-M-14), bla(CTX-M-15) and bla(CTX-M-44). The plasmid DNAs were analyzed by restriction patterns. Homogeneous band patterns were yielded in those of K. pneumoniae and E. coli isolates harboring the bla(CTX-M-2) gene from different farms. No genetic relation between the 2 CTX-M-14 ESBL-producing strains was found by pulsed-field gel electrophoresis, although 2 plasmids in these strains, obtained from different broiler farms, were similar to each other. This study provides evidence that the proliferation of CTX-M-producing E. coli is due to the growth of indigenous CTX-M-producing strains and the possible emergence of strains that acquired CTX-M genes by horizontal transfer in different broiler farms. CTX-M-producing coliforms in broilers should be controlled due to the critical importance of cephalosporins and the zoonotic potential of ESBL-producing bacteria.     

Ratio of rice reflectance for estimating leaf blast severity with a multispectral radiometer

 Rice reflectance was measured to determine the spectral regions most sensitive to leaf blast infection with a multispectral radiometer. As disease severity increased, reflectance also increased in the 400–500 nm (blue), 570–700 nm (red), and 900–2000 nm regions but decreased in the 500–570 nm and 700–900 nm regions. The increased reflectance in the blue and red regions may be attributed to decreased chlorophyll and carotenoid contents in response to the blast infection. The maximum and minimum reflectance differences occurred at 680 nm and 760 nm for the nondiseased and diseased rice, respectively. The spectral location of maximum sensitivity was 675 nm regardless of disease severity. Rice reflectance ratios were evaluated as indicators of leaf blast severity. Two ratios, R550/R675 (reflectance at 550 nm divided by reflectance at 675 nm), and R570/R675 quantified the significant disease severity. These wavelengths were selected based on the sensitivity minima and maxima. The ratios of nondiseased rice plants varied depending on growth stage. The variation in ratios must be considered when they are used to estimate leaf blast severity. Received: April 2, 2002 / Accepted: August 12, 2002     

Characteristics of Monomeric Al Concentration in An Acidified River

The Kusurisambetsu River is a tributary of the Noboribetsu River. These rivers flow through Hokkaido Prefecture, Japan. Because the upper reaches of the river are acidified by a caldera lake and basin, the river water contains aluminum in high concentration, together with iron and sulfate. The drainage area at the lower reaches is not acidified. Therefore, the acidified water of the river is neutralized by the alkalinity of water from the drainage area. The water at the mouth is neutral in pH, although about 1 mg/L of aluminum is detected. Heavy rainfall occurred in Autumn 1993, and many adult salmon were killed at the lower reaches of the river. This prompted study of the neutralization process of river water, the Al concentration change, and the influence of rainfall. Determination of Al found total Al concentration to be roughly constant at the mouth of the Noboribetsu River. However, the concentration of monomeric Al varied with pH, which varied according to rainfall and flow volume. Monomeric Al showed a minimum concentration of roughly pH 6.0. At pH of greater than 6.0, the presence of some kinds of polymeric Al is determined by filtration fraction. This suggests that the death of salmon is caused by monomeric Al when conditions are acidic. Liming test of the river water confirmed that the monomeric Al that was toxic under acidic conditions became non-toxic under alkaline conditions.     

Studies on the bacteria isolated anaerobically from paddy field soil III. Production of fatty acids and ammonia by Clostridium species

The volatile fatty acids produced by the representative strains of clostridia isolated from paddy field soil were surveyed. All the strains, except Cl. tertium, utilized only amino acids as the sole source of energy for growth. All of them produced acetic and butyric acids, and some of them produced propionic acid in addition. Iso-valeric and iso-butyric acids were also produced, except by Cl. tertium. These results suggest that all the strains, except Cl. tertium, may carry out the STICKLAND reaction. The cells of each strain harvested from the culture in the VL medium were subjected to examination whether or not they carried out the STICKLAND reaction in the presence of leucine or glycine. All the strains, except Cl. tertium, catalyzed the STICKLAND reaction and some strains also metabolized leucine and glycine even when they were supplied singly.     

A passage and storage system for isolated bovine endometrial epithelial and stromal cells

To establish a storage system for isolated endometrial cells, we investigated the basal, oxytocin (OT)- and tumor necrosis factor (TNF) alpha-stimulated production of prostaglandin (PG) F(2alpha) in bovine-passaged and frozen-thawed endometrial cells. Stromal and epithelial cells obtained from cows in the early stage of the estrous cycle (Days 2-5) were frozen at -80 C or further cultured and/or passaged until passage 4 in DMEM/Ham's F-12 supplemented with 10% calf serum. A fresh-unfrozen primary culture and one-time passaged fresh-unfrozen cells were used as the control. When both unfrozen and frozen cells reached confluence, the culture medium was replaced with fresh medium with 0.1% BSA and the cells were stimulated with OT (100 ng/ml) or TNFalpha (1 ng/ml) for 4 h. The passage and freezing of the endometrial cells did not affect their morphology. In primary culture of frozen and unfrozen endometrial cells, OT strongly stimulated PGF(2alpha) production in epithelial cells, and TNFalpha strongly stimulated PGF(2alpha) production in stromal cells (P<0.05). The basal output of PGF(2alpha) in frozen stromal cells was similar to that in unfrozen stromal cells. However, the basal output of PGF(2alpha) in frozen epithelial cells was significantly lower than that unfrozen cells (P<0.05). On the other hand, in passaged cells, the basal level of PGF(2alpha) production was retained until passage 1 in epithelial cells, whereas it was retained until passage 4 in stromal cells. Although epithelial cells responded to OT in PGF(2alpha) production until passage 2 (P<0.05), the stromal cells showed a significant response to TNFalpha until passage 4 (P<0.05). These results suggest that stored cells could be used for studying the physiology of bovine endometrium in vitro until passage 1 in endometrial epithelial cells, and until passage 4 in stromal cells.     

Breed differences in genotype and allele frequency of catechol O-methyltransferase gene polymorphic regions in dogs

Catechol O-methyltransferase (COMT) inactivates catecholamines and catechol-containing drugs such as L-DOPA. The common genetic polymorphism Val158Met in the human COMT gene is suspected to be associated with "persistence" or risk for schizophrenia. In this study, we attempted to identify the canine COMT gene fragment and to find a similar polymorphism and to reveal its genetic distribution among five representative canine breeds. We found that the amplified gene consisted of 663 bp nucleotides and was 84% homologous with the human COMT gene. The single nucleotide polymorphisms, guanine adenine substitution, were observed at the 39th, 216th and 482nd nucleotides. From the genotyping of the 216th polymorphism among 266 dogs by the polymerase chain reaction-restriction fragment length polymorphism method with restriction enzyme EagI, and that of the 482nd polymorphism with restriction enzyme SfcI, we found inter-breed variations of genotypes as well as of allelic frequencies for both of these polymorphic regions. These results suggest that the identified polymorphisms will be useful tools in elucidating the genetic background of canine behavioral traits.     

Wind‐induced stock variation of the neon flying squid (Ommastrephes bartramii) winter–spring cohort in the subtropical North Pacific Ocean

Our examination of the neon flying squid (Ommastrephes bartramii) winter–spring cohort catch per unit effort (CPUE, an index of stock) revealed significant positive correlations with the interannual variations of observed chlorophyll‐a (Chl‐a) concentration and autumn–winter mixed layer depth (MLD) in the winter–spring feeding grounds of paralarvae and juveniles (130–170°E, 20–27°N). These correlations suggest the importance of integrated bottom‐up effects by the autumn–winter MLD for the neon flying squid stocks. However, the influence of autumn–winter MLD interannual variation in the forage availability for paralarvae and juveniles, i.e., particulate organic matter and zooplankton, has still been unclear. In this study, we use the lower trophic ecosystem model NEMURO, which uses the physical environmental data from the ocean reanalysis dataset obtained by the four‐dimensional variational (4DVAR) data assimilation method. The model‐based investigation enables us to clarify how the autumn–winter MLD controls the particulate organic matter and zooplankton abundance in the feeding grounds. Further, our investigation of the autumn–winter MLD interannual variation demonstrates that the stronger autumn wind in the feeding grounds develops a deeper mixed layer. Therefore, the deep mixed layer entrains nutrient‐rich water and enhances photosynthesis, which results in good feeding conditions for paralarvae and juveniles. Our results underline that the wind system interannual variation has critical roles on the winter–spring cohort of the neon flying squid stock.