Loop-mediated isothermal amplification for the rapid, sensitive, and specific detection of the O9 group of Salmonella in chickens

In the present study, the loop-mediated isothermal amplification (LAMP) assay was developed to amplify the fragments of the O9 Salmonella-specific insertion element and evaluated in the laboratory for its potential use in a field situation, such as poultry farms. Among the bacteria tested, a positive reaction was observed only for 128 strains of 6 serovars of the O9 group Salmonella, such as Enteritidis (SE) and Pullorum. The detection limit of the LAMP assay was 10(3)CFU/ml, which was more sensitive than that of the polymerase chain reaction (PCR) assay with the same target gene (10(6)CFU/ml). The final results were obtained within 30 min for the LAMP assay, while the PCR assay needed a total of 120 min. When the LAMP assay was applied to the enrichment broth mixed with cecal dropping samples either spiked with SE in vitro or excreted by SE-inoculated hens, the results were comparable to those of the conventional plating method including 2 separate enrichments. In conclusion, the LAMP assay developed in the present study is an effective method for the specific detection of the O9 group Salmonella serovars, including SE.     

Etiologic study of upper respiratory infections of household dogs

Infectious tracheobronchitis (ITB), also known as the kennel cough, is a respiratory syndrome of dogs and usually appears to be contagious among dogs housed in groups. Etiologic agent of ITB is multiple and sometimes complex. In the present study, 68 household dogs showing clinical signs of respiratory infection were examined, and 20 dogs (29.4%) were found to be positive for either of following agents. Bordetella bronchiseptica (B.b.) was most frequently detected from nasal and oropharynx sites of 7 dogs (10.3%). Among the viruses examined, canine parainfluenza virus (CPIV) was detected with the highest frequency (7.4%). Other pathogens included in the order of frequency group 1 canine coronavirus (4.4%), canine adenovirus type 2 (2.9%), group 2 canine respiratory coronavirus (1.5%), and canine distemper virus (1.5%). Only 2 cases showed mixed infections. Neither influenza A virus nor canine bocavirus (minute virus of canines) was found in any dogs examined. These results indicate that both B.b. and CPIV are likely to be the principal etiologic agents of canine ITB in Japan, and they may be considered as the target for prophylaxis by vaccination.     

Intranasal hemangiosarcoma in a dog

Magnetic resonance (MR) was conducted for an 8-year-old, intact male Spitz with sneezing, serous discharge and epistaxis from the left nasal cavity. MR imaging showed a nasal cavity-occupied mass of iso-intensity on T1WI , high-intensity on T2WI and markedly enhanced on contrast-enhanced T1WI at parts of rostal to medial ocular angle in the left cavity. After Surgery and intraoperative radiation, the mass was diagnosed intranasal hemangiosarcoma by histopathology. Although the dog showed the finding, which suggested recurrence after the treatment ending, about 30 months later, it maintained good conditions without evidence of metastasis.     

Sex,age, or genetic differences related to play behaviors in Japanese Black calves

Play behavior in young animals has been used to evaluate the condition (health) of livestock. We investigated age, sex, and genetic differences of Japanese Black calves in relation to frequency of play behaviors (galloping, leaping, turning, bucking, head butting objects, and head shaking) and examined how these relationships might affect growth during the suckling stage. Locomotor play behaviors (galloping, leaping, turning, and bucking) and head butting objects gradually declined with the age for both sexes. The frequency of head butting was significantly higher in males than females. We found that significant interaction effects (age × MAOA polymorphism) in play behaviors (except head shaking) and the frequencies of locomotor play in calves without the wild‐type allele were significantly higher than those in younger calves (2 and 6 weeks of age). Weight gain was significantly correlated with the frequency of locomotor play in females, but not in males. This study suggests that play in Japanese Black calves gradually declines as they mature and that play may be controlled by variations in the MAOA gene. In addition, the frequency of locomotor play may be an indicator of health in female calves.     

Short-term transportation in a small vehicle affects the physiological state and subsequent water consumption in goats

We investigated the physiological effects on goats of transportation for one hour in a small truck. We also investigated food and water consumption after transportation. Eight adult goats (four castrated males and four ovariectomized females) were used. Plasma levels of cortisol, glucose and free fatty acids increased significantly within 15 min of the start of transportation, and these higher levels were maintained throughout transportation. These results indicate that transportation in a small truck activates the hypothalamus-pituitary-adrenal (HPA) axis and the sympathetic nervous system in goats. Despite activation of the sympathetic nervous system, heart rate (HR) in subjected animals during transportation did not differ from during the control animals, which were housed in their usual pen. However, after the transportation had ended and the goats started to eat fodder, HR increased significantly. There was no difference in the amount of food consumption between the control and transported goats. Water consumption in the first three hours after transportation was significantly lower in the transported goats than in the controls for the equivalent period. In the subsequent three hours, however, the transported goats consumed more water than the controls. Consequently, the total water consumption of the transported goats did not differ from that of the controls during the six hours after transportation. In conclusion, short-term transportation with a small vehicle activated the HPA axis and the sympathetic nervous system in goats. HR was not increased during transportation, but after the transportation had ended HR in transported goats was higher than that in control goats. After transportation, goats had consumed the same amount of food as control goats, but water consumption during three hours after the transportation was lower than that of control goats.     

Comparison of organic matter dynamics in soil between Japanese cedar (Cryptomeria japonica) forest and adjacent Japanese red pine (Pinus densiflora) forest established on flatland

In order to clarify the effects of tree species on organic matter dynamics in soil, we investigated the amount of forest floor material, leaf litter decomposition rate, soil chemical characteristics, soil respiration rate and cellulose decomposition rate in a Japanese cedar forest (cedar plot) and an adjacent Japanese red pine forest (pine plot) established on a flatland. The amount of forest floor material in the cedar plot was 34.5 Mg ha⁻¹ which was greater than that in the pine plot. Because the leaf litter decomposition rate was higher in the pine plot than in the cedar plot, it is likely that the difference in the amount of forest floor material between the plots is caused by the difference in the leaf litter decomposition rate. The C concentrations of soil in the cedar plot were 1.2–2.1 times higher than those in the pine plot. Soil pH(H₂O)s in the cedar plot were significantly higher than those in the pine plot. The soil respiration rates and the rates of mineralized C in the cedar plot byin vitro incubation were higher than those in the pine plot. From this result, it is assumed that soil organic matter in the cedar plot was decomposed relatively faster compared with the pine plot. Furthermore, microbial activities, which were reflected as cellulose decomposition rates in the cedar plot, were higher than those in the pine plot. A part of this paper was presented at the 109th Annual Meeting of the Japanese Forestry Society (1998).     

New sampler for dry deposition in forest canopy

A new type of sampler for dry deposition in a forest canopy was designed and tested to confirm its usefulness. The new sampler is made of Teflon chips packed in a nylon net, and tied on a rod like a branch in the canopy. The validity and characteristics of the new sampler were checked by comparing it with the conventional bucket sampler and the foliar extraction method using hinoki (Chamaecyparis obtusa) trees. The comparison between the deposited amounts on the dummy branch and those on the bucket indicates that the dummy could trap gaseous matter more effective than the bucket. From a comparison with the foliar extracting method, it is concluded that the trapping of particulate matter by the new dummy branch is comparable to that of natural needle leaves. However, its efficiency for gaseous matter is relatively small. This disadvantage should be improved by applying other materials to the dummy. Moreover, it is found that the uptake of gas especially SO₂ through the stomata is an important point in the discussion about dry deposition. To evaluate the amounts of dry deposition in the forest canopy more precisely, combination with the additional method should be useful. This study was supported in part by a grant-in-aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture, Japan.     

Relationships among the chalkiness,kernel size and endosperm cell morphology of rice kernels at different spikelet positions within a panicle

The occurrence of chalky kernels in rice is causally related to kernel size and endosperm morphology. This study aimed to investigate the occurrence of chalky kernels by analyzing kernel size, and the number and area of endosperm cells. Spikelets were sampled from upper and middle primary branches and lower secondary branches in a panicle, and divided into four categories: upper, middle, and lower perfect (PF) kernels and lower milky-white (MW) kernels. On the lower secondary branches, there was a higher percentage of chalky kernels, with smaller kernel lengths, widths and thicknesses, than the kernels on the upper and middle primary branches. MW kernels were smaller in size than PF kernels even on the same lower secondary branches. Regardless of grain appearance quality traits, the total areas of endosperm cross sections in lower kernels were significantly smaller than in upper kernels owing to the decreased cell area, and there was a significant negative correlation between the number of cells and average cell area. When the numbers and the areas of cells were analyzed using angular 30° intervals from the line connecting the center point and the dorsal vascular bundle, the MW kernels had significantly less cells than PF kernels near the ventral side at 120–180°. Thus, the decrease in the number of cells near the ventral side was a main causal factor in the decrease in MW kernel widths compared with PF kernel widths, and this suggested that cell division in MW kernels was inhibited at the early grain-filling stage.     

Starch Damage and Pasting Properties of Rice Flours Produced by Dry Jet Grinding

Milling method and particle size affect some properties of rice flour. To prepare ultra‐fine rice flour of <30 μm, hammer and dry jet grinding methods were examined and the effect of particle size on starch damage and pasting properties of the flour were elucidated. A jet mill could make finer flour (<10 μm mean size) with a narrower particle size distribution than a hammer mill could. Starch damage increased dramatically at a mean size of <10 μm. Particles of a similar size (<60 μm) had different levels of starch damage between mills. Not only the particle size, but also the milling method affected the level of damaged starch. Flour samples of ≥45 μm mean size had similar viscosity curves, but samples of <20 μm had different curves. Peak viscosity and final viscosity decreased sharply at <10 μm. Setback viscosity for particles of 3 μm from both brown rice and white rice were higher than the peak viscosity. Stability to heat and shearing stress were decreased for <20 μm flours as the breakdown viscosities decreased. Starch damage and pasting properties of flour ground from the nonwaxy japonica cultivar Koshihikari changed dramatically at a mean size of <10 μm.