Development of a method for detecting Coxiella burnetii in cheese samples

Coxiella burnetii is the causative agent of Q fever, and the main route of infection in humans is inhalation of contaminated aerosols. Although oral transmission by contaminated raw milk or dairy products is also a possible route of human infection, there have been few studies investigating the presence of C. burnetii in dairy products. We developed a new method of extracting DNA from cheese and detecting C. burnetii DNA in cheese samples with a nested PCR assay. The limit of detection was 6.0 × 10(2) C. burnetii particles per gram. We subsequently used this method to examine the presence of C. burnetii in cheese at commercial markets in Tokyo from June 2005 to December 2008. Twenty-eight of 147 cheese samples were found to be positive for C. burnetii DNA. However, when we assessed the viability of C. burnetii by inoculating mice with DNA-positive samples, all of the samples were found to be negative. Thus, the viability of C. burnetii appears to have been lost in these cheese samples.     

Assessment of radical scavenging capacity and lipid peroxidation inhibiting capacity of antioxidant

The role of radical scavenging antioxidants against oxidative stress has received much attention, and the antioxidant capacity has been assessed by various methods. Among them, a method that measures the effect of antioxidant on decay of the probe is one of the most widely used methods. The present study was performed to compare the two methods to assess the antioxidant capacity, one to follow the decay of the probe and the other to measure lipid peroxidation products in human plasma. It was shown that the method following probe decay was suitable for assessment of radical scavenging capacity of antioxidant, but not for the capacity to inhibit lipid peroxidation in plasma. This is true whether a hydrophilic or lipophilic probe is used. Such different results arise from the fact that the efficacy of inhibition of lipid peroxidation by antioxidants depends on the fate of antioxidant-derived radical and interaction between antioxidants as well as the capacity of free radical scavenging. Thus, the capacity of antioxidants for inhibition of lipid peroxidation should be assessed from the effect on the extent of oxidation, not from the effect on probe decay.     

ST segment depression and ventricular fibrillation in a dog after contrast agent administration

An 11-year-old Toy Poodle underwent a computed tomography examination with contrast (iohexol) enhancement under anesthesia. Heart rate and R-wave amplitude on electrocardiogram (ECG) increased 2.5 min after iohexol administration, and end-tidal carbon dioxide decreased to 12 mmHg. A progressive ST segment depression was observed on ECG. Subsequently, the ECG waveform changed to ventricular fibrillation. However, spontaneous circulation returned following cardiopulmonary resuscitation. Myocardial ischemia or anaphylactic shock was suspected in the dog, which explains the ST segment depression observed on ECG. When performing radiological examinations with a contrast agent, the ECG waveform changes, such as an increase in heart rate, R-wave amplitude, or ST segment depression, should be carefully monitored. This might enable early detection of cardiac dysfunction and the ensuing cardiac arrest in dogs.     

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).