Application of hot water emasculation to <Emphasis Type="Italic">Acacia auriculiformis</Emphasis> for controlled pollination

To determine the viability of hot water emasculation for Acacia auriculiformis, spikes composed of about 100 flowers were soaked in hot water at varying temperatures (45–60°C) and for varying lengths of time (5 s to 7 min) prior to anthesis. The rate of squashed polyads, each of which consisted of 16 pollen grains, and the germinating rate on media, decreased with temperature and time. A mating test suggested that the rates of adhering polyads per stigma and germinating polyads per stigma were lower in the self-pollination of soaked spikes than in that of normal spikes. On the other hand, the number of matured pods was not significantly different in the mating between the spikes receiving the soaking treatment (♀) and the spikes receiving no treatment (♂) compared with the mating between normal spikes. Although further research is needed to determine the optimal treatment conditions, these results suggest that soaking spikes into hot water, especially at 50°C for 45 s, 54°C for 15 s, and 56°C for 10 s, is an effective emasculation technique.     

PCR-based specific detection of <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> race 4 strains

Two primer sets were designed based on the sequence of polymorphic bands that were derived from repetitive sequence-based polymerase chain reaction (rep-PCR) fingerprinting and specifically detected in Ralstonia solanacearum race 4 strains (ginger, mioga, and curcuma isolates). One primer set (AKIF-AKIR) amplified a single band (165bp) from genomic DNA obtained from all mioga and curcuma and some ginger isolates; another set (21F-21R) amplified one band (125bp) from the other ginger isolates. These primer sets did not amplify the bands from genomic DNA of other R. solanacearum strains or of other related bacteria. PCR detection limit for the pathogen was 2 × 10²cfu.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession numbers AB118756 and AB118757     

Spinal cord effects from lumbar myelographic injection technique in the dog

To characterize spinal cord effects of needle placement using lumbar puncture myelography technique, lumbar puncture was performed in 5 dogs and computed tomography images of the spinal column were acquired in the transverse plane at the level of the puncture site after contrast injection and both before and after needle removal. The spinal cords were punctured during needle placement and parenchymal contrast enhancement was present in 4 of 5 dogs. Although no dogs exhibited overt neurological abnormalities following computed tomographic imaging, hemorrhage, gliosis and axonal degeneration were confirmed microscopically in all subjects. These results suggest that spinal cord morbidity is induced when lumbar myelography is performed using currently accepted technique.     

Relationship between growth hormone (GH) pulses in the peripheral circulation and GH-releasing hormone and somatostatin profiles in the cerebrospinal fluid of goats

Growth hormone (GH) is secreted in a pulsatile manner, but the underlying mechanisms of GH pulse generation remain to be resolved. In the present study, we investigated the relationship between GH pulses in the peripheral circulation and GH-releasing hormone (GHRH) and somatostatin (SRIF) profiles in the cerebrospinal fluid (CSF) of male goats. The effects of an intracerebroventricular (icv) injection of neuropeptide Y (NPY), galanin and ghrelin were also analyzed. Blood and CSF samples were collected every 15 min for 8 hr from the jugular vein and third ventricle, respectively. GH pulsatility in the goat was found to consist of distinct large pulses of 5 hr periodicity and small pulses of 1 hr periodicity. GHRH and SRIF in the CSF fluctuated in a pulsatile manner with 1 hr periodicity, and most of the descending phase of SRIF pulses were associated with the initiation of GH pulses. Icv injections of NPY, galanin and ghrelin stimulated GHRH release without affecting SRIF release. In addition, NPY suppressed, and galanin and ghrelin induced large GH pulses, although ghrelin was much more effective than galanin. These results suggest that an hourly fall in SRIF is involved in generating intrinsic circhoral rhythm of GH pulsatility. The mechanisms underlying the generation of large GH pulses of 5 hr periodicity remain unknown, while direct action of NPY and/or ghrelin on the pituitary might be involved.     

OPTIMIZING TECHNICAL CONDITIONS FOR MAGNETIC RESONANCE IMAGING OF THE RAT BRAIN AND ABDOMEN IN A LOW MAGNETIC FIELD

The present study was designed to establish the appropriate technical conditions for magnetic resonance imaging (MRI) of the head and abdomen in rats using a low magnetic field strength (0.2 T) MRI unit equipped with three radio frequency (RF) coils: a custom-made solenoid coil, a temporomandibular joint surface coil and a knee quadrature (QD) RF coil. Male adult Sprague-Dawley rats were used. T1 and T2 relaxation times of both anatomic regions were measured from T1 and T2 maps. An appropriate parameter was then used to make final T1 and T2 weighted images. It was found that the most suitable coil for the head was the custom-made soleniod coil, and that for the abdomen, the knee QD coil. The T1 and T2 relaxation times were 314 to 316 msec and 72 to 74 msec for the head, and 220 to 252 msec and 42 to 51 msec for the abdomen. The optimum parameters for the head were TR/TE = 400/38 msec in T1 weighted images and TR/TE = 1,800/110 msec in T2 weighted images, and for the abdomen, TR/TE = 300/25 msec in T1 weighted images and TR/TE = 1,500/110 msec in T2 weighted images. These results demonstrate that a low magnetic field strength MRI unit has potential for MRI study of the brain and abdomen in rats.     

Methane emission from stored dairy manure slurry and slurry after digestion by methane digester

To examine the effect of storage temperature on the emission of methane (CH₄) and carbon dioxide (CO₂) and on recovery of nutrients from raw dairy manure slurry (RS) and slurry digested in a methane digester (DS), both slurries were stored in closed 100 L steel tanks under psychrophilic conditions (5, 10, 15 and 20°C) for a 150 day period. As the storage temperature increased, total methane emission increased in both types of slurry. The amount of methane emitted per unit of volatile solids of the RS and DS was 0.19 L/g and 0.10 L/g, respectively. The respective carbon dioxide emissions were 0.20 L/g for RS and 0.12 L/g for DS at 20°C of storage temperature. At temperatures greater than 15°C, the methane concentration in the emitted gas remained more than 40% of the total gas. During the experimental period, in excess of 90% of the total Kjeldahl nitrogen in these slurries was recovered (91.4–93.7% for RS and 93.7–98.4% for DS) after storage, and ammonium nitrogen was recovered in excess of 100% (100.1–143.2% for RS and 106.7–143.2% for DS storage tanks) because of the mineralization of organic nitrogen in the influent. These results indicate that manure slurry characteristics and storage temperature have significant impacts on methane emission. It can be concluded that on typical farms located in northern Japan, methane emission from manure storage tanks during late fall, winter and early spring may be negligible, because of manure temperatures less than 10°C. During late spring, summer and early fall, methane emissions can be substantially reduced by using underground storage to maintain lower manure temperatures.     

A comparison between injection speed and iodine delivery rate in contrast-enhanced computed tomography (CT) for normal beagles

The purpose of this study was to compare between the injection speed and iodine delivery rate in order to establish a concept for reproducible contrast timing in contrast-enhanced computed tomography (CT) for small animals. Clinically healthy beagle dogs were administered a nonionic iodinate contrast medium at a dose of 800 mgI/kg; they were divided into 3 groups (n=5, crossover method): in one group, the injection speed was fixed at 1.0 ml/sec, and in the second and third groups, the iodine delivery rate was fixed (the injection durations were 30 and 60 sec, respectively). The variation in scatter of the time to aortic and hepatic peak enhancement in the fixed iodine delivery groups was lower than that in the fixed injection speed group. These results suggest that in contrast-enhanced CT for small animals, the contrast medium should be injected at a fixed iodine delivery rate in order to provide reproducible contrast timing.     

Brain abscess in a Japanese black calf: utility of computed tomography (CT)

Computed tomography (CT) was used for diagnosis of brain abscess in a 6-month-old, Japanese black calf presented with neurological dysfunction, compulsive circling and vision disturbance. CT images showed asymmetric lateral ventricles, and presence of intra-cranial multiple low absorption lesions surrounded by capsule suggestive of abscess in the right cerebral hemisphere. Postmortem examination revealed marked swelling of right cerebral hemisphere and olfactory bulb. Multilocular large abscess containing creamy pus was found to occupy most area of periventricular and lateral ventricle. Fusobacterium necrophrum was isolated from the abscess contents as the causative agent. These results demonstrate that CT is useful tool for tentative diagnosis of bovine brain abscess.     

Assessing diastolic function with Doppler echocardiography using a novel index: ratio of the transmitral early diastolic velocity to pulmonary diastolic velocity

We developed a novel index to assess left ventricular (LV) relaxation as the ratio of transmitral early diastolic velocity to pulmonary diastolic velocity (E/D ratio). Mixed breed dogs (n=7) were anesthetized and their respiration was controlled. A 3.5-Fr micromanometer-tipped catheter was placed into the left ventricle. Dobutamine (5.0 or 10 microg/kg/min) or esmolol (100 or 500 microg/kg/min) was administered via the cephalic vein. The transmitral flow (TMF) and pulmonary venous flow (PVF) were recorded using transthoracic echocardiography from the apical long-axis view. The heart rate, systolic LV pressure, +dP/dt, and -dP/dt were significantly elevated by dobutamine, but significantly reduced by esmolol. Dobutamine significantly decreased tau, whereas esmolol significantly increased tau. The TMF-derived E and PVF-derived D wave velocities increased significantly with dobutamine, but decreased significantly with esmolol. A significant correlation was detected between the E and D wave velocities (r=0.92). Consequently, the E/D ratio was decreased significantly with dobutamine, and increased significantly with esmolol. Furthermore, the E/D ratio was significantly correlated with -dP/dt (r= -0.64) and tau (r=0.84). Our results suggest that the E/D ratio reflects LV relaxation, and may potentially provide further information on LV relaxation.     

Central administration of neuropeptide B, but not prolactin-releasing peptide, stimulates cortisol secretion in sheep

Two neuropeptides, neuropeptide B (NPB) and prolactin-releasing peptide (PrRP), have been suggested to play important roles in control of the hypothalamic-pituitary-adrenal (HPA) axis in rodents. The aim of the present study was to clarify the central actions of NPB or PrRP in sheep. Ovariectomized ewes were surgically implanted with a cannula directed to the lateral ventricle. They received intracerebroventricular (icv) administration of 400 mul of artificial cerebrospinal fluid, NPB (0.05, 0.5 or 5 nmol), PrRP (0.5, 5 or 50 nmol) or corticotropin-releasing hormone (CRH, 0.5 or 5 nmol) through the cannula, and blood samples were taken 30 and 0 min prior to and 15, 30, 60 and 90 min after the injection. Cortisol concentrations in plasma were determined by enzyme immunoassay. Administration of 0.5 nmol NPB resulted in a significant increase in the cortisol concentration compared with the vehicle control, whereas the cortisol concentration after lower or higher doses of NPB did not differ from the control value. Thus, an icv injection of NPB produced a bell-shaped dose-response of cortisol concentration. Administration of PrRP had no significant effect on the cortisol concentrations at any dose examined. Icv injection of CRH dose-dependently increased plasma cortisol concentrations. These results demonstrate that central NPB stimulates cortisol secretion, suggesting that this neuropeptide plays some roles in control of the HPA axis in sheep. On the other hand, unlike its role in rodents, PrRP is unlikely to be involved in control of the HPA axis in this species.