Effect of Harpin from Four Pathovars of Pseudomonas syringae on Pea Defense Responses

To investigate the role of the proteinaceous elicitor, harpin, on host and nonhost plants, we isolated the harpin-coding gene, hrpZ, from Pseudomonas syringae pvs. pisi, glycinea, tabaci and tomato. Effects of the recombinant harpin proteins on pea plants were analyzed and compared with the effects of the corresponding bacterial treatment. After inoculation of pea with pea pathogen P. syringae pv. pisi, the bacterial population increased and the accumulation of PAL-mRNA and pisatin was inhibited. The nonpathogenic pathovars, glycinea, tabaci and tomato induced both defense responses in pea. However, none of the harpins induced the hypersensitive reaction or accumulation of PAL-mRNA and pisatin in pea. Harpins from P. syringae pvs. glycinea, tomato and pisi did induce these defense responses in tobacco, however, suggesting that externally applied harpins either are not recognized or are nonfunctional in pea plants. Received 27 June 2000/ Accepted in revised form 21 February 2001     

Relationships Between the Appearances and Changes of Estrous Signs and the Estradiol-17β Peak,Luteinizing Hormone Surge and Ovulation During the Periovulatory Period in Lactating Dairy Cows Kept in Tie-stalls

Lactating Holstein-Friesian cows kept in tie-stall barn were used as subjects in this study. Rectal examination, ultrasonography and blood sampling were conducted every other day and then daily after the day on which diameter of the corpus luteum decreased. After the luteal diameter decreased for 2 consecutive days, rectal and ultrasound examinations, blood sampling, and observation of estrous signs were conducted at 6-h intervals. Most of the estrous signs became obvious with the increase in estradiol-17β (E₂) and became most remarkable 24 to 30 hours before ovulation, at which point the E₂ peak and luteinizing hormone (LH) surge were achieved, and then weakened which progression to ovulation. The correlation between the intensity of four estrous signs (hyperemia and swelling of the intravaginal part of the uterus, opening of the external uterine orifice and viscosity of the cervical mucus) and the plasma E₂ concentration was higher than that of three estrous signs (swelling of the vulva, contraction of the uterus, diameter of uterine horn) and the plasma E₂ concentration. The relaxation of the intravaginal part of the uterus showed a unique change compared with the other estrous signs, and it became most obvious 6, 12 and 18 h before ovulation; this obviously relaxed period was consistent with the generally accepted theoretical optimal time for artificial insemination (AI), i.e., 6 to 24 h after initiation of estrus. These results suggest that observation of estrous signs by vaginoscopic examination gave useful information for detection of the optimal timing of AI in the periovulatory period in lactating dairy cows kept in a tie-stall barn.     

Polymorphisms in the bovine hemoglobin‐beta gene provide evidence for gene‐flow between wild species of Bos (Bibos) and domestic cattle in Southeast Asia

The electrophoretic variation in bovine hemoglobin‐beta (HBB) is one of the most investigated genetic markers. The presence of a unique HBB variant, HBB^X, in Southeast Asian cattle has been reputed as a sign of gene‐flow from wild bovine species. In this study, we analyzed the DNA sequences of HBB genes in domestic and wild bovine species to verify this belief. Isoelectric focusing of HBB chain revealed that the HBB^X in domestic cattle had dimorphism and was separated into HBB^X1 and HBB^X2. The HBB^X1 had the same DNA sequence of the common HBB variant in gayal (Bos gaurus frontalis), while some of the HBB^X2 were identical with that of Cambodian banteng (Bos javanicus birmanicus). As a result, we confirmed that the bovine HBB variants can be a good indicator of introgression between wild and domestic cattle. The HBB^X1 was always predominant to HBB^X2 in the continental populations, suggesting that the gaur had contributed to the gene pool of domestic cattle in this region much more than the banteng. On the other hand, the mitochondrial DNA analysis could not detect gene‐flow from wild species. Autosomal markers that can trace the phylogeny between alleles are suitable for the assessment of bovine interspecific introgression.     

Concentrations of ketone body and antidiuretic hormone in cerebrospinal fluid in response to the intra‐ruminal administration of butyrate in suckling calves

The aim of the present study was to elucidate the mechanism by which ketone bodies increase antidiuretic hormone (ADH) secretion. Four male Holstein calves (5 weeks of age) were utilized. Four levels of butyrate (0 g, 11 g, 22 g and 44 g) were administrated intra‐ruminally in a 4 × 4 Latin square design and cerebrospinal fluid (CSF, six‐position lumbar puncture), blood plasma and urine were collected. The concentration of total plasma and CSF protein was 5.5–5.6 g/dL and 27.5–28.3 mg/dL, respectively. CSF concentrations of a specific ketone body, 3‐hydroxybutyric acid, were significantly higher in the 22 g and 44 g butyrate groups than in the control group. CSF concentrations of ADH in the 11 g and 44 g butyrate groups were significantly higher than in the control group. Plasma concentration of 3‐hydroxybutyric acid was increased by intraruminal administration of butyrate within 15 min in a dose‐dependent manner, and it was higher in the 22 g and 44 g butyrate group than in the control group from 15 min to 4 h. With the exception of the 11 g butyrate group, plasma concentrations of ADH also increased in response to butyrate treatment, and it was higher in the 44 g butyrate group than in the 22 g butyrate group from 15 min to 1.5 h. The duration of the elevated plasma concentrations of ADH was shorter than that of the plasma concentration of 3‐hydroxybutyric acid. The relationship between the plasma concentrations of ADH and 3‐hydroxybutyric acid was statistically significant but the correlation between the two concentrations was not high. Butyrate treatment elevated the plasma concentration of ADH and also resulted in reduced urine volume and increased urine osmolality. Haematocrit (Ht) values, and the osmolality of CSF and plasma were not different among the groups. Our results suggested that the increased ADH secretion observed in suckling calves fed dry feeds was caused by butyrate‐derived ketone body that crossed the blood‐brain barrier rapidly.     

Long-term stock assessment and growth changes of the Japanese sardine (Sardinops melanostictus) in the Sea of Japan and East China Sea from 1953 to 2006

We estimated the stock size of Japanese sardine ( Sardinops melanostictus ) in the Sea of Japan and East China Sea since 1953 using cohort analysis based on the changes of growth patterns. Growth of Japanese sardine, estimated by using annual rings on archived scales since 1961, showed that body lengths were extremely stunted in the 1980–1987 year-classes. The body length at age 3 from February to April in the 1980–1987 year-classes, a period when the stock size exceeded 4 million tons, was 180.0 ± 2.6 mm (mean ± SD), and in the other year-classes was 195.1 ± 7.6 mm. The body length at age 3 and wet weight of zooplankton in August in the offshore area of the Sea of Japan had a significantly positive correlation. We assumed three scenarios for maturation ratios, and estimated Ricker's spawner–recruitment relationships. We analyzed the correlations between logarithmic recruitment residuals (LNRR) and environmental factors in winter, represented by the North Pacific index (NPI), Aleutian low pressure index (ALPI), Pacific decadal oscillation (PDO), monsoon index (MOI), Arctic oscillation (AO) and Southern oscillation index (SOI). Significant correlations were observed between MOI and LNRR and between AO and LNRR. A combination of strong MOI and weak AO would increase the biomass of phytoplankton and zooplankton and subsequently increase the recruitment of Japanese sardine.     

Effects of calcium intake and parity on plasma minerals and bone turnover around parturition

Eight pregnant heifers (primiparous cows) and seven pregnant cows in their second, third and forth pregnancies (multiparous cows) were assigned to two groups and fed either a low calcium (Ca) diet (Ca, 0.46%) or a high Ca diet (Ca, 0.86%) ad libitum from 3 weeks before the expected calving date to 3 days after parturition. All cows were examined for a change in dry matter intake (DMI), plasma minerals and bone turnover around parturition. The dietary Ca level did not affect the DMI in both primiparous and multiparous cows. The DMI of primiparous cows was significantly lower than that of multiparous cows (P < 0.05) in both the low and high dietary Ca groups. The dietary Ca level did not affect the concentrations of plasma Ca, phosphorus, magnesium and parathyroid hormone throughout the experimental period. Plasma phosphorus in primiparous cows was significantly higher (P < 0.05) than that of multiparous cows around parturition. Plasma Ca and magnesium tended to be higher (P < 0.10) in primiparous cows. The dietary Ca level did not affect the plasma osteocalcin (OC) level measured as bone formation or the urinary deoxypyridinoline (DPD) excretion measured as bone resorption before parturition in both primiparous and multiparous cows. After parturition, the plasma OC level was lower than it was before parturition in multiparous cows fed a low Ca diet, and in primiparous cows. There were no significant differences in urinary DPD excretion between each group before and after parturition. Both the plasma OC level and urinary DPD excretion of primiparous cows were significantly higher than those of multiparous cows in both the low and high dietary Ca groups.     

Morphometric and histochemical study of involution in the rat uterus after parturition

Morphological changes to and collagen loss from the rat uterus during postpartum involution were investigated. The expression patterns of collagen type III and myeloperoxidase (MPO) were also determined. Morphological changes were studied on days 1, 3, 5, 10, 15, 20, 22 and 25 postpartum. As a control, diestrus rats’ uterine were used. Specimens from the uterine horn were embedded in paraffin, cut into 8 µm coronal sections, and stained with hematoxylin‐eosin. The thickness of the longitudinal and circular smooth muscle layers and of the endometrium were measured. The collagen content was determined using hydroxyproline analysis. Immunostaining was used to examine the expression of collagen type III on days 1, 3, 5, 10, 15 and 20 postpartum; and MPO on days 1, 3, 5, 10 and 22 postpartum. The thickness of the smooth muscle layers was found to decrease rapidly postpartum: the circular smooth muscle layer returned to that of a non‐pregnant, control uterus by day 5 postpartum and the longitudinal smooth muscle layer by day 15 postpartum. Eosinophilic cells were observed in the endometrial stroma adjacent to the myometrium on days 10, 15 and 20 postpartum, and were confirmed as collagenous cells. Immunostaining identified collagen type III positive cells in the vessel‐rich layer adjacent to the placental site on days 1, 3, 5 and 10 postpartum, and these cells were confirmed to be phagocytic. Postpartum reduction in the weight of the uterus was accompanied by decreases in both the collagen content and the thickness of the longitudinal and circular smooth muscle layers. Furthermore, the phagocytic cells were shown to express MPO during postpartum involution of the uterus.     

Central estrogen action sites involved in prepubertal restraint of pulsatile luteinizing hormone release in female rats

The present study aimed to determine estrogen feedback action sites to mediate prepubertal restraint of gonadotropin-releasing hormone (GnRH)/luteinizing hormone (LH) release in female rats. Wistar-Imamichi strain rats were ovariectomized (OVX) and received a local estradiol-17β (estradiol) or cholesterol microimplant in several brain areas, such as the medial preoptic area (mPOA), paraventricular nucleus, ventromedial nucleus and arcuate nucleus (ARC), at 20 or 35 days of age. Six days after receiving the estradiol microimplant, animals were bled to detect LH pulses at 26 or 41 days of age, representing the pre- or postpubertal period, respectively. Estradiol microimplants in the mPOA or ARC, but not in other brain regions, suppressed LH pulses in prepubertal OVX rats. Apparent LH pulses were found in the postpubertal period in all animals bearing estradiol or cholesterol implants. It is unlikely that pubertal changes in responsiveness to estrogen are due to a change in estrogen receptor (ER) expression, because the number of ERα-immunoreactive cells and mRNA levels of Esr1, Esr2 and Gpr30 in the mPOA and ARC were comparable between the pre- and postpubertal periods. In addition, kisspeptin or GnRH injection overrode estradiol-dependent prepubertal LH suppression, suggesting that estrogen inhibits the kisspeptin-GnRH cascade during the prepubertal period. Thus, estrogen-responsive neurons located in the mPOA and ARC may play key roles in estrogen-dependent prepubertal restraint of GnRH/LH secretion in female rats.     

Comparison of time budget of behaviors between penned and ranged young cattle focused on general and oral behaviors

To consider cattle welfare, time budget of behaviors of young cattle in pen and pasture conditions were compared. Behavioral observations of 103 steers in seven pens and 1136 steers in six ranges were performed during daylight over 3 days in each rearing condition. The pen condition had Japanese Black × Holstein cross, and the pasture conditions had assorted breeds of Angus, Murray Grey, Shorthorn, Hereford, Santa Gertrudis and their crosses aged 5–15 months. The ranges varied in vegetation from extensive native pasture to intensive improved pasture. The daytime proportion of behaviors and activity patterns were compared between rearing conditions. The proportion of walking was much lower in the pen condition (1.0 ± 0.2%) compared to all pasture conditions (at least 9.4 ± 4.0%; all P < 0.05), but grooming (5.9 ± 1.2%), investigating (2.5 ± 0.2%) and tongue playing (1.1 ± 0.2%) covered the loss. Fluctuation patterns of the proportions of eating (P < 0.001) and resting (P < 0.001) were different between pen and pasture conditions, whereas the pattern of the proportion of walking was not different. The proportion of eating in the pen condition (32.4 ± 0.3%) was not different to that in one on the farm (42.2 ± 12.6%), which had a thick improved pasture. However, the proportion of eating in the pen condition (32.4 ± 0.3%) was lower than that in two farms (58.8 ± 5.0% and 61.6 ± 16.5%) with low pasture availability (both P < 0.05). The proportions of oral behaviors such as self‐grooming (3.9 ± 1.0%), allogrooming (2.0 ± 0.2%), licking objects (1.3 ± 0.4%) and tongue‐playing (1.1 ± 0.2%) compensated for the lack of eating. These results indicate that we can learn a well‐balanced proportion of behaviors for cattle by making comparisons of time budget of behaviors and activity pattern between various rearing conditions. In addition, it is indicated that cattle under pen conditions under a restricted feeding period might compensate for a lower time spent feeding by performing other oral behaviors.