Effect of rotifer enrichment with herbal extracts on growth and resistance of red sea bream,Pagrus major (Temminck & Schlegel) larvae against Vibrio anguillarum

The effects of the dietary medicinal herbs Massa medicata, Crataegi fructus (Cf), Artemisia capillaries, Cnidium officinale and their mixture (HM), on growth and resistance against Vibrio anguillarum were examined using larval red sea bream, Pagrus major. The methanol extracts of Cf and HM inhibited the proliferation of seven infectious bacterial strains including the genera Aeromonas, Edwasiella, Photobavterium, Pseudomonas and Vibrio. Rotifers enriched with emulsified bonito oil containing extracts of Cf and HM lowered the bacterial counts in their body on TCBS medium for the genus Vibrio in comparison with rotifers enriched by emulsified bonito oil alone. Although statistically not significant, larvae fed rotifer fortified with Cf from 3 to 20 days after hatching showed the best growth and final body weight followed by those given rotifer with HM and control enrichment. However, final body length of larvae fed rotifer with Cf was significantly larger than larvae fed on rotifers enriched with other herb extracts. After a challenge test with V. anguillarum, survival of larvae fed rotifer with Cf and HM was significantly higher than for the control treatment. These results revealed that Cf and HM in rotifer is useful to promote growth and resistance against V. anguillarum in red sea bream larvae, providing a new technology for mass production of disease‐resistant fry and fingerlings.     

Retrospective analysis of the efficacy of controlled internal drug release in follicular cysts in an embryo donor beef herd

We investigated the efficacy of a controlled internal drug release (CIDR) device in the reproductive management of an embryo donor beef herd. Superovulation of embryo donors was routinely induced by a combination of FSH and prostaglandin F(2)alpha analogue (PGF(2)alpha) at intervals of a few months, and after embryo recovery estrus of the donors was synchronized with PGF2(2)alpha. Between 1996 and 1998, approximately 20% of donors were diagnosed as having follicular cysts every year. Twenty-eight cases of follicular cysts recorded between 1997 and 1998 were treated with a CIDR device for 14 days to evaluate the efficacy of CIDR in resolving follicular cysts in donor herds. Initial recovery was defined as the occurrence of ovulation with estrous behavior and subsequent formation of a corpus luteum after removal of the CIDR. Initial recovery was recognized in all cases (n=28). Follicular cysts did not recur after repeated embryo recovery in 64% of the initially recovered donors, but in 36% of affected donors cysts recurred after the next embryo recovery. Subsequently, with a CIDR device instead of PGF(2)alpha, we synchronized estrus after embryo recovery in the same herd from 1999 to 2000, to investigate the ability of CIDR to prevent the initiation of follicular cysts. Of the donors used between 1999 and 2000, approximately 30% had a history of follicular cysts. Use of CIDR for estrous synchronization after embryo recovery lowered the incidence of follicular cysts to 3% in 1999 and 0% in 2000. Treatment with CIDR proved effective at resolving follicular cysts in the embryo donor beef herd and enabled re-use of donors affected with follicular cysts. CIDR is also likely to be efficacious in lowering the occurrence of follicular cysts in donor herds when it is used for estrous synchronization after embryo recovery.     

Availability of NMR microscopic observation of mouse embryo disorder: examination in malformations induced by maternal administration of retinoic acid

Nuclear magnetic resonance (NMR) microscopy is a magnetic resonance imaging method with enhanced spatial resolution due to the use of a high static magnetic field and high magnetic field gradients. It is considered to be a useful tool for non-invasive and continuous investigation of tissue and organs at the histological level. In this study, we applied NMR microscopy to assessment of morphology in mouse embryos using a developmental disorder model induced by retinoic acid administration. Pregnant mice were given 50 mg/kg all-trans retinoic acid at 8.5 dpc. Embryos were collected at several time points after treatment and examined by NMR microscopy after fixation. Two-dimensional and three-dimensional spin echo sequences were used. Tissue contrast on two-dimensional images changed according to length of repetition time and echo time, and also to developmental stage of embryos. Two-dimensional and three-dimensional images nondestructively demonstrated defects in development of the skeleton and soft tissue, e.g. hypoplasia of vertebrae in the lumbar and tail regions and dysplasia of the spinal cord, in embryos exposed to retinoic acid. These morphological abnormalities were confirmed by conventional assessment after imaging. Although further improvements are required, NMR microscopy will provide a new approach for multi-parameter assessment of embryonic development under physiological and pathological conditions.     

Decreased apoptotic polymorphonuclear leukocyte rate in dogs with pyometra

Polymorphonuclear neutrophil (PMN) apoptosis was examined in three dogs with pyometra by TUNEL assay in a 24-hr incubation period and compared with that in healthy control dogs (n=5). The incidence of apoptotic PMNs in dogs with pyometra was 26.4 +/- 5% and that in healthy dogs was 54.3 +/- 7%. The results indicated that apoptotic PMN rates in dogs with pyometra were significantly lower than those in control dogs (p<0.05), suggesting the prolongation of PMN survival.     

Ultrastructural study on the epithelial responses against attachment of indigenous bacteria to epithelial membranes in peyer's patches of rat small intestine

The ultrastructure of epithelial responses against the membrane adhesion of indigenous bacteria was investigated in the follicle-associated epithelium (FAE) of rat small intestine. The most frequent adherence of the various morphological types of bacteria to the epithelial membranes was found at the apex of the FAE. The attachment sites were deeply invaginated, and their bottoms were deformed into a sharp cone shape. Four layers with different electron densities were formed just beneath the apical membranes by microfilaments which surrounded the invaginations. The electron density of each layer was gradually decreased as being apart from the invaginations. The extremities of some bacteria in the invaginations were deformed into sharpened shapes. The cell walls of the extremities of the bacteria were occasionally dissolved in the invaginations, and their cytoplasms were slightly swollen with low electron densities. In some invaginations, the attached bacteria were eliminated to leave their fragments such as filamentous debris and a part of cell walls. Finally these remnants disappeared completely. When the bacterial colonies existed in the middle region of the FAE, the attachment of bacteria resulted in the engulfment of bacteria by M cells. The degenerated bacteria whose cytoplasmic matrices were separated into high electron dense materials and cleared materials were occasionally engulfed by ordinary microvillous columnar epithelial cells or goblet cells throughout the FAE. These findings suggest that the epithelial cells reject the attachment of live indigenous bacteria and that the M cells absorb indigenous bacteria in rat Peyer's patches.     

Plasma concentrations of inhibin A in cattle with follicular cysts: relationships with turnover of follicular waves and plasma levels of gonadotropins and steroid hormones

We investigated the profiles of circulating levels of inhibin A and total inhibin in beef cows with follicular cysts in relation to the patterns of follicular development and circulating gonadotropins and steroid hormones. Turnover of follicular waves was monitored in five cows every 2 days for 70 days from 10 days after detection of estrus without ovulation. The mean interwave intervals were 19.6 ± 1.0 days (n = 18 waves with cysts from the five cows). Circulating levels of inhibin A were approximately 170 pg/ml before emergence of follicular waves with cysts and increased (P < 0.05) concomitantly with follicle emergence. High concentrations of inhibin A (greater than 300 pg/ml) were noted for 7 days during the growth phase of cystic follicles, but inhibin A levels decreased gradually when development of the cysts reached a plateau. This profile of inhibin A was similar to those of total inhibin and estradiol, but was inversely related to the changes in plasma FSH concentrations. LH pulse frequency and mean concentrations of LH in cows with cysts were higher than those observed in the luteal phase of normal cyclic cows. These results indicate that the capacity to secrete inhibin, as well as estradiol, is maintained in cystic follicles, the growth of which is extended by LH secretion at levels greater than those seen in the normal luteal phase. Inhibin A plays an important role in the extension of interwave intervals by suppressing recruitment of a new cohort of follicles.     

Genetic variation and differentiation in tea (Camellia sinensis) germplasm revealed by RAPD and AFLP variation

Summary

Genetic variation and resulting variable groupings in tea (Camellia sinensis) and its wild Camelia relatives were assessed using Random Amplified Polymorphic DNA (RAPD) and Amplified Fragment Length Polymorphic (AFLP) markers. Numerous polymorphic bands were generated, of which 266 unambiguous ones were scored. The highest level of polymorphism as determined by the expected heterozygosity (H_av) was detected with AFLPs. Three major groups were recognized in the germplasm based on the parsimony method of cluster analysis. Two of the groups corresponded to varieties assamica and sinensis while the third group consisted of a heterogeneous mix of tea cultivars and related wild species. The Taiwan yamacha, unique tea cultivars grown predominantly in the highlands of Taiwan for the production of pan fired semi-fermented (Oolong and Pouchong) tea clustered in this group. Analysis of phenotypic diversity based on the generated RAPD and AFLP profiles revealed that population diversity (H), decreased in the order; Wild Camellia >India > China > Kenya > Sri-lanka > Vietnam > Japan > Taiwan. Analysis of molecular variance (AMOVA) revealed that most variation resided among individuals within populations (72%). Calculation of genetic distances and nested AMOVA further revealed a significant degree of structuring among the populations based on the country of origin.