Cloning of estrogen receptor-β cDNA of common Japanese conger Conger myriaster and expression of the gene during oocyte development

ABSTRACT:   Estrogen receptor (ER) cDNA was cloned from the liver of common Japanese conger Conger myriaster . Common Japanese conger ER cDNA contains 2543 nucleotides including an open reading frame that encodes 596 amino acids, with a molecular weight of 66.1 kDa. The deduced amino-acid sequence of the common Japanese conger ER shares 85% identity with that of eel Anguilla japonica ER-β, and 39–46% and 53–59% identity with the deduced sequences of ER-α and ER-β of other species, respectively. In a phylogenetic analysis, the common Japanese conger ER was found to cluster with the ER-β of other species. The common Japanese conger ER-β also has six domains (A–F), which are conserved in the nuclear receptor superfamily. The amino-acid sequences in the C and E domains were highly conserved between common Japanese conger ER-β and other ER. However, the other domains showed low homology. In the female common Japanese conger, ER-β mRNA was highly expressed in the liver and pituitary gland. The expression levels of the ER-β gene increased from the oil droplet stage to the primary yolk globule stage and were maintained until the migratory nucleus stage.     

Effect of bovine lactoferrin feeding on lipopolysaccharide-induced metabolic and hormonal disturbances in preruminant calves

One of the biological functions of bovine lactoferrin (LF) is modulation of the host defense system, including cytokine production and immune response. The aim of the present study was to investigate the effect of oral administration of LF in calves on lipopolysaccharide (LPS)‐induced metabolic and hormonal changes in inflammatory response. Thirty Holstein calves at 4 day of age were given one of three oral doses of LF (0, 1, 3 g/day) for 10 days (?10 day to ?1 day). They were injected i.v. with LPS (50 ng/kg bodyweight) the day (day 0) after the end of LF treatment. Plasma samples were obtained on ?10, 0 day (immediately before LPS injection), and at 2, 6, 12, 24, 48, 72, and 96 h after LPS injection. Plasma tumor necrosis factor‐α concentrations at 2 h after LPS treatment were lower (P < 0.05) in LF 1 g/day‐fed claves compared with LF 0 g/day (control) calves. On day 0 there were no significant group differences in plasma LF concentration. Plasma concentration of haptoglobin in control calves was elevated by LPS injection. In LF groups, plasma haptoglobin concentrations slightly increased after LPS injection, but those levels at 6–24 h were lower (P < 0.05) than in the control group. The LF treatment inhibited (P < 0.05) the reduction of plasma ferrin concentration in calves following LPS challenge. The concentration of plasma aspartate aminotransferase in calves treated with LF was lower (P < 0.05) than in control calves at 24–96 h after LPS treatment. The concentration of plasma insulin‐like growth factor‐1 (IGF‐1) in all groups was decreased by LPS treatment, while in the LF groups the IGF‐1 level was higher (P < 0.05) than in the control group. Plasma adrenocorticotropic hormone and insulin concentrations in LF groups were lower (P < 0.05) than in control calves at 2 h after LPS injection. These data suggest that LF has a substantial anti‐inflammatory effect on the modulation of the host defense system in preruminant calves.     

Citrus hybrid seedlings reduce initial time to flower when grafted onto shiikuwasha rootstock

The long juvenile period of citrus seedlings before flowering is one of the major obstacles in citrus breeding. Under the citrus breeding program of National Institute of Fruit Tree Science in Japan, 299 citrus hybrid seedlings were grafted onto shiikuwasha (Citrus depressa HAYATA) and cultivated. Approximately one-third of the plants flowered two and a half years after grafting. The average length of the main stems of flowering plants was 308 cm, while that of non-flowering plants was 264 cm. Half of the plants having main stem length ≥300 cm formed flowers, while most of the plants having main stem length <210 cm formed no flower. These results indicate that plant vigor influences the flowering and that grafting effectively accelerated flowering. The proportion of flowering plants varied among cross combinations, ranging from 71.4% to 8.0% among the 10 cross combinations used.     

A canine case with cystic meningioma showing miraculous reduction of the cystic lesion

A 12-year-old spayed female Labrador retriever was presented with forebrain signs. Brain MRI revealed a huge cystic lesion with the thickened falx in the frontal region. The brain parenchyma surrounding the lesion showed significant signs of a mass effect and also increased intracranial pressure. However, the dog suddenly became lucid after about two weeks, and an MRI scan one month after the initial study revealed a dramatically shrunken cystic lesion. The dog survived for over a year until it was euthanized for other reasons, and the brain lesion was diagnosed as a cystic meningioma histologically. To the authors’ knowledge, this is the first report that described the reduction of the cystic lesion of a cystic meningioma in dogs.     

Oral antibiotics enhance antibody responses to keyhole limpet hemocyanin in orally but not muscularly immunized chickens

Recent studies have emphasized the crucial role of gut microbiota in triggering and modulating immune response. We aimed to determine whether the modification of gut microbiota by oral co‐administration of two antibiotics, ampicillin and neomycin, would lead to changes in the antibody response to antigens in chickens. Neonatal chickens were given or not given ampicillin and neomycin (0.25 and 0.5 g/L, respectively) in drinking water. At 2 weeks of age, the chicks were muscularly or orally immunized with antigenic keyhole limpet hemocyanin (KLH), and then serum anti‐KLH antibody levels were examined by ELISA. In orally immunized chicks, oral antibiotics treatment enhanced antibody responses (IgM, IgA, IgY) by 2–3‐fold compared with the antibiotics‐free control, while the antibiotics did not enhance antibody responses in the muscularly immunized chicks. Concomitant with their enhancement of antibody responses, the oral antibiotics also lowered the Lactobacillus species in feces. Low doses of antibiotics (10‐fold and 100‐fold lower than the initial trial), which failed to change the fecal Lactobacillus population, did not modify any antibody responses when chicks were orally immunized with KLH. In conclusion, oral antibiotics treatment enhanced the antibody response to orally exposed antigens in chickens. This enhancement of antibody response was associated with a modification of the fecal Lactobacillus content, suggesting a possible link between gut microbiota and antibody response in chickens.     

Domestication and cereal feeding developed domestic pig‐type intestinal microbiota in animals of suidae

Intestinal microbiota are characterized by host‐specific microorganisms, which have been selected through host‐microbe interactions under phylogenetic evolution and transition of feeding behavior by the host. Although many studies have focused on disease‐related intestinal microbiota, the origin and evolution of host‐specific intestinal microbiota have not been well elucidated. Pig is the ideal mammal model to reveal the origin and evolution of host‐specific intestinal microbiota because their direct wild ancestor and close phylogenetic neighbors are available for comparison. The pig has been recognized as a Lactobacillus‐type animal. We analyzed the intestinal microbiota of various animals in Suidae: domestic pigs, wild boars and Red river hogs to survey the origin and evolution of Lactobacillus‐dominated intestinal microbiota by metagenomic approach and following quantitative PCR confirmation. The metagenomic datasets were separated in two clusters; the wild animal cluster being characterized by a high abundance of Bifidobacterium, whereas the domesticated (or captured) animal cluster by Lactobacillus. In addition, Enterobacteriaceae were harbored as the major family only in domestic Sus scrofa. We conclude that domestication may have induced a larger Enterobacteriaceae population in pigs, and the introduction of modern feeding system further caused the development of Lactobacillus‐dominated intestinal microbiota, with genetic and geographical factors possibly having a minor impact.     

Activity of cyazofamid against Sclerospora graminicola, a downy mildew disease of pearl millet

The efficacy of cyazofamid was tested against pearl millet downy mildew disease caused by Sclerospora graminicola Schroet. Significant inhibition of sporangial sporulation, zoospore release and motility was observed at 0.3 mg mL(-1), and this concentration also provided good fungicidal activity under in vitro conditions. Under glasshouse conditions, none of the concentrations tested, either 0.01-2 mg mL(-1) as seed treatment or 1-10 mg mL(-1) by foliar application, was found to be phytotoxic. The effect of cyazofamid was tested by seed treatment alone, seed treatment followed by foliar application and foliar application alone. Seed treatment with cyazofamid offered only 19.7% disease control, but seed treatment followed by a single foliar application to diseased plants provided good control over disease, seed treatment with two foliar applications was significantly superior and foliar application alone showed a high level of activity, with 10 mg mL(-1) giving 97.9% disease control. Lack of systemic activity of cyazofamid was evident, root treatment giving disease levels on a par with the untreated control. The fungicide exhibited strong curative activity, but only moderate translaminar activity, with only marginal (34.8%) disease control after treatment of the adaxial leaf surface at 10 mg mL(-1). Loss of cyazofamid activity over time was very low, indicating stable residual and rainfastness activity. These results indicate that cyazofamid has a high potential to be an effective fungicide for the control of downy mildew disease of pearl millet.     

Role of C. elegans TAT-1 protein in maintaining plasma membrane phosphatidylserine asymmetry

The asymmetrical distribution of phospholipids on the plasma membrane is critical for maintaining cell integrity and physiology and for regulating intracellular signaling and important cellular events such as clearance of apoptotic cells. How phospholipid asymmetry is established and maintained is not fully understood. We report that the Caenorhabditis elegans P-type adenosine triphosphatase homolog, TAT-1, is critical for maintaining cell surface asymmetry of phosphatidylserine (PS). In animals deficient in tat-1, PS is abnormally exposed on the cell surface, and normally living cells are randomly lost through a mechanism dependent on PSR-1, a PS-recognizing phagocyte receptor, and CED-1, which contributes to recognition and engulfment of apoptotic cells. Thus, tat-1 appears to function in preventing appearance of PS in the outer leaflet of plasma membrane, and ectopic exposure of PS on the cell surface may result in removal of living cells by neighboring phagocytes.     

Characteristics of nutrient load in a stream flowing through a livestock farm during spring snowmelt

We investigated the stream water quality during the snowmelt period in a livestock farm, located in Shizunai, southern Hokkaido, Japan. The water quality was very different between the early stage of the snowmelt period (March 15–21) and the later stage (March 22–April 5) in 2001. The load of nutrients (NH₄ ⁺-N, T-P, K⁺) was large along with the increase of the flow rate and nutrient concentrations during the early stage of the snowmelt period. The molar ratios of Si / T-N and Si / T-P of the stream water, as an index of eutrophication, were also under the threshold values (2.7, 64.3, respectively) during the early stage of the snowmelt period. In addition, the relationship between the Si and nutrient concentrations (NH₄ ⁺-N, T-P, K⁺) showed a significantly negative correlation (R ² = 0.65), which indicated that the amount of nutrient load might be derived from surface runoff. Consequently both the quantity and quality of the stream water during snowmelt may exert an adverse effect on coastal waters, leading to eutrophication. Furthermore, since the main effluent source appeared to be surface runoff at the early stage of the snowmelt period, attention should be paid to land management before and during the snow covering period.