Differentiation of uterine natural killer cells in pregnant SCID (scid/scid) mice

To determine whether functional T- and B-cells can affect differentiation and/or proliferation of uterine natural killer (uNK) cells, their numbers in SCID mice (genotype, C.B.-17/Icr-scid/scid) were compared with those of control mice (genotype, C.B.-17/Icr-+/+) on days 8, 12 and 16 of pregnancy. Using biotinylated-Dolichos biflorus agglutinin (DBA) lectin staining, uNK cells can be readily classified into 4 subtypes, I to IV, from immature to mature types. The number of uNK cells was significantly lower in the decidua basalis of SCID mice than in that of control mice on day 8 of pregnancy. Particularly, the number of uNK cells of immature subtype II was significantly lower in SCID mice than in the control mice. By day 12, however, the uNK cell number in the SCID mice reached the same level as that of the control mice. It is likely that uNK cell differentiation in SCID mice was delayed during the early placentation period due to a lack of functional T and B cells.     

Effects of repeated atropine injection on heart rate variability in Thoroughbred horses.

To investigate the effects of repeated atropine injection on heart rate (HR) variability in resting Thoroughbred horses, two microg/ kg of atropine as parasympathetic nervous blockade was injected intravenously every 6 min to a total of 8 microg/kg after intravenous administration of 0.2 mg/kg of propranolol as sympathetic nervous blockade. We recorded electrocardiograms and obtained the HR, then evaluated variation in HR from the power spectrum in terms of low frequency (LF, 0.01-0.07 Hz) power and high frequency (HF, 0.07-0.6 Hz) power. Administration of atropine decreased parasympathetic nervous activity in a dose-dependent manner, affecting first the LF power, then the HF power and finally HR. These responses may provide valuable information for evaluating autonomic nervous activity in Thoroughbred horses.     

Polymyxin B enhances acrosomal exocytosis triggered by calcium and the calcium ionophore A23187 in ejaculated boar spermatozoa

Polymyxin B (PMB) is beneficial for boar semen storage since it neutralizes the endotoxin of bacteria. However, the direct effect of PMB on boar spermatozoa has been unknown. This study aimed to examine the effect of PMB on acrosomal exocytosis, an essential process for successful fertilization in boar spermatozoa. Ejaculated spermatozoa stored with BTS extender at 17°C were washed and incubated with 0–100 μM PMB for 20 min and then examined for % total motililty, vigor grade and viability. None of the parameters was significantly different between 0 and 50 μM PMB with a gradual decline at higher concentrations. Thus the effect on acrosomal exocytosis was investigated at 0–50 μM of PMB. Spermatozoa were preincubated with PMB for 10 min, incubated for stimulation of acrosomal exocytosis with Ca²⁺ and the calcium ionophore A23187 and then fixed with glutaraldehyde at 5, 10 and 15 min. Preincubation with PMB at 0.01–50 μM and 0.05–50 μM resulted in significant enhancement of acrosomal exocytosis at 10 min and 15 min of incubation, respectively. Preincubation with PMB followed by incubation without A23187 did not affect acrosomal exocytosis. These results suggest that PMB exerts effects on the acrosomal exocytosis triggered by Ca²⁺ and A23187 in boar spermatozoa.     

Long‐term impact of puerperal metritis on the profiles of peripheral blood leukocytes in peripartum dairy cows

To determine the effects of puerperal metritis on the immune response, changes in the differential peripheral blood leukocyte counts were analyzed during the peripartum period in cows with or without metritis. Multiparous Holstein cows were examined for uterine health disorders and classified into two groups: healthy (n = 11) or metritis (n = 5) cows. The lymphocyte and monocyte counts and the proportion of CD8⁺ lymphocytes were higher in cows with metritis compared to healthy cows. Moreover, the effects of puerperal metritis on the lymphocyte counts and CD4⁺/CD8⁺ ratio persisted weeks after the uterine inflammation had self‐resolved. Taken together, the findings of the present study indicate the possible long‐term alterations of systemic immune responses in cows with puerperal uterine inflammation. © 2015 Japanese Society of Animal Science     

Sex Pheromone of the Almond Moth and the Indian Meal Moth: cis-9, trans-12-Tetradecadienyl Acetate

Female moths of different species but belonging to the same subfamily produce an identical compound as their sex pheromone. The sex pheromone of the almond moth, Cadra cautella (Walker), and the Indian meal moth, Plodia interpunctella (Hübner), has been isolated and identified as cis-9, trans-12-tetradecadienyl acetate.     

Genetic diversity of African wild rice (Oryza longistaminata Chev. et Roehr) at the edge of its distribution

Ethiopia is at the edge of the distribution for African wild rice, Oryza longistaminata. Here, chloroplast (cp) and simple sequence repeat (SSR) markers were applied to wild rice accessions in Ethiopia to evaluate how they differ from control O. longistaminata, O. barthii and O. glaberrima accessions which originated from African countries. Based on the cp genomes of African wild rice species, maternally inherited cpINDEL markers were developed. The cp indels helped to elucidate 20 plastid types. African cultivated rice shared a particular plastid type with one of annual O. barthii. Parts of northern wild rice in Ethiopia shared Type 6 with control O. longistaminata. The north group shared another type with parts of the south group. The 16 SSR markers amplified a total of 155 alleles in 215 rice accessions, with mean allelic richness of 9.688 per locus, observed heterozygosity of 0.241, expected heterozygosity of 0.724, polymorphic information content of 0.700, and a significant genetic differentiation of 0.215. Both cpINDEL and nuclear markers analyses suggested that wild rice in Ethiopia belongs to O. longistaminata. However, they carry both a unique plastid type and different population structure from control O. longistaminata collected from other areas in Africa. We concluded that the edge of its distribution maintains unique variation. These populations are regarded as valuable genetic resources for future rice breeding.

     

Development of artificial model of caval syndrome in canine heartworm disease

In order to develop an artificial model of caval syndrome (dirofilarial hemoglobinuria), heartworm-like silicone tubes were inserted into the tricuspid valve orifice and right atrium of dogs. Fifteen to 25 tubes with some knots were inserted through the posterior vena cava in 6 dogs (knot-tube group), 7 to 12 tubes without knot (small-number group) through the jugular vein in another 5 dogs, or 25 to 35 tubes (large-number group) in yet another 5 dogs. The tubes remained in the right atrium, and a part of them protruded into the tricuspid valve orifice. The number of tubes at the tricuspid valve orifice was the greatest in the large-number group. After tube insertion, the signs of so-called "caval syndrome", such as systolic cardiac murmur, jugular pulse, anemia, and so on, were observed in almost all cases of the 3 groups, the signs were severest in the large-number group. Urine hemoglobin was detected in almost all cases of the knot-tube and large-number groups, and in 1 case in the small-number group. Ascites was observed in 1 case of the knot-tube group at 6 weeks, in 1 case of the small-number group at 7 days and in 3 cases of the large-number group at 7 days after insertion.     

Multiplication of isolate R-5 of <Emphasis Type="Italic">Streptomyces galbus</Emphasis> on rhododendron leaves and its production of cell wall-degrading enzymes

 An endophytic actinomycete, isolate R-5 of Streptomyces galbus Frommer, that has promising potential as a biocontrol agent was originally isolated from field-grown rhododendron. In this study, the mode of entry of R-5 into leaves of tissue-cultured seedlings of rhododendron was investigated in connection with its production of cell wall-degrading enzymes. Light and scanning electron microscopy (SEM) revealed that R-5 grew on leaf surfaces and entered leaf tissues via stomata and that the internal mycelia grew out of stomata after colonization in host tissues. Micromanipulation at the SEM level demonstrated a prominent depression in the host surface at the interfaces with the mycelia, suggesting that such a depression could be caused by degradation of cell wall components by hydrolytic enzymes secreted from R-5 mycelia. In subsequent plate assays, R-5 produced cellulase, pectinase, xylanase, and nonspecific esterase when cultured in liquid medium. Moreover, R-5 multiplied in mineral medium containing cellulose, pectin, or xylan as a single carbon source. Thus, R-5 mycelia could degrade host cell walls at contact sites and probably utilize the degradation products as carbon sources. Received: May 16, 2002 / Accepted: July 9, 2002     

Toward the restoration of a sustainable population of a threatened aquatic plant, Nymphoides peltata: Integrated genetic/demographic studies and practices

We provide a brief review of practices and relevant studies for restoration of Nymphoides peltata (yellow floating heart, Asaza), a distylous floating-leaf clonal plant, in Lake Kasumigaura, Japan to develop a model of integrated genetic and demographic conservation for threatened plants. Several theoretically expected issues, such as sudden extinction of small populations with low genetic diversity, limited seed production in small-sized local populations, and significantly high heterozygosity in adults that have survived environmental change, were ascertained through integrated studies on demography with discrimination of genets and genetics using highly polymorphic genetic markers. Investigation of genetic properties of the remnant soil seed bank suggested that the seed bank could potentially restore genetic diversity, although the fitness reduction of seed banks caused by inbreeding could affect the success of restoration. As a result of restoration efforts, increases in the number of local populations and genets in the Lake Kasumigaura metapopulation have led to population recovery.     

SEM observations on larval shell morphology of Manila clam Ruditapes philippinarum and their utility

Manila clam Ruditapes philippinarum is common and abundant on Japanese tidal flats, forming a commercially important clam fishery. However, annual catches of Manila clam have decreased drastically since 1975?C1985. To study larval recruitment processes of Manila clam, we carried out scanning electron microscopy (SEM) observations on larval and juvenile shells of clams reared at 20 and 24?°C. There was no significant difference in final shell length of trochophores between 20 and 24?°C. However, the larval duration was much longer and the shell length of settled size of pediveligers was much larger for clams reared at 20?°C than those reared at 24?°C. These findings suggest that larval duration and growth, as well as settlement size, may vary markedly depending on temperature (and probably on season). The larval shell morphology of Manila clam can provide essential information about larval recruitment processes.