Molecular cloning of equine muscle-type phosphofructokinase cDNA

The complete coding region sequence of equine muscle-type phosphofructokinase (ePFKM) was obtained from skeletal muscle of a thoroughbred horse. The deduced amino acid sequence of ePFKM showed 97%, 96%, 96%, 96% and 95% identity to canine, human, mouse, rabbit and rat PFKM, respectively. The amino and carboxyl terminal halves of ePFKM presented a structure of tandem repeat, as other mammalian PFKMs. As the amino acid residues constituting various ligand-binding sites were also conserved, it is thought that ePFKM has enzymatic activity similar to PFKM in other mammals.     

Molecular cloning and expression of bottlenose dolphin (Tursiops truncatus) interleukin-8

The bottlenose dolphin interleukin (IL)-8 cDNA was molecularly cloned. The dolphin IL-8 has an open reading frame of 303-bp encoding 101 amino acids. The homology of the amino acid sequence with that of other species was: sheep, 89.1%; cattle, 88.1%; pig, 85.1%; dog, 85.1%; horse, 79.2%; human, 74.5%; and macaque, 72.3%. The amino acid sequence suggested that dolphin IL-8 was a CXC chemokine. The recombinant dolphin IL-8 protein was recognized with anti-ovine IL-8 monoclonal antibody.     

Microsatellite polymorphism in Japanese mongrel dogs

The genetic variability of 182 unrelated mongrel dogs living in various areas of Japan (from Hokkaido to Okinawa) was studied by collecting their blood. Ten microsatellite loci were chosen from different autosomal chromosomes. After combining a few rare adjoining alleles to allelic classes, it was confirmed that the Hardy-Weinberg equilibrium was attained in each locus. The polymorphic information contents (PICs) of the loci, Ren37A11, Ren48E01, AHTk253, ZuBeCa30, Ren277K09, Ren42N13, AHT130, PEZ03, PEZ12, and AHT121, were 0.58, 0.63, 0.67, 0.67, 0.68, 0.71, 0.79, 0.80, 0.80, and 0.80, and the power of discriminations (PDs) were 0.80, 0.85, 0.87, 0.88, 0.88, 0.89, 0.94, 0.94, 0.94, and 0.94, respectively. The combined mean exclusion chance (MEC) was 0.9995, indicating that these microsatellite loci are useful for kinship testing of Japanese dogs.     

Exposure of C57BL/6J male mice to an electric field improves copulation rates with superovulated females

It is well-known that there are considerable strain differences in the relative copulation rates between male and superovulated female mice. In particular, the C57BL/6J strain of mice has a lower rate of successful copulation. We examined the effect of exposure to an electric field on sexual behavior in C57BL/6J male mice. When C57BL/6J males were exposed to a 50 Hz, 45 kV/m electric field for 30 min per day for 11 days and placed in a cage with a superovulated female of the same strain, the successful copulation rates of males was significantly improved compared with unexposed males (P<0.05). These results suggest that the exposure of C57BL/6J male mice to an electric field improves their sub-fertility activity in mating with superovulated females.     

Effective pollination period in durian (Durio zibethinus Murr.) and the factors regulating it

The effective pollination period (EPP) of durian was determined by both delayed and bud pollination, during which reproductive factors affecting the EPP, e.g., stigma receptivity, pollen tube growth in the style, and ovule longevity were studied histologically. This study was conducted in three distinct locations in Thailand, namely, the Chantaburi Horticultural Research Center and two private orchards in Chantaburi and Trat provinces. Results from artificial pollination revealed that at anthesis, the durian flower is receptive and has a high fruit set ratio. A mean fruit set of 50% was obtained at anthesis in the private orchard in Chantaburi province. However, the EPP of durian was found to be very short, lasting for only one night; the fruit set from pollination on the morning after anthesis ranged from 0% to 3.4%. No fruit set occurred following pollination 24 or more hours after anthesis. When compared with the flowers of other fruit species, the durian flower has a unique feature in that it blooms overnight; the following morning, there is abscission of all parts of the flower, except the gynoecium. Thus, EPP appears to be synchronized with flower longevity. On the other hand, the durian flower was receptive several hours before anthesis. The results of chemical tests, including the hydrogen peroxide test and Perex-Test^®, for the evaluation of stigma receptivity appeared to be in agreement with the EPP. However, fluorescent microscopy revealed that pollen could germinate even in the stigmas pollinated 48 h after anthesis, but the number of pollen tubes at the top of the style rapidly decreased from 34.6 (at anthesis) to 0.5 (48 h after anthesis). A correlation test demonstrated a higher correlation coefficient between the fruit set and the number of pollen than that between the fruit set and the result of Perex-Test^®. This indicated that pollen tube penetration or elongation in the style was inhibited, probably due to the deterioration of nutritional support from the pistil to the pollen tubes; this can be a limiting factor of the EPP in durian.     

Identification of sialyl oligosaccharides including an oligosaccharide nucleotide in colostrum of an addax (Addax nasomaculatus) (Subfamily Antelopinae)

Mammalian milk/colostrum usually contains milk oligosaccharides along with the predominant lactose. Although milk oligosaccharides of a variety of Bovidae species including cow, sheep and goat have been characterized, those of the addax, an Antelopinae species of the Bovidae, have not as yet been clarified. In this study, several sialyl oligosaccharides were purified from a sample of addax colostrum and characterized as follows: Neu5Ac(α2‐8)Neu5Ac(α2‐3)Gal(β1‐4)Glc, Neu5Gc(α2‐8)Neu5Gc(α2‐3)Gal(β1‐4)Glc, Neu5Ac(α2‐3)Gal(β1‐4)Glc, Neu5Ac(α2‐6)Gal(β1‐4)GlcNAc, Neu5Gc(α2‐3)Gal(β1‐4)Glc, Neu5Gc(α2‐6)Gal(β1‐4)Glc, Neu5Gc(α2‐6)Gal(β1‐4)GlcNAc. In addition, an oligosaccharide nucleotide Neu5Gc(α2‐6)Gal(β1‐4)GlcNAcα1‐UDP was characterized. Molecular species of a variety of sialyl oligosaccharides found in milk and colostrum of these Bovidae were compared.     

The effects of protein sources supplemented with urea-treated potato pulp (PP) silage and feeding levels of the PP silage-based concentrate on feed intake, digestibility and ruminal fermentation in beef steers

Six Japanese Black (Wagyu) steers (average initial weight 467 ± 45 kg) fitted with a ruminal cannula were used in a split‐plot design experiment comprising a 3 × 3 Latin square design (whole‐plot) and a randomized block design (subplot). The whole‐plot treatments were three different feeding levels of urea‐treated potato pulp (PP) silage‐based concentrate: 1.00%, 1.75% and 2.50% of body weight (BW) (on a dry matter (DM) basis). The subplot treatments consisted of the concentrate formulated to contain either soybean meal (SBM) as a rapidly rumen‐degraded protein source or corn gluten meal (CGM) as a slowly degraded protein source. Dry matter intake tended to be lower (P = 0.071) for CGM (8.9 kg/day) than for SBM (9.4 kg/day). Protein sources had no significant effect on digestibility and in situ degradation. Ruminal ammonia nitrogen (NH₃‐N) was lower (P = 0.033) for CGM (7.5 mg/dL) than for SBM (9.5 mg/dL). Protein sources did not affect ruminal pH and the total volatile fatty acids (VFA) concentrations. The molar proportions of ruminal acetate and valerate were higher (P = 0.032) for CGM than for SBM. The maximum daily intake of the PP silage‐based concentrate expressed as a percentage of BW was approximately 1.4% of BW. Dry matter intake was higher (P = 0.046) for steers fed at 1.0% of BW of the PP silage‐based concentrate than for steers fed at 1.75% or 2.5% of BW of the concentrate. The feeding levels of the PP silage‐based concentrate had no effect on DM and nutrients digestibility, except for crude protein (CP) digestibility. CP digestibility tended to be lower (P = 0.071) for steers fed at 1.75% of BW of the PP silage‐based concentrate than for steers fed at 1.0% or 2.5% of BW of the concentrate. The feeding levels of the PP silage‐based concentrate also did not affect the in situ degradation parameter of hay and PP silage. The feeding levels of the PP silage‐based concentrate did not affect ruminal pH, NH₃‐N and total VFA concentrations. The molar proportion of acetate was highest for steers fed at 1.0% of BW of the concentrate. In conclusion, in the urea‐treated PP silage‐based concentrate, CGM seems to be more effective than SBM for stabilizing the ruminal NH₃‐N concentration and to be advantageous for fiber digestion in the rumen. The feeding levels of the PP silage‐based concentrate did not change the amount of VFA production in the rumen and the DM digestibility.     

Evaluation of basal media for micropropagation of four highbush blueberry cultivars

Murashige and Skoog medium (MS), woody plant medium (WPM), and a mixture of equal parts of MS and WPM (MW) were compared for in vitro shoot proliferation and rooting of four highbush blueberry (Vaccinium spp.) cultivars. During the multiplication stage, the shoots on WPM showed worse growth than the other shoots. The MW produced the best shoot growth. The shoots on MS grew well but tended toward hyperhydricity in the multiplication stage, especially in the case of ‘Bluecrop’ shoots. Rooting was worst in the shoots multiplied on WPM, while the best rooting percentage of ‘Bluecrop’ shoots was obtained on MS, and that of ‘O’Neal’ on MW. The rooted shoots developed a good root system and were easily acclimatized after potting.     

Establishment of a laboratory colony of taiga tick Ixodes persulcatus for tick-borne pathogen transmission studies

Ixodes persulcatus Schulze (I. persulcatus) is distributed in Russia and Far East Asia including Japan, and has been implicated as the vector of several human pathogens. In particular, I. persulcatus acts as the only tick vector for human lyme borreliosis in Japan. In order to elucidate the mechanism of transmission of I. persulcatus-borne pathogens, we developed a laboratory colony of I. persulcatus. Ticks were fed on Syrian hamster and engorged ticks that had dropped off the animals were collected and maintained to allow them to molt. Tick rearing was performed in incubator at 20 degrees C with 95% relative humidity and 12-hour light/dark photo-period regimen. We found out that adult females fed for 8 +/- 2 days and had a pre-oviposition period lasting for 7 +/- 2 days. The minimum egg incubation period was 1 month with the hatched larvae feeding for 3 +/- 1 days and molting to nymphs 3-4 months thereafter. Meanwhile, the nymphs fed for 4 +/- 1 days and molted to adult 2-3 months thereafter. For future analysis of gene expression profiles in I. persulcatus, we cloned and sequenced the actin gene (a housekeeping gene), and found that it is 92.7% to 98.6% homologous to the published sequences of related ixodid ticks. This laboratory colony of I. persulcatus will facilitate investigations on the role of tick-derived molecules on the transmission of I. persulcatus-borne pathogens and will be important for identification of potential anti-tick vaccine and acaricide target molecules.     

Cushing's disease complicated with thrombosis in a dog

Thrombosis is a potential complication of hyperadrenocorticism (HAC) in dogs. An 8-year-old male Beagle diagnosed with pituitary-dependent HAC had complicated with thrombosis in the caudal vena cava and abdominal aorta, which was treated by hypophysectomy and antithrombotic therapy. After hypophysectomy, hypercortisolemia disappeared and the general condition was also significantly improved. Ultrasonography after hypophysectomy revealed that the thrombus remained in the abdominal aorta, but the thrombus in the caudal vena cava had disappeared. However 692 days after the hypophysectomy, the dog had an acute onset of dyspnea and died. Postmortem examination revealed the presence of thrombi in the abdominal aorta and the pulmonary artery. Observations from this case show that HAC dogs must be attention to thrombosis.