S-genotype identification based on allele-specific PCR in Japanese pear

Gametophytic self-incompatibility in Japanese pear (Pyrus pyrifolia Nakai) is controlled by the single, multi-allelic S-locus. Information about the S-genotypes is important for breeding and the selection of pollen donors for fruit production. Rapid and reliable S-genotype identification system is necessary for efficient breeding of new cultivars in Japanese pear. We designed S allele-specific PCR primer pairs for ten previously reported S-RNase alleles (S¹–S⁹ and S^k) as simple and reliable method. Specific nucleotide sequences were chosen to design the primers to amplify fragments of only the corresponding S alleles. The developed primer pairs were evaluated by using homozygous S-genotypes (S¹/S¹–S⁹/S⁹ and S^4sm/S^4sm) and 14 major Japanese pear cultivars, and found that S allele-specific primer pairs can identify S-genotypes effectively. The S allele-specific primer pairs developed in this study will be useful for efficient S-genotyping and for marker-assisted selection in Japanese pear breeding programs.     

Ovulation of the preovulatory follicle originating from the first-wave dominant follicle leads to formation of an active corpus luteum

The objective of our study was to compare the characteristics of the corpus luteum (CL) formed after ovulation of the dominant follicle (DF) of the first follicular wave (W1) and those of the CL formed after ovulation of the DF of the second (induced) follicular wave (W2). Non-lactating Holstein cows were used for this study. In Experiment 1, cows were treated with PGF2α and GnRH on days 6 and 8 (day 0 = day of follicular wave emergence) for W1 (n = 6) and W2 (n = 6), respectively. Dominant follicles were aspirated on day 9 to quantify the amounts of mRNA (VEGF120, VEGF164, FGF-2, StAR, P450-scc and 3β-HSD) in granulosa cells (GC). In Experiment 2, the size and blood flow area of the CL formed after ovulation of the DF in W1 (W1CL; n = 6) and W2 (W2CL; n = 6) (the day of DF ovulation in W1 and W2 was day 10) were evaluated on days 12, 15, 18 and 21. The plasma P4 concentration was measured on days 10 to 21. The amounts of VEGF164, P450-scc and 3β-HSD mRNA were higher (P < 0.05) in the DF in W1, and those of VEGF120,FGF-2 and StAR mRNA tended to be higher (P < 0.1) in the DF in W1. The size of the CL was greater in the W1CL on days 15, 18 and 21. The blood flow area of the CL was greater in the W1CL on days 12 and 15. The plasma P4 concentrations were higher in the W1CL. These results indicate that the CL formed after ovulation of the DF in W1 was greater in terms of size, blood flow and plasma P4 concentration.     

Coincidental detection of genomes of porcine parvoviruses and porcine circovirus type 2 infecting pigs in Japan

The infection status of 15 viruses in 120 pigs aged about 6 months was investigated based on tonsil specimens collected from a slaughterhouse. Only 5 species of porcine parvoviruses and porcine circovirus type 2 (PCV2) were detected at high frequencies; 67% for porcine parvovirus (PPV) (PPV-Kr or -NADL2 as the new abbreviation), 58% for PPV2 (CnP-PARV4), 39% for PPV3 (P-PARV4), 33% for PPV4 (PPV4), 55% for PBo-likeV (PBoV7) and 80% for PCV2. A phylogenetic analysis of PPV3 suggested that Japanese PPV3s showed a slight variation, and possibly, there were farms harboring homogeneous or heterogeneous PPV3s. Statistical analyses indicated that the detection of PCV2 was significantly coincidental with each detection of PPV, PPV2 and PPV3, and PPV and PPV4 were also coincidentally detected. The concurrent infection with PCV2 and porcine parvoviruses in the subclinically infected pigs may resemble the infection status of pigs with the clinical manifestations of porcine circovirus associated disease which occurs in 3–5 months old pigs and is thought to be primarily caused by the PCV2 infection.     

Assessment of longitudinal tissue Doppler imaging of the left ventricular septum and free wall as an indicator of left ventricular systolic function in dogs

OBJECTIVE: To evaluate tissue Doppler imaging (TDI) of the left ventricular (LV) free wall (FW) and ventricular septum (VS) as an indicator of LV systolic function in dogs. ANIMALS: 7 healthy Beagles. PROCEDURES: Doses of dobutamine (5 and 10 microg/kg/min) and esmolol (50 and 100 microg/kg/min) were infused into the LV of each dog. With each dose, heart rate; myocardial performance index (MPI); transmitral inflow and ejection time (determined via pulsed-wave Doppler [PWD] echocardiography); and FW and VS velocities of the mitral valve annulus (determined via TDI during systole [S'], early diastole [E'], and late diastole [A']) were assessed. RESULTS: With each dose, dobutamine significantly increased heart rate and the first derivatives of LV pressure (+dP/dt and -dP/dt), whereas esmolol significantly decreased the +dP/dt and -dP/dt values, compared with baseline. Esmolol (100 microg/kg/min) significantly decreased the VS-TDI-derived S' velocity and FW-TDI-derived E' velocity; dobutamine significantly increased transmitral inflow and TDI velocities. Regression coefficient between VS-TDI-derived S' velocity and +dP/dt was higher than that between FW-TDI-derived S' velocity and +dP/dt. Compared with baseline, the PWD- and VS-TDI-derived MPI were significantly decreased by dobutamine and significantly increased by esmolol at each dose. Values of FW-TDI-derived MPI were higher than values derived via the other techniques. Correlation between +dP/dt and VS-TDI-derived MPI was greater than that between +dP/dt and FW-TDI- or PWD-derived MPI. CONCLUSIONS AND CLINICAL RELEVANCE: In healthy dogs, the VS-TDI-derived S' velocity and MPI appear to be reliable assessments for evaluating LV systolic function.     

Stock structure and resource management of hairtail <Emphasis Type="Italic">Trichiurus japonicus</Emphasis> based on seasonal broods around the Bungo Channel,Japan

Declines in landings of the hairtail Trichiurus japonicas indicate the need for more effective management of this species. Hairtail spawning peaks occur twice yearly in the Bungo Channel, in spring and autumn. Relationships between hairtail stock and brood seasonality were examined to determine if an association between either and a decline in landings existed. Stock assessments show that the biomass of both spring and autumn hairtail broods from within and around the Bungo Channel are decreasing, with a rapid reduction in spring-brood stock abundance after 2007 largely responsible for decreased landings. Yield and spawning per recruitment analyses indicate current fishing pressure to be higher than several reference points. We suggest that fishing pressure needs to be reduced by at least 20% of the current level for this fishery to remain sustainable, as the projected stock abundance and catch demonstrate that the current fishing pressure is unsustainable. Analysis of time-series data of recruits per spawning revealed spring-brood recruitment to have been strong in year classes 2003 and 2005. Of various options available for improved management of this fishery, we propose that fishing pressure should be reduced in the years following the appearance of strong year classes to increase future biomasses and landings.     

Genetic identification and morphology of naturally spawned eggs of the Japanese eel Anguilla japonica collected in the western North Pacific

Eggs of the Japanese eel Anguilla japonica collected in the western North Pacific were identified by onboard species-specific polymerase chain reaction (PCR) and DNA nucleotide sequencing after the cruise. Fish eggs of various species were collected by large plankton net tows at 12 stations along the southern part of the West Mariana Ridge on 19–25 May 2009. A total of 43 fish eggs were distinguished morphologically as possibly being of A. japonica. Thirty-one of those were analyzed by PCR, which included 15 eggs collected at 12°50–55′N, 141°15–20′E (in 5 tows) that showed positive results. The 16S ribosomal RNA (rRNA) gene sequences of eggs determined after the cruise indicated that 31 A. japonica eggs had been collected. The remaining eggs were of mesopelagic eel species (Serrivomeridae and Derichthyidae), or unidentified species. The morphological characteristics of the A. japonica eggs were consistent with those of artificially spawned eggs, except they had a slightly larger diameter. The egg diameter range did not overlap with those of mesopelagic eels of the Serrivomeridae, which often spawn in the same area as A. japonica. These results suggest that egg diameter and embryo shape can be used to morphologically identify naturally spawned A. japonica eggs.     

Vortex flow produced by schooling behavior of arabesque greenling <Emphasis Type="Italic">Pleurogrammus azonus</Emphasis>

Arabesque greenling Pleurogrammus azonus form schools of 30,000–60,000 individuals to feed on mesozooplankton near the sea surface. The fish need to be close to the surface where mesozooplankton production occurs, but the food is not sufficiently dense there for the fish to grow optimally while avoiding predation by sea birds. Arabesque greenling use a unique method to optimize their feeding conditions while avoiding staying on the sea surface. When the school swims upward, water is pushed downward. Upward swimming of a school of 30,000 individuals generates a downward stream of about 0.8–1.1 m/s, resulting in a convergent flow near the surface. This convergence concentrates the mesozooplankton on the sea surface and transports them into deeper layers with a strong vortex about 3.0 m in diameter and 10–20 m long. Thus, schooling of this fish induces vortex flows that provide a rich feeding environment.     

Estimating larval supply of Ezo abalone <Emphasis Type="Italic">Haliotis discus hannai</Emphasis> in a small bay using a coupled particle-tracking and hydrodynamic model: insights into the establishment of harvest refugia

Most juveniles of Haliotis discus hannai have been found to be descendants of wild individuals, although most adults were artificially produced (released) individuals as a result of restocking inside the refugium located near the head of Oshoro Bay, Hokkaido, Japan. To estimate the larval supply from released and wild individuals into the refugium and to compare the suitability of locations as larval sources, we simulated larval dispersal using a coupled hydrodynamic and particle-tracking model. The simulation results indicated that more larvae may be supplied from the wild adults inside the bay to the refugium than from the released adults. These results are consistent with the observed high abundance of wild juveniles in the refugium. Most larvae from the refugium were predicted to disperse out of the bay. We found that larval retention in the bay was at least one order of magnitude higher than that in the refugium. Thus, it may be more effective in terms of self-replenishment and reproduction if the refugium were to be expanded to the bay scale. There were only minor differences among the compared sites at the head of the bay in terms of their suitability as larval sources. Consequently, the establishment of new refugia in this area could be expected to provide an effectiveness equal to that of the current refugium.