Evaluation of PCR as a means of identification of Pasteurella pneumotropica.

A polymerase chain reaction with new primers (new PCR) designed from Pasteurella pneumotropica 16S rDNA as an identification system for this organism was compared with the PCR reported by Wang et al. (Wang's PCR) by using 15 bacterial reference species and 70 clinical isolates with the conventional identification system. For the 15 reference strains, both PCRs were identical. For the 70 clinical isolates, the new PCR and Wang's PCR showed consistency with the conventional system in 62.9% (44/70) and 51.4% (36/70), respectively. Twenty-six isolates were inconsistent with the conventional system and the new PCR with respect to morphology and serology. These findings suggested that the new PCR was more sensitive than Wang's PCR, and the new PCR in combination with morphology and serology is useful for P. pneumotropica identification.     

Host specialization by reproductive interference between closely related herbivorous insects

Host specialization among closely related herbivores is common and is therefore a major theme in ecology. Despite its ecological importance, no general explanatory framework for host specialization is currently available. We introduce a new model of the evolution of host specialization in herbivorous insects. We use a two-dimensional lattice comprising 32 × 32 cells. Moreover, the model incorporates reproductive interference, defined as any negative outcomes resulting from interspecific mating, between two species of herbivores feeding on two different host plants as well as ordinary resource competition and differences in host suitability. Our simulation showed that reproductive interference, together with other factors such as host-related performance and resource competition, can actually drive the evolution of host specialization and thereby host partitioning between herbivorous insects during secondary contact. Host specialization arises particularly when levels of both reproductive interference and resource competition are intermediate. In contrast, host specialization seldom occurs if reproductive interference is absent. Thus, reproductive interference is as key to specialization as is spatial partitioning, which was not regarded as a realistic outcome when only resource competition was considered.     

Island biogeography as a test of reproductive interference

Theoretical studies have predicted that reproductive interference must exclude either of the interacting species, but no testing of this prediction has ever been reported for natural populations. This study surveyed the distribution patterns of herbaceous Veronica plants, including one native and three alien species, to test whether the invasion of the alien species exerting reproductive interference excluded the native species. Results showed that the native species was repeatedly excluded from islands where an alien species invaded, exerting reproductive interference, and that other alien species had no significant effect on the native population survival. This survey also demonstrated that the native species altered its habitat from the ground to stone walls on the mainland where the alien species had been predominant. In the mainland populations, the fruit morphology differed from that of the islands, and the morphology in the mainland population seemed suitable for seed dispersion by ants at a stone wall habitat. We also surveyed the genetic differentiation among populations, the results of which did not support the native species genetically differentiated between mainland and island populations before the alien species invasion. These results strongly suggest that the reproductive interference excluded the recipient species at the population level and facilitated the habitat change. Additionally, results indicated that a series of field surveys of islands close to the mainland can be a powerful tool to test the ecological importance of reproductive interference.     

Simple and on-site DNA purification for LAMP reaction applicable to non-adult tephritid fruit fly (Diptera: Tephiritidae)

Tephritid fruit flies are important pests of fruits and vegetables worldwide, but few studies have examined their ecology and life history in their native range because non-adult individuals of closely related species are morphologically indistinguishable. To identify non-adult individuals collected from host plants, flies examined in earlier studies were painstakingly reared to adulthood. Using Zeugodacus scutellatus Hendel as a model species, this study established a simple DNA extraction and purification method based on loop-mediated isothermal amplification (LAMP) reaction for on-site molecular identification of non-adult fruit flies. The non-adult individual body fluids were sampled using a sterilized toothpick and then diluted in TE buffer and heated for a short time. Alternatively, eggs were crushed in TE buffer using a sterilized toothpick and then heated. After heated buffer was added to the LAMP reaction mixture as template DNA and the mixture was heated for incubation of polymerase, the LAMP reaction detected the amplified DNA of Z. scutellatus. This purification method, characterized by heating after dilution with TE buffer, is simple and rapid. Our purification method, which obviates large centrifuges, thermal cyclers, and other apparatus, provides lower-cost and better on-site molecular identification of non-adult fruit flies than methods described in earlier reports. The LAMP-based molecular identification technique and our purification method are particularly useful for studying quarantined pest organisms that cannot be removed from their native areas.     

A serological survey on Bacillus piliformis infection in laboratory rabbits in Japan

A total of 544 rabbit sera obtained from 6 commercial breeding facilities and 9 research institutions during 1985-1990 were tested for Bacillus piliformis antibody by an enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescent antibody test (IFAT). The antibody was detected in 53 (14.2%) rabbits from 3 breeding facilities and 30 (17.4%) rabbits from 6 research institutions, indicating the prevalence of B. piliformis infection among laboratory rabbits in Japan. The overall agreement with ELISA for immune status was 96.9% (527/544) with IFAT. In tests of the ability of ELISA and IFAT to quantitate antibody, a correlation coefficient (r) of 0.86 (P less than 0.01) was obtained by plotting the measured average log of the ELISA titer against the corresponding log of the IFAT titer.     

Morphometric study of fat cell size in the fascia areolaris and fascia lamellaris of the inguinal region in men, women and pregnant women

The fat cells of the fascia areolaris and fascia lamellaris of men, women, and pregnant women (aged between 20 and 35a) were morphometrically studied. The cell volumes showed the following average values: 4.423 X 10(5) micron3 and 2.004 X 10(5) micron3 for the fasciae areolaris and lamellaris respectively, in men; 6.236 X 10(5) micron3 and 3.964 X 10(5) micron3 in women, and 10.114 X 10(5) micron3 and 4.635 X 10(5) micron3 in the pregnant women. The analysis of variance showed significant differences between both sexes, and fasciae areolaris and lamellaris. The differences between women and pregnant women as far as the cell volume is concerned, in both fasciae, were not significant. As to the fascia areolaris, not the lamellaris, the difference between the sexes was significant.     

Conflicting intersexual mate choices maintain interspecific sexual interactions

Reproductive interference, interspecific sexual interactions that affect reproductive success, is found in various taxa and has been considered as a fundamental source of reproductive character displacement (RCD). Once RCD has occurred, persistent interspecific sexual interactions between species pairs are expected to diminish. However, reproductive interference has been reported from some species pairs that sympatrically coexist. Thus, the question arises, can reproductive interference persist even after RCD? We modeled the evolutionary dynamics of signal traits and mate recognition that determine whether interspecific sexual interactions occur. Our models incorporate male decision making based on the recognition of signal traits, whereas most previous models incorporate only female decision making in mate selection. Our models predict the following: (1) even when male decision making is incorporated, males remain promiscuous; (2) nevertheless, the frequency of interspecific mating is maintained at a low level after trait divergence; (3) the rarity of interspecific mating is due to strict female mate recognition and the consequent refusal of interspecific courtship by females; and (4) the frequency of interspecific mating becomes higher as the cost to females of refusing interspecific courtship increases. These predictions are consistent with empirical observations that males of some species engage in infrequent heterospecific mating. Thus, our models predict that reproductive interference can persist even after RCD occurred.     

Detection of Sendai virus antibody in mouse and guinea pig sera by an enzyme-linked immunosorbent assay with protein A

An enzyme-linked immunosorbent assay using horseradish peroxidase (HRPO)-labeled protein A (P-ELISA) was established for detection of Sendai virus (SV) antibody in mouse and guinea pig sera. Sensitivity and specificity of P-ELISA were compared with those of ordinary ELISA using HRPO-labeled immunoglobulin G (IgG-ELISA) and the hemagglutination inhibition (HI) test. P-ELISA was 100 to 1,000 times more sensitive than the HI test for detection of the antibody in SV-naturally infected mice. P-ELISA and IgG-ELISA showed similar sensitivities for detection of the antibody in naturally infected mouse and guinea pig sera. A high specificity was demonstrated in P-ELISA with a cut-off optical density value of 0.2 (492 nm), while a non-specific reaction was observed when IgG-ELISA was used to both mouse and guinea pig sera at a low dilution (1:10-20). The antibody in rat sera was not detected by P-ELISA although it was realized by IgG-ELISA.     

Halo-shape technique for leadless pacemaker implantations: A case report

A 92-year-old woman underwent an implantation of a leadless pacemaker (Micra; Medtronic, Inc, Minneapolis, MN) for complete atrioventricular block after a transvenous lead extraction due to a pocket infection of a dual chamber pacemaker. Marked scoliosis and a humpback due to an advanced age made it impossible to direct the tip of the pacemaker delivery catheter towards the right ventricular septum or apex and shape the catheter into a gooseneck-shape. Thus, by attaining a halo-catheter shape of the delivery catheter, the catheter tip could be directed toward the infero-basal portion of the right ventricular septum. The pacemaker was successfully deployed at that site without any complications, and good device parameters were achieved. The halo-shape technique may be also an alternative method for delivering a leadless pacemaker in patients with an unsuccessful delivery of a leadless pacemaker to the right ventricular septum using the conventional gooseneck-shape technique.