Molecular characterization of the major outer membrane protein of Haemophilus somnus

The major outer membrane protein (MOMP) of Haemophilus somnus shows antigenic and molecular mass diversity that forms the basis of a preliminary grouping system for H. somnus strains. In this study, the gene encoding MOMP of H. somnus strain 8025 was cloned in three overlapping fragments by PCR techniques, and then sequenced. The gene consists of a 1164-bp open reading frame encoding a deduced 380-amino acid protein with a 19-amino acid signal sequence, giving a mature protein with a calculated molecular mass of 39,913 Da. Significant homology was found between MOMP and porin protein sequences of bacteria in Pasteurellaceae species. When expressed in Escherichia coli, the protein from the MOMP gene directed by the T7 promoter was identical in size (approximately 40 kDa) to native MOMP and reacted with MOMP-specific antibodies. Comparisons of the MOMP gene sequences from six unrelated strains of H. somnus to that of strain 8025 revealed that the genes of three MOMP type 1 strains were highly conserved with that of strain 8025 in length and sequence. However, two MOMP type 3c strains and one MOMP type 3a strain differed markedly from the MOMP of strain 8025 in their 3'-terminal halves. Their deduced MOMP amino acid sequences differed in sequence (3c, 80.5 and 82.7% identity; 3a, 62.4% identity) and in length (3c, 384 and 376; 3a, 316), indicating that the molecular differences are the basis of antigenicity and molecular mass differences of H. somnus MOMP. In the predicted MOMP secondary structure, the variable sequences primarily mapped to putative surface-exposed loops, and a variable and surface-exposed epitope of MOMP-specific antibody was identified in the seventh-largest loop. These findings are useful for understanding the structural and immunological characteristics of H. somnus.     

Identification of Facklamia sourekii from a lactating cow

A gram-positive, catalase-negative, facultatively anaerobic coccus was isolated from a lactating cow with hematuria and urodynia in Japan. The isolate was identified by 16S rRNA gene sequence analysis as Facklamia sourekii. The biochemical and culture characteristics of the isolate were well consistent with those of F. sourekii type strain. Since all F. sourekii strains reported so far were isolated from human clinical specimens, this is the first reported case of F. sourekii isolated from veterinary clinical specimen.     

Genetic diversity of Pinus densiflora pollen flowing over fragmented populations during a mating season

We attempted to evaluate the genetic diversity of long-distance transported pollen flowing over fragmented Pinus densiflora populations during a mating season. A P.?densiflora clonal seed orchard, which was located in a rural area where many fragmented populations exist, was selected for pollen capture. Immigrant pollen captured by three clones having different flowering times was regarded as the pollen flowing over fragmented populations during a mating season. The genetic diversity (H _e) values of the immigrant pollen captured by the three clones were high (H _e?>?0.894). The correlation of paternity (r _p) values of the seeds having immigrant parent generated from the three clones were calculated to be negative. From these parameters, the pollen cloud is considered to have maintained high genetic diversity during the mating season. The genetic composition of the pollen cloud showed slight variation. The pollen captured by different trees (i.e., clonal ramets of the three clones) was significantly different based on analysis of molecular variance. Especially, the pollen pools captured by trees planted in the western side of the orchard were significantly different from the gene pool of the surrounding populations. Factors affecting this differentiation could be that the donors of the pollen transported to the orchard vary with time, as well as nonuniform dispersal of the pollen. From these results, the pollen flowing over fragmented P.?densiflora populations is considered to have high genetic diversity, compensating to some extent for fragmentation.     

Spatial distribution pattern of attack of the oak borer,Platypus quercivorus (Murayama) (Coleoptera: Platypodidae), and scolytid ambrosia beetles (Coleoptera: Scolytidae) on fresh logs

The spatial distribution patterns of the attack on fresh logs ofPasania edulis was studied for the oak borer,Platypus quercivorus (Murayama), and two species of Scolytid ambrosia beetles,Xylosandrus crassiusculus (Motschulsky) andXyleborus attenuatus Blanford, in 1994 and 1995. On the logs where onlyP. quercivorus attacked, the entry holes were distributed uniformly when attack intensity was low. However, the distribution pattern became more aggregated with the increase in attack intensity. On logs where bothP. quercivorus and the two Scolytid species attacked, there was a negative association between the spatial distribution of the entry holes ofP. quercivorus and that of scolytids. Simultaneous attack of two scolytids also increased the degree of aggregation of the entry holes ofP. quercivorus. The entry holes of scolytids were distributed in groups irrespective of the attack intensity of scolytids andP. quercivorus. These results suggest an asymmetrical interspecific relationship betweenP. quercivorus and scolytids. Concentration of the entry holes ofP. quercivorus in a small area may cause a considerable decline in the reproductive success in the galleries constructed there.     

Detection of genetic variation among seed and seedlings ofChamaecyparis obtusa using allozyme markers

The effects of nursery practices on genetic composition and structure were studied inChamaecyparis obtusa by analyzing seeds and one-, two-, and three-year-old seedlings. Enzyme polymorphisms of the6Pgd-2, G6pd, Got, Gk, Shd-2, Pod, Dia-1, andPgm loci were used. OnlyDia-1 indicated significantly different allelic frequencies between seeds and three-year-old seedlings. Most of the genetic diversity of the parental clones in this seed orchard could be transferred to the offsprings. Genetic diversity on the basis of an average expected heterozygosity did not differ between seeds and seedlings. A slight excess of homozygotes in seeds and a significant excess of heterozygotes in seedlings of different ages to the expected panmictic proportions were found. Most of the differences in genotypic distributions among the seed and seedling stages, probably resulted from viability selection favoring outcrosses, and selective removal of inbred offsprings including self-fertilized ones prior to their transplantation in the nursery. A part of this paper was orally presented at the 107th Annual Meeting of the Japanese Forestry Society (1996).     

Estimation of the mode of inheritance of thermal tolerance in the guppy <Emphasis Type="Italic">Poecilia reticulata</Emphasis>

High water temperature influences the survival, growth, and maturation of fish. Genetically characterizing thermal tolerance is one of the most important subjects in fish culture. To identify the genetic characterization of thermal tolerance, this characteristic was compared among strains, and among parents and their offspring, in the guppy Poecilia reticulata. In the strain comparison, significant differences in survival rate were observed among the five strains examined, and between females and males. Females exhibited greater tolerance than males in four of five strains examined. In the comparison between parents and their offspring, stronger influence of female parent than of male parent was observed. Offspring obtained from surviving females exhibited greater tolerance than those from dead females. This tendency was typically observed in male offspring. The survival rate in male offspring obtained from dead female parents was lower than that of those from surviving females. The high-temperature tolerance of male parents did not influence this characteristic in offspring as strongly as that of female parents. These results suggest that the major gene or genes, which has a dominant resistant allele and a recessive sensitive allele, are probably passed on by sex-linked inheritance, located on the X chromosome.     

Effect of environmental factors and prey species on daytime and nighttime vertical distribution of juvenile walleye pollock in and around Funka Bay

We examined factors affecting the vertical distribution of juvenile walleye pollock during “the second critical period” (i.e. the transition period of prey size) in and around Funka Bay, Hokkaido, Japan. During the day, small juveniles (<30 mm) were distributed mainly in shallow, warm, and low-salinity water within the bay, whereas larger ones (≥30 mm) occurred in deeper, cold and saline water. During the night, the large juveniles migrated to shallower depths. Outside the bay, small- and large-sized juveniles were distributed in water masses of similar physical properties during the daytime, whereas during the nighttime, most of the juveniles moved to shallower depth, and some juveniles dived to deeper water mass. Dietary analysis revealed that large juveniles fed on large copepods (≥0.5 mm in body width) more than small juveniles did. The composition of zooplankton in our survey area showed that large-sized copepods often occurred in deeper saline water. From these results, there is a possibility that juvenile pollock inside and outside Funka Bay occupied different water masses preferable for each feeding advantage.