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Social specificity: interaction with own species is necessary to foster species-specific maternal preference in ducklings.

Previous studies have shown that if domestic mallard ducklings are allowed social interaction with broodmates after their initial exposure to a stuffed mallard (Anas platyrhynchos) hen, they later show a visual preference for the familiar mallard over an unfamiliar redhead (Aythya americana) hen. Birds kept in social isolation do not make this difficult discrimination. In the present study, we examined whether this social enhancement of imprintability to a natural maternal model is merely the result of social stimulation in its most general sense, or if its effects are specific to social interaction for 30 min (training). This experience resulted in visual preference for the familiar mallard hen over the unfamiliar redhead hen in simultaneous choices tests at 48 hr and 72 hr after hatching only if the birds were allowed social experience with a group of same-age undomesticated mallard hatchings. No visual preference for the familiar mallard hen was found if the ducklings were permitted social experience with a group of same-age chick (Gallus domesticus) hatchlings or muscovy (Cairina moschata) ducklings. Thus, it appears that for ducklings to show a visually imprinted preference for a natural maternal model, they must be allowed social interaction with broodmates of their own species. Social rearing with other precocial bird hatchlings, even other ducklings, is without apparent effect. These results provide further evidence of the importance of nonobvious experimental factors in the development of species-typical behavior. They also suggest that in the evolution of the species-specific learning underlying early species identification, natural selection has operated on the entire (organism-environment) developmental manifold and not merely on genetic, organic, or organismic aspects of that system.

Animals↗

Stage-specific and species-specific differences in the production of the mRNA and protein for the filarial nematode secreted product, ES-62.

Previous studies have shown that the secreted phosphorylcholine-containing glycoprotein of filarial nematodes, ES-62, is only present in the post-infective life-cycle stages, but that the mRNA is transcribed throughout the worm's life-cycle. The aim of this current study was to investigate whether the presence or absence of protein expression simply reflects differences in mRNA abundance. To this end, we investigated the relative abundance of ES-62 using TaqMan real time RT-PCR, in different life-cycle stages of 2 model filarial nematode parasites, Acanthocheilonema viteae and Brugia pahangi. For B. pahangi, microfilariae, infective larvae and adult worms were each found to have approximately similar levels of ES-62 mRNA. However, the corresponding stages of A. viteae differed greatly from each other with a pattern of increased mRNA production with maturation. As a rule A. viteae had higher levels of ES-62 mRNA than B. pahangi, and this was particularly noticeable in the adult stage where the difference was approximately 3500-fold higher. However, this significant difference in mRNA abundance was not reflected in the quantity of ES-62 protein secreted by the adult worms of each species, as A. viteae only secreted approximately 3 times as much ES-62 as B. pahangi. Thus, overall, the results obtained from this study indicate that ES-62 protein production does not solely reflect mRNA levels, and also suggest that the 2 nematodes may employ different mechanisms for regulating protein production.

Animals↗

Characterization of species-specifically amplified SINEs in three salmonid species--chum salmon, pink salmon, and kokanee: the local environment of the genome may be important for the generation of a dominant source gene at a newly retroposed locus.

Short interspersed repetitive elements (SINEs), known as the HpaI family, are present in the genomes of all salmonid species (Kido et al., Proc. Natl. Acad. Sci. USA 1991, 88: 2326-2330). Recently, we showed that the retropositional efficiency of the SINE family in the lineage of chum salmon is extraordinarily high in comparison with that in other salmonid lineages. (Takasaki et al., Proc. Natl. Acad. Sci. USA 1994, 91: 10153-10157). To investigate the reason for this high efficiency, we searched for members of the HpaI SINE family that have been amplified species-specifically in pink salmon. Since the efficiency of the species-specific amplification in pink salmon is not high and since other members of the same subfamily of SINEs were also amplified species-specifically in pink salmon, the actual sequence of this subfamily might not be the cause of the high retropositional efficiency of SINEs in chum salmon. Rather, it appears that a highly dominant source gene for the subfamily may have been newly created by retroposition, and some aspect of the local environment around the site of retroposition may have been responsible for the creation of this dominant source gene in chum salmon. Furthermore, a total of 11 sequences of HpaI SINEs that have been amplified species-specifically in three salmon lineages was compiled and characterized. Judging from the distribution of members of the same-sequence subfamily of SINEs in different lineages and from the distribution of the different-sequence subfamilies in the same lineage, we have concluded that multiple dispersed loci are responsible for the amplification of SINEs. We also discuss the additional possibility of horizontal transmission of SINEs between species. The availability of the sets of primers used for the detection of the species-specific amplifications of the SINEs provides a convenient and reliable method for identification of these salmonid species.

Animals↗

Tissue-specificity overrides species-specificity in cytoplasmic cytochrome c oxidase polypeptides.

With a high-resolving dodecyl sulfate electrophoretic system rat liver cytochrome c oxidase was separated into 13 different polypeptides. An antiserum against rat liver holocytochrome c oxidase immunoreacted with all 13 polypeptides, as demonstrated by immunofluorescence after transfer of the separated Coomassie blue-stained bands on nitrocellulose and coupling with FITC-protein A ("western blot"). Polypeptide-specific antisera reacted only with their corresponding polypeptides indicating that the various protein bands are represented by individual polypeptides. From total proteins of rat liver, kidney, heart, spleen and skeletal muscle mitochondria, only the cytochrome c oxidase polypeptides showed immunofluorescence with an antiserum against the rat liver holoenzyme. In contrast to the polypeptide from liver, polypeptide VIa from heart and skeletal muscle showed little or no reactivity, indicating a tissue-specificity of this polypeptide. Mitochondrial proteins from pig, bovine and blackbird heart were incubated with an antiserum against the rat liver holoenzyme. Immunoreaction was found with most cytochrome c oxidase polypeptides but not with polypeptide VIa. This result demonstrates less immunological relationship between tissue-specific polypeptides (VIa, VIIa and VIII) of the same species than between tissue-unspecific polypeptides of different species.

Animals↗

Identification of beta-subunit of bacterial RNA-polymerase--a non-species-specific bacterial protein--as target of antibodies in primary biliary cirrhosis.

Several observations suggest that bacteria induce autoimmunity in primary biliary cirrhosis (PBC). Since no PBC-specific bacterial species could be identified, it can be speculated that the triggers are non-species-specific bacterial proteins. This hypothesis would imply that several or even all bacterial species can trigger PBC. Therefore, we investigated whether PBC exhibits immune reactions to non-species-specific bacterial antigens. Yersinia enterocolitica O3 was screened for the presence of proteins that were labeled by immunoblotting using PBC sera. We focused our investigations on a 160-kDa protein, which was further enriched and characterized by partial N-terminal amino acid sequencing. The prevalence of antibodies to this protein was determined by immunoblotting in a variety of diseases. The 160-kDa protein was identified as the beta-subunit of bacterial RNA-polymerase, a highly conserved bacterial protein with a very high degree of sequence identity among all bacterial species. Antibodies to the beta-subunit of bacterial RNA polymerase were specific for this protein. Until now no mammalian protein could be found that cross-reacts with these antibodies. The prevalence of antibodies to the beta-subunit of bacterial RNA polymerase (ARPA) using the protein from Yersinia enterocolitica O3 (serum dilution 1:1000) was: healthy controls (HC, N = 101) 7.9%, primary biliary cirrhosis (PBC, N = 61) 32.8%, autoimmune hepatitis type 1 (AIH, N = 46) 26.1%, alcoholic liver cirrhosis (ALC, N = 44) 9.1%, Crohn's disease (CD, N = 38) 7.9%, ulcerative colitis (UC, N = 24) 8.3%, primary sclerosing cholangitis + UC (PSC/UC, N = 11) 0%, acute yersiniosis (Yers, N = 36) 19.4%, acute infection with Campylobacter jejuni (Camp, N = 10) 0%, acute Q-fever (QF, N = 16) 6.25%, chronic hepatitis C (HCV, N = 39) 7.7%, c-ANCA-positive vasculitis (Vasc, N = 40) 15%, systemic lupus erythematosus (SLE, N = 28) 10.7%, and malaria tropica (MT, N = 24) 16.7%. There was no significant difference between PBC and AIH. The group of autoimmune liver diseases (PBC + AIH, N = 107, 29.9%) differed highly significantly from HC, chronic inflammatory bowel diseases (CD + UC + PSC/UC, N = 73, 6.8%), ALC, and HCV and also differed significantly (P = 0.01) from the group with bacterial and parasitic diseases (Yers + Camp + QF + MT, N = 86,13.95%) and from the group with Vasc + SLE (N = 68,13.2%). Testing of ARPA using the protein from E. coli yielded nearly identical results. In conclusion, an increased prevalence of antibodies to the beta-subunit of bacterial RNA polymerase, a highly conserved non-species-specific bacterial protein, can be found in primary biliary cirrhosis, but also in autoimmune hepatitis type I. These findings do not add an argument for a bacterial trigger of PBC. Rather, they suggest that ARPA belong to the pool of natural antibodies that are up-regulated in autoimmune liver diseases.

Antibodies, Bacterial↗

[Species-specific odor in closely related species: period of development in ontogenesis of house (Mus musculus musculus) and mound-building (Mus spicilegus) mice].

In two-choice odor tests, it was shown that house and mound-building mice began to discriminate the species-specific odor of young at the age of 17 days. The mice investigated the odor of conspecific urine significantly longer than that of heterospecific urine, when the young donors of urine, were 17 days old or older. Thus, the species-specific odor appears in ontogenesis of these species on the 17th day of life.

Animals↗

Species-specific detection of hydrocarbon-utilizing bacteria.

Rapid detection and quantitative assessment of specific microbial species in environmental samples is desirable for monitoring changes in ecosystems and for tracking natural or introduced microbial species during bioremediation of contaminated sites. In the interests of developing rapid tests for hydrocarbon-degrading bacteria, species-specific PCR primer sets have been developed for Pseudomonas aeruginosa, Stentrophomonas (Xanthomonas) maltophilia, and Serratia marsescens. Highly variable regions of the 16S rRNA gene were used to design these primer sets. The amplification products of these primer sets have been verified and validated with hemi-nested PCR and with ligase chain reaction (LCR) techniques, and have been applied to the analyses of environmental water samples. These species-specific primer sets were also chosen to amplify in conjunction with a universal set of PCR primers chosen from highly conserved neighboring sequences in the same gene. These multiplex or competitive PCR procedures enable testing with an internal marker and/or the quantitative estimation of the relative proportion of the microbial community that any one of these species occupies. In addition, this universal PCR primer set amplified the same size amplicon from a wide spectrum of procaryotic and eucaryotic organisms and may have potential in earth biota analyses.

Base Sequence↗

[Typing of ornithosis strains with species-specific antigens].

Species-specific antigens of three ornithosis strains of varying origin were isolated. These antigens were used in the complement-fixation test to study immune sera to different strains. The ornithosis strains studied were represented by three serotypes. To differentiate ornithosis from other chlamidia infections and for the preparation of diagnostic immune sera it is recommended to use species-specific antigens of serotype 3, while for vaccine production the corresponding strains should be used.

Antigens, Viral↗

Detection of human and murine trophoblast-specific antigens and an assessment of their species specificity.

The distribution and species specificity of human and mouse trophoblast-specific surface antigens detected by heterologous anti-mouse ectoplacental cone (anti-EPC) trophoblast and anti-human first-trimester trophoblast plasma membrane (anti-TrPM) antisera were assessed using several immunochemical and immunolabelling assays. The findings with immunofluorescence assays using anti-EPC antiserum on monolayer cultures indicate that the expression of mouse trophoblast-specific antigens is restricted to the major trophoblast components of the mouse placenta, as well as pre- and early post-implantation trophoblast, and does not occur on fetal, amnion or tumour tissues. The anti-EPC antiserum does not cross-react with human chorionic villous trophoblast. The human trophoblast-specific antigens also displayed species specificity. The anti-TrPM antiserum showed no evidence of cross-reactivity with rhesus monkey, rabbit, guinea pig or mouse trophoblast by immunodiffusion, crossed immunoelectrophoresis, immunofluorescence or immunoperoxidase labelling.

Animals↗

Species-specific oligonucleotide probes for five Bifidobacterium species detected in human intestinal microflora.

Portions of the 16S rRNA from closely related species of the genus Bifidobacterium that are found in the human intestinal microflora were sequenced in order to design species-specific oligonucleotide probes. Five oligonucleotide probes ranging from 16 to 19 bases in length and complementary to 16S rRNA sequences from Bifidobacterium adolescentis, B. bifidum, B. breve, B. infantis, and B. longum were synthesized. With crude high-molecular-weight RNA preparations as targets, these probes showed the desired species specificity, even down to a 1-nucleotide difference. For the practical evaluation of these probes, their specificity and sensitivity were tested against seven strains of the same species and 54 strains of heterologous bacteria with fixed whole cells as targets. The probes for B. adolescentis, B. breve, and B. longum showed efficient and specific hybridization. Although the probes for B. bifidum and B. infantis cross-reacted with a few bacterial strains not isolated from humans, these probes showed species specificity for human intestinal bacteria. These 16S rRNA probes should prove valuable for the identification and detection of human intestinal Bifidobacterium species.

Base Sequence↗

Characterization and partial sequencing of species-specific sarcoplasmic polypeptides from commercial hake species by mass spectrometry following two-dimensional electrophoresis.

The Merluccidae family comprises marine species, some of them of high commercial value and others less appreciated, whose commercialization in Europe under the generic name of "hake" is highly remarkable. The potential of proteomics was employed in this study with the aim of achieving the differential characterization of five different hake species: Merluccius merluccius (European hake), M. australis (Southern hake), M. hubbsi (Argentinian hake), M. gayi (Chilean hake), and M. capensis (Cape hake), some of them very closely related. Species-specific polypeptides were observed for the five hake species studied in isoelectric focusing (IEF) and/or two-dimensional electrophoresis (2-DE) high-resolution gels. The peptide mass maps of two polypeptide groups, previously selected by 2-DE analysis as potentially species-specific, were obtained by "in-gel" tryptic digestion, followed by matrix assisted laser desorption/ ionization-time of flight-mass spectrometry (MALDI-TOF-MS). Analysis of group A polypeptides (with pI in the range of 5.0-5.5 and molecular mass of 17 kDa), allowed the differential classification of the hake species into two groups: the East Atlantic coast group and the West Atlantic coast group. Moreover, the peptide mass-maps from the heat-resistant parvalbumin fraction (pI below 4.5; molecular mass <12 kDa) allowed the detection of a peptide characteristic of M. australis not present in the other four hake species tested. A specific 17 kDa protein from M. merluccius was also partially sequenced by nanospray-ion trap-tandem MS, revealing a high homology with rat nucleoside diphosphate kinase A (NDKA). This work opens the way to the application of proteomics to the differential characterization of commercial hake species at the molecular level.

Amino Acid Sequence↗

Cell-specific antigens in chicken erythroid nuclei: species specificity.

Antisera raised to dehistonized chicken reticulocyte chromatin were tested for their cell and species specificity. Quantitative microcomplement fixation and immunohistochemical localization revealed the presence in chromatin of erythroid cell-specific nonhistone protein antigen(s). The antigenic specificity was shown to depend on the association of the antigenic protein(s) with deoxyribonucleic acid (DNA). Although the antisera were exceptionally cell specific, they cross-reacted with erythroid cells of other avian species. The extent of cross-reactivity was found to approximate the phylogenetic distances of the tested avian species. Erythroid cells from fish and amphibians were not reactive. Reconstitution experiments of partially purified chicken reticulocyte chromosomal nonhistone protein antigens with DNAs isolated from several vertebrate species showed that the species specificity of the antigenic complexes is determined principally by the species origin of the nonhistone proteins. Our results show that a cell-specific chromosomal nonhistone protein(s) has undergone evolutionary change and the relative immunological differences are consistent with the accepted phylogenetic distances of the species examined.

Animals↗

Development of novel fluorescence probes that can reliably detect reactive oxygen species and distinguish specific species.

We designed and synthesized 2-[6-(4'-hydroxy)phenoxy-3H-xanthen-3-on-9-yl]benzoic acid (HPF) and 2- [6-(4'-amino)phenoxy-3H-xanthen-3-on-9-yl]benzoic acid (APF) as novel fluorescence probes to detect selectively highly reactive oxygen species (hROS) such as hydroxyl radical (*OH) and reactive intermediates of peroxidase. Although HPF and APF themselves scarcely fluoresced, APF selectively and dose-dependently afforded a strongly fluorescent compound, fluorescein, upon reaction with hROS and hypochlorite ((-)OCl), but not other reactive oxygen species (ROS). HPF similarly afforded fluorescein upon reaction with hROS only. Therefore, not only can hROS be differentiated from hydrogen peroxide (H(2)O(2)), nitric oxide (NO), and superoxide (O2*-) by using HPF or APF alone, but (-)OCl can also be specifically detected by using HPF and APF together. Furthermore, we applied HPF and APF to living cells and found that HPF and APF were resistant to light-induced autoxidation, unlike 2',7'-dichlorodihydrofluorescein, and for the first time we could visualize (-)OCl generated in stimulated neutrophils. HPF and APF should be useful as tools to study the roles of hROS and (-)OCl in many biological and chemical applications.

Carcinoma, Hepatocellular↗

Design of species-specific primers to identify 13 species of Clostridium harbored in human intestinal tracts.

The genus Clostridium is dominant in human intestinal tracts and plays an important role in human health. We designed species-specific primers to identify 13 species of Clostridium (C. perfringens, C. paraputrificum, C. bifermentans, C. difficile, C. clostridiiforme, C. nexile, C. sphenoides, C. indolis, C. ramosum, C. cocleatum, C. butyricum, C. sordellii, and C. innocuum) easily and rapidly. The PCR annealing temperature was set at a uniform 60 C for application to all strains at the same time. To confirm the specificities of these primers, 85 intestinal bacteria in total, including type strains, reference strains, and isolates were used. Ten primers (including those for C. perfringens to C. cocleatum) indicated high specificities. Although there were some cross-reactions with the other three primers, the target species were distinguishable from other bacteria by the different sizes of PCR products.

Clostridium↗

Production of specific antibodies to contractile proteins, and their use in immunofluorescence microscopy. II. Species-specific and species-non-specific antibodies to smooth and striated chicken muscle actin.

The injection of rabbits with insoluble or soluble G-actin from chicken smooth or striated muscle will produce antibodies that are equally reactive, and species and tissue non-specific in immunoprecipitation, immunofluorescence and actin-activated Mg2+-ATPase inhibition tests. These antibodies have been used for the identification of actin-containing fibrils in a variety of tissues. When G-actins from chicken smooth or striated muscle are immobilized by chemical linkage to Affi-Gel 702 microbeads, their immunogenicity is increased, but the antibodies obtained against them are species-specific and will only react with actin and actin-containing structures from chicken and are therefore limited in use. It is concluded from this work that insoluble G-actin is the preferable immunogen to obtain precipitating antibodies for wide use.

Actins↗

Molecular studies on the aerolysin gene of Aeromonas species and discovery of a species-specific probe for Aeromonas trota species nova.

A large group of aeromonads and other enteric microorganisms were assayed for the presence of the aerolysin gene with use of DNA-DNA hybridization. Two DNA fragments corresponding to the regulatory region (aerC) and the structural gene (aerA) were used as probes for the detection of the aerolysin gene in these strains. Sequences corresponding to the aerolysin structural gene were widespread among Aeromonas isolates. In contrast, the aerC probe was much more selective, and sequences corresponding to the aerC region were detected in only a small subset of strains. Concurrent studies using numerical taxonomy and DNA hybridization with the aerC probe on a larger set of strains led to the identification of a distinct cluster of 14 presumed atypical Aeromonas sobria strains. These strains have recently been grouped into a new species designated Aeromonas trota. Hence, the DNA fragment aerC used in the study is a species-specific gene probe for A. trota. The ability of the aerC probe to detect strains belonging to a single species suggests that there is selection pressure to maintain the clonality of this species. These results have important implications with respect to the evolution of "pathogenic profiles" among these medically important bacteria.

Aeromonas↗

Streptokinases produced by pathogenic group C streptococci demonstrate species-specific plasminogen activation.

The species specificities of plasminogen activation and binding of plasmin by pathogenic group C streptococci isolated from humans, horses, and pigs were examined. Of 56 streptococcal isolates, 52 elaborated plasminogen activator activity and 49 of these had specificity for plasminogen of the homologous host. Analysis of supernatants from 13 isolates indicated that the plasminogen activator activity resulted from secreted streptokinases. These 13 streptokinases were antigenically related and bound all three plasminogens, indicating that the binding recognition sites were conserved despite the observed species-specific activation. In addition, all group C isolates tested demonstrated surface receptors that bound human, equine, and porcine plasmin. Species-specific plasminogen activation may be an early step in events resulting in infection and may account for the species preference of certain streptococci.

Animals↗