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Reclassification of Paenibacillus larvae subsp. pulvifaciens and Paenibacillus larvae subsp. larvae as Paenibacillus larvae without subspecies differentiation.

A polyphasic taxonomic study of the two subspecies of Paenibacillus larvae, Paenibacillus larvae subsp. larvae and Paenibacillus larvae subsp. pulvifaciens, supported the reclassification of the subspecies into one species, Paenibacillus larvae, without subspecies separation. Our conclusions are based on the analysis of six reference strains of P. larvae subsp. pulvifaciens and three reference strains and 44 field isolates of P. larvae. subsp. larvae. The latter originated from brood or honey of clinically diseased honey bee colonies or from honey of both clinically diseased and asymptomatic colonies from Sweden, Finland and Germany. Colony and spore morphology, as well as the metabolism of mannitol and salicin, did not allow a clear identification of the two subspecies and SDS-PAGE of whole-cell proteins did not support the subspecies differentiation. For genomic fingerprinting, repetitive element-PCR fingerprinting using ERIC primers and PFGE of bacterial DNA were performed. The latter method is a high-resolution DNA fingerprinting method proven to be superior to most other methods for biochemical and molecular typing and has not previously been used to characterize P. larvae. ERIC-PCR identified four different genotypes, while PFGE revealed two main clusters. One cluster included most of the P. larvae subsp. larvae field isolates, as well as all P. larvae subsp. pulvifaciens reference strains. The other cluster comprised the pigmented variants of P. larvae subsp. larvae. 16S rRNA gene sequences were determined for some strains. Finally, exposure bioassays demonstrated that reference strains of P. larvae subsp. pulvifaciens were pathogenic for honey bee larvae, producing symptoms similar to reference strains of P. larvae subsp. larvae. In comparison with the type strain for P. larvae subsp. larvae, ATCC 9545T, the P. larvae subsp. pulvifaciens strains tested were even more virulent, since they showed a shorter LT100. An emended description of the species is given.

Animals↗

Pleomorphic leiomyosarcoma of the adrenal gland with osteoclast-like giant cells.

Pleomorphic leiomyosarcoma (PLMS) of the adrenal gland is a rare tumor in an unusual location. A primary PLMS of the left adrenal gland is reported in a 59-yr-old Mexican woman who presented progressive flank pain and weight loss. The tumor measured 16 cm in diameter, showed markedly pleomorphic and osteoclast-like giant cells, necrosis, and high mitotic activity (average 15 per 10 high-power fields). The phenotype was supported by light microscopy and corroborated by immunohistochemistry. The neoplastic cells were strongly positive for muscle-specific actin, desmin, vimentin, and p53. They were negative for CD34, HMB45, estrogen receptors, and S-100 protein. The percentage of Ki-67 positive neoplastic cells was 7.6%. DNA content analysis by flow cytometry showed that tumor was diploid, with a high level of apoptosis. Extra-adrenal primary sites of origin were clinically excluded. The patient developed local recurrence and liver metastases 12 mo after initial treatment. She then received adjuvant chemotherapy and radiotherapy and the metastasis was resected. Twenty-four months later, she is alive with no evidence of disease. This is the second case of adrenal PLMS reported. This case exhibited a high histologic grade, aggressive behavior, and p53 overexpression, but diploid DNA content.

Adrenal Gland Neoplasms↗

[Genetic analysis of osteoporosis susceptibility: future perspectives on genome wide analysis].

Genetic susceptibility of osteoporosis has been analyzed mainly via sib-pair analysis and association study on candidate genes, however these approaches might have substantial limitation. Recently proposed genome wide analysis of association using high density polymorphic markers has not been established on this field. Preferable phenotypes and suitable subjects have to be defined as well as for tag-SNP selection and the method of multi-step screening.

Genetic Predisposition to Disease↗

Members of the novel WC1 gene family are differentially expressed on subsets of bovine CD4-CD8- gamma delta T lymphocytes.

CD4-CD8- gamma delta T cells of ruminants uniquely express a 220-kDa surface Ag recognized by several mAbs clustered as WC1. We recently reported the isolation of a cDNA clone encoding a WC1 Ag. Southern blotting suggested that the bovine genome contains multiple sequences highly related to the isolated WC1 cDNA. Here, we demonstrate that some of the clustered WC1 mAbs stain predominantly nonoverlapping subsets of bovine CD4-CD8- gamma delta T cells. By the isolation of two additional cDNA clones encoding molecules highly related to the original WC1 Ag, we provide a molecular basis for this phenomenon. Cells transfected with cDNAs encoding individual WC1 Ags are differentially recognized by various WC1 mAbs. Thus, expression of members of the WC1 gene family divides bovine CD4-CD8- gamma delta T cells into phenotypical subsets. Field inversion gel electrophoresis revealed that all WC1 genes map to a single, large (> 1 Mbp) Notl fragment. Although the function of WC1 remains unknown, it likely involves interaction with ligands that originate from a similarly complex genetic system.

Amino Acid Sequence↗

Genetic and phenotypic correlations between feather pecking and open-field response in laying hens at two different ages.

The object of this research was to study the relationship between feather pecking and open-field activity in laying hens at two different ages. A population of 550 birds of a laying hen cross was subjected to an open-field test at 5 and 29 weeks of age and to a social feather pecking test at 6 and 30 weeks of age. Factor analysis was used to identify underlying factors for each test: pecking behavior (social test) and open-field activity (open-field test). In young birds, a positive phenotypic correlation of 0.24 was found between high open-field activity and high levels of pecking behavior (ground pecking, preening, gentle feather pecking, and wall pecking). In adults, a similar genetic correlation of 0.62 was found. At adult age, the factor pecking behavior consisted mainly of gentle and severe feather pecking. Between ages, a strong, negative genetic correlation of -0.65 was found between open-field activity at young age and pecking behavior at adult age, indicating that open-field activity levels in young birds may predict pecking behavior in adult hens.

Aggression↗

[Phenotypic characterization of equine Dermatophilus congolensis field isolates].

In 1993 and 1994 a highly increased occurrence of equine dermatophilosis was observed, and a study was initiated to determine phenotypic heterogeneity among 120 clinical isolates using biochemistry, antibiotic resistance profiles, membrane protein profiles and Western blotting. The biochemical examinations contained 1% equine serum in medium. Moreover, the API ZYM-test from bioMérieux was used. The biochemical reactions were suited to identify Dermatophilus congolensis but did not allow a differentiation among the various isolates. Antibiotic resistance in one or more isolates was observed against polymyxin B, enrofloxacin, oxacillin, neomycin and trimethoprim-sulfamethoxazol. All isolates were sensitive penicillin G, ampicillin, streptomycin, gentamicin, lincomycin, erythromycin, tetracycline, oxytetracycline, bacitracin and ceftiofur. The evaluation of silver-stained and immuno-stained membrane protein profiles showed minor differences among several isolates. In total, all isolates appeared to be closely related and the minor differences observed did not correlate with the geographic origin of the respective isolates.

Actinomycetales↗

Isolates of infectious bursal disease virus from India are of very virulent phenotype.

Four Indian field isolates, a classical virulent and an attenuated vaccine strains of Infectious bursal disease virus (IBDV) have been characterized by sequence analysis of part of the VP1 gene (from nucleotide 1538-1979) comprising one of viral RNA dependent RNA polymerase motifs. Sequence alignment of these viruses with reported viruses of other countries revealed Indian IBDV field isolates to be 100% similar to very virulent Japanese (OKYM), European (UK661) and Bangladesh (BD3/99) IBD viruses at amino acid level, whereas they had 0.2-0.9% divergence at nucleotide level. Out of the total 24 nucleotide changes found in the Indian field isolates, as well as reported very virulent viruses, only one resulted in amino acid change S-P at 562 position. The Indian field isolates displayed nucleotide divergence of 10.6-11.6% and amino acid divergence of 2.8-3.5% from the classical virulent and attenuated vaccine strains. The RNA dependent RNA polymerase motif from amino acid 528-541, present in the sequence analyzed, was conserved among all the viruses, irrespective of pathotype and serotype. In the phylogenetic tree, based on nucleotide sequence, Indian field viruses were grouped with reported very virulent viruses in one lineage whereas, classical virulent, attenuated vaccine and serotype 2 strains formed part of the second lineage. But in the phylogenetic tree based on amino acid sequence alignment, the serotype 2 strain OH grouped with Indian field isolates and reported very virulent viruses in one lineage and classical virulent and attenuated vaccine strains formed the second lineage.

Amino Acid Sequence↗

Prevalence of cpb2, encoding beta2 toxin, in Clostridium perfringens field isolates: correlation of genotype with phenotype.

Beta2 toxin, encoded by the cpb2 gene, has been implicated in the pathogenesis of porcine, equine and bovine enteritis by type A Clostridium perfringens. By incorporating primers to cpb2 into a multiplex genotyping PCR, we screened 3270 field isolates of C. perfringens. Of these, 37.2% were PCR positive for the cpb2 gene. The majority of isolates from cases of porcine enteritis were positive for cpb2 (>85%), and this was even more true for C. perfringens isolated from cases of porcine neonatal enteritis (91.8%). In contrast, isolates from normal pigs only contained cpb2 in 11.1% of cases. The correlation between enteritis in other animal species and the presence of cpb2 was not so strong. cpb2 was found in 21.4% of C. perfringens isolates from cattle with enteritis, and in 47.3% of isolates from calves with enteritis or abomastitis. The prevalence of cpb2 varied with genotype, with type A isolates being positive for this gene in 35.1% of cases. Furthermore, enterotoxigenic type D or type E strains almost always carried cpb2. We cloned a 6xHIS-tagged beta2 (HIS-beta2) and used this protein to raise antiserum against beta2. Culture supernatants from 68 cpb2-positive and 13 cpb2-negative strains were tested for the presence of beta2 by Western blotting. In cpb2-positive isolates of porcine origin, beta2 was almost always detected (96.9%). However, in cpb2-positive isolates from other animal species, only 50.0% expressed beta2 protein. The high rate of cpb2-positivity among strains from neonatal pigs with enteritis and the high correlation of genotype with phenotype, supports the contention that beta2 toxin plays a role in the pathogenesis of these infections. However, it may be important to consider the use of an additional method for the detection of beta2 toxin in non-porcine cpb2-positive isolates when making claims about the role of beta2 in enteritis in non-porcine species.

Animals↗

Phenotyping of erythrocyte acid phosphatase and esterase D by high field strength isoelectric focusing on cellulose acetate membrane.

Phenotyping of erythrocyte acid phosphatase (EAP) and esterase D (ESD) by cellulose acetate membrane isoelectric focusing (CAM-IEF) under a nonequilibrium condition is described. In an attempt to improve the method of CAM-IEF, we shortened the electrode distance to provide a higher field strength at a given (low) voltage. Various carrier ampholytes for EAP typing and various chemical separators for ESD typing were also tested. Good separations were obtained after 30 min IEF for EAP typing and 25 min for ESD typing. When applied to blood stains and stored for various periods at room temperature, the stains up to 8 months old could still be phenotyped for EAP and those up to 4 weeks old for ESD. CAM-IEF is suitable for routine forensic work of EAP and ESD phenotyping.

Acid Phosphatase↗

Investigation of specific substitutions in virulence genes characterizing phenotypic groups of low-virulence field strains of Listeria monocytogenes.

Several models have shown that virulence varies from one strain of Listeria monocytogenes to another, but little is known about the cause of low virulence. Twenty-six field L. monocytogenes strains were shown to be of low virulence in a plaque-forming assay and in a subcutaneous inoculation test in mice. Using the results of cell infection assays and phospholipase activities, the low-virulence strains were assigned to one of four groups by cluster analysis and then virulence-related genes were sequenced. Group I included 11 strains that did not enter cells and had no phospholipase activity. These strains exhibited a mutated PrfA; eight strains had a single amino acid substitution, PrfAK220T, and the other three had a truncated PrfA, PrfADelta174-237. These genetic modifications could explain the low virulence of group I strains, since mutated PrfA proteins were inactive. Group II and III strains entered cells but did not form plaques. Group II strains had low phosphatidylcholine phospholipase C activity, whereas group III strains had low phosphatidylinositol phospholipase C activity. Several substitutions were observed for five out of six group III strains in the plcA gene and for one out of three group II strains in the plcB gene. Group IV strains poorly colonized spleens of mice and were practically indistinguishable from fully virulent strains on the basis of the above-mentioned in vitro criteria. These results demonstrate a relationship between the phenotypic classification and the genotypic modifications for at least group I and III strains and suggest a common evolution of these strains within a group.

Amino Acid Substitution↗

Temporal variation of RAPD-PCR phenotype composition of the grain aphid Sitobion avenae (Hemiptera: Aphididae) on wheat: the role of hydroxamic acids.

Hydroxamic acids (Hx) contained in wheat are active mutagens which play an important role in the defence of the plant against aphids. Random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) dominant markers were used to assess genetic variability in the aphid Sitobion avenae (Fabricius) in relation to hydroxamic acid levels in their host-plants. Colonies of aphids belonging to a single RAPD-PCR profile were grown on different host-plants differing in their Hx content under greenhouse conditions. The RAPD-PCR phenotypic pattern showed the appearance of two new RAPD-PCR variants after four to five generations of exposure to wheat cv. Chagual (high Hx levels), one after exposure to wheat cv. Huayún (low Hx levels), and none after exposure to oat (lacking Hx). Differential appearance of new RAPD-PCR aphid phenotypes also occurred on field-grown wheat. While the overall phenotypic 'richness' diminished during the season, the number of RAPD-PCR phenotypes decreased on cv. Huayún and increased on cv. Chagual. The preferential appearance in the field and in the greenhouse of new RAPD-PCR phenotypes of S. avenae on cv. Chagual is discussed on the basis of mutagenesis induced by hydroxamic acids and by the products of their transformation within the aphid. Aphid abundance is interpreted in terms of antixenosis and antibiosis by hydroxamic acids. The appearance on cv. Chagual of phenotypes first detected on cv. Huayún was accounted for by intercrop migrations.

Animals↗

Selection practices and genetic and phenotypic trends estimated from Nebraska SPF swine field records.

Records collected during 1971 through 1979 from 101,606 pigs raised in 18 herds that were members of the Nebraska SPF Swine Accrediting Association were evaluated for phenotypic trends and predicted and observed genetic trends. Traits considered were backfat at 100 kg (BF) and weight at 140 d of age (WT). Phenotypic change on average was beneficial for BF (-.05 mm/yr) but undesirable for WT (-.2 kg/yr). However, the average observed genetic trend was nil for BF and .6 kg/yr for WT. An average, predicted response based on observed selection differentials and estimates of within herd-sex genetic parameters was in good agreement with observed response for BF, but was three times higher for WT.

Adipose Tissue↗

[Behaviour phenotypes].

INTRODUCTION: In the same way as the genetic basis of disease and developmental disorders with behavioural and intellectual features are being discovered, the opportunity of studying how genes may affect conduct is also developing in both dysfunctional and normal situations. DEVELOPMENT: The most useful models for studying this problem are the genetic disorders in which the genetic defect has been discovered or is being traced. In parallel with advances in genetics, the behaviour patterns found in each disorder are also being defined and are known as behaviour phenotypes. CONCLUSIONS: In this article we aim to illustrate information currently available in the field of behaviour phenotypes. The behaviour and intellectual characteristics of the following syndromes are therefore described: fragile X, premutation of the fragile X syndrome, velocardiofacial, Prader Willi, Rett, Angelman and Williams.

Angelman Syndrome↗

Macrorestriction analysis of clinical and environmental isolates of Sporothrix schenckii.

Sporothrix schenckii causes sporotrichosis, a disease that most commonly presents as a subacute or chronic skin infection. An unusually high incidence of clinical cases of sporotrichosis occurred in the southwest of Western Australia over the last 5 years. Anecdotal accounts from patients implicated contact with hay prior to infection. Isolates of S. schenckii from hay and clinical cases were investigated by traditional phenotypic methods and pulsed-field gel electrophoresis (PFGE). The phenotypic evaluation separated S. schenckii from Ophiostoma spp. A DNA macrorestriction method using SfiI and NotI macrorestriction digestion by PFGE was developed to investigate the epidemiological connections. BioNumerics software was used to analyze the results. DNA macrorestriction digestion patterns for the recent Western Australian clinical isolates and four hay isolates were indistinguishable. Eastern state clinical isolates, national Quality Assurance Program isolates, and other environmental isolates gave different macrorestriction patterns. Clinical isolates from the southwest of Western Australia collected in the 1980s and 1990s were also characterized using PFGE. The patterns generated were indistinguishable from those of the recent clinical isolates. PFGE showed that the dominant strain of S. schenckii causing sporotrichosis in Western Australia is present in hay, has caused sporotrichosis for at least 15 years, and is a different strain from the strains found in other parts of Australia.

Australia↗

Phenotypic diversity is the cornerstone of evolution: variation in cortical field size within short-tailed opossums.

Natural selection operates on phenotypic variation that exists within a population. Variable aspects of cortical organization, such as the size and connections of a cortical field, can generate differences in behavior, which is a target of natural selection. Yet studies pertaining to within-species variability in cortical organization are limited. In the present investigation, we examined variation in brain size, cortical sheet size, and primary sensory cortical field sizes in the adult short-tailed opossum (Monodelphis domestica). Within individuals, we found no significant difference between the right and left hemispheres in the overall size of the dorsolateral cortex or in primary cortical field sizes. Between individuals, we found relatively little intraspecies variation in brain weight, brain volume, and cortical sheet area for the dorsolateral neocortex and pyriform cortex; however, we observed a large degree of variability in body weight and primary sensory cortical field size, as defined by myeloarchitecture. Further, we found that the size of each cortical field correlated with the size of the other cortical fields as well as with the total size of the dorsolateral cortex. Here we discuss the possible sources of variation and examine the relationship between cortical field size and sensory processing abilities and behaviors across species. Since behavior is the target of natural selection, variation in cortical field size across individuals may supply the raw material necessary for cortical field evolution.

Afferent Pathways↗

LRTae: improving statistical power for genetic association with case/control data when phenotype and/or genotype misclassification errors are present.

BACKGROUND: In the field of statistical genetics, phenotype and genotype misclassification errors can substantially reduce power to detect association with genetic case/control studies. Misclassification also can bias population frequency parameters such as genotype, haplotype, or multi-locus genotype frequencies. These problems are of particular concern in case/control designs because, short of repeated sampling, there is no way to detect misclassification errors. We developed a double-sampling procedure for case/control genetic association using a likelihood ratio test framework. Different approaches have been proposed to deal with misclassification errors. We have chosen the likelihood framework because of the ease with which misclassification probabilities may be incorporated into in the statistical framework and hypothesis testing. The statistic is called the Likelihood Ratio Test allowing for errors (LRTae) and is freely available via software download. RESULTS: We applied our procedure to 10,000 replicates of simulated case/control data in which we introduced phenotype misclassification errors. The phenotype considered is Ankylosing Spondylitis (AS). The LRTae method power was always greater than LRTstd power for the significance levels considered (5%, 1%, 0.1%, 0.01%). Power gains for the LRTae method over the LRTstd method increased as the significance level became more stringent. Multi-locus genotype frequency estimates using LRTae method were more accurate than estimates using LRTstd method. CONCLUSION: The LRTae method can be applied to single-locus genotypes, multi-locus genotypes, or multi-locus haplotypes in a case/control framework and can be more powerful to detect association in case/control studies when both genotype and/or phenotype errors are present. Furthermore, the LRTae method provides asymptotically unbiased estimates of case and control genotype frequencies, as well as estimates of phenotype and/or genotype misclassification rates.

Case-Control Studies↗

Phenotypic and allozyme variation in Mediterranean and desert populations of wild barley, Hordeum spontaneum Koch.

Populations of wild barley, Hordeum spontaneum Koch, were collected in two distinct climatic regions, desert and Mediterranean. Plants from five desert and five Mediterranean populations were compared and contrasted for extent and structure of phenotypic variation. These same 10 and one other population from each region were analyzed for allozyme variation. In a field trial of phenotypic diversity, two phenological and 14 morphological traits were examined. Study of allozyme variation was performed using eight enzyme systems encoding for 13 loci. Plants from the desert and Mediterranean regions were significantly different in seven of 16 phenotypic traits, exhibited a high (30%) interregional component of phenotypic variation, and showed a high degree of segregation on a principal component scattergram indicating ecotypic differentiation. Mediterranean populations were twice as variable as desert populations in reproductive growth parameters (stem and spike length) and grain filling (spikelet weight), but half as variable for onset of reproduction. The extent and structure of phenotypic and allozyme variation did not match. The Mediterranean and desert populations did not differ in amount of allozyme variation as estimated by mean number of alleles per locus, effective number of alleles, polymorphism, and gene diversity (n(a), n(e), P, and H(e)), did not segregate on the basis of population genetic distances, and exhibited a low proportion of interregion allozyme diversity (2%). No effect of selection on allozyme distribution was detected. Our results suggest that the adaptation of plants originating from desert and Mediterranean environments is reflected in phenotypic but not in allozyme variation.

Desert Climate↗

Genetic and phenotypic traits of streptomycetes used to characterize antibiotic activities of field-collected microbes.

Although antibiotic production may contribute significantly to microbial fitness, there is limited information on the ecology of antibiotic-producing microbial populations in soil. Indeed, quantitative information on the variation in frequency and intensity of specific antibiotic inhibitory and resistance abilities within soil microbial communities is lacking. Among the streptomycetes, antibiotic production is highly variable and resistance to antibiotics is highly specific to individual microbial strains. The objective of this work was to genetically and phenotypically characterize a reference collection of streptomycetes for use in distinguishing inhibition and resistance phenotypes of field-collected microbes. Specifically, we examined inhibition and resistance abilities of all isolates in all possible pairwise combinations, genetic relatedness using BOX-PCR and 16S rDNA sequence analyses, nutrient utilization profiles, and antibiotic induction among all possible three-way combinations of isolates. Each streptomycete isolate possessed a unique set of phenotypic and genetic characteristics. However, there was little correspondence between phenotypic and genetic traits. This collection of reference isolates provides the potential for distinguishing 1024 inhibition and resistance phenotypes in field-collected microbes. Relationships between the genetic and phenotypic characteristics examined may provide preliminary insight into the distinct strategies that microbes use in optimizing their fitness in natural environments.

Anti-Bacterial Agents↗