Characterization of a wild-type strain of Francisella tularensis isolated from a cat.

Francisella tularensis type A is the primary cause of tularemia in animals and humans in North America. The majority of research on F. tularensis has been done with the attenuated live vaccine strain (LVS), which is a type B, but very few wild-type F. tularensis strains have been characterized. A gram-negative coccobacillus that was isolated in pure culture from the lungs of a cat that died after being lost for 5 days was received for identification at the Virginia-Maryland Regional College of Veterinary Medicine Teaching hospital. The isolate (strain TI0902) was not identified (or was misidentified) by commercial identification systems; however, it was identified as F. tularensis subspecies tularensis (type A) by sequencing a portion of the 16S ribosomal RNA gene. Furthermore, repetitive extragenic palindromic sequences-polymerase chain reaction amplified a 4-kb DNA fragment from TI0902 that was characteristic of F. tularensis type A but not type B. The electrophoretic profile of the lipopolysaccharide of strain TI0902 was identical to that of the LVS by Western blotting with antiserum to LVS. The protein-enriched outer membrane of strain TI0902 contained 6-8 proteins, which were similar in molecular size to those from the LVS. Electron microscopy of negatively stained and alcian blue-stained LVS and TI0902 cells showed that both strains were coccobacillary in shape and may be encapsulated. However, after mouse challenge, the TI0902 strain was clearly more virulent than the LVS strain. Results of this study indicate that the genotype and phenotype of wild-type F. tularensis type A strain TI0902 is similar, but not identical, to that of the LVS strain. Further studies will help determine whether pathogenesis and host-pathogen interactions are also similar between the 2 strains.     

Pancreatic islet fibrosis in rock hyraxes (Procavia capensis), Part 1: Case histories, clinical pathology, and epizootiology.

Two adult female rock hyraxes (Procavia capensis) at the Dallas Zoo were confirmed with spontaneous diabetes mellitus from 1997-2000, whereas a third animal with a similar clinical presentation never became hyperglycemic. The pancreas in all three animals showed pancreatic islet fibrosis (PIF). Retrospective examination of medical records for rock hyraxes acquired by this collection or born into it from 1991-2002 identified eight more animals affected with PIE All affected animals, including three males and eight females, were 1-7 yr of age and presented either with vague clinical signs of soft feces and rough hair coat or were acutely moribund or dead. Clinical pathology data was available for seven of the animals before onset of overt clinical signs and revealed inappropriate hyperglycemia in six, as well as elevated serum concentrations of creatine phosphokinase, amylase, and lipase in all seven animals. Pedigree evaluation did not support a familial pattern for PIE Review of the histopathology findings from nine other zoologic collections with rock hyrax deaths during the study period identified six institutions with 12 additional cases genetically unrelated to the incident collection. Histopathology and viral serology did not support an infectious cause. Analysis of serum anti-islet and anti-insulin antibodies did not suggest autoimmune disease, and none of the animals had known exposure to toxic substances. Limited nutritional analyses did not support a nutritional basis for the condition, and the cause for PIF remains unknown.     

Competition Effects Among Isolates of Fusarium culmorum Differing in Aggressiveness and Mycotoxin Production on Heads of Winter Rye

Fusarium culmorum is a phytopathogenic, toxigenic fungus causing seedling diseases, foot rot and head blight of cereals. For estimating competition effects in mixtures of two single-spore isolates, two winter rye single crosses were tested with either four isolates individually or four 1 : 1 mixtures of the same isolates in six field environments. Two isolates (FC46, FC64) were highly aggressive deoxynivalenol (DON) and 3-acetyl DON-producers, the other two (FC30, FC71) were medium aggressive nivalenol-producers. Rye heads were inoculated during flowering with conidia of pairs of isolates expressing similar (FC46 + FC64, FC30 + FC71) or contrary (FC46 + FC71, FC30 + FC64) levels of aggressiveness and similar or different concentrations and chemotypes of mycotoxins, respectively. Head blight rating and yield components relative to the non-inoculated plots were recorded as aggressiveness traits. Additionally, mycotoxin concentrations were measured in the rye grain. Random pathogen samples were re-isolated from heads at the onset of symptom development and analysed by molecular markers (RAPD–PCR) in one environment. Aggressiveness of the isolate mixtures was significantly lower than that of the isolates applied individually on both rye genotypes. Similarly, mycotoxin concentrations were significantly lower in the mixtures in seven out of eleven comparisons. Among the re-isolates, the component genotypes of a mixture significantly deviated from the inoculated 1 : 1 ratio when a particular isolate (FC46) was present in the mixture. This isolate displayed a superior competitive ability irrespective of the aggressiveness or mycotoxin profile of the mixing partner illustrating that pathogenic fitness is caused by additional factors that have not, as yet, been identified.     

Analysis of Mycosporine-Like Amino Acids in Selected Algae and Cyanobacteria by Hydrophilic Interaction Liquid Chromatography and a Novel MAA from the Red Alga Catenella repens

Mycosporine-like amino acids (MAAs), a group of small secondary metabolites found in algae, cyanobacteria, lichens and fungi, have become ecologically and pharmacologically relevant because of their pronounced UV-absorbing and photo-protective potential. Their analytical characterization is generally achieved by reversed phase HPLC and the compounds are often quantified based on molar extinction coefficients. As an alternative approach, in our study a fully validated hydrophilic interaction liquid chromatography (HILIC) method is presented. It enables the precise quantification of several analytes with adequate retention times in a single run, and can be coupled directly to MS. Excellent linear correlation coefficients (R² > 0.9991) were obtained, with limit of detection (LOD) values ranging from 0.16 to 0.43 µg/mL. Furthermore, the assay was found to be accurate (recovery rates from 89.8% to 104.1%) and precise (intra-day precision: 5.6%, inter-day precision ≤6.6%). Several algae were assayed for their content of known MAAs like porphyra-334, shinorine, and palythine. Liquid chromatography-mass spectrometry (LC-MS) data indicated a novel compound in some of them, which could be isolated from the marine species Catenella repens and structurally elucidated by nuclear magnetic resonance spectroscopy (NMR) as (E)-3-hydroxy-2-((5-hydroxy-5-(hydroxymethyl)-2-methoxy-3-((2-sulfoethyl)amino)cyclohex-2-en-1-ylidene)amino) propanoic acid, a novel MAA called catenelline.     

Influence of larval and juvenile life history on age at first maturity in two tropical amphidromous fish species

This study investigates how age at first maturity of two tropical amphidromous species Sicyopterus lagocephalus (Pallas, 1770) and Cotylopus acutipinnis (Guichenot, 1863) varies in relation to their larval and juvenile life history. Reproductive stage was estimated based on histological observation of ovaries of more than 200 females of each species caught monthly over 1 year. The age of fish was estimated by interpreting the daily increments deposited on otoliths during the oceanic larval phase, and the juvenile phase in the river. The age at first maturity was approximately 9 months for S. lagocephalus and 7 months for C. acutipinnis, corresponding to approximately 70–130 and 90–130 days after they returned to freshwater respectively. For both species, the time spent in freshwater before maturity was significantly influenced by the duration of the pelagic larval stage at sea (PLD) and the season of return in freshwater. Individuals with a long PLD, or returning in freshwater during the warmer season, maturated faster once in freshwater. This reproductive advantage may minimise the risk of extirpation due to catastrophic events at each generation and thus probably benefits amphidromous species living in very unpredictable tropical rivers.     

Evidence for genetic distinction among sympatric ecotypes of Arctic char (Salvelinus alpinus) in south‐western Alaskan lakes

Resource polymorphism may play an important role in the process of speciation. The Arctic char (Salvelinus alpinus) exhibits great phenotypic and genetic diversity across its range, making it an ideal species for studies of resource polymorphism and divergence. Here, we investigated genetic variation at 11 microsatellite loci among 287 Arctic char from five isolated yet proximate postglacial lakes in south‐western Alaska that were previously examined for resource polymorphism. Significant differences in pairwise F_ST were detected among all lakes (range from 0.05 to 0.28, all P < 0.02). In one lake (Lower Tazimina Lake), we found evidence for two genetic groups of char and for significant differences in the distribution of microsatellite variability among at least two of the three previously described body size morphotypes (‘large’‐, ‘medium’‐, and ‘small’‐bodied char; maximum F_ST = 0.09; differences in admixture proportions). We also found a significant association between genetic admixture proportions and gill raker counts among body size morphs (r = ?0.73, P < 0.001). Our data represent the first record of genetically distinct sympatric morphs of Arctic char in Alaska and provide further evidence that differences in morphology associated with feeding (gill rakers) and growth trajectories reflect niche diversification and promote genetic divergence in Holarctic populations of Arctic char.