Karyotype analysis of Leishmania species and its use in classification and clinical diagnosis

Chromosomes of four species of Leishmania represented by ten different geographic isolates were analyzed by pulsed field gradient gel electrophoresis (PFG) to assess chromosome stability in these parasitic protozoans. Among different geographic isolates of the same subspecies, more than two-thirds of chromosomes had similar sizes, ethidium bromide staining intensities, and locations of alpha,beta-tubulin genes. However, among New World Leishmania, members of different species or subspecies have fewer than one-third of their chromosomes in common. Therefore, PFG karyotypes of Leishmania exhibit intraspecific variability similar to that reported for other parasitic protozoans. The greater similarities of the karyotypes of members of the same Leishmania subspecies may indicate that they represent valid taxa. These similarities also allowed the use of PFG in clinical diagnosis for rapid and accurate typing of patient isolates.     

A virus in a fungus in a plant: three-way symbiosis required for thermal tolerance

A mutualistic association between a fungal endophyte and a tropical panic grass allows both organisms to grow at high soil temperatures. We characterized a virus from this fungus that is involved in the mutualistic interaction. Fungal isolates cured of the virus are unable to confer heat tolerance, but heat tolerance is restored after the virus is reintroduced. The virus-infected fungus confers heat tolerance not only to its native monocot host but also to a eudicot host, which suggests that the underlying mechanism involves pathways conserved between these two groups of plants.     

Origin and evolution of prebiotic organic matter as inferred from the Tagish Lake meteorite

The complex suite of organic materials in carbonaceous chondrite meteorites probably originally formed in the interstellar medium and/or the solar protoplanetary disk, but was subsequently modified in the meteorites' asteroidal parent bodies. The mechanisms of formation and modification are still very poorly understood. We carried out a systematic study of variations in the mineralogy, petrology, and soluble and insoluble organic matter in distinct fragments of the Tagish Lake meteorite. The variations correlate with indicators of parent body aqueous alteration. At least some molecules of prebiotic importance formed during the alteration.     

Book reviews

Marilyn Holly is an Associate Professor of Philosophy at the University of Florida. Her special interests include political philosophy, ethics and ecology, Jungian thought, and Native American spirituality in its relation to the environment. She is currently at work on a monograph-length piece on the latter topic. She has taught an undergraduate course in ethics and ecology, and plans to do so again. The following essay emerged from her search for a suitable text for such a course.     

A rapid spectrofluorometric screening method for enrofloxacin in chicken muscle

A simple spectrofluorometric method was developed for screening enrofloxacin (ENRO) in chicken muscle. A single-step extraction with acidic acetonitrile gave the best results without further cleanup. Following centrifugation the supernatants were excited at 324 nm and the emission was measured at 442 nm. Using this procedure, 18 chicken breast samples from 3 producers were tested. The results showed background signal levels significantly lower than those corresponding to 300 microg/kg ENRO, the FDA approved tolerance level. Statistical treatment of these data established a threshold which can be used in subsequent screening of ENRO at the tolerance level. The calibration curve revealed a satisfactory linear relationship (R(2) = 0.9991) in a range of 0-700 microg/kg ENRO in fortified chicken breast. ENRO-incurred samples were examined using this approach, and the results agreed with those obtained from more extensive separation followed by high-performance liquid chromatography. Because the threshold can be set at the 3 sigma limit, reliable screening can be accomplished with an error rate of less than 0.26%. Based on this investigation, a high-throughput screening method for ENRO in chicken tissue is proposed.     

Rapid fluorescence screening assay for enrofloxacin and tetracyclines in chicken muscle

A simple, rapid fluorescence assay was developed for screening both enrofloxacin (ENRO) and tetracyclines in chicken muscle at the U.S. tolerance levels (300 ng/g and 2 microg/g, respectively). Screening for both classes of antibiotics is accomplished using one extraction, thus simplifying and expediting the process. The method requires an initial extraction of chicken muscle with 1% acetic acid in acetonitrile, centrifugation, and analysis of the supernatant for ENRO fluorescence. After addition of ammonium hydroxide, magnesium chloride, and methanol, followed by centrifugation and filtration, the supernatant can be measured for tetracycline fluorescence. Chlortetracycline (CTC) was chosen as a representative tetracycline to demonstrate the method, as it displays intermediate sensitivity among the three tetracyclines approved in the U.S. Comparison of the fluorescence of control and tolerance-level-fortified samples of both ENRO and CTC shows no overlap. Setting a threshold as the average fortified fluorescence minus 3sigma allows for successful screening, as illustrated with blind samples as controls or fortified with ENRO and/or CTC over a range of concentrations. This method can provide an alternative or supplemental approach to currently used microbial screening assays.     

Group G streptococcal toxic shock-like syndrome in three cats

Three 8-week-old kittens were presented with a history of acute, generalized weakness and severe fever. One cat was dead upon presentation, and necropsy findings were supportive of a group G Streptococcus spp. septicemia. During their clinical courses, two of the three kittens developed a progressive, marked swelling of one or more limbs. One moribund and severely hypothermic cat was euthanized a few hours after presentation, and necropsy was also supportive of a group G Streptococcus spp. septicemia. One kitten recovered. Group G streptococcal toxic shock-like syndrome was suspected because of the fulminant progression of the septicemia.     

Color stability, reducing activity, and cytochrome c oxidase activity of five bovine muscles

The objective was to characterize the beef psoas major (PM), longissimus lumborum (LL), superficial semimembranosus (SSM), deep semimembranosus (DSM), and semitendinosus (ST) muscles for differences in instrumental and visual color, metmyoglobin-reducing activity (MRA), total reducing activity (TRA), and cytochrome c oxidase activity. The LL and ST had the most color stability and MRA (p < 0.05), the DSM and PM had the least (p < 0.05), and values for the SSM were intermediate. Visual color (r = -0.66) and a and chroma (r = 0.68) were more correlated with MRA than with TRA (r < 0.14 for all measures). This research supports previous reports that color stability among muscles is variable and that MRA is more useful than TRA for explaining the role of reducing activity in muscle-color stability.