Rapid Identification of Mycoplasma pulmonis Isolated from Laboratory Mice and Rats Using Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry

Mycoplasma species identification is based on biochemical, immunological, and molecular methods that require several days for accurate identification. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a novel method for identification of bacteria and has recently been introduced into the clinical microbiology laboratory as a rapid and accurate technique. This method allows a characteristic mass spectral fingerprint to be obtained from whole inactivated mycoplasmal cells. In this study, we evaluated the performance of the MALDI-TOF MS for the identification of Mycoplasma by comparison with standard sequence analysis of 16S rRNA. We developed the first database of MALDI-TOF MS profiles of Mycoplasma species, containing Mycoplasma pulmonis, M. arthritidis, and M. neurolyticum, which are the most common pathogens in mice and/or rats, and species-specific spectra were recorded. Using the database, 6 clinical isolates were identified. Six tracheal swabs from 4 mice and 2 rats were cultured on PPLO agar for 4 to 7 days, and the colonies were directly applied to analyze the protein profiles. Five strains were identified as M. pulmonis, and 1 strain from a mouse was identified as M. neurolyticum (spectral scores were >2.00); the results were consistent with the results of the 16S rRNA gene sequence analysis (homologies>97.0%). These data indicate that MALDI-TOF MS can be used as a clearly rapid, accurate, and cost-effective method for the identification of M. pulmonis isolates, and this system may represent a serious alternative for clinical laboratories to identify Mycoplasma species.     

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Ameliorative effect of K-type and Ca-type artificial zeolites on the growth of beets in saline and sodic soils

Abstract

Beets were grown on soils with various exchangeable sodium percentages (ESP). A saline non-sodic soil (SA, ESP = 3.2), a saline sodic soil (SO, ESP = 23), and a saline high sodic soil (HSO, ESP = 78) were prepared from Tottori sand dune soil (CO). K-type and Ca-type artificial zeolites (50 g kg^?1) were applied to these soils in order to evaluate their effects on the chemical properties of saturation extracts of the soils, water deficit, cation uptake and transport, and cation balance of beet plants. In the zeolite-free treatments, beet growth was accelerated in SA and SO, but was suppressed in HSO compared with CO. The addition of both types of zeolites ameliorated plant growth in all the soils studied, especially HSO. The relative dry weight of the soils treated by the K-type zeolite to the zeolitefree soil was 189% for CO, 125% for SA, 130% for SO, and 222% for HSO. For the soils treated with the Ca-type zeolite, the values were 169, 116, 132, and 341%, respectively. In SA, SO, and HSO, the addition of the K-type zeolite increased the K uptake due to the increase of the K concentration of saturation extracts of soils. The addition of the Ca-type zeolite increased the Ca uptake due to the increase in the Ca concentration of the saturation extracts of soils which was accompanied by an increase in the K uptake. The increase in the uptake of K or Ca and decreased in the transport of Na by the addition of both types of zeolites improved the cation balance of the plants. The Ca-type zeolite did not increase the water deficit even though it increased the electric conductivity in all the soils. The results indicated that both types of artificial zeolites were able to improve the growth performance of beets in saline and sodic soils and that the K-type zeolite could be used as a K-fertilizer as well.     

In vitro differentiation of canine celiac adipose tissue-derived stromal cells into neuronal cells

To investigate in vitro differentiation of canine adipose tissue-derived stromal cells (ATSCs) into neuronal cells, ATSCs from celiac adipose tissue in clinically healthy beagle dogs were treated with 100 muM dibutyryl cyclic adenosine monophosphate (dbcAMP) and 125 muM isobuthylmethylxanthine (IBMX). ATSCs were morphologically changed into differentiated ATSCs from spindle-shaped cells to neuron-like cells with numerous processes after the treatment. Expression of neuron-specific enolase (NSE) as an early neuron specific marker protein was detected in both ATSCs and differentiated ATSCs, however diachronic increase of NSE expression was observed in differentiated ATSCs after the treatment with dbcAMP/IBMX. In addition, neurofilament-68 (NF-68) as an early to mature neuron specific marker protein was weakly expressed in differentiated ATSCs. Neuron specific glutamate and glucose transporter (EAAC1 and GLUT-3, respectively) mRNAs were strongly expressed in differentiated ATSCs compared with those in ATSCs, although glia specific glutamate transporter mRNA (GLT-1) was also detected in differentiated ATSCs. ATSCs can differentiate into early to mature neuronal cells and are candidate cells for autologous nerve regeneration therapy, although additional research is needed to examine functional characteristics of differentiated ATSCs.