Carbon Sources Assimilation Tests: Comparison Between a Conventional Method and a Microtechnic (API), in Study of Enterobacteriaceae

Steigerwalt

Boemare³

et al. 1984

A collection of 30 clones derived from 20 luminous and nonluminous Xenorhabdus strains was studied by using deoxyribonucleic acid (DNA) relatedness and electron microscopy, as well as growth factor requirement, nutritional ability, and other biochemical tests. Nonluminous Xenorhabdus cells each contained a crystal-like structure, whereas the cells of luminous strains each contained an accumulation of amorphous material associated with mesosomes and microtubules of unknown function. Two DNA relatedness groups of nonluminous strains were delineated, one of which corresponded to Xenorhabdus nematophilus, a species associated with the nematode Steinernema feltiae (= Neoplectana carpocapsae). The other group contained yellow-pigmented bacteria associated with Steinernema bibionis and new Steinernema species. No definite phenotypic characteristic (except pigmentation) separated the two DNA relatedness groups of nonluminous Xenorhabdus strains. Four DNA relatedness groups were delineated among the luminous clones studied. These four groups could be separated by biochemical tests. Unfortunately, different clones derived from the same strain belonged to different DNA relatedness groups. This happened with the type strain of Xenorhabdus luminescens.Among the nematodes belonging to the order Rhabditida, two families, the Steinernematidae and the Heterorhabditidae, are composed of organisms whose larvae can penetrate living insects and kill them while developing in their bodies (24). These nematodes contain bacteria in their digestive tracts. The nematode genus Steinernema (now including Neopfectana [29]) is associated with the bacterial species Xenorhabdus nematophilus, whereas the nematode genus Heterorhabditis is associated with the bacterial species Xenorhabdus luminescens (19,23,25,28). Although other bacteria are associated with Steinernema spp. (22; N. Boemare, Ph.D. thesis, UniversitC des Sciences, Montpellier, France, 1983), the presence of Xenorhabdus spp. is essential for the mass production of the nematodes used in insect pest control projects ( 5 ; Boemare, Ph.D. thesis).The genus Xenorhabdus Thomas and Poinar 1979 has been assigned to the family Enterobacteriaceae (28).The purposes of this work were to (i) characterize the species of Xenorhabdus by electron microscopy, (ii) characterize the species of Xenorhabdus by using tests commonly used to describe species in the Enterobacteriaceae, (iii) identify growth factor requirements and determine the nutritional abilities of Xenorhabdus strains, and (iv) delineate the species of Xenorhabdus by deoxyribonucleic acid (DNA)-DNA hybridization. MATERIALS AND METHODSBacterial strains. All strains were purified by picking a single colony on a nutrient agar (Difco Laboratories, Detroit, Mich.) plate. When more than one colonial form were produced by a strain, different colonies were picked and grown separately as different clones. The Xenorhabdus * Corresponding author.strains and clones used in this study are listed in Morphological characterization. Two-day-old culture...

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Section: Nematophilvsmentioning

confidence: 99%

Deoxyribonucleic Acid Relatedness and Phenotypic Study of the Genus Xenorhabdus

Steigerwalt

Boemare³

et al. 1984

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“…The defined medium used with these galleries was provided by API System. These galleries have been described and evaluated previously (11). In method €3 we used filter-sterilized M70 minimal medium (28) without agar.…”

mentioning

confidence: 99%

Taxonomy of the Genus Acinetobacter with the Recognition of Acinetobacter baumannii sp. nov., Acinetobacter haemolyticus sp. nov., Acinetobacter johnsonii sp. nov., and Acinetobacter junii sp. nov. and Emended Descriptions of Acinetobacter calcoaceticus and Acinetobacter lwoffii

Bouvet

1986

571

349

“…Attempts to determine growth factor requirements were done by using M70 minimal medium without agar (21) 20186HG01' 21297HG02 21800HG03 21801HG04 21845HG05 2 1927HG06 21929 21930HG07 21998HGO8 21999HG09 22176HG10 22414HGll 22415HG12 22416HG13 225298614 22530HG15 23012HG17 23013HG 18 23027HG19 23149HG20 23166HG21 23167HG22 23168HG23 23169HG24 23170HG25 23227HG26 23236HG27 23237HG28 23269 23270 23271 23272 23273 23274 23275 23302 23303 23367 23436 23471 23506 23516 23640 23641 23674 23677 23678 23681 23706 23777 23795 23879 23878 3-82 5-82 6-82 7-82 8-82 9-82 5-81 Pool Assimilation tests. Carbon source utilization tests were done at the IP by using specially manufactured API strips (API System, La Balme les Grottes, France) which contained pure carbon sources and were similar to commercial API CH, API AO, and API AA galleries (9), except that the total number of tests were limited to 99 carbon sources (these compounds are listed below). The minimal medium without growth factors (supplied by API System) was supplemented with the following 31 growth factors (final concentration in micrograms per milliliter): biotin and thiamine, 0.002; para-aminobenzoic acid, folic acid, nicotinamide, nicotinic acid, pantothenic acid, and pyridoxine, 0.08; riboflavin, 0.4; inositol, 0.8; sodium thiosulfate, 8; and L-alanine, L-arginine, L-aspartic acid, L-cysteine, L-glutamic acid, Lglutamine, glycine, histidine, isoleucine, L-leucine, L-lysine, methionine, ornithine, L-phenylalanine, proline, serine, threonine, L-tryptophan, L-tyrosine, and L-valine, 17.…”

mentioning

confidence: 99%

Budvicia aquatica gen. nov., sp. nov.: a Hydrogen Sulfide-Producing Member of the Enterobacteriaceae

Bouvet

Richard

et al. 1985