Prevalence and diversity of Wolbachia bacteria infecting insect pests of stored products

Wolbachia are common bacterial symbionts of insects. Because infections of these bacteria can alter the reproduction and biology of the host, there is interest in the potential use of Wolbachia to control populations of pest species. To advance this research, we screened 38 species (88 populations from 16 countries) of stored-product insect pests for Wolbachia infections. Infections were detected in nine species of Coleoptera (Anobiidae – Lasioderma serricorne (Fabricius), Stegobium paniceum (Linnaeus); Curculionidae – Sitophilus oryzae (Motschulsky), Sitophilus zeamais Motschulsky; Dermestidae – Attagenus unicolor (Brahm), Dermestes lardarius Linnaeus; Silvanidae – Oryzaephilus mercator (Fauvel), Oryzaephilus surinamensis (Linnaeus); Tenebrionidae – Tribolium confusum Jaquelin Du Val), one species of Hymenoptera (Trichogrammatidae – Trichogramma deion Pinto & Oatman), and one species of Lepidoptera (Pyralidae – Ephestia kuehniella Zeller). Phylogenetic analyses based on wsp gene sequences identified all Wolbachia isolates as members of Supergroups A or B. Additional sequences for the genes gatB, coxA, hcpA, fbpA and ftsZ were obtained for use in multilocus sequence typing (MLST) to identify the sequence types to which the isolates belonged. Our results identify five new Wolbachia-insect associations and support previous findings that infections of Wolbachia in stored-product insect pests are members of Supergroups A or B.     

Screening stored wheat beetles for reproductive parasitic endosymbionts in central Turkey

Stored-product pest insects cause significant loss in stored wheat worldwide. In Turkey, an important wheat producer and historic centre of wheat domestication, almost 60 stored-product pest insects have been reported so far, most of them being coleopteran species. Using reproductive parasitic endosymbionts (RPEs) is a promising recent approach among control methods alternative to insecticides. For planning and studying pest management with these bacteria, first of all, their presence in the natural pest populations should be investigated. The present study focused on screening the RPEs in Central Anatolian stored wheat pests. We collected pests in granaries in Kırşehir province and identified 10 coleopteran species both morphologically and genetically; namely, Ahasverus advena, Cryptolestes ferrugineus, C. pusillus, Carpophilus obsoletus, Oryzaephilus surinamensis, Rhyzopertha dominica, Sitophilus granarius, S. oryzae, S. zeamais, and Tribolium castaneum. In these pests, we screened the most commonly studied RPEs worldwide, Arsenophonus, Cardinium, Fritschea, Hamiltonella, Rickettsia, Spiroplasma, and Wolbachia, by using specific genetic primers. As a result, we detected RPE presence in almost all sampling localities visited. The RPEs that we found were Rickettsia, Spiroplasma, and Wolbachia. We found no infection caused by Arsenophonus, Cardinium, Fritschea, nor Hamiltonella. Rickettsia presence was only in S. granarius populations, whereas Spiroplasma and Wolbachia presence were not species specific. 22% of all sampled beetle individuals were Wolbachia positive. The highest detection rate per granary was that of Spiroplasma (80%). Wolbachia and Spiroplasma were the most frequently detected RPEs per insect species. We also found several cases of coinfections. This study is the first attempt to screen stored-product pests for seven RPEs together.     

The efficacy and persistence of diatomaceous earths admixed with commodity against four tropical stored product beetle pests

The efficacy and persistence of two commercially available enhanced diatomaceous earth (DE) products (Dryacide^® and Protect-It^®) against four common tropical storage pests (Prostephanus truncatus, Sitophilus zeamais, Callosobruchus maculatus and Acanthoscelides obtectus) were studied when admixed with typical host commodities at different application rates and relative humidities. Persistence of the enhanced DE treatments was considered after 3 and 6 months storage by assessment of both adult mortality and F1 progeny emergence. Both DEs usually increased parental mortality and reduced progeny emergence of all four insect species in comparison with the untreated control at both 50% and 60% r.h., and at all storage periods. However, efficacy was inversely related to duration of storage and over time the host commodity also became less suitable for insect development. Each insect species differed in its susceptibility to the DE treatments, highlighting the need for field application rates to be based upon the entire spectrum of pest species likely to be present during storage.     

Factors affecting the behaviour of beetle pests in stored grain, with particular reference to the development of lures

This review summarises the published information on the behaviour of beetles in grain bulks that could lead to the identification of additional factors with the potential for enhancing existing lures or developing new ones. The review concentrates mainly on the adult stage of the five beetle pest species Oryzaephilus surinamensis (L.), Sitophilus granarius (L.), Cryptolestes ferrugineus (Stephens), Typhaea stercorea (L.) and Ahasverus advena (Waltl). After an analysis of the grain store environment and the origins of storage pests, particular attention is paid to behaviour associated with beetle movement and distribution within grain bulks, and their response to food and other multi-species attractants. Potential sources of new semiochemicals are identified; as well as the grains themselves, these include attractive volatiles associated with hot spots in grain, mouldy grain, and alternative insect pest habitats such as mammals’ and birds’ nests. It might also be worth looking for new semiochemicals produced during the mass upward movements of beetles from grain often seen following disturbance, although these could be repellents rather than attractants. Other sources of repellent semiochemicals with potential as grain protectants could be predatory and parasitic species of insects. Consideration is given to behavioural influences that may conflict with the intended response to attractive lures; for example, the presence of damaged grains, moisture and temperature gradients in the grain bulk may act as rival attractants, while many species prefer to hide away from bright light. As population densities rise, inter- and intra-specific competition becomes increasingly important. Human activities associated with grain handling, drying, cooling and pest control, as well as the presence of natural enemies and pathogens, all have the potential to affect insect response to a lure. Opportunities for the development of more effective lures are highlighted.     

Is the production of the biogenic amines tyramine and putrescine a species-level trait in enterococci?

Biogenic amines (BA) are toxic nitrogenous compounds that can be accumulated in foods via the microbial decarboxylation of certain amino acids. Lactic acid bacteria (LAB) strains belonging to different species and genera have been described as BA producers and are mainly responsible for their synthesis in fermented foods. It is generally accepted that the capacity to produced BAs is strain-dependent. However, the large number of enterococci identified as BA producers suggests that the aminogenic trait may be a species-level characteristic. Enterococcus faecalis, Enterococcus faecium and Enterococcus durans strains of different origin were analysed to determine their capacity to produce tyramine and putrescine. The presence of the genes responsible for this and the identity of their flanking regions were checked by PCR. The results suggest that tyramine biosynthesis is a species-level characteristic in E. faecalis, E. faecium and E. durans. Putrescine synthesis was found to be a species-level trait of E. faecalis, with production occurring via the agmatine deamination pathway. Some E. faecium strains of human origin also produced putrescine; this trait was probably acquired via horizontal gene transfer.     

Effect of low pressures on the survival of cocoa pests at 18°C

This study forms part of an effort to eliminate the need for fumigation with methyl bromide to control insect infestations in stored cocoa beans, through development of novel alternative vacuum-hermetic technology. In this communication, the effects of low pressures and exposure time were studied on the mortality of insects at a temperature of 18°C, chosen to simulate cocoa bean storage conditions in temperate climates.Three insect species were used, two of which are major pests of cocoa beans in producer countries, Ephestia cautella (Walker), and Tribolium castaneum (Herbst), while the third, Oryzaephilus surinamensis (L.), is a potential storage pest in temperate climates. For T. castaneum and E. cautella the egg stage was the most resistant to 55±10 mm Hg at 18°C, the times needed to obtain 99% egg mortality were 96 and 149 h, respectively. For O. surinamensis, the adult stage was the most resistant with 164 h being required to obtain 99% mortality.     

Insecticidal effect of Steinernema feltiae (Filipjev) (Nematoda: Steinernematidae) against Tribolium confusum du Val (Coleoptera: Tenebrionidae) and Ephestia kuehniella (Zeller) (Lepidoptera: Pyralidae) in stored wheat

The confused flour beetle, Tribolium confusum, and the Mediterranean flour moth, Ephestia kuehniella, are important pests of stored grain products. The insecticidal effect of three strains (UK 76 [=Nemasys], USA/SC, and Hawaii) of the entomopathogenic nematode Steinernema feltiae was determined in the laboratory, in wheat, against these pests. The nematodes were applied at three dose rates: 100, 300 and 900 nematodes/insect individual. The Hawaii strain was most virulent against T. confusum adults and larvae, with a significant dose effect in the case of the larvae. Larval mortality of this species reached 79% and 100% after 7 and 14 d of exposure to the nematodes, respectively, at the highest dose applied. On the other hand, adult mortality of T. confusum did not exceed 66%. In the case of E. kuehniella larvae, USA/SC performed best causing 52% and 69% mortality after 7 and 14 d exposure, respectively, at the highest dose tested. Since very few data are available on the effect of entomopathogenic nematodes against these pests, it is concluded that the Hawaii and USA/SC strains of S. feltiae should be further investigated as promising biological control agents for T. confusum and E. kuehniella.     

Ecological factors associated with pest status in Callosobruchus (Coleoptera: Bruchidae): high host specificity of non-pests to Cajaninae (Fabaceae)

Larval host plants of six Callosobruchus species (Coleoptera: Bruchidae), including pest species, were discovered by collecting the seeds of wild (or inedible) and cultivated edible legumes in the field in Taiwan, Thailand, continental China, Myanmar, Nepal and the Philippines. A close relationship between Asian Callosobruchus species and the leguminous subtribe Cajaninae was revealed: Rhynchosia species were commonly used by three Callosobruchus species, wild and cultivated species of Cajanus and two Dunbaria species, respectively, by single species. Two Taiwanese species were confirmed to be conspecific with continental species. We further reviewed host legumes of 11 species of Callosobruchus including nine species of pests. There were significant positive correlations between geographic distribution range and host range at all three taxonomic levels of hosts. Principal component analysis on geographic distribution range, host range, altitude of distribution, utilizations of cultivated hosts, of Cajaninae and of Phaseolinae showed that the first axis (PC1) described 52% of total variance, which was related significantly with the frequencies of utilization of cultivated legumes (0.93), and of Cajaninae (Cajanus, Dunbaria and Rhynchosia) (−0.85). PC1 was also positively correlated with the frequency of utilization of Phaseolinae (Vigna and Lablab) (0.68), geographical range (0.67) and with host range (0.67) before Bonferroni corrections. Contrary to the polyphagy of the widely distributed pest Callosobruchus, non-pest species exhibited fidelity to single specific genera of wild or inedible legumes, and pests with limited distribution are specific to leguminous subtribes. Non-pests are characterized by tight association with Cajaninae. We concluded that specialization to non-economic Cajaninae prevents a species of Callosobruchus from becoming a pest of cultivated legumes.     

Analysis of free fatty acids in food substrates and in the dust and frass of stored-product pests: Potential for species discrimination?

The larger grain borer, Prostephanus truncatus, is a serious beetle pest that tunnels extensively to produce large quantities of dust and frass. The natural enemy Teretrius nigrescens is an important biological control beetle which is known to exploit at close-range solvent-extractable chemical cues in the dust and frass. The objective of the current study was to analyse quantitatively and qualitatively, the free fatty acid mixtures in different food-substrate materials both before and after insect attack by a range of stored-product pests in order to ascertain whether differences in these mixtures could explain the T. nigrescens selectivity to P. truncatus dust/frass over that of other species irrespective of food substrate. By TLC, GC and GC-MS we found triglyceride and five free fatty acids were the most abundant chemicals in dust/frass (palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1), linoleic acid (C18:2) and linolenic acid (C18:3)). In maize flour, Sitophilus species did not significantly change free fatty acid concentrations whereas with P. truncatus, Rhyzopertha dominica and Dinoderus minutus there were 4-6-fold increases, and, for Tribolium species there were over 20-fold increases. These differences provide interesting insights to tunnelling/feeding habits and are correlated with known feeding preferences within grain. Principal component analysis (PCA) demonstrated that free fatty acid ratios in dust/frass of different species are most linked to the food substrate and confer little discriminatory information that could be used to distinguish between the different species. Although increases in free fatty acid concentrations are good indicators of pest infestation and this may contribute behaviourally in an additive or synergistic way, we conclude that other chemical(s) are present and are key to T. nigrescens recognition of P. truncatus on different substrates.     

Fecundity and ability of the parasitoid Lariophagus distinguendus (Hymenoptera: Pteromalidae) to find larvae of the granary weevil Sitophilus granarius (Coleoptera: Curculionidae) in bulk grain

Fecundity and host finding behavior in the storage environment were examined for the parasitoid Lariophagus distinguendus (Förster), a potential agent for biological control of stored-product pest beetles. Larvae of the granary weevil, Sitophilus granarius (L.), in wheat were used as hosts. The fecundity varied considerably among strains of different geographical origin, from 1.3±2.4 to 92.5±23.6 offspring per female parasitoid. Host finding ability was examined in a small acrylic cylinder (about 42 kg wheat), a silo-bin (20.5 t wheat grain), and the flat storage grain bin in a commercial facility (3000 t rye). Experiments in the cylinder and the silo-bin revealed that the number of hosts found by the parasitoids decreases with increasing depth. Moreover, parasitoids were more likely to find hosts in the cylinder than under field storage conditions in a silo-bin. This indicates that the density of parasitoids per unit volume of the grain might be an important factor in determining how deep they penetrate a grain layer. In the silo-bin and the commercial grain store, parasitoids were able to find and parasitize hosts located up to 4 m vertically and horizontally from the release point. The significance of these results with respect to the suitability of L. distinguendus for biological control of the granary weevil is discussed.     

Discrimination of dairy industry isolates of the Lactobacillus casei group

Lactobacilli are a major part of the microflora of the gut and of many fermented dairy products, and are found in a variety of environments. Lactobacillus casei, Lactobacillus paracasei, Lactobacillus rhamnosus, and Lactobacillus zeae form a closely related taxonomic group within the facultatively heterofermentative lactobacilli. The classification and nomenclature of these bacteria are controversial. In this study, relationships between these species were investigated using type strains and dairy industry isolates examined with DNA-based techniques and conventional carbohydrate use tests. Carbohydrate use patterns gave poor discrimination of some species, but DNA PCR using specific primers targeted to sequences of the 16S rRNA gene discriminated 4 types consistent with the currently recognized species. Pulsed-field agarose gel electrophoresis of chromosomal NotI restriction fragments identified 18 different band patterns from 21 independent Lactobacillus isolates and confirmed the identity of L. casei strains from 2 culture collections (CSCC 5203 and ASCC 290), both representing the type strain of L. casei. Some isolates were reclassified as L. rhamnosus, suggesting that the prevalence of L. rhamnosus as a natural component of the microflora of dairy foods and dairy environments has previously been underestimated. These methods can provide a practical basis for discrimination of the species and identification of individual industrial strains.     

Resistance of Tripsacorn to Sitophilus zeamais and Oryzaephilus surinamensis

One strategy that has been used to find germplasm for developing improved plant varieties is to test ancestral germplasm for the desired traits. Although the progenitors of commercial maize are not known, a hybrid (called Tripsacorn) developed from a perennial teosinte, Zea diploperennis, and eastern gamagrass, Tripsacum dactyloides, resembles the earliest known samples of primitive domesticated maize. We tested resistance of whole Tripsacorn to the primary storage pest (primary storage pests can infest intact kernels) the maize weevil, Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), and resistance of ground Tripsacorn to the secondary storage pest (secondary pests usually cannot infest intact kernels) the sawtoothed grain beetle, Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae). Tripsacorn was immune to attack by S. zeamais. The weevils were unable to lay eggs in the Tripsacorn, and we hypothesized that the hardness of the fruitcase was responsible for lack of weevil oviposition. Oryzaephilus surinamensis were able to complete immature development on ground Tripsacorn, but duration of development was longer and weight of emerged adults was less than for beetles developing on wheat. Hardness of the fruitcase may have been a primitive mechanism of defense against insects and other pests, but probably would not be an acceptable trait in commercial varieties. It remains to be determined whether the possible antibiotic effect demonstrated in ground Tripsacorn would be a useful trait in commercial maize hybrids.     

Use of recA gene sequence analysis for the identification of Staphylococcus equorum strains predominant on dry-cured hams

Spanish dry-cured ham is an uncooked meat product highly appreciated due to its characteristics flavour. In this study, we examined the accuracy of biochemical tests and 16S rDNA sequencing in the identification of 56 staphylococcal strains isolated during industrial Spanish dry-cured ham processes. Important differences were observed comparing genotypic and phenotypic data. Staphylococcus xylosus was the prevalent species identified by biochemical methods (87.5%), however, sequencing of the 16S rDNA resulted in an unambiguous identification of Staphylococcus equorum (73.2%) and Staphylococcus vitulinus (8.9%) strains. Reliable identification of meat staphylococci, mainly among S. xylosus and S. equorum strains could be also achieved by means of recA gene sequence comparison. Two degenerate primers previously described for lactic acid bacteria were used to amplify an internal fragment of the recA gene. This fragment was amplified from twelve staphylococcal type strains representing frequent meat species. The results indicated that recA sequencing is an adequate method to discriminate among meat staphylococci. In addition, S. xylosus and S. equorum strains could be more accurately discriminated by recA sequencing than 16S rDNA or sodA sequencing. The S. equorum sequence diversity showed at the intra-species level by recA gene sequencing confirmed the high heterogeneity described among S. equorum strains.     

Susceptibility of commercial oat cultivars to Cryptolestes pusillus and Oryzaephilus surinamensis

Susceptibility to two storage insect pests [(Cryptolestes pusillus (Schönherr) and Oryzaephilus surinamensis (L.)] of eight commercial oat cultivars from the United States was determined in laboratory studies. Duration of insect development was shorter and number of progeny produced was greater on cracked than on whole oats. Simulations based on data from the study showed that insect populations would reach the threshold level for treatment in 2-3 months of storage at 30°C on cracked oats. Insect population development was slowest on the hulless cultivar Paul when the oat kernels were cracked. Simulations also indicated that all cultivars of whole oats tested could be stored for at least 1 yr at 30°C without reaching the threshold for treatment when infested with these two species of insects, and insect populations would decrease over time on the cultivars Don, Jerry, Milton, NewDak, Otana, and Valley. Analyses of oat grain quality characteristics, including kernel weight, groat hardness, and groat composition, provided little insight into the mechanism of observed differences in insect development among cultivars. Hardness of the kernels (as indicated by % broken groats after dehulling) may be related to near immunity to these two species of insects in whole Otana. Steaming whole oats to inactivate hydrolytic enzymes in the trichomes of the pericarp did not increase susceptibility to these two species of insects, suggesting that enzymes in the trichomes were not responsible for insect population development being slower on whole oats than on cracked oats. Although we were unable to identify the factors that determined relative susceptibility in this study, the results will be useful for selecting commercial oat cultivars for planting that will be less susceptible to insect pests in storage and suggest that the economics of cleaning oats before storage to reduce insect population growth should be investigated.     

Genome size and base composition in Oryzaephilus surinamensis (Coleoptera: Sylvanidae) and differences between native (feral) and silo pest populations in Israel

In this study, flow cytometry was used for assessing and comparing the genome size (GS) and the whole genome base composition (AT/GC ratio) of the saw-toothed grain beetle Oryzaephilus surinamensis (L.). In addition, the presence and frequency of endosymbiotic Wolbachia bacteria was studied. The haploid GS was estimated to lie within the range of 151.5-154 Mbp in O. surinamensis, making it the smallest value of haploid GS known among beetles. Furthermore, it was found that in eight silo pest populations GS was significantly smaller than in eight feral (native) populations obtained from fallen oak acorns. The ability of O. surinamensis to colonize different habitats globally could be connected with an unusually AT-rich (for an invertebrate) genome (AT-base content ranging from 68 to 76%). Native (feral) populations of O. surinamensis appear to have genetically diverged from the storage-pest populations tested. Larvae of pest origin survived better than larvae of native (feral) origin under laboratory conditions, which resembled silo conditions more than natural habitats.     

Wolbachia induces costs to life-history and reproductive traits in the moth,Ephestia kuehniella

The intracellular endosymbiont Wolbachia pipientisis is well-known as one of the most common bacterial symbionts of arthropods. Recently, research has focused on the potential to utilize Wolbachia as a biocontrol agent of agricultural and medical pest insect species. Wolbachia blocks host infection from other pathogens and viruses in some species, however, it can also influence host life-history and reproductive traits. Therefore, in order to understand the biological impact and potential economic utility of Wolbachia, it is necessary to investigate the effects of Wolbachia infection on host traits. We compared life-history and reproductive traits between Wolbachia-infected and cured population in Mediterranean Flour Moth, Ephestia kuehniella. E. kuehniella is well known as a pest of stored products, and when infected with Wolbachia, it exhibits cytoplasmic incompatibility between uninfected females and infected males. We found that E. kuehniella suffers costs as a result of Wolbachia infection, through decreased larval survival and adult longevity, and prolonged developmental period. Moreover, reproductive performance was greater in the uninfected population, when excluding the effect of cytoplasmic incompatibility. Our results indicate that E. kuehniella suffers deleterious effects on both life-history and reproductive traits as a result of being infected with Wolbachia. We suggest such costs should be considered when evaluating the efficacy of utilizing Wolbachia in pest control.     

Biocontrol as a strategy to reduce the impact of ochratoxin A and Aspergillus section Nigri in grapes

The efficacy of two strains of Kluyveromyces thermotolerans in preventing the growth and ochratoxin A (OTA) accumulation of ochratoxigenic fungi both “in vitro” and “in situ” was evaluated. The data from this study showed that both yeast strains were able to control Aspergillus carbonarius and A. niger aggregate species growth and ochratoxin A accumulation. The inhibitory effects were dependent on the ochratoxigenic species, yeast strains, a_w and temperature evaluated and their interactions. Over all conditions assayed, ochratoxin A accumulation was reduced from 3% to 100% and the growth rate from 11% to 82.5%, depending on conditions. These results are promising for future development of a bio-pesticide.     

Distribution of the Medea factor M in populations of Tribolium castaneum (Herbst) in the United States

The distribution of the maternally acting, selfish gene Medea⁴ (M⁴) was determined in populations of the red flour beetle, Tribolium castaneum (Herbst), collected in the southern and midwestern United States. We found clear evidence for the existence of two major regional subpopulations, with a boundary that roughly corresponds to 33°N latitude. All 26 strains collected in 10 states north of this latitude were homozygous for the M⁴ allele, while only two of 29 strains collected in six states south of this latitude were homozygous for the allele. Of the remaining 27 southern strains, 21 lacked the M⁴ allele entirely, while six contained a mixture of M⁴ and non-M⁴ alleles. This is the first evidence of either the existence of biotypes or the presence of major barriers to gene flow in wild populations of this ubiquitous insect species.     

Phylogenetic analyses and toxigenic profiles of Fusarium equiseti and Fusarium acuminatum isolated from cereals from Southern Europe

Fusarium equiseti and Fusarium acuminatum are toxigenic species that contaminate cereal crops from diverse climatic regions. They are common in Spanish cereals. The information available on their phylogenetics and toxigenic profiles is, however, insufficient to assist risk evaluation. In this work, phylogenetic analyses were performed using partial sequences of the translation elongation factor gene (EF-1α) of F. equiseti and F. acuminatum strains isolated from barley and wheat from Spain and other countries. The Northern and Southern European F. equiseti strains largely separated into two phylogenetically distinct clusters. This suggests the existence of two distinct populations within this species, explaining its presence in these regions of markedly different climate. Production of type A and B trichothecenes by the Spanish strains, examined in wheat cultures using a multitoxin analytical method, indicated that F. equiseti could produce deoxynivalenol and nivalenol and other trichothecenes, at concentrations that might represent a significant risk of toxin contamination for Southern European cereals. F. acuminatum showed low intraspecific genetic variability and 58% of the strains could produce deoxynivalenol at low level. Neither species was found to produce T-2 or HT-2 toxins. The present results provide important phylogenetic and toxigenic information essential for the accurate prediction of toxigenic risk.     

Diversity of stress tolerance in Lactobacillus plantarum, Lactobacillus pentosus and Lactobacillus paraplantarum: A multivariate screening study

Sixty-three strains of the taxonomically related species Lactobacillus plantarum subsp. plantarum, L. plantarum subsp. argentoratensis, L. paraplantarum and L. pentosus isolated from sourdoughs and other food and non-food sources and 14 strains of other members of the genus Lactobacillus were screened for their tolerance of acid, alkaline, heat, oxidative, osmotic, detergent and starvation stresses in order to evaluate the diversity of stress response. Most strains of the L. plantarum group were highly tolerant of acid, alkaline and osmotic stress and highly sensitive to detergent stress, while a larger diversity was found for other stress. Multivariate analysis allowed grouping the strains in clusters with similar response patterns. Stress response patterns in the L. plantarum group were similar to those of species of the L. casei/L. paracasei group but clearly different from those of other mesophilic Lactobacillus. No relationship was found between grouping obtained on the basis of stress response patterns and by genotypic fingerprinting (rep-PCR), nor with the taxonomic position or isolation source of the strains. Further experiments with selected strains showed that exponential phase cells were generally but not always more sensitive than stationary phase cells. The ability to grow under stressful conditions showed a slightly better correlation with the ecological conditions prevailing in the isolation niches of the strains.This study will be the basis for further investigations to identify and exploit the basis of diversity in the stress response of lactic acid bacteria.