Hematoma subcapsular esplénico espontáneo y mononucleosis infecciosa: una asociación de interés para el cirujano

La rotura espontánea de bazo es una complicación infrecuente pero muy grave de la mononucleosis infecciosa. Dada la urgencia con la que suele presentarse tal situación, la esplenectomía suele ser la opción quirúrgica más utilizada. Presentamos el caso de un paciente en el que con el diagnóstico de rotura esplénica espontánea se somete a laparoscopia diagnóstica en la que se consigue la preservación del bazo con buena evolución clínica. A la luz de este paciente, consideramos que, en caso de estabilidad clínica y hematomas subcapsulares de bazo, la laparoscopia con preservación esplénica es una opción viable y con buenos resultados. (provisto por Infomedic International)

Spontaneous rupture of the spleen is a rare but very serious complication of infectious mononucleosis. Given the urgency with which such a situation usually presents, splenectomy is usually the most used surgical option. We present the case of a patient diagnosed with spontaneous splenic rupture who underwent diagnostic laparoscopy in which spleen preservation was achieved with good clinical evolution. In the light of this patient, we consider that, in the case of clinical stability and subcapsular haematomas of the spleen, laparoscopy with splenic preservation is a viable option with good results. (provided by Infomedic International)

Spanish cohort of VEXAS syndrome: clinical manifestations, outcome of treatments and novel evidences about UBA1 mosaicism.

BACKGROUND: The vacuoles, E1-enzyme, X linked, autoinflammatory and somatic (VEXAS) syndrome is an adult-onset autoinflammatory disease (AID) due to postzygotic UBA1 variants. OBJECTIVES: To investigate the presence of VEXAS syndrome among patients with adult-onset undiagnosed AID. Additional studies evaluated the mosaicism distribution and the circulating cytokines. METHODS: Gene analyses were performed by both Sanger and amplicon-based deep sequencing. Patients' data were collected from their medical charts. Cytokines were quantified by Luminex. RESULTS: Genetic analyses of enrolled patients (n=42) identified 30 patients carrying UBA1 pathogenic variants, with frequencies compatible for postzygotic variants. All patients were male individuals who presented with a late-onset disease (mean 67.5 years; median 67.0 years) characterised by cutaneous lesions (90%), fever (66.7%), pulmonary manifestations (66.7%) and arthritis (53.3%). Macrocytic anaemia and increased erythrocyte sedimentation rate and ferritin were the most relevant analytical abnormalities. Glucocorticoids ameliorated the inflammatory manifestations, but most patients became glucocorticoid-dependent. Positive responses were obtained when targeting the haematopoietic component of the disease with either decitabine or allogeneic haematopoietic stem cell transplantation. Additional analyses detected the UBA1 variants in both haematopoietic and non-haematopoietic tissues. Finally, analysis of circulating cytokines did not identify inflammatory mediators of the disease. CONCLUSION: Thirty patients with adult-onset AID were definitively diagnosed with VEXAS syndrome through genetic analyses. Despite minor interindividual differences, their main characteristics were in concordance with previous reports. We detected for the first time the UBA1 mosaicism in non-haematopoietic tissue, which questions the previous concept of myeloid-restricted mosaicism and may have conceptual consequences for the disease mechanisms.

Microbiological and Chemical characterization of water kefir: An innovative source of potential probiotics for bee nutrition / Caracterización microbiológica y química del kéfir de agua: una fuente innovadora de potenciales probióticos para la nutrición de las abejas

Abstract We evaluated the microbial composition of water kefir grains and beverage overthe course of one year to determine whether the number and type of microorganisms changedover the time. Bacteria and yeast colonies with different morphologies were isolated fromwater kefir and their antimicrobial activity was evaluated against Paenibacillus larvae andAscosphaera apis. A chemical characterization of kefir was also carried out. Our results con-firmed that bacteria and yeasts were more numerous in kefir grains compared with those in thebeverage. The counts of microorganisms declined, although an important microbial community was still present in kefir after the long storage period. Eleven strains which inhibited bee pathogens were isolated from kefir. Genotypic results demonstrated that these isolates included Lentilactobacillus hilgardii, Lentilactobacillus buchneri and Saccharomyces cerevisiae. Thus, water kefir may be an innovative source of potential probiotic strains for bee nutrition in order to control honeybee diseases.

Resumen Evaluamos la composición microbiana del kéfir de agua durante un ano para determinar si la cantidad y el tipo de microorganismos cambiaban con el tiempo. Se aislaron colonias de bacterias y de levaduras con diferentes morfologías, y su actividad antimicrobiana se evaluó frente a Paenibacillus larvaey Ascosphaeraapis. También se realizó una caracterización química del kéfir. Nuestros resultados confirmaron que las bacterias y las levaduras eran más numerosas en los gránulos de kéfir en comparación con la parte líquida. Los recuentos de microorganismos disminuyeron, aunque una cantidad igualmente importante se encontró en el kéfir después de un año. Se aislaron del kéfir once cepas que inhibieron los mencionados patógenos de abejas. Los resultados genotípicos demostraron que estos aislamientos eran Lentilactobacillus hilgardii, Lentilactobacillus buchneri y Saccharomyces cerevisiae. Por lo tanto, el kéfir de agua podría ser una fuente innovadora de potenciales cepas probióticas para contribuir a la nutrición y sanidad de las abejas.

Ganoderma sessile is a fast polysaccharide producer among Ganoderma species.

The selection of fast-growing and high-yield-producing strains is required to satisfy the market demand on fungal food supplements. To that aim, three strains deposited in our collection as G. lucidum and G. oregonense were screened for polysaccharide production and biomass yield. Ganoderma strains deposited as G. lucidum were identified as G. sessile and G. lingzhi by nuc rDNA internal transcribed spacer ITS1-5.8S-ITS2 (ITS) and translation elongation factor 1-α (TEF1-α) phylogenies. The identity of G. oregonense was confirmed by molecular phylogeny and biogeography. Additionally, mycelial antagonism confirmed species differentiation, and strains were further distinguished by morphology and protein profiles. Biomass and polysaccharide yields of G. sessile were clearly different from those of G. lingzhi and G. oregonense in both liquid culture and solid-state fermentation. The maximum polysaccharide yield (4.52 ± 0.83 g L-1) for G. sessile was obtained from submerged cultures at day 9. G. sessile also achieved the highest linear growth in lignocellulosic solid substrates. Consequently, basidiomata were successfully obtained by solid-state fermentation in polypropylene bags, whereas G. lingzhi and G. oregonense mushrooms were not produced in artificial solid substrates. G. sessile, a species frequently collected in America, showed to be a promising polysaccharide producer for the manufacture of dietary supplements.

Long-Term Anorectal Manometry Outcomes After Laparoscopic and Transanal Total Mesorectal Excision.

Background: During transanal total mesorectal excision (TaTME), sustained dilation of the anal canal occurs, which can cause anatomical and functional damage to the sphincter complex. This study aimed to analyze the impact of laparoscopic total mesorectal excision (LaTME) and TaTME in anorectal sphincter function. Materials and Methods: An observational study was conducted comparing two cohorts of patients who underwent LaTME or TaTME for rectal cancer. The two groups were paired for comparison based on age, gender, and distance of the neoplasm to the anal margin. The anorectal function was assessed by manometry before surgery and at least 6 months after primary intervention or stoma closure. The intestinal function was assessed using the low anterior resection syndrome (LARS) score. Results: Twenty-two patients were included. There were no significant differences in baseline characteristics between groups except for the time between surgery and testing. A decrease in the mean resting and squeeze pressures between pre- and postoperative manometry was observed in both the treatment groups, the difference being only significant in the squeeze pressure values (TaTME P = .003; LaTME P = .004). After surgery, squeeze pressure reduction correlated with a worsening of the LARS point count (rho 0.587; P = .004). The time elapsed since surgery was negatively correlated with the LARS point count (rho -0.696; P = .001) and the difference between pre- and postoperative mean squeeze pressures (rho -0.499; P = .018). Conclusion: Manometric findings after TME are comparable between the laparoscopic and the transanal approach. Deterioration of both anal sphincter function and LARS improves with time after surgery.

Organización, mantenimiento y preservación de la Colección de Cultivos Bacterianos del Instituto de Ciencias Biológicas de la Universidad de Ciencias y Artes de Chiapas (UNICACH), México / Organization, maintenance, and preservation of the Bacterial Culture Collection of the Biological Sciences Institute, University of Science and Arts of Chiapas (UNICACH), Mexico

Los ceparios o colecciones de microorganismos son fuentes de recursos genéticos cuyo propósito es la preservación de la diversidad biológica, garantizando su disponibilidad para actividades de docencia, investigación y comerciales. En este trabajo se verificó la viabilidad, pureza y características biológicas de las bacterias que conforman el cepario del Instituto de Ciencias Biológicas de la Universidad de Ciencias y Artes de Chiapas, y se organizó y estructuró la información obtenida en un portal virtual, para propiciar la cooperación académica. El cepario cuenta con 33 microorganismos, la mayoría del género Streptococcus y Escherichia (45,1 y 21,2%, respectivamente). Del primer género, se confirmó la identificación de S. pyogenes (40%), exhibiendo la mayoría genes que codifican para DNAsas. Con respecto al segundo género, un 58,3% de las bacterias fueron confirmadas taxonómicamente como E. coli. De esta especie, la colección cuenta con las cepas prototipo causantes de diarrea y que han preservado sus rasgos genéticos por más de cinco años. Dicho acervo ha impulsado actividades de docencia e investigación, a nivel local e internacional. Es importante que los ceparios sean fuentes sustentables de recursos biológicos, para la adquisición y suministro de especies bacterianas, con la finalidad de fomentar la interacción con la comunidad académica.

Strain collections or bacterial culture collections are genetic resources whose purpose is the preservation of biological diversity, ensuring their availability for teaching, research and trade activities. In this work viability, purity and biological characteristics of bacteria from the bacterial collection of the Institute of Biological Sciences, University of Science and Arts of Chiapas were studied. Information was structured and organized in a virtual site, to promote academic cooperation. The strain bank includes 33 microorganisms, most of the genus Streptococcus and Escherichia (45.1 and 21.2%, respectively). For Streptococcus, the identification of S. pyogenes (40%) was confirmed, by determination of most DNAses encoding genes. For Escherichia 58.3% were taxonomically confirmed as E. coli. For this species, the collection includes typical strains that produce diarrhea and their genetic traits have been preserved for more than five years. This bacterial culture collection has stimulated teaching and research activities at local and international levels. Strain collections are important sources of biological material which can provide bacterial species, in order to encourage interaction with the academic community.

Whole exome sequencing for the identification of a new mutation in TGFB2 involved in a familial case of non-syndromic aortic disease.

BACKGROUND: Non-syndromic aortic disease (NSAD) is a frequently asymptomatic but potentially lethal disease characterised by familial cases of thoracic aortic aneurysms and dissections. This monogenic but genetically heterogeneous condition is primarily inherited as an autosomal dominant disorder with low penetrance and variable expression. Mutations in ACTA2, TGFBR1, TGFBR2, MYH11, SMAD3, MYLK, and FBN1 genes have been described but still, there are many unresolved familial cases. METHODS: The whole exome of two distantly related and affected members of a Spanish family with multiple cases of NSAD was analysed through 5500 SOLiD(™) System for the identification of shared and putative pathogenic variants. RESULTS: A new mutation termed c.C1042T:p.R348C (NM_001135599.2) was identified in TGFB2, a gene located in an evolutionary highly conserved region (Chr1: 218,519,577-218,617,961) that has been recently connected to this disease. The analysis of other family members using capillary sequencing confirmed cosegregation of the mutation with the disease and its incomplete penetrance. CONCLUSIONS: The repeated implication of TGFB2 in the development of thoracic aortic aneurysms and dissections suggests that this gene should be considered during genetic diagnosis of this disease. An accurate diagnosis of affected individuals and additional family members at risk allows for a personalised and more efficient gene-based follow-up and treatment. Finally, the reiterative presence of common musculoskeletal and craniofacial additional features in patients with TGFB2 mutations suggests the existence of a new yet undefined connective tissue syndrome responsible for not only aortic dilation, but also for the other extracardiac alterations present in the affected patients.

The use of morphological and histological features as nutritional condition indices of Pagrus pagrus larvae

Morphometrical and histological techniques were employed to characterize Pagrus pagrus larvae nutritional condition. Larvae were reared in laboratory under controlled conditions with the main objective of testing whether these methodologies allowed finding differences between larvae from different feeding treatments. Once yolk was consumed (three days after hatching) larvae were assigned to a feeding treatment: starved during the whole experiment; delayed feeding, starved during three days; fed during the entire experiment. Algae (Nannochloropsis oculata) and rotifers (Brachionus plicatilis) were provided to larvae for feed treatments. Larvae were fixed daily; for morphometrical purposes in 5% formaldehyde solution, and in Bouin for histological sections. Results herein obtained showed that both methodologies are sensitive enough to distinguish larvae characterized by different nutritional condition states obtained from the feeding treatments. Consequently, these methodologies could be employed in wild red porgy larvae in order to asses their nutritional condition. These techniques could also be employed to check larval quality obtained with aquaculture purposes to estimate the effects of changes in rearing protocols or kind of food supply and thus, to guaranty a higher survival of early developmental stages of reared larvae.

Con el objeto de determinar si las técnicas morfométricas e histológicas permiten diferenciar larvas en estado de inanición de las sometidas a tratamientos de alimentación, ejemplares de Pagrus pagrus fueron criados en laboratorio bajo condiciones controladas. Una vez que el vitelo fue consumido (tres días luego de la eclosión) las larvas fueron sometidas a diferentes tratamientos de alimentación: completamente privadas de alimento, privadas de alimento durante tres días y luego alimentadas, y alimentadas durante todo el experimento. Algas (Nannochloropsis oculata) y rotíferos (Brachionus plicatilis) fueron proporcionados a las larvas de los tratamientos de alimentación. Diariamente se fijaron ejemplares de los tres tratamientos en formol 5% para estudios morfométricos y en Bouin para estudios histológicos. Los resultados obtenidos dan evidencias de que las metodologías presentadas son suficientemente sensibles para determinar diferencias en la condición de larvas sometidas a los tratamientos de alimentación. Por lo tanto, podrían ser empleadas para determinar la condición nutricional de las larvas de besugo recolectadas en el mar. Por otra parte, éstas metodologías podrían ser empleadas para determinar la calidad de larvas producidas en acuicultura y evaluar los efectos de modificaciones introducidas en los protocolos de crianza o en el tipo de alimento suministrado con el objeto de mejorar la supervivencia de los estadios tempranos de desarrollo.

Emended descriptions of Bacillus sporothermodurans and Bacillus oleronius with the inclusion of dairy farm isolates of both species.

Bacillus sporothermodurans is an industrially important micro-organism because of its ability to produce endospores which resist ultra-high temperature (UHT) and industrial sterilization processes. It was described by Pettersson et al. (1996) [Pettersson, B., Lembke, F., Hammer, P., Stackebrandt, E. & Priest, F. G. (1996). Int J Syst Bacteriol 46, 759-764] based on seven genetically homogeneous isolates all from UHT milk. Bacillus oleronius, the closest phylogenetic neighbour of B. sporothermodurans, was described by Kuhnigk et al. (1995) [Kuhnigk, T., Borst, E.-M., Breunig, A., König, H., Collins, M. D., Hutson, R. A. & Kämpfer, P. (1995). Can J Microbiol 41, 699-706] based on a single strain, isolated from the hindgut of the termite Reticulitermes santonensis. A polyphasic study of a heterogeneous collection of B. sporothermodurans and B. oleronius strains isolated from various sources and geographical origins led to an emended description of both species. Additional data presented are the results of fatty acid, quinone and/or cell wall (polar lipid) analyses. DNA-DNA hybridization confirmed 3 subgroups of strains obtained after SDS-PAGE analysis of cellular proteins as B. sporothermodurans. One named B. sporothermodurans strain (R-7489) was reclassified as a Bacillus fordii strain. The phenotypic profiles of both species were rather heterogeneous, sometimes different from the original descriptions and did not differ in a large number of characteristics, although B. oleronius generally gave stronger reactions in its positive tests than did B. sporothermodurans; the variable and weak reactions for both organisms with some substrates blurred the distinction between the two. However, differences in polar lipid, SDS-PAGE and menaquinone profiles clearly allow distinction between the two species.

[Pelvic reconstruction after abdominoperineal resection of the rectum]. / Reconstrucción pélvica tras amputación abdominoperineal del recto.

Despite the advances in the treatment of cancer of the rectum and the expansion of the multimodal therapeutic technique, abdominoperineal resection (APR) still needs to be performed as radical treatment in 20-30% of cases. APR of the rectum involves a significant morbidity, including intestinal obstruction and wound complications, with radiotherapy-induced enteritis being able to develop in 15% of cases subjected to post-operative radiotherapy. Furthermore, with the aim of improving local oncology results, an extended APR is recommended; a technique that requires a perineal reconstruction technique that allows a tension free closure in a previously radiated tissue and may prevent perineal hernias developing. The objective of this article is to review pelvic and perineal repair methods after APR due to cancer, with special attention to the new prosthetic repair techniques.

Recent cigarette smoking and assisted reproductive technologies outcome.

OBJECTIVE: To assess the association between recent cigarette smoking (CS) in female and male partners and assisted reproduction technology (ART) outcomes. DESIGN: Cohort prospective study. SETTING: University ART program in Chile. PATIENT(S): One hundred sixty-six couples seeking pregnancy through ART. INTERVENTION(S): Follicular fluid (FF) and serum cotinine concentrations were measured in female partners. Self-reported CS data were collected through personal interviews. MAIN OUTCOME MEASURE(S): The association between female recent smoking, assessed by FF and serum cotinine concentrations, and ART outcomes, such as number of ova retrieved and implantation rates, and the association between self-reported male recent smoking and live birth rates. RESULT(S): A significant age-adjusted association between increased FF cotinine level and decreased number of ova retrieved was found. The male partner's smoking habit significantly decreased the live birth rate from 21.1% to 7.8%. Serum cotinine concentrations paralleled those of FF. CONCLUSION(S): The hypothesis of a detrimental effect of recent female smoking over implantation rates is rejected. However, recent male smoking is associated with significantly decreased live birth rates even after adjusting for confounders. Female recent smoking was significantly associated with decreased number of retrieved ova.

Selection and identification of bacterial strains with methyl-tert-butyl ether, ethyl-tert-butyl ether, and tert-amyl methyl ether degrading capacities.

Nine bacterial strains isolated from two hydrocarbon-contaminated soils were selected because of their capacity for growth in culture media amended with 200 mg/L of one of the following gasoline oxygenates: Methyl-tert-butyl ether (MTBE), ethyl-tert-butyl ether (ETBE), and tert-amyl methyl ether (TAME). These strains were identified by amplification of their 16S rRNA gene, using fDl and rD1 primers, and were tested for their capacity to grow and biotransform these oxygenates in both mineral and cometabolic media. The isolates were classified as Bacillus simplex, Bacillus drentensis, Arthrobacter sp., Acinetobacter calcoaceticus, Acinetobacter sp., Gordonia amicalis (two strains), Nocardioides sp., and Rhodococcus ruber. Arthrobacter sp. (strain MG) and A. calcoaceticus (strain M10) consumed 100 (cometabolic medium) and 82 mg/L (mineral medium) of oxygenate TAME in 21 d, respectively, under aerobic conditions. Rhodococcus ruber (strain E10) was observed to use MTBE and ETBE as the sole carbon and energy source, whereas G. amicalis (strain T3) used TAME as the sole carbon and energy source for growth. All the bacterial strains transformed oxygenates better in the presence of an alternative carbon source (ethanol) with the exception of A. calcoaceticus (strain M10). The capacity of the selected strains to remove MTBE, ETBE, and TAME looks promising for application in bioremediation technologies.

Prevention of trocar site hernias: description of the safe port plug technique and preliminary results.

The aim of this study was to describe a new technique to prevent the development of incisional hernias in trocar sites. Between March and June 2006, a pilot study was conducted to determine the feasibility of the safe port plug technique using the Bioabsorbable Hernia Plug to prevent incisional hernia in trocar sites. The device was implanted in the umbilical trocar site (10-11 mm) of 17 patients undergoing laparoscopic surgery during the study period. The mean follow-up of patients was 14.6 months. Implantation of the Bioabsorbable Hernia Plug device by the safe port plug technique was possible in all cases. No patient presented complications in the follow-up. Our preliminary experience suggests that this technique is simple and feasible, and we hypothesized that this technique could be superior to conventional fascial closure: a hypothesis that must be proven in a randomized prospective trial that is currently in progress.

Gene expression in diplosporous and sexual Eragrostis curvula genotypes with differing ploidy levels.

The molecular nature of gene expression during the initiation and progress of diplosporous apomixis is still unknown. Moreover, the basis of the close correlation between diplospory and polyploidy is not clarified yet. A comparative expression analysis was performed based on expressed sequence tags (ESTs) sequencing and differential display in an Eragrostis curvula diplosporous tetraploid genotype (T, 4x apo), a sexual diploid derivative obtained from tissue culture (D, 2x sex) and an artificial sexual tetraploid obtained from the diploid seeds after colchicine treatment (C, 4x sex). From a total of 8,884 unigenes sequenced from inflorescence-derived libraries, 112 (1.26%) showed significant differential expression in individuals with different ploidy level and/or variable reproductive mode. Independent comparisons between plants with different reproductive mode (same ploidy) or different ploidy level (same reproductive mode) allowed the identification of genes modulated in response to diplosporous development or polyploidization, respectively. Surprisingly, a group of genes (Group 3) were differentially expressed or silenced only in the 4x sex plant, presenting similar levels of expression in the 4x apo and the 2x sex genotypes. A group of randomly selected differential genes was validated by QR-PCR. Differential display analysis showed that in general the 4x apo and 4x sex expression profiles were more related and different from the 2x sex one, but confirmed the existence of Group 3-type genes, in both inflorescences and leaves. The possible biological significance for the occurrence of this particular group of genes is discussed. In silico mapping onto the rice genome was used to identify candidates mapping to the region syntenic to the diplospory locus.

Division of the small bowel with the LigaSure Atlas device during the right laparoscopic colectomy.

BACKGROUND: Bowel division and anastomosis were facilitated greatly with the advent of stapling techniques. Since then, there have not been any new evolving technologies to facilitate these maneuvers. For this reason, we recently applied the LigaSure Atlas device (Valleylab, Boulder, CO) to the division of the small bowel during this procedure to obtain a reliable and low-cost division of the ileum. METHODS: After vessel ligation and ileocolic mobilization, the terminal ileum is sealed and divided with the LigaSure Atlas. This device is applied sequentially along the small bowel twice to ensure an adequate seal before the cut. A terminolateral ileotransversostomy is performed extracorporeally with a mechanical circular stapling device, installing the anvil of the circular stapler into the ileal stump. RESULTS: We have not encountered any problems with this technique since its introduction in our institution, and no leakage or bursting of the ileal stumps. CONCLUSIONS: This technique enables an easy, reliable, and inexpensive technical option to optimize the right laparoscopic colectomy procedure.

Bacillus farraginis sp. nov., Bacillus fortis sp. nov. and Bacillus fordii sp. nov., isolated at dairy farms.

Forty-eight bacterial strains were isolated at dairy farms from raw milk, the milking apparatus, green fodder or feed concentrate after a heat treatment of 30 min at 100 degrees C. In this way, spore-forming bacteria with a very high intrinsic heat resistance were selected for. The aerobic spore-forming isolates were subjected to a polyphasic taxonomical study, including repetitive element sequence-based PCR typing, whole-cell protein profiling, 16S rDNA sequence analysis, DNA-DNA hybridizations, DNA base composition, fatty acid analysis, and morphological and biochemical characteristics. A comparison of the REP- and (GTG)5-PCR and whole-cell protein SDS-PAGE profiles resulted in three clusters of similar strains. Analysis of the 16S rDNA sequences and DNA-DNA relatedness data showed that these clusters represented three novel species. The highest 16S rDNA similarity to a recognized species found for the three groups was around 94% with Bacillus lentus and Bacillus sporothermodurans. Further phenotypic characterization supported the proposal of three novel species in the genus Bacillus, Bacillus farraginis, Bacillus fortis and Bacillus fordii. The respective type strains are R-6540T (=LMG 22081T=DSM 16013T), R-6514T (=LMG 22079T=DSM 16012T) and R-7190T (=LMG 22080T=DSM 16014T); their G+C DNA base contents are 43.7, 44.3 and 41.9 mol%, respectively. Although in variable amounts, a predominance of the branched fatty acids iso-C(15 : 0) and anteiso-C(15 : 0) was observed in all three novel species.

Polyphasic characterization of Bacillus coagulans strains, illustrating heterogeneity within this species, and emended description of the species.

Because of its food spoiling capacity on the one hand and its significant role in the production of industrially valuable products on the other, Bacillus coagulans is of economic concern. Several studies have revealed a great deal of diversity within the species and this has led to a number of taxonomic adjustments. The present study aims to clarify the diversity within Bacillus coagulans sensu stricto and determine the taxonomic status of the species. Therefore, a polyphasic study was performed on a set of B. coagulans strains from diverse habitats. Techniques as ARDRA, SDS-PAGE of whole cell proteins, FAME analysis, routine phenotypic tests and rep-PCR illustrate considerable intra-species heterogeneity, while 16S rDNA sequence comparison and DNA-DNA relatedness support the accommodation of these strains in one species. Although most techniques demonstrate appreciable heterogeneity among the Bacillus coagulans strains, the intraspecies groupings are not consistent throughout all the methods applied and are not supported by any economic, historic or practical traits. Therefore, a division in subspecies seems inappropriate. In attempt to achieve a better species delineation, an emended description of Bacillus coagulans is included.

Bacillus novalis sp. nov., Bacillus vireti sp. nov., Bacillus soli sp. nov., Bacillus bataviensis sp. nov. and Bacillus drentensis sp. nov., from the Drentse A grasslands.

A group of 42 isolates were isolated from the soil of several disused hay fields, in the Drentse A agricultural research area (The Netherlands), that were taken out of production at different times. The group represents hitherto-uncultured Bacillus lineages that have previously been found, by a non-cultural method, to be predominant in soil. The strains were subjected to a polyphasic taxonomic study, including (GTG)5-PCR, 16S rDNA sequence analysis, DNA-DNA hybridizations, DNA base-ratio determination, fatty acid analysis and morphological and biochemical characterization. By comparing the groupings obtained by (GTG)5-PCR and 16S rDNA sequence analysis, six clusters of similar strains could be recognized. A DNA-DNA relatedness study showed that these clusters represented five novel genospecies. Further analysis supported the proposal of five novel species in the genus Bacillus, namely Bacillus novalis sp. nov. (type strain IDA3307T=R-15439T=LMG 21837T=DSM 15603T), Bacillus vireti sp. nov. (type strain IDA3632T=R-15447T=LMG 21834T=DSM 15602T), Bacillus soli sp. nov. (type strain IDA0086T=R-16300T=LMG 21838T=DSM 15604T), Bacillus bataviensis sp. nov. (type strain IDA1115T=R-16315T=LMG 21833T=DSM 15601T) and Bacillus drentensis sp. nov. (type strain IDA1967T=R-16337T=LMG 21831T=DSM 15600T).