Effect of an intramammary lipopolysaccharide challenge on the hindgut microbial composition and fermentation of dairy cattle experiencing intermittent subacute ruminal acidosis.

Feeding grain-rich diets often results in subacute ruminal acidosis (SARA), a condition associated with ruminal dysbiosis and systemic inflammation. Yet, the effect of SARA on hindgut microbiota, and whether this condition is aggravated by exogenous immune stimuli, is less understood. Therefore, the aims of this study were to determine the effects of an intermittent high-grain SARA model on the hindgut microbial community, and to evaluate whether the effects of SARA on the fecal microbiome and fermentation were further affected by an intramammary lipopolysaccharide (LPS) challenge. A total of 18 early-lactating Simmental cows were divided into 3 groups (n = 6); 2 were fed a SARA-inducing feeding regimen (60% concentrate), 1 was fed a control (CON) diet (40% concentrate). On d 30, 1 SARA group (SARA-LPS) and the CON group (CON-LPS) were intramammarily challenged with a single dose of 50 µg of LPS from Escherichia coli O26:B6, whereas the remaining 6 SARA cows (SARA-PLA) received a placebo. Using a longitudinal randomized controlled design, with grouping according to parity and days in milk), statistical analysis was performed with baseline measurements used as a covariate in a mixed model procedure. The SARA-inducing feeding challenge resulted in decreased fecal pH and increased butyrate as a proportion of total short-chain fatty acids in the feces. On d 30, SARA-challenged cows had decreased fecal diversity as shown by the Shannon and Chao1 indices and a decrease in the relative abundance of Euryarchaeota and cellulolytic genera, and numerical increases in the relative abundance of several Firmicutes associated with starch and secondary fermentation. The LPS challenge did not affect the fecal pH and short-chain fatty acids, but increased the Chao1 richness index in an interaction with the SARA challenge, and affected the relative abundance of Verrucomicrobia (1.13%), Actinobacteria (0.19%), and Spirochaetes (0.002%), suggesting an effect on the microbial ecology of the hindgut during SARA conditions. In conclusion, the SARA-inducing feeding regimen promoted important microbial changes at d 30, including reduced diversity and evenness compared with CON, whereas the external LPS challenge led to changes in the microbial community without affecting fecal fermentation properties.

Adaptive responses in short-chain fatty acid absorption, gene expression, and bacterial community of the bovine rumen epithelium recovered from a continuous or transient high-grain feeding.

The feeding of high-grain diets to dairy cows commonly results in lowered pH and ruminal dysbiosis, characterized by changes in absorption dynamics of short-chain fatty acids (SCFA) across the reticuloruminal wall, epithelial function, and the epithelial bacteria community structure. Therefore, the present study evaluated the effect of high-grain feeding persistence on the absorption kinetics of reticuloruminal SCFA, gene expression in the rumen epithelium, and the associated shifts in the epithelial bacteria in cows recovering from either a long-term continuous high-grain feeding model or a long-term transient high-grain feeding model. In a crossover study design, 8 nonlactating Holstein cows were fed 60% concentrate either continuously for 4 wk (continuous) or with a 1-wk break in the second week of the high-grain feeding (transient). After the high-grain feeding, all animals were fed a diet of 100% forage (recovery) for an additional 8 wk. Rumen papilla biopsies and SCFA absorption measurements were taken at the start of the trial (baseline), after the 4-wk high-grain feeding (49 d), after 2-wk recovery forage feeding (63 d), and after 8-wk recovery forage (105 d). Absorption of SCFA was determined in vivo using the washed and isolated reticulorumen technique. Rumen papillae biopsies were used for adherent bacterial DNA and host RNA extraction. The epithelial bacteria were determined using Illumina MiSeq (Microsynth AG, Balgach, Switzerland) sequencing of the 16S rRNA gene. No significant effects of the high-grain feeding model were seen for bacterial diversity. However, bacterial diversity increased with time spent in the recovery forage feeding period regardless of feeding model. The relative abundance of Acidobacteria phyla and Acetivibrio spp. increased when animals were fed a transient high-grain feeding model. A trend toward increased CLDN4 expression was observed in the continuous model. Furthermore, there were interactions between feeding model and sampling day for gene targets CD14, DRA, NHE2, NHE3, and MCT2. When comparing length of recovery, in the continuous model increased relative absorption of SCFA was sustained at 63 d but dropped to baseline measurements at 105 d. A similar pattern was found with the transient model but it did not reach significance. The only gene target that was found to significantly correlate to relative absorption of SCFA was DRA (correlation coefficient ≤ -0.41). Whereas, genera Alkalibaculum, Anaerorhabdus, Coprococcus, and Dethiobacter all showed positive correlations to gene targets for pH regulation (NHE2 and NHE3) and SCFA uptake (MCT1) but negative correlations to SCFA absorption. We conclude that while the rumen absorption and epithelial bacteria were able to recover to baseline levels after 8 wk of forage feeding, the time needed for re-establishment of homeostasis in host gene expression is longer, especially when high-grain feeding is interrupted.

Changes in the Rumen Epithelial Microbiota of Cattle and Host Gene Expression in Response to Alterations in Dietary Carbohydrate Composition.

The inclusion of high-quality hay (HQH), in place of concentrates, shifts dietary carbohydrate intake, and the extent to which these shifts effect epimural microbiota and epithelial gene expression of the rumen has not yet been evaluated. Eight ruminally cannulated nonlactating Holstein cows were used in a replicated 4 by 4 Latin square design with four dietary treatments containing HQH, with either 0% concentrate/100% HQH (100HQH), 25% concentrate/75% HQH (75HQH), or 40% concentrate/60% HQH (60HQH). The fourth group (control [CON]) was fed 60% normal fiber-rich hay and 40% concentrate. The data showed that measures of diversity for the rumen epimural population, specifically the Shannon (P = 0.004) and Simpson (P = 0.003) indices, decreased with increasing levels of HQH in the diet. The feeding of HQH shifted the epimural population from predominantly Firmicutes to Proteobacteria Phylogenetic analysis revealed that HQH feeding markedly shifted the abundance of Campylobacter spp. from 7.8 up to 33.5% (P < 0.001), with greater ingestion of protein (r = 0.63) and sugars (r = 0.65) in HQH diet being responsible for this shift. The expression of genes targeting intracellular pH regulation, barrier function, and nutrient uptake of rumen epithelium remained stable regardless of the carbohydrate source. In conclusion, the data suggest strong alterations of the ruminal epimural microbiota in response to changes in the nutritive patterns of the diet. Further research is warranted to evaluate the long-term effects of these significant microbial changes on rumen health and food safety aspects in cattle at a transcriptional level.IMPORTANCE Feeding of forages versus starchy concentrates is a highly debated topic. Hay is believed to be healthier and more ecological sustainable for cattle than are concentrates, although the effects of feeding hay with enhanced sugar and protein content on epimural microbiota and host gene expression have not yet been evaluated. This research provides a report of the role of feeding hay with increased sugar and protein content in place of starchy concentrates in altering epimural microbiota and in generating a host response. Our research shows that the addition of high-quality hay to dairy rations shifted nutrient intake, resulting in strong alterations in the epimural microbiota in cattle. This work provides a background for further long-term research regarding the effects of feeding practices on the host-microbiome interaction and its role in rumen health and food safety in cattle.

Graded substitution of grains with bakery by-products modulates ruminal fermentation, nutrient degradation, and microbial community composition in vitro.

A new segment of feed industry based on bakery by-products (BBP) has emerged. Yet, information is lacking regarding the effects of inclusion of BBP in ruminant diets on ruminal fermentation and microbiota. Therefore, the aim of this study was to evaluate the effect of the gradual replacement of grains by BBP on ruminal fermentation, nutrient degradation, and microbial community composition using the rumen-simulation technique. All diets consisted of hay and concentrate mixture with a ratio of 42:58 (dry matter basis), but differed in the concentrate composition with either 45% cereal grains or BBP, whereby 15, 30, or 45% of BBP were used in place of cereal grains. The inclusion of increasing levels of BBP in the diet linearly enhanced ruminal degradation of starch from 84% (control) to 96% (45% BBP), while decreasing degradation of crude protein and fiber. The formation of methane was lowered in the 45% BBP diet compared with all other diets. Whereas the ammonia concentration was similar in the control and 15% BBP, a significant decrease was found in 30% BBP (-23%) and 45% BBP (-33%). Also, BBP feeding shifted fermentation profile toward propionate at the expense of acetate. Moreover, isobutyrate linearly decreased with increasing BBP inclusion. Bacterial 16S rRNA Illumina MiSeq (Microsynth AG, Balach, Switzerland) sequencing revealed a decreased microbial diversity for the 45% BBP diet. Furthermore, the replacement of cereal grains with BBP went along with an increased abundance of the genera Prevotella, Roseburia, and Megasphaera, while decreasing Butyrivibrio and several OTU belonging to Ruminococcaceae. In conclusion, the inclusion of BBP at up to 30% of the dry matter had no detrimental effects on pH, fiber degradability, and microbial diversity, and enhanced propionate production. However, a higher replacement level (45%) impaired ruminal fermentation traits and fiber degradation and is not recommended.

High-grain diets supplemented with phytogenic compounds or autolyzed yeast modulate ruminal bacterial community and fermentation in dry cows.

The feeding of concentrate-rich diets may lead to microbial imbalances and dysfermentation in the rumen. The main objective of this study was to determine the effects of supplementing phytogenic compounds (PHY) or autolyzed yeast (AY) on rumen fermentation and microbial abundance in cows intermittently fed concentrate-rich diets. The experiment was carried out as an incomplete 3 × 4 Latin square design, with 8 nonlactating rumen-fistulated Holstein-Friesian cows. The cows were randomly assigned to a concentrate diet that was either not supplemented (CON), or supplemented with PHY or AY. Each of the 4 consecutive experimental periods was composed of a 1-wk roughage-only diet (RD), 6-d gradual concentrate increase, followed by 1 wk of 65% concentrate (dry matter basis; Conc I), and 1 wk of RD and a final 2-wk 65% concentrate (dry matter basis; Conc II) phase. Digesta samples were collected from the rumen mat for bacterial 16S rRNA gene Illumina MiSeq (Illumina, Balgach, Switzerland) sequencing, and samples of particle-associated rumen liquid were obtained for measuring short-chain fatty acids, lactate, ammonia, and pH during RD (d 6), Conc I (d 19), and Conc II (d 39). The concentrate feeding caused a decrease of overall bacterial diversity indices, especially during Conc I. The genera Ruminococcus, Butyrivibrio, and Coprococcus were decreased, whereas Prevotella, Megasphaera, Lachnospira, and Bacteroides were increased in abundance. Supplementation of both feed additives increased the abundance of gram-positive and decreased that of gram-negative bacteria. Supplementation of AY enhanced cellulolytic bacteria such as Ruminococcus spp., whereas PHY decreased starch and sugar fermenters including Bacteroides spp., Shuttleworthia spp., and Syntrophococcus spp. Moreover, PHY supplementation increased butyrate percentage in the rumen in both concentrate phases. In conclusion, intermittent high-concentrate feeding altered the digesta-associated rumen bacterial community and rumen fermentation with more significant alterations found in Conc I than in Conc II. The data also showed that both feed additives had the most significant modulatory effects on the bacterial community, and their subsequent fermentation, during periods of low pH.

Invited review: Practical feeding management recommendations to mitigate the risk of subacute ruminal acidosis in dairy cattle.

Rumen health is of vital importance in ensuring healthy and efficient dairy cattle production. Current feeding programs for cattle recommend concentrate-rich diets to meet the high nutritional needs of cows during lactation and enhance cost-efficiency. These diets, however, can impair rumen health. The term "subacute ruminal acidosis" (SARA) is often used as a synonym for poor rumen health. In this review, we first describe the physiological demands of cattle for dietary physically effective fiber. We also provide background information on the importance of enhancing salivary secretions and short-chain fatty acid absorption across the stratified squamous epithelium of the rumen; thus, preventing the disruption of the ruminal acid-base balance, a process that paves the way for acidification of the rumen. On-farm evaluation of dietary fiber adequacy is challenging for both nutritionists and veterinarians; therefore, this review provides practical recommendations on how to evaluate the physical effectiveness of the diet based on differences in particle size distribution, fiber content, and the type of concentrate fed, both when the latter is part of total mixed ration and when it is supplemented in partial mixed rations. Besides considering the absolute amount of physically effective fiber and starch types in the diet, we highlight the role of several feeding management factors that affect rumen health and should be considered to control and mitigate SARA. Most importantly, transitional feeding to ensure gradual adaptation of the ruminal epithelium and microbiota; monitoring and careful management of particle size distribution; controlling feed sorting, meal size, and meal frequency; and paying special attention to primiparous cows are some of the feeding management tools that can help in sustaining rumen health in high-producing dairy herds. Supplementation of feed additives including yeast products, phytogenic compounds, and buffers may help attenuate SARA, especially during stress periods when the risk of a deficiency of physically effective fiber in the diet is high, such as during early lactation. However, the usage of feed additives cannot fully compensate for suboptimal feeding management.

Isolated axillary nodal swelling and cancer of unknown primary.

INTRODUCTION: The literature reports rare cases of isolated axillary lymph node metastasis from cancer of unknown primary (CUP). The authors reviewed the prevalence and outcome of patients with isolated axillary nodal swelling suspicious for malignancy affected or not by isolated axillary node metastasis from CUP. MATERIALS AND METHODS: The authors collected data about 65 patients presented with isolated axillary lymph node swelling who underwent axillary lymph node excisional biopsy for malignancy suspicion, between January 2005 and December 2011, in the absence of any specific diagnosis. RESULTS: Histological examination revealed a metastatic infiltration by an occult solid cancer in 16 cases (24%), ten of which were occult breast cancers. Histological patterns and molecular markers allowed in all cases of occult cancer a probable identification of the primary tumor site, while a definitive diagnosis was possible only in the 56.25% of cases (9/16). The prognosis of these patients was very poor with a five-year overall survival of 28%, and thus very similar to patients affected by Stage IV overt breast cancer. CONCLUSIONS: Among occult malignancies presenting with sole axillary lymph node metastasis, breast cancer remains the more probable primary cancer, but many other sites should be taken into consideration by negative breast imaging. Positron-emission tomography computed tomography (PET-CT) resulted helpful in the primary site detection, but has nonetheless a margin of failure. Occult breast cancers behave very similar to Stage IV overt breast cancers, and should be treated accordingly.

Laparoscopic peritoneal lavage for perforated colonic diverticulitis: a definitive treatment? Retrospective analysis of 63 cases.

BACKGROUND: The ideal treatment of perforated diverticulitis and the indications for elective colon resection remain controversial. Considering the significant morbidity and mortality rates related to traditional resection, efforts have been made to reduce the invasiveness of surgery in recent decades. Laparoscopic peritoneal lavage has emerged as an effective alternative option. We retrospectively investigated the effectiveness of laparoscopic peritoneal lavage for perforated diverticulitis and the possibility that it could be a definitive treatment. METHODS: We included patients treated with laparoscopic peritoneal lavage for perforated diverticulitis. The inclusion criteria were all emergency patients with generalized peritonitis due to Hinchey III perforated diverticulitis and some cases of Hinchey II and IV. RESULTS: Sixty-three patients were treated with laparoscopic peritoneal lavage. Six patients (9.5 %) had Hinchey II diverticulitis; 54 patients (85.7 %) had Hinchey III; and three patients (4.8 %) had Hinchey IV. The mean operative time was 87.3 min (±25.4 min), and the overall morbidity rate was 14.3 %. One patient died because of pulmonary embolism, and there were six early reinterventions because of treatment failure. Delayed colon resection was performed in four of the remaining 57 patients (7 %) because of recurrent diverticulitis. In the other 53 patients (93 %), we saw no recurrence of diverticulitis and no intervention was performed after a median follow-up period of 54 months (interquartile range 27-98 months). CONCLUSIONS: Laparoscopic peritoneal lavage for perforated diverticulitis can be considered a safe and effective alternative to traditional surgical resection, and using this approach, most elective colon resection might be avoided.

Changes in the rumen epimural bacterial diversity of beef cattle as affected by diet and induced ruminal acidosis.

Little is known about the nature of the rumen epithelial adherent (epimural) microbiome in cattle fed different diets. Using denaturing gradient gel electrophoresis (DGGE), quantitative real-time PCR (qPCR), and pyrosequencing of the V3 hypervariable coding region of 16S rRNA, epimural bacterial communities of 8 cattle were profiled during the transition from a forage to a high-concentrate diet, during acidosis, and after recovery. A total of 153,621 high-quality gene sequences were obtained, with populations exhibiting less taxonomic variability among individuals than across diets. The bacterial community composition exhibited clustering (P < 0.03) by diet, with only 14 genera, representing >1% of the rumen epimural population, differing (P ≤ 0.05) among diets. During acidosis, levels of Atopobium, Desulfocurvus, Fervidicola, Lactobacillus, and Olsenella increased, while during the recovery, Desulfocurvus, Lactobacillus, and Olsenella reverted to levels similar to those with the high-grain diet and Sharpea and Succinivibrio reverted to levels similar to those with the forage diet. The relative abundances of bacterial populations changed during diet transition for all qPCR targets except Streptococcus spp. Less than 5% of total operational taxonomic units (OTUs) identified exhibited significant variability across diets. Based on DGGE, the community structures of epithelial populations differed (P ≤ 0.10); segregation was most prominent for the mixed forage diet versus the grain, acidotic challenge, and recovery diets. Atopobium, cc142, Lactobacillus, Olsenella, RC39, Sharpea, Solobacterium, Succiniclasticum, and Syntrophococcus were particularly prevalent during acidosis. Determining the metabolic roles of these key genera in the rumens of cattle fed high-grain diets could define a clinical microbial profile associated with ruminal acidosis.

New surgical trends in breast cancer treatment: conservative interventions and oncoplastic breast surgery.

AIM: From radical mastectomy by Halstedt to quadrantectomy by Veronesi, surgical treatment of breast cancer has become more and more conservative in order to preserve women quality of life. We analyzed the evolution of breast interventions through a nine-year period. METHODS: We collected data about all breast surgery interventions performed between 2002 and 2010 in our Department Of Surgery, focusing on patients' age, tumor histology, TNM classification, hormonal status, intervention radicality on the breast and axilla, and eventual plastic surgery. Data were analyzed by R (version 2.14.2), considering significant P<0.05. RESULTS: A total of 3320 breast surgery interventions were performed on 2300 patients. Absolute yearly number of interventions has strongly increased with the introduction of the mammography screening (291 to 430). Conservative breast surgery (55% to 62%) and skin-sparing mastectomy (1% to 8%) have been performed more frequently in premenopausal patients, and significantly increased in time, with a consequent decrease of classical radical mastectomy (38 to 15%) but an increase of margins widening after primary surgery (2% to 6%). Sentinel lymph node biopsy is mostly replacing complete axillary lymph node dissection (93% to 31%). Skin-glandular reshaping progressively increased (up to 20%) as also breast reconstruction (23% to 40%), which since 2008 has been performed also in women older than 75 years, while flaps have been used in strictly selected patients. CONCLUSION: Conservative surgery represents the target of current breast cancer treatment where possible, and skin-sparing mastectomy an interesting alternative to classical one when radicality is required. Sentinel lymph node biopsy has replaced complete axillary lymph node dissection by clinically negative nodal status. Breast reshaping and reconstruction are increasing in every age group, including women older than 75 years.

Characterization of rumen bacterial diversity and fermentation parameters in concentrate fed cattle with and without forage.

AIMS: To determine the effects of the removal of forage in high-concentrate diets on rumen fermentation conditions and rumen bacterial populations using culture-independent methods. METHODS AND RESULTS: Detectable bacteria and fermentation parameters were measured in the solid and liquid fractions of digesta from cattle fed two dietary treatments, high concentrate (HC) and high concentrate without forage (HCNF). Comparison of rumen fermentation conditions showed that duration of time spent below pH 5·2 and rumen osmolality were higher in the HCNF treatment. Simpson's index of 16S PCR-DGGE images showed a greater diversity of dominant species in the HCNF treatment. Real-time qPCR showed populations of Fibrobacter succinogenes (P = 0·01) were lower in HCNF than HC diets. Ruminococcus spp., F. succinogenes and Selenomonas ruminantium were at higher (P ≤ 0·05) concentrations in the solid vs the liquid fraction of digesta regardless of diet. CONCLUSIONS: The detectable bacterial community structure in the rumen is highly diverse. Reducing diet complexity by removing forage increased bacterial diversity despite the associated reduction in ruminal pH being less conducive for fibrolytic bacterial populations. Quantitative PCR showed that removal of forage from the diet resulted in a decline in the density of some, but not all fibrolytic bacterial species examined. SIGNIFICANCE AND IMPACT OF THE STUDY: Molecular techniques such as DGGE and qPCR provide an increased understanding of the impacts of dietary changes on the nature of rumen bacterial populations, and conclusions derived using these techniques may not match those previously derived using traditional laboratory culturing techniques.

Breast-feeding and benign breast disease.

Benign breast disease (BBD) is very common among women in their fertile age, but its correlation with breast reproductive function remains unclear. Our study aimed to investigate the relation between BBD and breast-feeding. We collected data on 105 women with BBD and 98 controls, focusing on their reproductive history and breast-feeding. We analysed data by R (version 2.12.1) considering p < 0.05 as significant. The results showed that fibroadenoma represented the most frequent BBD (55%), followed by fibrocystic changes (19%), intraductal papilloma (6%) and inflammatory breast disorders (5%). The mean age was 31.5 years (± 6.1), BMI 21.2 kg/m² (± 3.4) and age at menarche 13.0 years (± 1.5). Duration of breast-feeding was not significantly different between controls and BBD types (p = NS). Selecting women with fibroadenoma breast-feeding duration directly correlated with the number of benign lesions (p < 0.05), which remains significant also by multivariate analysis. It was concluded that there seemed to be no difference in breast-feeding among BBDs types, but lactation may influence the number of fibroadenomas. Moreover, prospective studies would better define the correlation between lactation and BBDs.

Corrigendum to 'A dataset composed of multiangular spectral libraries and auxiliary data at tree, leaf, needle, and bark level for three common European tree species' [Data in Brief 35 (2021) 106820].

[This corrects the article DOI: 10.1016/j.dib.2021.106820.].

A dataset composed of multiangular spectral libraries and auxiliary data at tree, leaf, needle, and bark level for three common European tree species.

This article describes a dataset of multiangular scattering properties of small trees (height = 0.38-0.7 m) at visible, near-infrared, and shortwave-infrared wavelengths (350-2500 nm), and provides supporting auxiliary data that comprise leaf, needle, and bark spectra, and structural characteristics of the trees. Multiangular spectra were measured for 18 trees belonging to three common European tree species: Scots pine (Pinus sylvestris L.), Norway spruce (Picea abies (L.) H. Karst), and sessile oak (Quercus petraea (Matt.) Liebl.). The measurements were performed in 47 different view angles across a hemisphere, using a laboratory goniometer and a non-imaging spectrometer. Leaf and needle spectra were measured for each tree, using a non-imaging spectrometer coupled to an integrating sphere. Bark spectra were measured for one sample tree per species. In addition, leaf and needle fresh mass, surface area of leaves, needles, and woody parts, silhouette area, and spherically averaged silhouette to total area ratio (STAR) for each tree were measured or derived from the measurements. The data are useful for modeling the shortwave reflectance characteristics of small trees and potentially forests, and thus benefit climate modeling or interpretation of remote sensing data.

Contribution of indirect computed tomography venography to computed tomography angiography of the chest for the diagnosis of thromboembolic disease in two United States emergency departments.

Recent reports suggest that physicians in non-ambulatory settings can use indirect CT venography (CTV) of the lower extremities immediately following spiral CT angiography (CTA) of the chest to identify patients with a negative CTA who have thromboembolic disease identified on CTV. We sought to determine the frequency of isolated deep venous thrombosis (DVT) discovered on CTV in emergency department (ED) patients with complaints suggestive of pulmonary embolism (PE) yet having a negative CTA. This study was conducted in a suburban and urban ED where patients with symptoms suspicious for PE were primarily evaluated with CTA and CTV. A total of 800 patients were studied, including 360 from the suburban ED and 440 from the urban ED. 88 (11%) patients were diagnosed with thromboembolic disease by CTA, or CTV, or both. Seventy-three patients had a CTA of the chest that was positive for PE, 42 (5.2%) of whom had evidence of both PE on CTA and DVT on CTV. Fifteen patients (2%, 95% CI = 1-3%) had a negative CTA and were subsequently found to have isolated DVT on CTV, all of whom received anticoagulation therapy. These data suggest that indirect CT venography of immediately following CT angiography of the chest significantly increased the frequency of diagnosed thromboembolic disease requiring anticoagulation in ED patients with suspected PE.

Phylogeny and distribution of the soxB gene among thiosulfate-oxidizing bacteria.

A PCR protocol for the detection of sulfur-oxidizing bacteria based on soxB genes that are essential for thiosulfate oxidation by sulfur-oxidizing bacteria of various phylogenetic groups which use the 'Paracoccus sulfur oxidation' pathway was developed. Five degenerate primers were used to specifically amplify fragments of soxB genes from different sulfur-oxidizing bacteria previously shown to oxidize thiosulfate. The PCR yielded a soxB fragment of approximately 1000 bp from most of the bacteria. Amino acid and nucleotide sequences of soxB from reference strains as well as from new isolates and environmental DNA from a hydrothermal vent habitat in the North Fiji Basin were compared and used to infer relationships of soxB between sulfur-oxidizing bacteria belonging to various 16S rDNA-based phylogenetic groups. Major phylogenetic lines derived from 16S rDNA were confirmed by soxB phylogeny. Thiosulfate-oxidizing green sulfur bacteria formed a coherent group by their soxB sequences. Likewise, clearly separated branches demonstrated the distant relationship of representatives of alpha-, beta-, and gamma-Proteobacteria including representative species of the former genus Thiobacillus (now Halothiobacillus - gamma-Proteobacteria, Thiobacillus - beta-Proteobacteria and Starkeya - alpha-Proteobacteria). This general picture emerged although apparent evidence for lateral transfer of the soxB gene is indicated and comparison of soxB phylogeny and 16S rDNA phylogeny points to the significance of this gene transfer in hydrothermal vent bacterial communities of the North Fiji Basin.

The relationship of nitrate reducing bacteria on the basis of narH gene sequences and comparison of narH and 16S rDNA based phylogeny.

On the basis of available nitrate reductase gene sequences primer pairs were designed to specifically amplify gene stretches of the beta-subunit of the membrane-bound nitrate reductase (narH). Additional sequences of this gene were amplified and sequenced from pure cultures of reference species and new isolates. The distribution and phylogeny of this gene in denitrifying and nitrate-reducing bacteria was analysed. Comparison of phylogenetic trees based on 16S rDNA sequences with those based on narH sequences revealed highly similar relationships of both genes from most of the bacteria analysed. Since highly conserved functional cysteine clusters within bacterial and archaeal narH sequences support a linear evolution from one common progenitor a long evolutionary history of the respiratory membrane-bound nitrate reductase can be inferred from our phylogenetic data.

Injury with spontaneous deployment of an automobile air bag.

We report a case of significant injury to an automobile driver due to spontaneous air bag deployment not associated with a collision or application of external force to the automobile.

Insertion of a straight peritoneal catheter in an arcuate subcutaneous tunnel by a tunneler: long-term experience.

OBJECTIVE: This study describes the results of the insertion of a straight Tenckhoff peritoneal catheter (PC) in an arcuate, caudally concave tunnel using a tunneler designed by the authors. It has a semicircular shape and a bending radius of 4.5 cm. SETTING: A hospital renal unit. PATIENTS: From June 1988 to February 1994, 112 straight Tenckhoff PCs, 62 with one deep cuff (single-cuff PC) and 50 with two cuffs (double-cuff PC), were inserted as first catheters in 112 patients (mean age 62 +/- 13 years), who underwent continuous ambulatory peritoneal dialysis (CAPD). The follow-up was 1099 months (mean 18 +/- 13 months) for single-cuff PCs and 1264 months (mean 25 +/- 15 months) for double-cuff PCs, respectively. INTERVENTIONS: After intraperitoneal placement of the PCs by median laparotomy, a 180 degrees arc bend tunnel, with both external and peritoneal exits directed downwards, was created by means of the tunneler. RESULTS: The rate of exit-site infection (ESI) was 0.27 episodes/year (epis/year). The probability of remaining ESI-free was 76%, 60%, and 55% at 1, 2, and 3 years. The rate of tunnel infection (TI) was 0.046 epis/year. The incidence of the double-cuff PC-related ESI and TI tended to be lower than the incidence observed with the single-cuff PC. Episodes of peritonitis were 60 (0.30 epis/year), where 6 were subsequent to ESI and/or TI. Two PCs were lost due to ESI, 3 due to TI, and 11 due to peritonitis. Drainage failure, due to displacement of the PC caused by straightening, involved 3 PCs; 2 were lost. PC survival was 92%, 82%, and 74% at 1, 2 and 3 years, respectively. CONCLUSIONS: By an easily used semicircular tunneler, the standard straight Tenckhoff PC can be stably positioned in an arcuate tunnel with both inner and outer exits directed downwards. This tunnel shape, as already suggested by some authors, appears to be an effective technical solution to reducing the PC-related complication rates.