Species diversity and substrate utilization patterns of thermophilic bacterial communities in hot aerobic poultry and cattle manure composts

This study investigated the species diversity and substrate utilization patterns of culturable thermophilic bacterial communities in hot aerobic poultry and cattle manure composts by coupling 16S rDNA analysis with Biolog data. Based on the phylogenetic relationships of 16S rDNA sequences, 34 thermophilic (grown at 60 degrees C bacteria isolated during aerobic composting of poultry manure and cattle manure were classified as Bacillus licheniformis, B. atrophaeus, Geobacillus stearothermophilus, G. thermodenitrificans, Brevibacillus thermoruber, Ureibacillus terrenus, U. thermosphaericus, and Paenibacillus cookii. In this study, B. atrophaeus, Br. thermoruber, and P. cookii were recorded for the first time in hot compost. Physiological profiles of these bacteria, obtained from the Biolog Gram-positive (GP) microplate system, were subjected to principal component analysis (PCA). All isolates were categorized into eight different PCA groups based on their substrate utilization patterns. The bacterial community from poultry manure compost comprised more divergent species (21 isolates, seven species) and utilized more diverse substrates (eight PCA groups) than that from cattle manure compost (13 isolates, five species, and four PCA groups). Many thermophilic bacteria isolated in this study could use a variety of carboxylic acids. Isolate B110 (from poultry manure compost), which is 97.6% similar to U. terrenus in its 16S rDNA sequence, possesses particularly high activity in utilizing a broad spectrum of substrates. This isolate may have potential applications in industry

Nicotine-activated descending facilitation on spinal NMDA-dependent reflex potentiation from pontine tegmentum in rats

This study was conducted to investigate the possible neurotransmitter that activates the descending pathways coming from the dorsolateral pontine tegmentum (DPT) to modulate spinal pelvic-urethra reflex potentiation. External urethra sphincter electromyogram (EUSE) activity in response to test stimulation (TS, 1/30 Hz) and repetitive stimulation (RS, 1 Hz) on the pelvic afferent nerve of 63 anesthetized rats were recorded with or without microinjection of nicotinic cholinergic receptor (nAChR) agonists, ACh and nicotine, to the DPT. TS evoked a baseline reflex activity with a single action potential (1.00 +/- 0.00 spikes/stimulation, n = 40), whereas RS produced a long-lasting reflex potentiation (16.14 +/- 0.96 spikes/stimulation, n = 40) that was abolished by D-2-amino-5-phosphonovaleric acid (1.60 +/- 0.89 spikes/stimulation, n = 40) and was attenuated by 2,3-dihydroxy-6-nitro-7sulfamoyl-benzo (F) quinoxaline (7.10 +/- 0.84 spikes/stimulation, n = 40). ACh and nicotine microinjections to DPT both produced facilitation on the RS-induced reflex potentiation (23.57 +/- 2.23 and 28.29 +/- 2.36 spikes/stimulation, P = 0.01, n = 10 and 20, respectively). Pretreatment of selective nicotinic receptor antagonist, chlorisondamine, reversed the facilitation on RS-induced reflex potentiation caused by nicotine (19.41 +/- 1.21 spikes/stimulation, P = 0.01, n = 10) Intrathecal WAY-100635 and spinal transection at the T(1) level both abolished the facilitation on reflex potentiation resulting from the DPT nicotine injection (12.86 +/- 3.13 and 15.57 +/- 1.72 spikes/stimulation, P < 0.01, n = 10 each). Our findings suggest that activation of nAChR at DPT may modulate N-methyl-D-aspartic acid-dependent reflex potentiation via descending serotonergic neurotransmission. This descending modulation may have physiological/pathological relevance in the neural controls of urethral closure

The ixodid ticks collected from dogs and other animals in Taiwan and Kinmen Island

A total of 594 ticks were collected from the environment, dogs, cattle, goats, small mammals, birds, a turtle, a snake, a bat and a human. Morphological identification of 509 ticks resulted in the listing of the following species: Rhipicephalus sanguineus, Rhipicephalus haemaphysaloides, Rhipicephalus microplus, Haemaphysalis warburtoni, Haemaphysalis phasiana, Haemaphysalis campanulata, Ixodes acutitarsus and Amblyomma geoemydae

Antibacterial activity of short hydrophobic and basic-rich peptides

Objective-To design short and potent analogs of bovine lactoferricin by use of the concepts of lipophilic bulk and cationic charge. Sample Population-5 synthetic peptides of bovine lactoferricin. Procedure-Antibacterial peptides were constructed by synthesizing several decapeptides rich in arginine and tryptophan. Basic residues of bovine lactoferricin (bLf 20-29; residues 20 to 29) were modified by substitution with arginine or lysine and nonbasic residues were modified by substitution with tryptophan, phenylalanine, or isoleucine. Synthetic peptides of bovine lactoferrin (LFB) were designated as LFB-RW (RRWWWRWRRW), LFB-KW (KKWWWKWKKW), LFB-RWa (RRWWRRWRRW), LFB-RF (RRFFFRFRRF), and LFB-RI (RRIIIRWRRI), where R, K, W, F, and I stand for arginine, lysine, tryptophan, phenylalanine, and isoleucine, respectively. Peptides were evaluated by determining their minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against Escherichia coli, Staphylococcus aureus, and Enterococcus faecalis. Results-LFB-RW, LFB-KW, and LFB-RWa possessed equivalent potency as bLf 20-29 against E coli. LFB-RW and LFB-RWa had a 2-fold increase in growth-inhibitory and bactericidal activity against S aureus, compared with bLf 20-29. LFB-RI had the lowest MIC value against E coli among the peptides but lost bactericidal activity. LFB-RW and LFB-KW had stronger bactericidal activities against S aureus or E faecalis, respectively, as well as E coli than the other synthetic peptides. LFB-RF also had antibacterial activity, but this was 2-fold less than that of LFB-RW, as determined by MIC and MBC values. Conclusions and Clinical Relevance-In construction of potent antibacterial peptides, inclusion of arginine, lysine, tryptophan, or isoleucine residues enhances effectiveness against certain bacteria, as measured by MIC or MBC values

Structural and biological characterization of mastoparans in the venom of Vespa species in Taiwan

Mastoparans, a family of small peptides, are isolated from the wasp venom. In this study, six mastoparans were identified in the venom of six Vespa species in Taiwan. The precursors of these mastoparans are composed of N-terminal signal sequence, prosequence, mature mastoparan, and appendix glycine at C-terminus. These mature mastoparans all have characteristic features of linear cationic peptides rich in hydrophobic and basic amino acids without disulfide bond. Therefore, these peptides could be predicted to adopt an amphipathic a-helical secondary structure. In fact, the CD (circular dichroism) spectra of these peptides show a high content a-helical conformation in the presence of 8 mM SDS or 40% 2,2,2-trifluoroethanol (TFE). All mastoparans exhibit mast cell degranulation activity, antimicrobial activity against both Gram-positive and -negative bacteria tested, various degree of hemolytic activity on chicken, human, and sheep erythrocytes as well as membrane permeabilization on Escherichia coil BL21. Our results also show that the hemolytic activity of mastoparans is correlated to mean hydrophobicity and mean hydrophobic moment. (C) 2011 Elsevier Inc. All rights reserved

Molecular cloning, expression, and functional characterization of a cystatin from pineapple stem

A cDNA fragment encoding the cysteine protease inhibitor, cystatin, was cloned from pineapple (Ananas comosus) stem. This clone was constructed in a fusion vector and was easily over-expressed in Escherichia coli; satisfactory over-expression of non-fusion cystatin was achieved after an additional start codon was inserted prior to its coding sequence. Both recombinant cystatins were predominately found in the soluble fraction of the cell extract, and were demonstrated to be functionally active in a reverse zymographic assay. The fusion and non-fusion cystatins were separately purified to homogeneity via a His-tag or papain-coupling affinity column. Effective inhibitory activity against papain was detected with both the fusion and non-fusion cystatins with comparable K-i values of 1.18 x 10(-10) m and 9.53 x 10(-11) (M), respectively. The recombinant cystatins were found to be thermally stable up to 60 degreesC. Inhibition of the endogenous protease activity in minced fish muscle revealed that the recombinant pineapple cystatins might be an adequate stabilizer to prevent protein degradation during industrial food processing

The Use of Commercial Soil Nucleic Acid Extraction Kit and Nested PCR for Detection of Leptospira in Farm Environment after Flooding in Taiwan

Leptospirosis is a rapidly transmitted and flood-related disease. The number of cases usually increase in the countries of Southeast Asia after flood damage during typhoon seasons. This disease also poses a potential threat to Taiwan due to its geographic characteristics. Therefore, it is necessary to establish a practical technique for an estimation of the distribution of Leptospira in natural environment, especially after flood damage. The aim of this study was to demonstrate the use of commercial soil nucleic acid extraction kits with nested PCR in the detection of Leptospira spp. in the farm environment. The detection limit for Leptospira DNA was 24.24 pg. A total of 108 soil samples were collected from farms located within flood-damaged areas; the overall positive rate was 30.6%. Based on the sequence analysis, positive samples were identified as Leptospira interrogans and Leptospira biflexa. This developed approach might be applied in a surveillance of leptospirosis in different countries or in the detection of other pathogens from soils

Disseminated Abscessation Complicated with Bone Marrow Abscess Caused by Arcanobacterium pyogenes in a Goat

An 8-month-old, Nubian wether with a history of systemic illness was euthanatized for a pathological examination. At necropsy, the presence of disseminated abscessation and cellulitis in the limbs was noted. Other postmortem findings associated with the visceral disease in this animal included multiple abscess lesions, mainly in the lungs, kidneys, phalanxes and vertebrae. Histopathological logically, lesions of arteriolitis were found as evidenced by bacterial embolisms in pulmonary and renal arteriola, indicating a bacteremia in the patient. Arcanobacterium pyogenes was consistently isolated from 8 lesions of abscessations, including the lesions of subcutaneous abscesses as well as bone marrow abscess in phalanxes and thoracic vertebrae. This is the first published report of disseminated arcanobacterial infection with bone marrow abscess of both the phalanxes and vertebrae in goat

Synthesis, in vitro macrophage response and detoxification of bamboo charcoal beads for purifying blood

Bamboo charcoal beads (BCBs) were formed by coprecipitating bamboo charcoal particles with chitosan in alkaline solution. The amount of chitosan in the BCBs and their surface properties were measured. When 13-52 mg BCBs were exposed to RAW 264.7 macrophages, the amount of nitric oxide released and the cell viability were close to those of the blank. The amount of cytokine IL-6 secreted by macrophages did not depend on the dose of BCBs but macrophages secreted more TNF-alpha in response to higher doses of BCBs. However, the cytokine levels were relatively low, suggesting the favorable biocompatibility of BCBs. In adsorption experiments, BCBs adsorbed and released bovine serum albumin at particular concentrations, whereas BCBs adsorbed L-phenylalanine without a sign of release. This difference is attributed to the hydrophilicity and the pore size of the BCBs. Finally, the potential of BCBs as biocompatible adsorbents in blood detoxification is considered. (C) 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 94A: 1133-1140, 2010

Characterization of a novel thermophilic, cellulose-degrading bacterium Paenibacillus sp strain B39

Aims: The aims of this study were to identify and characterize the novel thermophilic, cellulose-degrading bacterium Paenibacillus sp. strain B39. Methods and Results: Strain B39 was closely related to Paenibacillus cookii in 16S rRNA gene sequence. Nonetheless, this isolate can be identified as a novel Paenibacillus sp. with respect to its physiological characteristics, biochemical reactions, and profiles of fatty acid compositions. A cellulase with both CMCase and avicelase activities was secreted from strain B39 and purified by ion-exchange chromatography. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, the molecular weight of B39 cellulase was determined as 148 kDa, which was much higher than other cellulases currently reported from Paenibacillus species. The enzyme showed a maximum CMCase activity at 60 degrees C and pH 6.5. Addition of 1 mmol l(-1) of Ca2+ markedly enhanced both CMCase and avicelase activities of the enzyme. Conclusions: We have identified and characterized a novel thermophilic Paenibacillus sp. strain B39 which produced a high-molecular weight cellulase with both CMCase and avicelase activities. Significance and Impact of the Study: Based on the ability to hydrolyse CMC and avicel, the cellulase produced by Paenibacillus sp. strain B39 would have potential applications in cellulose biodegradation