Simple, inexpensive and RNase-free purification of plasmid DNA by fractional precipitation with isopropanol

We present a modified alkaline lysis method for purification of plasmid DNA (pDNA) from bacterial extract using fractional precipitation with isopropanol (FPI). This method includes two successive precipitations with 0.33 and 0.36 volumes of isopropanol and separates pDNA from total RNA and most of the lipopolysaccharides. Using different quality control tests, we demonstrate that plasmids purified with FPI show superior quality compared to plasmids prepared with commercial kits based on spin-column chromatography.

Gut microbiota-based vaccination engages innate immunity to improve blood glucose control in obese mice

Obesity and diabetes increase circulating levels of microbial components derived from the gut microbiota. Individual bacterial factors (i.e., postbiotics) can have opposing effects on metabolic inflammation and blood glucose control. We tested the net effect of gut bacterial extracts on blood glucose using a microbiota-based vaccination strategy in mice. Male and female mice had improved insulin sensitivity and blood glucose control five weeks after a single subcutaneous injection of a specific dose of a bacterial extract obtained from the luminal contents of the proximal gut. Injection of mice with proximal gut extracts from germ-free mice revealed that bacteria were required for a microbiota-based vaccination to improve blood glucose control. Vaccination of Nod1-/-, Nod2-/-, and Ripk2-/- mice showed that each of these innate immune proteins was required for bacterial extract injection to improve blood glucose control. A microbiota-based vaccination promoted a proximal gut immunoglobulin-G (IgG) response directed against bacterial extract antigens, where subcutaneous injection of mice with the luminal contents of the ileum elicited a bacterial extract-specific IgG response that is compartmentalized to the ileum of vaccinated mice. A microbiota-based vaccination was associated with an altered the microbiota composition in the ileum and colon of mice. Lean mice required a single injection of proximal gut bacterial extracts, but high fat diet (HFD)-fed, obese mice required prime-boost bacterial extract injections for improvements in blood glucose control. These data show that, upon subversion of the gut barrier, vaccination with proximal gut bacterial extracts engages innate immunity to promote long-lasting improvements in blood glucose control in a dose-dependent manner.

The natural sequence of events in larval settlement and metamorphosis of Hydroides elegans (Polychaeta; Serpulidae)

The broadly distributed serpulid worm Hydroides elegans has become a model organism for studies of marine biofouling, development and the processes of larval settlement and metamorphosis induced by surface microbial films. Contrasting descriptions of the initial events of these recruitment processes, whether settlement is induced by (1) natural multi-species biofilms, (2) biofilms composed of single bacterial species known to induce settlement, or (3) a bacterial extract stimulated the research described here. We found that settlement induced by natural biofilms or biofilms formed by the bacterium Pseudoalteromonas luteoviolacea is invariably initiated by attachment and secretion of an adherent and larva-enveloping primary tube, followed by loss of motile cilia and ciliated cells and morphogenesis. The bacterial extract containing complex tailocin arrays derived from an assemblage of phage genes incorporated into the bacterial genome appears to induce settlement events by destruction of larval cilia and ciliated cells, followed by attachment and primary-tube formation. Similar destruction occurred when precompetent larvae of H. elegans or larvae of a nudibranch gastropod were exposed to the extract, although neither of them metamorphosed. We argue that larvae that lose their cilia before attachment would be swept away from the sites that stimulated settlement by the turbulent flow characteristic of most marine habitats.

The natural sequence of events in larval settlement and metamorphosis of Hydroides elegans (Polychaeta; Serpulidae)

The broadly distributed serpulid worm Hydroides elegans has become a model organism for studies of marine biofouling, development and the processes of larval settlement and metamorphosis induced by surface microbial films. Contrasting descriptions of the initial events of these recruitment processes, whether settlement is induced by (1) natural multi-species biofilms, (2) biofilms composed of single bacterial species known to induce settlement, or (3) a bacterial extract stimulated the research described here. We found that settlement induced by natural biofilms or biofilms formed by the bacterium Pseudoalteromonas luteoviolacea is invariably initiated by attachment and secretion of an adherent and larva-enveloping primary tube, followed by loss of motile cilia and ciliated cells and morphogenesis. The bacterial extract containing complex tailocin arrays derived from an assemblage of phage genes incorporated into the bacterial genome appears to induce settlement events by destruction of larval cilia and ciliated cells, followed by attachment and primary-tube formation. Similar destruction occurred when precompetent larvae and larvae of a nudibranch gastropod were exposed to the extract, neither of which metamorphosed. We further argue that larvae that lose their cilia before attachment would be swept away from the sites that stimulated settlement by the turbulent flow characteristic of most marine habitats.

Prophylaxis of Upper Airway Infections in a Patient with Partial IgA Deficiency: Concurrent Use of Sublingual Immunotherapy with Inactivated Whole-Cell Bacterial Extract and Der p1

Selective IgA deficiency is the most common type of primary immunodeficiency, but there is not yet a specific effective treatment. The most prevalent clinical manifestations are infectious diseases of the respiratory system. We report herein the case of an 11-year-old female with selective IgA deficiency and recurring episodes of respiratory infections associated with rhinitis and asthma. We evaluated the efficacy of sublingual immunotherapy combined with inactivated whole-cell bacterial extract and Der p1-specific immunotherapy. After 18 months of clinical follow-up, we observed a significant reduction in the number of episodes of respiratory infections associated with control of atopic diseases. We also observed a 3-fold increase in serum IgA levels compared to treatment initiation. This case demonstrates the potential utility of the concurrent use of sublingual immunotherapy with inactivated whole-cell bacterial extract and Der p1 for successful control of allergy and infection in partial selective IgA deficiency.

AKTIVITAS SENYAWA ANTIBAKTERI DAN ANTI-UV DARI Phyllidia varicosa (Cuvier, 1804) DAN BAKTERI SIMBIONNYA (NUDIBRANCHIA GASTROPODA) DARI PERAIRAN TANJUNG MANDOLANG, MINAHASA

Phyllidia varicosa is an organism belongs to the order Nudibranchia, which is commonly known as a sea rabbit. This organism is able to synthesize secondary metabolites from food. The purpose of this study was to obtain P. varicosa extract and symbiotic bacterial extract from P. varicosa, then determine the antibacterial activity of P. varicosa extract and the symbionic bacteria extract against Escherichia coli DSM498, Bacillus megaterium DSM32T and anti-UV activity. P. varicose symbiotic bacterial were isolated and extracted. The results obtained 5 bacterial isolates. The results of antibacterial assay of isolates PhVa 1.1, PhVa 1.3, PhVa 2.1, PhVa 2.3 and PhVa 2.4 shown that these isolates have an antibacterial activity against E. coli DSM498 and B. megaterium DSM32T. Anti-UV assay results shown an absorption at UV-A with the highest value of 1.991 at λ 340 nm. Keywords: Nudibranchia, Phyllidia varicosa, Antibacterial, Anti-UV, Escherichia coli, Bacillus megaterium Abstrak Phyllidia varicosa merupakan organisme yang termasuk dalam ordo Nudibranchia, yang umumnya dikenal sebagai kelinci laut. Organisme ini mampu mensintesis metabolit sekunder dari bahan makanannya. Tujuan dari penelitian ini yaitu untuk mendapatkan ekstrak P. varicosa dan ekstrak bakteri simbion dari P. varicosa, kemudian mengamati aktivitas antibakteri dari ekstrak P. varicosa dan ekstrak bakteri simbionnya terhadap bakteri Escherichia coli DSM498 dan Bacillus megaterium DSM32T serta menguji aktivitas anti-UV. Bakteri yang bersimbion dengan P. varicosa diisolasi dan diekstraksi, lalu diuji bioaktivitas antibakteri dan diuji anti-UV terhadap ekstrak P. varicosa dan ekstrak bakteri simbionnya. Hasil akhir dari penelitian ini yaitu didapatkan 5 isolat bakteri. Hasil uji aktivitas antibakteri yaitu isolat PhVa 1.1, PhVa 1.3, PhVa 2.1, PhVa 2.3 dan PhVa 2.4 memiliki aktivitas antibakteri terhadap bakteri E. coli DSM498 dan B. megaterium DSM32T. Hasil uji anti-UV menunjukkan serapan pada UV-A dengan nilai tertinggi 1,991 pada λ 340 nm. Kata kunci: Nudibranchia, Phyllidia varicosa, Antibakteri, Anti-UV, Escherichia coli, Bacillus megaterium

Isolation of bacteria from Apis cerana hive, their antibacterial potency and cytotoxicity

Yasmin Y, Fitri L, Fauziah, Wasliyah F. 2019. Isolation of bacteria from Apis cerana hive, their antibacterial potency and cytotoxicity. Biodiversitas 20: 2733-2738. This study aimed to identify bacteria from Apis cerana hive; to determine their antibacterial activity, and cytotoxic effect of hive extract as well as bacterial extract. Identification of bacteria isolated from the hive was carried out based on morphological and biochemical characters. The antibacterial assay of beehive bacteria isolates was done by disk diffusion method against Staphylococcus aureus and Escherichia coli. Toxicity test of beehive extract and beehive bacterial extract were carried out using Brine Shrimp Lethality Test (BSLT) with six final concentrations (32 ppm, 64 ppm, 126 ppm, 250 ppm, 500 ppm, and 1000 ppm). The LC50 value was determined by probit analysis using SPSS. Bacterial isolation showed there were four bacterial isolates from A. cerana hive were the genus Bacillus (BSL1, and BSL3), the genus Micrococcus (BSL2) and the genus Neisseria (BSL4). The antibacterial assay showed that BSL4 isolate has the highest antibacterial activity against S. aureus with the diameter of the inhibitory zone was 32.6 mm. Toxicity test showed that A. cerana hive extract and bacterial ethanol extract from A. cerana hive had LC50 value of 67,744 ppm and 86.985 ppm respectively and categorized as toxic.

Phytochemical analysis, proximate composition and antibacterial activities of Ziziphus Species (Z. jujube and Z. spina christi)

Despite tremendous progress in human medicines infectious diseases caused by bacteria, fungi, viruses and parasites are still a major threat to public health. Their impact is particularly large in developing countries due to relative unavailability of medicines and emergence of widespread drugs resistance (Zampini 2009).The aim and objectives of this research work was designed to carried out the Phytochemical analysis, proximate composition and evaluate the antimicrobial activities of the Ziziphus jujube and Ziziphus spina christi leaves against clinical bacterial isolates (Escherichia coli, Staphloccoccus aureus and Klebsiella pneumoniae) as they were found out that since ancient times to date, they are used in treating various antimicrobial, ailmentand disorders etc. The results of the research shows that, the effect of antibacterial activities of both aqueous and ethanolic extracts of Ziziphus jujube (e extract conc. aqueous; F=119.37, Bacterial extract conc. aqueous; F=1.00 and extract conc. ethanol; F=15.74, Bacterial extract conc. ethanol; F=0.59) are reciprocal proportional to their counterpart, Ziziphus spina-christi (extract conc. aqueous; F=54.96, Bacterial extract conc. aqueous; F=0.94 and extract conc. ethanol; F=81.11, Bacterial extract conc. ethanol; F=1.37). In sum, the minimum inhibitory concentration of Ziziphus jujube shows that, the aqueous extract has M.I.C at range of 11.7 to 8.7mg/ml on all tested bacteria but the ethanolic extract has M.I.C of 14.8 to 8.2mg/ml range on E.coli, Klepsiella spp and S. aureus. While, the minimum inhibitory concentraton of Ziziphus spina-christi shows that the M.I.C of aqueous extract range of 12.8 to 8.3mg/ml on E. coli, Klepsiella spp and S.aureus. But, Ziziphus spina-christi M.I.C of ethanolic extract is 13.5 to 8.8mg/ml on all the tested bacteria. In sum, Zizuphus spina-christi has lower nutritional content and low MIC ethanolic extract than that of Ziziphus jujube.