Piper viridescens sp. nov. (Piperaceae) from Thailand

Piper viridescens sp. nov. is proposed based on material from northern Thailand. The new species is described and illustrated, resulting from a revision of the genus in Thailand. The new species is most closely related to P. boehmeriifolium (Miq.) C. DC. A table comparing diagnostic features of P. viridescens, P. boehmeriifolium and P. retrofractum is provided.     

Mosquito larvicidal activity of polyacetylenes from species in the Asteraceae

Twenty-four polyacetylenes isolated from species in the Asteraceae were screened for their near-UV-mediated larvicidal properties to the mosquito, Aedes aegypti. Dosage resonse lines for three of the most active compounds were prepared. One of these, σ-terthienyl from Tagetes was more toxic than DDT when near-UV radiation was provided. A scaled up trial with this compound in a simulated field situation demonstrated its potential as a larvicide. The effects of the radiation dose on larval survival were studied and the action spectra for the phototoxic effects of three compounds on larvae were obtained with narrowband interference filters. Because of the close agreement between the action and absorption spectra, it appears most probable that the polyacetylene is the primary absorbing compound. The potential of polyacetylenes as larvicides and their role in plant herbivore relations are discussed.     

Mosquito larvicidal activity of Escherichia coli with combinations of genes from Bacillus thuringiensis subsp. israelensis.

The genes cryIVA and cryIVD, encoding 134- and 72-kDa proteins, respectively, and the gene for a regulatory 20-kDa polypeptide of Bacillus thuringiensis subsp. israelensis (serovar H14) were cloned in all seven possible combinations by the Escherichia coli expression vectors pT7 and pUHE. The four combinations containing cryIVA (cryIVA alone, with cryIVD, with the 20-kDa-protein gene, and with both) displayed high levels of mosquito larvicidal activity in pUHE. The toxicity of the combination of cryIVA and cryIVD, with or without the 20-kDa-protein gene, was higher than has ever been achieved with delta-endotoxin genes in recombinant E. coli. Fifty percent lethal concentrations against third-instar Aedes aegypti larvae for these clones decreased (i.e., toxicity increased) continuously to about 3 x 10(5) cells ml-1 after 4 h of induction. Larvicidal activities, obtained after 30 min of induction, were lower for clones in pT7 and decreased for an additional 3.5 h. Induction of either cryIVD or the 20-kDa-protein gene alone resulted in no larvicidal activity in either pT7 or pUHE20. Cloned together, these genes were slightly toxic in pT7 but not in pUHE20. Five minutes of induction of this combination (cryIVD with the 20-kDa-protein gene) in pT7 yielded a maximal mortality of about 40%, which decreased rapidly and disappeared completely after 50 min. CryIVD is thus apparently degraded in E. coli and partially stabilized by the 20-kDa regulatory protein. Larvicidal activity of the combination of cryIVA and cryIVD was sevenfold higher than that of cryIVA alone, probably because of the cross-stabilization of the polypeptides or the synergism between their activities.     

Mosquito larvicidal activity of transgenic Anabaena PCC 7120 expressing toxin genes from Bacillus thuringiensis subsp. israelensis

Genes encoding the mosquito larvicidal toxins Cry4Aa, Cry11Aa, Cyt1Aa and the regulatory P20 from Bacillus thuringiensis subsp. israelensis were introduced into the nitrogen-fixing, filamentous cyanobacterium Anabaena PCC 7120 for expression under control of two strong promoters P(psbA) and P(A1). The clone pRVE4-ADRC displayed toxicity against fourth-instar larvae of Aedes aegypti, the highest ever achieved in cyanobacteria. It was about 2.5-fold more toxic than the respective clone without cyt1Aa [Wu et al., Appl. Environ. Microbiol. 63 (1997) 4971-4975]. Cyt1Aa synergized the combination of Crys by about five-fold. Consistently, the lethal times exerted by pRVE4-ADRC were also reduced (it killed exposed larvae more quickly). This clone may become a useful biological control agent which reduces the probability of resistance development in the target organisms [Wirth et al., Proc. Natl. Acad. Sci. USA 94 (1997) 10536-10540].     

Rapid identification of bacterial isolates from aqueous kava (Piper methysticum) extracts by polymerase chain reaction and DNA sequencing

Aim:  Fresh kava beverages have a limited shelf life under refrigerated conditions. The objective of this study was to isolate and identify bacteria in aqueous extracts of kava rhizome.
Methods and Results:  The internal part of kava rhizome was used to minimize soil contamination. Three kava extracts were prepared, serially diluted and plated on nutrient agar. Isolated colonies were identified by sequencing polymerase chain reaction amplicons targeting the eubacterial 16S rDNA and the tuf gene of Staphylococcus . Seventy-five bacterial isolates belonged to 16 genera. Bacillus , Cellulomonas , Enterococcus , Pectobacterium and Staphylococcus were identified in all kava extracts.
Conclusions:  Kava rhizome contains large amounts of starch and fibre, which justify the presence of polysaccharide-degrading bacteria in the extracts. Bacillus cereus group and Staphylococcus species may produce toxins and cause foodborne illness.
Significance and Impact of the Study:  The results of this study provide fundamental information that may be used to enhance the microbial quality and safety of kava beverages.     

Mosquito larvicidal activity of transgenic Anabaena strain PCC 7120 expressing combinations of genes from Bacillus thuringiensis subsp. israelensis.

Various combinations of the genes cryIVA (cry4A), cryIVD (cry11A), and p20 from Bacillus thuringiensis subsp. israelensis were introduced into the nitrogen-fixing cyanobacterium Anabaena sp. strain PCC 7120 by means of Escherichia coli-Anabaena shuttle vector pRL488p and were expressed under control of two tandem strong promoters, a cyanobacterial promoter (PpsbA) and an E. coli T7 promoter (PA1). Two of the clones carrying cryIVA plus cryIVD, one with p20 and one without p20, displayed toxicity against third-instar larvae of Aedes aegypti at levels greater than any level previously reported for transgenic cyanobacteria.     

Comparison of the antioxidant activity of aqueous and ethanolic extracts from chaga (Inonotus obliquus)

Additional extraction of biologically active compounds from a chaga shred has been carried out after completion of the aqueous extraction. The extraction was carried out using ethanol. We have shown that the antioxidant activity of the ethanolic extracts and their components is higher than that of the aqueous extracts.     

Chemical composition,oviposition deterrent and larvicidal activities against Aedes aegypti of essential oils from Piper marginatum Jacq. (Piperaceae)

The essential oils of leaves, stems and inflorescences of Piper marginatum, harvested in the Atlantic forest in the State of Pernambuco, Brazil, were obtained by hydrodistillation. GC and GC–MS analyses revealed the presence of 40 components accounting, respectively, for 99.6%, 99.7% and 99.1% of the leaf, stem and inflorescence oil, the most abundant being (Z)- or (E)-asarone and patchouli alcohol. The essential oil of the inflorescences exhibited potent activity against the 4th instar of Aedes aegypti with LC₁₀ and LC₅₀ values of 13.8 and 20.0 ppm, respectively. Furthermore, the inflorescence oil did not interfere in the oviposition of A. aegypti females when assayed at 50 ppm. These properties suggest that P. marginatum oil is a potential source of valuable larvicidal compounds for direct use or in conjunction with baits in traps constructed to capture eggs and larvae.     

Antibacterial activity of extracts and neolignans from Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck

The evaluation of the activity of the aqueous and ethyl acetate extracts of the leaves of Piper regnellii was tested against gram-positive and gram-negative bacteria. The aqueous extract displayed a weak activity against Staphylococcus aureus and Bacillus subtilis with minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of 1000 micrograms/ml. The ethyl acetate extract presented a good activity against S. aureus and B. subtilis with MIC and MBC at 15.62 micrograms/ml. In contrast to the relative low MICs for gram-positive bacteria, gram-negative bacteria were not inhibited by the extracts at concentrations < or = 1000 mg/ml. The ethyl acetate extract was fractionated on silica gel into nine fractions. The hexane and chloroform fractions were active against S. aureus (MIC at 3.9 micrograms/ml) and B. subtilis (MIC at 3.9 and 7.8 micrograms/ml, respectively). Using bioactivity-directed fractionation, the hexane fraction was rechromatographed to yield the antimicrobial compounds 1, 2, 5, and 6 identified as eupomatenoid-6, eupomatenoid-5, eupomatenoid-3, and conocarpan, respectively. The pure compounds 1 and 2 showed a good activity against S. aureus with MIC of 1.56 micrograms/ml and 3.12 micrograms/ml, respectively. Both compounds presented MIC of 3.12 micrograms/ml against B. subtilis. The pure compound 6 named as conocarpan was quite active against S. aureus and B. subtilis with MIC of 6.25 micrograms/ml. The antibacterial properties of P. regnellii justify its use in traditional medicine for the treatment of wounds, contaminated through bacteria infections.     

A study of the larvicidal activity of Origanum (Labiatae) species from southwest Turkey.

The insecticidal activity of essential oils isolated from two species of Origanum (Origanum onites L. and Origanum minutiflorum (O. Schwarz and P.H. Davis) (Labiate)) was investigated against 3rd and 4th instar larvae of the mosquito, Culex pipiens L. (Diptera: Culicidae). The LC50 and LC90 values were 24.8 and 61.3 ppm, and 73.8 and 118.9 ppm, respectively. This activity was attributed to the carvacrol, the major constituent of these oils.     

Insecticidal activity of Piper tuberculatum Jacq. extracts: synergistic interaction of piperamides

Abstract

• 1 The insecticidal activity of the neotropical pepper Piper tuberculatum Jacq. and its isolated piperamides was studied. Bioassays with the mosquito Aedes atropalpus L. assessed the relative toxicity of the whole extract of Piper tuberculatum and four of the piperamides, which were isolated and identified, then prepared synthetically.
• 2 The results confirm that P. tuberculatum leaf extracts are as effective as black pepper seed extract and provide an alternative pepper insecticide from a more convenient source, the leaves.
• 3 Experiments with piperamides showed that the tertiary and quaternary mixtures have greater‐than‐additive toxicity compared to single compounds or binary mixtures. One of the four amide compounds, 4,5‐dihydropiperlonguminine, was the most acutely toxic in mosquito larvae bioassays.
• 4 A study of piperamide levels from different P. tuberculatum populations in Costa Rica determined that they were relatively homogeneous. Piper tuberculatum from only one of the five sites had higher levels of one piperamide, 4,5‐dihydropiperine, in both leaf and stem parts. One explanation for differences in the amide concentrations between populations is that one site is ecologically unique compared to the other four.

     

Piperidine alkaloids from Piper retrofractum Vahl. protect against high-fat diet-induced obesity by regulating lipid metabolism and activating AMP-activated protein kinase

The fruits of Piperretrofractum Vahl. have been used for their anti-flatulent, expectorant, antitussive, antifungal, and appetizing properties in traditional medicine, and they are reported to possess gastroprotective and cholesterol-lowering properties. However, their anti-obesity activity remains unexplored. The present study was conducted to isolate the anti-obesity constituents from P. retrofractum Vahl. and evaluate their effects in high-fat diet (HFD)-induced obese mice. Piperidine alkaloids from P. retrofractum Vahl. (PRPAs), including piperine, pipernonaline, and dehydropipernonaline, were isolated as the anti-obesity constituents through a peroxisome proliferator-activated receptor δ (PPARδ) transactivation assay. The molecular mechanism was investigated in 3T3-L1 adipocytes and L6 myocytes. PRPA treatment activated AMP-activated protein kinase (AMPK) signaling and PPARδ protein and also regulated the expression of lipid metabolism-related proteins. In the animal model, oral PRPA administration (50, 100, or 300 mg/kg/day for 8 weeks) significantly reduced HFD-induced body weight gain without altering the amount of food intake. Fat pad mass was reduced in the PRPA treatment groups, as evidenced by reduced adipocyte size. In addition, elevated serum levels of total cholesterol, low-density lipoprotein cholesterol, total lipid, leptin, and lipase were suppressed by PRPA treatment. PRPA also protected against the development of nonalcoholic fatty liver by decreasing hepatic triglyceride accumulation. Consistent with the in vitro results, PRPA activated AMPK signaling and altered the expression of lipid metabolism-related proteins in liver and skeletal muscle. Taken together, these findings demonstrate that PRPAs attenuate HFD-induced obesity by activating AMPK and PPARδ, and regulate lipid metabolism, suggesting their potential anti-obesity effects.     

Mosquito larvicidal activity of Aloe vera (Family: Liliaceae) leaf extract and Bacillus sphaericus,against Chikungunya vector,Aedes aegypti

The bio-efficacy of Aloe vera leaf extract and bacterial insecticide, Bacillus sphaericus larvicidal activity was assessed against the first to fourth instars larvae of Aedes aegypti, under the laboratory conditions. The plant material was shade dried at room temperature and powdered coarsely. A. vera and B. sphaericus show varied degrees of larvicidal activity against various instars larvae of A. aegypti. The LC₅₀ of A. vera against the first to fourth instars larvae were 162.74, 201.43, 253.30 and 300.05 ppm and the LC₉₀ 442.98, 518.86, 563.18 and 612.96 ppm, respectively. B. sphaericus against the first to fourth instars larvae the LC₅₀ values were 68.21, 79.13, 93.48, and 107.05 ppm and the LC₉₀ values 149.15, 164.67, 183.84, and 201.09 ppm, respectively. However, the combined treatment of A. vera + B. sphaericus (1:2) material shows highest larvicidal activity of the LC₅₀ values 54.80, 63.11, 74.66 and 95.10 ppm; The LC₉₀ values of 145.29, 160.14, 179.74 and 209.98 ppm, against A. aegypti in all the tested concentrations than the individuals and clearly established that there is a substantial amount of synergist act. The present investigation clearly exhibits that both A. vera and B. sphaericus materials could serve as a potential larvicidal agent. Since, A. aegypti is a container breeder vector mosquito this user and eco-friendly and low-cost vector control strategy could be a viable solution to the existing dengue disease burden. Therefore, this study provides first report on the mosquito larvicidal activity the combined effect of A. vera leaf extract and B. sphaericus against as target species of A. aegypti.     

Efficacy of botanical insecticides from Piper species (Piperaceae) extracts for control of Ruropean chafer (Coleoptera: Scarabaeidae)

Biopesticides, including botanicals, can offer a safe and effective alternative to conventional insecticides for controlling major insect pests within an integrated pest management program. The current study highlights the practical application of a botanical insecticide for controlling a major insect pest of turfgrass: European chafer, Rhizotrogus majalis (Razoumowsky). Greenhouse and field trials were conducted to test the efficacy of a botanical formulation based on black pepper, Piper nigrum L. (Piperaceae), seed extracts to R. majalis larvae. The 7-d P. nigrum extract LC50 for R. majalis third instars was 2.5%. Successful treatment in the field was accomplished with the application of a 2% P. nigrum formulation to turfgrass infested with R. majalis second and third instars, whereas 4% extract was required in a second field trial with older third instars. The 2% pepper extract activity was comparable with the conventional insecticide diazinon in the first field trial. However, the 4% pepper extracts significantly affected the earthworm populations in treated plots compared with diazinon in the second field trail. The analysis of soil residues for piperamides in the P. nigrum extract determined a half-life of 1 - 2.6 d in the first and second field trials, respectively. This confirmed the expectation that under field conditions the residual activity would be less than conventional insecticides, thereby reducing the environmental risk associated with pesticide use. We recommend the pepper formulation for spot treatment applications when population densities reveal an epicenter of infestation rather than broadcasting over large areas, thus helping to minimize cost and negative affects on nontarget invertebrates.     

Prenylhydroxybenzoic acid compounds with pungent activity from Piper arieianum (CDC) leaves

Prenylhydroxybenzoic acid derivatives and other two analogues previously reported were isolated from Piper arieianum leaves. The structures of the compounds were assigned from detailed spectroscopical analysis (NMR 1D and 2D and HR ESI TOF MS) data and by comparison with data from the literature. These molecules posses pungent activity different to that of capsaicin and their activity is related to their structure and their mechanism of action can involve interactions with TRPV1 channel.