Green tree frog

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Identification and bioactivity evaluation of two novel temporins from the skin secretion of the European edible frog, Pelophylax kl. Esculentus

The amphibian temporins, amongst the smallest antimicrobial peptides (AMPs), are α-helical, amphipathic, hydrophobic and cationic and are active mainly against Gram-positive bacteria but inactive or weakly active against Gram-negative bacteria. Here, we report two novel members of the temporin family, named temporin-1Ee (FLPVIAGVLSKLFamide) and temporin-1Re (FLPGLLAGLLamide), whose biosynthetic precursor structures were deduced from clones obtained from skin secretion-derived cDNA libraries of the European edible frog, Pelophylax kl. esculentus, by ‘shotgun’ cloning. Deduction of the molecular masses of each mature processed peptide from respective cloned cDNAs was used to locate respective molecules in reverse-phase HPLC fractions of secretion. Temporin-1Ee (MIC = 10 μM) and temporin-1Re (MIC = 60 μM) were both found to be active against Gram-positive Staphylococcus aureus, but retaining a weak haemolytic activity. To our knowledge, Single-site substitutions can dramatically change the spectrum of activity of a given temporin. Compared with temporine-1Ec, just one chemically-conservative substitution (Val8 instead of Leu8), temporin-1Ee bearing a net charge of +2 displays broad-spectrum activity with particularly high potency on the clinically relevant Gram-negative strains, Escherichia coli (MIC = 40 μM). These factors bode well for translating temporins to be potential drug candidates for the design of new and valuable anti-infective agents.

Peptidomic approach identifies cruzioseptins, a new family of potent antimicrobial peptides in the splendid leaf frog, Cruziohyla calcarifer

Phyllomedusine frogs are an extraordinary source of biologically active peptides. At least 8 families of antimicrobial peptides have been reported in this frog clade, the dermaseptins being the most diverse. By a peptidomic approach, integrating molecular cloning, Edman degradation sequencing and tandem mass spectrometry, a new family of antimicrobial peptides has been identified in Cruziohyla calcarifer. These 15 novel antimicrobial peptides of 20–32 residues in length are named cruzioseptins. They are characterized by having a unique shared N-terminal sequence GFLD– and the sequence motifs –VALGAVSK– or –GKAAL(N/G/S) (V/A)V– in the middle of the peptide. Cruzioseptins have a broad spectrum of antimicrobial activity and low haemolytic effect. The most potent cruzioseptin was CZS-1 that had a MIC of 3.77 μM against the Gram positive bacterium, Staphylococcus aureus and the yeast Candida albicans. In contrast, CZS-1 was 3–fold less potent against the Gram negative bacterium, Escherichia coli (MIC 15.11 μM). CZS-1 reached 100% haemolysis at 120.87 μM. Skin secretions from unexplored species such as C. calcarifer continue to demonstrate the enormous molecular diversity hidden in the amphibian skin. Some of these novel peptides may provide lead structures for the development of a new class of antibiotics and antifungals of therapeutic use.

Baltikinin: A New Myotropic Tryptophyllin-3 Peptide Isolated from the Skin Secretion of the Purple-Sided Leaf Frog, Phyllomedusa baltea

Here we report the identification of a novel tryptophyllin-3 peptide with arterial smooth muscle relaxation activity from the skin secretion of the purple-sided leaf frog, Phyllomedusa baltea. This new peptide was named baltikinin and had the following primary structure, pGluDKPFGPPPIYPV, as determined by tandem mass spectrometry (MS/MS) fragmentation sequencing and from cloned skin precursor-encoding cDNA. A synthetic replicate of baltikinin was found to have a similar potency to bradykinin in relaxing arterial smooth muscle (half maximal effective concentration (EC50) is 7.2 nM). These data illustrate how amphibian skin secretions can continue to provide novel potent peptides that act through functional targets in mammalian tissues.

Identification of Miscellaneous Peptides from the Skin Secretion of the European Edible Frog, Pelophylax kl. Esculentus

The chemical compounds synthesised and secreted from the dermal glands of amphibian have diverse bioactivities that play key roles in the hosts' innate immune system and in causing diverse pharmacological effects in predators that may ingest the defensive skin secretions. As new biotechnological methods have developed, increasing numbers of novel peptides with novel activities have been discovered from this source of natural compounds. In this study, a number of defensive skin secretion peptide sequences were obtained from the European edible frog, P. kl. esculentus, using a 'shotgun' cloning technique developed previously within our laboratory. Some of these sequences have been previously reported but had either obtained from other species or were isolated using different methods. Two new skin peptides are described here for the first time. Esculentin-2c and Brevinin-2Tbe belong to the Esculentin-2 and Brevinin-2 families, respectively, and both are very similar to their respective analogues but with a few amino acid differences. Further, [Asn-3, Lys-6, Phe-13] 3-14-bombesin isolated previously from the skin of the marsh frog, Rana ridibunda, was identified here in the skin of P. kl. esculentus. Studies such as this can provide a rapid elucidation of peptide and corresponding DNA sequences from unstudied species of frogs and can rapidly provide a basis for related scientific studies such as those involved in systematic or the evolution of a large diverse gene family and usage by biomedical researchers as a source of potential novel drug leads or pharmacological agents.

Ex Vivo Smooth Muscle Pharmacological Effects of a Novel Bradykinin-Related Peptide, and Its Analogue, from Chinese Large Odorous Frog, Odorrana livida Skin Secretions

Bradykinin-related peptides (BRPs) are one of the most extensively studied frog secretions-derived peptide families identified from many amphibian species. The diverse primary structures of BRPs have been proven essential for providing valuable information in understanding basic mechanisms associated with drug modification. Here, we isolated, identified and characterized a dodeca-BRP (RAP-L1, T6-BK), with primary structure RAPLPPGFTPFR, from the skin secretions of Chinese large odorous frogs, Odorrana livida. This novel peptide exhibited a dose-dependent contractile property on rat bladder and rat ileum, and increased the contraction frequency on rat uterus ex vivo smooth muscle preparations; it also showed vasorelaxant activity on rat tail artery smooth muscle. In addition, the analogue RAP-L1, T6, L8-BK completely abolished these effects on selected rat smooth muscle tissues, whilst it showed inhibition effect on bradykinin-induced rat tail artery relaxation. By using canonical antagonist for bradykinin B1 or B2 type receptors, we found that RAP-L1, T6-BK -induced relaxation of the arterial smooth muscle was very likely to be modulated by B2 receptors. The analogue RAP-L1, T6, L8-BK further enhanced the bradykinin inhibitory activity only under the condition of co-administration with HOE140 on rat tail artery, suggesting a synergistic inhibition mechanism by which targeting B2 type receptors.

Improving income and livelihood of poor farming household in Bangladesh through adoption of improved aquaculture technologies and varieties

Fish are an important part of Bangladeshi culture and diet. Bangladesh ranks among the top five freshwater fish producers in the world. Fish are abundant in the thousands of rivers, ponds, lakes and seasonal floodplains across the country. They are a major source of protein for people living near these water bodies. In Bangladesh, many households depend on fish farming for their livelihood. By growing fish in homestead ponds, households have a consistent supply of nutritious fish and can sell the surplus for an income. The USAID-funded Cereal Systems Initiative for South Asia in Bangladesh (CSISA-BD) aimed to increase the income of farming households through increased productivity of aquaculture systems. Key activities of the project included developing and disseminating appropriate improved agricultural technology and quality fish seeds to improve livelihoods, food security and nutrition.

Delivering Regional Extension in Qld Farming Systems - Central Queensland

The CQ Cotton Regional Extension project has been a key to the delivery of emerging, cutting edge research information and knowledge to the Central Queensland cotton industry. The direct relevance of southern research to cotton production under the conditions experienced in CQ always has been an issue which could be addressed through regional assessment and adaptation. The project links the national research to the region through development and extension, with a strong focus on the major industry production issues including but not limited to disease, Integrated Pest Management (IPM), soils, nutrition and integrated weed management. Susan Mass has supported the implementation of national industry-wide programs particularly the industry Best Management Practices program (myBMP). This project has successfully transitioned to a focus on delivering national outcomes in target lead areas as part of National Development and Delivery Team established by Cotton CRC, CRDC and Cotton Australia, while maintaining a regional extension presence for Central Queensland cotton & grain farming systems. Susan Mass has very effectively merged and integrated strong regional extension support to cotton growers in Central Queensland with delivery of industry extension priorities across the entire industry in the Development and Delivery Team model. Susan is the target lead for disease and farm hygiene. Recognising the challenges of having regionally relevant research in Central Queensland, this project has facilitated locally based research including boll rot, Bt cotton resistance management, and mealybug biology through strong collaborations. This collaborative approach has included linkage to Department of Environment and Resource Managmeent (DERM) groups and myBMP programs resulting in a high uptake in CQ.

Strategic tillage in no-till farming systems in Australia's northern grains-growing regions: I. Drivers and implementation

Development of no-tillage (NT) farming has revolutionized agricultural systems by allowing growers to manage greater areas of land with reduced energy, labour and machinery inputs to control erosion, improve soil health and reduce greenhouse gas emission. However, NT farming systems have resulted in a build-up of herbicide-resistant weeds, an increased incidence of soil- and stubble-borne diseases and enrichment of nutrients and carbon near the soil surface. Consequently, there is an increased interest in the use of an occasional tillage (termed strategic tillage, ST) to address such emerging constraints in otherwise-NT farming systems. Decisions around ST uses will depend upon the specific issues present on the individual field or farm, and profitability and effectiveness of available options for management. This paper explores some of the issues with the implementation of ST in NT farming systems. The impact of contrasting soil properties, the timing of the tillage and the prevailing climate exert a strong influence on the success of ST. Decisions around timing of tillage are very complex and depend on the interactions between soil water content and the purpose for which the ST is intended. The soil needs to be at the right water content before executing any tillage, while the objective of the ST will influence the frequency and type of tillage implement used. The use of ST in long-term NT systems will depend on factors associated with system costs and profitability, soil health and environmental impacts. For many farmers maintaining farm profitability is a priority, so economic considerations are likely to be a primary factor dictating adoption. However, impacts on soil health and environment, especially the risk of erosion and the loss of soil carbon, will also influence a grower's choice to adopt ST, as will the impact on soil moisture reserves in rainfed cropping systems. © 2015 Elsevier B.V.

A manual for improving fish production in Northern Zambia through integrated farming systems

This manual was written as part of the Integrated Research in Development for Improved Livelihoods Programme in Northern Province, Zambia (IRDLP) and is primarily intended for extension agents to use with smallholder farmers engaged in semi-intensive fish farming in Northern Zambia. The IRDLP is an Irish Aid-funded project implemented by WorldFish, Harvest Plus and the Center for International Forestry Research (CIFOR). The goal of the IRDLP is to help improve the livelihoods, health status, and food and nutrition security of resource-poor households in the Mbala and Luwingu districts in Northern Zambia, especially women and vulnerable groups. This is achieved through generating and providing evidence-based information, scientific technologies and livelihood solutions to trigger community and farmer innovations for positive change. This manual provides information on how smallholder fish farmers can improve fish production in Northern Zambia, particularly in the Luwingu and Mbala districts, through integrated farming practices.

EFFECT OF FARMING PRACTICES AND FARM HISTORY ON INCIDENCE OF COCONUT LETHAL YELLOWING IN MOZAMBIQUE

Management of coconut ( Cocos nucifera ) lethal yellowing disease (CLYD), which has killed about eight million coconut trees in Mozambique, has proved challenging. The objective of this study was to investigate the impact of farming practices and related history, on the CLYD incidence in Mozambique. The methodology included a socioeconomic questionnaire to the households and direct observations on the palm farms. The collected data were analysed using logistic regression. Five out of 11 explanatory variables tested, namely farm age, availability of other palm species on the coconut farm, type of coconut varieties grown, root cut practices, and intercropping had a significant (P< 0.05) effect on CLYD incidence. Coconut farms <10 years had higher odds of higher disease incidence compared to the farms between 10 to 40 years old. The presence of other palm species in the coconut farms had two times higher odds of having higher disease incidence levels compared to farms without other palm species. Tall coconut varieties were likely to be more tolerant to CLYD compared to dwarf varieties. Coconut farms with some kind of intercropping had two times higher odds of having higher disease incidence levels compared to pure stands. The practice of cutting coconut roots had three times higher odds of having high disease incidence levels compared to non-practicing farms. Farm age, availability of other palm species on the coconut farm, type of coconut varieties grown, root cut practices and intercropping need to be considered for integrated CLYD management.

Stimulation of bacterial exoproteolytic activity by fish farming in coastal marine ponds: Effect on dissolved protein cycling

The effect of fish farming on dissolved amino acid concentrations, bacterioplankton abundance and exoproteolytic activity was assessed in 3 experimental marine ponds. Different standing stocks of fish were introduced (semi-intensive pond: 250 g.m(-2); semi-extensive pond: 50 g.m(-2) control pond: 0). Sea bass farming increased dissolved combined amino acid (DCAA) concentrations only in the semi-intensive pond. Bacterial standing stock was unaffected by fish food supply. However, bacterial exoproteolytic activity was strongly stimulated by aquaculture intensification; the average maximal rate of dissolved protein hydrolysis (V-m) increased with intensity (control pond: 1 500 nM.h(-1); semi-extensive pond: 2 600 nM.h(-1) semi-intensive pond: 5 100 nM.h(-1)). DCAA fluxes through bacterial exoproteolytic activity ranged between 16 (semi-extensive) and 11% (semi-intensive) of the daily nitrogen input by fish food. Bacterial exoproteolytic activity allowed a substantial part of the increased supply of dissolved amino nitrogen to be incorporated into bacterial biomass, then available for transfer to higher trophic levels within the ponds. It also significantly decreased dissolved organic nitrogen export from the ponds to the surrounding environment.