Enzyme Bioprospection of Marine-Derived Actinobacteria from the Chilean Coast and New Insight in the Mechanism of Keratin Degradation in Streptomyces sp. G11C

Marine actinobacteria are viewed as a promising source of enzymes with potential technological applications. They contribute to the turnover of complex biopolymers, such as pectin, lignocellulose, chitin, and keratin, being able to secrete a wide variety of extracellular enzymes. Among these, keratinases are a valuable alternative for recycling keratin-rich waste, which is generated in large quantities by the poultry industry. In this work, we explored the biocatalytic potential of 75 marine-derived actinobacterial strains, focusing mainly on the search for keratinases. A major part of the strains secreted industrially important enzymes, such as proteases, lipases, cellulases, amylases, and keratinases. Among these, we identified two streptomycete strains that presented great potential for recycling keratin wastes—Streptomyces sp. CHA1 and Streptomyces sp. G11C. Substrate concentration, incubation temperature, and, to a lesser extent, inoculum size were found to be important parameters that influenced the production of keratinolytic enzymes in both strains. In addition, proteomic analysis of culture broths from Streptomyces sp. G11C on turkey feathers showed a high abundance and diversity of peptidases, belonging mainly to the serine and metallo-superfamilies. Two proteases from families S08 and M06 were highly expressed. These results contributed to elucidate the mechanism of keratin degradation mediated by streptomycetes.     

Glutamic acid supplementation recovers the reduced performance of weanling pigs fed reduced crude protein diets

This study was conducted to evaluate the supplementation of glutamic acid(Glu)to reduced protein diets on the performance of weanling pigs.One hundred and eighty crossbred weanling pigs([Yorkshire
Landrace]
Duroc,21 d old)having similar body weight(BW)of 6.45 kg were randomly allotted to 1 of 6 dietary treatments(5 pigs per pen[2 barrows and 3 gilts];6 pens per treatment)based on BW and sex during a 6-week trial.Dietary treatments consisted of positive control(PC)diet formulated to have 226.9,205.6,and 188.8 g crude protein(CP)during phases 1,2,and 3,respectively,and negative control(NC)diets with 20 g CP reduction from PC diets and addition of Glu with increasing levels,resulting in the calculated Lys-to-Glu ratios of 1:2.25,1:2.30.1:2.35,1:2.40,and 1:2.45,designated as NC,NC1,NC2,NC3,and NC4,respectively.The BW of pigs receiving PC diet was higher(P<0.05)than those receiving NC diet at d 7,21 and 42.A higher(P<0.05)average daily gain(ADG)from d 1 to 7,8 to 21,22 to 42 and during the overall experiment period was observed in pigs fed PC than NC diet.Pigs fed NC diets including the graded level of Glu linearly increased(P<0.05)BW at d 42,ADG and gain-to-feed ratio(G:F)during the overall experimental period.In addition,trends in linear increase in BW(P=0.056)at d 7 and ADG from d 1 to 7 and d 22 to 42(linear effect,P=0.081,P=0.058 respectively)were observed.A tendency in the linear increment of NH3(P=0.082)at d 21 and linear reduction in methyl mercaptans(P=0.054)emission at d 42 was observed in pigs fed NC diets supplemented with graded level of Glu.In conclusion,supplementing the reduced protein diet with Glu enhanced the growth performance in weanling pigs suggesting that supplementation of Glu can compensate the reduction of 2%CP in the basal diets.     

Effect of an immunomodulator on the efficacy of an attenuated vaccine against avian pneumovirus in turkeys

Since 1997, avian pneumovirus (APV) has caused estimated annual losses of $15 million to the Minnesota turkey industry. In order to develop an attenuated live vaccine against APV, we serially passaged a Minnesota isolate of APV (APV/MN/turkey/1-a/97) in vitro in cell cultures for 41 passages. Laboratory experiments with this high-passage virus (P41) indicated that the attenuated virus provided immunogenic protection to turkeys against challenge with virulent APV, although some birds showed mild to moderate dinical signs after inoculation. To reduce the residual pathogenicity of P41, while maintaining its immunogenicity, we decided to vaccinate turkeys with P41 in the presence of an immunomodulator, S-28828 (1-n-butyl-2-ethoxymethyl-1H-imidazo[4,5-c]quinolin-4-amine-hydrochloride), which is a potent cytokine inducer. The combined inoculation of S-28828 (5 mg/kg body weight) and P41 resulted in a significant reduction in the incidence of virus-induced clinical signs in comparison with birds that received P41 without immunomodulator (P < 0.05). Only 17% of birds inoculated with S-28828 + APV P41 showed mild respiratory symptoms at 5 days postinoculation as compared with 46% of the vaccinated turkeys that did not receive S-28828. Vaccination with either P41 or with P41 + S-28828 protected turkeys against dinical signs and viral replication after challenge with virulent APV. These results indicate that immunomodulators, such as S-28828, may act as good vaccine adjuvants that can reduce the pathogenicity but maintain the immunogenicity of partially attenuated vaccines.     

Serological Methods for Detection of Polymyxa graminis, an Obligate Root Parasite and Vector of Plant Viruses

A purification procedure was developed to separate Polymyxa graminisresting spores from sorghum root materials. The spores were used as im-munogen to produce a polyclonal antiserum. In a direct antigen coating enzyme-linked immunosorbent assay (DAC ELISA), the antiserum could detect one sporosorus per well of the ELISA plate. In spiked root samples, the procedure detected one sporosorus per mg of dried sorghum roots. The majority of isolates of P. graminis from Europe, North America, and India reacted strongly with the antiserum. Interestingly, P. graminis isolates from the state of Rajasthan (northern India), from Pakistan, and an isolate from Senegal (West Africa) reacted weakly with the antiserum. The cross-reactivity of the serum with P. betae isolates from Belgium and Turkey was about 40% of that observed for the homologous isolate. There was no reaction with common fungi infecting roots or with the obligate parasite Olpidium brassicae. However, two isolates of Spongospora sub-terranea gave an absorbance similar to that observed with the homologous antigen. The DAC ELISA procedure was successfully used to detect various stages in the life cycle of P. graminis and to detect infection that occurred under natural and controlled environments. A simple procedure to conjugate antibodies to fluorescein 5-isothiocyanate (FITC) is described. Resting spores could be detected in root sections by using FITC-labeled antibodies. The potential for application of the two serological techniques for studying the epidemiology of peanut clump disease and for the characterization of Polymyxa isolates from various geographical origins is discussed.     

Viewpoints on synergising ASEAN academic visibilities through research collaboration and the establishment of an ASEAN Citation Index Database1

This opinion article expresses two key viewpoints regarding the options for Association of South‐East Asian Nations (ASEAN) universities in pursuit of fulfilling the demand for greater research visibility and academic reputation. The first viewpoint outlines the importance of promoting research collaboration among ASEAN researchers and their international peers in increasing the visibility and impact of ASEAN research. The second viewpoint examines the ASEAN research characteristics, the challenges and existing best practices of its journal publishing landscape, the potential role of non‐ASEAN international journals in improving the profile of ASEAN research and explores the needs and benefits of establishing an ASEAN Integrated Journal Publishing Network (IJPN). It is through these combined approaches, we believe, that effective policy change, the standardisation of publishing structure, procedures, and systems to drive journal development, improvement of journal quality, and establishment of a consistent framework for measuring researcher performance will result. This article also includes a proposal for the IJPN structure and implementation plan, and highlights the key benefits of taking a collaborative approach to achieving this common goal.     

Parental participation in progeny and effective population sizes in experimental seed orchards of wild cherry Prunus avium L. (Batsch)

Parents and progenies genetic diversity, and male and female contributions to the seed crop were assessed in three experimental Prunus avium seed orchards. Collected data were used to compare different effective population sizes, based on phenological, seed crop and paternity analysis. Our results did not show any difference of genetic diversity between parents and progenies. A limited pollen pollution was detected. We showed that distance and coflowering among clones had a significant effect on effective pollination, and a significant effect of the production of flowers was revealed in one of the seed orchards. Our study also revealed a quite low number of effective size of fathers per mother, but high effective sizes of mothers, fathers and parents at the level of the seed orchard. Finally, the calculation of effective size of mothers, fathers and parents was not highly modified when having the complete information based on the paternity analysis.     

Antibacterial activity of embelin

Embelin, a benzoquinone-derivative isolated from Embelia ribes, when tested for its antibacterial potential exhibited significant inhibition against five and moderate activity against three strains of the 12 bacteria tested.     

Insecticidal action of the polyphenolic rich fractions from the stem bark of Streblus asper on Dysdercus cingulatus

Extracts from the stem bark of Streblus asper possess insecticidal activity against the fifth instar of Dysdercus cingulatus. Methanolic extract showed an LC(50) value of 5.56 microg/insect. Partition with chloroform increased the insecticidal activity (LC(50) 2.01 microg/insect). Three polyphenolic rich fractions were obtained from silica-gel column chromatography of the chloroform fraction and found to have noteworthy insecticidal activity (LC(50): 1.82 microg, 2.70 microg, 2.26 microg/insect) by topical application. This may provide a useful beginning for the development of biopesticides.     

Carbon stock and rate of carbon sequestration in Dipterocarpus forests of Manipur,Northeast India

We examined the carbon stock and rate of carbon sequestration in a tropical deciduous forest dominated by Dipterocarpus tuberculatus in Manipur,North East India.Estimation of aboveground biomass was determined by harvest method and multiplied with density of tree species.The aboveground biomass was between18.27–21.922 t ha-1and the carbon stock ranged from9.13 to 10.96 t C ha-1across forest stands.Aboveground biomass and carbon stock increased with the increase in tree girth.The rate of carbon sequestration varied from1.4722 to 4.64136 t ha-1year-1among the dominant tree species in forest stands in tropical deciduous forest area.The rate of carbon sequestration depends on species composition,the density of large trees in different girth classes,and anthropogenic disturbances in the present forest ecosystem.Further work is required to identify tree species having the highest potential to sequester CO2 from the atmosphere,which could lead to recommendations for tree plantations in a degraded ecosystem.