Candidate Biocontrol Agent Research Articles

An early suitability assessment of two exotic Ophraella species (Coleoptera: Chrysomelidae) for biological control of invasive ragweed in Europe

Classical biological control is an important means of managing the increasing threat of invasive plants. It constitutes the introduction of natural enemies from the native range of the target plant into the invaded area. This method may be the only cost-effective solution to control the rapidly expanding common ragweed, Ambrosia artemisiifolia, in non-crop habitats in Europe. Therefore, candidate biocontrol agents urgently need to be assessed for their suitability for ragweed control in Europe. A previous literature review prioritized the host-specific leaf beetle Ophraella slobodkini as a candidate agent for ragweed control in Europe, whereas it rejected its oligophagous congener O. communa. Meanwhile, O. communa was accidentally introduced and became established south of the European Alps, and we show here that it is expanding its European range. We then present a short version of the traditional pre-release risk-benefit assessment for these two candidate agents to facilitate fast decision-making about further research efforts. We selected two complementary tests that can be conducted relatively rapidly and inform about essential risks and benefits. We conducted a comparative no-choice juvenile performance assay using leaves of ragweed and sunflower, the most important non-target plant, in Petri dishes in climatic conditions similar to that in the current European range of O. communa. This informs on the fundamental host range and potential for increasing abundance on these host plants. The results confirm that O. slobodkini does not survive on, and is hence unlikely to cause severe damage to sunflower, while O. communa can survive but develops more slowly on sunflower than on ragweed. In parallel, our species distribution models predict no suitable area for the establishment of O. slobodkini in Europe, while O. communa is likely to expand its current range to include a maximum of 18% of the European ragweed distribution. Based on this early assessment, the prioritization and further assessment of O. slobodkini seem unwarranted whereas the results urgently advocate further risk-benefit analysis of O. communa. Having revealed that most of the European area colonized by ragweed is unlikely to be suitable for these species of Ophraella we suggest the use of such relatively short and cheap preliminary assessment to prioritise other candidate agents or strains for these areas.

Predicting the risk of non-target damage to a close relative of a target weed using sequential no-choice tests, paired-choice tests and olfactory discrimination experiments

ABSTRACTWe investigated host-plant utilisation by the candidate biocontrol agent Paradibolia coerulea (Coleoptera: Chrysomelidae) on the target plant Spathodea campanulata Beauv. (Bignoniaceae) and a closely related non-target plant, Kigelia africana (Lam.) Benth. (Bignoniaceae). Paired-choice and sequential no-choice experiments were performed and coupled with olfactory discrimination experiments to test the insects’ responses to volatiles from both plant species as well as to cues from conspecific beetles. Although K. africana was utilised by P. coerulea, S. campanulata was preferred for both adult feeding and oviposition. Interestingly, whereas females were attracted to olfactory cues emitted by S. campanulata, males demonstrated no such olfactory discrimination. Females were also attracted to cues deposited by males, and males were deterred by cues from other males, but neither sex responded to female olfactory cues. Very few eggs were recorded on K. africana and none of the larvae that hatched on K. africana survived the first instar. Both S. campanulata and K. africana are suitable for adult feeding, but persistent utilisation of K. africana in the field is unlikely because larval development is only possible on S. campanulata and because the adult females are strongly attracted to volatiles emitted by the target plant. Nevertheless, if P. coerulea is released as a biocontrol agent, spill-over adult feeding could potentially occur on K. africana growing sympatrically with S. campanulata. Because P. coerulea cannot complete its development on K. africana, non-target damage will only occur where the target plant is present, with an intensity dependent on densities of adult beetles locally.

Competition assays revealed Paenibacillus pasadenensis strain R16 as a novel antifungal agent

The development of new sustainable containment strategies of plant diseases is very important to guarantee food security while reducing the environmental impact of agriculture. Research of new biocontrol agents is a long and difficult process that involves several steps that start from the identification of possible candidates which, for example, show antibiotic activities, and ends with in field, large scale trials. In this study, the plant growth promoting potential and antifungal effect exerted by a novel, putative candidate biocontrol agent, strain R16, identified as Paenibacillus pasadenensis by sequence analysis of 16S rRNA and rpoB genes, against three important plant pathogenic fungi (Botrytis cinerea, Fusarium verticillioides, and Phomopsis viticola), were assessed. Biochemical assays to determine plant growth promoting potential gave negative results for siderophore production and phosphate solubilization, and positive results for ACC-deamination and IAA production. Further biochemical assays for endophytic lifestyle and antifungal activity gave positive results for catalase and chitinase activity, respectively. In vitro antagonism assays showed that strain R16 is effective against B. cinerea, reducing mycelial growth both in dual-culture and through volatile substances, characterized to be mostly composed by farnesol, and inhibiting conidia germination. Good antagonistic potential was also observed in vitro towards P. viticola, but not towards F. verticillioides. Moreover, in vivo assays confirmed the strain R16 activity reduced the infection rate on B. cinerea-inoculated berries. The obtained results firstly proved that P. pasadenesis strain R16 is a putative plant growth promoter and effective against phytopathogenic fungi. Further studies will be needed to investigate the possible application of this strain as a biocontrol agent.

Community structures and antagonistic activities of the bacteria associated with surface-sterilized pepper plants grown in different field soils.

Endophytic bacteria may act individually or in consortia in controlling certain plant diseases. In this study, pepper plants (Capsicum annuum L. cv. Nokkwang) were cultivated in glasshouse conditions using field soils collected from two different geographic locations, Deokso (DS) and Gwangyang (GY) in Korea. Community structure and antifungal activity of pepper endophytic bacteria were analyzed using culture-independent (PCR-DGGE) and culture-dependent (plating) methods, respectively. Dissimilarities were observed between DGGE profiles of DS and GY samples at all plant tissues. However, sequencing of the major DGGE bands revealed an enrichment of Firmicutes in the leaves of plants propagated in either soil. Similar results were observed with the culturable assays. Firmicutes dominated the isolates from both leaf samples, DS leaf (100%) and GY leaf (83.3%), although the genus compositions of DS leaf and GY leaf isolates were different. We assessed the antifungal activity of each isolate recovered to better understand the potential role that these endophytic bacteria may play. Of the 27 representative isolates from DS plant samples, 17 isolates (63.0%) had antagonistic activity against at least one of the fungi tested. Seventeen isolates from GY plant samples (58.6%) displayed antagonistic properties. The results show that the endophytic communities differ in the same plant species when propagated in different soils. Exploring the internal tissues of plants growing in diverse soil environments could be a way to find potential candidates for biocontrol agents.

Laboratory biology and impact of a stem-boring weevil Apocnemidophorus pipitzi (Coleoptera: Curculionidae) on Schinus terebinthifolia

ABSTRACTBrazilian peppertree, Schinus terebinthifolia Raddi (Anacardiaceae), was introduced into Florida, USA, from South America as an ornamental plant in the late 19th and early 20th centuries. It eventually escaped cultivation and is a serious threat to the state’s biodiversity. In the 1980s, this invasive weed was targeted for classical biocontrol. Surveys for natural enemies of Brazilian peppertree conducted in the native range resulted in the discovery of several candidate biocontrol agents. A stem-boring weevil identified as Apocnemidophorus pipitzi (Faust) was collected in Paraguay and transported under permit to Florida, USA. A laboratory colony of A. pipitzi was established in April 2007 by caging adults on cut branches of Brazilian peppertree supplemented with leaf bouquets. Adults are defoliators that feed mainly on the upper surface of subterminal leaflets. Females deposit eggs singly inside the stems and larvae feed under the bark where they damage the vascular cambium. There are five instars, pupation occurs inside the stem and a new generation is produced in 3–4 months. Growth of potted plants with and without exposure to weevil herbivory was compared over an 11-month period. Feeding damage by adults and larvae significantly increased leaf abscission and reduced leaf and root biomass accumulation.

Identification of pigmented Serratia marcescens symbiotically associated with Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae).

To characterize red pigment‐producing bacteria (RPPB) regularly released during oviposition by red palm weevil (RPW), RPPB were recovered from eggs deposited in apples supplied as substrate for oviposition. The presence of RPPB was also detected from gut, the reproductive apparatus of dissected adult and virgin insects and from pupal cases collected within infested palms. RPPB were also identified all along the tissue of these palms. Analysis of the 16S rDNA, gyrB, rpoB, recA, and groEL sequences assigned RPPB to the species Serratia marcescens. RPPB exhibited an antimicrobial activity assessed by the agar well diffusion method against a number of gram‐positive and gram‐negative bacteria. In this study, we first report the identification of a red pigment‐producing S. marcescens as extracellular symbiont of RPW. Route of transmission, detection within different organs, and a wide spread along the infested palm tissue, suggested S. marcescens is present as extracellular symbiont in different developmental stages of the RPW. Additionally, the antimicrobial activity exhibited versus Bacillus spp., Paenibacillus spp., and Lysinibacillus spp., reported as insect pathogens and potential candidates for biocontrol agents, could ascribe for S. marcescens a potential protective role.

Actinomycetes from Eucalyptus and their biological activities for controlling Eucalyptus leaf and shoot blight

In Thailand, Eucalyptus plantations rapidly expand across the country. Leaf and shoot blight caused by Cryptosporiopsis eucalypti, Cylindrocladium sp. and Teratosphaeria destructans is a serious disease in Eucalyptus plantations. In this study, a total of 477 actinomycete strains were successfully isolated from roots and rhizosphere soil of Eucalyptus. Four hundred and thirty nine isolates were classified as streptomycetes and 38 isolates were non-streptomycetes. Among these isolates, 272 (57.0%), 118 (24.7%) and 241 (50.5%) isolates were antagonistic to Cryptosporiopsis eucalypti, Cylindrocladium sp. and Teratosphaeria destructans, respectively. All isolates were tested for their abilities to produce siderophores, indole acetic acid (IAA) and solubilise phosphate. Most isolates (464, 97.3%) produced siderophores. The majority of isolates (345, 72.3%) solubilised phosphate. In addition, almost half of these isolates (237, 49.7%) produced indole acetic acid. Strain EUSKR2S82 which showed the strongest inhibitory effect against all tested fungi with plant growth promoting ability was selected to test with Eucalyptus. This strain could colonize plant roots and increase Eucalyptus roots length. In a detached leaves bioassay, the disease severity of EUSKR2S82-inoculated Eucalyptus leaves was only 30% compared to 95% in the control treatment. The 16S rRNA gene sequence analysis revealed that the strain EUSKR2S82 was related to Streptomyces ramulosus NRRL-B 2714T (99.44% similarity). Identification of non-streptomycete isolates using 16S rRNA gene sequences classified them into 9 genera: Actinoallomurus, Actinomadura, Amycolatopsis, Cryptosporangium, Microbispora, Micromonospora, Nocardia, Nonomuraea and Pseudonocardia. It is evident that Eucalyptus tree harbored several genera of actinomycetes. The selected isolate, EUSKR2S82 showed potential as a candidate for biocontrol agent of leaf and shoot blight of Eucalyptus and to promote growth.

Population dynamics of Spalgis epius (Lepidoptera: Lycaenidae), a candidate biocontrol agent of mealybugs and its interaction with mealybug-attendant ants

ABSTRACTSpalgis epius is an economically important hemipterophagous butterfly. Detailed information on the population dynamics, natural enemies, and prey range of S. epius and its association with mealybug-attendant ant species is lacking. Three years of field studies conducted at Bangalore University campus, Bengaluru, India on these aspects indicated that the population density of S. epius was greatest from June to December and least from February to May in the low land region. S. epius survived on eight prey species, which were present in different months, of which Phenacoccus indicus was recorded as a prey of S. epius for the first time. Different prey species occurred on 12 species of host plants. The occurrence of S. epius was negatively correlated with temperature and positively correlated with relative humidity and prey populations. Six mealybug-attendant ant species were associated with the larvae of S. epius. The seven general predators of S. epius adults were recorded. Knowledge on the population dynamics and a prey range of S. epius and its interaction with mealybug-attendant ant species could be helpful to using this predator as a major biocontrol agent of various species of mealybugs. This study contributed to our understanding of the population dynamics of a hemipterophagous butterfly.

Relative performance on test and target plants in laboratory tests predicts the risk of non-target attack in the field for arthropod weed biocontrol agents

We tested the hypothesis that quantifying the relative performance of candidate weed biocontrol agents on test and target plants during laboratory host-range testing can predict the probability of test plants being attacked in the field. For arthropod weed biocontrol agents established in New Zealand between 1929 and 2010, the probability of host use was positively correlated with relative performance for both no-choice starvation tests and no-choice oviposition tests. Moreover, multiplying together the relative performance scores for no-choice starvation and oviposition tests to create a combined risk score for no-choice tests resulted in a clear-cut threshold indicating that for scores between 0.21 and 0.33 the probability of host use (including minor spillover attack) occurring rose from close to zero to a virtual certainty. Choice oviposition test data showed a similar pattern to no-choice data, except there were two cases where host use occurred in the field on plant species that had very low relative performance scores in the choice tests. Both of these cases were associated with asynchrony between seed-feeding biocontrol agent activity and the reproductive phenology of the target plant, indicating that choice tests may be inappropriate for certain guilds of biocontrol agent that attack ephemeral plant structures.We conclude that quantitative laboratory testing data can help predict risk of non-target attack. The ability to refer to threshold relative performance scores when deciding whether an agent is safe to release has the potential to ensure fewer environmentally safe candidate biocontrol agents are erroneously rejected, thereby enhancing efficiency in the selection and approval of new agents.

A novel strategy to reduce overwintering inoculum of Monilinia laxa

Brown rot on cherry and plum, caused by Monilinia laxa, is an important disease, for which overwintered mummified fruit is a significant inoculum source for infection of flowers and fruit in the spring. Experiments were conducted to assess the potential of applying plant protection products in winter and/or early spring to suppress sporulation on mummified fruit. Products tested included Indar 5 EW (a commercial fungicide, a.i. fenbuconazole), Aureobasidium pullulans Y126 [a candidate biocontrol agent (BCA)]), Bacillus sp. B91 (a candidate BCA) and Serenade (a commercially formulated BCA of Bacillus subtilis strain QST 713) in addition to control treated with tap water. Indar and A. pullulans Y126 significantly reduced sporulation when applied once in winter. Overall, a single treatment in early spring (February) was slightly more effective than single treatment in winter (November). Application of all products in both winter and early spring led to significant reduction in sporulation. Indar had the highest efficacy, reducing the number of spores from 9 × 105 to 5 × 103 per mummified plum when applied twice. Of the three BCAs applied on both occasions, A. pullulans Y126 had the highest efficacy, reducing the number of spores from 9 × 105 to 5 × 104 per mummified plum. There were no synergistic but additive effects between the two applications in winter and early spring based on the Bliss independence test. These results suggest that reducing overwintering inoculum in dormant season is effective and may be part of an integrated management strategy for brown rot on stone fruit.

POTENSI BAKTERI ENDOFIT DALAM MENEKAN PENYAKIT LAYU STEWART (PANTOEA STEWARTII SUBSP. STEWARTII) PADA TANAMAN JAGUNG

Potential of endophytic bacteria to control stewart wilt disease (Pantoea stewartii subsp. stewartii) in maize. The purpose of this study was to explore endophytic bacteria from seedling, maize roots and grass roots as well as to test the ability of endophytic bacteria which could potentially suppress stewart wilt disease development in maize. Characterization of endophytic bacteria as biocontrol agents including: do not induce HR on tobacco, synthesize IAA, dissolve phosphate, produce siderophores, and antibiotic to Pantoea stewartii subsp. stewartii (Pnss). The results of research shoed 17 isolates of endophytic bacteria potentially as candidate biocontrol agents. Nine isolates were able to produce IAA, siderofores and phosphatase; two isolates produce IAA and phosphatase; six isolates produce IAA. Six isolates ie: AR1, AJ34, AJ15, AJ19, and AJ14 AN6, can increase maize plant resistance and suppress stewart wilt disease severity with a range of 48.95-55.60%.

Dispersal potential of native and exotic predatory ladybirds as measured by a computer-monitored flight mill

The performance of three species of predatory ladybirds was compared in a flight mill and the effect of diet on their flight parameters was tested. The invasive ladybird Harmonia axyridis Pallas (Coleoptera: Coccinellidae) outperformed Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae) and Adalia bipunctata (L.) (Coleoptera: Coccinellidae) in terms of flight distance, duration and velocity. Harmonia axyridis flew at least two times further, needed three times less breaks and flew two times faster than C. montrouzieri and A. bipunctata fed the same diet. Ladybirds reared on eggs of Ephestia kuehniella (Zeller) (Lepidoptera: Pyralidae) performed better than their counterparts reared on natural prey (aphids for H. axyridis and A. bipunctata, mealybugs for C. montrouzieri). The findings of this study indicate that comparative flight studies can be useful to identify candidate biocontrol agents with pronounced dispersal abilities and thus can yield significant evidence to be used in an environmental risk assessment. However, it also demonstrates that variability related to mass rearing conditions should not be ignored when standardizing a risk assessment procedure for candidate biocontrol agents.

Brevibacillus as a biological tool: a short review

The significance of Brevibacillus has been documented scientifically in the published literature and commercially in heterologous recombinant protein catalogs. Brevibacillus is one of the most widespread genera of Gram-positive bacteria, recorded from the diverse environmental habitats. The high growth rate, better transformation efficiency by electroporation, availability of shuttle vectors, production of negligible amount of extracellular protease, and the constitutive expression of heterologous proteins make some strains of this genus excellent laboratory models. Regarding biotechnological applications, this genus continues to be a source of various enzymes of great biotechnological interest due to their ability to biodegrade low density polyethylene, ability to act as a candidate bio-control agent, and more recently acknowledged as a tool for the overexpression. This article reviews the properties of Brevibacillus spp. as better biological tools with varied applications.

Insect herbivores associated with Ludwigia species, Oligospermum section, in their Argentine distribution.

The South American water primroses, Ludwigia grandiflora (Michx.) Greuter & Burdet, L. grandiflora subsp. hexapetala (Hook. & Arn.) G.L. Nesom & Kartesz , Ludwigia peploides (Kunth) P.H. Raven, and L. p. subsp. montevidensis (Spreng.) P.H. Raven (Onagraceae, Section Oligospermum), have become invasive in several watersheds of the United States and Europe. Surveys were carried out in center-east of Argentina to find insect species that might serve as biological control agents for L. g . subsp. hexapetala in California and elsewhere. Stems (0.5–0.6 m) of Ludwigia species, Sect. Oligospermum, were collected in 41 sites and analyzed in the laboratory; immature insects were reared to adults. The plant species found in the area were L. grandiflora (2 sites) , L. g. subsp. hexapetala (33 sites), and L. p. subsp. montevidensis (4 sites). There was a variety of insect guilds feeding on L. g. subsp. hexapetala , including six species with stem-borer larvae, one species with fruit-feeding larvae, four species with defoliating larvae, two species with defoliating larvae on young leaves and axil meristems, one species of cell content feeder, and three species of sap feeders. Nine of these species also have defoliating adults. Biological information on most of them is provided. Of these insect herbivores, only two species were also found on L. grandiflora , and one on L. peploides . Several of the species found on L. g. hexapetala , such as the cell-content feeder Liothrips ludwigi (Thysanoptera), the stem-borers Merocnemus binotatus (Boheman) and Tyloderma spp. (Coleoptera), are promising candidates for biocontrol agents.

Isolation and characterization of soil Streptomyces species as potential biological control agents against fungal plant pathogens

The use of antagonist microorganisms against fungal plant pathogens is an attractive and ecologically alternative to the use of chemical pesticides. Streptomyces are beneficial soil bacteria and potential candidates for biocontrol agents. This study reports the isolation, characterization and antagonist activity of soil streptomycetes from the Los Petenes Biosphere Reserve, a Natural protected area in Campeche, Mexico. The results showed morphological, physiological and biochemical characterization of six actinomycetes and their inhibitory activity against Curvularia sp., Aspergillus niger, Helminthosporium sp. and Fusarium sp. One isolate, identified as Streptomyces sp. CACIS-1.16CA showed the potential to inhibit additional pathogens as Alternaria sp., Phytophthora capsici, Colletotrichum sp. and Rhizoctonia sp. with percentages ranging from 47 to 90%. This study identified a streptomycete strain with a broad antagonist activity that could be used for biocontrol of plant pathogenic fungi.

Lysobacter capsici YS1215를 이용한 뿌리혹선충(Root-knot nematode)의 생물학적 방제

Lysobacter capsici YS215의 특성 및 뿌리혹선충 방제에 미치는 영향을 조사하였다. YS1215의 생육은 배양 6일째 최고였으며, 생육에 따른 chitinase와 gelatinase의 활성은 각각 3일째와 5일째에 가장 높은 활성을 보였다. YS1215 배양액이 선충 피해 방제와 식물 생장에 미치는 영향을 조사해 본 결과, 5주째 식물 지상부 생체중 및 건조중에서 배양액 반량구에서 가장 높게 나타났지만, 9주째에는 미생물 배양액, 미생물 배양액 반량구 및 배지액 처리구에서 차이를 보이지 않았다. 하지만 9주째 미생물 배양액, 미생물 배양액 반량구 및 배지액 처리구가 물처리구 보다 높게 나타났다. 지상부 길이에서는 미생물 배양액 반량구 처리구가 가장 높았다. 선충 피해 방제에 있어서 난낭수, 뿌리혹수 및 토양내 유충수에서 각각 농약 처리구에서 가장 낮게 나타났으나, 미생물 배양액 처리구와의 유의적 차이는 보이지 않았다. 미생물 배양액 처리구는 미생물 배양액 반량구 및 물 처리구와는 유의적 차이가 있는 것으로 조사되었다. 그러므로 다양한 분해효소를 생성하는 L. capsici YS1215의 뿌리혹선충방제에 대한 충분한 가능성과 가치가 있다고 사료된다. The experiments were carried out to investigate the biocontrol potential of Lysobacter capsici YS1215 on root-knot nematode and to characterize its lytic enzyme activities. L. capsici YS1215 showed chitinase and gelatinase activities on the medium containing 0.5% chitin or 0.5% gelatin as substrates. Cell growth of L. capsici YS1215 was highest at 6 days, and the highest activities of chitinase (4.0 unit <TEX>$ml^{-1}$</TEX>) and gelatinase (7.43 unit <TEX>$ml^{-1}$</TEX>) were observed on 3 and 5 days after incubation, respectively. To investigate the effect of L. capsici YS1215 on tomato growth and nematode infection, the plants in pot trial were treated with bacterial culture (BC), half of bacterial culture (HBC), only bacterial medium (BM), tap water (TW) and commercial nematicide (CN). HBC treatd plants showed the higher shoot fresh weight and dry weight on <TEX>$5^{th}$</TEX>week after incubation while BM, HBC and BC had consistently higher values than TW at <TEX>$9^{th}$</TEX> week. HBC appeared to be the highest shoot fresh length at <TEX>$9^{th}$</TEX> week. Both CN and BC showed lower number of egg mass, root gall, and population of juveniles in soil compared to BC, HBC, BM and TW. These results suggest that L. capsici YS1215 with its strong ability of lytic enzyme production can be one of the most significant candidates for biocontrol agents against root-knot nematodes.

Genome sequence reveals that Pseudomonas fluorescens F113 possesses a large and diverse array of systems for rhizosphere function and host interaction

BackgroundPseudomonas fluorescens F113 is a plant growth-promoting rhizobacterium (PGPR) isolated from the sugar-beet rhizosphere. This bacterium has been extensively studied as a model strain for genetic regulation of secondary metabolite production in P. fluorescens, as a candidate biocontrol agent against phytopathogens, and as a heterologous host for expression of genes with biotechnological application. The F113 genome sequence and annotation has been recently reported.ResultsComparative analysis of 50 genome sequences of strains belonging to the P. fluorescens group has revealed the existence of five distinct subgroups. F113 belongs to subgroup I, which is mostly composed of strains classified as P. brassicacearum. The core genome of these five strains is highly conserved and represents approximately 76% of the protein-coding genes in any given genome. Despite this strong conservation, F113 also contains a large number of unique protein-coding genes that encode traits potentially involved in the rhizocompetence of this strain. These features include protein coding genes required for denitrification, diterpenoids catabolism, motility and chemotaxis, protein secretion and production of antimicrobial compounds and insect toxins.ConclusionsThe genome of P. fluorescens F113 is composed of numerous protein-coding genes, not usually found together in previously sequenced genomes, which are potentially decisive during the colonisation of the rhizosphere and/or interaction with other soil organisms. This includes genes encoding proteins involved in the production of a second flagellar apparatus, the use of abietic acid as a growth substrate, the complete denitrification pathway, the possible production of a macrolide antibiotic and the assembly of multiple protein secretion systems.

Extracellular enzyme production and fungal mycelia degradation of antagonistic Streptomyces induced by fungal mycelia preparation of cucurbit plant pathogens

Streptomyces are beneficial soil microorganisms and potential candidates for biocontrol agents against soilborne pathogenic fungi of cucurbit plants. Extracellular enzymes such as cellulase, chitinase and glucanase produced by Streptomyces are important components of actinomycete-fungus antagonism. This study aimed to investigate the influence on extracellular enzymes production and fungal mycelia degradation by antagonistic Streptomyces of mycelia preparation of pathogenic fungi (MPPF) of cucurbit plants. The results showed that the antagonistic Streptomyces displayed high extracellular enzyme activities to varying degrees when MPPF was used as the sole carbon source. The MPPF from Fusarium proliferatum, Fusarium oxysporum f. sp. niveum and Alternaria tenuissima were the most effective carbon sources in enhancing the cellulase activity of Streptomyces globisporus C7, Streptomyces globisporus subsp. globisporus C28 and Streptomyces kanamyceticus C49, respectively. S. globisporus subsp. globisporus C28, Streptomyces pactum A12 and S. kanamyceticus C49 cultured in the medium containing MPPF from Fusarium equiseti showed the highest chitinase activity (12.35, 12.50 and 15.06 U, respectively) of all the MPPF treatments. Glucanase activity of Streptomyces carnosus A11 was enhanced greatly (9.26 U) when MPPF from A. tenuissima was used as the sole carbon source. A hyphal intertwining and degradation phenomenon was observed when the antagonistic Streptomyces came across the pathogenic fungal mycelia, which was due to a synergistic effect of the extracellular enzymes produced by the antagonistic Streptomyces.

Distribusi Geografis Tungau Parasit Nyamuk Aedes Sp. di Daerah Endemis Demam Berdarah Dengue di Propinsi Jawa Tengah

The geographical distribution patterns of parasitic mites determine the types of parasitic mites that should be developed in these endemic geographical regions. Types of parasitic mites and the patterns of geographic distribution of parasitic mites of Aedes sp. in dengue endemic areas in Central Java Province have not been determined yet. The purpose of this research was to determine the parasitic mite and geographical distribution patterns in dengue endemic areas in Central Java Province. All stages of the experiment were conducted by using a survey method with random sampling technique. Sampling area for Semarang City included the counties of Central Semarang and Tembalang, whereas for Banjarnegara including the villages of Krandegan, Kutabanjar, Parakancanggah, and Sokanandi, and the county of Karanganyar City, Karanganyar. Larvae, pupae and adult stages of Aedes mosquitoes. were sampled during the rainy season. Types of parasitic mites were identified using Walter & Proctor (1999), Pesic (2003) and Gerecke (2004) references. The average value and variance were analyzed for the number of each stage to determine the geographic distribution pattern of parasitic mites. The infection ability was determined by the average intensity of parasitic mites and the prevalence value. The results demonstrated that the families of parasitic mites that infect the larvae of Aedes sp. in dengue endemic areas in Central Java Province were Pionidae 1, Histiostomatidae, Hydryphantidae, Hydrachnidae 1, Arrenuridae, Hydrachnidae 2 and Pionidae 2. Based on the frequency of occurrence, prevalence and the ability to infect, it was concluded that family Histostomatidae is a potential candidate for biocontrol agent of Aedes sp larvae. The distribution pattern of parasitic mite families in endemic areas of dengue fever in Central Java Province, showed a random distribution pattern.

Effect of a Novel Podophage AB7-IBB2 on Acinetobacter baumannii Biofilm

Biofilm formation in Acinetobacter baumannii is a common cause of nosocomial infections in humans. Clinical devices and abiotic surfaces are important sites of colonization leading to formation of biofilms. Such infections are often resistant to multiple antibiotic therapies, and hence there is need for an effective mode of control. Herein, we describe the isolation, characterization of a new lytic bacteriophage of A.baumannii and its effect on biofilm. The phage AB7-IBB2, with a genome size of about 170kb was identified to be of family Podoviridae as revealed by transmission electron microscopy. It had an isometric head (35nm) and a short tail (7nm). It lysed 19/39 (49%) clinical isolates of A.baumannii. Rapid adsorption (>99% adsorbed in 4min), a latency period of 25min and a burst size 22PFU/infected cell was observed. The phage could inhibit A.baumannii biofilm formation and disrupt preformed biofilm as well. The phage has promising potential to be considered as a candidate biocontrol agent for A.baumannii infections.