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As opposed to single-cell yeast and mammalian cell lines, apoptosis has not been greatly investigated in filamentous fungi because antibodies to the relevant fungal apoptosis-related proteins are not available commercially and because multicellular organisms cannot be studied using flow cytometry. Here we demonstrate how antibodies from a nonfungal source could be used to investigate this pathway. We show that apoptosis in the filamentous fungus Botrytis cinerea is triggered by the mitochondria-mediated caspase pathway, with release of the apoptotic factors cytochrome c, caspase 3, and caspase 9, on treatment with Trichoderma harzianum-derived L-amino acid oxidase., (Copyright © 2011 Elsevier Inc. All rights reserved.)

The main purpose for this study is to observe the seasonal and month variations for particulates-bound mercury Hg(p) in total suspended particulates (TSP) concentration, dry deposition at five characteristic sampling sites during years of 2009-2010 in central Taiwan. The results show that the highest and lowest monthly average particulates-bound mercury Hg(p) concentrations in TSP were occurred in Dec. and Oct. at Gao-mei (wetland), Chang-hua (downtown) and He-mei (residential) sampling site. In addition, the results show that the highest and lowest monthly average particulates-bound mercury Hg(p) dry deposition was occurred in Feb. and Oct. at Quan-xing (industrial) sampling site. This study reflected that the mean highest particulates-bound mercury Hg(p) concentrations in TSP and mean highest particulates-bound mercury Hg(p) dry deposition were occurred at Gao-mei (wetland) and Quan-xing (industrial). However, the mean lowest particulates-bound mercury Hg(p) concentrations in TSP and mean lowest particulates-bound mercury Hg(p) dry deposition were also occurred at Gao-mei (wetland). Regarding seasonal variation, the order of mean-particulates-bound mercury Hg(p) concentrations in TSP in winter and spring were Gao-mei (wetland) > Quan-xing (industrial) > Bei-shi (suburban/coastal) > Chang-hua (downtown) > He-mei (residential). Finally, the order of mean-particulates-bound mercury Hg(p) dry deposition in fall, spring and summer were Quan-xing (industrial) > Bei-shi (suburban/coastal) > Chang-hua (downtown) > He-mei (residential) > Gao-mei (wetland).

L-amino acid oxidases (L-AAOs) have been isolated from many organisms, such as snake, and are known to have antibacterial activity. To the best of the authors' knowledge, this is the first report of the cloning of cDNA encoding a novel Trichoderma harzianum ETS 323 L-amino acid oxidase (Th-L-AAO). The protein was overexpressed in Escherichia coli and purified to homogeneity. Comparisons of its deduced amino acid sequence with the sequence of other L-AAOs revealed the similarity to be between 9 and 24%. The molecular mass of the purified protein was 52 kDa, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme substrate specificity was highest for L-phenylalanine, and its optimal pH and temperature for activity were 7 and 40 °C, respectively; exogenous metal ions had no significant effect on activity. Circular dichroism spectroscopy indicated that the secondary structure of Th-L-AAO is composed of 17% α-helices, 28% β-sheets, and 55% random coils. The bacterially expressed Th-L-AAO also mediated antibacterial activity against both gram-positive and gram-negative food spoilage microorganisms. Furthermore, a three-dimensional protein structure was created to provide more information about the structural composition of Th-L-AAO, suggesting that the N-terminal sequence of Th-L-AAO may have contributed to the antibacterial activity of this protein.

Although L-amino oxidase (LAAO; EC 1.4.3.2) has been reported to be a potent antibacterial agent, the mechanism responsible for its antibacterial activity has not been identified. The present study aimed to identify the mechanism responsible for the antibacterial activity of Th-LAAO, an LAAO recently isolated from the extracellular proteins of Trichoderma harzianum ETS 323, at the same time as elucidating the nature of this enzyme. The results obtained indicate that the enzyme activity and structure of Th-LAAO are stable at pH 6-8 and less stable at both pH 4-5.5 and pH 9. At pH 7.0, the optimum temperature for Th-LAAO was found to be 40 °C, comprising the temperature at which enzymatic activity is greatest, with enzymatic activity deceasing with further increases in temperature as a result of thermal denaturation of the enzyme, leading to partial denaturation at 50 °C. The results obtained by confocal microscopy and flow cytometry indicate that Th-LAAO interacts with bacteria to cause membrane permeabilization, and this interaction may be promoted by the amphipathic sequence in Th-LAAO and other cytotoxic LAAOs located at the N-terminus. The findings of increased exogenous H(2) O(2) production and reactive oxidative species accumulation in Th-LAAO-treated bacteria indicate that reactive oxidative species accumulation may trigger forms of cell damage, including lipid peroxidation and DNA strand breakage that results in bacterial growth inhibition. Taken together, the results indicate that the processes of bacterial interaction, membrane permeabilization and H(2)O(2) production are involved in the mechanism responsible for the antibacterial activity of Th-LAAO., (© 2011 The Authors Journal compilation © 2011 FEBS.)

Trichoderma spp. are used as biocontrol agents against phytopathogens such as Rhizoctonia solani, but their biocontrol mechanisms are poorly understood. A novel L-amino oxidase (Th-LAAO) was identified from the extracellular proteins of Trichoderma harzianum ETS 323. Here, we show a FAD-binding glycoprotein with the best substrate specificity constant for L-phenylalanine. Although the amino acid sequence of Th-LAAO revealed limited homology (16-24%) to other LAAO members, a highly conserved FAD-binding motif was identified in the N-terminus. Th-LAAO was shown to be a homodimeric protein, but the monomeric form was predominant when grown in the presence of deactivated Rhizoctonia solani. Furthermore, in vitro assays demonstrated that Th-LAAO had an antagonistic effect against Rhizoctonia solani and a stimulatory one on hyphal density and sporulation in T. harzianum ETS 323. These findings further our understanding of T. harzianum as a biocontrol agent and provide insight into the biological function of l-amino acid oxidase.

The structure of nanocomposite thin films composed of polystyrene-b-poly(2 vinylpyridine)/Au nanoparticles (PS-PVP/Au) during chloroform vapor annealing was investigated. Our results revealed that the morphology of these composites depends on particle size and volume fraction. During solvent annealing, particles segregate to both polymer/substrate and air/polymer interfaces. The segregation of particles to the polymer/substrate interface balances the preferential interaction of two domains to the substrate, resulting in the transition of ordered domains from parallel to normal to the substrate. The introduction of large particles facilitates the segregation of particles to the top of the film. When the volume fraction of segregated particles is high, particles are repelled from the preferred domains to form particle-rich regimes. The wetting behavior of a thin film could be improved by the presence of nanoparticles. Our results showed that small particles provide the better ability to prevent thin film from dewetting., (© 2011 American Chemical Society)

Trichoderma harzianum ETS 323 secretes two glucanases, a 23.5 kDa endoglucanase (EG Th1) and a 61 kDa exoglucanase (ExG Th1). They were identified by their hydrolysis products and were purified to homogeneity. The optimal temperature and pH for both EG Th1 (7.3-fold purification, 5.0% yield) and ExG Th1 (33.7-fold purification, 0.15% yield) were 50 °C and pH 4.5, respectively. The kinetic parameters of EG Th1 (K(m) = 23 mg mL(-1), V(max) = 294 μM min(-1), specific activity = 7.4 U mg(-1)) and ExG Th1 (K(m) = 85 mg mL(-1), V(max) = 385 μM min(-1), specific activity = 24.6 U mg(-1)) toward carboxymethyl cellulose were determined. Both enzymes favored CMC and maintained 100% activity for 10 days at 38 °C. KCl, MgCl(2), HgCl(2), and FeCl(3) showed approximately 30% inhibition against EG Th1 but not ExG Th1. They catalyzed transglycosylation of glucose in the presence of cellobiose, but ExG Th1 exhibited better activity and higher product diversity.

Niosomes are synthetic membrane vesicles formed by self-assembly of nonionic surfactant, often in a mixture with cholesterol and dicetyl phosphate. Because of their inner aqueous core and bilayer membrane shell, niosomes are commonly used as carriers of treatment agents for pharmaceutical and cosmetic applications or contrast agents for clinical imaging applications. In those applications, niosomes are considered as a more economical and stable alternative to their biological counterpart (i.e., liposomes). However, conventional bulk method of niosome preparation requires bulk mixing of two liquid phases, which is time-consuming and not well-controlled. Such mixing conditions often lead to large niosomes with high polydispersity in size and thus affect the consistency of niosome dosage or imaging quality. In this study, we present a new method of niosome self-assembly by microfluidic hydrodynamic focusing to improve on the size and size distributions of niosomes. By taking advantage of the rapid and controlled mixing of two miscible fluids (i.e., alcohol and water) in microchannels, we were able to obtain in seconds nanoscaled niosomes with approximately 40% narrower size distributions compared to the bulk method. We further investigated different parameters that might affect on-chip assembly of niosomes, such as (1) conditions for the microfluidic mixing, (2) chemical structures of the surfactant used (i.e., sorbitan esters Span 20, Span 60, and Span 80), and (3) device materials for the microchannel fabrication. This work suggests that microfluidics may facilitate the development and optimization of biomimetic colloidal systems for nanomedicine applications.

Controlled delivery of therapeutic agents from medical devices can improve their safety and effectiveness in vivo, by ameliorating the surrounding tissue responses and thus maintaining the functional integrity of the devices. Previously, we presented a new method for providing simultaneous controlled delivery from medical devices, by surface assembly of biodegradable polymer nanoparticles (NPs) encapsulating fluorescent dyes. Here, we continue our investigation with NPs loaded with therapeutic agents, dexamethasone (DEX) or plasmid DNA, and evaluated the bioactivity of the released molecules with macrophage cells associated with inflammation. Over a period of one week, NPs encapsulating DEX released 24.9+/-0.8ng from the probe surface and was successful at suppressing macrophage cell growth by 40+/-10%. This percentage of suppression corresponded to approximately 100% drug delivery efficiency, in comparison with the unencapsulated drug. DNA NP coatings, in contrast, released approximately 1ng of plasmid DNA and were effective at transfecting macrophage cells to express the luciferase gene at 300+/-200 relative luminescence/mg total protein. This amount of luciferase activity corresponded to 100% gene delivery efficiency. Thus, NP coatings were capable of providing continuous release of bioactive agents in sufficient quantities to induce relevant biological effects in cell culture studies. These coatings also remained intact, even after 14 days of incubation with phosphate buffered saline. Although the maximum loading for NP coatings is inherently lower than the more established matrix coating, our study suggests that the NP coatings are a more versatile and efficient approach toward drug delivery or gene delivery from a medical device surface and are perhaps best suited for continuous release of highly potent therapeutic agents., (Copyright 2010 Elsevier Ltd. All rights reserved.)

Fungal mycelia mass and pigments are major obstacles to investigating the secretion of bioactive substances such as enzyme activities using a plate assay. In this study, we applied a cellophane membrane and demonstrated that it can block mycelia mass and conidia (especially pigmented spores that would likely interfere with any subsequent color development-based activity detection) while allowing secreting enzymes to pass through. Visual observation after lifting the cellophane membrane and the collected mycelia and conidia indicated that the bioactivities on specific plates were improved significantly, although some fungal growth hurdle was noted. This proved to be true whether the assays were color development based or not., (Copyright 2009 Elsevier Inc. All rights reserved.)

A simple strategy to improve the particle dispersion and structural ordering of magnetic nanoparticle/block copolymer composites is demonstrated. This method consists of manipulating both the block copolymer structure in a solvent and the relative interactions of particle/solvent and polymer/solvent to facilitate the self-assembly of particles in the preferred domain of the block copolymer. In a neutral solvent, the freely expanded structure of the polymer and the slightly weaker affinity between particle and solvent than between particle and polymer promotes the selection of particles in the preferred domain of the block copolymer. Furthermore, more particles are allowed to be incorporated into the polymer matrix while still obtaining the nicely ordered structure when these particles exhibit smaller magnetization.

Cell and tissue responses to implanted biomaterials often limit their effectiveness and lifetime. This is particularly true for materials implanted into the brain. We present here a new approach for the modification of materials to enable release of multiple agents, which might be useful in modulating tissue responses, without changing the properties of the underlying material, in this case, a silicon probe. Poly(lactide-co-glycolide) nanoparticles (NPs) were assembled onto silicon probe surfaces by electrostatic interactions. Charged NPs were fabricated by altering the properties of the surfactant. NPs formed with poly(ethylene-alt-maleic anhydride) (PEMA) were strongly negatively charged; these NPs assembled onto probes best when suspended at nearly physiological conditions (surface density approximately 83,600+/-3000 particles/mm(2)). The percentage of surface area coverage by the NPs was estimated to be approximately 13% and was maintained over two weeks during constant exposure to PBS. Multiple fluorescent NP populations were attached to the same probe to allow visualization of simultaneous delivery of multiple agents by fluorescence microscopy. Release from NP coatings was reproducible and controllable. The distinct release profiles of each agent from the coatings were preserved upon attachment to the surfaces. The unique feature of this new system is that NPs encapsulating various molecules (i.e. drugs, proteins, or DNA) can be fabricated separately, in advance, and simply mixed prior to attachment. The versatility of this delivery system, therefore, makes it suitable for many applications.

As a notable biocontrol agent, Trichoderma harzianum can antagonize a diverse array of phytopathogenic fungi, including Botrytis cinerea, Rhizoctonia solani and Fusarium oxysporum. Elucidating the biocontrol mechanism of T. harzianum in response to the pathogens enables it to be exploited in the control of plant diseases. Two-dimensional gel electrophoresis (2-DE) was performed to obtain secreted protein patterns of T. harzianum ETS 323, grown in media that contained glucose, a mixture of glucose and deactivated B. cinerea mycelia, deactivated B. cinerea mycelia or deactivated T. harzianum mycelia. Selected protein spots were identified using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Ninety one out of 100 excised protein spots were analyzed and some proteins were sequence identified. Of these, one l-amino acid oxidase (LAAO) and two endochitinases were uniquely induced in the media that contained deactivated B. cinerea mycelia as the sole carbon source. Activities of the cell wall-degrading enzymes (CWDEs), including beta-1,3-glucanases, beta-1,6-glucanases, chitinases, proteases and xylanases, were significantly higher in media with deactivated B. cinerea mycelia than in other media. This finding suggests that the cell wall of B. cinerea is indeed the primary target of T. harzianum ETS 323 in the biocontrol mechanism. The possible roles of LAAO and xylanase were also discussed.

The biocontrol fungal species of Trichoderma, which colonizes plant roots, are well-known for their potential to control plant pathogens. Six anthraquinones, of which four have been identified for the first time from Trichoderma and two have already been reported in other strains, were purified from Trichoderma harzianum strain Th-R16 to evaluate their biological activities. The structures of the compounds were determined by one- and two-dimensional NMR. The compounds were shown to exhibit stronger antifungal activity than antibacterial activity. Low yield compounds, like 1,5-dihydroxy-3-hydroxymethyl-9,10-anthraquinone, were found to be more active against fungal pathogens than pachybasin and crysophanol, which were found to be the major extracellular metabolites. Test anthraquinones with higher oxidation numbers had better antifungal activity, and their activities were concentration-dependent.

To elucidate the entire range of proteins that are secreted by Trichoderma harzianum ETS 323 in its antagonism with Rhizoctonia solani, an in vivo interaction between them was mimicked and not only the secreted cell wall-degrading enzymes (CWDEs) but also all of the proteome were investigated. Seven CWDEs, chitinase, cellulase, xylanase, beta-1,3-glucanase, beta-1,6-glucanase, mannanase, and protease,were revealed by activity assay, in-gel activity stain, 2-DE, and LC-MS/MS analysis. Extracellular protein extracts from media that contained R. solani exhibited much higher CWDE activities than media that did not contain R. solani. Cellulase and mannanase activity, however, were insignificant. Activity stain also revealed that beta-1,3-glucanase, beta-1,6-glucanase, and xylanase activity occurred exclusively in media that contained R. solani. Furthermore, 35 of the 43 excised spots on the 2-DE gel were successfully analyzed by LC-MS/MS, and eight proteins were identified. They were two glycoside hydrolases, two proteases, two beta-glucosidases, one endochitinase and, interestingly, one amino acid oxidase. Additionally, a possible mechanism was proposed to elucidate how the cell walls of R. solani are systematically enveloped and disintegrated.

We report on a facile diffusion-based photopatterning technique for generating linear and non-linear decreasing pore-size gradients in cross-linked polyacrylamide gels. Diffusion of low viscosity polymer precursor solutions and a two-step photopatterning process were used to define the decreasing pore-size gradient gels in a microfluidic format, thus eliminating the need for controlled mixing and delivery of polymer precursor solutions. We present an analytical model of the non-steady state diffusion process and numerically evaluate that model for direct comparison with empirical characterizations of the gradient gels. We show that the analytical model provides an effective means to predict the steepness and linearity of a desired gradient gel prior to fabrication. To assess electrophoretic assay performance in the microfluidic gradient gels, on-chip sizing of protein samples (20-116 kDa) was investigated. Baseline resolution of six proteins was demonstrated in 4 s using 3.5% to 10% polyacrylamide gradient gels. The demonstrated ability to conduct efficient protein sizing in ultra-short separation lengths (0.3 cm) means low applied electric potentials are needed to achieve the electric field strengths required for protein separations. The low required electric potentials relax operating constraints on electrical components, as is especially important for translation of the assay into pre-clinical and clinical settings. The gradient gel fabrication method reported is amenable to adaptation to non-sizing protein assays, as well as integration with upstream sample preparation steps and subsequent orthogonal downstream assays.

We investigated the structural features of micelles formed by the self-association of the pentablock copolymer poly[ N,N -(diethyl amino)ethyl methacrylate]-block-poly(ethylene oxide)-block-poly(propylene oxide)-block-poly(ethyleneoxide)-block-poly[ N,N -(diethylamino)ethyl methacrylate] (PDEAEM-PEO-PPO-PEO-PDEAEM) in aqueous solutions by using small-angle neutron scattering SANS. The pentablock copolymer solutions exhibit micellar and gel phases in response to changes in both the temperature and pH by virtue of (1) the lower critical solution temperature of the PPO blocks and (2) the polyelectrolyte character of the pendant PDEAEM blocks. Two modeling schemes were employed to describe the SANS data of semidilute copolymer solutions at higher temperature as they contain interacting charged micelles at pH<7.5 and interacting neutral micelles at higher pH. We have elucidated the structures of the micelles in terms of size, shape, polydispersity, association number, number density, and surface charge. At low pH the charged spherical micelles are less packed with the copolymers presumably due to the electrostatic repulsion between the charged pendant groups. On the other hand, at higher pH the hydrophobic character of the neutral pendant groups enable them to sequester within the micelle core along with the PPO, thus increasing the number density and the core size of the spherical micelles. At higher copolymer concentration reversible thermoresponsive sol-gel transitions were observed at all pH conditions and the rheological behavior of the gels nicely correlates with different organization of micelles with different shapes.

We have investigated the effect of the interfacial interaction on the cross-sectional morphology of the tobacco mosaic virus (TMV) in solution and on two types of solid substrates, SiOx (polar) on Si(100) and polystyrene film (nonpolar) on Si(100), using small-angle X-ray scattering (SAXS) and grazing incidence small-angle X-ray scattering (GISAXS), respectively. Results reveal that the flexible chains at the outer surface of TMV either expand or contract depending on the nature of the substrate. Although the unfavorable interaction between the TMV and the PS causes a minimal effect, the stronger attractive interaction between the outer protein surface of TMV and the SiOx substrate induces pronounced deformation of its cross-sectional morphology.

Anthraquinone-derivatives, chrysophanol and pachybasin, were purified by a silica column chromatography with two different solvent systems from Trichoderma harzianum ETS 323. The fungus was incubated in sugarcane bagasse solid medium at room temperature without rotation. Structure of chrysophanol was solved by X-ray diffraction and pachybasin by NMR spectra. About 233+/-13 mg of pure chrysophanol and 773+/-40 mg of pure pachybasin were recovered per kg of solid cultural medium, with yields 1.7+/-0.2% and 5.6+/-0.5%, respectively.

Convection enhanced drug delivery (CED) is a promising therapeutic method for treating diseases of the brain by enhancing the penetration of drugs. Most controlled release delivery methods rely on diffusion from a source to transport drugs throughout tissue. CED relies on direct infusion of drugs into tissue at a sufficiently high rate so that convective transport of drug is at least as important as diffusive transport. This work describes the fabrication and characterization of microfluidic probes for CED protocols and the role diffusion plays in determining penetration. Microfluidic channels were formed on silicon substrates by employing a sacrificial photoresist layer encased in a parylene structural layer. Flow in the microchannels was characterized by applying constant upstream pressures from 35 to 310 kPa, which resulted in flow rates of 0.5-4.5 microL/min. The devices were used to infuse Evans Blue and albumin in hydrogel brain phantoms. The results of these infusions were compared to a simple convection-diffusion model for infusions into porous media. In vivo infusions of albumin were performed in the gray matter of rats at upstream pressures of 35, 70, and 140 kPa. The microfabricated probes show reduced evidence of backflow along the device-tissue interface when compared with conventional needles used for CED.

As a potential anticarcinogenic agent, polysaccharides from Cordyceps sinensis have been demonstrated to possess strong antioxidation activity. The aim of the present research was to study the optimal medium to produce polysaccharides of C. sinensis by using response surface methodology (RSM). The composition of optimized medium for polysaccharide production calculated from the regression model of RSM was 6.17% sucrose, 0.53% corn steep powder, 0.5% (NH4)2HPO4, and 0.15% KH2PO4 at pH 4.44, with a predicted maximum polysaccharide production of 3.17 g/L. When applying this optimal medium, the maximum polysaccharide production was 3.05 and 3.21 g/L in a shake flask and a 5-L jar fermentor, respectively. When the pH was controlled at a higher level such as pH 5.0, both cell growth and polysaccharide production were inhibited. A low pH of 2.85 was required for maximum production of polysaccharides.

Rice hull ash (RHA) of large surface area was obtained by acid wash and then calcination at 600 degrees C for 4 h. The white ash was then mixed with kaolin and starch to make pellet adsorbents with reasonable strength to be utilized in a packed column. Both ash and pellet samples showed good adsorption capacities toward the organic substances in wastewater. Furthermore, the surface nature of the white ash and pellet adsorbent could be modified through either hydration or esterification reactions. Corresponding changes in silanol concentrations were successfully correlated to changes in adsorption capacity toward either Congo red or vacuum pump oil molecules.

Three starches (maize, rice and wheat), and the two non-starch polysaccharides xanthan and locust bean gum galactomannan (LBG) were examined in gel and dough systems for texture and stability properties during freezing and low temperature storage. Xanthan and LBG were found to confer increased resistance to freeze/thaw cycling on rice starch gels but the non-starch polysaccharides had little effect on the performance of maize and wheat starch gels or on wheat dough.

ABSTRACT A beta-glucuronidase (GUS) reporter gene and a hygromycin B (hygB) phosphotransferase gene were integrated separately into the Trichoderma harzianum strain 1295-22 genome, using biolistic transformation. The mycelial growth and biocontrol ability of the transformed strains did not differ from that of the original strain. The transformed Gus(+)-kanamycin-resistant (Gus(+)Kan(R)) strains were used to monitor growth and interactions with Rhizoctonia solani on creeping bentgrass plants. The hygB-resistant (hygB(R)) strains were used to selectively recover strain 1295-22 from the rhizosphere soil and phylloplane of creeping bentgrass after spray applications. The population levels of two hygB(R) strains and the original strain were very similar for all treatments. All three strains persisted for the duration of the experiment (28 days) in both the rhizosphere soil and on leaves, although population levels declined somewhat over the course of the experiment in unautoclaved soils. In this study, the results demonstrated that hygB(R) strains remained dominant over time when assayed on Trichoderma-selective medium containing hygB. The hygB(R) strains were not displaced by strains that colonized untreated plants. Microscopic observation showed that the Gus(+)Kan(R) strains colonized the rhizoplane, seed coat, and phylloplane of creeping bentgrass. These results supported our earlier observation that strain 1295-22 was rhizosphere and phyllo-plane competent. Interactions between T. harzianum and R. solani were readily observed in situ and changed over time. Two types of reactions were found in these experiments. In the first type, sections of hyphae of R. solani near the hyphae of T. harzianum appeared damaged, and the pathogen appeared necrotic when viewed with a microscope. The second type, observed less frequently than the first type, was typical of myco-parasitism. The findings in this study provide new insight into the interactions between R. solani and T. harzianum, providing a basis for future research.

Trichoderma harzianum strain 1295-22 is an effective biocontrol agent for several fungal diseases. The efficacy of granule and spray applications of strain 1295-22 for control of Pythium root rot, brown patch, and dollar spot of creeping bentgrass was investigated. Spray applications of conidial suspensions (SA) of strain 1295-22 significantly reduced all three diseases of creeping bentgrass turf in both greenhouse and field experiments. Control was greatest when Triton X-100 at 0.1% was added to aqueous spray suspensions. When SA were applied weekly, the biocontrol treatments were equivalent to standard fungicides. Broadcast granule applications (GA) also significantly reduced foliar symptoms of Pythium root rot, dollar spot, and brown patch; turf quality also was enhanced. The populations of Trichoderma spp. in the root zone of a bentgrass putting green treated with SA or GA of strain 1295-22 increased 10- to 100-fold after treatment compared with untreated plots. However, strain 1295-22 was present at high levels on bentgrass leaves only following SA. Collectively, the results suggest that strain 1295-22 possesses both rhizosphere and phylloplane competence. The combination of broadcast applications of granules followed by spray applications of conidia reduced damage from both root and foliar diseases.

Background: A large number of foreign laborers are now working in Taiwan, the majority coming from Southeast Asia where parasitic infections are common. Since 1992, monitoring the intestinal parasitic infections of these foreigners has been carried out, particularly to prevent the introduction of some of the parasites into Taiwan. This paper reports results obtained during the year 1993-1994., Methods: The merthiolate-iodine-formalin (MIF) concentration method was used to detect intestinal parasites; and the data were analyzed by chi 2-test., Results: Among the 3,020 persons examined, 526 (17.4%) were infected and 14 kinds of parasites were identified; i.e. Ascaris lumbricoides, Trichuris trichiura, Strongyloides stercoralis, Enterobius vermicularis, hookworms, Taenia sp., Opisthorchis viverrini, Fasciolopsis buski, Schistosoma japonicum, an echinostome, Entamoeba histolytica. Entamoeba coli, Endolimax nana and Giardia lamblia. By nationality, those workers from Indonesia had the highest infection rate, at 32.3%; they were followed by the Filipinos (18.7%), the Thais (16.6%) and the Malaysians (8.0%). The three or four most prevalent parasites were T. Trichiura (21.8%), A. lumbricoides (5.6%) and hookworms (2.4%) for the Indonesians; T. Trichiura (9.1%), A. lumbricoides (3.7%), hookworms and E. histolytica (2.1% each) for the Filipinos; O. viverrini (8.4%), hookworms (3.3%) and E. histolytica (1.2%) for the Thais; T. trichiura (2.2%), A. lumbricoides and E. histolytica (1.8% each) and hookworms (1.4%) for the Malaysians. Among the 526 persons infected, 92.2% had 1 species, 7.2% had 2 species, 0.4% had 3 species and 0.2% had 4 species of parasites., Conclusions: The present result, a 17.4% infection rate, was significantly higher (p < 0.05) than the 12.3% obtained for the previous year (1992-1993). E. histolytica was found in 34 persons, 16 of whom were working as live-in housemaids. The epidemiological significance of some parasites is discussed here.

Miracidial penetration and intramolluscan development of a zoophilic strain of Schistosoma japonicum were studied in the highly susceptible Oncomelania hupensis chiui and a highly resistant population of Oncomelania hupensis formosana. When exposed to 20 miracidia, an average of 3.6 miracidia penetrated O. h. chiui and 3.4 penetrated O. h. formosana. The distribution of sporocysts in the 2 snail subspecies was similar: 70.4% migrated to the cephalopedal sinus, 14.2% to the anterior viscera, and 3.1-4.9% to the posterior viscera, tentacle, mantle collar, and foot. In O. h. chiui only 1 out of 118 sporocysts degenerated and there was no host cellular response. In O. h. formosana, all the sporocysts survived for 12 hr, but by 5 days postinfection (DPI) all were dead and by 10 DPI they had disintegrated completely; the encapsulation was encountered in only 3 out of 78 sporocysts or 3 out of 23 snails, suggesting that it played a minor role in the defense against S. japonicum. All of the O. h. formosana snails of 10 and 15 DPI showed pronounced leukocytosis and elevated phagocytosis in the heart and increased cell debris in tissue spaces, which coincided with the disappearance of the parasites. It is suggested that these phenomena were provoked in response to degradation products released by the dead parasites.

Echinostoma revolutum from Taiwan was studied in lymneid snails at 29 +/- 0.5 degrees C. Given 3-5 miracidia, 95% of Lymnaea ollula and 40% of Lymnaea swinhoei became positive; the prepatent periods were 18 and 25 days, respectively. The following are based on the observations in Lymnaea ollula: The time required for miracidial penetration was about one hour. The sporocysts developed only in the ventricle of the snail but mother rediae developed in the heart and other organs. Mature daughter rediae were not found in the heart cavity. The sporocysts reached the ventricle within 3 days. Mother rediae were released after 6 days and daughter rediae after 8 days. Given 5 miracidia, 1-3 sporocysts reached the heart and 2-20 mother rediae were found per snail. The number of mature daughter rediae was usually 100-200 although more than a thousand may develop in a snail. The sporocysts and mother rediae attained maximal size 9 days postinfection and started degeneration 13 days postinfection. Daughter rediae were largest at the beginning of cercarial emergence and decreased in size thereafter. Simultaneous production of daughter rediae and cercariae by the mother redia was seen only once in this snail mature cercariae were obtained in 10 days postinfection. The cercariae emerged from a small area of mantle collar near the posterior corner of shell aperture. They were negatively phototactic and positively geotactic. An estimation showed that each snail shed about 350 cercariae a day. The cercariae reached the pericardial cavity of snail in one hour via the renal orifice and metacercariae were seen 4-5 hours after exposure. The infectivity of cercariae at various times after shedding, as expressed by cyst recovery rates, were: 51.6%, O-hr old; 76.1%, 2-hr; 68% 4-hr; 32%, 6-hr; 3%, 8 and 10-hr. Cyst recovery rates were not different within the dosage of 50-500 cercariae per snail. Most metacercariae recovered 1-2 days after cercarial exposure were viable; only 5 among 6,533 cysts were dead.

The literature on schistosomiasis in Ethiopia is reviewed with the objective of bringing together in one paper diverse sources which may not be available to those interested in schistosomiasis. Particular attention is given to the influence of altitude and climate, snail ecology and government economic programs on the distribution of schistosomiasis. Out of 365 communities studied between 1961 and 1986 for Schistosomiasis mansoni, cases were reported from 225 (62%), and in 85 (23%) the prevalence ranged from 10 to 92%. Most transmission sites and S. mansoni infections are in agricultural communities along streams between 1300 and 2000 m altitude infested with Biomphalaria pfeifferi, the major snail intermediate host. S. mansoni transmission above 2200 m and below 800 m is precluded in many parts of Ethiopia by low and high water temperatures, respectively. Schistosomiasis haematobium cases have been reported from 30 of the 54 communities studied, 17 of them with infection rates between 14 and 75%. Endemic S. haematobium appears to be confined in its distribution to lowlands below 800 m altitude. The highly focal distribution of S. haematobium transmission is largely due to the nonsusceptibility of most bulinine snails to the Ethiopian strain of the parasite and low water temperatures in the highlands. Water resources development, resettlement programs, refugee migration and other population movements may result in the spread of endemic S. mansoni. Lack of information on snail host/parasite relationships and the ecology of proven and suspected snail hosts does not permit predictions on the spread of endemic S. haematobium. Past and present schistosomiasis control programs in Ethiopia are reviewed and recommendations made for the national control program.

Schistosoma bovis occurs in at least seven of the 14 rovinces of Ethiopia. Results of faecal and snail surveys in three foci are reported. Adwa. One collection showed that nine out of 26 bulinids were infected with S. bovis. The snail host was a tetraploid form of Bulinus (n = 36). The examination of 200 specimens of cattle faces revealed no S. bovis eggs, which was attributed to poor technique or light infection. Gewani. The snail host was Bulinus abyssinicus, which was also infected with S. haematobium, the combined infection rate being 60%. S. bovis eggs were seen in 1-5% (3/197) of specimens of cattle faeces. Lake Awassa. Among 715 bulinids(a mixture of diploid (n = 18) and tetraploid (n = 36) forms), 22 were infected with S. bovis. Infected snails all belonged to the tetraploid form. Infection in cattle faeces was 5-5%(11/200). The Fasciola infection rates in these three areas were 29%, 78% and 60% respectively. Susceptibility of laboratory and wild animals to the Gewani and Lake Awassa strains of S. bovis was investigated. Combined results show that there are at least five species of wild rodents in Ethiopia which are susceptible to S. bovis: Arvicanthis niloticus, Praomys albipes, Rattus rattus, Mastomys coucha and Lophuromys flavopunctatus, in addition to hamsters, white mice, rabbits and guinea pigs. Immature female worms resembling S. bovis were recovered from a goat and a sheep exposed to a mixture of S. bovis and S. haematobium cercariae shed by naturally infected snails. Using the same mixture of cercariae, a Gelada baboon (Theropithecus gelada) could be infected by both schistosomes, but a dog was completely refractory. ABSCESS. Some of these inconclusive results are thought to be due to a unisexual infection. The Gewani strain of SEWANI STRAIN OF S. bovis had a wider range of snail hosts than the Adwa and Awassa strains, covering the tropicus, truncatus and africanus groups of Bulinus. The Adwa and Awassa strains could infect only members of the truncatus group.

Six new recently identified Taiwan Oncomelania hupensis foci were exposed to miracidia from the Changhua Taiwan zoophilic and Indonesian and Philippine anthropophilic strains of Schistosoma japonicum. All new foci were found susceptible to infection in varying degrees. In general the new foci were more susceptible to the Changhua zoophilic strain of S. japonicum than the anthropophilic strains; 17% exposed to the zoophilic strain shed cercariae while 9% and 7%, respectively, of those exposed to the Indonesian and Philippine human strains demonstrated infection. These snails along with O.h. chiui on Taiwan could possibly serve as intermediate host for human oriental schistosomiasis if the parasite was accidentally introduced onto Taiwan.

Two thousand, three hundred and nine stool specimens from about 5% of the residents of Akaki were examined by Kato thick smear technique for helminthic infections. The prevalence of various parasites was as follows: Schistosoma mansoni, 1.5%; Ascaris lumbricoides, 40.7%; Trichuris trichiura, 27.5%, Enterobius vermicularis, 2.2%; Taenia saginata, 3.2%; and Hymenolepis nana, 0.6%. Infected Biomphalaria pfeifferi snails were collected from the Fanta Stream, and 12% of the residents along the stream were infected with S. mansoni with an arithmetic mean of 437 eggs per gram of faeces (e.p.g.) as compared to 250 e.p.g. average town-wide, indicating that there was active transmission of S. mansoni. The prevalence of S. mansoni infection was low. It is suggested that it would be easier and more cost effective if control measures were applied at this stage.

A survey of intestinal schistosomiasis and vector snails was conducted in the Gumara and Ribb Irrigation project located on the eastern side of Lake Tana, northwest Ethiopia. Within the project area, stool specimens from 1273 people were examined by the thick smear method. The average infection rate for Schistosoma mansoni was 1.6%. Two localities, Addis Zemen and Yifag, located just outside the project area, were also examined, the former showing an S. mansoni prevalence of 34% (75/223) and the latter 18% (18/101) among school children. In Debre Tabor, the provincial capital 50 km away from the project site at an altitude of 2800 m., five S. mansoni cases were found among 111 school children, but they were considered to be imported cases. The possible establishment of intestinal schistosomiasis in the project area is discussed.

2300 persons from 25 communities in Harerge Administrative Region were examined for intestinal helminths. The overall prevalence rates were: Schistosoma mansoni 19.4%, Ascaris lumbricoides 13.4%, Trichuris trichiura 7.9%, and Taenia species 5.8%. Other parasites found in less than 3% included: Hymenolepis nana, Hymenolepis diminuta, Enterobius vermicularis, Strongyloides stercoralis and hookworms. Three age groups, 5-9, 10-14, and 15-19 years, had approximately similar prevalence (34.9-38.8%), but younger children had heavier egg loads (geometric mean egg output per gram of feces (EPG) of 347, 340 and 248, respectively). Boys had a higher prevalence of egg load than girls (42.4%, 395 EPG vs 26.5%, 277 EPG). Infection rates among Moslems were higher than Christians (45.5 vs 23.7%), but they had smaller egg output (333 vs 412 EPG). The frequency of light, moderate and heavy infections showed a normal distribution at 27, 46 and 27%. Biomphalaria pfeifferi was found to be infected with S. mansoni in Hirna (42/250), Esakly (6/80) and Genda Adem (20/200), the latter two in the Erer valley. Urine samples from 8 localities were negative for S. haematobium infection. The need for further study in several river valleys with agricultural potential is suggested.