Comparative biochemical and molecular evaluation of swarming of Proteus and effects of anti-swarm agents

In addition to inadequate understanding of swarming motility and virulence of Proteus, there is paucity of information on the relative effectiveness of the various anti-swarm agents. The anti-swarming effects of urea, sodium dodecylsulphate (SDS) and trihydroxymethylglycine (Tris) on 40 clinical isolates of Proteus Spp. were comparatively investigated and plasmids associated with swarming were characterized. The three substances elicited a comparable concentration-dependent anti-swarming property at 0.25 – 1.25% on nutrient agar. Anti-swarm agents displayed heterogeneity in their ability to cause significant decreases in the expression of virulence factors. Swarm motility was further found to be strongly associated with the expression of virulence factors in these strains. Of the Proteus strains tested, 32 were found to harbour 1 – 4 plasmids of size ranging from 6.0 – 33.5 kb. Plasmid curing resulted in loss of swarming in 65.6% of these strains. In order to reduce the risk of infection with virulent Proteus strains, the laboratory use of urea and SDS is suggested. Key Words: Proteus, swarming, urea, SDS, Tris. African Journal of Biotechnology Vol.3(1) 2004: 99-10

INSECT ANTAGONISTIC BIO-INOCULANTS FOR NATURAL CONTROL OF LEAF-MINING INSECT PESTS OF FRENCH BEANS

Fungal endophytes, specifically, Trichoderma asperellum , Beauveria bassiana and Metarhizium anisopliae ; strains M2RT4, S4SU1 and S4ST7, respectively, are known to deter aphid infestation and damage to leguminous crops, but no research exists on their biochemical profiles and effects on French bean ( Phaseolus vulgaris ) leaf miners. The objective of this study was to assess the ability of these endophytic bio-inoculants to produce extracellular enzymes, as well as determine their impact on French beans seedling emergence and growth, and leaf miner fecundity and pupation. Laboratory and greenhouse experiments were conduct at the International Centre of Insect Physiology and Ecology (ICIPE, Nairobi), and the University of Bonn (Germany). Bio-priming French bean seeds by soaking in fungal suspension containing 108 conidia/ ml for 4 hr positively impacted on seedling vigour. Six weeks after germination, all bio-inoculant treatments improved nodulation by 1.5-1.7 folds, while S4SU1 and M2TR4 improved shoot biomass by ~ 2g. In vitro, spore suspensions of these isolates were highly toxic to leaf miner larvae, often resulting in high mortalities and a suppression of pupation by over 70%. All tested isolates exhibited protease (Index=1.22), lipase (Index=1.32) and cellulase (Index=1.06) activities; with M. anisopliae being the best enzyme secretor when cultured under dark conditions on enzyme-specific substrate for 7-14 days. In vivo B. bassiana-primed seedlings deterred larval feeding and development, resulting in reduced number of larvae, pupae and adults, by 65.3, 43.2 and 54.0%, respectively. For the first time, this study showed that bio-priming French bean seeds with bio-inoculants can boost P. vulgaris growth and nodulation, consequently altering its interaction and response to leaf miner infestation, feeding and damage to treated seedlings. These bio-inoculants, thus, hold great potential as crop protection agents and could be safely used for priming seeds prior to planting, if leaf mining damages and associated yield losses in French beans are to be minimised in an environmentally sound manner. Further studies should be undertaken to elucidate the mechanisms involved in plant growth promotion, nodulation and insect feeding deterrence.Les endophytes fongiques, sp\ue9cialement, Trichoderma asperellum , Beauveria bassiana et Metarhizium anisopliae ; souches M2RT4, S4SU1 et S4ST7, respectivement, sont connues pour dissuader l\u2019infestation et le dommage des aphids sur les plantes l\ue9gumineuses, mais aucune recherche n\u2019a \ue9t\ue9 men\ue9e sur leurs profils biochmiques et leurs effets sur les insectes mineurs de feuilles du haricot vert ( Phaseolus vulgaris ). L\u2019objectif de cette \ue9tude \ue9tait d\u2019\ue9valuer la capacit\ue9 endophytique de ces bio-inoculants \ue0 produire des enzymes extra-cellulaires, ainsi que de d\ue9terminer leurs impacts sur l\u2019\ue9mergence et la croissance des plantules du haricot vert, sur la f\ue9condit\ue9 et la nymphose du mineur des feuilles. Des exp\ue9rimentations de laboratoire et sous serre on \ue9t\ue9 conduites au Centre International de Physiologie et d\u2019Ecologie d\u2019Insecte (ICIPE, Nairobi), et \ue0 l\u2019Universit\ue9 de Bonn (Allemagne). Le bio-amor\ue7age des graines du haricot vert par trempage dans une suspension fongique contenant 108 conidies/ml pendant 4heures a positivement imapct\ue9 la vigueur des plantules. Six semaines apr\ue8s la germination, tous les traitements de bio-inoculants ont augment\ue9 la nodulation par 1,5-1,7 fois, pendant que S4SU1 et M2TR4 ont augment\ue9 la biomasse des pousses d\u2019environ 2g. En conditions in vitro, les suspensions de spores de ces isolats \ue9taient fortement toxiques aux larves des mineurs de feuilles, r\ue9guli\ue8rement provoquant de grandes mortalit\ue9s et une suppression de la nymphose de plus de 70%. Tous les isolats test\ue9s ont produit des activit\ue9s protease (indice=1,22), lipase (indice=1,32) et cellulase (indice=1.06); avec M. anisopliae constiuant le meilleur s\ue9creteur d\u2019enzyme alors qu\u2019il est cultiv\ue9 dans le noir sur un substrat d\u2019enzyme sp\ue9cifique pendant 7-14 jours. En conditions in vitro, les plantules amorc\ue9es avec B. bassiana ont frein\ue9 l\u2019alimentation et le d\ue9veloppement des larves, r\ue9sultant en une r\ue9duction du nombre de larves, nymphes et adultes, par 65,3; 43,2; et 54,0%, respectivement. Pour une premi\ue8re fois, cette \ue9tude a montr\ue9 que le bio-amor\ue7age des graines du haricot vert avec des bio-inoculants peut augmenter la croissance et la nodulation de P. vulgaris, par cons\ue9quent alt\ue8re son interaction et sa r\ue9ponse \ue0 l\u2019infestation, \ue0 l\u2019alimentation de mineur de feuilles et les dommages sur les plantules trait\ue9es. Ces bio-inoculants, constituent ainsi de grands potentiels agents protecteurs de cultures et pourraient \ueatre utilis\ue9s de fa\ue7on s\ue9curisante pour l\u2019amor\ue7age des graines avant leur semis, si les dommages caus\ue9s sur les feuilles et la perte de rendement associ\ue9e sur le haricot vert sont \ue0 r\ue9duire sans danger pour l\u2019environnement. Des \ue9tudes suppl\ue9mentaires devraient \ueatre men\ue9es pour \ue9lucider les m\ue9canismes impliqu\ue9s dans la promotion de la croissance v\ue9g\ue9tale, la nodulation et la dissuasion de l\u2019alimentation de l\u2019insecte

Differentiation of embryonic stem cells through controlled release of growth factors from microspheres

The development of microspheres for the sustained delivery of protein and small drug delivery has been utilised in tissue engineering and drug delivery applications. However problems exist in obtaining a controlled and predictable release pattern of the encapsulated molecules from these materials. In this study, microspheres with a zero order release kinetic profile and no lag phase were developed from a novel PLGA based polymer blend. The novel PLGA based polymer blend was made from blending PLGA with varying compositions of the triblock co-polymer PLGA-PEG-PLGA. These blends were subsequently used in the fabrication of lysozyme and dexamethasone loaded microspheres. Blending of the triblock copolymer with PLGA resulted in a reduction of the glass transition temperature (36.1ºC against 59.7ºC) and an increased mechanical strength (25.25 ± 1.26MPa against 0.26 ± 0.05MPa) for PLGA and 30% triblock w/w microspheres respectively. An incremental increase in the triblock composition within the Triblock/PLGA blends resulted in a corresponding reduction in glass transition temperature of the microspheres. Varying the triblock composition within the microspheres showed no significant effect on entrapment efficiency (EE) of lysozyme (protein) and dexamethasone (drug) within fabricated microspheres (EE ~ 60% for and 75% for loading weight 5% w/w for lysozyme and dexamethasone microspheres respectively). Controlled release experiments showed incorporation of the triblock increased the burst release of the protein and drug molecules from the microspheres and improved their release kinetics, with zero-order release profile (post burst phase) observed at a triblock composition of 30% w/w. A positive correlation between the amount of triblock within the triblock / PLGA blend and the rate of protein and drug release was also observed. The induction of osteogenesis and chondrogenesis within stem cells seeded on dexamethasone and ascorbate phosphate, and TGF-β3 loaded scaffolds was successfully demonstrated. Zonal release of TGF-β3 and BMP4 proteins from a bilayered scaffold was also demonstrated. However experiments conducted to demonstrate the tissue zonation within a bone cartilage bilayered construct developed from embryonic stem cell seeded TGF-β3 and BMP4 loaded bilayered scaffolds yielded inconclusive data. These results suggests that protein and drug loaded injectable microspheres for tissue engineering applications can be formed from triblock/PLGA blends, and that by varying the triblock composition, the temperature at which the microspheres form scaffolds, the release kinetics and the mechanical strength of the resulting scaffolds can be controlled

Langzeit-Systemvergleiche in Kenia und Indien: Konventionelle und biologische Erträge aus dem ersten Umstellungsjahr

Organic agriculture is more and more perceived as a promising approach to increase food security in developing countries. However, only few attempts have been made so far to assess agronomic and economic performance of organic agriculture in these^regions in a systematic way. This article reports the first year’s results of two long-term farming systems comparison field trials in Kenya and India. In sub-humid Central Kenya, on a high potential site in Meru South District (Chuka), there were no differences between yields of conventional and organic systems for the first maize and brassica crops. In contrast, organic yields were 14 to 60% lower than conventional yields on a trial site in a medium potential zone in Maragua District (Thika). It is assumed that the organic crops in Chuka could benefit from N and P mobilisation from the soil. In Thika, where N and P were less available, the crop depended on the easily soluble nutrients applied in the conventional treatments. In the semi-arid cotton belt of^Central India, biodynamic, organic, conventional and genetically modified (GM) cotton are compared. Soya and wheat are also part of the crop rotation under study. Biodynamic and organic cotton and wheat yields were 30% lower than conventional and GM yields. Soya yields did not differ between the treatments. It is suggested that yield in organic farming systems in conversion depends on initial inherent soil fertility and crop

The Challenges of Living with and Caring for a Child or Children Affected by Neuronal Ceroid Lipofuscinosis Type 2 Disease: In-Depth Family Surveys in the United Kingdom and Germany

Limited research has investigated the challenges faced by families caring for children with neuronal ceroid lipofuscinosis type 2 (CLN2) disease. Face-to-face, mixed-method, in-depth surveys were conducted with 19 families (23 children) in the UK (n=9) and Germany (n=10) to assess the impact of caring for children with CLN2 disease, using national wellbeing and quality of life (QoL) measures. Primary (n=19) and secondary (n=10) caregivers, adult siblings (n=2), and child siblings (n=2) were included. Caregivers reported reduced health-related QoL compared with age and gender-matched controls (mean utility scores 0.08 and 0.11 lower in Germany and the UK, respectively). Hours of caregiving were significantly higher relative to that provided to a child of normal health, with stress, back pain, and reductions in sleep being recorded. Lower life satisfaction and happiness with partners were also reported, along with significant financial burden. Those caring for children in the late stage of disease were more greatly impacted than those with children in the rapidly progressive stage, or who were bereaved. The results of this study make clear the importance of emotional and practical support for caregivers and siblings coping with CLN2 disease

Influence of temperature on the development, reproduction and longevity of Ceratothripoides claratris (Thysanoptera: Thripidae) on tomatoes

Ceratothripoides claratris (Shumsher) is a serious pest attacking tomatoes in Thailand. Temperature-dependent development of C. claratris was studied at seven constant temperatures, i.e. 22, 25, 27, 30, 34, 35 and 40°C. Pre-adult survivorship was greatest (95%) at 25 and 30°C and shortest at 22°C. Egg-to-adult time decreased within the range of 20 to 30°C and at 34°C it started to increase. The lower thermal threshold for egg-to-adult development was estimated at 16 and 18°C by linear regression and the modified Logan model, respectively. The optimum temperature for egg-to-adult development was estimated at 32-33°C by the modified Logan model. The influence of temperature on reproduction and longevity of C. claratris was determined at 25, 30 and 35 and 40°C. Both inseminated and virgin females failed to reproduce at 40°C. Virgin females produced only male offspring, confirming arrhenotoky. The sex ratio of the offspring of fertilized females was strongly female-biased, except at 25°C. Mean total fecundity per female and mean daily total fecundity per female were highest for both virgin and inseminated females at 30°C. Female longevity was longest at 25°C and shortest at 40°C. Male longevity was longest at 30°C and shortest at 40°C. The net reproductive rate (R0) and intrinsic rate of natural increase (rm) was greatest at 30°C while, mean generation time (G) and the doubling time (f) were highest at 25°C. The finite rate of increase (λ) was fairly constant (1.1-1.5 days) over the three temperatures tested. The pest potential of C. claratris for tropical Asia is discussed

Investigating health-related quality of life in rare diseases: a case study in utility value determination for patients with CLN2 disease (neuronal ceroid lipofuscinosis type 2)

BACKGROUND: Utility studies enable preference-based quantification of a disease's impact on patients' health-related quality of life (HRQoL). It is often difficult to obtain utility values for rare, neurodegenerative conditions due to cognitive burden of direct elicitation methods, and the limited size of patient/caregiver populations. CLN2 disease (neuronal ceroid lipofuscinosis type 2) is an ultra-rare, progressive condition, for which there are no published utility data fully capturing all disease stages. This case study demonstrates how utility values can be estimated for ultra-rare paediatric diseases by asking clinicians to complete EQ-5D-5L questionnaires based on vignettes describing the stages of CLN2 disease. METHODS: An indirect elicitation method using proxy-reporting by clinical experts was adopted. Eighteen vignettes were developed, describing nine progressive disease stages as defined by motor and language domain scores of the CLN2 Clinical Rating Scale, in individuals treated with cerliponase alfa or standard care. Eight clinical experts with experience of treating CLN2 disease with cerliponase alfa and current standard care completed the proxy version 2 EQ-5D-5L online after reading these vignettes. Resulting scores were converted to EQ-5D-5L utility values for each disease stage, using UK, German and Spanish value sets. RESULTS: Utility values, which are typically anchored by 0 (equivalent to death) and 1 (full health), decreased with CLN2 disease progression (results spanned the maximum range of the utility scale). Assigned utility values were consistently higher for patients receiving cerliponase alfa than standard care; differences were statistically significant for the 6 most severe disease stages (p < 0.05). Analysis of the individual dimensions of the EQ-5D-5L showed that greatest differences between patients treated with cerliponase alfa and standard care occurred in the pain dimension (differences in mean scores ranged between no difference and 1.8), with notable differences also observed in the anxiety/depression dimension (differences in mean scores ranged between 0.1 and 1.0). CONCLUSIONS: This study demonstrates a feasible methodology for eliciting utility values in CLN2 disease, indicating HRQoL declines with disease progression. Vignettes describing patients receiving cerliponase alfa were consistently assigned higher utility values for the same disease state, suggesting this treatment improves HRQoL compared with standard care. Trial registration NCT01907087, NCT02485899

Thermal Tolerance of the Coffee Berry Borer Hypothenemus hampei: Predictions of Climate Change Impact on a Tropical Insect Pest

Coffee is predicted to be severely affected by climate change. We determined the thermal tolerance of the coffee berry borer , Hypothenemus hampei, the most devastating pest of coffee worldwide, and make inferences on the possible effects of climate change using climatic data from Colombia, Kenya, Tanzania, and Ethiopia. For this, the effect of eight temperature regimes (15, 20, 23, 25, 27, 30, 33 and 35°C) on the bionomics of H. hampei was studied. Successful egg to adult development occurred between 20–30°C. Using linear regression and a modified Logan model, the lower and upper thresholds for development were estimated at 14.9 and 32°C, respectively. In Kenya and Colombia, the number of pest generations per year was considerably and positively correlated with the warming tolerance. Analysing 32 years of climatic data from Jimma (Ethiopia) revealed that before 1984 it was too cold for H. hampei to complete even one generation per year, but thereafter, because of rising temperatures in the area, 1–2 generations per year/coffee season could be completed. Calculated data on warming tolerance and thermal safety margins of H. hampei for the three East African locations showed considerably high variability compared to the Colombian site. The model indicates that for every 1°C rise in thermal optimum (Topt.), the maximum intrinsic rate of increase (rmax) will increase by an average of 8.5%. The effects of climate change on the further range of H. hampei distribution and possible adaption strategies are discussed. Abstracts in Spanish and French are provided as supplementary material Abstract S1 and Abstract S2