Commenced in January 2007
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Edition: International
Paper Count: 35

Search results for: obligate parasitism

35 Molecular Identification and Evolutionary Status of Lucilia bufonivora: An Obligate Parasite of Amphibians in Europe

Authors: Gerardo Arias, Richard Wall, Jamie Stevens

Abstract:

Lucilia bufonivora Moniez, is an obligate parasite of toads and frogs widely distributed in Europe. Its sister taxon Lucilia silvarum Meigen behaves mainly as a carrion breeder in Europe, however it has been reported as a facultative parasite of amphibians. These two closely related species are morphologically almost identical, which has led to misidentification, and in fact, it has been suggested that the amphibian myiasis cases by L. silvarum reported in Europe should be attributed to L. bufonivora. Both species remain poorly studied and their taxonomic relationships are still unclear. The identification of the larval specimens involved in amphibian myiasis with molecular tools and phylogenetic analysis of these two closely related species may resolve this problem. In this work seventeen unidentified larval specimens extracted from toad myiasis cases of the UK, the Netherlands and Switzerland were obtained, their COX1 (mtDNA) and EF1-α (Nuclear DNA) gene regions were amplified and then sequenced. The 17 larval samples were identified with both molecular markers as L. bufonivora. Phylogenetic analysis was carried out with 10 other blowfly species, including L. silvarum samples from the UK and USA. Bayesian Inference trees of COX1 and a combined-gene dataset suggested that L. silvarum and L. bufonivora are separate sister species. However, the nuclear gene EF1-α does not appear to resolve their relationships, suggesting that the rates of evolution of the mtDNA are much faster than those of the nuclear DNA. This work provides the molecular evidence for successful identification of L. bufonivora and a molecular analysis of the populations of this obligate parasite from different locations across Europe. The relationships with L. silvarum are discussed.

Keywords: calliphoridae, molecular evolution, myiasis, obligate parasitism

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34 Biotic Potential of Different Densities of Aphid Parasitoids, Diaeretiella rapae (Hymenoptera: Braconidae: Aphidiinae) Feeding on Brevicoryne brassicae

Authors: Muhammad Anjum Aqueel, Muhammad Jaffar Hussain, Abu Bakar Muhammad Raza

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Diaeretiella rapae (M’Intosh) attack most of the aphid species. However, it is specialized in feeding on crucifer aphid, Brevicoryne brassicae. Biological potential of parasitoid is its density-dependency due to sharing of limited resources in few cases. The present study was carried out to check the biotic potential of D. rapae at its different densities (1, 2, 4, 8 and 10 pairs) on fixed number of B. brassicae (100 in number) as a host. The present study was performed under laboratory conditions (25 ± 2 ºC temperature and 65-70 % R.H.). Different biological parameters for parasitoid (e.g. percent parasitism, adult emergence, adult longevity and per pair parasitism) were evaluated to check its biotic potential. The present findings showed that maximum parasitism (43.09 % ± 0.63) was observed in highest density (10 pairs) and minimum parasitism (16.59 % ± 1.28) in lowest density (1 pair) of the parasitoid. Maximum adult emergence (80.31 % ± 1.33) was observed in highest density (10 pairs) and minimum parasitism (45.99 % ± 1.27) in lowest density (1 pair) of the parasitoid. In the case of adult longevity, highest (8.2 days ± 0.38) and lowest (6 days ± 0.32) longevity were observed in lowest (1 pair) and highest (10 pairs) densities of parasitoids respectively. However, per pair parasitism rate decreased with the increase in parasitoid densities due to intra-specific competition, developed between the parasitoids for parasitism. The positive but close relationship was observed between percent parasitism and adult emergence. The increase in parasitoid densities increased the percent parasitism and adult emergence of the parasitoid. So, we conclude that an inter-specific competition negatively affected the efficacy of parasitoids and may reduce the fitness of the emerging parasitoid.

Keywords: Diaeretiella rapae, Parasitoid densities, Percent parasitism, adult emergence

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33 Effect of Different Temperatures and Cold Storage on Pupaes Apanteles gelechiidivoris Marsh (Hymenoptera: Braconidae) Parasitoid of Tuta absoluta Meyrick (Lepidoptera: Gelechiidae)

Authors: Jessica Morales Perdomo, Daniel Rodriguez Caicedo, Fernando Cantor Rincon

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Tuta absoluta known as the tomato leaf miner, is one of the main pests in tomato crops in South America and the main pest in many European countries. Apanteles gelechiidivoris is a parasitoid of third instar Tuta absoluta larvae. Our studies have demonstrated that this parasitoid can cause up to 80% mortality of T. absoluta larvae in the field. We investigated cold storage of A. gelechiidivoris pupae as a method of mass production of this parasitoid. This storage method does not interfere with biological characteristics of the parasitoid. In this study, we evaluated the effect of different temperatures (4, 8 and 12°C) and different time duration (7, 14, 21 or 28 days) of cold storage on biological parameters of A. gelechiidivoris pupae and adults. The biological parameters of the parasitoid evaluated were: adult emergence time, lifespan, parasitism percentage and sex ratio. We found that the adult emergence time was delayed when the parasitoid pupae were stored at 4°C and 8°C. The shortest adult emergence was recorded when pupae were stored for seven days. The lowest adult emergence was found for pupae stored at 4°C and decreased significantly as the days of storage increased. We found high percentages of adult emergence when pupae were stored at 8°C and 12°C for seven days. Adult lifespan decreased with increasing days of cold storage. Adults emerging from pupae stored at 8°C during seven and 14 days showed the longest lifespan (nine days). The lowest parasitism rate was recorded at 4°C at every time point. The highest percentage of parasitism (80%) was found at 8°C during seven days of storage. The treatments had no effect on adults the sex ratio. The results suggest that A. gelechiidivoris pupae can be stored for up to 14 days at 8°C without affecting the efficacy of the parasitoid in the field.

Keywords: biological control, cold storage, massive rearing, quality control

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32 Biophysical Assessment of the Ecological Condition of Wetlands in the Parkland and Grassland Natural Regions of Alberta, Canada

Authors: Marie-Claude Roy, David Locky, Ermias Azeria, Jim Schieck

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It is estimated that up to 70% of the wetlands in the Parkland and Grassland natural regions of Alberta have been lost due to various land-use activities. These losses include ecosystem function and services they once provided. Those wetlands remaining are often embedded in a matrix of human-modified habitats and despite efforts taken to protect them the effects of land-uses on wetland condition and function remain largely unknown. We used biophysical field data and remotely-sensed human footprint data collected at 322 open-water wetlands by the Alberta Biodiversity Monitoring Institute (ABMI) to evaluate the impact of surrounding land use on the physico-chemistry characteristics and plant functional traits of wetlands. Eight physio-chemistry parameters were assessed: wetland water depth, water temperature, pH, salinity, dissolved oxygen, total phosphorus, total nitrogen, and dissolved organic carbon. Three plant functional traits were evaluated: 1) origin (native and non-native), 2) life history (annual, biennial, and perennial), and 3) habitat requirements (obligate-wetland and obligate-upland). Intensity land-use was quantified within a 250-meter buffer around each wetland. Ninety-nine percent of wetlands in the Grassland and Parkland regions of Alberta have land-use activities in their surroundings, with most being agriculture-related. Total phosphorus in wetlands increased with the cover of surrounding agriculture, while salinity, total nitrogen, and dissolved organic carbon were positively associated with the degree of soft-linear (e.g. pipelines, trails) land-uses. The abundance of non-native and annual/biennial plants increased with the amount of agriculture, while urban-industrial land-use lowered abundance of natives, perennials, and obligate wetland plants. Our study suggests that land-use types surrounding wetlands affect the physicochemical and biological conditions of wetlands. This research suggests that reducing human disturbances through reclamation of wetland buffers may enhance the condition and function of wetlands in agricultural landscapes.

Keywords: wetlands, biophysical assessment, land use, grassland and parkland natural regions

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31 Interaction of Dietary Protein and Vitamin E Supplementation on Gastrointestinal Nematode (Gnt) Parasitism of Naturally Infected Lambs

Authors: Ayobami Adeyemo, Michael Chimonyo, Munyaradzi Marufu

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Gastrointestinal nematode (GNT) infection significantly hinder sustainable and profitable sheep production on rangelands. While vitamin E and protein supplementation have individually proven to improve host immunity to parasitism in lambs, to our knowledge, there is no information on the interaction of dietary vitamin E and protein supplementation on lamb growth and GIN faecal egg counts in naturally infected lambs. Therefore, the current study investigated the interaction of dietary protein and vitamin E supplementation on faecal egg counts (FEC) and growth performance of lambs. Twenty four Dohne Merino lambs aged 12 months were allocated equally to each of four treatment combinations, with six lambs in each treatment group for a period of eight weeks. Treatment one lambs received dietary protein and vitamin E (PE), treatment two lambs received dietary protein and no vitamin E (PNE), treatment three received dietary vitamin E and no protein (NPE), and treatment four received no dietary protein and vitamin E supplementation (NPNE). The lambs were allowed to graze on Pennisetum clandestinum contaminated with a heavy load of nematodes. Dietary protein supplementation increased (P < 0.01) average daily gain (ADG) and body condition scores (BCS). Dietary vitamin E supplementation had no effect (P > 0.05) on ADG and BCS. There was no interaction (P > 0.05) between dietary protein and vitamin E supplementation on ADG and BCS. Combined supplementation of dietary protein and vitamin E supplementation significantly reduced (P < 0.01) faecal egg counts and larval counts, respectively. Also, dietary protein and vitamin E supplementation reduced GNT faecal egg counts over the exposure period. The current findings support the hypothesis that the interaction of dietary protein and vitamin E supplementation reduced faecal egg counts and larval counts in lambs. This necessitates future findings on the interaction of dietary protein and vitamin E supplementation on blood associated profiles.

Keywords: gastrointestinal nematodes, nematode eggs, Haemonchus, Trichostrongylus

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30 Endocrine Disruptors Effects on the 20-Hydroxyecdysone Concentration and the Vitellogenin Gene Expression in Gammarus sp.

Authors: Eric Gismondi, Aurelie Bigot-Clivot

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Endocrine disruptors (EDCs) are well known to disrupt the development and the reproduction of exposed organisms. Although this point has been studied in vertebrate models, the limited knowledge of the endocrine system of invertebrates makes the evaluation of EDCs effects difficult. However, invertebrates represent the major part of aquatic ecosystems, such as amphipods Gammaridea, which are crucial for their functioning (e.g., litter degradation, food resource). Moreover, gammarids are hosts of parasites such as vertically-transmitted microsporidia (microsporidia VT), which could be confounding factors in assessment of EDC effects. Indeed, some microsporidia VT could have endocrine effects by their own present in the host since it was observed for example, a feminization of juvenile males, which become phenotypic females. This work evaluated the impact of ethinylestradiol (EE₂, estrogenic), cyproterone acetate (CPA, anti-androgenic), 4-hydroxytamoxifen (4HT, anti-estrogenic) and 17α-methyltestosterone (17MT - androgenic), on the 20-hydroxyecdysone concentration (i.e. 20HE - molt process) and the vitellogenin gene expression (i.e. reproduction) in the freshwater amphipod Gammarus pulex, after a 96h laboratory exposure. In addition, the presence of microsporidia VT was verified in order to analyze the effect of this confounding factor. Results of this study shown that, although endocrine systems of invertebrates and vertebrates are different, EDCs proved in vertebrates could also affect biological functions hormonally controlled in invertebrates. Indeed, the molt process of crustaceans was disrupted in the first stage (i.e. 20-HE concentration) and therefore, could affect, at the long term, the population dynamic. In addition, it was observed that G. pulex was differently impacted according to the gender and parasitism, which underline the importance to take into account these confounding factors to better evaluate the EDCs impact on invertebrate populations.

Keywords: endocrine disruption, gammarus sp., molt, parasitism

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29 Relation Between Demodex Folliculorum and Rosacea

Authors: Maha M. Eissa, Safia M. Ali, Sonia R. Allam, Ezzat A. Badawy, Nahid M. Baddour

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The obligate follicle mite Demodex folliculorum (Df) has been demonstrated in increased numbers in patients with rosacea. The significance of this finding is still not completely understood, because many people free of skin disease may also have mites. So, the aim of this work was to study the possible role of Df in patients with rosacea. Facial punch biopsy specimens were taken from patients with rosacea (either telangiectatic, papulopustular, squamous, granulomatous form or with rhinophyma) and age and sex matched healthy subjects. Results of this study showed that 63.3% of the rosacea patients were harbouring the mite with higher incidence in patients with papulopustular form.

Keywords: Demodex folliculorum, rosacea, papulopustular, punch biopsy

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28 Microbial Contamination of Haemolymph of Honeybee (Apis mellifera intermissa) Parasitized by Varroa Destructor

Authors: Messaouda Belaid, Salima Kebbouche-Gana

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The negative effect of the Varroa bee colony is very important. They cause morphological and physiological changes, causing a decrease in performance of individuals and long-term death of the colony. Indirectly, they weaken the bees become much more sensitive to the different pathogenic organisms naturally present in the colony. This work aims to research secondary infections of microbial origin occurred in the worker bee nurse due to parasitism by Varroa destructor. The feeding behaviour of Varroa may causes damaging host integument. The results show that the microbial contamination enable to be transmitted into honeybee heamocoel are Bacillus sp, Pseudomonas sp, Enterobacter, Aspergillus.

Keywords: honeybee, Apis mellifera intermissa, microbial contamination, Varroa destructor

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27 Impact of ‎Foliar ‎Formulations of Macro and Micro Nutrients on ‎the ‎Tritrophic Association of Wheat Aphid ‎and Entomophagous Insects

Authors: Muhammad Sufyan, Muhammad J. Arif, Muhammad Arshad, Usman Shoukat

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In Pakistan, wheat (Triticum aestivum L.) is seriously attacked by the wheat ‎aphid. Naturally, bio control agents play an important role in managing wheat aphid. However, association ‎among pest, natural enemies and host plant is highly affected by food resource ‎concentration and predator/parasitoid factor of any ecosystem. The present ‎study was conducted to estimate the effect of different dose levels of macro ‎and micronutrients on the aphid population and its entomophagous insect ‎on wheat and their tri-trophic association. The experiment was laid out in ‎RCBD with six different combinations of macro and micronutrients and a control treatment. The data was initiated from the second week of ‎the February till the maturity of the crop. Data regarding aphid population and ‎coccinellids counts were collected on weekly basis and subjected to analysis of ‎variance and mean comparison. The data revealed that aphid ‎population was at peak in the last week of March. Coccinellids population ‎increased side by side with aphid population and declined after second week of ‎April. Aphid parasitism was maximum 25% on recommended dose of Double and ‎Flasher and minimum 8.67% on control treatment. Maximum aphid population was observed on first April with 687.2 specimens. However, this maximum population was shown against the application of Double + Flasher treatment. The minimum aphid population was recorded after the application of HiK Gold + Flasher recommended dose on 15th April. The coccinellids population was at peak level at on 8th April and against the treatment double recommended dose of HiK gold + Flasher. Amount of nitrogen, phosphorus and potassium percentage dry leaves ‎components was maximum (2.33, 0.18 and 2.62 % dry leaves. respectively) in ‎plots treated with recommended double dose mixture of Double + Flasher and ‎Hi-K Gold + Flasher while it was minimum (1.43, 0.12 and 1.77 dry leaves ‎respectively) in plots where no nutrients applied. The result revealed that maximum parasitism was at recommended level of micro and macro nutrients application.‎ Maximum micro nutrients zinc, copper, manganese, iron and boron found with values 46.67 ppm, 21.81 ppm, 62.35 ppm, 152.69 ppm and 36.78 respectively. The result also showed that Over application of macro and micro nutrients should be avoided because it do not help in pest control, conversely it may cause stress on plant. The treatment Double and Flasher recommended dose ratio is almost comparable with recommended dose and present studies confirm its usefulness on wheat.

Keywords: entomophagous insects, macro and micro nutrients, tri-trophic, wheat aphid

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26 Self-Directed-Car on GT Road: Grand Trunk Road

Authors: Rameez Ahmad, Aqib Mehmood, Imran Khan

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Self-directed car (SDC) that can drive itself from one fact to another without support from a driver. Certain trust that self-directed car obligate the probable to transform the transportation manufacturing while essentially removing coincidences, and cleaning up the environment. This study realizes the effects that SDC (also called a self-driving, driver or robotic) vehicle travel demands and ride scheme is likely to have. Without the typical obstacles that allows detection of a audio vision based hardware and software construction (It (SDC) and cost benefits, the vehicle technologies, Gold (Generic Obstacle and Lane Detection) to a knowledge-based system to predict their potential and consider the shape, color, or balance) and an organized environment with colored lane patterns, lane position ban. Discovery the problematic consequence of (SDC) on GT (grand trunk road) road and brand the car further effectual.

Keywords: SDC, gold, GT, knowledge-based system

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25 Inventory Larval Ectoparasites of Tomato Leafminer in National High School of Agriculture, Algeria

Authors: Khadidja Mahdi, Salaheddine Doumandji

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Among the natural enemies that reduce populations of the tomato leaf miner studied in experimental plots of National High school of agriculture (ENSA, Algeria, 36° 40’ à 36° 43’ N.; 3° 08’ à 3° 12’ E.), larval ectoparasites. Three larval ectoparasites are reported in this study namely Necrinmus Sp. and two species of indeterminate Chalcidae (Chalcidae Sp. 1 and 2). These species have significantly reduced the effectives of Tuta absoluta. The results for the parasitism of eggs, larval instars and pupae of Tuta absoluta on the open field tomato in the experimental plots of ENSA show high levels of parasite eggs with 25%. With 94.7%, the first larval instar (L1) is the most parasites. The second instar (L2) undergoes the action of parasitoids least 60%. Instars L3 and L4 and pupae remain unharmed.

Keywords: tuta absoluta, larval ectoparasites, tomato, ensa, Algeria

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24 Time of Release of Larval Parasitoid, Cotesia plutellae (Kurdjumov) on Parasitization of Plutella xylostella L. on Cabbage

Authors: M. T. M. D. R. Perera, N. Senanayake

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Cotesia plutellae is a locally available larval parasitoid of diamondback moth, Plutella xylostella, which can be used to manage P. xylostella in the field in an integrated pest management strategy. A study was undertaken to find out the best time of releasing C. plutellae for effective management of P. xylostella using three release times; 2, 3 and 4 weeks after transplanting of cabbage in farmer’s fields at Marassana in Kandy District, Sri Lanka, during Yala 2014 and 2015 seasons. Results revealed that the percentage mean values of parasitization in Yala 2015, was significantly high; 69.47 and 43.85, when introduced at 2 and 3 weeks after transplanting respectively and significantly low 23.31, when released at 4 weeks after transplanting. It is therefore evident that the parasitoid release should be done before 3 weeks, preferably at 2 weeks after transplanting of cabbage in the field. The highest percentage parasitism achieved was 83.90 at 2 weeks after transplanting in Yala 2015 and the lowest being 18.85 and 12.00% at 4 weeks after transplanting in Yala 2014 and 2015 respectively. Unparasitized larvae were able to maintain high P. xylostella populations up to harvest. Even though there is no yield advantage by using parasitoids for P. xylostella management, the cost incurred for insect pest management was greatly reduced compared to use of synthetic chemicals.

Keywords: cabbage, Cotesia plutellae, larval parasitoid, Plutella xylostella, time of release

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23 Effects on Spiritual Intelligence on Young Adult Muslim Female: Integration of Planned Behaviour Theory in Predicting Consumer Attitude towards Halal Cosmetic

Authors: Azreen Jihan Che Mohd Hashim, Rosidah Musa

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Although 'Spiritual Intelligence' (SI) is hard to measure, it is impossible without a noble value that may affect the attitude in purchasing behavior process, so this paper aims to report on a pilot study analysis results in order to evaluate the degree of SI towards consumers’ attitude in purchasing halal cosmetics and, in turn, to reaffirm intention to purchase by using Theory Planned Behaviour (TPB). It is a descriptive cross-sectional study among the Muslim women as the subjects, working and staying in Klang valley area in Malaysia. The purpose of the study is to develop a new measurement scale to unravel and decompose the underlying dimensions of SI from the perspective of the Muslim deemed imperative. About 200 respondents of users and non-users of halal cosmetics are selected. The structure equation modeling (SEM) was conducted to examine the relationships among god, society and self, which are the dimensions of SI. A finding indicates that, in influencing attitude, those who obligate high spiritual intelligence have a good relationship with god, society and self which may influence them to purchase halal cosmetic product. This study offers important findings and implications for future research as it presents a framework on the importance of SI.

Keywords: spiritual intelligence, god, society, self, young adult Muslim female

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22 Visualization of Interaction between Pochonia Chlamydosporia and Meloidogyne Incognita and Their Impact on Tomato Crop

Authors: Saifullah K., Muhammad Naziruddin Saifullah, Muhammad N.

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The bio control potential and mechanism of P. chlamydosporia against Meloidogyne incognita was evaluated in the present study. Under invitro conditions, P. chlamydosporia was tested for parasitism of eggs and females of M. incognita. The results indicated that this fungus parasitized 87% eggs and 82% females. Culture filtrate (CF) of P. chlamydosporia was tested for its larvicide activity against M. incognita 2nd stage juvenile. The maximum mortality was 97.3% at 100% concentration of the culture filtrate while minimum mortality was 7.3% in 25% concentration after 24 hrs. The result of the pot experiment proved that P. chlamydosporia has reduced the incidence of RKN and improved all tested agronomic growth parameters. The treatment with inoculated M. incognita alone reduced plant height, fresh shoot, and fresh root weight by 44.7%, 29.8%, and 32.8% respectively over uninoculated healthy control. Histopathological studies on the interaction of Pochonia chlamydosporia and Meloidogyne incognita on tomato roots revealed anatomical changes among treatments. Less number of galls with small in size and scarcer abnormalities in the vascular cylinder was observed in plants inoculated with P. chlamydosporia and M. incognita than the plants treated with nematode only. The fungus was seen in in the intercellular spaces of cortical and epidermal cells while the vascular bundles of the plant remain intact, inoculated only with P. chlamydosporia. In the infected roots, many mature females were seen which feed on giant cells. The findings also revealed that control healthy plants were not affected and no histological changes were noted.

Keywords: histopathology, Pochonia chlamydosporia, Meloidogyne incognita, tomato

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21 Designing Garments Ergonomically to Improve Life Quality of Elderly People

Authors: Nagda Ibrahim Mady, Shimaa Mohamed Atiha

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In light of actual needs of elderly people and the changes that accompany age in eyesight, hearing, dexterity, mobility, and memory which make aged people unable to carry out the simplest living affairs especially clothing demands. These needs are almost neglected in the current clothing market obligate aged peoples to wear the available choices without any consideration to their actual desires and needs. Fashion designer has gained many experiences that can gather between ergonomics and stages of fashion designing process. Fashion designer can determine the actual needs of aged people and reply these needs with designs that can achieve Improvement to the life quality of aged people besides maintaining good appearance. Thus Fashion designer can help elderly people to avoid negative impacts age leaves on them, either it is psychological or kinetic or that of dementia. Ergonomics in clothing is considered the tools and mechanisms that are used to fit aged people satisfactions supporting them to improve their living using the least time and effort. Providing the elderly with comfort besides maintaining good appearance that can make self–confidence besides independence. From this point of view the research is looking forward to improve the life of aged people through addressing functional clothes that can make elderly independent in the wearing process. Providing in these designs comfort, quality, and practicality and economic cost. Suggesting the suitable fabrics and materials and applying it to the designs to help the elderly perform their daily living customs. Reaching the successful designs that can be acceptable to specialists and to consumers whom they confirm: it supplies their clothing needs and provides the atheistic and functional performance and therefore it gives them better life.

Keywords: ergonomic, design garments, elderly people, life quality

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20 Biocontrol Effectiveness of Indigenous Trichoderma Species against Meloidogyne javanica and Fusarium oxysporum f. sp. radicis lycopersici on Tomato

Authors: Hajji Lobna, Chattaoui Mayssa, Regaieg Hajer, M'Hamdi-Boughalleb Naima, Rhouma Ali, Horrigue-Raouani Najet

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In this study, three local isolates of Trichoderma (Tr1: T. viride, Tr2: T. harzianum and Tr3: T. asperellum) were isolated and evaluated for their biocontrol effectiveness under in vitro conditions and in greenhouse. In vitro bioassay revealed a biopotential control against Fusarium oxysporum f. sp. radicis lycopersici and Meloidogyne javanica (RKN) separately. All species of Trichoderma exhibited biocontrol performance and (Tr1) Trichoderma viride was the most efficient. In fact, growth rate inhibition of Fusarium oxysporum f. sp. radicis lycopersici (FORL) was reached 75.5% with Tr1. Parasitism rate of root-knot nematode was 60% for juveniles and 75% for eggs with the same one. Pots experiment results showed that Tr1 and Tr2, compared to chemical treatment, enhanced the plant growth and exhibited better antagonism against root-knot nematode and root-rot fungi separated or combined. All Trichoderma isolates revealed a bioprotection potential against Fusarium oxysporum f. sp. radicis lycopersici. When pathogen fungi inoculated alone, Fusarium wilt index and browning vascular rate were reduced significantly with Tr1 (0.91, 2.38%) and Tr2 (1.5, 5.5%), respectively. In the case of combined infection with Fusarium and nematode, the same isolate of Trichoderma Tr1 and Tr2 decreased Fusarium wilt index at 1.1 and 0.83 and reduced the browning vascular rate at 6.5% and 6%, respectively. Similarly, the isolate Tr1 and Tr2 caused maximum inhibition of nematode multiplication. Multiplication rate was declined at 4% with both isolates either tomato infected by nematode separately or concomitantly with Fusarium. The chemical treatment was moderate in activity against Meloidogyne javanica and Fusarium oxysporum f. sp. radicis lycopersici alone and combined.

Keywords: trichoderma spp., meloidogyne javanica, Fusarium oxysporum f.sp.radicis lycopersici, biocontrol

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19 Isolation and Characterisation of Novel Environmental Bacteriophages Which Target the Escherichia coli Lamb Outer Membrane Protein

Authors: Ziyue Zeng

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Bacteriophages are viruses which infect bacteria specifically. Over the past decades, phage λ has been extensively studied, especially its interaction with the Escherichia coli LamB (EcLamB) protein receptor. Nonetheless, despite the enormous numbers and near-ubiquity of environmental phages, aside from phage λ, there is a paucity of information on other phages which target EcLamB as a receptor. In this study, to answer the question of whether there are other EcLamB-targeting phages in the natural environment, a simple and convenient method was developed and used for isolating environmental phages which target a particular surface structure of a particular bacterium; in this case, the EcLamB outer membrane protein. From the enrichments with the engineered bacterial hosts, a collection of EcLamB-targeting phages (ΦZZ phages) were easily isolated. Intriguingly, unlike phage λ, an obligate EcLamB-dependent phage in the Siphoviridae family, the newly isolated ΦZZ phages alternatively recognised EcLamB or E. coli OmpC (EcOmpC) as a receptor when infecting E. coli. Furthermore, ΦZZ phages were suggested to represent new species in the Tequatrovirus genus in the Myoviridae family, based on phage morphology and genomic sequences. Most phages are thought to have a narrow host range due to their exquisite specificity in receptor recognition. With the ability to optionally recognise two receptors, ΦZZ phages were considered relatively promiscuous. Via the heterologous expression of EcLamB on the bacterial cell surface, the host range of ΦZZ phages was further extended to three different enterobacterial genera. Besides, an interesting selection of evolved phage mutants with a broader host range was isolated, and the key mutations involved in their evolution to adapt to new hosts were investigated by genomic analysis. Finally, and importantly, two ΦZZ phages were found to be putative generalised transducers, which could be exploited as tools for DNA manipulations.

Keywords: environmental microbiology, phage, microbe-host interactions, microbial ecology

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18 Nitrogen Fixation in Hare Gastrointestinal Tract

Authors: Tatiana A. Kuznetsova, Maxim V. Vechersky, Natalia V. Kostina, Marat M. Umarov, Elena I. Naumova

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One of the main problems of nutrition of phytophagous animals is the insufficiency of protein in their forage. Usually, symbiotic microorganisms highly contribute both to carbohydrates and nitrogen compounds of the food. But it is not easy to utilize microbial biomass in the large intestine and caecum for the animals with hindgut fermentation. So that, some animals, as well hares, developed special mechanism of contribution of such biomass - obligate autocoprophagy, or reingestion. Hares have two types of feces - the hard and the soft. Hard feces are excreted at night, while hares are vigilance ("foraging period"), and the soft ones (caecotrophs) are produced and reingested in the day-time during hares "resting-period". We examine the role of microbial digestion in providing nitrogen nutrition of hare (Lepus europaeus). We determine the ability of nitrogen fixation in fornix and stomach body, small intestine, caecum and colon of hares' gastro-intestinal tract in two main period of hares activity - "resting-period" (day time) and "foraging period" (late-evening and very-early-morning). We use gas chromatography to measure levels of nitrogen fixing activity (acetylene reduction). Nitrogen fixing activity was detected in the contents of all analyzed parts of the gastrointestinal tract. Maximum values were recorded in the large intestine. Also daily dynamics of the process was detected. Thus, during hare “resting-period” (caecotrophs formation) N2-fixing activity was significantly higher than during “foraging period”, reaching 0,3 nmol C2H4/g*h. N2-fixing activity in the gastrointestinal tract can allocate to significant contribution of nitrogen fixers to microbial digestion in hare and confirms the importance of coprophagy as a nitrogen source in lagomorphs.

Keywords: coprophagy, gastrointestinal tract, lagomorphs, nitrogen fixation

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17 Prey Selection of the Corallivorous Gastropod Drupella cornus in Jeddah Coast, Saudi Arabia

Authors: Gaafar Omer BaOmer, Abdulmohsin A. Al-Sofyani, Hassan A. Ramadan

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Drupella is found on coral reefs throughout the tropical and subtropical shallow waters of the Indo-Pacific region. Drupella is muricid gastropod, obligate corallivorous and their population outbreak can cause significant coral mortality. Belt transect surveys were conducted at two sites (Bohairat and Baydah) in Jeddah coast, Saudi Arabia to assess prey preferences for D. cornus with respect to prey availability through resource selection ratios. Results revealed that there are different levels of prey preferences at the different age stages and at the different sites. Acropora species with a caespitose, corymbose and digitate growth forms were preferred prey for recruits and juveniles of Drupella cornus, whereas Acropora variolosa was avoided by D. cornus because of its arborescent colony growth form. Pocillopora, Stylophora, and Millipora were occupied by Drupella cornus less than expected, whereas massive corals genus Porites were avoided. High densities of D. cornus were observed on two fragments of Pocillopora damicornis which may because of the absence of coral guard crabs genus Trapezia. Mean densities of D. cornus per colony for each species showed significant differentiation between the two study sites. Low availability of Acropora colonies in Bayadah patch reef caused high mean density of D. cornus per colony to compare to that in Bohairat, whereas higher mean density of D. cornus per colony of Pocillopora in Bohairat than that in Bayadah may because of most of occupied Pocillopora colonies by D. cornus were physical broken by anchoring compare to those colonies in Bayadah. The results indicated that prey preferences seem to depend on both coral genus and colony shape, while mean densities of D. cornus depend on availability and status of coral colonies.

Keywords: prey availability, resource selection, Drupella cornus, Jeddah, Saudi Arabia

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16 Seroprevalence of Toxoplasmosis among Hemato-Oncology Patients in Tertiary Hospital of East Cost Malaysia

Authors: Aisha Khodijah Kholib Jati, Suharni Mohamad, Azlan Husin, Wan Suriana Wan Ab Rahman

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Introduction: Toxoplasmosis is caused by an obligate intracellular parasite, Toxoplasma gondii (T. gondii). It is commonly asymptomatic in normal individual, but it can be fatal to immunocompromised patients as it can lead to severe complications such as encephalitis, chorioetinitis and myocarditis. Objective: The aim of the study was to determine the seroprevalence of toxoplasmosis and its association with socio-demographic and behavioral characteristics among hemato-oncology patients in Hospital USM. Methods: In this cross-sectional study, 56 hemato-oncology patients were screened for immunoglobulin M (IgM) antibodies, immunoglobulin G (IgG) antibodies, and IgG avidity of T. gondii by using ELISA Kit (BioRad, USA). For anti-T. gondii IgG antibody, titer ≥ 9 IU/ml was considered as recent infection, while for IgM, ratio ≥ 1.00 was considered as reactive for the anti-T. gondii IgM antibodies. Low avidity index is considered as recent infection within 20 weeks while high avidity considered as past infection. T. gondii exposure, socio-demographic and behavioral characteristics was assessed by a questionnaire and interview. Results: A total of 28 (50.0%) hemato-oncology patients were seropositive for T. gondii antibodies. Out of that total, 27 (48.21%) patients were IgG+/IgM- and one patient (1.79%) was IgG+/IgM+ with high avidity index. Univariate analysis showed that age, gender, ethnicity, marital status, educational level, employment status, stem cell transplant, blood transfusion, close contact with cats, water supply, and consumption of undercooked meat were not significantly associated with Toxoplasma seropositivity rate. Discussion: The seropositivity rate of IgG anti-T. gondii was high among hemato-oncology patients in Hospital USM. With impaired immune system, these patients might have a severe consequence if the infection reactivated. Therefore, screening for anti-T. gondii may be considered in the future. Moreover, health programme towards healthy food and good hygiene practice need to be implemented.

Keywords: immunocompromised, seroprevalence, socio-demographic, toxoplasmosis

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15 Studies on Population and Management of Melon Fruit Fly Bactrocera cucurbitae (Coquillett) in Vegetables Agro-Ecosystem in District Hyderabada

Authors: Abro Zain-Ul-Aabdin, Naheed Baloch, Khuhro Niaz Hussain, Waseem Akbar, Noor Abid Saeed

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The Melon Fruit Fly Bactrocera cucurbitae (Coq.) belongs to family: Tephritidae order: Diptera and is distributed throughout the vegetable growing areas of Pakistan. The B. cucurbitae is injurious pest of more than 125 species of the vegetables throughout the world. In the present studies we investigated the population of this important pest in cucurbit crops and influence of abiotic parameters such as: temperature, relative humidity and rainfall. The study was carried out at two different locations of District, Hyderabad. The locations were Jeay Shah and Dehli farm where three cucurbit vegetable crops, such as bottle gourd (Lagenaria siceraria), bitter gourd (Momordica charantia) and ridge gourd (Luffa acutangula) were grown. The traps were baited with Cue-lure and deployed at three meter height in the all locations from 01.01.2015 and up to 30.06.2015. Results revealed that overall significantly higher (P < 0.05) population was recorded on L.acutangula, M.charantia and L.siceraria (130.64, 127.21, and 122.91), respectively. However, significantly higher (P < 0.05) population was observed on L. acutangula (339.4±22.59) during the 4th week of May 2015 followed by M. charantia (334.6±22.76) L. siceraria (333.2±20.13). Whereas; lowest population was recorded on L. siceraria (5.8±1.39) followed by L. acutangula and M. charantia (6.8±0.80g, 8.0±1.30) respectively during the 4th week of January. The population of B. cucurbitae was significantly correlated with the temperature while negatively correlated with relative humidity. Meanwhile in the parasitism preference experiment pupal parasitoid Dirhinus giffardii showed significantly higher (P<0.05) parasitization when the pupae of B.cucurbitae were reared on Cucumber (Cucumis sativus) (24.8±0.48) and also female were yielded from pupae reared on C.sativus under no choice experiment. Similarly higher parasitization and female were recovered when pupae were supplied C. sativus under free choice experiment. Results of the present investigation would be useful in developing a sustainable pest management strategy in the vegetable agro-ecosystem.

Keywords: Dirhinus giffardii, Bactrocera cucurbitae Cucumis sativus, diptera, free choice, parasitization

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14 Effect of Different Methods to Control the Parasitic Weed Phelipanche ramosa (L. Pomel) in Tomato Crop

Authors: Disciglio G., Lops F., Carlucci A., Gatta G., Tarantino A., Frabboni L, Tarantino E.

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The Phelipanche ramosa is considered the most damaging obligate flowering parasitic weed on a wide species of cultivated plants. The semiarid regions of the world are considered the main center of this parasitic weed, where heavy infestation are due to the ability to produce high numbers of seeds (up to 200,000), that remain viable for extended period (more than 19 years). In this paper 13 treatments of parasitic weed control, as physical, chemical, biological and agronomic methods, including the use of the resistant plants, have been carried out. In 2014 a trial was performed on processing tomato (cv Docet), grown in pots filled with soil taken from a plot heavily infested by Phelipanche ramosa, at the Department of Agriculture, Food and Environment, University of Foggia (southern Italy). Tomato seedlings were transplanted on August 8, 2014 on a clay soil (USDA) 100 kg ha-1 of N; 60 kg ha-1 of P2O5 and 20 kg ha-1 of S. Afterwards, top dressing was performed with 70 kg ha-1 of N. The randomized block design with 3 replicates was adopted. During the growing cycle of the tomato, at 70-75-81 and 88 days after transplantation the number of parasitic shoots emerged in each pot was detected. Also values of leaf chlorophyll Meter SPAD of tomato plants were measured. All data were subjected to analysis of variance (ANOVA) using the JMP software (SAS Institute Inc., Cary, NC, USA), and for comparison of means was used Tukey's test. The results show lower values of the color index SPAD in tomato plants parasitized compared to those healthy. In addition, each treatment studied did not provide complete control against Phelipanche ramosa. However the virulence of the attacks was mitigated by some treatments: radicon product, compost activated with Fusarium, mineral fertilizer nitrogen, sulfur, enzone and resistant tomato genotype. It is assumed that these effects can be improved by combining some of these treatments each other, especially for a gradual and continuing reduction of the “seed bank” of the parasite in the soil.

Keywords: control methods, Phelipanche ramose, tomato crop

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13 Peculiarities of Microflora of Odontogenic Inflammatory Processes in the Central Kazakhstan Region

Authors: Aliya Tokbergenova, Maida Tusupbekova, Daulet Dzhangaliyev, Alena Lavrinenko

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Background: Odontogenic phlegmons are ranked the first among pyoinflammatory processes in the frequency of hospitalization in maxillofacial surgery in the post-Soviet countries. The main role in etiology is played by obligate anaerobes and aerobes. According to numerous data, the structure of aerobic pathogens is dominated by staphylococci and gram-negative bacteria. Aim: The research aim is to study the microflora of the purulent discharge odontogenic inflammatory processes. Materials and methods: A total of 220 patients have been examined, of which 120 patients aged 25-59 years have been included in the research who did not have comorbidity hospitalized in the maxillofacial hospital in Karaganda (Kazakhstan) from January 2016 to July 2017. The bacteriological research has been carried out on the basis of the multiaccess laboratory of the KSMU, through the Matrix Assisted Laser Desorption/Ionization (MALDI) apparatus. The material sample was pus from the inflammation focus, taken during the operating period. Results: According to the research among 120 patients (100%), 15 patients (12.5%) have had microorganisms not grown. From 105 (87.5%) bacteriological results, it has been revealed the following 1) Streptococcus: 51 (42.5%): Streptococcus beta-haemolytic: 17 (14.2%), Streptococcus pneumoniae: 12 (10%), Streptococcus anginosus: 8 (6.6%), Streptococcus oralis: 8 (6.6%), Streptococcus constellatus: 6 (5.0%); 2) Staphylococci: 27 (22.5%): Staphylococci aureus: 14 (11.7%) and Staphylococci epidermidis: 13 (10.8%); 3) Pseudomonas aeruginosa: 12 (10%); 4) Neisseria: 11 (9.1%): Neisseria mucosa: 5 (4.1%) and Neisseria macacae: 6 (5.0%); 5) Klebsiella pneumoniae: 2 (1.7%); 6) Stenotrophomonas maltophilia: 2 (1.7%). 15 patients (12.5%) experienced complications in the form of 1) The dissemination of the process in 10 patients (8.4%). 2) Osteomyelitis in 3 (2.5%). 3) Mediastinitis in 1 (0.8%). 4) Sinusitis in 1 (0.8%). 15 patients (100%) were carried out repeated bacteriological examination, the following was revealed: 1) Streptococcus: 10 (66.7%): Streptococcus beta-haemolytic: 4 (26.7%), Streptococcus pneumoniae: 2 (13.3%), Streptococcus аnginosus: 2 (13.3%), Streptococcus oralis: 1 (6.7%), Streptococcus constellatus: 1 (6.7%); 2) Staphylococci: 4 (26.7%): Staphylococci aureus: 3 (20%) and Staphylococci epidermidis: 1 (6.7%); 3) Pseudomonas aeruginosa: 1 (6.7%). Conclusions: Thus, according to our research data, streptococci predominate in the odontogenic processes microflora in aerobic flora in the central Kazakhstan region, which refutes the leading role of staphylococci in the development of odontogenic inflammatory processes, thus creating prerequisites for studying new treatment approaches.

Keywords: maxillofacial surgery, microflora, odontogenic phlegmons, pyo-inflammatory

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12 Transcriptional Response of Honey Bee to Differential Nutritional Status and Nosema Infection

Authors: Farida Azzouz-Olden, Arthur G. Hunt, Gloria Degrandi-Hoffman

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Bees are confronting several environmental challenges, including the intermingled effects of malnutrition and disease. Intuitively, pollen is the healthiest nutritional choice; however, commercial substitutes, such as BeePro and MegaBee, are widely used. Herein we examined how feeding natural and artificial diets shapes transcription in the abdomen of the honey bee, and how transcription shifts in combination with Nosema parasitism. Gene ontology enrichment revealed that, compared with poor diet (carbohydrates (C)), bees fed pollen (P > C), BeePro (B > C), and MegaBee (M > C) showed a broad upregulation of metabolic processes, especially lipids; however, pollen feeding promoted more functions and superior proteolysis. The superiority of the pollen diet was also evident through the remarkable overexpression of vitellogenin in bees fed pollen instead of MegaBee or BeePro. Upregulation of bioprocesses under carbohydrates feeding compared to pollen (C > P) provided a clear poor nutritional status, uncovering stark expression changes that were slight or absent relatively to BeePro (C > B) or MegaBee (C > M). Poor diet feeding (C > P) induced starvation response genes and hippo signaling pathway, while it repressed growth through different mechanisms. Carbohydrate feeding (C > P) also elicited ‘adult behavior’, and developmental processes suggesting transition to foraging. Finally, it altered the ‘circadian rhythm’, reflecting the role of this mechanism in the adaptation to nutritional stress in mammals. Nosema-infected bees fed pollen compared to carbohydrates (PN > CN) upheld certain bioprocesses of uninfected bees (P > C). Poor nutritional status was more apparent against pollen (CN > PN) than BeePro (CN > BN) or MegaBee (CN > MN). Nosema accentuated the effects of malnutrition since more starvation-response genes and stress response mechanisms were upregulated in CN > PN compared to C > P. The bioprocess ‘Macromolecular complex assembly’ was also enriched in CN > PN, and involved genes associated with human HIV and/or influenza, thus providing potential candidates for bee-Nosema interactions. Finally, the enzyme Duox emerged as essential for guts defense in bees, similarly to Drosophila. These results provide evidence of the superior nutritional status of bees fed pollen instead of artificial substitutes in terms of overall health, even in the presence of a pathogen.

Keywords: honeybee, immunity, Nosema, nutrition, RNA-seq

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11 Bioactive Secondary Metabolites from Culturable Unusual Actinomycetes from Solomon Islands Marine Sediments: Isolation and Characterisation of Bioactive Compounds

Authors: Ahilya Singh, Brad Carte, Ramesh Subramani, William Aalbersberg

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A total of 37 actinomycete strains were purified from 25 Solomon Islands marine sediments using four different types of isolation media. Among them, 54% of the strains had obligate requirement of seawater for growth. The ethyl acetate extract of 100 ml fermentation product of each strain was screened for antimicrobial activity against multidrug resistant human pathogens and cytotoxic activity against brine shrimps. A total of 67% of the ethyl acetate extracts showed antimicrobial and/or cytotoxic activities. A strain F-1915 was selected for isolation and evaluation of bioactive compound(s) based on its bioactive properties and chemical profile analysis using the LC-MS. The strain F-1915 was identified to have 96% sequence similarity to Streptomyces violaceusniger on the basis of 16S rDNA sequences using BLAST analysis. The 16S rDNA revealed that the strain F-1915 is a new member of MAR4 clade of actinomycetes. The MAR4 clade is an interesting clade of actinomycetes known for the production of pharmaceutically important hybrid isoprenoid compounds. The ethyl acetate extract of the fermentation product of this strain was purified by silica gel column chromatography and afforded the isolation of one bioactive pure compound. Based on the 1D and 2D NMR spectral data of compound 1 it was identified as a new mono-brominated phenazinone, Marinophenazimycin A, a structure which has already been studied by external collaborators at Scripps Institution of Oceanography but is yet to be published. Compound 1 displayed significant antimicrobial activity against drug resistant human pathogens. The minimum inhibitory concentration (MIC) of compound 1 was against Methicillin Resistant Staphylococcus aureus (MRSA) was about 1.9 μg/ml and MIC recorded against Amphotericin Resistant Candida albicans (ARCA) was about 0.24 μg/ml. The bioactivity of compound 1 against ARCA was found to be better than the standard antifungal agent amphotericin B. Compound 1 however did not show any cytotoxic activity against brine shrimps.

Keywords: actinomycetes, antimicrobial activity, brominated phenazine, MAR4 clade, marine natural products, multidrug resistent, 1D and 2D NMR

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10 In vitro and in vivo Infectivity of Coxiella burnetii Strains from French Livestock

Authors: Joulié Aurélien, Jourdain Elsa, Bailly Xavier, Gasqui Patrick, Yang Elise, Leblond Agnès, Rousset Elodie, Sidi-Boumedine Karim

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Q fever is a worldwide zoonosis caused by the gram-negative obligate intracellular bacterium Coxiella burnetii. Following the recent outbreaks in the Netherlands, a hyper virulent clone was found to be the cause of severe human cases of Q fever. In livestock, Q fever clinical manifestations are mainly abortions. Although the abortion rates differ between ruminant species, C. burnetii’s virulence remains understudied, especially in enzootic areas. In this study, the infectious potential of three C. burnetii isolates collected from French farms of small ruminants were compared to the reference strain Nine Mile (in phase II and in an intermediate phase) using an in vivo (CD1 mice) model. Mice were challenged with 105 live bacteria discriminated by propidium monoazide-qPCR targeting the icd-gene. After footpad inoculation, spleen and popliteal lymph node were harvested at 10 days post-inoculation (p.i). The strain invasiveness in spleen and popliteal nodes was assessed by qPCR assays targeting the icd-gene. Preliminary results showed that the avirulent strains (in phase 2) failed to pass the popliteal barrier and then to colonize the spleen. This model allowed a significant differentiation between strain’s invasiveness on biological host and therefore identifying distinct virulence profiles. In view of these results, we plan to go further by testing fifteen additional C. burnetii isolates from French farms of sheep, goat and cattle by using the above-mentioned in vivo model. All 15 strains display distant MLVA (multiple-locus variable-number of tandem repeat analysis) genotypic profiles. Five of the fifteen isolates will bee also tested in vitro on ovine and bovine macrophage cells. Cells and supernatants will be harvested at day1, day2, day3 and day6 p.i to assess in vitro multiplication kinetics of strains. In conclusion, our findings might help the implementation of surveillance of virulent strains and ultimately allow adapting prophylaxis measures in livestock farms.

Keywords: Q fever, invasiveness, ruminant, virulence

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9 Effect of Inoculum Ratio on Dark Fermentative Hydrogen Production

Authors: Zeynep Yilmazer Hitit, Patrick C. Hallenbeck

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Fuel reserve requirements due to depletion of fossil fuels have increased interest in biohydrogen since the 1990’s. In fermentative hydrogen production, pure, mixed, and co-cultures can be used to produce hydrogen. Several previous studies have evaluated hydrogen production by pure cultures of Clostridium butyricum or Enterobacter aerogenes. Evaluating hydrogen production by co-culture of these microorganisms is an interestıng approach since E. aerogenes is a facultative microorganism with resistance to oxygen in contrast to the strict anaerobe C. butyricum, and therefore has the ability to maintain anaerobic conditions. It was found that using co-cultures of facultative E. aerogenes (as a reducing agent and H2 producer) and the obligate anaerobe C. butyricum for producing hydrogen increases the yield of hydrogen by about 50% compared to C. butyricum by itself. Also, using different types of microorganisms for hydrogen production eliminates the need to use expensive reducing agents. C. butyricum strain pre-cultured anaerobically at 37 0C for 15h by inoculating 100 mL of GP medium (pH 6.8) consisting of 1% glucose, 2% polypeptone, 0.2% KH2PO4, 0.05% yeast extract, 0.05% MgSO4. 7H2O and E. aerogenes strain was pre-cultured aerobically at 30 0C, 150 rpm for 9 h by inoculating 100 mL of TGY medium (pH 6.8), consisting of 0.1% glucose, 0.5% tryptone, 0.1% K2HPO4, 0.5% yeast extract. All duplicate batch experiments were conducted in 100 mL bottles with different inoculum ratios of Clostridium butyricum and Enterobater aerogenes (C:E) using 5x diluted rich media (GP) consisting of 2 g/L glucose, 4g/L polypeptone, 0.4 g/L KH2PO4, 0.1 g/L yeast extract, 0.1 MgSO4.7H2O. The range of inoculum ratio of C. butyricum to E. aerogenes were 2:1,4:1,8:1, 1:2,1:4, 1:8, 1:0, 0:1. Using glucose as a carbon source aided in the observation of microbial behavior as well as making the effect of inoculum ratio more evident. Nearly all the glucose in the medium was used to produce hydrogen, except at a 1:0 ratio of inoculum (i.e. containing only C. butyricum). Low glucose consumption leads to a higher hydrogen yield due to cumulative hydrogen production and consumption of glucose, but not as much as C:E, 8:1. The lowest hydrogen yield was achieved in 1:8 inoculum ratio of C:E, 71.9 mL, 1.007±0.01 mol H2/mol glucose and the highest cumulative hydrogen, hydrogen yield and dry cell weight were achieved in 8:1 inoculum ratio of C:E, 117.4 mL, 2.035±0.082 mol H2/mol glucose, 0.4 g/L respectively. In this study effect of inoculum ratio on dark fermentative biohydrogen production using C. butyricum and E. aerogenes was investigated. The maximum hydrogen yield of 2.035mol H2/mol glucose was obtained using 2g/L glucose, an initial pH of 6 and an inoculum ratio of C. butyricum to E. aerogenes of 8:1. Results showed that inoculum ratio is an important parameter on hydrogen production due to competition between the two microorganisms in using substrate for growth and production of by-products. The results presented here could be of great significance for further waste management studies using co-culture hydrogen production.

Keywords: biohydrogen, Clostridium butyricum, dark fermentation, Enterobacter aerogenes, inoculum ratio in biohydrogen production

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8 Association of Ovine Lymphocyte Antigen (OLA) with the Parasitic Infestation in Kashmiri Sheep Breeds

Authors: S. A. Bhat, Ahmad Arif, Muneeb U. Rehman, Manzoor R Mir, S. Bilal, Ishraq Hussain, H. M Khan, S. Shanaz, M. I Mir, Sabhiya Majid

Abstract:

Background: Geologically Climatic conditions of the state range from sub-tropical (Jammu), temperate (Kashmir) to cold artic (Ladakh) zones, which exerts significant influence on its agro-climatic conditions. Gastrointestinal parasitism is a major problem in sheep production worldwide. Materials and Methods: The present study was to evaluate the resistance status of sheep breeds reared in Kashmir Valley for natural resistance against Haemonchus contortus by natural pasture challenge infection. Ten microsatellite markers were used in the study for evaluation of association of Ovar-MHC with parasitic resistance in association with biochemical and parasitological parameters. Following deworming, 500 animals were subjected to selected contaminated pastures in a vicinity of the livestock farms of SKUAST-K and Sheep Husbandry Kashmir. For each animal about 10-15 ml blood was collected aseptically for molecular and biochemical analysis. Weekly fecal samples (3g) were taken, directly from the rectum of all experimental animals and examined for Fecal egg count (FEC) with modified McMaster technique. Packed cell volume (PCV) was determined within 2-5 h of blood collection, all the biochemical parameters were determined in serum by semi automated analyzer. DNA was extracted from all the blood samples with phenol-chloroform method. Microsatellite analysis was done by denaturing sequencing gel electrophoresis Results: Overall sheep from Bakerwal breed followed by Corriediale breed performed relatively better in the trial; however difference between breeds remained low. Both significant (P<0.05) and non-significant differences with respect to resistance against haemonchosis were noted at different intervals in all the parameters.. All the animals were typed for the microsatellites INRA132, OarCP73, DRB1 (U0022), OLA-DQA2, BM1818, TFAP2A, HH56, BM1815, IL-3 and BM-1258. An association study including the effect of FEC, PCV, TSP, SA, LW, and the number of alleles within each marker was done. All microsatellite markers showed degree of heterozygosity of 0.72, 0.72, 0.75, 0.62, 0.84, 0.69, 0.66, 0.65, 0.73 and 0.68 respectively. Significant association between alleles and the parameters measured were only found for the OarCP73, OLA-DQA2 and BM1815 microsatellite marker. Standard alleles of the above markers showed significant effect on the TP, SA and body weight. The three sheep breeds included in the study responded differently to the nematode infection, which may be attributed to their differences in their natural resistance against nematodes. Conclusion: Our data confirms that some markers (OarCP73, OLA-DQA2 and BM1815) within Ovar-MHC are associated with phenotypic parameters of resistance and suggest superiority of Bakerwal sheep breed in natural resistance against Haemonchus contortus.

Keywords: Ovar-Mhc, ovine leukocyte antigen (OLA), sheep, parasitic resistance, Haemonchus contortus, phenotypic & genotypic markers

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7 Improving the Biocontrol of the Argentine Stem Weevil; Using the Parasitic Wasp Microctonus hyperodae

Authors: John G. Skelly, Peter K. Dearden, Thomas W. R. Harrop, Sarah N. Inwood, Joseph Guhlin

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The Argentine stem weevil (ASW; L. bonariensis) is an economically important pasture pest in New Zealand, which causes about $200 million of damage per annum. Microctonus hyperodae (Mh), a parasite of the ASW in its natural range in South America, was introduced into New Zealand to curb the pasture damage caused by the ASW. Mh is an endoparasitic wasp that lays its eggs in the ASW halting its reproduction. Mh was initially successful at preventing ASW proliferation and reducing pasture damage. The effectiveness of Mh has since declined due to decreased parasitism rates and has resulted in increased pasture damage. Although the mechanism through which ASW has developed resistance to Mh has not been discovered, it has been proposed to be due to the different reproductive modes used by Mh and the ASW in New Zealand. The ASW reproduces sexually, whereas Mh reproduces asexually, which has been hypothesised to have allowed the ASW to ‘out evolve’ Mh. Other species within the Microctonus genus reproduce both sexually and asexually. Strains of Microctonus aethiopoides (Ma), a species closely related to Mh, reproduce either by sexual or asexual reproduction. Comparing the genomes of sexual and asexual Microctonus may allow for the identification of the mechanism of asexual reproduction and other characteristics that may improve Mh as a biocontrol agent. The genomes of Mh and three strains of Ma, two of which reproduce sexually and one reproduces asexually, have been sequenced and annotated. The French (MaFR) and Moroccan (MaMO) reproduce sexually, whereas the Irish strain (MaIR) reproduces asexually. Like Mh, The Ma strains are also used as biocontrol agents, but for different weevil species. The genomes of Mh and MaIR were subsequently upgraded using Hi-C, resulting in a set of high quality, highly contiguous genomes. A subset of the genes involved in mitosis and meiosis, which have been identified though the use of Hidden Markov Models generated from genes involved in these processes in other Hymenoptera, have been catalogued in Mh and the strains of Ma. Meiosis and mitosis genes were broadly conserved in both sexual and asexual Microctonus species. This implies that either the asexual species have retained a subset of the molecular components required for sexual reproduction or that the molecular mechanisms of mitosis and meiosis are different or differently regulated in Microctonus to other insect species in which these mechanisms are more broadly characterised. Bioinformatic analysis of the chemoreceptor compliment in Microctonus has revealed some variation in the number of olfactory receptors, which may be related to host preference. Phylogenetic analysis of olfactory receptors highlights variation, which may be able to explain different host range preferences in the Microctonus. Hi-C clustering implies that Mh has 12 chromosomes, and MaIR has 8. Hence there may be variation in gene regulation between species. Genome alignment of Mh and MaIR implies that there may be large scale genome structural variation. Greater insight into the genetics of these agriculturally important group of parasitic wasps may be beneficial in restoring or maintaining their biocontrol efficacy.

Keywords: argentine stem weevil, asexual, genomics, Microctonus hyperodae

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6 Analysis of Aspergillus fumigatus IgG Serologic Cut-Off Values to Increase Diagnostic Specificity of Allergic Bronchopulmonary Aspergillosis

Authors: Sushmita Roy Chowdhury, Steve Holding, Sujoy Khan

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The immunogenic responses of the lung towards the fungus Aspergillus fumigatus may range from invasive aspergillosis in the immunocompromised, fungal ball or infection within a cavity in the lung in those with structural lung lesions, or allergic bronchopulmonary aspergillosis (ABPA). Patients with asthma or cystic fibrosis are particularly predisposed to ABPA. There are consensus guidelines that have established criteria for diagnosis of ABPA, but uncertainty remains on the serologic cut-off values that would increase the diagnostic specificity of ABPA. We retrospectively analyzed 80 patients with severe asthma and evidence of peripheral blood eosinophilia ( > 500) over the last 3 years who underwent all serologic tests to exclude ABPA. Total IgE, specific IgE and specific IgG levels against Aspergillus fumigatus were measured using ImmunoCAP Phadia-100 (Thermo Fisher Scientific, Sweden). The Modified ISHAM working group 2013 criteria (obligate criteria: asthma or cystic fibrosis, total IgE > 1000 IU/ml or > 417 kU/L and positive specific IgE Aspergillus fumigatus or skin test positivity; with ≥ 2 of peripheral eosinophilia, positive specific IgG Aspergillus fumigatus and consistent radiographic opacities) was used in the clinical workup for the final diagnosis of ABPA. Patients were divided into 3 groups - definite, possible, and no evidence of ABPA. Specific IgG Aspergillus fumigatus levels were not used to assign the patients into any of the groups. Of 80 patients (males 48, females 32; mean age 53.9 years ± SD 15.8) selected for the analysis, there were 30 patients who had positive specific IgE against Aspergillus fumigatus (37.5%). 13 patients fulfilled the Modified ISHAM working group 2013 criteria of ABPA (‘definite’), while 15 patients were ‘possible’ ABPA and 52 did not fulfill the criteria (not ABPA). As IgE levels were not normally distributed, median levels were used in the analysis. Median total IgE levels of patients with definite and possible ABPA were 2144 kU/L and 2597 kU/L respectively (non-significant), while median specific IgE Aspergillus fumigatus at 4.35 kUA/L and 1.47 kUA/L respectively were significantly different (comparison of standard deviations F-statistic 3.2267, significance level p=0.040). Mean levels of IgG anti-Aspergillus fumigatus in the three groups (definite, possible and no evidence of ABPA) were compared using ANOVA (Statgraphics Centurion Professional XV, Statpoint Inc). Mean levels of IgG anti-Aspergillus fumigatus (Gm3) in definite ABPA was 125.17 mgA/L ( ± SD 54.84, with 95%CI 92.03-158.32), while mean Gm3 levels in possible and no ABPA were 18.61 mgA/L and 30.05 mgA/L respectively. ANOVA showed a significant difference between the definite group and the other groups (p < 0.001). This was confirmed using multiple range tests (Fisher's least significant difference procedure). There was no significant difference between the possible ABPA and not ABPA groups (p > 0.05). The study showed that a sizeable proportion of patients with asthma are sensitized to Aspergillus fumigatus in this part of India. A higher cut-off value of Gm3 ≥ 80 mgA/L provides a higher serologic specificity towards definite ABPA. Long-term studies would provide us more information if those patients with 'possible' APBA and positive Gm3 later develop clear ABPA, and are different from the Gm3 negative group in this respect. Serologic testing with clear defined cut-offs are a valuable adjunct in the diagnosis of ABPA.

Keywords: allergic bronchopulmonary aspergillosis, Aspergillus fumigatus, asthma, IgE level

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