emergence of salmonella as a food borne pathogen,march 2015

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MICROBIOZ INDIA MICROBIOZ INDIA ISSUE. MARCH 2015. VOLUME .02 www.microbiozindia.com almonella EMERGENCE OF Salmonella Species AS A FOOD-BORNE PATHOGEN FREE SUBSCRIPTION Special Issue PUZZLE WORD INSIDE!! Microbioz recent research updates Open scholarship positions M J O U R N A L S CALL FOR PAPERS List of Winners of February Issue S An Interview with Pf. Aussielita L. Lit, PHILIPPINES

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Emergence of Salmonella species,as a food borne PathogenCover Story Microbioz India going to launch first issue of our 2nd volume in the month of March 2015,I would like to thanks to all of our respected team members, supporters who always appreciating me well. Dear readers Microbioz India, March issue strictly focuses of “Food Borne Pathogens” a special issue on world water day: 22nd March 2015, the cover story of this issue is entitled: “Emergence of Salmonella species as a food-borne pathogen”. The cover story is cover by our team member Ms. Sivashankari Ramamoorthi, from University of Malaya, Malaysia. As cover story depicts that Emergence of Salmonella species as a food borne pathogen here you can collect number of specific information about different aspects of Salmonella Species in food borne infections. Several factors will influence the stability of Salmonella spp growth in foods. One of the factors is known as intrinsic factors such as pH, water activity (aw) and other physicochemical properties of food such as processing which includes cooking temperature. The other factor is known as extrinsic factor includes environmental conditions, food storage and temperatures. These factors are playing an important role in Salmonella growth and this is why the outbreak of foodborne pathogens usually involves specific types of foods (Jeníková et al., 2010).Salmonella plays an important role economically. Despite of its harmful effects,these pathogens provides novel strategies for cancer immunotherapy. Salmonella typhimurium had shown some promises for development of microbial-based tumor therapies via genetic engineering. Lipid A negative strain of S. typhimurium successfully completed the phase I clinical trials and harmless at doses up to 109 CFU per m2 body surface area (Le Negrate et al., 2008).Apart from magazines also has a huge collection of different research news and information collected from worldwide sources.An Interview with Prof. Aussielita L. Lit from Philippines As we did in earlier issue in this month our team representative from Phillipines Ms.Rodel Estadillo Alo perform an interview with Prof.Aussielita L. Lit Philippines. Microbioz India,wish Prof.Great Success in her future.Recent Research News InformationMicrobioz india,march issue contains number of recent research news information collected from world wide sources.Microbioz India,Recent open Scholarship PositionsAs per demand of our academic reader’s magazines also have collections of different open scholarship positions for all those students having interest in pursuing higher education in Microbiology from different reputed University of world.Microbioz India,Cross Word March 2015 and winners of Last editionsAnd in last how we can forget to announce the list of winners of cross word winners of February 2015 editions

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  • MICROBIOZ INDIA

    MICROBIOZ INDIA

    ISSUE. MARCH 2015. VOLUME .02

    www.microbiozindia.com

    almonella EMERGENCE OF Salmonella Species AS A FOOD-BORNE PATHOGEN FR

    EE S

    UBS

    CRIP

    TIO

    N

    Special Issue

    PPUUZZZZLLEE WWOORRDD

    IINNSSIIDDEE!! !!

    Microbioz recent research updates

    Open scholarship positions M J O U R N A L S

    CALL FOR PAPERS

    List of Winners of February Issue

    S An Interview with Pf. Aussielita L. Lit, PHILIPPINES

  • MICROBIOZ INDIA

    MONTHLY MICROBIOLOGY MAGAZINES & INTERNATIONAL JOURNALS

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    A peer reviewed specific International Journal of Microbiology

    www.facebook.com/MICROBIOZINDIA www.twitter.com/MICROBIOZINDIA www.microbiozindia.com

    Microbioz Journals call for Research Article/Review article/Short

    Communications/Case Studies/letter to editor/Scientific Manuscripts for

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  • Contents

    Disease surveillance reports frequently identify poultry, meat and milk products as the main vehicles in Salmonellosis outbreaks. However, recent years of food borne illness outbreaks have been linked to greater consumption of fresh fruits and vegetables (CDC, 2012).

    Microbioz India, Cover Story, Emergence of Salmonella Spp, as a food borne Pathogen

    List of Crossword winners of February Edition

    An Interview with Prof. Aussielita L. Lit Philippines

    Microbioz India, Recent Research News, Collected from Worldwide sources.

    Microbioz India,Puzzle Word Game of March 2015

    Recent Open Scholarships Positions

    36 27

    36 12

    29 06

  • MICROBIOZ INDIA

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    Editorial Lines DDear readers,, Microbioz India going to launch first issue of our 2nd volume in the month of March 2015,I would like to thanks to all of our respected team members, supporters who always appreciating me well. Dear readers Microbioz India, March issue strictly focused of Food Borne Pathogens a special issue on world water day: 22nd March 2015, the cover story of this issue is entitled: Emergence of Salmonella species as a food-borne pathogen. The cover story is covered by our team member Ms. Sivashankari Ramamoorthi, from University of Malaya, Malaysia. As cover story depicts that Emergence of Salmonella species as a food borne pathogen here you can collect number of specific information about different aspects of Salmonella Species in food borne infections. Several factors will influence the stability of Salmonella spp growth in foods. One of the factors is known as intrinsic factors such as pH, water activity (aw) and other physicochemical properties of food such as processing which includes cooking temperature. The other factor is known as extrinsic factor includes environmental conditions, food storage and temperatures. These factors are playing an important role in Salmonella growth and this is why the outbreak of foodborne pathogens usually involves specific types of foods (Jenkov et al., 2010).

    Salmonella plays an important role economically. Despite of its harmful effects, these pathogens provides novel strategies for cancer immunotherapy. Salmonella typhimurium had shown some promises for development of microbial-based tumor therapies via genetic engineering. Lipid A negative strain of S. typhimurium successfully completed the phase I clinical trials and harmless at doses up to 109 CFU per m2 body surface area (Le Negrate et al., 2008).Apart from magazines also has a huge collection of different research news and informations collected from worldwide sources. As we did in earlier issue in this month our team representative from Phillipines Ms.Rodel Estadillo Alo perform an interview with Prof.Aussielita L. Lit Philippines. As per demand of our academic readers magazines also have collections of different open scholarship positions for all those students having interest in pursuing higher education in Microbiology from different reputed University of world. And in last how we can forget to announce the list of winners of cross word winners of February 2015 editions.

    Thanking you

    Kumaar Jeetendra

    Chief Editor, Microbioz India

    (Magazines & International Journals)

    MICROBIOZ INDIA MARCH 2015 www.microbiozindia.com 4

  • almonella S EMERGENCE OF Salmonella Species AS A FOOD-BORNE PATHOGEN Consumption of contaminated street-vended foods that have high levels of coliform bacteria and the presence of pathogenic bacteria, such as Escherichia coli, Salmonella spp, Staphylococcus aureus, Bacillus cereus, Clostridium perfringens, and Vibrio choleraresults in severe foodborne illnesses. People who patronize the vendors of RTE street food are putting their health at risk (Manguiat & Fang, 2013).

    In recent decades, public health promotion of healthier lifestyles has led to increased demand for fresh produce which is one of the types of RTE food. However, fruits and vegetables, and in particular leafy greens that are consumed raw, are increasingly being recognized as important vehicles for transmission of human pathogens (Berger et al, 2010). Vegetables including cauliflower, lettuce and spinach have been reported as vehicles of Salmonella spp (Quiroz-Santiago et al, 2009). Salmonellosis outbreaks in USA have been linked to fresh tomatoes where the cross contamination during post-harvest washing of tomatoes caused the spreading of Salmonella spp from contaminated tomatoes to non-contaminated tomatoes (Keller, 2009).

    Cover Story Cover by: Sivashankari Ramamoorthi

    -University of Malaya, Malaysia

  • SS

    almonella spp is not a new species in microbiology field. It is an old microorganism which had a huge impact on history. Researchers from microbiology field have stated that the death of Alexander the Great could be caused by Salmonella spp in 323 BC (Medicine Net, 2012). Besides, Salmonella spp is also regarded as one of the early contagious microorganism to be identified in microbiology field that can cause disease (Mastroeni & Maskell, 2006).

    Salmon and Theobald Smith were the early scientists who successfully isolated disease causing Salmonella spp from the infected pigs in the year of 1885. They have isolated Salmonella enterica serovar Typhi from those infected pigs. However, the genus of bacteria is named as Salmonella in honor to Dr. Salmon and not after Theobald Smith because Dr.Salmon was the administrator of United States of Department of Agriculture (USDA) research program at that time (Clark, 1959). Even though Salmonella spp has been discovered on 1885, the infections caused by Salmonella spp is only been started to be diagnosed on 1986 (Medicine Net, 2012).

    Nomenclature and Taxonomy of the genus Salmonella

    In the classification of Salmonella, two terms are generally used in frequent which are serotype and serovar. However, serovar term is preferred over the term serotype as being recommended in Rules of the Bacteriological Code established by Judicial Commission of the International Committee on the Systematics of Prokaryotes. Therefore, in the Kauffmann- White scheme the term serovar is used instead of serotype (Grimont & Weill, 2007).

    Salmonella is a group of bacteria that are being distinguished by the serovars since they share great genetic traits in similar. The serotyping is done based on antigenic interactions of Salmonella with antibodies. The different antibody-antigen reactions aids in the determination of serotypes of Salmonella. The classification system of vast number of Salmonellae based on serovars that is in todays use is actually an accumulated results of many studies on the interaction of antigens on Salmonella and antibodies in previous years as being suggested by Kauffmann and White.

    The classification of Salmonella by antigen-antibody is a very systemic process in which all the antigenic formulae of identified Salmonella serovars are updated neatly in a document which is more commonly known as Kauffmann-White scheme (Popoff et al.,2004). The World Health Organization Collaborating Centre for Reference and Research in Salmonella at the Pasteur Institute, Paris, France (WHO Collaborating Centre) is the organization that take charge of compiling and update the scheme whenever a new serovar is being recognized.

    Cover Story

    History Salmonella species

    Theobald Smith

    MICROBIOZ INDIA MARCH 2015 www.microbiozindia.com 6

  • Annually, the newly identified Salmonella serovars will be reported by Popoff et al to the Research in Microbiology as to enable the updating system to work in a systematic manner (Popoff et al., 2004). As in the latest report on the serovars of Salmonella, the genus of this foodborne pathogen currently containing 2,587 serovars (Fashae et al., 2010).The nomenclature of Salmonella is a complicated system and always evolving from time to time. Recommendation from the World Health Organization Collaborating Centre (WHO Collaborating Centre) is now widely used in the Centers for Disease Control and Prevention (CDC) for the nomenclature system of genus Salmonella.The latest nomenclature system had suggested that the genus Salmonella is only made up of two types of species which are Salmonella enterica (S.enterica) and Salmonella bongori (S.bongori) (Grimont & Weill, 2007). However, a new species is identified and named as Salmonella subterranean on 18th March 2005 was officially approved by Judicial Commission (Shelobolina et al., 2004), thus CDC might include this new species in the nomenclature system in future. On the other hand, a more recent unpublished data suggest that this newly identified organism does not actually belong in the genus Salmonella (Grimont & Weill, 2007). Therefore this issue is still on debate whether the new species can or cannot be included in the nomenclature system for genus Salmonella.

    The number of subspecies in S.enterica and S.bongori varies. As for S.enterica there are total of six subspecies as tabulated in the Table :1 (Brenner et al., 2000).

    Subspecies of S.enterica

    The serovars in subspecies I is assigned names as an indicative of the diseases they are associated with and their origins. The origins being mentioned here is both the geographic origin and their usual habitats.The subspecies enterica are also regarded as the most common serovar that cause infections in humans and food animals. On the other hand, other subspecies which are II , IIIa, IIIb, IV,VI and all the subspecies listed under S.bongori , the antigenic formulae are determined as being recommended in Kauffmann-White scheme. In additional, some of the subspecies are already assigned with names before 1966, therefore their names are retained and cited under subspecies I.In contrast, the serovars of the other subspecies are mostly prevalent in poikilothermic (cold-blooded) animals and in the environment (Popoff et al., 2004).

    Characteristics of Salmonella Salmonella spp is a dangerous foodborne pathogen which can contaminate the food items either before processing, during

    processing or after processing (Cabedo et al., 2008). Salmonella spp is a non- spore forming bacterium which stains red under light microscope and it is rod shaped. Besides, it is predominantly motile enterobacteria that has diameters ranging from 0.7 to 1.5 m, and length from 2 to 5 m, with flagella that grade in all directions (Pui et al., 2011).Furthermore, it is a facultative anaerobic, and chemoorganotrophic organism which means it has the ability to degrade organic material to derive energy for survival (Neidhardt et al., 1996). Biochemically, Salmonella spp is catalase positive, oxidase negative and produce hydrogen sulphide gas (Wray & Davies, 2003).

    Types Salmonella enterica subspecies

    I Enterica

    II Salamae

    III a Arizonae

    III b Diarizonae

    IV Houtenae

    VI Indica

    Cover Story

    MICROBIOZ INDIA MARCH 2015 www.microbiozindia.com 7

  • Salmonella spp is the most worrisome of the pathogenic microorganisms found in minimally processed fresh produce (Beuchat, 1996). Salmonella has been isolated from fruits and vegetables such as cantaloupes, melons, tomatoes, lettuce and especially alfalfa sprouts.

    Several factors will influence the stability of Salmonella spp growth in foods. One of the factors is known as intrinsic factors such as pH, water activity (aw) and other physicochemical properties of food such as processing which includes cooking temperature. The other factor is known as extrinsic factor includes environmental conditions, food storage and temperatures. These factors are playing an important role in Salmonella growth and this is why the outbreak of foodborne pathogens usually involves specific types of foods (Jenkov et al., 2010).

    Salmonella plays an important role economically. Despite of its harmful effects, these pathogens provides novel strategies for cancer immunotherapy. Salmonellatyphimurium had shown some promises for development of microbial-based tumor therapies via genetic engineering. Lipid A negative strain of S.typhimurium successfully completed the phase I clinical trials and harmless at doses up to 109 CFU per m2 body surface area (Le Negrate et al., 2008).

    Salmonellosis in Food Salmonellosis is a disease caused by any Salmonella serovars. Non typhoidal

    salmonellosis is infection caused by other serovars of Salmonella than Salmonella Typhi and SalmonellaParatyphi. Salmonellosis is commonly related with S. enterica that comprises of 6 subspecies which are potentially pathogenic with more than 2000 serovars (Black et al., 1960).

    Salmonella enterica subspecies enterica serovar Enteritidis (S. Enteritidis) is an important causative agent of non-typhoidal salmonellosis in human populations. The emergence and epidemic spread of this pathogen began in mid-80's (Velge et al., 2005). To date, S. Enteritidis outbreaks are still a major public health concern in both developed and developing countries (Betancor et al., 2010 and Solhan et al., 2011).

    The hosts for Salmonella ranging from human and other warm and cold blooded animals results in rapid transmission of Salmonella from one host to another without considering species barrier. The transmissions occur when an individual had consumed Salmonella spp contaminated food such as chicken, milk, vegetables (Colville & Berryhill, 2007).

    The dynamics of Salmonella infection is variable and may also be affected by human lifestyle and behavior, changes in industry, technology, commerce and travel (Foley et al., 2008). Salmonella serovars are widespread in nature and can be found in the intestinal tract of all animals species, both domestic and wild which result in a variety of Salmonella infection sources (Callaway et al., 2005).

    The clinical symptoms of salmonellosis are commonly diarrhea, fever, vomiting, stomach cramps and nausea. Besides, severe cases such as dehydration might be observed in elderly people and infants. Enteric fever also may result from salmonellosis (Feasey & Gordon, 2014).

    Salmonellosis in Ready to Eat food RTE food are defined as food that can be consumed immediately at the point of sale without further preparation or treatment. It

    could be raw, partially or fully cooked, and hot, chilled or frozen (Ng, et al., 2013). Since RTE food are edible without additional treatment, risks of foodborne outbreaks are high if it is improperly handled and it is highly subject to bacterial foodborne outbreaks (Fajardo et al.,2012).Various foodborne pathogens associated with RTE food have been found to contribute to foodborne outbreaks (Castro-Rosas et al., 2012 and Seow et al., 2012). Methods of storage, processing, handling, and display can affect the levels of microorganisms in ready-to-eat food (Christison et al., 2008 and Fang, et. al, 2013).

    Foodborne illness outbreaks are occasionally associated with RTE sandwiches and other heat and eat multi-component RTE products (Christopher& Sommers, 2006). RTE food are food that will not be cooked or reheated before serving and consumed at sale point (Kotzekidou, 2013). RTE food include salads, cooked meats, desserts, sandwiches, cheese and foods that have been cooked advance to serve cold.

    Cover Story

    MICROBIOZ INDIA MARCH 2015 www.microbiozindia.com 8

  • These type of food should be consumed within 4 hours to reduce the prevalence of Salmonella spp and other food borne pathogens (Fang et al. 2003).RTE food does not undergo any treatment to ensure its safety before consumption, and therefore risk of foodborne disease must be considered if these pathogens are present in the food (Cabedo et al, 2008).

    Consumption of contaminated street-vended foods that have high levels of coliform bacteria and the presence of pathogenic bacteria, such as Escherichia coli, Salmonella spp,Staphylococcus aureus, Bacillus cereus, Clostridium perfringens, and Vibrio choleraresults in severe foodborne illnesses. People who patronize the vendors of RTE street food are putting their health at risk (Manguiat & Fang, 2013).

    In recent decades, public health promotion of healthier lifestyles has led to increased demand for fresh produce which is one of the types of RTE food. However, fruits and vegetables, and in particular leafy greens that are consumed raw, are increasingly being recognized as important vehicles for transmission of human pathogens (Berger et al, 2010). Vegetables including cauliflower, lettuce and spinach have been reported as vehicles of Salmonella spp (Quiroz-Santiago et al, 2009). Salmonellosis outbreaks in USA have been linked to fresh tomatoes where the cross contamination during post-harvest washing of tomatoes caused the spreading of Salmonella spp from contaminated tomatoes to non- contaminated tomatoes (Keller, 2009). Salmonella spp is the most worrisome of the pathogenic microorganisms found in minimally processed fresh produce (Beuchat, 1996). Salmonella has been isolated from fruits and vegetables such as cantaloupes, melons, tomatoes, lettuce and especially alfalfa sprouts (Berger et al, 2010). These products may be contaminated by several routes and contamination can be introduced through the roots, stem scars, and leaves of several different vegetables and fruits (Hedberg et al, 1999). It is thought that the source of contamination of most fresh produce is through untreated irrigation waters (Heard, 2002 and Jablasone et al, 2005). Therefore, consumers need to thoroughly wash all fresh food before consumption to reduce the risk of illness from fruits and vegetables.

    Disease surveillance reports frequently identify poultry, meat and milk products as the main vehicles in salmonellosis outbreaks. However, recent years of food borne illness outbreaks have been linked to greater consumption of fresh fruits and vegetables (CDC, 2012). The proportion of fresh produce samples that yielded Salmonella is ranged from 0.1% to 2.3% with pre-cut products having some of the highest proportion being contaminated (Berger et al, 2010).

    Microbiological methods to determine the prevalence of Salmonella species in ready to eat food

    Microbiological methods to determine the presence of Salmonella spp ranged from conventional methods to rapid methods (Blood, 1985).The conventional method is the gold standard to detect Salmonella from food which involves non selective pre-enrichment, followed by a selective enrichment step, isolation on selective agar media, purification on non-selective media and a preliminary biochemical confirmation (van der Zee, 1994). Pre-enrichment in Buffered Peptone Water (BPW) is vital to recover sub lethally injured Salmonella cells in the food samples (Myint et al., 2006).

    Cover Story

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  • Selective medium contains selective agents that only select targeted organisms and in selective plating, the main types of plating media used are Bismuth Sulphite agar, Hektoen Enteric agar and Xylose-lysine-Desoxycholate (XLD) agar (Cudjoe et al., 1994). The presumptive Salmonella colony morphology on XLD agar is colorless or yellow margin usually with a black center (Mansilha et al., 2010). A new selective media CHROMagar Salmonella Medium is proposed recently which incorporated with chromogenic substances to create a better differentiation of targeted organisms (Church et al., 2010).Biochemical tests aids in the determination of Salmonella spp presumptively. The table 01 shows characteristics and biochemical test results for Salmonella enterica subspecies enterica (Todar, 2009).

    Table 2: Characteristics and biochemical test results for Salmonella enterica subspecies enterica (Todar, 2009).

    Test Result

    Lactose Negative

    Glucose Acid and Gas

    Mannitol Acid and Gas

    Maltose Acid and Gas

    Sorbitol Acid and Gas

    ONPG test Negative

    Indole Negative

    Methyl red Positive

    Voges-Proskauer Negative

    Citrate Positive

    Lysine decarboxylase Positive

    Urease Negative

    Ornithine decarboxylase Positive

    Thiosulfate H2S produced

    KCN Does not grow

    Phenylalanine Negative

    Tryptophan deaminase Negative

    Gelatin hydrolysis Negative

    A rapid method can be either assay that gives instant or real time results, or it also can be a simple modification of a procedure that reduces the assay time (Yeni et al., 2014). These rapid methods not only deals with the early detection and enumeration of microorganisms, but also with the characterization of isolates by use of microbiological, chemical, biochemical, biophysical, molecular biological, immunological and serological methods (Kado & Liu, 1981).

    Cover Story

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  • Progress in molecular techniques has created opportunities in developing new methods. Several phenotypic, genotypic and molecular techniques such as bio-typing, phage typing, polymerase chain reaction (PCR), pulsed-field gel electrophoresis (PFGE) and nucleic acid hybridization have been developed for Salmonella identification and serotyping (Lagatolla et al., 1996 ).Among these, PCR has successfully been applied for detection of pathogenic bacteria using a member of largest sequences (Malkawi et al., 2008).

    Besides, the gold standard for subtyping S. Enteritidis is pulsed-field gel electrophoresis (PFGE), with standardized protocol developed for inter-laboratories comparison (Gerner-Smidt et al., 2006). However, PFGE is still insufficient to discriminate S. Enteritidis in Malaysia due to their clonal nature (Thong et al., 1995). Since 2000, multiple-locus variable-number tandem repeat analysis (MLVA) has been proposed for bacterial subtyping (van Belkum, 2007). MLVA subtypes a bacterial species based on the detection of variation in the variable-number tandem repeat (VNTR) found in the microbial genome at various regions (van Belkum, 2007). To date, several MLVA schemes with a number of VNTR loci in common have been developed for subtyping S.Enteritidis (Malorny et al., 2008).

    Besides, the use of selective and differential plating media is a simple method for the isolation of Salmonella spp. A wide variety of selective and differential media has been developed for this purpose, including Xylose Lysine Desoxycholate agar (XLD), Hektoen enteric (HE) agar, and bismuth sulfite (BS) agar (Cooke et al., 1999).

    XLD and HE agar are the most popular media for isolating Salmonella spp and their differentiation abilities rely on characteristics of Salmonella, such as hydrogen sulfide production and the non-fermentation of lactose (Dusch & Altwegg, 1995). However, these characteristics are shared with other microorganisms, such as Proteus and Citrobacter (Eigner et al., 2001). Thus, numerous false-positive results are observed on these media which require further confirmation testing, a time-consuming and labor-intensive activity (Perez et al., 2003). BS agar is the medium of choice for the isolation of Salmonella enterica serovar Typhi, and it is used for the isolation of atypical salmonellae, such as those which ferment lactose (Hu et al., 1997). However, BS agar has several disadvantages, such as low sensitivity and long incubation time for development of the characteristic colony morphology (Park et al., 2012).

    Several chromogenic media have been developed to increase the specificity of conventional selective and differential media for the detection of Salmonella spp (Schnenbrcher et al., 2008). These media incorporate chromogenic substrates that are metabolized by Salmonella spp (Ruiz et al., 1996). Although chromogenic media have higher specificities than conventional media, some of them have a low sensitivity which results in more false negatives observed on these media (Ruiz et al., 1996). Besides, chromogenic media are relatively expensive, making them less appropriate for routine laboratory use (Perry & Freydiere, 2007).

    Recognizing the limits of currently used selective and differential media, it is desirable to improve the specificity and the sensitivity of the medium while maintaining cost-effectiveness. Particularly, it is desirable to differentiate Salmonella spp. from Proteus spp., as well as from Citrobacter spp (Eigner et al., 2001). Hydrogen sulfide production depends on several factors, such as the sulfide production rate of the microorganisms, the oxygen concentration in the colony, pH, and the iron concentration in the medium Acid production by microorganisms in consequence of carbohydrate fermentation could inhibit hydrogen sulfide production (Park, Ryu, & Kang, 2012).

    The traditional method for detecting this food-borne pathogen is time consuming, consisting of enrichment steps, plating on selective media and incubation for a period of time. Nowadays, molecular methods by PCR have been extensively used in various studies for example duplex real-time PCR (Rodriguez-Lazaro et al., 2004), multiplex PCR (Hamdi et al., 2007) and Most Probable Number PCR (Marian et al, 2012).

    It is worthy to note that the MPN-PCR method is an effective tool to simultaneously detect the occurrence of food-borne pathogens quantitatively and qualitatively. The estimation of Salmonella spp density present in food samples can be calculated based on the reference MPN table provided by the U.S. Food and Drug Administration 2009.The MPN count method is the best applicable method for the detection of low levels of microorganisms in food samples which is in the range of 10100 MPN/g (Martin et al., 2004). Of late, the MPN-PCR method has been widely used in studies to detect the presence of food-borne pathogens in various food types quantitatively and qualitatively, and it was also claimed to be more useful and effective for detecting food borne pathogens such as Salmonella spp, Listeria monocytogens than was plating on media (Marian et al, 2012). Thus, application of the MPN-PCR method in the present study was suggested for more accurate and reliable results.

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    February 24, 2015

    WWhen diagnosing a case of Ebola, time is of the essence. However, existing diagnostic tests take at least a day or two to yield results, preventing health care workers from quickly determining whether a patient needs immediate treatment and isolation.A new test from MIT researchers could change that: The device, a simple paper strip similar to a pregnancy test, can rapidly diagnose Ebola, as well as other viral hemorrhagic fevers such as yellow fever and dengue fever.

    As we saw with the recent Ebola outbreak, sometimes people present with symptoms and its not clear what they have, says Kimberly Hamad-Schifferli, a visiting scientist in MITs Department of Mechanical Engineering and a member of the technical staff at MITs Lincoln Laboratory. We wanted to come up with a rapid diagnostic that could differentiate between different diseases.Hamad-Schifferli and Lee Gehrke, the Hermann L.F. von Helmholtz Professor in MITs Institute for Medical Engineering and Science (IMES), are the senior authors of a paper describing the new device in the journal Lab on a Chip. The papers lead author is IMES postdoc Chun-Wan Yen, and other authors are graduate student Helena de Puig, IMES postdoc Justina Tam, IMES instructor Jose Gomez-Marquez, and visiting scientist Irene Bosch. Color-coded test

    Currently, the only way to diagnose Ebola is to send patient blood samples to a lab that can perform advanced techniques such as polymerase chain reaction (PCR), which can detect genetic material from the Ebola virus. This is very accurate but time-consuming, and some areas of Africa where Ebola and other fevers are endemic have limited access to this kind of technology.The new device relies on lateral flow technology, which is used in pregnancy tests and has recently been exploited for diagnosing strep throat and other bacterial infections. Until now, however, no one has applied a multiplexing approach, using multicolored nanoparticles, to simultaneously screen for multiple pathogens. For many hemorrhagic fever viruses, like West Nile and dengue and Ebola, and a lot of other ones in developing countries, like Argentine hemorrhagic fever and the Hantavirus diseases, there are just no rapid diagnostics at all, says Gehrke, who began working with Hamad-Schifferli four years ago to develop the new device. Unlike most existing paper diagnostics, which test for only one disease, the new MIT strips are color-coded so they can be used to distinguish among several diseases. To achieve that, the researchers used triangular nanoparticles, made of silver that can take on different colors depending on their size. The researchers created red, orange, and green nanoparticles and linked them to antibodies that recognize Ebola, dengue fever, and yellow fever. As a patients blood serum flows along the strip, any viral proteins that match the antibodies painted on the stripes will get caught, and those nanoparticles will become visible. This can be seen by the naked eye; for those who are colorblind, a cellphone camera could be used to distinguish the colors. When we run a patient sample through the strip, if you see an orange band you know they have yellow fever, if it shows up as a red band you know they have Ebola, and if it shows up green then we know that they have dengue, Hamad-Schifferli says. This process takes about 10 minutes, allowing health care workers to rapidly perform triage and determine if patients should be isolated, helping to prevent the disease from spreading further. Warren Chan, an associate professor at the University of Toronto Institute of Biomaterials and Biomedical Engineering, says he is impressed with the device because it not only offers faster diagnosis, but also requires smaller patient blood samples, as just one test strip can detect multiple diseases. Its a step up from what everyone else is doing, says Chan, who was not involved in the research. Theyre targeting diseases that are really relevant to whats going on in the world at this point, and have shown that they can detect them simultaneously. Faster triage the researchers envision their new device as a complement to existing diagnostic technologies, such as PCR.

    If youre in a situation in the field with no power and no special technologies, if you want to know if a patient has Ebola, this test can tell you very quickly that you might not want to put that patient in a waiting room with other people who might not be infected, says Gehrke, who is also a professor of microbiology and immunology at Harvard Medical School. That initial triage can be very important from a public health standpoint, and there could be a follow-up test later with PCR or something to confirm. The researchers hope to obtain Food and Drug Administration approval to begin using the device in areas where the Ebola outbreak is still ongoing. In order to do that, they are now testing the device in the lab with engineered viral proteins, as well as serum samples from infected animals.

    This type of device could also be customized to detect other viral hemorrhagic fevers or other infectious diseases, by linking the silver nanoparticles to different antibodies.

    The research was funded by the National Institute of Allergy and Infectious Disease.

    Quick test for Ebola

  • Recent Research News

    The search for a new antibiotic

    -Story Source: The National UAE

    Mitya Underwood

    Image Credit: The iChip device, developed by scientists at Northeastern University, Boston, allows bacteria to grow in its natural environment but to be isolated and studied at the same time. Slava Epstein / Northeastern University

    SScientists have made a breakthrough in their search for a new generation of antibiotics. By studying bacteria in the soil they believe they can develop medicines that will combat the menace of drug-resistant superbugs. The solution to what the World Health Organisation describes as a profound threat to human health could lie at the bottom of your garden or in a muddy field. Buried in the dirt are thousands of compounds that could bring an end to superbugs the illnesses that no longer respond to most antibiotics. Scientists at Northeastern University in Boston, US, announced a breakthrough last month, in that they have worked out a way of cultivating bacteria that until now has failed to grow in laboratory conditions, but which could be the source of a new generation of antibiotics. Naturally occuring micro-organisms and bacteria are the main source of antibiotics used today. In the heyday of antibiotic discovery between the 1930s and 1970s, scientists could only study about one per cent of the bacteria found in soil samples because the other 99 per cent would not grow outside their natural environment. This made the discovery of antibiotics a difficult, costly and lengthy exercise.

    Overmining of this limited resource by the 1960s brought an end to the initial era of antibiotic discovery, said Northeastern Universitys Kim Lewis, Slava Epstein and others, in their research paper published in Nature magazine. No new major forms of antibiotics have been developed in the past 30 years, and resistance to certain types has been growing. The latest breakthrough in growing bacteria in the lab could unlock a huge source of as-yet untapped antibiotics. The success lies in the development of a technology called iChip, which allows bacteria to grow in soil, their natural environment, and to be isolated and studied at the same time. IChip involves diluting a sample mixture of soil so that one bacterial cell is placed between two laboratory slides and put back in the soil.

    The scientists estimate that almost half of samples will grow using this method, as compared to just 1 per cent of cells from soil that would grow in a lab. In this experiment the team, consisting of Lewis, Epstein and colleagues from the University of Bonn in Germany, Selcia in the UK and Novo Biotic Pharmaceuticals in Cambridge, Massachusetts, then

    Screened 10,000 samples grown in iChips for antimicrobial activity on Staphylococcus aureus, the resistant form of which is known as MRSA.A newly discovered compound that the team named teixobactin has already shown promise against resistant forms of bacteria such as S.aureus and Mycobacterium tuberculosis. It was also exceptionally active against non-resistant Clostridium difficile, which causes infectious diarrhoea, and Bacillus anthracis, the anthrax bacteria.

    Dr Anjam Khan, principal investigator and director of the Microbiology Containment Level 3 Research Suites at Newcastle University, UK, said: The good thing about this announcement isnt so much the antibiotic but the approach the investigators used to try to cultivate bacterial dark matter.Scientists have always tried to mimic environmental conditions in an artificial laboratory growth medium rather than going back to the soil.The strategy these scientists have used is very elegant. They devised a new experimental tool where they could look at individual bacteria growing in their natural soil environment.Most antibiotics we get are from soil-dwelling bacteria, yet we are missing 99 per cent of this diverse and rich population of bacteria which we simply cannot cultivate in artificial laboratory media.The technology is very simple. Thats one of the elegant and powerful things about this approach.

    He said scientists had been trying for many years to grow bacteria in a laboratory, and it had been a guessing game as to what nutrients were needed to ensure growth.

    MICROBIOZ INDIA MARCH 2015 www.microbiozindia.com 14

  • Recent Research News

    Antibiotics give rise to new communities

    Researchers have identified unexpected activity of antibiotics, potentially reshaping our understanding of why antibiotics evolved and pointing to broad health implications. Credit: Oleksii Nykonchuk / Fotolia

    News Source: Science Daily

    MMost people have taken an antibiotic to treat a bacterial infection. Now researchers from the University of North Carolina at Chapel Hill and the University of San Diego, La Jolla, reveal that the way we often think about antibiotics -- as straightforward killing machines -- needs to be revised.

    The work, led by Elizabeth Shank, an assistant professor of biology in the UNC-Chapel Hill College of Arts and Sciences as well as microbiology and immunology in the UNC-Chapel Hill School of Medicine, and Rachel Bleich, a graduate student in the UNC-Chapel Hill Eshelman School of Pharmacy, not only adds a new dimension to how we treat infections, but also might change our understanding of why bacteria produce antibiotics in the first place. "For a long time we've thought that bacteria make antibiotics for the same reasons that we love them -- because they kill other bacteria," said Shank, whose work appears in the February 23 Early Edition of the Proceedings of the National Academy of Sciences. "However, we've also known that antibiotics can sometimes have pesky side-effects, like stimulating biofilm formation."Shank and her team now show that this side-effect -- the production of biofilm -- is not a side-effect after all, suggesting that bacteria may have evolved to produce antibiotics in order to produce biofilm and not only for their killing abilities.Biofilms are communities of bacteria that form on surfaces, a phenomenon dentists usually refer to as plaque. Biofilm are everywhere. In many cases, biofilm can be beneficial, such as when they protect plant roots from pathogens. But they can also harm, for instance when they form on medical catheters or feeding tubes in patients, causing disease."It was never that surprising that many bacteria form biofilm in response to antibiotics: it helps them survive an attack. But it's always been thought that this was a general stress response, a kind of non-specific side-effect of antibiotics. Our findings indicate that this isn't true. We've discovered an antibiotic that very specifically activates biofilm formation, and does so in a way that has nothing to do with its ability to kill."Shank and her team previously reported that the soil bacterium Bacillus cereus could stimulate the bacterium Bacillus subtilis to form a biofilm in response to an unknown secreted signal. B. subtilis is found in soil and the gastrointestinal tract of humans. Using imaging mass spectrometry, they subsequently identified the signaling compound that induced biofilm production as thiocillin, a member of a class of antibiotics called thiazolyl peptide antibiotics, which are produced by a range of bacteria. At that point, Shank and her colleagues knew thiocillin had two very specific and different functions, but they didn't know why -- and wanted to know how it worked. That's when they modified thiocillin's structure in a way that eliminated thiocillin's antibiotic activity, but did not halt biofilm production. "That suggests that antibiotics can independently and simultaneously induce potentially dangerous biofilm formation in other bacteria and that these activities may be acting through specific signaling pathways," said Shank. "It has generated further discussion about the evolution of antibiotic activity, and the fact that some antibiotics being used therapeutically may induce biofilm formation in a strong and specific way, which has broad implications for human health."

    MICROBIOZ INDIA MARCH 2015 www.microbiozindia.com 15

  • Recent Research News This is a reconstructed visual of dengue virus serotype 3 bound with antigen-binding fragments of super-potent antibody 5J7. Credit: Guntur Fibriansah

    News Source: Science Daily

    AA new Duke-NUS-led study has identified a super-potent antibody which requires a minute amount to neutralize the dengue virus. The study, published online on 20 February 2015, in the journal Nature Communications, showed how a newly identified antibody 5J7, is highly effective in killing dengue virus whereby only 10-9 g of antibody is needed to stop the infection of dengue serotype 3 virus (DENV-3). This new finding gives hope for the development of effective dengue treatments.

    Over the last 50 years, the incidence of dengue virus has increased by 30 times worldwide. The virus causes fever, rashes and joint pain and in severe cases, bleeding and shock. It is estimated to be endemic in 100 countries and is a huge burden on healthcare systems. However, till now, there is no licensed dengue vaccine or therapeutic agent due to the presence of four circulating virus serotypes (DENV1-4) complicating their development.

    Senior author Associate Professor Shee Mei Lok from Duke-NUS Graduate Medical School Singapore (Duke-NUS) focuses her research on understanding the pathology and structure of the dengue virus to develop effective therapeutics. Her lab has already discovered antibodies that are effective against DENV-1. Her strategy to develop a safe therapeutic is to combine four antibodies that each bind and potently inhibit infection of each of the dengue virus serotypes.

    In this recent study, researchers isolated 5J7 from 200 different candidate antibody molecules by studying blood samples from a dengue infected patient. By examining the virus-antibody complex structure at very high magnification, they showed that each arm of the antibody is surprisingly effective in grabbing three surface proteins on the surface of the virus at the same time. In addition, the sites on the virus where the antibody was bound were critical for the virus to invade cells.

    "This kind of binding with the virus has never been observed and it explains why the antibody itself is so highly potent." said A/Prof Lok, who is from the Emerging Infectious Diseases Programme at Duke-NUS. "The movement of virus surface proteins is highly essential for invading cells -- you can think of antibody 5J7 locking the virus surface proteins, thus strapping the virus."

    While antibody 5J7 has been found to be effective against DENV-3, the remaining two serotypes of dengue virus (DENV-2 and DENV-4) have to be considered. When a patient is infected by one serotype -- this stimulates the production of a variety of antibodies that kills that serotype and that patient will have life-time immunity towards that particular serotype. However, in this process, the patient will also produce antibodies that will bind the other three if they are infected by them. This may enhance their secondary infection and cause the development of a more severe form of the disease.

    "We need to test the efficacy in mouse models first and then move to clinical trials," said A/Prof Lok about the next step after this promising finding. "We are optimistic that we will make a treatment breakthrough within these few years but antibodies against all the other serotypes have to be identified first."

    MICROBIOZ INDIA MARCH 2015 www.microbiozindia.com 17

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  • Recent Research News Researchers introduce the idea of using sewage to study human microbiome Story Source: Marine Biological

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    AA new study demonstrates that sewage is an effective means to sample the fecal bacteria from millions of people. Researchers say the information gleaned from the work provides a unique opportunity to monitor, through gut microbes, the public health of a large population without compromising the privacy of individuals.

    Humans harbor tremendous amounts of bacteria in their gastrointestinal tract and gut bacteria serve important functions in healthy humans. Studies of the human microbiome, the collection of trillions of microbes living in and on the human body, have gained traction during the last decade. There is a great interest in identifying a "healthy microbiome" by identifying one or more bacterial community types that may be associated with healthy individuals, however financial considerations and privacy concerns limit the number of individuals who can be screened.

    In a new study published in the January/February 2015 issue of the journal mBio, researchers from the Marine Biological Laboratory (MBL) and the University of Wisconsin-Milwaukee (UWM) School of Freshwater Sciences introduce the idea of using sewage as a population level pool that carries a signal for the microbiomes of humans.

    Using oligotyping, a novel approach developed at the MBL, scientists compared the gut bacterial community profiles of 137 healthy adults provided by The Human Microbiome Project to the bacterial community profiles of more than 200 sewage influent samples collected from 71 U.S. cities. In the paper led by UWM's Ryan Newton, researchers found that geographically distributed populations share a small core set of bacteria whose members represent various common community states within U.S. adults. The study uses the percent of obese individuals in a given city as a measure of lifestyle differences across cities, and demonstrates that the bacterial community structure is a good predictor, with 81 to 89 percent accuracy, of a city's estimated level of obesity. Lifestyle differences can reproducibly alter the human gut microbiome, and microbial community composition is a known indicator of obesity.

    "This method is similar to trying to create a map of a geographical region," explains A. Murat Eren, an Assistant Research Scientist at the MBL, and one of the authors of the study. "The way we have been working with microbiomes of individuals has been similar to driving around and mapping the streets and structures of a city in a detailed manner. This approach takes our efforts to a much larger scale. In this sense it is similar to taking one big aerial picture of a city, trading off intricate details of a small number of well-described streets for broader insights and larger patterns." The researchers say the use of oligotyping, which provides greater sensitivity, allowed them to better explain the distribution of very closely related bacterial organisms to compare microbiomes among 71 human populations."The sewage samples of 71 cities do not tell us anything specific about 'individuals' who live in those cities" says Eren. "However, only using sewage samples, we were able to differentiate these cities based on their estimated level of obesity. This approach can be beneficial to answer various public health questions while not compromising the privacy of individuals. For instance, microbial observatories plugged into sewage systems can keep us informed about the general health of large populations without being intrusive." "This work fits into our long-term goal of developing better water pollution and public health assessments," says UWM professor and study co-author Sandra McLellan. "It's a great example of how new sequencing technologies and novel computational approaches can allow us to glean new information from complex environments."

  • Recent Research News

    Quick test for quality beer, milk

    Story Source: Science daily

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    TTo guarantee a high quality of their beer, breweries monitor the production process very closely. With a new polymer powder, this monitoring will be able to be faster and simpler in the future. Manufacturers can also test drinks such as milk, juice, cola and red wine with the quick check. It tastes full-bodied and spicy, is tasty and is welcome refreshment, especially in the hot summer months -- Beer is very popular throughout the world. For brewers, a consistently high quality of the drink is essential. To ensure this, the companies try to keep the product free from harmful microorganisms. This is because pathogens that enter into the beer during the brewing process can spoil the pleasure of the drink. They not only provide strong variations in taste and smell; the beer can also become cloudy, sour and unwholesome. Therefore, ongoing quality controls accompany the production process. However, conventional microbiological methods require five to seven days to detect beverage-spoiling organisms, such as bacteria and yeasts. It is often too late at that point to take corrective action. In collaboration with the company GEN-IAL from Troisdorf, researchers at the Fraunhofer Institute for Applied Polymer Research IAP in Potsdam have developed a polymer powder that significantly simplifies these tests and shortens the time that they require. The company supplies breweries with analysis tools for quality control. From the test to the reliable result takes two to three days. The reason: Until recently, beer has been filtered in special equipment. In this process, the bacteria remain on a membrane and are then elaborately cultivated in a special culture medium before they can be examined microscopically. The new polymer powder from the IAP replaces this process: The powder is added to the liquid sample. The powder's functionalized surface binds the bacteria efficiently. The pathogens adhere to the 100 to 200 micron powder particles. These can be easily removed along with the microbes in a specially developed system and analyzed directly using various microbiological methods. The time-consuming enrichment in a nutrient medium is no longer necessary.

    Quality control of large quantities of beverages possible: With the new method, food experts can investigate beer and other beverages for infection by pathogens, which was hardly or not at all possible with the traditional membrane filtration method. "Membrane filtration is not suitable for the quality control of beverages such as fruit juices, milk, cola and red wine. They contain so much solid or suspended matter that the filter clogs quickly," explains Dr. Andreas Hollnder, scientist at the IAP. Breweries have also only been able to examine small sample volumes of up to one liter via membrane filtration. With the polymer powder, tests with 30 liters or more are possible. "Wherever a small amount of microbes has to be extracted from a large amount of liquid, the new technique can be useful," adds Hollnder. "Through the use of the powder, food safety is increased, since it is more likely to find trace contaminants in large volumes of the beverages," says Dr. Jutta Schnling, managing director of Gen-IAL. From the test to the reliable result takes two to three days. The reason: Until recently, beer has been filtered in special equipment. In this process, the bacteria remain on a membrane and are then elaborately cultivated in a special culture medium before they can be examined microscopically. The new polymer powder from the IAP replaces this process: The powder is added to the liquid sample. The powder's functionalized surface binds the bacteria efficiently. The pathogens adhere to the 100 to 200 micron powder particles. These can be easily removed along with the microbes in a specially developed system and analyzed directly using various microbiological methods. The time-consuming enrichment in a nutrient medium is no longer necessary.

    Quality control of large quantities of beverages possible :With the new method, food experts can investigate beer and other beverages for infection by pathogens, which was hardly or not at all possible with the traditional membrane filtration method. "Membrane filtration is not suitable for the quality control of beverages such as fruit juices, milk, cola and red wine. They contain so much solid or suspended matter that the filter clogs quickly," explains Dr. Andreas Hollnder, scientist at the IAP. Breweries have also only been able to examine small sample volumes of up to one liter via membrane filtration. With the polymer powder, tests with 30 liters or more are possible. "Wherever a small amount of microbes has to be extracted from a large amount of liquid, the new technique can be useful," adds Hollnder. "Through the use of the powder, food safety is increased, since it is more likely to find trace contaminants in large volumes of the beverages," says Dr. Jutta Schnling, managing director of Gen-IAL.

    MICROBIOZ INDIA MARCH 2015 www.microbiozindia.com 20

  • Recent Research News

    Asian herb holds promise as treatment for Ebola virus disease

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    NNew research that focuses on the mechanism by which Ebola virus infects a cell and the discovery of a promising drug therapy candidate is being published Feb. 27, 2015, in the journal Science. Dr. Robert Davey, scientist and Ewing Halsell Scholar in the Department of Immunology and Virology at Texas Biomedical Research Institute announced today that a small molecule called Tetrandrine derived from an Asian herb has shown to be a potent small molecule inhibiting infection of human white blood cells in vitro or petri dish experiments and prevented Ebola virus disease in mice. The latest outbreak of Ebola virus disease has caused the death of more than 9,400 people worldwide and created an international crisis that has shown few signs of stopping, continuing to infect thousands in West Africa. Ebola virus causes hemorrhagic fever in humans and currently has no approved therapy or vaccine. Scientists at Texas Biomed have been working in the Institute's Biosafety Level 4 containment laboratory for more than 10 years to find a vaccine, therapies and detection methods for the virus.

    Davey and his team have been working for more than five years on identifying and finding therapy targets for Ebola virus disease. Davey's research has focused on stopping the virus before it has a chance to enter or interact with cellular factors, as that is a critical first step to combating infection. Ebola virus begins its entry into a cell by first binding to several types of cell surface proteins. Then the virus is taken into the cell and follows an endosomal route, or membrane-bound route that transports the virus to various cell compartments.

    From previous studies, Davey said that during this endosomal process, he knew that calcium signaling in cells, which allow cells to transmit electrical charges to one another, controls many of the processes in the cell and was important for Ebola virus infection."We were not able, however, to pinpoint the mechanisms involved in this process," Davey explained. "With this research, we discovered that two pore channels (TPCs) are the key calcium sensor involved in Ebola virus infection. These TPCs essentially needed to be turned on in order for the virus to function properly."Two pore channels are unusual calcium channels found in endosomes that control the way endosomes move through cells and the environment of the cells. Davey compared TPCs to traffic cops and air conditioners, helping direct the endosomes and any virus it might be carrying through the cell and making the endosomes and its passengers more comfortable along the way. Davey and his team were able to show the critical role of two pore channels in Ebola virus infection, which has not previously been shown in any other virus. In addition to identifying this critical mechanism to infection, Davey's team also showed that drugs targeting this interaction show some efficacy as potential treatments against Ebola virus disease. In the study, Davey's team determined that existing drugs currently used to treat high blood pressure have an ability to turn this key calcium sensor on and off. Working with a group in Munich, Germany and Southwest Research Institute, the team tested several small molecules to see which was most effective at turning the sensors off thus prohibiting Ebola virus from moving any further through the cell.

    The team found Tetrandrine protected mice from disease without obvious side effects and was the best candidate for further animal testing, because it was the most potent compound tested, gave little evidence of cytotoxicity and required a smaller dose to be effective and tolerated.

    "When we tested in mice, the drugs stopped virus replication and saved most of them from disease," Davey said. Essentially, this drug shows an ability to stop the virus before it has a chance interact with cellular factors, thus stopping the virus from continuing its infection process."We are very excited about the progress made in this study and the momentum it provides as scientists across the world vigorously search for effective vaccines and treatments against Ebola virus," Davey said. "We are cautiously optimistic. The next step in the process is to test both safety and effectiveness of the interaction of the drug with Ebola virus in non-human primates."

    MICROBIOZ INDIA MARCH 2015 www.microbiozindia.com 21

  • National Conference on

    RECENT TRENDS IN APPLIED MICROBIOLOGY, HUMAN HEALTH, & ENVIRONMENT

    March 27-28, 2015 (Sponsored by University Grants Commission, New Delhi)

    Organized by DEPARMENT OF MICROBIOLOGY,

    BUNDELKHAND UNIVERSITY, JHANSI The uniqueness of Microorganisms and their often unpredictable nature has made them likely candidates for solving difficult problems in the life sciences and other field of environment,Agriculture,Industries,Medical and other fields which directly or indirectly influence humankind ,Neverthless,while microbial biotechnologies have been applied to various problems of above mentioned fields with considerable success in recent years. they have not been widely accepted by the scientific community because it is often difficult to consistently reproduce. This seminar provides an opportunity to discuss recent developments in the understanding of how microorganisms can interact in Agriculture and natural ecosystems, Microbial applications in industrial and Medical fields will also discussed.

    Conference Theme

    Programme will include invited guest lectures and presentation in following area:

    Microorganisms in sustainable Environment Microorganism in Sustainable Agriculture Microorganism in Industries Microorganism in Medical sciences Other application of Microbes Instructions for abstract preparation

    Conference is open to all involved in area of conference theme, interested participants are invited to contribute their papers for oral/or poster presentation. Abstracts should be prepared (about 250 words) using MS-Words and sent to the [email protected] as file attachment. The name of presenting author should be underlined.

    Instructions for full paper presentation The full paper should include a title with the full name(s) of author(s), Affiliation(s) where the work was carried, an Abstract followed by sections including an introduction, the main body of text,conclusions,acknowledgements and references.

    Schedule of events March 15, 2015 Deadline of receipt of Abstract march 20, 2015 Deadline for submission of full papers March 27-28, 2015 Conference

    Registration

    Chief Patron Prof.A.C. Pandey,Honble Vice Chancellor,Bundelkhand University,Jhansi Patron Prof. Pankaj Atri,Pro Vice Chancellor,Bundelkhand University,Jhansi Chairperson Prof.S.P.Singh,Dean Science,B.U Organizing Secretary Dr.Rishi Kumar Saxena,Head,Dept of Microbiology,B.U.Jhansi Co-Organizing Secretary Dr.Devendra Mani Tripathi and Dr.Sanjay Kumar Joint Secretary Dr.Sangeetalal Co-Joint Secretaries Mr.Pankaj Sagar & Dr.Ranjana

    Organizing Committee

    Participants are expected to register on or before March, 15,2015,by sending the requisite fee in form of DD in favour of the organizing Secretary, National Conference on Recent Trends in AppliedMicrobiology,Human Health & Environment, Payable at SBI,BU Campus, Jhansi along with requisite for accommodation, if required. Invited Speakers may register on arrival.

    Participants On time Late

    Faculty Members Rs. 700/- Rs.1000/-

    On time Late

    Students/research scholar Rs.500/- Rs.700/-

    Address for correspondence

    Dr.Rishi Kumar Saxena

    Head & Organizing Secretary of National Conference

    Department of Microbiology,

    Bundelkhand University, Jhansi-284128

    E-mail:[email protected]

    Mob:+91-9889106832

    Fax: 0610-2321667

  • Recent Research News

    Living in genetic comfort zone: How to avoid influence of genetic variation

    Story Source: Science daily

    TThe phenotype of organisms is shaped by the interaction between environmental factors and their genetic constitution. A recent study by a team of population geneticists at the Vetmeduni Vienna shows that fruit flies live in a sort of genetic comfort zone at a specific temperature. The scientists found that, despite their underlying genetic differences, two separate strains of flies had a very similar gene expression pattern at 18C. This effect of 'canalization', which has also been described in humans, allows organisms to continue to grow and develop stable even in the face of genetic and environmental stress. The results were published in the journal PLOS Genetics. The information encoded in the DNA of an organism is not sufficient to determine the expression pattern of genes. This fact has been known even before the discovery of epigenetics, which refers to external modifications to the DNA that turn genes "on" or "off." These modifications do not change the DNA sequence, but instead, they affect how genes are expressed. Another, less known mechanism called canalization keeps organisms robust despite genetic mutations and environmental stressors. If an organism experiences environmental or genetic perturbations during its development, such as extreme living conditions or genetic mutations, canalization acts as a way of buffering these disturbances. The organism remains stable and can continue to develop without recognizable changes.

    A comfort zone in the fly genome

    Christian Schltterer at the Institute of Population Genetics and his colleagues studied the mechanism of canalization in fruit flies. The researchers subjected two genetically distinct strains of fruit flies, Oregon and Samarkand, to different temperatures (13C, 18C, 23C and 29C). Subsequently, they analysed the variation in gene expression in response to the different temperatures. The results revealed a homogenous pattern of gene expression among the two strains at 18C. No matter whether the flies were from the Oregon or to the Samarkand strain, their gene expression was almost indistinguishable.

    "The flies' genetic comfort zone appears to be located at 18C. "As soon as the flies leave the comfort zone, move to either higher or lower temperatures, the gene expression of the two strains varies dramatically" Schltterer explains.

    Buffering the genotype

    The effect of canalization was first described in 1942, when researchers pointed out that organisms remain stable in their external appearance despite different environmental circumstances or genetic mutations. This sort of developmental buffering helps to stabilize organismal growth.

    "If an organism develops along the canalization pathway, or along the comfort zone, mutations can accumulate without being expressed. Once an organism leaves the canalized range, those hidden genetic variations can be expressed and become visible. The phenomenon is called Decanalization," Schltterer explains.

    Decanalization as the origin of complex genetic disease

    A publication by U.S. researcher Greg Gibson in the journal Nature (Paper-Link) proposes that diseases such as diabetes, asthma, depression and cardiovascular disease are the consequence of genetic Decanalization. He describes how migration, diet, smoking, air pollution and psychological stress can lead to stress-mediated Decanalization and therefore cause certain complex genetic diseases in humans.

    "Genetic information alone does not determine whether we stay healthy or not. It is the complex interaction of environmental conditions and genetic variation that needs to be considered," says Schltterer.

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  • Recent Research News

    Tracking parasites with satellites

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    SScientists are teaming up to use satellite data to target deadly parasites to help predict patterns of parasitic diseases such as malaria, worms and hydatids. Project leader Professor Archie Clements, from The Australian National University, said the research could help authorities in developing countries fight parasitic diseases.

    "Some diseases are highly sensitive to their environment, especially parasitic diseases. With remote sensing you can identify places where disease flourishes," said Professor Clements, Director of the ANU Research School of Population Health."This information is useful for decision makers to help them ensure scarce resources are targeted to where they are most needed."Parasitic diseases affect hundreds of millions of people every year, many of them in the least developed parts of the world. The team uses satellite data such as temperature, rainfall, vegetation and land usage, and combines it with health data in a geographical information system (GIS).The approach combines the skills of many scientists, such as entomologists, epidemiologists, software developers, social scientists and health policy specialists."The result is maps that are accessible to countries with limited capacity for managing disease data, tailored to their local needs."The team has trialed systems for malaria in Bhutan, Vanuatu and the Solomon Islands and is now seeking support to scale up to larger countries. Additionally, spatial predictions for other diseases such as worms and hydatids are being developed for China, the Philippines and other countries in the Asia-Pacific region."By taking this research the next step, we have the opportunity to have a meaningful impact on the real world, and save a lot of lives," Professor Clements said. Professor Clements is laying out a plan for the future of these systems at a symposium at the American Association for the Advancement of Science Conference, in San Jose, California this weekend.

    Story Source: Science daily

  • Recent Research News

    Malaria transmission linked to mosquitoes' sexual biology

    Story Source: Science daily

    SSexual biology may be the key to uncovering why Anopheles mosquitoes are unique in their ability to transmit malaria to humans, according to researchers at Harvard T. H. Chan School of Public Health and University of Perugia, Italy. Through analysis of 16 Anopheles genomes, they found that these mosquitoes' reproductive traits evolved along with their capacity to transmit the Plasmodium parasite that causes malaria. These findings may provide a new target for malaria control, particularly in regions hardest hit by the disease. "Our study is the first to reveal the evolutionary dynamics between the sexes that are likely responsible for shaping the ability of Anopheles mosquitoes to transmit malaria to humans," said senior author Flaminia Catteruccia, associate professor of immunology and infectious diseases at Harvard Chan School and University of Perugia.

    The study was published online February 26, 2015 in Science.

    Anopheles mosquitoes are the only mosquitoes capable of transmitting human malaria; however, the species within this genus vary widely in their ability to do so, for reasons that remain unknown. The researchers analyzed nine globally dispersed Anopheles species, enabling reconstruction of the evolutionary history of their reproductive traits and capacity to transmit malaria. They found that two key male reproductive traits in Anopheles are acquired and evolved together over time: transferring ejaculate as a gelatinous rod-shaped structure called the mating plug, and the ability to synthesize a steroid hormone contained in that plug called 20-hydroxyecdysone (20E). The researchers also demonstrated that the evolution of these male traits drove reciprocal adaptations in females strongly linked to the mosquitos' capacity to transmit malaria. This study adds to previous findings from this research group showing that sexual transfer of 20E induces a series of dramatic changes in the female, fundamentally altering her physiology and behavior. These changes affect a female's reproductive output, longevity and immune response to Plasmodium parasites, all key factors in malaria transmission. All four species of Anopheles mosquitoes that transfer large levels of 20E are major malaria vectors originating from Africa and India, the regions of highest malaria burden.

    The findings may also be applicable to Dengue and West Nile virus, which are transmitted by the Aedes and Culex mosquitoes, respectively. In these species some aspects of reproductive biology are similar to Anopheles.

    By identifying factors critical for increasing the ability of mosquitoes to transmit malaria, compounds developed to specifically target those factors could be incorporated into existing mosquito control technologies, boosting their overall effectiveness.

    MICROBIOZ INDIA MARCH 2015 www.microbiozindia.com 25

  • Recent Research News

    Pollution is driving force behind growth of nuisance algal scums linked to disease

    Story Source: Science daily

    PPotentially toxic microbes which pose a threat to our drinking water have undergone a dramatic population explosion over the last 200 years as a result of pollution, research involving experts from The University of Nottingham has found. The study, published in the journal Ecology Letters, looked at more than 100 lakes in lowland and alpine areas of North America and Europe and found that populations of Cyanobacteria -- also known as blue-green algae -- have significantly increased since the 1800s.The research, conducted in collaboration with academics at McGill University in Canada and other collaborators, is the first study to show that a rise in the algae's available nutrient sources nitrogen and phosphorus -- commonly resulting from industrial fertilisers and sewage discharge -- is the biggest potential culprit responsible for the increase in such a large number of lakes, across such a large geographical area. The study also found that climate change can exacerbate this problem, with water management challenges likely to increase in a future warmer world. Colonies of blue-green algae pose a serious threat to drinking water sources worldwide because many types contain toxins which can cause damage to the liver and nervous system and have been linked with neurodegenerative diseases such as Alzheimer's, Parkinson's, ALS and Lou Gehrig's disease. Most municipal water treatment plants do not regularly look for Cyanobacteria toxins in the water supply. However, municipalities with a known history of blooms typical monitor their surface water supplies for Cyanobacteria. When detected, the cells can be removed by adding chemicals that bind them together, so they can be separated out. Although this removes the cells, the cells may already have broken down releasing toxins into the water.

    In addition, environmental costs associated with this alga were estimated to exceed $100 million per year in both the UK and Australia. PhD researchers Heather Moorhouse and Mark Stevenson, based in the School of Geography at the University, and Dr Suzanne McGowan, Head of the School of Geography at University of Nottingham Malaysia Campus, took sample cores of sediment from lakes located in the major lake districts of the British Isles including the English Lake District, the West Midland Meres, Scottish lochs and upland lakes in Northern Ireland and analysed them for pigments left behind by blue-green algae. These pigments remain stable over thousands of years and act as biomarkers revealing the past levels of algae found in the water during the course of decades. The analysis showed that during the last 200 years, more than half of the lakes (58 per cent) had seen significant increases in concentrations of blue-green algae pigments, whereas only three per cent showed a significant decrease in the presence of the microorganism. Lowland lakes in agricultural catchments typical of those found in the UK were found to be especially susceptible to Cyanobacteria increases. The study also found that since 1945 the incidence of blue-green algae has increased more rapidly than the growth of other types of water-borne algae.

    More significant increases were observed in the more temperate lowlands (61 per cent in North America and 70 Per cent in Europe) which were closer to areas of agricultural activity than in alpine areas (36 per cent).Analysis of the trends in blue-green algae spanning 10 countries across three continents showed that growth was mainly regulated by a change in the amount of nutrients -- phosphorus and nitrogen -- found within the lakes.

    The research underlines the importance of further study into how to more carefully control the inputs of nitrogen and phosphorus-rich pollutants and ways of monitoring the resulting toxins in our drinking water. It also demonstrates that effective catchment nutrient control initiated by legislation such as the EU Water Framework Directive is important to the safety of our water supplies in the future.

    Image Credit: New York State, Dept of Environment Conservation

    MICROBIOZ INDIA MARCH 2015 www.microbiozindia.com 26

  • Interview An Interview with Prof. Aussielita L. Lit, Philippines, Under Microbioz India Scientist speak

    Microbioz Team: Why you opt MICROBIOLOGY as a career? Prof. Aussielita: Honestly, I was able to pursue studies in the field of Microbiology because first, Its my personal decision to go on this field. I had already experienced as early as during my high school days preliminary trainings and basic knowledge on this field. So, I decided to continue learning its concepts and applications until I finished my Baachelors Degree in one of the most prestigious universities here in Philippines and precede Masters Studies. I also look it as very significant area of study in that it deals with microbes and microorganism-causing diseases which we can reflect on the present epidemia of some diseases like HIV Virus, Mers-Cov and the Ebola virus. For me, its really a perfect course which makes me believe that I am born to be one. Microbioz Team: Tell us a little more about your professional experiences; particularly those not mention your resume/application? Prof. Aussielita: Aside from the obvious information about my education attainment, pursuing with Microbiology was really adventurous and intellectual for me and it will always be. I have my specialization on Food Microbiology and Industrial Microbiology. Its adventurous in that I visited some profound and beautiful places which I only dreamed of before, this was when I attended some workshops, seminars and large group scientific conferences outside country. It brought me also into another dimension of learning in that as the longer I studied it the more I become efficient scientist and microbiologist. My profession really inspired me a lot. It realized me how important is to deal with some aspects of society especially health and safety of the people. Also, I availed international studies because of the advantage brought by studying Microbiology. Microbioz Team: What is your favorite part of your current job and why is it your favorite part? Prof. Aussielita: The most exciting part of my current job as a Laboratory Manager is that I enjoyed providing guidance to entrees and my research students and advisees on their research works. It makes me happy and honored when someone asks my help and I could provide them for their betterment. Microbioz Team: How would your background and experiences strengthen this academic and research platform? Prof. Aussielita: The laboratory I managed was founded to strengthen infrastructure and support services in the agro-industrial sectors and to develop the quality of life especially here in the locality. Moreover, I am glad that this research platform holds complec and systematized functions because I have my colleagues, one who is a chemical engineer and some are administrative staffs and my research assistants. Microbioz Team: What is one or two of your proudest professional accomplishments? Prof. Aussielita: Throughout my profession experience, I must say that I have completed more than halfway of my dreams. My proudest accomplishments were first, when I declared as a Registered Microbiologist of the Philippine Academy for Microbiology dated 1994. . It all began on this step. Another was when I have Utilized the significant concepts of Microbiology, applied those concepts and successfully find noble employment. Evidence to these were my experience as specialist microbiologist in one of the big Food Industries here in Philippines ( DOLE Philippines) , became a technical and quality asessor and made my own employment by establishing Research Laboratory. Microbioz Team: Mention few of your words in favour of Microbioz India. Prof. Aussielita: I find it as a very good source or reference material for some science people and researchers. It also amazed me that it also launches interviews for some scientists and biologists. You could really provide good information to our people. More success to Microbioz India Magazine.

    Ma. Aussielita L. Lit (Uchie) is a registered microbiologist of the Philippine Academy for Microbiology in 1994 and became a specialist microbiologist in 2002. She is a recognized signatory for chemical and microbiological testing by the Philippine Accreditation Office (PAO) of the Department of Trade and Industry (DTI) since 1995. She had 17 years of private practice as microbiologist in one of the big Food Industries in the country. Recently established her own Research Laboratory.

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    RRooddeell EEssttaaddiilllloo AAlloo,, RReesseeaarrcchheerr PPhhiilliippppiinneess ((MMiiccrroobbiioozz RReepprreesseennttaattiivvee))

    MICROBIOZ INDIA MARCH 2015 www.microbiozindia.com 27

  • Find your Scholarships updates here!!!

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  • Holland Scholarship for International Students in Netherlands, 2015

    About Scholarship Applications are invited for Holland Scholarship for international students from outside the European Economic Area (EEA).

    Scholarship is awarded for pursuing bachelors or masters programmes at one of the participating Dutch higher education institutions. The scholarship amounts to 5,000. Student will receive this in the first year of their studies. The application deadline is 31 March, 2015.

    Eligibility Your nationality is non-EEA. You are applying for a full-time bachelors or masters at one of the participating Dutch higher education institutions. You meet the specific requirements of the institution of your choice. You have never before studied at an education institution in the Netherlands.

    How to Apply You can apply for the Holland Scholarship at the Dutch institution of your choice. The institution will select those who will be

    granted the scholarship. You can find an overview of participating Dutch research universities and universities of applied sciences, as well as selected fields of study.

    Deadline The application deadline is 31 March, 2015.

    For Details http://www.studyinholland.nl/scholarships/holland-scholarship

    Open scholarships

    MICROBIOZ INDIA MARCH 2015 www.microbiozindia.com 29

  • 2015 CiM-IMPRS PhD Fellowships in Life and Natural Sciences, Germany

    About Scholarship The International Max Planck Research School Molecular Biomedicine (IMPRS-MBM) and the Cells in Motion Excellence Cluster

    (CiM) offer 16 PhD Fellowships in Life and Natural Sciences. Applications are invited for highly qualified and motivated students of any nationality from biological sciences, chemistry, mathematics, computer sciences and physics.

    Eligibility Applicants must hold a Masters degree, or equivalent (e.g. German Diploma), in life or natural sciences (e.g. biology,

    biochemistry, biotechnology, physics, chemistry, pharmacy or related fields), in mathematics or computer sciences. Applicants with a state examination in pharmacy can be admitted to work on pharmaceutical/chemical projects. The date of award of the Masters (diploma, state examination) degree must not be more than 4 years ago (i.e. 4 years before the above-mentioned application deadline).

    Applications are also welcome if the required degree has not yet been awarded by the time of application. However, it has to be awarded before fellowships commence in October.

    How to Apply Applications can only be submitted via online

    Applicants should first go to our webpage to see ALL details about the application process before they apply online.

    Deadline Candidates will be notified on 29 June 2015.

    For Details www.cim-imprs.de

    HEC Fully Funded PhD Scholarships for University of Cambridge in UK, 2015

    Open scholarships

    MICROBIOZ INDIA MARCH 2015 www.microbiozindia.com 30

  • About Scholarship The HEC and Cambridge Commonwealth Trust (CCT) jointly fund PhD scholarships for University of Cambridge. Scholarships are

    awarded for Pakistani students to undertake direct PhD & MS/MPhil leading to PhD Program at University of Cambridge.

    Eligibility Pakistani nationals Candidates must have minimum sixteen years of education. Maximum two second divisions throughout the academic career Maximum age on Friday May 15, 2015: