This document describes a study that isolated lactic acid bacteria (LAB) from various plant and vegetable matrices. 22 LAB strains were isolated belonging to Lactobacillus, Lactococcus, and Enterococcus genera. The predominant species were Lactobacillus brevis (57%) and Lactobacillus delbrueckii subsp. bulgaricus (14%). The strains were tested for their ability to produce bacteriocin-like substances (BLS) with antimicrobial activity. A conjugation experiment was then successfully performed between Enterococcus faecium and Lactobacillus acidophilus to demonstrate horizontal gene transfer of BLS production.
Isolation, Identification of Probiotic Bacteria Present in Milkijtsrd
The aim ofThe aim of this study was to present some data on isolation, growth, and antimicrobial activity, effect of pH, heat, and sensitivity to proteolytic enzymes of lactobacillus as probiotic bacteria. A large amount of probiotic bacteria is present in milk or milk products which are mainly lactic acid bacteria (LAB). Lactic acid bacteria (LAB) including Lactobacillus spp. Isolation, identification of lactic acid bacterial (LAB) was done by Gram' staining and catalase test and further confirmation was based on morphological, cultural, physiological and different biochemical tests. A total four isolates viz. Lactobacillus fermentum, L. casei, L. acidophilus and bifid bacterium longum was identified after different biochemical analysist which were also showed reliable probiotic properties. The antibacterial activity of lactic acid bacteria (LAB) isolated from raw milk against common enteric pathogens. The antagonistic properties of these isolates against Escherichia coli, Staphylococcus aureus, were examined using agar well diffusion method. this study was to present some data on isolation, growth, and antimicrobial activity, effect of pH, heat, and sensitivity to proteolytic enzymes of lactobacillus as probiotic bacteria. A large amount of probiotic bacteria is present in milk or milk products which are mainly lactic acid bacteria (LAB). Lactic acid bacteria (LAB) including Lactobacillus spp. Isolation, identification of lactic acid bacterial (LAB) was done by Gram' staining and catalase test and further confirmation was based on morphological, cultural, physiological and different biochemical tests. A total four isolates viz. Lactobacillus fermentum, L. casei, L. acidophilus and Bifidobacterium longum was identified after different biochemical analysist which were also showed reliable probiotic properties. The antibacterial activity of lactic acid bacteria (LAB) isolated from raw milk against common enteric pathogens. The antagonistic properties of these isolates against Escherichia coli, Staphylococcus aureus, were examined using agar well diffusion method. Shobha Mehra | Vimla Mehra | Dinesh Bhauryal"Isolation, Identification of Probiotic Bacteria Present in Milk" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-2 | Issue-5 , August 2018, URL: http://www.ijtsrd.com/papers/ijtsrd16979.pdf http://www.ijtsrd.com/biological-science/biotechnology/16979/isolation-identification-of-probiotic-bacteria-present-in-milk/shobha-mehra
MPN is most commonly applied for quality testing of water, i.e., to ensure whether the water is safe or not in terms of the bacteria present in it. The presence of very few faecal coliform bacteria indicates that the water is likely free of disease-causing organisms.
video link: https://youtu.be/vOB9AQo54vU
viable but non-culturable microorganisms (VBNC).
WHAT IS A VBNC STATE?
CHARACTERISTICS OF BACTERIA IN VBNC STATE.
CONDITIONS STIMULATING VBNC STATE.
METHODS FOR DETECTION OF VBNC STATE.
Impact on public health.
Advantages of VBNC.
Disadvantages of VBNC state.
Conclusion.
Isolation, Identification of Probiotic Bacteria Present in Milkijtsrd
The aim ofThe aim of this study was to present some data on isolation, growth, and antimicrobial activity, effect of pH, heat, and sensitivity to proteolytic enzymes of lactobacillus as probiotic bacteria. A large amount of probiotic bacteria is present in milk or milk products which are mainly lactic acid bacteria (LAB). Lactic acid bacteria (LAB) including Lactobacillus spp. Isolation, identification of lactic acid bacterial (LAB) was done by Gram' staining and catalase test and further confirmation was based on morphological, cultural, physiological and different biochemical tests. A total four isolates viz. Lactobacillus fermentum, L. casei, L. acidophilus and bifid bacterium longum was identified after different biochemical analysist which were also showed reliable probiotic properties. The antibacterial activity of lactic acid bacteria (LAB) isolated from raw milk against common enteric pathogens. The antagonistic properties of these isolates against Escherichia coli, Staphylococcus aureus, were examined using agar well diffusion method. this study was to present some data on isolation, growth, and antimicrobial activity, effect of pH, heat, and sensitivity to proteolytic enzymes of lactobacillus as probiotic bacteria. A large amount of probiotic bacteria is present in milk or milk products which are mainly lactic acid bacteria (LAB). Lactic acid bacteria (LAB) including Lactobacillus spp. Isolation, identification of lactic acid bacterial (LAB) was done by Gram' staining and catalase test and further confirmation was based on morphological, cultural, physiological and different biochemical tests. A total four isolates viz. Lactobacillus fermentum, L. casei, L. acidophilus and Bifidobacterium longum was identified after different biochemical analysist which were also showed reliable probiotic properties. The antibacterial activity of lactic acid bacteria (LAB) isolated from raw milk against common enteric pathogens. The antagonistic properties of these isolates against Escherichia coli, Staphylococcus aureus, were examined using agar well diffusion method. Shobha Mehra | Vimla Mehra | Dinesh Bhauryal"Isolation, Identification of Probiotic Bacteria Present in Milk" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-2 | Issue-5 , August 2018, URL: http://www.ijtsrd.com/papers/ijtsrd16979.pdf http://www.ijtsrd.com/biological-science/biotechnology/16979/isolation-identification-of-probiotic-bacteria-present-in-milk/shobha-mehra
MPN is most commonly applied for quality testing of water, i.e., to ensure whether the water is safe or not in terms of the bacteria present in it. The presence of very few faecal coliform bacteria indicates that the water is likely free of disease-causing organisms.
video link: https://youtu.be/vOB9AQo54vU
viable but non-culturable microorganisms (VBNC).
WHAT IS A VBNC STATE?
CHARACTERISTICS OF BACTERIA IN VBNC STATE.
CONDITIONS STIMULATING VBNC STATE.
METHODS FOR DETECTION OF VBNC STATE.
Impact on public health.
Advantages of VBNC.
Disadvantages of VBNC state.
Conclusion.
Very important multiple choice question on Industrial Microbiology
Subscribe this you tube channel for more videos on Microbiology and MCQs
https://www.youtube.com/channel/UCxqXy8R-dkqodPMXnXvE8pQ
this presentation elaborates about the process of producing baker's yeast in detail
contents:1)Introduction
2)media and other raw material preparation
3)fermentation conditions
4)industrial preparation
5)Flowchart for the production of baker’s yeast
6)applications of bakers yeast.
Mutagenesis; A conventional tool for strain improvement in industry Zohaib HUSSAIN
The strain improvement is the process of improvement and manipulation of microbial strains for the icreasment of metabolic level for industrial applications. The yield of microbial enzymes can be increased by using microbe specific medium for fermentation, improving the fermentation process and strain improvement for higher yield of product.
All these things lead to decrease in cost production. Microbe produce product according to its need therefore there is great need for overproduction. There is tremendous contribution of conventional Mutagenesis for strain improvement. Mutagenesis is important tool for the production of mutants which are capable to produce large product i.e. hyperactive.
An overview of lactic acid bacteria - IJBInnspub Net
Lactic acid bacteria (LAB) are renowned for the potential of producing antimicrobial compound and other value added products. Undeniable to concern these probiotic has contributed to the importance of human life. Deserving an attention for its capabilities, this paper will discuss on the general description of lactic acid bacteria, genetics, metabolism and its application to the industries. Get the full articles at: http://www.innspub.net/volume-1-number-3-june-2011/
Very important multiple choice question on Industrial Microbiology
Subscribe this you tube channel for more videos on Microbiology and MCQs
https://www.youtube.com/channel/UCxqXy8R-dkqodPMXnXvE8pQ
this presentation elaborates about the process of producing baker's yeast in detail
contents:1)Introduction
2)media and other raw material preparation
3)fermentation conditions
4)industrial preparation
5)Flowchart for the production of baker’s yeast
6)applications of bakers yeast.
Mutagenesis; A conventional tool for strain improvement in industry Zohaib HUSSAIN
The strain improvement is the process of improvement and manipulation of microbial strains for the icreasment of metabolic level for industrial applications. The yield of microbial enzymes can be increased by using microbe specific medium for fermentation, improving the fermentation process and strain improvement for higher yield of product.
All these things lead to decrease in cost production. Microbe produce product according to its need therefore there is great need for overproduction. There is tremendous contribution of conventional Mutagenesis for strain improvement. Mutagenesis is important tool for the production of mutants which are capable to produce large product i.e. hyperactive.
An overview of lactic acid bacteria - IJBInnspub Net
Lactic acid bacteria (LAB) are renowned for the potential of producing antimicrobial compound and other value added products. Undeniable to concern these probiotic has contributed to the importance of human life. Deserving an attention for its capabilities, this paper will discuss on the general description of lactic acid bacteria, genetics, metabolism and its application to the industries. Get the full articles at: http://www.innspub.net/volume-1-number-3-june-2011/
Storage of Lactic Acid Bacteria Isolated form Honey Bees in Inverted Sugar SyrupAgriculture Journal IJOEAR
Abstract— Honey bee hives are influenced by many different biotic and abiotic factors. Several studies demonstrate that the presence of the lactic acid producing bacteria such as Lactobacillus, Bifidobacterium, and Leuconostoc in honey bee intestinal tracts has a positive effect on a bee hive’s health. Bee hives which are lacking beneficial microflora could be supplied with it through a carrier media. The aim of this study was to evaluate several concentrations of inverted sugar syrup’s suitability for carrying lactic acid bacteria for possible enhancement of honey bee microflora by using lactic bacteria strains previously isolated from honey bee gastrointestinal tract in Latvian apiaries. Since sugar syrup is commonly used for feeding honey bees during the wintering it was decided to use it as a carrier media. There were five isolates of lactic acid bacteria used to test three concentrations of sugar syrup and it was found out that the inverted sugar syrup concentration of 25% is most suitable for carrying tested isolates judged by their growth dynamics and attraction from the bees.
A short presentation on how to create a Lactobacillus culture (LAB) for use in horticulture. This can be a basis for many things from fermentation to odour control - But by far the best is for making your own free organic liquid plant food!!
Microbiology lab report :
1- introduction (Gram Staining)
2-Requirements (Reagents)
3-Method
4-Observations
College of Medicine
Al-Imam Mohammad Bin Saud Islamic University
First year
2013 – 2014
by asem shadid
Lab Report: Isolation of Pure Culture, Gram-staining, and Microscopic Observa...Annisa Hayatunnufus
A Lab Report under the subject of Microbiology. Done as a lab session in Josai University, Japan during a twinning program on 2014.
Created by: Annisa Hayatunnufus
Bachelor of Pharmacy
Management & Science University
Identification of bacteria, Bacterial identification, Lab identification of b...
Similar to ISOLATION AND IDENTIFICATION OF LACTIC ACID BACTERIA FROM PLANTS AND OTHER VEGETABLE MATRICES AND MICROBIAL RECOMBINATION WITH ENTEROCOCCUS SPP.
Preservative potentials of crude bacteriocins produced by Lactobacillus tucce...iosrjce
IOSR Journal of Biotechnology and Biochemistry (IOSR-JBB) covers studies of the chemical processes in living organisms, structure and function of cellular components such as proteins, carbohydrates, lipids, nucleic acids and other biomolecules, chemical properties of important biological molecules, like proteins, in particular the chemistry of enzyme-catalyzed reactions, genetic code (DNA, RNA), protein synthesis, cell membrane transport, and signal transduction. IOSR-JBB is privileged to focus on a wide range of biotechnology as well as high quality articles on genetic engineering, cell and tissue culture technologies, genetics, microbiology, molecular biology, biochemistry, embryology, cell biology, chemical engineering, bioprocess engineering, information technology, biorobotics.
Antimicrobial activity of Lactobacillus spp. especially (L. planetarium and L. acidophilus) against S. aureus were tested using agar-plug, agar well diffusion methods to select the best isolate that could inhibit the growth of multidrug resistance isolates. Further identification for the presence of bacteriocin was done using ELISA kit. Results showed that Lactobacillus spp isolates were bacteriocin producers with different degrees and that L. planetarium (L7) was the most efficient in bacteriocin production. Therefore, L. planetarium (L7) was selected for purification using 70% saturated ammonium sulfate and gel chromatography. The effect of purified bacteriocin was tested on 16 bacterial isolates using micro-titer plate method and well diffusion method. The results showed the ability of the bacteriocin to inhibit bacteria only at concentrations 1866U/ml (50%), 3732U/ml (100%) with a diameter of inhibition zones ranges between (11-23 mm) respectively. The anti-biofilm activity of purified bacteriocin at concentration 100% was investigated and the results showed that biofilm formation was reduced by 100% in the presence of bacteriocin.
Detection of Slime-Producing Staphylococcus aureus Strains Isolated from Food...Agriculture Journal IJOEAR
Abstract— The contamination of food with pathogenic microorganisms producing biofilm, implies a high cost for the food industry and represents a serious risk for the health of consumers. The antibacterial activity of organic extracts of Azorella trifurcata and Mulinum echegarayii was evaluated against 4 Staphylococcus aureus slime-producing strains isolated from bakery foods and against S. aureus ATCC 35556 slime-producing strain and S. aureus ATCC 25923 non slime-producing strain. The plant extracts showed antibacterial effectiveness against all the strains of S. aureus tested with minimum inhibitory concentration (MIC) between 500 and 8000 µg/ml. M. echegarayii 30:70% AcOEt:HEX showed the best activity: five strains of S. aureus showed MIC of 1000 μg/ml and S. aureus ATCC 25923 was inhibited at doses of 500 μg/ml. The values of minimum bactericidal concentration (MBC) of the extracts assayed were one or two times higher than corresponding MIC values. This study showed that extracts of Azorella trifurcata and Mulinum echegarayii are promising for future natural therapy against slime-producing S. aureus. Plant extracts with activity against slime producing S. aureus strains could provide benefits for of food technology and public health.
Similar to ISOLATION AND IDENTIFICATION OF LACTIC ACID BACTERIA FROM PLANTS AND OTHER VEGETABLE MATRICES AND MICROBIAL RECOMBINATION WITH ENTEROCOCCUS SPP. (20)
INDENTURED LABOUR IN THE COLONIAL TEA PALANTATIONS OF ASSAM: A SAGA OF EXPL...American Research Thoughts
Abstract: This article of mine examines the development of indentured labor plantations in the
colonial tea gardens of Assam. The laborers were recruited through licensed recruiters and most of
them came from Chotanagpur region. The notion of the Adivasi coolie having the capacity to perform
arduous tasks was becoming popular. Subsequently, this article examines the exploitation and
sufferings inherent in the system of indenture and how these manifested into forms of collective
violence and large scale desertions in the tea gardens. This is an original work bereft of any sort of
plagiarism whatsoever and has not been sent for publishing anywhere. This study has been based
predominantly upon secondary sources with primary sources in the National Archives of India and
Assam State Archives and a few reports and conference proceedings.
AN UPDATED REVIEW ON THE CONVENTIONAL AND UNCONVENTIONAL RICH PICTURE BUILD...American Research Thoughts
Abstract: The topic of rich picture building exercise (RPBE) comes from the Soft Systems
Methodology (SSM) of P.B. Checkland in the early 80s. Since then, it has been studied and reported
in the academic literature. The recent academic works inspires the writer to review this topic and
argues that the mainstream study on RPBE can be considered as a conventional RPBE. The paper
then points out that there is also an unconventional RPBE which has been neglected in the academic
community. It reasons that the unconventional RPBE is also useful to express the proble m situation,
which is what stage 2 of the conventional SSM is all about. The unconventional RPBE can be
conducted either solely or as a complementary exercise with the conventional RPBE. A number of
RPBE options are identified in the discussion. The paper is intended to contribute to the theoretical
development of the RPBE.
PSYCHO-SOCIAL PROBLEMS OF ELEMENTARY SCHOOL WOMEN TEACHERS IN RELATION TO T...American Research Thoughts
Abstract: Teachers have been increasingly presented with significant occupational health problems.
The psycho-social problem of teachers results from heavy work burden, heavy life pressure, and high
expectation from children’s parents, harassment about interpersonal relationship and the shortage of
their knowledge on psychological health. Positive environment is the basis for psycho-social health, so
creating a stress-free and harmonious environment comes first in order to maintain teac hers’ psychosocial problems. Descriptive survey method was used. The sample for the study was selected as 197
elementary women teachers working in government and private schools by using stratified random
sampling from the districts of Wokha, Kohima and Dimapur of Nagaland.
The results reveal that majority of the elementary school women teachers have average level of psychosocial problems and the elementary schools in Nagaland have controlled and familiar climate. There is
low disengagement, low alienation, moderate esprit, moderate intimacy, low psycho -physical
hindrance, moderate controls, moderate production emphasis and moderate humanized thrust. There
is no significant difference between school organisation climate and psycho -social problems of
elementary school women teachers working in government and private schools in Nagaland. It was
also found out that the women with very high psycho-social problems have high correlation with their
school organization climate.
INDIAN LEGAL AND POLICY FRAMEWORK: WIDENING CRITERION OF WOMEN EMPOWERMENT ...American Research Thoughts
The principal of gender equality is enshrined in the Indian Constitution in its Preamble,
Fundamental Rights, Fundamental Duties and Directive Principles. The Constitution not only grants
equality to women, but also empowers the State to adopt measures of positive discrimination in favour
of women. Women Empowerment is an issue of immense significance today. It is a direct reflection of
the wholesome development of society. Ensuring access to justice for the vulnerable and marginalized
sections of the society is a precondition for a healthy democratic and a peaceful society. In the wake of
some deeply condemnable incidents dominating the headlines these days, this issue has become
indispensable ethically, morally, legally and socially. The Government of India has enacted certain
laws and acts, which protect the rights of women and allow them to seek legal assistance and justice
when faced with certain problems. By showcasing an impressive range of legal empowerment
approaches in diverse geographical and cultural settings, the value of empowering women, through
efforts aimed at legal education , legal training, and the provision of legal services and the creation of
space for women to question and negotiate discriminatory ‘cultural’ norms. The objective of this paper
is to study the laws passed in India for women empowerment i.e. women-specific legislations &
women-related legislations. The paper discuss in details:
Protection of women from sexual harassment at work place
Legal provisions for crimes against women under Indian penal code, 1860.
Provisions leading towards legal empowerment
The paper highlights the critical role legal empowerment strategies that can play in changing
oppressive gender relations that are justified under the name of so called ‘culture’. The paper
concludes with suggesting few provisions which will lead to legal empowerment.
Abstract: The purpose of this paper is the formulation of a framework for assessing development
change in small developing countries modified for application to small islands, and further, to propose
a development process to be used alongside the framework. The methodology utilized in the case study
involved research of official published documents, analysis of relevant statistical data, and application
of Landsat imagery for producing a land cover map. The main finding is that the existing assessment
framework developed by the European Environmental Agency does not fully fit the conditions in
small islands and has to be modified for implementation which is better used in combination with a
development process that provides a better fit for purpose. The modified framework and the detailing of
a new development process presented in this paper are original in the suggested applications and will
be valuable to the agencies that carry the responsibility for undertaking environmental and
development assessments in small countries.
Abstract: Today’s citizens need to think globally while acting locally, conscious of the fact that their
actions can have far reaching implications despite their absence. And teacher’s actions in the
classroom are pertinent to the development of future adult citizens. Human actions are nothing but a
manifestation of their thoughts, teachers being human their actions whether within or outside the
classroom too is a result of their thoughts. Present day schools no more house Monocultural
Traditional Classrooms but have been replaced by Culturally Diversified Smart Classroom where in
the main motto is ‚Sabke Saath, Sabka Vikas‛. Here no one is to be left behind on the basis of
ethnicity, race, socio-economic status, gender, exceptionalities, language, religion, sexual orientation
and geographical area. This demands that teachers impartially identify, tap-in and provide
opportunities for the development of the unique abilities of the students. For this teachers will have to
be able to carry out Higher Order Thinking Skills (HOTS) that involves critical thinking, problem
solving and creating with critical thinking at its foundational level. In fact a 21st century teacher to
fully participate in the Global community needs to Master not only the 3R’s of reading, writing and
arithmetic, but also the 4C’s i.e. critical thinking, creativity, communication and collaboration. The
term ‘Master’ reminds us of the fact that effective teachers are made and not born, hence the concept of
Teacher Education. Teacher Education is a teacher preparation programme wherein one of the main
objectives is to help teachers develop the required skills to perform their task effectively in the school
and classroom, here the Critical Thinking Skills. Here Albert Einstein can be aptly quoted where he
stated: ‚Education is not the learning of facts but training the mind to think‛. The term Critical
Thinking too finds expression in the National Curriculum Framework for Teacher Education 2009-10
which aims at developing teacher’s ability to think and be critical, thus producing Humane and
Professional teachers.
IMPACT OF EDUCATION OF PARENTS ON ADJUSTMENT AND ANXIETY OF STUDENTS OF AFF...American Research Thoughts
Abstract: The research has been done on impact of education of parents on the adjustment and
anxiety of B.Ed. students. The sample consists of 367 B.Ed. teacher trainees of affiliated B.Ed. colleges
in Faridabad and Gurgaon region. Mean standard deviation and t-test is used in the present study.
DESFLORESTAMENTO NO SUL DO ESTADO DO AMAZONAS: EMBATE ENTRE O CRESCIMENTO E...American Research Thoughts
Resumo: O sul do Estado do Amazonas, localizado na Região Norte do Brasil, vem sofrendo com o
crescente desmatamento causado por atividades econômicas oriundas dos estados vizinhos localizados
no Centro-Oeste brasileiro. A pecuária, a produção de grãos e a exploração da madeira têm avançado
para a Região Norte chegando à Sub-Região do Purus trazendo preocupações tanto para o Governo do
Estado quanto para o Governo Federal que procuram neutralizar a dinâmica do desmatamento por
meio de medidas políticas estabelecidas em legislação específica. Porém, o problema encontra
dificuldades de ser equacionado porque o vetor principal deste desmatamento pode estar no próprio
processo de crescimento econômico do país. Este artigo objetiva fazer uma análise parcial do
desmatamento na região, discorrendo sobre os seus impactos no ambiente e nas populações autóctones
e também analisar a falha metodológica de aplicação do Código Florestal Brasileiro, mostrando que do
ponto de vista econômico a imposição de solução uniforme para problemas ambientais de natureza
diversa se caracteriza pela ineficiência dos seus resultados.
Abstract: The southern state of Amazonas, located in the northern region of Brazil, has suffered from
increasing deforestation caused by economic activities originating from neighboring states located in
the Brazilian Midwest. Livestock, grain production and the exploitation of wood have advanced to the
Northern Region reaching the Purus Subregion bringing concerns both the State Government and to
the federal government seeking to neutralize the dynamics of deforestation through policy measures
established in specific legislation. However, the problem is difficult to be solved because the main
vector of this deforestation can be in itself of the country's economic growth. This article aims to make
a partial analysis of deforestation in the region, discussing their impact on the environment and on
indigenous peoples and also examine the methodological failure of application of the Brazilian Forest
Code, showing that from an economic point of view the solution uniform enforcement for problems
environmental diverse in nature is characterized by inefficiency of their results.
FRESHWATER FARMING OF BRACKISHWATER SHRIMP, PENAEUS MONODON (FABRICIUS) WIT...American Research Thoughts
Abstract: Brackish water shrimp (Penaeus monodon) farming expanded rapidly after the technical
viability of this culture system was established and farmers discovered that the high profits derived
from shrimp production could easily offset increased costs associated with this culture. These factors
facilitate the spread of brackish water shrimp farming into freshwater agricultural areas of Purba
Medinipur district of West Bengal that never experience salt water intrusion. The emergence of
brackish water shrimp farming within paddy growing regions of Purba Medinipur district has raised
concerns regarding potential environmental impacts and the suitability of conducting this activity
within highly productive freshwater agricultural areas. In the present study an attempt had been
made to farm the black tiger shrimp, Penaeus monodon in almost freshwater condition with
innovative technologies in 04 earthen tanks each with 0.4 ha water spread area under Contai -III Dev.
Block in Purba Medinipur district in the year 2011 (April to August). The PCR tested P. monodon
seeds (PL15) were stocked in all freshwater earthen tanks after proper acclimatization @
50,000nos/tank. The salinity of the tanks was recorded between 0.0063 ppt to 0.04 ppt. The shrimps
were fed with branded feed and the feeding schedule was based on check-tray method as well as a feed
chart given by the concerned manufacturer.
Abstract: Implementation of Information & Communication Technology has changed t he way people
access and communication information. Users want easy and instant access to relevant information.
So putting pressure on library professionals to think out of the box for meeting their information
needs. Application of mobile phones to provide library and information services are significant step in
this direction. The development of mobile technology has resulted in shifting the academic
environment from traditional to mobile learning setting. This paper high-light the assumption of
mobile technology in library services, its purpose, literature review various devices, advantages,
disadvantages and prerequisites for implementing mobile based services etc.
Abstract: Vorticity is a curl of fluid velocity and the Absolute ξ_a = ξ +f where ξ is the relative
vorticity while f is the coriolis parameter. North word moving air acquires and increasing value for f
and hence decreasing value for ξ. The concept of circulation is related that of vorticity, and it has a
number of important applications horizontal closed curves in either hemisphere will have positive
circulation vorticity is negative while the curvature is negative the vorticity is positive.
GENETIC RESOURCES FAMILY POACEAE IN THE FLORA OF NAKHCHIVAN AUTONOMOUS REPU...American Research Thoughts
Abstract: This article provides information about the current state of taxonomy, genetic resources of
species belonging to the family of cereals - Poaceae Barnhart in flora of Nakhchivan Autonomous
Republic of Azerbaijan. Clarified that this family is represented by 95 genus and 297 species.
Contains: 1-Ordo-Poales, 1-Subfamilya, 13-Triba, 88-Sector, 12-Subgen, 60-Subsp. and 23-Variasya.
32 species are rare-endangered. There are new taxa are endemic and their wild relatives, the protection
of which is considered to be important issues.
Abstract: Numerous indicators of crucial role exist for improvements of the students' performance
during the learning process in classrooms, to include the level of teaching and learning, teachers’
performance, students’ motivation and teaching methods. However, the class size has its impact over
the successful performance of students, which is the main topic of this scientific research. Regarding
the impact of class size, or more precisely, the number of students in the classroom, in student
performance, has developed an empirical research which results are reflected in this paper. The survey
was conducted in several schools of higher education in some Kosovo municipalities. by the outcomes
introduced it is intended to prove a dilemma that has existed for a long time for classes with smaller
number of students which tend to be more successful compared to the classes with higher number of
students. Furthermore, the review of the literature, is an important part of the paper, where intertwine
various theories regarding this issue, bringing different experiences of OECD countries regarding the
impact of class size on student performance. Finally, the performance of the students was tested in
different class sizes at the end of the school year to see if there is a difference in their outcome.
INDENTURED LABOUR IN THE COLONIAL TEA PALANTATIONS OF ASSAM: A SAGA OF EXPL...American Research Thoughts
Abstract: This article of mine examines the development of indentured labor plantations in the
colonial tea gardens of Assam. The laborers were recruited through licensed recruiters and most of
them came from Chotanagpur region. The notion of the Adivasi coolie having the capacity to perform
arduous tasks was becoming popular. Subsequently, this article examines the exploitation and
sufferings inherent in the system of indenture and how these manifested into forms of collective
violence and large scale desertions in the tea gardens. This is an original work bereft of any sort of
plagiarism whatsoever and has not been sent for publishing anywhere. This study has been based
predominantly upon secondary sources with primary sources in the National Archives of India and
Assam State Archives and a few reports and conference proceedings.
DEVELOPMENT AND TESTING OF A MODIFIED 500 GPS SOLAR POWERED POND AERATOR SY...American Research Thoughts
Abstract: One of the problems of fish farming in ponds is the dissolved oxygen (DO) concentration
in water. Lack of dissolved oxygen from just a few feet below the surface of the pond causes fish to
swim on the water surface. Fish may die either from warm water where oxygen becomes less soluble at
higher temperature or from ammonia and carbon dioxide that turned into gas bubbles in their
bloodstream. The best way to control or eliminate the causes of fish kill is to ensure that there is
always enough oxygen in the pond. To minimize, if not to eradicate this problem, a study on the
development and pre-testing of a modified 500 gallons per hour Solar Powered Pond Aerator System
project also known as SPAS-500 was conducted. It is a self-contained unit that floats on the water and
provides consistent levels of oxygen using the venturi air ejector technology. It directly uses the
collected energy from the sun during daytime and the stored energy of the batteries at night time.
The method of the evaluation was based on the data gathered from the hydrological and electrical
application of the system. Varied methods of computations and testing as well as measuring
instruments/devices were utilized to achieve reliable results and significant conclusions. Finally, the
study showed that utilization of the SPAS-500 yielded a significant impact on economic and
environment. It means that the device can address the minimum aeration requirements needed by
small and medium size aquamarine businesses.
Abstract: UGC has splendor activities in respect of conducting of sixteen autonomous statutory
institutions, co-ordination, formulation and maintenance of the standards of university education,
framing regulations on minimum standards of education, Determining standards of teaching,
Examination and research in universities, Monitoring developments in the field of collegiate and
university education, Disbursing grants to universities and colleges, Setting up common facilities,
services and programs for a group of universities in the form of Inter-University Centers but there are
lots of issues (negative performances) are related with UGC’s performance for these National
Commission for Higher Education & Research (NCHER – A national over-arching regulatory body
in higher education), Rashtriya Uchchatar Shiksha Abhiyan (RUSA) and demand of CBSC have been
aroused. This article will highlight the issues related to cause of emerged of alternative body of UGC
in India.
For India to emerge as knowledge super power of the world in the shortest possible time it is
imperative to convert our demographic advantage into knowledge powerhouse by nurturing and
honing our working population into knowledge or knowledge enabled working population.
As multimedia teaching technologies become more widely advocated and employed in
education, researchers strive to understand the influence of such technologies on student learning.
Advances in technology enable pedagogical enhancements that some believe can revolutionize
traditional methods of teaching and learning. When viewed collectively, these studies reported
that advanced technologies, especially multimedia instruction, which often involves introducing or
enhancing the visual aspects of the presentation of course contents, created an active learning
environment, improved students' performance, fostered positive attitudes toward learning
complex concepts, increased communication and could be adapted to all learning styles and levels
of instruction. Researchers suggest that, compared to classes with a traditional teacher-leading
approach, those using multimedia are better liked by students and yield slight but statistically
significant improvements in student learning as measured by both student self-report and
objective outcome testing. Such encouraging findings have precipitated the adoption of these
technologies on a widespread basis. Despite many studies suggesting that, multimedia instruction
benefits students. Therefore, there is a need to further educators' understanding of the effect of
multimedia technologies on students' learning quality.
The combined outcomes of the majority of studies across disciplines indicated that
multimedia-based delivery systems offered ways to optimize the advantages and minimize the
disadvantages of traditional methods of teaching and learning. These are expected to be true in
biology. Biology laboratories are designed to help students understand the basic concepts and their
applications by experiments, collecting specimens, using specimens to know the parts and
functions, and drawing and writing a laboratory report. Many factors such as the time limit for
setting up, the unavailability of specimens in the traditional laboratories. However, the
disadvantages elicited by these factors can be addressed with the use of multimedia-based delivery
systems.
CONSERVATIONAL STUDIES ON CHLOROPHYTUM BORIVILIANUM (SAFED MUSLI) IN NANDUR...American Research Thoughts
An ethno-ecological and ethno-medicinal survey was conducted in various parts of
Nandurbar dist. of Maharashtra to find out the cause of loss of biodiversity and abundance of
Chlorophytum borivilianum. Investigation are proceeds to tracing and finding the alternative ways to
conservation of Chlorophytum borivilianumat their natural as well as to introductions as a crop in
agricultural sector due to huge ethno economic prospective.
EFFECT OF DIFFERENT PHOTO PERIODS ON THE DEVELOPMENT OF CHRYSOPERLA CARNEA ...American Research Thoughts
Experiment on effect of different photo periods on the development of Chrysoperla carnea
(Stephen) with special reference to egg to adult emergence were carried at Agriculture Research
Institute, Tandojam. The Lacewings Chrysoperla carnea eggs, larvae, pupae and adults were obtained
from Biological control laboratory reared under constant temperature 26± 2
o
C with relative humidity
65-70%. Photoperiod significantly affected the development from egg to adult (Table 1). In complete
darkness (L: 0 D: 24), resulted in minimum hatching, larval survival (No. of pupae) and hatching
time, larval period and pupal period was longer significantly as compared to other treatments.
Whereas, the treatment with complete light hours (L: 24, D: 0) resulted in maximum hatching, larval
survival (No. of pupae) and adult emergence (table 1). The hatching time, larval and pupal duration
were significantly shortened. Sex ratios were also female biased when full light hours were provided
for development. The finding of this study it is concluded that Developmental period of C. carnea from
egg to adult was shorter at 24:0 because mass rearing predator C. carnea in laboratory required
shorter time to complete life duration, and we will get more number of biocontrol agents for release in
field.
June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
• The Committee on Ways and Means has been investigating several universities since November 15, 2023, when the Committee held a hearing entitled From Ivory Towers to Dark Corners: Investigating the Nexus Between Antisemitism, Tax-Exempt Universities, and Terror Financing. The Committee followed the hearing with letters to those institutions on January 10, 202
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Read| The latest issue of The Challenger is here! We are thrilled to announce that our school paper has qualified for the NATIONAL SCHOOLS PRESS CONFERENCE (NSPC) 2024. Thank you for your unwavering support and trust. Dive into the stories that made us stand out!
Embracing GenAI - A Strategic ImperativePeter Windle
Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
A Strategic Approach: GenAI in EducationPeter Windle
Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
Digital Tools and AI for Teaching Learning and Research
ISOLATION AND IDENTIFICATION OF LACTIC ACID BACTERIA FROM PLANTS AND OTHER VEGETABLE MATRICES AND MICROBIAL RECOMBINATION WITH ENTEROCOCCUS SPP.
1. 1503
AMERICAN RESEARCH THOUGHTS
ISSN: 2392 – 876X
Available online at: www.researchthoughts.us
http://dx.doi.org/10.6084/m9.figshare.1425170
Volume 1 │ Issue 7 │ May 2015
Impact Factor: 2.0178 (UIF)
ISOLATION AND IDENTIFICATION OF LACTIC
ACID BACTERIA FROM PLANTS AND OTHER
VEGETABLE MATRICES AND MICROBIAL
RECOMBINATION WITH ENTEROCOCCUS SPP.
Immacolata Anacarsoi
, Lara Bassoli, Carla Sabia, Ramona Iseppi,
Carla Condò
Dept. Life Sciences, University of Modena and Reggio Emilia, Italy
Abstract: Twenty-two lactic acid bacteria (LAB) belonging to Lactobacillus, Lactococcus and
Enterococcus genera were isolated from plants, flowers and other vegetable matrices. The predominant
LAB species were Lactobacillus brevis (57%) followed to Lactobacillus delbrueckii subsp. bulgaricus
(14%) and Lactococcus lactis (14%). Among enterococci E. faecalis (4 /50%), E. faecium (2 /25%), E.
hirae (2 /12,5%) were isolated. The strains were identified previously with biochemical kits and then
confirmed by PCR. Bacteria isolated were tested against nine strains selected among pathogenic and
opportunistic strains to evaluate their capacity to produce BLS (bacteriocin like substance).
Subsequently to demonstrate the horizontal transfer of genes coding for the production of bacteriocin
substances, a conjugation experiment was positively carried out between an Enterococcus faecium
used as donor and a Lactobacillus acidophilus used as recipient.
Key Words: Lactobacillus, Enterococcus, conjugation, bacteriocin.
INTRODUCTION
The lactic acid bacteria (LAB) are generally defined as a cluster of lactic acid-producing,
low %G + C, non-spore-forming, Gram-positive rods and cocci, catalase negative
bacteria which share many biochemical, physiological, and genetic properties. LAB
i Dr. Immacolata Anacarso – Dept. Life Sciences University of Modena and Reggio Emilia
Via Campi, 287 – 41125 - Modena (Italy)
Tel. +39 0592055795, Fax +39 0592055483.
E-mail: immacolata.anacarso@unimore.it
2. Immacolata Anacarso, Lara Bassoli, Carla Sabia, Ramona Iseppi, Carla Condò- ISOLATION AND
IDENTIFICATION OF LACTIC ACID BACTERIA FROM PLANTS AND OTHER VEGETABLE MATRICES
AND MICROBIAL RECOMBINATION WITH ENTEROCOCCUS SPP.
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group include the most important groups of microorganisms like Lactobacillus,
Lactococcus, Enterococcus, usually used in food fermentations. These microorganisms in
fact contribute to the taste and to the texture of fermented products and inhibit the food
spoilage and pathogenic bacteria by producing an array of antimicrobial substances
such as large amounts of lactic acid (and other organic acids), hydrogen peroxide,
antifungal peptides, and bacteriocins (Daeschel, 1989; Dobrogosz and Lindgren, 1990).
Bacteriocins are the best-known antimicrobial compounds isolated from LAB.
These antimicrobial compounds show bactericidal activity against species closely
related to producers (Paker et al., 1989; Cintas et al., 2001) and they can exhibit
heterogeneous characteristics such as resistance to very low pH values, stability at a
very wide range of temperatures, resistance to organic acids, salts and enzymes. Some
studies show the potential use of bacteriocins as food preservatives or additives to
packaging for extend the products shelf life.
The importance of the LAB is because these bacteria have beneficial effects to the
consumers in different ways. Probiotic lactobacilli, for example, are known to confer an
array of health promoting activities on their host after either parenteral or oral
administration (Oyetayo and Osho, 2004). The other beneficial effects of the LAB
include prevention of intestinal infections (Casas and Dobrogosz, 2000),
anticarcinogenic activity (Kumari et al., 2011), control of serum cholesterol, immunity
promotion (Aattouri et al., 2001), and growth enhancement of animals (Chang et al.,
2001). The mechanism by which these probiotics affect their host and bring an
improvement in the gut barrier can be thanks to competition for the adhesion site, the
production of inhibitory compounds, the rebalancing of disturbed gastrointestinal
microbial composition and of the metabolism (FAO/WHO, 2001). Many are the study
about the new isolated LAB capable to produce natural bacteriocin substances, with a
broad spectrum. Many times the best bacteriocins producers belonging to the
Enterococcus genera. Though enterococci are LAB, and are the natural flora of some
fermented foods (cheese, salami etc.) they are however opportunistic bacteria capable to
produce many virulence factors and able to cause important pathologies (Sabia et al.,
2002); for these reasons the use of enterococci or bacteriocins produced from these
bacteria not always is well accepted. Always actual is the research of new bacteriocin
substances product by LAB as Lactobacillus or Lactococcus. Different are the matrices
from which these LAB were isolated, like kefir grains (Leite et al., 2015), fermented
3. Immacolata Anacarso, Lara Bassoli, Carla Sabia, Ramona Iseppi, Carla Condò- ISOLATION AND
IDENTIFICATION OF LACTIC ACID BACTERIA FROM PLANTS AND OTHER VEGETABLE MATRICES
AND MICROBIAL RECOMBINATION WITH ENTEROCOCCUS SPP.
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sausages (Golneshin et al., 2015), wine (Ndlovu et al., 2015), olive (Hikmate et al., 2015)
usually starting to a fermented substrates.
In this study we have isolated different bacterial strains from plants, flowers and
other vegetable matrices, showing that also in these sample it was possible to isolate
LAB as Lactobacillus and Lactococcus plus Enterococcus. The strains isolated were
interesting because in large part were good BLS (bacteriocins like substances)
producers. The study here presented has also shown a conjugation experiment between
an Enterococcus donor and a Lactobacillus recipient to obtain a best bacteriocins
producers with an broad antimicrobial activity typically of a Enterococcus but from
Lactobacillus producers.
MATERIAL AND METHODS
Sample analysis
Forty different vegetable matrices derived from houseplants (including cacti, potted
flowers, bushes flowers, climber plants, seeds), leaves and flowers trees, were subjected
to microbiological analysis for the research of LAB.
Samples were weighted and diluted in a 1:10 ratio with physiological solution
and then homogenized by Stomacher for 1 minute. The obtained solutions, after
appropriate dilutions in physiological solution, were plated in Kanamycin Aesculin
Azide agar and MRS agar (Biomerieux, Milan-Italy) and incubated at 30°C in aerobic
and anaerobic conditions respectively for 48-72 h.
Bacterial identifications
For a preliminary recognition of isolated strains, several preliminary tests like Gram
stain, microscopic view and catalase analysis were carried out.
All strains grown on MRS agar that were Gram-positive, catalase negative and
cocco- or rod-shaped were before identified with API 50 CHL (Biomerieux, Milan-Italy)
and lastly identified by PCR. All strains grown on Kanamycin Aesculin Azide agar with
a typical aspect for enterococci were before identified with API 20 Strep (Biomerieux,
Milan-Italy) and lastly identified by PCR. DNA was extracted according to the method
of Marzotto et al., (2006). The PCR reactions were carried out by Techne Termal cycler
TC312 (Bibby Scientific, Italy). The following conditions were used: 2 min at 94°C, 30
cycles of 1 min at 92°C, 1 min at optimum temperature based on primers used, 1 min at
4. Immacolata Anacarso, Lara Bassoli, Carla Sabia, Ramona Iseppi, Carla Condò- ISOLATION AND
IDENTIFICATION OF LACTIC ACID BACTERIA FROM PLANTS AND OTHER VEGETABLE MATRICES
AND MICROBIAL RECOMBINATION WITH ENTEROCOCCUS SPP.
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72°C and finally 10 min at 72°C. PCR primers and conditions used are listed in the Table
1. Also, negative controls containing no DNA template were included in the
experiment. All PCR products were electrophoresed in a 1% (w/v) agarose gel with 1 X
Tris/borate/EDTA buffer and viewed under UV light.
Evaluation of BLS (Bacteriocins like Substances) production
BLS production was performed by using the agar-spot-test method (Todorov and Dicks,
2005) on MRS (Man Rogosa Sharp) (Oxoid, Italy) for the LAB, correcting the cell-free
supernatant to pH 6.0 with 1M NaOH to prevent the inhibitory effect of lactic acid, and
on TSA (tryptic soy agar) (Oxoid, Italy) for enterococci. Nine strains were used as
indicators selected among pathogenic and opportunistic strains from registered
collections (ATCC - American Type Culture Collection; NCTC - National Collection of
Type Cultures): Listeria monocytogenes ATCC 13932, Enterococcus casseliflavus 416/K1
(Sabia et al., 2002), Listeria monocytogenes NCTC 10888, Staphylococcus aureus ATCC 6538,
Enterococcus faecalis ATCC 29212, Bacillus subtilis ATCC 6633, Candida albicans ATCC
10231, Escherichia coli ATCC 8739, Pseudomonas aeruginosa ATCC 27853.
Conjugation experiment
This experiment was performed with two of the strains isolated to demonstrate the
horizontal transfer of genes coding for the production of bacteriocins. Twelve different
conjugation experiments to obtain a positive result were done.
A strain of Enterococcus faecium EN313 (KanamycinR, TeicoplaninS), was selected
as donor, because producer of a bacteriocin with high antibacterial activity against
Listeria monocytogenes NCTC 10888, that was used as indicator. A strain of Lactobacillus
acidophilus LB813 (KanamicynS
, TeicoplaninR
), was selected as recipient, totally
incapable to produce any antibacterial substances vs Listeria monocytogenes NCTC 10888.
Overnight MRS broths of both strains were then combined in a ratio of 1:2 and
maintained under agitation for 48-72 h at 30°C and anaerobic conditions.
Conjugation broths were filtered on cellulose acetate filters with 0.45 µM pores
(Millipore, Italy), which were laid on MRS agar with the selective agents (Kanamycin 64
mg ml-1 and Teicoplanin 32 mg ml-1) and incubated at 30°C for 24h under anaerobic
conditions.
The donor and recipient were subjected to plasmid-DNA extraction by
O’Sullivan-Klaenhammer method (1993), to evaluate the presence of plasmids; the
5. Immacolata Anacarso, Lara Bassoli, Carla Sabia, Ramona Iseppi, Carla Condò- ISOLATION AND
IDENTIFICATION OF LACTIC ACID BACTERIA FROM PLANTS AND OTHER VEGETABLE MATRICES
AND MICROBIAL RECOMBINATION WITH ENTEROCOCCUS SPP.
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extracts were electrophoresed in 0.7% (w/v) agarose gel with 1% Tris/borate/EDTA
buffer at 100 mV for 35 min and viewed under UV light.
Isolation of transconjugant
The colonies grown on filter surface were collected, suspended in 1 ml of MRS broth
and then aliquots of 100 µl were plated on MRS agar plates with the addition of both
selective agents (Kanamycin 64 mg ml-1 and Teicoplanin 32 mg ml-1). Plates were
incubated for 48-72 h at 30°C under anaerobic conditions and the bacteria colonies
grown after the incubation were used for a subsequent Gram stain and morphological
viewing under the microscope. Those with a rod-shape (easily distinguishable from the
cocco-shape of the donor strain) were isolated and extracted by O’Sullivan-
Klaehnammer method to evaluate the plasmidic bands. To confirm the belonging of
trasconjugats at the Lactobacillus specie, PCR with L. acidophilus primers listed in the
Table 1 was performed. Lastly, to exclude the contamination by E. faecium a PCR for the
research of a specie-specific ligase ddl E. faecium was performed (Dutka-Malen and al.,
1995). Transconjugant strains were tested by agar-spot-test method; previously
indicated, using L. monocytogenes NCTC 10888 as indicator, for evaluate the presence of
antimicrobial activity against the indicator.
RESULTS
Bacterial identification
Only from 6 vegetable matrices (Pteridium aquilinum leaves, Linum usitatissimum seeds,
Daucus carota root, Ficus benjamin leaves, Aloe Barbadensis leaves, Prunus cerasifera
flowers), of the 40 tested it was possible to isolate 14 different LAB including
Lactobacillus and Lactococcus and 8 different enterococci. The strains identification was
made in first time using API kits and lastly by PCR analysis, confirming the biochemical
results. For the LAB strains, 8 (57%) Lactobacillus brevis, 2 (14%) Lactobacillus delbrueckii
subsp. bulgaricus, 2 (14%) Lactococcus lactis, 1 (7%) Lactobacillus acidophilus and 1 (7%)
Leuconostoc were recovered. For the strains grown with the typical enterococci aspect,
on Kanamycin Aesculin Azide agar, 4 (50%) E. faecalis, 2 (25%) E. faecium, 2 (12,5%) E.
hirae were identified.
In Table 2 are listed the bacterial identifications and vegetable matrices from
which they derived.
6. Immacolata Anacarso, Lara Bassoli, Carla Sabia, Ramona Iseppi, Carla Condò- ISOLATION AND
IDENTIFICATION OF LACTIC ACID BACTERIA FROM PLANTS AND OTHER VEGETABLE MATRICES
AND MICROBIAL RECOMBINATION WITH ENTEROCOCCUS SPP.
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AMERICAN RESEARCH THOUGHTS- Volume 1 │ Issue 7 │2015
Evaluation of BLS (Bacteriocin like Substance) production
The strains were tested for BLS production by agar spot method earlier indicated, using
9 different indicators. Most of the LAB strains showed an antibacterial activity against
L. monocytogenes NCTC 10888, and 3 strains in particular (L. lactis LC214, L. brevis
LB1013, L. brevis LB1113 demonstrated a broad antimicrobial activity against the
majority of the indicator strains. Regarding the enterococci only one not showed
antimicrobial activity vs L. monocytogenes NCTC 10888 and 2 in particular EN313,
EN317 showed a broad antimicrobial activity (Table 3).
Conjugation experiment
Out of 12 matings performed, the successful transfer was obtained in one conjugation,
subsequently confirmed, between Enterococcus faecium EN313 (KanamycinR
,
TeicoplaninS
), used as donor and Lactobacillus acidophilus LB813 (KanamicynS
,
TeicoplaninR
), used as recipient. The plasmid-DNA analysis of parental strains showed
a high weight molecular plasmid for E. faecium EN313 and none plasmid line for
Lactobacillus acidophilus LB813 (data not shown).
Isolation of transconjugants
Transconjugant strains with the following characteristics: rod-shaped, KanamicinR
and
TeicoplaninR with a high antimicrobial activity against L. monocytogenes NCTC 10888,
were isolated. The plasmid-DNA analysis of the trasconjugants showed the passage of
plasmid from donor to recipient. The PCR results confirmed the belonging of
transconjugant strains to the L. acidophlilus species. None amplifications was performed
with specie-specific ligase ddl E. faecium, excluding the enterococci cells presence as
contamination and confirming definitively that the conjugation experiment was
successfully performed.
DISCUSSION
Lactobacilli and other bacteria close related have high potentialities. Many could be
their applications in food as well as in medical fields, remembering that the
effectiveness of LAB is strain-dependent. For all these reasons, new strains are always
researched with characteristics that can help the human health, searching the new
desirable strains in natural niches as plants, foods, fermented products, animals and
7. Immacolata Anacarso, Lara Bassoli, Carla Sabia, Ramona Iseppi, Carla Condò- ISOLATION AND
IDENTIFICATION OF LACTIC ACID BACTERIA FROM PLANTS AND OTHER VEGETABLE MATRICES
AND MICROBIAL RECOMBINATION WITH ENTEROCOCCUS SPP.
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AMERICAN RESEARCH THOUGHTS- Volume 1 │ Issue 7 │2015
humans that constitute natural ecological systems and good sources for LAB. The
beneficial characteristics may be also obtained by genetic manipulation for example to
improve the pH resistance or the antibacterial capacity, tied to capacity to inhibit the
pathogens, in particular in the human gut, that is a typical characteristic recognized for
many LAB. In this study were isolated some LAB, including bacteria belonging to
Lactobacillus, Lactococcus and Enterococcus genera, from different plants or other
vegetable matrices. In particular from 40 vegetable matrices only from 6 of these
(Pteridium aquilinum leaves, Linum usitatissimum seeds, Daucus carota root, Ficus benjamin
leaves, Aloe Barbadensis leaves, Prunus cerasifera flowers), it was possible to isolate 14
different LAB including Lactobacillus and Lactococcus and 8 different enterococci.
Plants may be a good source of LAB but as Stirling and Whittenburg (1963)
suggest, the LAB are not usually part of the normal microflora of the growing plant
indicating the role of insects in the spread of these organisms. Other studies reported
the occurrence of LAB on leaving or decayed plants (Mundt and Hammer, 1968; Mundt
et al., 1969) but not so recent, as well as part of the accumulated information about the
occurrence of lactobacilli (and other LAB members) on plants is derived from
microbiological studies of the fermentation process. The microbial population of LAB is
known in several vegetable products (cabbage, silage raw materials, carrots and beets,
olives and fruits such as grapes and pears, etc.). However, scarce information about the
occurrence of LAB on flowers and other parts of plants is available in the literature.
Moreover, LAB are well known for their antagonism towards other Gram-positive
bacteria, especially taxonomically related species (Listeria spp., Bacillus spp. Micrococcus
spp., etc.), but also against Escherichia coli, Salmonella spp., Helicobacter pylori and
Pseudomonas aeruginosa. It is well known that the presence of lactobacilli is important for
maintenance of the intestinal microbial ecosystem, thanks the inhibitory activity against
pathogenic bacteria. This inhibition could be due to the production of inhibitory
compounds such as organic acids, hydrogen peroxide, and bacteriocins (Todorov and
Dicks, 2005; Kumari et al., 2011; Anacarso et al., 2014).
The results reported here indicate that 20 LAB were isolated and most of these
strains have shown an antibacterial activity against L. monocytogenes NCTC 10888 and
other eight pathogenic/opportunistic bacteria, including the Gram-negative indicator
strains. Inhibition caused by hydrogen peroxide and organic acids was ruled out, in fact
the producer strains were cultured anaerobically and the culture supernatants were
neutralized (pH 6.0) before assaying the antimicrobial activity. These results are in
8. Immacolata Anacarso, Lara Bassoli, Carla Sabia, Ramona Iseppi, Carla Condò- ISOLATION AND
IDENTIFICATION OF LACTIC ACID BACTERIA FROM PLANTS AND OTHER VEGETABLE MATRICES
AND MICROBIAL RECOMBINATION WITH ENTEROCOCCUS SPP.
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accordance with earlier results reported by Trias et al. (2008), who showed that most
LAB originating from fruits and other vegetables displayed good antagonistic activity
against foodborne pathogens, such as, Listeria monocytogenes, Salmonella typhimurium
and Escherichia coli.
In this study was also performed a conjugation to pass the antimicrobial activity
showed against L. monocytogenes from a donor strain, Enterococcus faecium EN313 and a
recipient, Lactobacillus acidophilus LB813. Enterococcus spp. is the most controversial
genus of LAB group and if these bacteria are frequently isolated from several fermented
foods, they are always seen with fear owing to their potential risk in human health
(Klein et al., 1998). In many studies, different enterococci are great bacteriocin
producers with a high activity against pathogenic bacteria as Listeria or others. In our
conjugation experiment it was formed a strain with a high antibacterial activity against
L. monocytogenes typical of a Enterococcus strain but produced from a L. acidophilus,
eliminating the apprehension dated to potential risk for human health of the
enterococci. The positive result of this experiment was displayed by a plasmidic band in
trasconjugant and confirmed with a PCR reaction where the absence of a specie-specific
ligase ddl E. faecium it excluded the enterococci cells presence as contamination. Our
results are of interest considering the important probiotic characteristics of lactobacilli
and their possible use, in particular during antimicrobial therapy and more interesting
is the possibility of to have a probiotic capable to produce a bacteriocin with a high
antimicrobial activity against the main food pathogenic bacteria.
Table 1: Primer sequences used for the identifications
Specie
product
size
(pb)
oligonucleotide
primer
sequence ( 5’-3’) Tm (°C)
Leuconostoc 976 LeuF / LeuR
CGA AAG GTG CTT GCA CCT TTC
AAG
TTT GTC TCC GAA GAG AAC A
55
L. brevis 1340
BREVDIR /
BREVREV
CTT GCA CTG ATT TTA ACA
CCC ACT GCT GGG CGG TGT GTA
CAA GGC
40
L. delbrueckii 450 LDEL7 / Lac2
ACA GAT GGA TGG AGA GCA GA
CCT CTT CGC TCG CCG CTA CT
50
9. Immacolata Anacarso, Lara Bassoli, Carla Sabia, Ramona Iseppi, Carla Condò- ISOLATION AND
IDENTIFICATION OF LACTIC ACID BACTERIA FROM PLANTS AND OTHER VEGETABLE MATRICES
AND MICROBIAL RECOMBINATION WITH ENTEROCOCCUS SPP.
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L. acidophilus 1000 Acido / Lac
TGA ACC AAC AGA TTC ACT TC
TGA CGA CAG CCA TGC ACC A 45
L. lactis 380 G1 / L1
GAA GTC GTA ACA AGG
CAA GGC ATC CAC CGT 40
E. faecium 941 E1 / E2
ATCAAGTACAGTTAGTCT
ACGATTCAAAGCTAACTG 45
E. faecium 190 Efm1 / Efm2
TKCAGCAATTGAGAAATAC
CTTCTTTTATTTCTCCTGTA 50
E. faecalis 209 Efs1 / Efs2
CTGTAGAAGACCTAATTTCA
CAGCTGTTTTGAAAGCAG 50
E. hirae 263 Eh1 / Eh2
AAACAATCGAAGAACTACTT
TAAATTCTTCCTTAAATGTTG 50
Table 2: Bacterial Identifications and vegetable matrices of origins
Matrix
Strain identification
LAB
enterococci
Pteridium aquilinum LB113 - L. brevis EN313 - E. faecium
Linum usitatissimum
LB213 - L. delbrueckii
LB313 - L. brevis
LB413 - L. brevis
Daucus carota
LC114 - L. lactis
LC214 - L. lactis
LC314 - Leuconostoc
EN315 - E. faecium
EN316 - E. hirae
Ficus benjamin LB513 - L. brevis
EN314 - E. faecalis
EN319 - E. faecalis
EN320 - E. hirae
Aloe Barbadensis
LB613 - L. brevis
LB713 - L. brevis
10. Immacolata Anacarso, Lara Bassoli, Carla Sabia, Ramona Iseppi, Carla Condò- ISOLATION AND
IDENTIFICATION OF LACTIC ACID BACTERIA FROM PLANTS AND OTHER VEGETABLE MATRICES
AND MICROBIAL RECOMBINATION WITH ENTEROCOCCUS SPP.
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LB813 - L. acidophilus
Prunus cerasifera
LB913 - L. delbrueckii
LB1013 - L. brevis
LB1113 - L. brevis
EN317 - E. faecalis
EN318 - E. faecalis
Table 3: Antibacterial activity of isolated LAB
Indicators
L. i. E. c. 1 C. a E. c. 2 S. a. E. f. L. m. P. a. B. s.
LB113 ++ - - - - - - - -
LB213 ++ - - - - - +++ - -
LB313 ++ - - - - - +++ - -
LB413 + - - - - + - - -
LB513 + - - - - - - - +
LB613 + - - - - - - - +
LB713 + - - - - - + - -
LB813 - - - - - - - - -
LB913 + - - - - - - - +
LB1013 +++ + - + + + +++ + +
LB1113 + + - + + + ++ + -
LC114 ++ - - - - - +++ - -
LC214 ++ + - + ++ + ++ + +
LC314 ++ - - - - - ++ - -
EN313 ++ - - + + - ++ - +
EN314 + - - - - - + - -
EN315 + - - - - - - + -
EN316 + - - - - - + - -
EN317 +++ - - + + - +++ - ++
EN318 + - - - - - + - -
EN319 ++ - - - - - + - +
EN320 + - - - - - + - -
L .i.: L. ivanovii ATCC 13932; E. c. 1: E. casseliflavus 416/k1; C. a.: C. albicans ATCC 10231;
E. c. 2: E. coli ATCC 6538; E. f.: E. faecalis ATCC 29212; L. m.: L. monocytogenes NCTC 10888;
P. a.: P. aeruginosa ATCC 27853; B. s.: B.subtilis ATCC 6633
11. Immacolata Anacarso, Lara Bassoli, Carla Sabia, Ramona Iseppi, Carla Condò- ISOLATION AND
IDENTIFICATION OF LACTIC ACID BACTERIA FROM PLANTS AND OTHER VEGETABLE MATRICES
AND MICROBIAL RECOMBINATION WITH ENTEROCOCCUS SPP.
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AND MICROBIAL RECOMBINATION WITH ENTEROCOCCUS SPP.
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AND MICROBIAL RECOMBINATION WITH ENTEROCOCCUS SPP.
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