Search results for: endemic strains

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

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

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

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Authors: Mohammed H. Abu-Dieyeh, Mohammad M. Zalloum

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Plant growth-promoting rhizobacteria (PGPR) are known to enhance plant growth and/or reduce plant damage due to soil-borne pathogens. Tomato is the highest consumable vegetable world-wide including Jordan. Fusarium oxysporum is a pathogen that causes well-known damages and losses to many vegetable crops including tomato. In this study, purification of 112 isolates of PGPR strains from rhizosphere environment of different regions in Jordan was accomplished. All bacterial isolates were In-vitro screened for antagonistic effects against F. oxysporum. The eleven most effective isolates that caused 30%-50% in-vitro growth reduction of F. oxysporum were selected. 8 out of 11 of these isolates were collected from Al-Halabat (arid-land). 7 isolates of Al-Halabat exerted 40-54% In-vitro growth reduction of F. oxysporum. Four-week-old seedlings of tomato cultivar (Anjara, the most susceptible indigenous cultivar to F. oxysporum) treated with PGPR5 (Bacillus amyloliquefaciens), and exposed to F. oxysporum, showed no disease symptoms and no significant changes in biomasses or chlorophyll contents indicating a non-direct mechanism of action of PGPR on tomato plants. However PGPR3 (Bacillus sp.), PGPR4 (Bacillus cereus), and PGPR38 (Paenibacillus sp.) treated plants or PGPR treated and exposed to F. oxysporum showed a significant increasing growth of shoot and root biomasses as well as chlorophyll contents of leaves compared to control untreated plants or plants exposed to the fungus without PGPR treatment. A significant increase in number of flowers per plant was also recorded in all PGPR treated plants. The characterization of rhizobacterial strains were accomplished using 16S rRNA gene sequence analysis in addition to microscopic characterization. Further research is necessary to explore the potentiality of other collected PGPR isolates on tomato plants in addition to investigate the efficacy of the identified isolates on other plant pathogens and then finding a proper and effective methods of formulation and application of the successful isolates on selected crops.

Keywords: antagonism, arid land, growth promoting, rhizobacteria, tomato

Procedia PDF Downloads 372

Authors: Yassir Adam Shuaib, Stefan Niemann, Eltahir Awad Khalil, Ulrich Schaible, Lothar Heinz Wieler, Mohammed Ahmed Bakhiet, Abbashar Osman Mohammed, Mohamed Abdelsalam Abdalla, Elvira Richter

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Tuberculosis (TB) is a chronic bacterial disease of humans and animals and it is characterized by the progressive development of specific granulomatous tubercle lesions in affected tissues. In a six-month study, from June to November 2014, a total of 2,304 carcasses of cattle, camel, sheep, and goats slaughtered at East and West Gaash slaughterhouses, Kassala, were investigated during postmortem, in parallel, 101 sputum samples from TB suspected patients at Kassala and El-Gadarif Teaching Hospitals were collected in order to investigate tuberculosis in animals and humans. Only 0.1% carcasses were found with suspected TB lesions in the liver and lung and peritoneal cavity of two sheep and no tuberculous lesions were found in the carcasses of cattle, goats or camels. All samples, tissue lesions and sputum, were decontaminated by the NALC-NaOH method and cultured for mycobacterial growth at the NRZ for Mycobacteria, Research Center Borstel, Germany. Genotyping and molecular characterization of the grown strains were done by line probe assay (GenoType CM and MTBC) and 16S rDNA, rpoB gene, and ITS sequencing, spoligotyping, MIRU-VNTR typing and next generation sequencing (NGS). Culture of the specimens revealed growth of organisms from 81.6% of all samples. Mycobacterium tuberculosis (76.2%), M. intracellulare (14.2%), mixed infection with M. tuberculosis and M. intracellulare (6.0%) and mixed infection with M. tuberculosis and M. fortuitum and with M. intracellulare and unknown species (1.2%) were detected in the sputum samples and unknown species (1.2%) were detected in the samples of one of the animals tissues. From the 69 M. tuberculosis strains, 25 (36.2%) were showing either mono-drug-resistant or multi-drug-resistant or poly-drug-resistant but none was extensively drug-resistant. In conclusion, the prevalence of TB in animals was very low while in humans M. tuberculosis-Delhi/CAS lineage was responsible for most cases and there was an evidence of MDR transmission and acquisition.

Keywords: animal, human, slaughterhouse, Sudan, tuberculosis

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Authors: Gagandeep Singh Grover, Vini Mahajan, Bhagmal, Priti Thaware, Jaspreet Takkar

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Dengue is a mosquito-borne viral disease endemic in many countries in the tropics and sub-tropics. The state of Punjab in India shows cyclical and seasonal variation in dengue cases. The Case Fatality Rate of Dengue has ranged from 0.6 to 1.0 in the past years. The department has initiated a review of the cases that have died due to dengue in order to know the exact cause of the death in a case of dengue. The study has been undertaken to know the other associated co-morbidities and factors causing death in a case of dengue. The study used the predesigned proforma on which the records (medical and Lab) were recorded and reviewed by the expert committee of the doctors. This study has revealed that cases of dengue having co-morbidities have a longer stay in the hospital. Fluid overload and co-morbidities have been found as major factors leading to death, however, in a confirmed case of dengue hepatorenal shutdown was found to be a major cause of mortality. The data obtained will help in sensitizing the treating physicians in order to decrease the mortality due to dengue in future.

Keywords: dengue, death, morbidities, DHF, DSS

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Authors: Aicha Beya Mammeria, Idir Bitam

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Our study focuses on the distribution of the white stork "Ciconia ciconia L. 1758" in the wetlands of El Tarf (North eastern of Algeria): recognized by its remarkable number of breeding pairs, monitoring of nesting, using a GPS has been performed in an attempt to explain the functioning of populations and population strategies for an overall design of its distribution, which has not so far been investigated in this region. Between 2012 and 2013, the number of breeding pairs has increased considerably from 174 in 1996 to 475 in 2007 and 968 in 2013. It should be noted that in the distribution of breeding pairs between 1996 and 2011, there is a significant development since the density of nests increased from 25.22 in 1996 to 84.16 couples/100 km² in 2013. More endemic bread apears in the region, this fluctuation is related to climatic change and changing season. Changes related to local climatic conditions might induce binding conditions for the development of this species.

Keywords: white stork, Ciconia ciconia, wetland El Tarf, northeast Algeria, climatic changing, density

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Authors: Lachin Mikjtarnejad, Mohsen Farzaneh

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Various microbes, such as fungi and bacteria species, are naturally found in the fruit microbiota, and some of them act as a pathogen and result in fruit rot. Among non-pathogenic microbes, yeasts (single-celled microorganisms belonging to the fungi kingdom) can colonize fruit tissues and interact with them without causing any damage to them. Although yeasts are part of the plant microbiota, there is little information about their interactions with plants in comparison with bacteria and filamentous fungi. According to several existing studies, some yeasts can colonize different plant species and have the biological control ability to suppress some of the plant pathogens. It means those specific yeast-colonized plants are more resistant to some plant pathogens. The major objective of the present investigation is to isolate yeast strains from apple fruit and screen their ability to control Penicillium expansum, the causal agent of blue mold of fruits. In the present study, psychrotrophic and epiphytic yeasts were isolated from apple fruits that were stored at low temperatures (0–1°C). Totally, 42 yeast isolates were obtained and identified by molecular analysis based on genomic sequences of the D1/D2 and ITS1/ITS4 regions of their rDNA. All isolated yeasts were primarily screened by' in vitro dual culture assay against P. expansum by measuring the fungus' relative growth inhibition after 10 days of incubation. The results showed that the mycelial growth of P. expansum was reduced between 41–53% when challenged by promising yeast strains. The isolates with the strongest antagonistic activity belonged to Metschnikowia pulcherrima A13, Rhodotorula mucilaginosa A41, Leucosporidium Scottii A26, Aureobasidium pullulans A19, Pichia guilliermondii A32, Cryptococcus flavescents A25, and Pichia kluyveri A40. The results of seven superior isolates to inhibit blue mold decay on fruit showed that isolates A. pullulans A19, L. scottii A26, and Pi. guilliermondii A32 could significantly reduce the fruit rot and decay with 26 mm, 22 mm and 20 mm zone diameter, respectively, compared to the control sample with 43 mm. Our results show Pi. guilliermondii strain A13 was the most effective yeast isolates in inhibiting P. expansum on apple fruits. In addition, various biological control mechanisms of promising biological isolates against blue mold have been evaluated to date, including competition for nutrients and space, production of volatile metabolites, reduction of spore germination, production of siderophores and production of extracellular lytic enzymes such as chitinase and β-1,3-glucanase. However, the competition for nutrients and the ability to inhibit P. expansum spore growth have been introduced as the prevailing mechanisms among them. Accordingly, in our study, isolates A13, A41, A40, A25, A32, A19 and A26 inhibited the germination of P. expansum, whereas isolates A13 and A19 were the strongest inhibitors of P. expansum mycelia growth, causing 89.13% and 81.75 % reduction in the mycelial surface, respectively. All the promising isolates produced chitinase and β-1,3-glucanase after 3, 5 and 7 days of cultivation. Finally, based on our findings, we are proposing that, Pi. guilliermondiias as an effective biocontrol agent and alternative to chemical fungicides to control the blue mold of apple fruit.

Keywords: yeast, yeast enzymes, biocontrol, post harvest diseases

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Authors: V. Hakimzadeh, M. Noori, M. maleki

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The aim of this study was to determine the composition and antimicrobial effect of Mentha piperita essential oil in "in-vitro" condition. The chemical composition of the essential oil obtained by hydro-distillation was examined by GC/MS and the antimicrobial effect was studied on the growth of seven microbial species including Bacillus cereus, Pseudomonas aeruginosa and Proteus vulgaris using micro-dilution method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined. Chemical composition analysis identified a total of 28 compounds in which the main components were menthol (32%), mentone (13.4), menthyl acetate (12%), 1,8-cineole (8.2%) and neomenthol (4%) representing 69.6 % of the total oil. Other separated components accounted for less than 30.4% of the oil. Results of antimicrobial analysis showed that the MIC values for Bacillus cereus, Pseudomonas aeruginosa and Proteus vulgaris was respectively 50, 200 and 100 µg/ml and the MBC was determined at 200, 400 and 200 µg/ml respectively. The results of the present study indicated that Mentha piperita essential oil had significant antimicrobial activity.

Keywords: antimicrobial activity, essential oil composition, Mentha piperita

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Authors: Haowei Chen, Kaiqi Xiong

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This paper aims to answer how malware will propagate in Wireless Mesh Networks (WMNs) and how communication radius and distributed density of nodes affects the process of spreading. The above analysis is essential for devising network-wide strategies to counter malware. We answer these questions by developing an improved dynamical system that models malware propagation in the area where nodes were uniformly distributed. The proposed model captures both the spatial and temporal dynamics regarding the malware spreading process. Equilibrium and stability are also discussed based on the threshold of the system. If the threshold is less than one, the infected nodes disappear, and if the threshold is greater than one, the infected nodes asymptotically stabilize at the endemic equilibrium. Numerical simulations are investigated about communication radius and distributed density of nodes in WMNs, which allows us to draw various insights that can be used to guide security defense.

Keywords: Bluetooth security, malware propagation, wireless mesh networks, stability analysis

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Authors: Reginald Chikere Keke

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This paper examined leadership failure consequent on endemic corruption as being the bane of good governance in Nigeria since independence in 1960 and the way forward. Nigeria is lavishly gifted by nature of abundance in human and material resources to be harnessed a strategic, resolute, ingenious, and inventive leadership. For leadership to drive sustainable growth in society, it must be rooted in the cultural values of the people. This, however, is contrary in Nigeria owing to unscrupulous leadership miscarriage, corruption, and bad governance. Using the eclectic approach, the paper scrutinizes the issues of leadership, corruption, and governance to clearly show how bad leadership and governance have destroyed the national fabric and the way out of Nigeria's development quack mire. Furthermore, this paper examined the perplexing nature of corruption in Nigeria that has made it the only lucrative endeavor for politicians and their cronies, leading Nigeria to be regarded as the world's poverty capital. This paper advocate that Nigerians and the international community must endeavor to enshrine effective leadership and good governance through strong institutions, laws, and individuals who have zero tolerance for corruption and mediocrity in the polity. Only then will the fatherland of everyone’s dreams will be realized, and the labors of our hero’s past will not be in vain.

Keywords: corruption, leadership, governance, Nigeria

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Authors: Marie Bajnarová

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The core part of the research project is represented by taking a perspective on the role of an educator in Juvenile Institutional Centres. In accordance with the research questions, the research explores impact of the environment, situations, practices, attitudes, values and also experience of the respondents. Art activities minimize risky behaviours and contribute to a healthy lifestyle. They also help children and adolescents with conduct disorders develop positive social behaviour, psychosocial skills and cope with difficult life situations.

Keywords: Juvenile Detention Centres, drawing, conduct disorders, art therapy

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Authors: Abiodun Amusan, Olugbenga Akinola, Kazeem Akano, María Hernández-Castañeda, Jenna Dick, Akintunde Sowunmi, Geoffrey Hart, Grace Gbotosho

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Introduction: Artemisinin-based Combination Therapy (ACTs) is the cornerstone malaria treatment option in most malaria-endemic countries. Unfortunately, the malaria control effort is constantly being threatened by resistance of Plasmodium falciparum to ACTs. The recent evidence of artemisinin resistance in East Africa and its possibility of spreading to other African regions portends an imminent health catastrophe. This study aimed at evaluating the occurrence, prevalence, and influence of artemisinin-resistance markers on treatment outcomes in Ibadan before and after post-adoption of artemisinin combination therapy (ACTs) in Nigeria in 2005. Method: The study involved day zero dry blood spot (DBS) obtained from malaria patients during retrospective (2000-2005) and prospective (2021) studies. A cohort in the prospective study received oral dihydroartemisinin-piperaquine and underwent a 42-day follow-up to observe treatment outcomes. Genomic DNA was extracted from the DBS samples using a QIAamp blood extraction kit. Fragments of P. falciparum kelch13 (Pfkelch13), P. falciparum coronin (Pfcoronin), P. falciparum multidrug resistance 2 (PfMDR2), and P. falciparum chloroquine resistance transporter (PfCRT) genes were amplified and sequenced on a sanger sequencing platform to identify artemisinin resistance-associated mutations. Mutations were identified by aligning sequenced data with reference sequences obtained from the National Center for Biotechnology Information. Data were analyzed using descriptive statistics and student t-tests. Results: Mean parasite clearance time (PCT) and fever clearance time (FCT) were 2.1 ± 0.6 days (95% CI: 1.97-2.24) and 1.3 ± 0.7 days (95% CI: 1.1-1.6) respectively. Four mutations, K189T [34/53(64.2%)], R255K [2/53(3.8%)], K189N [1/53(1.9%)] and N217H [1/53(1.9%)] were identified within the N-terminal (Coiled-coil containing) domain of Pfkelch13. No artemisinin resistance-associated mutation usually found within the β-propeller domain of the Pfkelch13 gene was found in these analyzed samples. However, K189T and R255K mutations showed a significant correlation with longer parasite clearance time in the patients (P<0.002). The observed Pfkelch13 gene changes did not influence the baseline mean parasitemia (P = 0.44). P76S [17/100 (17%)] and V62M [1/100 (1%)] changes were identified in the Pfcoronin gene fragment without any influence on the parasitological parameters. No change was observed in the PfMDR2 gene, while no artemisinin resistance-associated mutation was found in the PfCRT gene. Furthermore, a sample each in the retrospective study contained the Pfkelch13 K189T and Pfcoronin P76S mutations. Conclusion: The study revealed absence of genetic-based evidence of artemisinin resistance in the study population at the time of study. The high frequency of K189T Pfkelch13 mutation and its correlation with increased parasite clearance time in this study may depict geographical variation of resistance mediators and imminent artemisinin resistance, respectively. The study also revealed an inherent potential of parasites to harbour drug-resistant genotypes before the introduction of ACTs in Nigeria.

Keywords: artemisinin resistance, plasmodium falciparum, Pfkelch13 mutations, Pfcoronin

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Authors: Getachew Tilahun, Oluwole Makinde, David Malonza

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We propose and analyze a non-linear mathematical model for the transmission dynamics of pneumonia disease in a population of varying size. The deterministic compartmental model is studied using stability theory of differential equations. The effective reproduction number is obtained and also the local and global asymptotically stability conditions for the disease free and as well as for the endemic equilibria are established. The model exhibit a backward bifurcation and the sensitivity indices of the basic reproduction number to the key parameters are determined. Using Pontryagin’s maximum principle, the optimal control problem is formulated with three control strategies; namely disease prevention through education, treatment and screening. The cost effectiveness analysis of the adopted control strategies revealed that the combination of prevention and treatment is the most cost effective intervention strategies to combat the pneumonia pandemic. Numerical simulation is performed and pertinent results are displayed graphically.

Keywords: cost effectiveness analysis, optimal control, pneumonia dynamics, stability analysis, numerical simulation

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Authors: Messaouda Belaid, Salima Kebbouche-Gana, Djamila Benaziza

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Our study focuses on determining the antibacterial activity of some honeys from northern Algeria. To test this activity, the agar well diffusion methods was employed. The bacterial strains tested were Staphylococcus aureus, Bacillus subtilis, Streptococcus faecalis, Klebsiella pneumoniae, Escherichia coli, and Pseudomonas aeroginosae. The results showed that all the microbes tested were inhibited by all honey used in this study but Those bacteria that appear to be more sensitive to all honey tested are Staphylococcus aureus and Pseudomonas aeroginosae.

Keywords: honey, antibacterial activity, Northern Algeria, Staphylococcus aureus

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Authors: Yunusa Aliyu Hadejia

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Cholera is a gastrointestinal disease caused by a bacterium called Vibrio Cholerae which spread as a result of eating food or drinking water contaminated with feaces from an infected person. In this work we proposed and analyzed the impact of isolating infected people and give them therapeutic treatment, the specific objectives of the research was to formulate a mathematical model of cholera transmission incorporating treatment class, to make analysis on stability of equilibrium points of the model, positivity and boundedness was shown to ensure that the model has a biological meaning, the basic reproduction number was derived by next generation matrix approach. The result of stability analysis show that the Disease free equilibrium was both locally and globally asymptotically stable when R_0< 1 while endemic equilibrium has locally asymptotically stable when R_0> 1. Sensitivity analysis was perform to determine the contribution of each parameter to the basic reproduction number. Numerical simulation was carried out to show the impact of the model parameters using MAT Lab Software.

Keywords: mathematical model, treatment, stability, sensitivity

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Authors: Nassibeh Hosseini-Vasoukolaei, Amir Ahmad Akhavan, Mahmood Jeddi-Tehrani, Ali Khamesipour, Mohammad Reza Yaghoobi Ershadi, Kamhawi Shaden, Valenzuela Jesus, Hossein Mirhendi, Mohammad Hossein Arandian

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Zoonotic cutaneous leishmaniasis (ZCL) is an endemic disease in many rural areas of Iran. Sand flies were collected from rural areas of Esfahan province and were identified using valid identification keys. DNA was extracted from sand flies and Nested PCRs were done using specific primers. In this study, 44 out of 152 (28.9 %) sand flies were infected with L. majoralone. Eight sand flies showed mixed infection: four sand flies (2.6 %) were infected with L. major, L. turanicaand L. gerbili, one sand fly (0.7 %) was infected with L. major and L. turanica and three sand flies (2 %) were infected with L. turanicaand L. gerbili. Our results demonstrate the natural infection of P. papatasi sand fly with three species of L. major, L. turanica and L. gerbili which are circulating among R. opimusreservoir host and P. papatasi sand fly vector in central Iran.

Keywords: Phlebotomus papatasi, Leishmania major, Leishmania turanica, Leishmania gerbili, mixed infection, Iran

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Authors: Abu S. Mustafa, Mohammad W. Khan, Faraz Shaheed Khan, Nazima Habibi

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Brucellosis is a zoonotic disease of worldwide prevalence. There are at least four species and several strains of Brucella that cause human disease. Brucella genomes have very limited variation across strains, which hinder strain identification using classical molecular techniques, including PCR and 16 S rDNA sequencing. The aim of this study was to perform whole genome sequencing of clinical isolates of Brucella and perform bioinformatics and comparative genomics analyses to determine the existence of genetic differences across the isolates of a single Brucella species and strain. The draft sequence data were generated from 15 clinical isolates of Brucella melitensis (biovar 2 strain 63/9) using MiSeq next generation sequencing platform. The generated reads were used for further assembly and analysis. All the analysis was performed using Bioinformatics work station (8 core i7 processor, 8GB RAM with Bio-Linux operating system). FastQC was used to determine the quality of reads and low quality reads were trimmed or eliminated using Fastx_trimmer. Assembly was done by using Velvet and ABySS softwares. The ordering of assembled contigs was performed by Mauve. An online server RAST was employed to annotate the contigs assembly. Annotated genomes were compared using Mauve and ACT tools. The QC score for DNA sequence data, generated by MiSeq, was higher than 30 for 80% of reads with more than 100x coverage, which suggested that data could be utilized for further analysis. However when analyzed by FastQC, quality of four reads was not good enough for creating a complete genome draft so remaining 11 samples were used for further analysis. The comparative genome analyses showed that despite sharing same gene sets, single nucleotide polymorphisms and insertions/deletions existed across different genomes, which provided a variable extent of diversity to these bacteria. In conclusion, the next generation sequencing, bioinformatics, and comparative genome analysis can be utilized to find variations (point mutations, insertions and deletions) across different genomes of Brucella within a single strain. This information could be useful in surveillance and epidemiological studies supported by Kuwait University Research Sector grants MI04/15 and SRUL02/13.

Keywords: brucella, bioinformatics, comparative genomics, whole genome sequencing

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Authors: O. P. Yadav

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Ag-N co-doped ZnS and ZnS/Fe2O3 composite nanoparticles have been synthesized by chemical and sol-gel methods. As-synthesized nanomaterial have been characterized by XRD and TEM techniques and their antimicrobial effects were studied using paper disc diffusion technique against gram positive (Staphylococcus aureus) and gram negative (Escherichia coli) bacteria. As-synthesized nanomaterial showed potent antimicrobial activity against studied bacterial strains. Antimicrobial activity of synthesized nanomaterial has also been compared with some commonly used antibiotics.

Keywords: antibiotic, Escherichia coli, nanomaterial, TEM, Staphylococcus aureus

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Authors: R. J. Kuo, W. C. Cheng, W. C. Lien, T. J. Yang

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Taiwan is a hyper endemic area for the Hepatitis B virus (HBV). The estimated total number of HBsAg carriers in the general population who are more than 20 years old is more than 3 million. Therefore, a case record review is conducted from January 2003 to June 2007 for all patients with a diagnosis of acute hepatitis who were admitted to the Emergency Department (ED) of a well-known teaching hospital. The cost for the use of medical resources is defined as the total medical fee. In this study, principal component analysis (PCA) is firstly employed to reduce the number of dimensions. Support vector regression (SVR) and artificial neural network (ANN) are then used to develop the forecasting model. A total of 117 patients meet the inclusion criteria. 61% patients involved in this study are hepatitis B related. The computational result shows that the proposed PCA-SVR model has superior performance than other compared algorithms. In conclusion, the Child-Pugh score and echogram can both be used to predict the cost of medical resources for patients with acute hepatitis in the ED.

Keywords: acute hepatitis, medical resource cost, artificial neural network, support vector regression

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Authors: Sze Wing Ho, Nagendra P. Shah

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Lactic acid bacteria (LAB) have shown the beneficial effects on human gastrointestinal tract, such as protects diarrhea induced by lactose intolerance or enteric pathogens. Citrulline is a non-protein amino acid and also the precursors of arginine and nitric oxide, it has shown to enhance intestinal barrier function. Citrulline has shown to improve the growth of some strains of LAB, it is important for LAB to have a sufficient cell concentration to contribute the effects. Therefore, the aims of this study were to investigate the effect of combining citrulline with LAB on the anti-adhesion effect against pathogens and the effect on the cell integrity. The effect of citrulline on selected LAB was determined by incubating in 0%, 0.1% or 0.2% citrulline enriched MRS broth for 18 h. The adhesion ability of LAB and the anti-adhesion effect of LAB and citrulline against pathogens were performed on IPEC-J2 cell line. Transepithelial electrical resistance (TEER) assay was used to measure the tight junction (TJ) integrity. TJ proteins (claudin-1, occludin and zonula occluden-1 (ZO-1)) were determined by western blot analysis. It found that the growth of Lactobacillus helveticus ASCC 511 was significantly stimulated by 0.2% citrulline compared with control during 18 h fermentation. The adhesion of L. helveticus ASCC 511 and Lactobacillus delbrueckii ssp. bulgaricus (L. bulgaricus) ASCC 756 was increased when supplemented with citrulline. Citrulline has shown significant inhibitory effect on the adhesion of Escherichia coli PELI0480 (O157:H7), Shigella sonnei ATCC 25931, Staphyloccocus aureus CMCC26003 and Cronobacter sakazakii ATCC 29544. The anti-adhesion effect of L. helveticus ASCC 511, L. bulgaricus ASCC 756 and Lactobacillus paracasei ASCC 276 against Cronobacter sakazakii ATCC 29544 was significantly enhanced with citrulline supplementation. Treatments with citrulline and LAB were able to maintain the TEER of IPEC-J2 cell and shown the positive effect on the TJ proteins. In conclusion, citrulline had stimulating effect on some strains of LAB and determined to improve the adhesion of LAB on intestinal epithelial cell, to enhance the inhibitory effect on enteric pathogens adhesion as well as had beneficial effects on maintaining cell integrity. It implied LAB supplemented with citrulline might have advantageous effects on gastrointestinal tracts.

Keywords: citrulline, lactic acid bacteria, amino acid, anti-adhesion effect, cell integrity

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Authors: S. K. Hemalata, M. N. Shiva Kameshwari

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The present work focusses on anti-inflammatory action of Urginea wightii in-vitro. Urginea wightii is a member of Hyacinthaceae and considered to be wonder plant because of its varied important medicinal properties. The plant is endemic to India, Africa, and Mediterranian regions. Presence of alkaloids, flavonoid-glycosides especially flavonone derivatives are responsible for the strong anti-inflammatory activity of Urginea wightii. In present research work, anti-inflammatory activity of methanol extract of the bulb powder was tested on Male Wistar Rats. In these test animals, inflammation was induced by injecting carrageenan as the irritant to induce paw edema in Wistar rats. Inflammation of Paw edema was treated with both plant extract and Pyrox gel a known synthetic anti-inflammatory drug through external application. The result indicated that anti-inflammatory activity of Urginea wightii extract was almost similar to the synthetic Pyrox gel. This disproves the modern world's scepticism towards the herbal medicines and encourages to rely on natural plant extracts.

Keywords: anti-inflammatory activity, flavonoid-glycosides, Pyrox gel, Urginia wightii

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Authors: Ellana Boada, Eric Santos-Clotas, Alba Cabrera-Codony, Maria Martin, Lluis Baneras, Frederic Gich

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Volatile methylsiloxanes (VMS) are a group of manmade silicone compounds widely used in household and industrial applications that end up on the biogas produced through the anaerobic digestion of organic matter in landfills and wastewater treatment plants. The presence of VMS during the biogas energy conversion can cause damage on the engines, reducing the efficiency of this renewable energy source. Non regenerative adsorption onto activated carbon is the most widely used technology to remove siloxanes from biogas, while new trends point out that biotechnology offers a low-cost and environmentally friendly alternative to conventional technologies. The first objective of this research was to enrich, isolate and identify bacterial species able to grow using siloxane molecules as a sole carbon source: anoxic wastewater sludge was used as initial inoculum in liquid anoxic enrichments, adding D4 (as representative siloxane compound) previously adsorbed on activated carbon. After several months of acclimatization, liquid enrichments were plated onto solid media containing D4 and thirty-four bacterial isolates were obtained. 16S rRNA gene sequencing allowed the identification of strains belonging to the following species: Ciceribacter lividus, Alicycliphilus denitrificans, Pseudomonas aeruginosa and Pseudomonas citronellolis which are described to be capable to degrade toxic volatile organic compounds. Kinetic assays with 8 representative strains revealed higher cell growth in the presence of D4 compared to the control. Our second objective was to characterize the community composition and diversity of the microbial community present in the enrichments and to elucidate whether the isolated strains were representative members of the community or not. DNA samples were extracted, the 16S rRNA gene was amplified (515F & 806R primer pair), and the microbiome analyzed from sequences obtained with a MiSeq PE250 platform. Results showed that the retrieved isolates only represented a minor fraction of the microorganisms present in the enrichment samples, which were represented by Alpha, Beta, and Gamma proteobacteria as dominant groups in the category class thus suggesting that other microbial species and/or consortia may be important for D4 biodegradation. These results highlight the need of additional protocols for the isolation of relevant D4 degraders. Currently, we are developing molecular tools targeting key genes involved in siloxane biodegradation to identify and quantify the capacity of the isolates to metabolize D4 in batch cultures supplied with a synthetic gas stream of air containing 60 mg m⁻³ of D4 together with other volatile organic compounds found in the biogas mixture (i.e. toluene, hexane and limonene). The isolates were used as inoculum in a biotrickling filter containing lava rocks and activated carbon to assess their capacity for siloxane removal. Preliminary results of biotrickling filter performance showed 35% of siloxane biodegradation in a contact time of 14 minutes, denoting that biological siloxane removal is a promising technology for biogas upgrading.

Keywords: bacterial cultivation, biogas upgrading, microbiome, siloxanes

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Authors: Soumeya Krimat, Tahar Dob, Mohamed Toumi, Aicha Kesouri, Hafidha Metidji, Chelghoum Chabane

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Lavandula antineae Maire is an endemic medicinal plant of Algeria which is traditionally used for the treatment of chills, bruises, oedema and rheumatism. The present study was designed to investigate the phytochemical screening, total phenolic and antioxidant activity of Lavandula antineae Maire for the first time. Phytochemical screening revealed the presence of different kind of chemical groups (anthraquinones, terpenes, saponins, flavonoids, tannins, O-heterosides, C-heterosides, phenolic acids). The amounts of total phenolics in the extracts (hydromethanolic and ethyl acetate extract) were determined spectrometrically. From the analyses, ethyl acetate extract had the highest total phenolic content (262.35 mg GA/g extract) and antioxidant activity (IC50=7.10 µg/ml) using DPPH method. The ethyl acetate extract was also more potent on reducing power compared to hydromethanolic extract. The results suggested that L. antineae could be considered as a new potential source of natural antioxidant for pharmaceuticals and food preservation.

Keywords: Lavandula antineae, antioxidant activity, phytochemical screening, total phenolics

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Authors: A. M. U. P. Kumari, Vidanapathirana. J., Amarasekara J., Karunanayaka L.

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Leptospirosis is a zoonotic infection with significant morbidity and mortality. As an occupational disease, it has become a global concern due to its disease burden in endemic countries and rural areas. The aim of this study was to assess disease burden in terms of DALYs of leptospirosis. A hospital-based descriptive cross-sectional study was conducted using 450 clinically diagnosed leptospirosis patients admitted to base and above hospitals in Monaragala district, Sri Lanka, using a pretested interviewer administered questionnaire. The patients were followed up till normal day today life after discharge. Estimation of DALYs was done using laboratory confirmed leptospirosis patients. Leptospirosis disease burden in the Monaragala district was 44.9 DALYs per 100,000 population which includes 33.18 YLLs and 10.9 YLDs. The incidence of leptospirosis in the Monaragala district during the study period was 59.8 per 100,000 population, and the case fatality rate (CFR) was 1.5% due to delay in health seeking behaviour; 75% of deaths were among males due to multi organ failure. The disease burden of leptospirosis in the Moneragala district was significantly high, and urgent efforts to control and prevent leptospirosis should be a priority.

Keywords: human leptospirosis, disease burden, disability adjusted life Years, Sri Lanka

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Authors: Waheed Ahmad Safi, Shunichi Nakamura

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Concrete filled steel I-girder (CFIG) bridge was proposed and the bending and shear strength was confirmed by experiments. The area surrounded by the upper and lower flanges and the web is filled with concrete in CFIG, which is used to the intermediate support of a continuous girder. Three-dimensional finite element models were established to simulate the bending and shear behaviors of CFIG and to clarify the load transfer mechanism. Steel plates and filled concrete were modeled as a three-dimensional 8-node solid element and steel reinforcement bars as a three-dimensional 2-node truss element. The elements were mostly divided into the 50 x 50 mm mesh size. The non-linear stress-strain relation is assumed for concrete in compression including the softening effect after the peak, and the stress increases linearly for concrete in tension until concrete cracking but then decreases due to tension stiffening effect. The stress-strain relation for steel plates was tri-linear and that for reinforcements was bi-linear. The concrete and the steel plates were rigidly connected. The developed FEM model was applied to simulate and analysis the bending behaviors of the CFIG specimens. The vertical displacements and the strains of steel plates and the filled concrete obtained by FEM agreed very well with the test results until the yield load. The specimens collapsed when the upper flange buckled or the concrete spalled off. These phenomena cannot be properly analyzed by FEM, which produces a small discrepancy at the ultimate states. The FEM model was also applied to simulate and analysis the shear tests of the CFIG specimens. The vertical displacements and strains of steel and concrete calculated by FEM model agreed well with the test results. A truss action was confirmed by the FEM and the experiment, clarifying that shear forces were mainly resisted by the tension strut of the steel plate and the compression strut of the filled concrete acting in the diagonal direction. A trail design with the CFIG was carried out for a four-span continuous highway bridge and the design method was established. Construction cost was estimated about 12% lower than that of a conventional steel I-section girder.

Keywords: concrete filled steel I-girder, bending strength, FEM, limit states design, steel I-girder, shear strength

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Authors: Fang-Bo Yu, Li-Bo Guan, Sheng-Dao Shan

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Biogas slurry was mixed with six commercial fungicides and screening against 11 phytopathogens was carried out. Results showed that inhibition of biogas slurry was different for the test strains and no significant difference between treatments of Didymella bryoniae, Fusarium oxysporum f. sp. vasinfectum, Aspergillus niger, Rhizoctonia cerealis, F. graminearum and Septoria tritici was observed. However, significant differences were found among Penicillium sp., Botrytis cinerea, Alternaria sonali, F. oxysporum F. sp. melonis and Sclerotinia sclerotiorum. The approach described here presents a promising alternative to current manipulation although some issues still need further examination. This study could contribute to the development of sustainable agriculture and better utilization of biogas slurry.

Keywords: anaerobic digestion, biogas slurry, phytopathogen, sustainable agriculture

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Authors: Zahia Kabouche, Ouissem Gherboudj, Naima Boutaghane, Ahmed Kabouche, Laurence Voutquenne-Nazabadioko

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Chrysanthemum herbs (Asteraceae) are extensively used as food additives and in folk medicine. Anti-cancer, anti-human immunodeficiency virus type 1 (HIV-1), anti-inflammatory, antinociceptive and antiproliferative activities as well as antioxidant effects have been reported for Chrysanthemum species. We report the isolation and identification of flavonoids and new and known terpenoids from the endemic species, C. macrocarpum and C. deserticolum “guertoufa”, used in Algerian Sahara as tea drinks and in “couscous” and soups “Chorba”. Structures of the isolated compounds were established by 1-D and 2-D homo and hetero-nuclear NMR (1H, 13C, COSY, HSQC, HMBC, and NOESY), mass spectrometry, UV and comparison with literature data. C. deserticolum extracts were tested by four methods to identify the antioxidant activity namely, ABTS•+, DPPH• scavenging, CUPRAC and ferrous-ions chelating activity methods. Anti-inflammatory, antinociceptive, antiproliferative and antioxidant activities of C. macrocarpum extracts and isolated compounds are also reported here.

Keywords: Chrysanthemum macrocarpum, C. deserticolum, flavonoids, terpenoids, antioxidant, anti-inflammatory, anti-proliferative

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Authors: Omaima A. Sharaf, Tarek A. Moussa, Said M. Badr El-Din, H. Moawad

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Background: Polycyclic aromatic hydrocarbons (PAHs) pose great environmental and human health concerns for their widespread occurrence, persistence, and carcinogenic properties. PAHs releases due to anthropogenic activities to the wider environment have led to higher concentrations of these contaminants than would be expected from natural processes alone. This may result in a wide range of environmental problems that can accumulate in agricultural ecosystems, which threatened to become a negative impact on sustainable agricultural development. Thus, this study aimed to evaluate the physico-chemical, and microbial properties of the compost leachate (CL) to assess its role as nutrient and microbial source (biostimulation/bioaugmentation) for developing a cost-effective bioremediation technology for PAHs contaminated sites. Material and Methods: PAHs-degrading bacteria were isolated from CL that was collected from a composting site located in central Scotland, UK. Isolation was carried out by enrichment using phenanthrene (PHR), pyrene (PYR) and benzo(a)pyrene (BaP) as the sole source of carbon and energy. The isolates were characterized using a variety of phenotypic and molecular properties. Six different isolates were identified based on the difference in morphological and biochemical tests. The efficiency of these isolates in PAHs utilization was assessed. Further analysis was performed to define taxonomical status and phylogenic relation between the most potent PAHs-utilizing bacterial strains and other standard strains, using molecular approach by partial 16S rDNA gene sequence analysis. Results indicated that the 16S rDNA sequence analysis confirmed the results of biochemical identification, as both of biochemical and molecular identification of the isolates assigned them to Bacillus licheniformis, Pseudomonas aeruginosa, Alcaligenes faecalis, Serratia marcescens, Enterobacter cloacae and Providenicia which were identified as the prominent PAHs-utilizers isolated from CL. Conclusion: This study indicates that the CL samples contain a diverse population of PAHs-degrading bacteria and the use of CL may have a potential for bioremediation of PAHs contaminated sites.

Keywords: polycyclic aromatic hydrocarbons, physico-chemical analyses, compost leachate, microbial and biochemical analyses, phylogenic relations, 16S rDNA sequence analysis

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Authors: Omaima A. Sharaf, Wafaa M. Abd El-Rahim, Hassan Moawad, Michael J. Sadowsky

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Textile industry is one of the important industries in the worldwide. It is known that the eco-friendly industrial and agricultural activities are significant for socio-economic stability of all countries. The absence of appropriate industrial waste water treatments is essential barrier for sustainable development in food and agricultural sectors especially in developing country like Egypt. Thus, the development of enzymatic bioremediation technology for textile dye removal will enhance the collaboration between scientists who develop the technology and industry where this technology will be implemented towards the safe disposal of the textile dye wastes. Highly efficient microorganisms are of most importance in developing and using highly effective biological treatment processes. Bacterial degradation of azo dyes is generally initiated by an enzymatic step that involves cleavage of azo linkages, usually with the aid of an azoreductase as electron donor. Thus, expanding the spectrum of microorganisms with high enzymatic activities as azoreductases and discovering novel azo-dye degrading enzymes, with enhanced stability and superior catalytic properties, are necessary for many environmental and industrial applications. Consequently, the use of molecular tools has become increasingly integrated into the understanding of enzyme properties and characterization. Researchers have utilized a gene cloning and expression methods as a tool to produce recombinant protein for decolorizing dyes more efficiently. Thus, presumptive evidence for the presence of genes encoding azoreductases in the genomes of selected local, and most potent azo-degrading strains were obtained by using specific oligonucleotides primers. These potent strains have been isolated from textile industrial wastewater in Egypt and identified using 16S rRNA sequence analysis as 'Lysinibacillus macrolidesB8, Brevibacillus parabrevisB11, Bacillus coagulansB7, and B. cereusB5'. PCR products of two full length genes designated as (AZO1;621bp and AZO2;534bp) were detected. BLASTx results indicated that AZO1 gene was corresponding to predicted azoreductase from of Bacillus sp. ABP14, complete genome, multispecies azoreductase [Bacillus], It was submitted to the gene bank by an accession no., BankIt2085371 AZO1 MG923210 (621bp; 207 amino acids). AZO1 was generated from the DNA of our identified strains Lysinibacillus macrolidesB8. On the other hand, AZO2 gene was corresponding to a predicted azoreductase from Bacillus cereus strain S2-8. Gene bank accession no. was BankIt2085839 AZO2 MG932081 (534bp;178 amino acids) and it was amplified from our Bacillus coagulansB7. Both genes were successfully cloned into pCR2.1TOPO (Invitrogen) and in pET28b+ vectors, then they transformed into E. coli DH5α and BL21(DE3) cells for heterologous expression studies. Our recombinant azoreductases (AZO1&AZO2) exhibited potential enzyme activity and efficiently decolorized an azo dye (Direct violet). They exhibited pH stability between 6 and 8 with optimum temperature up to 60°C and 37 °C after induction by 1mM and 1.5mM IPTG, for both AZO1 &AZO2, respectively. These results suggested that further optimization and purification of these recombinant proteins by using different heterologous expression systems will give great potential for the sustainable utilization of these recombinant enzymes in several industrial applications especially in wastewater treatments.

Keywords: azoreductases, decolorization, enzyme activity, gene cloning and expression

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Authors: Diobein Flores, Venus Buagas, Virgie Darunday

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The study inventoried the species composition of terrestrial ferns and lycopods in Center for Ecological Development and Recreation (CEDAR) Impalutao, Impasug-ong, Bukidnon. Specifically, it aimed to determine and describe the species composition, and diagnostic characters of the ferns and lycopods in the study site. Transect walk method was employed in the inventory of the species. Each species were classified, identified and described according to its diagnostic characters. Results of the study revealed a total of 20 species of ferns and lycopods. Of these, 18 species were ferns and 2 species were lycopods. Eleven (11) families and fifteen (15) genera for ferns and one (1) family and one (1) genera for lycopods. Psomiocarpa apifolia is Philippine endemic and said to be vulnerable or threatened. Taxonomic characters based on habit, rhizome, leaf arrangement and orientation, stem structure and circinate vernation were used to identify the terrestrial pteridophtyes into families, genera and species. The species collected and assessment in CEDAR should be further investigated and monitor their conservation status.

Keywords: alpha taxonomy, conservation, habit, taxonomic characters

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Authors: Fouzia Qamar, Shahida Hasnain

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The present study is primarily concerned with the alteration in haemocyte profile of adult male Periplaneta americana with emphasis on the effect of bacterial inoculations on the haemogram i.e., total haemocyte count (THC) and differential haemocyte count (DHC) of different haemocyte types of the target insect. Haemolymph cellular profile showed considerable alterations under the effect of nine strains of Agrobacterium tumefaciens after 8, 16 and 24 hrs of treatment thereby signifying the potential role of Agrobacterium tumefaciens as a possible biocontrol agent against the house hold pests.

Keywords: Agrobacterium tumefaciens, Periplaneta americana, Haemolymph, cellular parametes

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