Bacillus spp with an Antibacterial Activity | Chapter 13 | Recent Advances in Biological Research Vol. 2

Bacterial resistance to antibiotics poses a serious challenge to chemotherapy. Indeed, in recent decades, the risk of contracting infectious diseases has increased significantly due to the emergence and spread of multi-drug-resistant bacteria. The whole world is thus confronted with this situation which motivated scientists to find out new active molecules with an antibacterial effect. Therefore, the present investigation aims to isolate and identify microorganisms secreting substances with an antibacterial activity and to partially characterize their extracts. Thus, six bacteria were isolated from soil and water of different biotopes of Fez Morocco having an antibacterial effect against M. smegmatis, M. aurum, Gram-positive and Gram-negative bacteria. Based on biochemical characterization and 16S rDNA sequence analysis, it was revealed that the isolates belong to the genus Bacillus. The antibacterial activity of three of them was fully affected by proteases (pepsin and proteinase K) and heat treatment at 80ºC and 100ºC but it was stable at 4ºC for a month and 37ºC during 3h. The lost of the activity suggests a proteinaceous nature of the bio-active compounds, which might be useful in the development of antibacterial agents after their total purification in further work against bacterial infections.

Biography of author(s)

Dr. Ilham Zahir
Laboratory of Microbial Biotechnology, Department of Biology, Faculty of Sciences and Technical, University Sidi Mohamed Ben Abdellah, BP 2202, Road of Immouzer, Fez, Morocco and Polyvalent Laboratory in Research and Development, Department of Biology, Polydisciplinary Faculty, Sultan Moulay Slimane University, Beni Mellal, Morocco.

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Evaluation of Three Phenotypic Tests vs. Duplex (coa & mecA) PCR for Detection of Methicillin-Resistance Staphylococcus aureus (MRSA) and S. epidermidis (MRSE) Strains from Patients with Nosocomial Infections (NI) in Several Mexican Hospitals | Chapter 12 | Recent Advances in Biological Research Vol. 2

Background: Staphylococcus aureus (SA) and Staphylococcus epidermidis are a leading cause of hospital acquired infections worldwide. Epidemiological features have also changed in Mexican hospitals and either methicillin-resistance Staphylococcus aureus (MRSA) or Staphylococcus epidermidis (SERM) infections appear to be an emerging phenomenon.  

Aims: The first aim of the present study was to detect the amplification of both pairs, coa and mecA genes by duplex PCR in MRSA from Mexican patients with NI and the comparison of these results versus three routinely phenotypic MRSA-detection methods.The second aim of this study was to apply duplex PCR to exclusively detect the same pair of genes for MRSA or MRSE strains from patients with NI in several national hospitals.

Place and Duration of Study: Department of Public Health, Faculty of Medicine, National Autonomous University of Mexico (UNAM) in collaboration with six General Hospitals belonging to the Health Care & Social Services Institute for Workers (ISSSTE), the first one HGC (Hospital Gonzálo Castañeda Escobar), during a year span of August of 2004 to December of 2005, the second study was performed through an extension of a collaborative study of the five hospitals (CMN20N; FQG; LALM; GIZ & 1°Oct), during a year period of May of 2012 through November of 2013 and the seventh hospital from the third collaboration study through the National Institute of Perinatology (INPer), during a time period of August of 2012 through December of 2013.

Methodology: Among SA from HGC (n=100) strains isolated from patients with NI, 40% were MRSA. All SA strains were tested by various methods: cefoxitin (Cfx) disk-diffusion assay (DD), automated MicroScan System (MS), Oxacillin (Ox) (range 0.5 – 4.0 µg/ml) minimum inhibitory concentration (MIC), PBP2a latex agglutination test (LA) (Oxoid) and the molecular amplification by PCR of the coa and mecA genes. SA strains were previously identified by MS, and both the tube coagulase (TCT) and mannitol (MT) tests. From the second and third collaborative study, a total of 345 staphylococci strains were isolated from ISSSTE´s hospitals and INPer´s hospitals,129 SA strains from ISSSTE and 246 SE strains from INPer. These clinical (SA or SE) strains were first phenotypical characterized in both health institutions and immediately after were molecularly characterized by Duplex PCR detection of species-specific and antibiotic-resistance genes, in the Public Health Laboratory at UNAM.

Results: Out of 100 SA strains from HGC, 35 were confirmed as MRSA isolates by detection of mecA gene by PCR in addition to only 5 strains verified phenotypically but coa gen negative. These 35 MRSA strains and 53 mecA gen negative isolates, as well as 5 strains phenotypically characterized (MSSA) from NI, were assayed with all 3 laboratory tests. Only 2 clinical strains were negative to both genes: coa & mecA, but positive to all phenotypic tests. Using mecA as a gold standard (GS), category agreement for the 3 tests was: A method with higher sensitivity (SN) and specificity (SP) (SN/SP) was Cfx-DD (100/88) than other 2 methods Ox-MS (90/73) and LA (63/90). Using coa gene as a GS, category agreement for MS, and both TCT & MT were: 93/100, 95/98, and 91/98, respectively. From the second and third collaborative study, out of 129 SA strains from ISSSTE and INPer, 93% were detected by PCR as coa+, therefore 93% (120/129) of these strains were confirmed as SA. Out of 120 SA strains, 88% (106/120) were confirmed as mecA+ by PCR detection, therefore these strains are MRSA. Out of 41 SA strains from INPer, 90% (37/41) were confirmed as SA and from these strains only 43% (16/37) were confirmed positive for mecA+ therefore these strains are MRSA. As a result of third collaboration study (INPer), out of 226 SE strains, only 90% (203/226) were confirmed by PCR as coa- and 85% (172/202) of these strains were detected by PCR as mecA+, therefore these strains are MRSE.

Conclusions: Molecular detection by PCR-duplex of specie specific coa genes and antibiotic resistance mecA genes by duplex PCR in MRSA and MRSE strains, is a very useful and powerful method to rapid and precisely discriminate typical MRSA and MRSE strains in the laboratory. A Cfx-DD test performed much better for detection of MRSA strains than those using OX-MS and LA. Both assays, Cfx-DD and PCR-duplex, represent a simple, rapid, reliable approaches for the detection of methicillin resistant staphylococci and could be applied to all national hospitals, to implement fast and adequate anti-MRSA and anti-MRSE therapy. Higher percentages of MRSA strains (88% vs. 43%) were isolated from ISSSTE´s hospitals than in INPer´s hospitals, could be due to the age of patients studied, mainly neonates from INPer than older patients from ISSSTE´s hospitals.

Biography of author(s)

Rubén Morelos-Ramírez
Department of Public Health, Faculty of Medicine, Universidad Nacional Autónoma de México (UNAM), Av. Universidad #3000, Coyoacán, 4510, México D. F, México

Roberto Cabrera-Contreras
Department of Public Health, Faculty of Medicine, Universidad Nacional Autónoma de México (UNAM), Av. Universidad #3000, Coyoacán, 4510, México D. F, México.

Enrique Meléndez-Herrada
Department of Public Health, Faculty of Medicine, Universidad Nacional Autónoma de México (UNAM), Av. Universidad #3000, Coyoacán, 4510, México D. F, México.

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Chemical Composition of Myrtus communis L. (Myrtaceae) Fruits | Chapter 11 | Recent Advances in Biological Research Vol. 2

The chemical composition of Myrtus communis L. extracts were prepared and analyzed by GC-MS. Sixteen phytochemical constituents of chemical compounds were identified in fruits of Myrtus communis. The relative percentage of linoleic acid methyl ester was high (27.19%), followed by oleic acid methyl ester (21.18%) and then octane 3,5- dimethyl (16.47%), dodecane (11.39%), palmitic acid methyl ester (6.80%) and tetradecane (6.69%) as well as, some components present in lower percentage such as stearic acid methyl ester (3.32%). GC-MS analysis revealed that 16 different chemical components were identified in the Myrtus communis, with high amount of linoleic acid (27.19%) helpful in revealing the pharmaceutical value of the plant. It also provides information about its trado-medical use. GC-MS analysis is important step towards understanding the nature of active ingredients in this medicinal plant and mentioned phytocompounds would be useful in the preparation of novel drugs for treating diseases.

Biography of author(s)

Karzan Omer Qader
Sulaimani University, College of Science, Department of Biology, Iraq.

Sahar A.A. Malik Al-Saadi
Basrah University, College of Science, Department of Biology, Iraq.

Thuraya Abdul Abbas Malik Al-Saadi
Al-Mustansiriya University, College of Basic Education, Department of Science, Iraq.

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Copper Nanomaterials as Delivery System in Combating Infective Agents and Cancer | Chapter 10 | Recent Advances in Biological Research Vol. 2

Infectious diseases portend serious threat hazards to public health globally due to the development of the resistance to broad spectrum antimicrobials, the emergence of multidrug resistant biofilms, the drug side-toxicity, the non-specificity of drug targeting and the inability to overcome biological barriers. In this context, nanotechnology-based metal nanoparticles have attracted attention as nanomedicine against different diseases for their additional capability to anchore other therapeutic contents effectively for delivery. Owing to the large surface area to volume ratio, copper nanomaterials have been utilized as potential anti-infective and anticarcinogenic agent in biomedical applications. Though their higher significant efficiencies in damaging pathogenic cells have been elucidated, their nano-metallic toxicity increases other side-toxicity to healthy cells. Therefore, copper, copper oxide and copper sulfide -nanoparticles should be surface functionalized with ligands and vesicular system to reduce the toxic side effect for delivering to specific site of interest in a sustained-release manner. The subject has been focused mainly on the synthesis, size and surface characteristics, mechanism of action and biomedical applications of copper nanomaterials in damaging various infective agents and cancer cells as probable potent drug delivery system.

Biography of author(s)

Ardhendu Kumar Mandal
Central Instrumentation Division, CSIR-Indian Institute of Chemical Biology, 4, Raja S.C. Mullick Road, Jadavpur, Kolkata – 700032, India

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Keratin Degradation by Penicillium purpurogenum and Aspergillus niger Isolated from Nigerian Soils Polluted with Tannery Wastes | Chapter 09 | Recent Advances in Biological Research Vol. 2

Background: Keratinophilic fungi are a group of fungi that colonize various keratinous substrates and degrade them to components of low molecular weight. Keratinized tissues include feathers, beaks of birds, horns, hooves, skins, hair, fur, wool, nails and claws of animals. Tannery soils are very rich in keratinous materials such as fur, wool and hair that are by-products of the tanning industry where hides and skins are processed. Keratinase producing fungi, Aspergillus niger and Penicillium purpurogenum were isolated from soils containing tannery wastes in Jos, Plateau State Nigeria.

Materials and Methods: Skim milk casein agar was employed to assay for extra cellular protease as an indication for keratinase production. The species had maximum proteolytic and keratinase activities in a Submerged Fermentation (SmF) using liquid basal medium supplemented with skim milk, chicken feathers and human hair as carbon and nitrogen sources. Effects of incubation temperature on protease and keratinase enzyme activity were also determined.

Results: The Spectrophotometric assay of the proteolytic and keratinolytic activity showed that P. purpurogenum had the highest activity (13.5 U/ml) on chicken feathers. This was followed by human hair with activity of 12 U/ml. The least activity of 11.9 U/ml was observed in skim milk medium. Similar results were recorded for A. niger with highest activity of 11.8 U/ml on chicken feathers. This was followed by human hair with activity of 10.6 U/ml while skim milk agar had the least activity of 10.0 U/ml. It was observed that incubation temperature had effects on the enzyme activity, with an optimum temperature of 37°C for both protease and keratinase.

Conclusion: These non-dermatophytic keratinolytic fungi may have potential use in biotechnological processes involving keratin hydrolysis. The results of this work reiterated that keratinolytic activity is relatively widespread among common fungi and may have an important role in keratin degradation in the natural environment. P. purpurogenum and A. niger bio-degraded skim milk casein, human hair and chicken feather in Submerged Fermentation (SmF). They had the greatest activity on chicken feathers indicating that they could be employed in waste and environmental pollution management. In Nigeria, poultry feathers, animal hair and other keratin sources including tannery wastes do not find suitable applications. Practical use of keratinase producing microorganisms such as the ones used in this study is being explored in applied microbiology where there is great need for active degraders for the management of keratin containing wastes.

Biography of author(s)

Dr. (Mrs.) Abigail Ify Ogbonna (Ph. D.)
Department of Plant Science and Biotechnology, University of Jos, P.M.B.2084 Jos, Nigeria

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Antibiotics Susceptibility Pattern of Methicillin Resistant Staphylococcus aureus (MRSA) in Enugu State, South-East Region of Nigeria | Chapter 08 | Recent Advances in Biological Research Vol. 2

Development of antimicrobial resistance by bacteria is now a worldwide health issue, as infection is one of the leading causes of death in the world today. The aim of this study was to evaluate the prevalence and antimicrobials susceptibility pattern of Methicilin-Resistant Staphylococcus aureus in 3 different hospitals in South-East geopolitical region of Nigeria. The identification and confirmation of the S. aureus were done using selective and differential medium (Mannitol salt agar) for S. aureus and by coagulase/staphylase test using Oxoid® reagents kits (DR0595A). The method used for antibiotics susceptibility pattern of the characterised S. aureus isolates was discs diffusion method, as recommended by the Clinical Laboratory Standards Institute (CLSI), discs containing oxacillin (5 µg/disc), vancomycin (30 µg/disc), cephalexin (30 µg/disc), levofloxacin (5 µg/disc), ciprofloxacin (5 µg/disc), tetracycline (30 µg/disc), cotrimoxazole (25 µg/disc), gentamicin (30 µg/disc), clindamycin (2 µg/disc) and rifampicin (5 µg/disc). MRSA confirmation was done using Oxoid® DR0900 penicillin binding protein (pbp2’) latex agglutination test kits. The results showed that out of 218 characterized clinical isolates, 39 of it were confirmed MRSA with varying percentages of resistance to various antibiotics thus: oxacillin (62.07%), vancomycin (60.35%), cephalexin (55.18%), levofloxacin (56.90%), ciprofloxacin (65.52%), tetracycline (68.97%), cotrimoxazole (67.25%), gentamicin (62.07%), clindamycin (63.79%) and rifampicin (62.07%). The S. aureus are more sensitive to Levofloxacin and less sensitive to tetracycline, clindamycin and rimfapicin. Latex agglutination test confirmed 39 strains of the clinical isolates to be MRSA. The results shows open wound as a source with highest prevalence and sputum with lowest prevalence of the MRSA with no significant change (P > 0.05).

Biography of author(s)

A. A. Agboke
Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmacy, University of Uyo, Uyo, Akwa Ibom State, Nigeria.

A. A. Attama
Departments of Pharmaceutics and Pharmaceutical Microbiology, Faculty of Pharmaceutical Sciences, University of Nigeria, Enugu State, Nigeria.

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Synthetic Elicitor-Induced Defense Responses in Tomato (Solanum lycopersicum) Cultivated in Côte d’Ivoire against Bacterial Wilt Caused by Ralstonia solanacearum | Chapter 07 | Recent Advances in Biological Research Vol. 2

Tomato is the most consumed vegetable in Côte d’Ivoire because it is used to cook most sauces and all forms of salad. Tomatoes annual yield fluctuates between 22,000 and 35,000 tons. The yield remains below demand. Several factors can explain these low yields. Among them, biotic factors are the most important. Among bacterial diseases impacting negatively tomato yield, bacterial wilt caused by Ralstonia solanacearum is a dangerous one that causes serious damage in the nursery as well as in plantation. It can cause up to 100% yield loss depending on the variety. Bacterial wilt caused by Ralstonia solanacearum still constitutes tomato (Solanum lycopersicum) economical disease in the world. This disease is favored by very humid soils and high temperatures. Genetic control currently used is the most effective strategy. Disease control in crops has also become possible through the induction of plant defense reaction. This work aims at assessing the effect of both synthetic elicitors (BABA and ASM) supposed to induce tomato defense reaction against bacteria and particularly Ralstonia solanacearum. BABA and ASM solutions were each applied at concentrations of 5 to 100 ppm on R. solanacearum in vitro, and its growth was recorded. Then, the in vivo development of bacterial wilt was assessed following different elicitors application modes to both local tomato cultivars Tropimech and Caraïbo respectively sensitive and tolerant to this disease. Up to 100 ppm, BABA and ASM elicitors showed no antibacterial effect against R. solanacearum. However, these elicitors revealed a protective action against the development of bacterial wilt after inoculation of R. solanacearum. Compared to leaf treatment, the supply of elicitors to roots or successively to roots and leaves reduced the development of bacterial wilt by more than 50%. Both tomato varieties (Tropimech and Caraibo) expressed identical resistance levels facing Ralstonia solanacearum after BABA and ASM application. The elicitation of tomato plants could be an ecological approach for effective control of R. solanacearum. It is concluded that control bacterial wilt caused by R. solanacearum using ASM and BABA as elicitors of tomato defense. Two tomato cultivars that are Tropimech and Caraïbo respectively sensitive and tolerant to bacterial wilt were used. Tropimech which is the sensitive cultivar expressed identical resistance levels facing Ralstonia solanacearum after BABA or ASM application as Caraibo, the tolerant cultivar. Thus, ASM and BABA can be used in control to bacterial wilt of tomato. This technique can be considered as an ecological approach and an alternative to chemical control of bacterial wilt caused by R. solanacearum in tomato.

Biography of author(s)

Amari Ler-N’Ogn Dadé Georges Elisée
Laboratoire de Physiologie Végétale, UFR Biosciences, Université Félix Houphouët Boigny, 22 BP 582 Abidjan 22, Côte d’Ivoire.

N’guessan Aya Carine
Département de Biologie Végétale, UFR Sciences Biologiques, Université Péléforo Gon Coulibaly, BP 1328, Korhogo, Côte d’Ivoire.

Bomisso Edson Lezin
Laboratoire de Physiologie Végétale, UFR Biosciences, Université Félix Houphouët Boigny, 22 BP 582 Abidjan 22, Côte d’Ivoire.

Kouakou Tanoh Hilaire
Laboratoire de Biologie et Amélioration des Productions Végétales, UFR des Sciences de la Nature, Université Nangui Abrogoua, 02 BP 801 Abidjan 02, Côte d’Ivoire.

Ake Sévérin
Laboratoire de Physiologie Végétale, UFR Biosciences, Université Félix Houphouët Boigny, 22 BP 582 Abidjan 22, Côte d’Ivoire.

Kone Daouda
Laboratoire de Physiologie Végétale, UFR Biosciences, Université Félix Houphouët Boigny, 22 BP 582 Abidjan 22, Côte d’Ivoire.

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Physicochemical Properties of an Avian Protease Inhibitor | Chapter 06 | Recent Advances in Biological Research Vol. 2

Background: Ovomucoid is a serine proteinase inhibitor in the egg whites of all avian species at a concentration of about 10 mg/ml. The involvement of proteinases in a multitude of control functions in an organism has created an interest in their physiological inhibitors. Regulation of proteolytic activity in tissues is a critical requirement in the maintenance of homeostasis. Egg white proteins possess ACE-inhibitory activity & high radical-scavenging activity. The combined antioxidant and ACE-inhibitory properties of egg white hydrolysates, or the corresponding peptides would make a useful multifunctional preparation for the control of cardiovascular diseases. Proteases play key roles in several physiological processes, including intracellular protein degradation, bone remodeling, and antigen presentation, and their activities are increased in pathophysiological conditions such as, cancer metastasis and inflammation. They are also required for invasion by the microorganism. Four protease inhibitors have been identified in egg white namely, cystatin, ovomucoid, ovomacroglobulin (also known as ovostatin), and ovoinhibitor. Use of proteinase inhibitors in the treatment of certain diseases has renewed interest in their specificity and stability, both of which in turn depend on the tertiary structure of the inhibitor. Structural alteration to obtain molecules of desired properties requires knowledge of relationship between structure, function and stability.

Aims: Given its importance, in the present study duck ovomucoid was isolated and characterized for its physicochemical properties.

Methodology: Duck ovomucoid was isolated and characterized by its physicochemical properties. Analytical gel filtration (Sephacryl S-100 HR column) was used for purification, determination of molecular weight (MW), carbohydrate content and Stokes radius.

Results & Conclusion: The fluorescence emission spectrum was 302 nm, comparable to earlier reports. Stoke’s radius was found to be 2.91nm, and the value was comparable with white leghorn hen (Stoke’s radius 3.15nm). The extraordinary large value of stokes radius can be attributed to its high carbohydrate content which increases the hydration of the molecule. The inhibitor had the molecular weight of 29,300, and the carbohydrate content was 22%, the specific extinction coefficient of duck ovomucoid was found to be 5.82 at 279 nm and the Stoke’s radius was 2.91nm. The extraordinary large value of stokes’ radius of duck ovomucoid can be attributed to its high carbohydrate content which increases the hydration of the molecule resulting in large Stoke’s radius. Further functional studies on duck ovomucoid are required, as it has been reported that the chicken ovomucoid hydrosylates have some exceptional antioxidant, ACE inhibitory and metal chelating activities.

Biography of author(s)

Khushtar Anwar Salman
Department of Biochemistry, JN Medical College, A M U Aligarh, India.

Sharique Ahmed
Department of Allied Health, College of Health Sciences, University of Bahrain,Bahrain.

Indu Saxena
Department of Biochemistry, AIIMS, Jodhpur, India.

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Comparing the Susceptibility of Chironomus Striatapennis Larvae Exposed to Heavy Metals | Chapter 05 | Recent Advances in Biological Research Vol. 2

Fourth instar larvae of Chironomus striatapennis were collected from breeding aquarium under laboratory conditions and exposed for 96 hours to different doses of Pb, Cd and Hg for static bioassay to measure the LC50. Ten fourth instar larvae were placed in 100 ml beaker with 50 ml of each test solution. Larvae were exposed to six different concentrations, consisting of five trials. A control was also maintained wherein organisms were exposed to distilled water. Larvae were not fed during the toxicity tests. All beakers were free from tube forming materials. Data of mortality were subjected to probit analysis. Chi square was used to test for heterogeneity and the result was found to be significant (p<0.05) in all three metals. Results showed that sensitivity of larvae to metals was Hg> Cd >Pb. C. striatapennis showed noticeable response in LC50 study and was sensitive to low doses of heavy metals. Several secondary consumers have preferred this larva as their food. So unplanned industrialization may increase the level of heavy metals in the aquatic ecosystem which will accumulate slowly but definitely in different trophic levels and at the same time unusual death of these larvae may indirectly change the equilibrium of the aquatic ecosystem. Static bioassay of heavy metals revealed that larvae of Chironomus striatapennis was more sensitive to Hg than Cd and Pb respectively. It was also observed that LC50 values were less than standard permissible limit of these heavy metals. Unplanned industrialization may increase the level of heavy metals in the aquatic ecosystem which will accumulate slowly but steadily in different trophic levels and at the same time unusual death of these larvae may indirectly change the equilibrium of the aquatic ecosystem. As this larva is a preferred food for different secondary consumers, heavy metal toxicity may responsible for bio-accumulation of several heavy metals in chironomid larvae and make these larvae unsuitable for consumption.

Biography of author(s)

Dr. Susanta Nath
Associate Professor in the Department of Zoology, Government General Degree College Singur under the Department of Higher Education, Government of West Bengal, India.

Dr. Biplob Kumar Modak
Associate Professor in the Department of Zoology, Sidho Kanho Birsha University, Purulia, West Bengal, India.

Dr. Sudipta Das
Associate Professor of Zoology under West Bengal Education Service, India.

Rahul Podder
Department of Zoology, Sidho Kanho Birsha University, Purulia, West Bengal, India.

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Fatty Acid Methyl Ester Analysis of Olive Oil Degraded by Pseudomonas fluorescens and Enzymatic Characterisation of the Lipase | Chapter 04 | Recent Advances in Biological Research Vol. 2

Vegetable oils are harmful to the environment, like petroleum oils they produce similar environmental effects. Hence, it is a known fact that the olive oil causes the production of large quantity of olive mill waste (OMWs) as by-products to the environment. These by-products are harmful to the environment, both  terrestrial  and  aquatic.  Bacteria  are  very  useful  in  protecting  the  environment.  The  lipolytic activities  of  physiologically  diverse  bacteria  have  great  potential  to  degrade  oil  spills  in  the environment. Fatty  acids  are  the  major  components  of  lipids;  and  the  physical,  chemical  and physiological properties of a lipid class depend primarily on its fatty acid composition.There is need for extensive characterisation of the bacterium lipase for the treatment of vegetable oil-polluted sites. This work was carried out to preliminarily characterise the lipase of Pseudomonas fluoresens and to check  the  fatty  acid  composition  of  olive  oil.Gas  chromatography  method  can  be  used  for  the identification of microbiological degraded fatty acids in vegetable oils as methyl ester.Pseudomonas fluoresenswas screened for lipase production using standard methods. Temperature, pH, ion concentration (NaNO3and MgSO4), enzyme concentration, nitrogen concentration, substrate concentration,  time  course  and  agitation  speed  were  optimised  for  the  lipase  activity  as  well  as growth.Crude enzyme of Pseudomonas fluoresenshad the highest lipase activity and growth of 0.8 U/mL and 1.418 mg/mL respectively at room temperature, but when production was optimised higher activity 0.9 U/mL was seen  in the use of glucose as substrate. Agitation with the speed used did not support lipase  production  but  supported  growth  (1.998  mg/mL)  at  agitation  speed  of  100  rpm. Olive  oil degraded by  lipase of Pseudomonas fluorescens grown in two different  Mineral Salt Medium, was studied over 25 days. Olive oil was analyzed for fatty acids commonly present in olive oils which are Myristic,  Palmitic,  Stearic,  Oleic,  Linoleic,  Linolenic,  Behenic  and  Lignoceric,  which  have  specific carbon number and their values in approximate percentage are C14:0 (0.4), C16:0 (14.0), C18:0 (5.5), C18:1  (76.4),  C18:2  (3.4),  C18:3  (0.1),  C22:0  (0.1)  and  C24:0  (0.1)  respectively.  Oleic  acid percentage is high in olive oil which contained considerable amount of 76.4%.The reduction of fatty acid by Pseudomonas fluorescens after 20 days was 8.2% in the media used. Lipase from Pseudomonas fluorescens had potential for degradation of fatty waste. It could therefore be employed in environmental cleanup of oil spill site. Lipase production is generally influenced by the type and concentration of carbon and nitrogen sources, the culture pH, the growth temperature and the dissolved oxygen  concentration [1].Hence  monitoring the result of degradation patterns of the vegetable oil using fatty acid methyl ester analysis, by Pseudomonas fluorescens, showed that this organism has great potential for fatty waste degradation and as an extension in environmental clean up of oil spill site.

Biography of author(s)

Dr. B. M. Popoola
Department of Microbiology, University of Ibadan, Ibadan, Nigeria

Prof. Onilude A. A.
Department of Microbiology, University of Ibadan, Ibadan, Nigeria

Olateru Comfort T.
Department of Microbiology, University of Ibadan, Ibadan, Nigeria

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