Microbiological Quality of Raw Vegetables and Ready to Eat Products Sold in Abidjan (Côte d’Ivoire) Markets | Chapter 16 | Theory and Applications of Microbiology and Biotechnology Vol. 1

Vegetables are usually consumed raw. This implied best hygienic conditions from the harvest to the processing because of the gastro-enteritis that they could provoke. This study was conducted with the aim to appreciate microbiological quality of raw tomatoes, endives and ready-to-eat products sold in markets. Samples were taken randomly in two markets of Abidjan. A microbiological analysis was done in order to identify and enumerate fecal coliforms, Escherichia coli, Enterococcus and Pseudomonas. A decontamination treatment based on washing samples with running water and sodium hypochlorite solution I° chlorymetric was also applied to tomatoes and endives. The results indicated that, for tomatoes and endives, the average load was 1.5.104 CFU/g of Enterococcus, 1.3.103 CFU/g of Pseudomonas and 1.7.102 CFU/g of fecal coliforms. In ready-to-eat products, the load was 9.3.101 CFU/g for Enterococcus, l.03.101 CFU/g for Pseudomonas and 9.9.101 CFU/g for fecal coliforms. The disinfection with a sodium hypochlorite solution l° chlorymetric reduced Enterococcus and fecal coliforms load to 98% and Pseudomonas load to 97% as compared to the washing with running water in which Enterococcus was only reduced to 80%, fecal coliforms to78% and Pseudomonas to 73%. Escherichia coli were isolated in 28 samples as follow: 15 stumps from endives (54%), 10 stumps from tomatoes (36%) and 3 stumps from ready-to-eat products (10%). Results showed that before consumption, vegetables need to be washed, cleaned and disinfected. This will avoid sanitary hazard.

Author(s) Details

Dr. Julien Coulibaly-Kalpy
Food Safety Unit, Pasteur Institute of Côte d’Ivoire, 01 BP 490, Abidjan, Côte d’Ivoire.

Dr. Edith Adouko Agbo
Laboratory of Nutrition and Food Security, Nangui Abrogoua University, 02 BP 801, Abidjan, Côte d’Ivoire.

Professor Thomas Adjehi Dadie
Laboratory of Food Biotechnology and Microbiology, Nangui Abrogoua University, 02 BP 801, Abidjan, Côte d’Ivoire.

Professor Mireille Dosso
Food Safety Unit, Pasteur Institute of Côte d’Ivoire, 01 BP 490, Abidjan, Côte d’Ivoire.

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Response of Pseudomonas Species from Contaminated Soils to Selected Organic (Synthetic) Pesticides | Chapter 15 | Theory and Applications of Microbiology and Biotechnology Vol. 1

Growth response profile of three species of Pseudomonas isolated from pesticide contaminated soils within Uyo, Nigeria was studied using standard microbiological and analytical techniques. The ability of the isolates to tolerate varying concentrations of commercially available pesticides namely; Decis, DD force and Cyperforce was assessed over a 96 hour period. Selective enrichment cultures with graded concentrations of the pesticides were used to assay their growth response profile and the absorbance determined using CO75 digital colorimeter. The results showed that the Pseudomonas species differed biochemically. Their growth response at pesticide concentrations 0.0001, 0.001, 0.01, 0.1, 1 and 10% v/v differed significantly (P<0.05) at 24 hours interval for four days. At 10% concentration of Decis pesticide over 96 hour, the absorbance were 0.20, 0.23 and 0.30 for Pseudomonas from Agriculture research farm, hospital dumpsite and municipal waste dump site respectively. This ability therefore offers a veritable tool for use in the bioremediation and ultimate restoration of pesticide contaminated soils but however requires further evaluation.

Author(s) Details

Uduak U. Ndubuisi-Nnaji
Department of Microbiology, University of Uyo, Uyo, Nigeria.

Utibe A. Ofon
Department of Microbiology, University of Uyo, Uyo, Nigeria.

Godwin E. Udofia
Department of Microbiology, University of Uyo, Uyo, Nigeria.

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Potential Risk Factors and Prevalence of Infection of Helicobacter pylori in Nigeria | Chapter 14 | Theory and Applications of Microbiology and Biotechnology Vol. 1

Aim: Potential risk factors and prevalence associated with Helicobacter pylori (H. pylori) infection in apparently healthy children in Nigeria were studied.

Study Design: To investigate the current potential risk factors associated with recent prevalence of H. pylori in apparently healthy children in Nigeria.

Place and Duration of Study: The study was conducted in two Local Government Areas, Alimosho and Ajeromi, of Lagos State, Nigeria between March and September 2014.

Methodology: Seroprevalence status of 185 asymptomatic children made up of 93 males and 92 females, aged between 2-16 years were selected by randomized stratified sampling with descriptive questionnaire. Serum immunoglobulin G H. pylori antibody of the individual subjects was determined using DiaSpot H. pylori kit while fecal samples of same group were analysed for HpSAg using immunoassay test kit of Helicobacter pylori Stool Antigen (HpSAg).

Results: Of 185 children tested for H. pylori antigen, 134 (68.7%) and 51(26.2%) were classified as seropositive and fecal HpSA positive respectively. Highest rate of 40.0% and 34.6% of the children weighing between 21 and 40 kg were positive while 29.2% and 32.5% children of parents that were traders were positive to serum H. pylori antigen and fecal HpSA respectively. Only 12.4% and 14.1% children from artisan parents were positive but different age group have no association with the infectivity or prevalence of fecal H. pylori antigen (OR=0.67, CI=0.142-0.152). Significant higher percentage of seropositivity of 59.0% and fecal positivity of 55.7% was recorded among children from 5-8 people in a room (p>0.05), while Households with regular potable water supply have lower H. pylori seropositivity and fecal positivity of 11.9% and 7.6% compared with households that sometimes have water supply. The Households that never had water supply had highest number of seropositivity of 40.0% and 18.4%, respectively. Sewage nearness to kitchen indicates 30.8% and 28.7% H. pylori seropositive and fecal positivity rate among children.  

Conclusion: Paediatric H. pylori prevalence is highly associated with water borne infection and poor sanitary practices. There is need for achievable interventions and improvement in environmental sanitation.

Author(s) Details

F. O. Olufemi
Department of Veterinary Microbiology and Parasitology, College of Veterinary Medicine, Federal University of Agriculture, Nigeria.

Quadri Remi
Department of Microbiology, College of Biological Sciences, Federal University of Agriculture, Abeokuta, Nigeria.

P. A. Akinduti
Department of Veterinary Microbiology and Parasitology, College of Veterinary Medicine, Federal University of Agriculture, Nigeria.

S. A. Bamiro
Department of Physiology, College of Medicine, Lagos State University, Lagos, Nigeria.

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Prevalence of Urinary Tract Infection and Antibiotic Resistance Pattern in Pregnant Women, Najran Region, Saudi Arabia | Chapter 13 | Theory and Applications of Microbiology and Biotechnology Vol. 1

Urinary Tract Infection (UTI) is one of the commonest infectious disease in pregnancy, and in pregnancy we have very limited number of antibiotics to treat the UTI. This study was conducted on 151 patients who attended the gynecology clinic during the study period. Nineteen UTI proven cases of UTI were studied for prevalence of microorganism and sensitivity pattern against different antibiotics. Among the bacteria isolated, Escherichia coli (73.68%) and Staphylococcus aureus (10.52%) were the most prevalent Gram negative and Gram positive bacteria respectively. To know the resistance pattern of microorganism we used commercially available discs of different antibiotics. Gram negative bacteria showed more resistance as compared to Gram positive one. It is observed that the most effective antibiotic for Gram negative isolates is Ceftriaxone (87.5%), followed by Amoxicillin + Clavulanic acid (81.25%), Amikacin (75%), Cefuroxime (75%), Cefixime (68.75%) and Mezlocillin (62.5%). For the Gram positive bacteria, Ceftriaxone, Amikacin and Amoxicillin + Clavulanic acid were the most effective antimicrobials (100%). Multidrug resistance Gram negative bacteria were also tested for Extended-spectrum Beta Lactamase (ESBL), 35.71% of E. coli isolates were ESBL producer.

Author(s) Details

Dr. Ali Mohamed Alshabi
Department of Clinical Pharmacy, College of Pharmacy, Najran University, Najran, Saudi Arabia.

Dr. Majed Saeed Alshahrani
Department of Obstetics and Gyneocology, Faculty of Medicine, Najran University, Najran, Saudi Arabia.

Dr. Saad Ahmed Alkahtani
Department of Clinical Pharmacy, College of Pharmacy, Najran University, Najran, Saudi Arabia.

Mohammad Shabib Akhtar
Department of Clinical Pharmacy, College of Pharmacy, Najran University, Najran, Saudi Arabia.

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Biotreatment of Crude Oil Contaminated Soil | Chapter 12 | Theory and Applications of Microbiology and Biotechnology Vol. 1

Biodegradation of hydrocarbons by microorganisms represents one of the primary mechanisms by which petroleum and other hydrogen pollutants are eliminated from the environment. This work was carried out on the effect of microorganisms on the biotreatment of oil in crude oil contaminated soil.

Microorganisms were isolated from two experimental soil samples contaminated with Bonny Crude and normal uncontaminated soil as a control over a period of seven months. The microbial as well as the physico-chemical parameters of the soil samples were all analyzed using standard methods. Changes in total petroleum hydrocarbon level were measured appropriately. Treatments used were the microbial isolates.

Forty-four microorganisms were isolated from the contaminated soils and identified as species of Pseudomonas (7), Flavobacterium (6), Bacillus (8), Proteus (4), Klebsiella (1), Pencillium (5), Aspergillus (7), Fusarium (3), Trichypton (2) and Neurospora (1). Ten of the forty-four isolates had ability to degrade crude oil in the laboratory. On contamination a value of 1.0X105 cfu/g in microbial counts were obtained followed by a subsequent increase in population levels after a period of 2months with a value of 1.0X106 cfu/g. Oil application to the soil resulted in an increase in total petroleum hydrocarbon from 0.31 ppm to 5.53 ppm; organic matter from 0.41% to 7.34%; available phosphorus from 1.75 ppm to 2.84 ppm. The treatment measures all showed progressive decrease in oil concentration in the soil. Mixture of bacterial and fungal isolates as a treatment measure proved to be more favourable above all others, it brought the concentration from 5.53 ppm to 0.31 ppm after a period of 5 weeks of treatment, which is same value with the normal soil (uncontaminated).

Species of Pseudomonas, Bacillus, Flavobacterium, Proteus, Klebsiella, Penicillium, Aspergillus, Fusarium, Trichyphyton and Neurospora had potential for the degradation of bonny crude oil. They could therefore be employed in environmental cleanup of petroleum spill site.

Author(s) Details

B. M. Popoola
Ajayi Crowther University, Oyo, Oyo State, Nigeria.

A. A. Olanbiwonninu
Ajayi Crowther University, Oyo, Oyo State, Nigeria.

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Exploration of Sulfate Reducing Bacteria from Polluted Waters | Chapter 11 | Theory and Applications of Microbiology and Biotechnology Vol. 1

Sulfate reducing bacteria (SRB) was successfully isolated from Estuary Dam in Suwung Denpasar, Indonesia. This estuary catches highly polluted water from Badung River which runs across and hence carries pollution due to waste disposal from Denpasar City. SRB was studied in detail for their ability to reduce sulfate to sulfide with organic material as an oxidizing agent. SRB exploration of the estuary ecosystem of the contaminated dam was accomplished through isolation, selection and characterization of the isolates obtained. The result of this study found superior SRB named DPS 1711, DPS 1705 and DPS 1703. The bacteria have the ability to grow at pH 3, room temperature and uses compost as organic substrate. This ability is an important factor for the application of isolates in the treatment of acid mine waste. Isolates have optimum optical density under the pH range of 4 to 7 and the best at pH 5 have a growth rate profile at a temperature range of 25 to 40°C. The isolates observed were Gram-negative stem, motile bacteria which only grow in anaerobic condition. Physiological-biochemical characterization showed the three isolates, namely DPS 1703, DPS 1705 and DPS 1711 were SRB groups identified as Desulfotomaculum orientis.

Author(s) Details

Dr. W. Budiarsa Suyasa
Bioremediation Research Division, Udayana University, Denpasar Bali, Indonesia.

Iryanti E. Suprihatin
Bioremediation Research Division, Udayana University, Denpasar Bali, Indonesia.

G. A. Dwi Adi Suastuti
Bioremediation Research Division, Udayana University, Denpasar Bali, Indonesia.

G. A. Sri Kunti Pancadewi
Department of Environmental Chemistry, Udayana University, Bukit Jimbaran, Bali, Indonesia.

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Phytochemical Analysis and Antibacterial Efficacy of Mentha piperita (L) Ethanolic Leaf Extract against Clinical Isolates of Uropathogens | Chapter 10 | Theory and Applications of Microbiology and Biotechnology Vol. 1

Aim: The present study was designed to evaluate the antibacterial activity of Mentha piperita (L) leaf extract against clinical isolates of urinary tract infections.

Introduction: M. piperita L. (Peppermint) is a strongly scented herb belonging to family Lamiaceae. The plant is stimulant, aromatic and used for headache, vomiting and allaying nausea. In India the leaves are used to relieve sore throat. The most common form of bacterial infections is urinary tract infections (UTIs). They affect people of all age groups throughout their lifespan.

Methodology: The M. piperita ethanolic extract (MPEE) was prepared by cold maceration. The presence of phytoconstituents was determined using standard protocols. Clinical isolates of UTI pathogens such as Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa were isolated from urine samples and identified by biochemical tests. The antibacterial property was determined by agar well diffusion method.

Results and Discussion: The preliminary phytochemical screening revealed the presence of amino acids, carbohydrates, alkaloids, proteins, glycosides, steroids, tannins and flavonoids. MPEE exhibited pronounced antibacterial activity against tested microorganisms. The extract exhibited antibacterial activity at 1000 µg concentration against S. aureus (21.50±1.22 mm), E. coli (19.33±0.81 mm), P. aeruginosa (15.33±1.69 mm) from high to low respectively. The minimum inhibitory concentration was ranged from 62.5 to 125 mg/ml.

Conclusion: The results of antibacterial studies confirm that MPEE was found to contain bioactive constituents that exhibited notable antibacterial activity. However, further isolation and characterization of phytoconstituents will be needed to evaluate the antimicrobial activities against a wider range of microbial pathogens.

Author(s) Details

Dr. Sowjanya Pulipati, B. Pharm, M. Tech (Bio-Tech), Ph.D
Vignan Pharmacy College, Vadlamudi- 522213, Guntur, Andhra Pradesh, India.

Sai Koushik Oruganti
Master of Medical Biotechnology, University of Windsor, Canada.

P. Srinivasa Babu, M. Parm, PhD
Vignan Pharmacy College, Vadlamudi- 522213, Guntur, Andhra Pradesh, India.

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The Antimicrobial Effects of Secondary Metabolites of Anguillan Fungi | Chapter 09 | Theory and Applications of Microbiology and Biotechnology Vol. 1

Introduction: Many drugs have been isolated from fungal species. This study aims at identifying fungal species isolated in Anguilla to determine the antimicrobial effect of their secondary metabolites from pure culture against Staphylococcus aureus by demonstrating the presence of a zone of inhibition in the culture plate.

Methods: Samples were cultured and sub-cultured to isolate pure culture on potato dextrose yeast agar (PDYA) and chosen for further studies by the presence of surface exudates. Those cultures that produced copious amounts of surface exudates were examined for antimicrobial effects by further testing.

Results: Antimicrobial testing of MB0725C (P. digitatum) samples did not result in any evidence of antimicrobial properties. However, MB0725A (P. chrysogenum) punch biopsy-agar overlay produced an 11 mm ZOI, whereas crude exudates testing resulted in a 27 mm ZOI. Crude culture filtrate of potato dextrose broth (PDB) did not result in any ZOI for either MB0725A or C. Sensitivity testing on samples collected from Yeast Extract Lactose Broth (YELB) on Day 3, Day 6, Day 9 and Day 12 resulted in ZOI of 11 mm, 13, 15 and 17 mm respectively. The change in pH for MB0725A liquid culture in PDB versus YELB was significantly different (N=12, P<0.0001).

Conclusion: MB0725A was an excellent producer of surface exudates and further experiments showed that its secondary metabolites had antimicrobial effects against Staphylococcus aureus using Kirby-Bauer sensitivity testing.

Author(s) Details

Michael Bennardo
Department of Medical Microbiology and Immunology, Saint James School of Medicine, BWI, Anguilla.

Adekunle Sanyaolu
Federal Ministry of Health, Abuja, Nigeria.

Subhajit Dasgupta
Department of Medical Microbiology and Immunology, Saint James School of Medicine, BWI, Anguilla.

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Atypical Manifestation in Infection by Methicillin-Resistant Staphylococcus aureus Carrier SCCmec IV and Panton-Valentine Leukocidin-Producer in Experimental Sepsis Model | Chapter 08 | Theory and Applications of Microbiology and Biotechnology Vol. 1

Staphylococcus aureus is considered an infectious agent of great clinical importance, responsible for many different types of infection. Strains of methicillin-resistant Staphylococcus aureus (MRSA), Panton-valentine leukocidin producers, are considered more invasive, presenting clinical sequelae related to abscesses and infection in skin and soft tissues. The use of invasive techniques in hospital environment, such as the introduction of intravascular catheter in immunocompromised patients, has contributed to this microorganism spreading through the bloodstream, causing bacteremia, necrotizing pneumonia and increasing the number of septic patients in intensive care units with high mortality. In this report, atypical infections in Swiss mice using experimental model of sepsis was presented.

Author(s) Details

Giorgio Silva-Santana
Department of Pathology, School of Medicine, Fluminense Federal University, Rio de Janeiro, Brazil and Laboratory Academic Rodolfo Albino, Fluminense Federal University, Rio de Janeiro, Brazil.

Kátia C. Lenzi-Almeida
Department of Pathology, School of Medicine, Fluminense Federal University, Rio de Janeiro, Brazil and  Environmental Science and Conservation Department, School of Medicine, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.

Vânia G. S. Lopes
Department of Pathology, School of Medicine, Fluminense Federal University, Rio de Janeiro, Brazil.

Fábio Aguiar-Alves
Department of Pathology, School of Medicine, Fluminense Federal University, Rio de Janeiro, Brazil and Laboratory Academic Rodolfo Albino, Fluminense Federal University, Rio de Janeiro, Brazil.

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Endophytic Microbiome Population from Moringa oleifera Leaves Collected in Three Localities at Pernambuco State, Northeast Brazil | Chapter 07 | Theory and Applications of Microbiology and Biotechnology Vol. 1

Aims: This work investigated the population density of endophytic microorganisms from Moringa oleifera leaves collected in three localities at the State of Pernambuco (northeastern Brazil): Urban (campus from the Federal University of Pernambuco, UFPE) and forest (botanical garden) areas at Recife city and an urban area (industrial district) at Caruaru city.

Place and Duration of Study: Department of Antibiotics and Department of Biochemistry from Federal University of Pernambuco, during four years.

Methodology: Sodium hypochlorite was used to disinfect the leaves, which were macerated in PBS buffer and separately sowed on seven culture media supplemented with antibacterial or antifungal agents.

Results: The majority of endophytes isolated were bacteria and the highest density was found in leaves from the forest area. Regarding to fungi isolation, there was no statistical difference between the density in leaves from UFPE campus and the botanical garden while no fungal isolates was obtained from leaves collected in Caruaru. The highest diversity of endophytes was found in the leaves from the botanical garden, with 111 different isolates. A total of 71, 94 and 50 bacterial isolates were obtained from leaves of UFPE campus, botanical garden and Caruaru, respectively. The number of fungal isolates were 17 (UFPE campus) and 12 (botanical garden).

Conclusion: In conclusion, the methodology employed in this work was effective for the isolation of endophytes; it is still worth to mention that climatic and geographical conditions may interfere in density and diversity of endophytes from M. oleifera leaves.

Author(s) Details

Igor Felipe Andrade Costa de Souza
Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420, Recife, Brazil.

Thiago Henrique Napoleão
Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420, Recife, Brazil.

Thales Henrique Barbosa de Oliveira
Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420, Recife, Brazil.

Kêsia Xisto da Fonseca Ribeiro de Sena
Departamento de Antibióticos, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420, Recife, Brazil.

Patrícia Maria Guedes Paiva
Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420, Recife, Brazil.

Janete Magali de Araújo
Departamento de Antibióticos, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420, Recife, Brazil.

Luana Cassandra Breitenbach Barroso Coelho
Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420, Recife, Brazil.

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