Production and Characterization of cellulase enzyme from Aspergillus species using Cyperus esculentus

CHAPTER ONE/INTRODUCTION

Biotechnology has the ability to create a vast variety of products to satisfy different human needs. Enzyme technology, a subfield of biotechnology, has developed and is still developing new processes to produce both bulk and high value-added products using enzymes as biocatalysts in order to meet the demands for things like food (bread, cheese, beer, vinegar), fine chemicals (such as amino acids, vitamins), and pharmaceuticals. Enzymes may be used for analytical and diagnostic reasons in addition to cleaning and environmental procedures.
The advancement of enzyme technology in academia and industry has been and will continue to be driven by the improvement of current production processes to produce new goods from novel raw materials, such as biomass, or by the development of new, better products, processes, and services to meet these needs. These strategies will be used to create innovative, sustainable goods and processes. In 1987, the World Commission on Environment and Development (WCED) created the idea of sustainability to promote the necessary “development that fulfills the demands of the present without sacrificing the capacity of future generations to satisfy their own needs.” The Cartagena Protocol on Biosafety to the Convention on Biological Diversity, an international agreement that regulates the transfer of living modified organisms (LMOs) made possible by modern biotechnology from one nation to another, presently includes this description. It was approved on September 11, 2003, and on January 29, 2000, it was added to the Convention on Biological Diversity. 160 states have now approved it.
Biochemical catalysts known as enzymes are regarded as gifts from nature. Inside the cells, they operate like protein robots, speeding up biological processes without changing them (Lanka and Latha, 2015). Scientific research, cosmetic manufacturing, medical diagnostics, chemical analysis, medicinal uses, and industrial catalysis all make extensive use of enzymes (Sharma et al 2001b).

These beneficial compounds were created by using the cellulase enzyme to bioconvert lignocellulosic material. The local economy, ecology, and safety of the country’s energy supply would all gain from the production of biobased goods and bioenergy from less costly renewable lignocellulosic materials (Zhang, 2008). Numerous fungi generate the enzymes needed to convert proteins and polysaccharides into digestible sugars and amino acids. These enzymes are essential to the economy. Submerged fermentation (SmF), which often uses highly adapted and genetically altered bacteria, is used to create around 90% of all commercial enzymes. Thanks to SmF technology, solid state fermentation offers an unsurpassed edge in this sector (SSF). It’s noteworthy to note that recent studies on SSF of fungi, yeasts, and bacteria have shown a range of metabolic activities in both solid state and submerged fermentation environments.
Enzymes called cellulases break down the -1,4 bonds in cellulose strands. They are made by animals, plants, bacteria, protozoa, fungus, and bacteria. Different groups of catalytic modules of cellulases have been identified based on their crystal structures and amino acid composition (Henrissat, 1989). Cellobiohydrolases (CBHs), exoglucanases, and -glucosidase are the three main cellulase types that contribute to the whole cellulose hydrolysis process in nature. Microorganisms must produce these cellulases in order to hydrolyze and metabolize insoluble cellulose, maybe with the exception of BG, which is either free or bound to the cell surface.
Despite the fact that only 2% of the world’s microorganisms have been identified as sources of enzymes, microbial enzymes have a number of advantages over enzymes derived from plants and animals, including a higher potential for catalytic diversity, higher productivity in a short amount of time, ease of genetic manipulation/optimization, independence from seasonal fluctuations, and rapid growth of the producing microorganism (Wiseman, 1995)

Influence of organic fertilizers on growth and yield of pawpaw

The papaya, scientifically known as Carica papaya L., is widely regarded as one of the healthiest and tastiest fruits in the world. To all intents and purposes, it is a tropical fruit, and it is grown for commercial purposes in tropical and subtropical regions (Yadava et al., 1990). It is widely held that the disease originated in Mexico and subsequently spread to virtually all of the tropical regions of the world. It is an excellent source of protein, carbohydrates, fiber, vitamins A and C, beta-carotene, and minerals such as iron, phosphorous, and potassium (Mamta et al., 2017). The papaya is most commonly found in fresh-markets, but it is also used in beverages, jams, candies, pectin, and crystallized forms of the fruit. Cooking with green fruit is similar to cooking with other vegetables. Leaves have a long history of use in traditional medicine for the treatment of a wide variety of conditions, including malaria, dengue fever, jaundice, immune modulatory and antiviral activity, and more (Singh et al., 2020). It is utilized in a number of important industrial processes, most notably in the production of the enzyme pepsin (EI Moussaoui et al., 2001), which shares many of the same characteristics as gastric pepsin. It is one of the most popular fruits in Bangladesh, despite the country’s relatively low production of papaya in comparison to that of other countries around the world (Chowdhury et al., 2008). According to Singh et al. (2012), inadequate nutrient management is one of the primary factors contributing to the low productivity of papaya. The papaya tree produces flowers and fruit continuously throughout the year; however, there is very little information available regarding the use of organic farming methods in the cultivation of this crop (Reddy et al., 2010). It is a crop that requires a lot of nutrients and responds well to fertilizer. There is a possibility that the growth yield of papaya can be affected by both organic and inorganic fertilizers.
However, the use of inorganic fertilizers without discretion alters the soil’s physical, chemical, and biological properties, and it also creates a problem for the environment and a health hazard due to the toxic residual effects of the fertilizers. Some of these factors included an increase in the incidence of pests, diseases, and weeds, all of which had an impact on productivity. On the other hand, organic fertilizers like cow dung, vermicompost, poultry manure, FYM, and mustard oil cake are helpful in improving the texture, structure, water-holding capacity, aeration, and microbial activities of soil (El-Shakweer et al., 1998).

According to the findings of a number of studies, organic farming methods not only contribute to the protection of the natural environment by reducing the amount of soil erosion that occurs, but they are also the most environmentally friendly method of crop production (Ojeniyi, 2000; Maritus and Vleic, 2001). In addition, not only are crops grown organically more nutritious, but they also eliminate the possibility of pesticide residues being passed on to consumers, which helps to ensure the safety of food. In addition, the demand for products that are produced organically is growing each and every day. In light of this, there is an urgent requirement for an increase in the production of papaya using organic methods. In light of this, the purpose of the current study was to investigate the effects of organic and inorganic fertilizers on the growth, yield, and physiochemical properties of papaya, as well as to standardize the doses for the purpose of maximizing papaya yield.

The investigation of unconventional oils derived from tigernut has been the primary focus of this work. This is because the development of forest species results in either a small or nonexistent crop. The objective of this study was to evaluate the physico-chemical parameters (acidity, peroxide, iodine, and saponification values), as well as the fatty acid profile, tocols, and sterols of vegetable oils obtained from Cyperus esculentus that were produced in Benin. The findings of the research indicate that Cyperus esculentus has a lipid potential that ranges between 27 and 29%. The quality indexes that were obtained, such as acidity (1%), saponification (178 mg KOH / g oil), peroxide (9 O2/kg-oil meq), and iodine (73-80 mg of iodine g-oil), were all in accordance with recognized norms of appreciation of the quality of alimentary oils. The fatty acid profile was led by oleic acid, which comprised 63% of the total, followed by palmitic acid, which comprised 16% of the total, linoleic acid, which comprised 9-13% of the total, and stearic acid, which comprised 7-10% of the whole. Oleic acid was the most abundant of the fatty acids. The concentrations of arachidic acid (less than one percent), palmitoleic acid (less than two percent), and myristic acid (less than one percent) were all quite low (less than one percent). Because these vegetable oils contained a relatively high proportion of unsaturated fatty acids, they were suitable for use as dietary supplements (more than 70 percent). The field of nutrition devotes a lot of attention to discussing this subject. In addition to total protein (MS 9%), starch (33-40% DM), and total sugar (21-26% DM), the findings of this study indicate that oilcake contains a significant amount of the minerals nitrogen N (1.5%), phosphorus P (0.27%), and potassium K (1%), all of which point to their potential application as animal feed. It was determined that the unsaponifiable components of vegetable oil, such as sterols and tocols, have an edible concentration of 63 and 4000 mg/100 g, respectively, through the application of the LC-MS technique. This allowed for the possibility of their use in cosmetics.

Abstract

The present study is aimed to investigate the potential effects of Nauclea latifolia leaves extracts on seven sh pathogens: Escherichia coli, Staphylococcus aureus, Streptococcus iniae, Bacillus subtilis, Pseudomonas aeruginosa, Samonella typhi and Aeromonas hydrophila using agar well diffusion assay. The phytochemical screening and minimum inhibitory concentration of ethanolic and methanolic extracts of N. latifolia were determined using standard methods. Data were analyzed using ANOVA at P=0.05. Ethanolic and methanolic extracts of N. latifolia leaves exhibited antibacterial activity against all the pathogens investigated. Ethanolic extracts of N. latifolia had better antibacterial activity when compared to the methanolic extracts. No antibacterial activity was recorded in the negative control (distilled water) while antibacterial activity was recorded in the positive control (chloramphenicol). Agar well diffusion assay for antibacterial activity yielded the inhibitory zone of 9.18±0.04mm to 27.50±0.25 mm diameter for ethanolic and methanolic extracts of N. latifolia leaves respectively. The phytochemical screening for metabolites indicated the presence of saponins, alkaloids, avonoids, glucosinolates and protein and amino acids while phenol and tannin were not detected. The minimum inhibitory concentration of methanolic and ethanolic extracts of N. latifolia leaves were 450μg/ml and 900μg/ml respectively. The results indicated that leaves extracts of N. latifolia express better antibacterial effect against multi – drug resistant bacteria and it can be used as a potential source for various biomedical applications. Keywords: Fish pathogens, Nauclea latifolia, Antibacterial, Phytochemical screening, Clarias gariepinus

ISOLATION AND IDENTIFICATION OF KLEBSIELLA ON SEA FOOD (catfish, prawns, Titus fish, shrimps, periwinkles)

INTRODUCTION

Klebsiella pneumoniae is an important opportunistic pathogen that causes a variety of infectious diseases in humans, including septicaemia, liver abscesses, diarrhea, and pneumonia (Bi and Xu, 2013; Cao et al., 2014; Guo et al., 2017). It is a well-known hospital-acquired pathogen and associated with increased patient morbidity and mortality (Brisse et al., 2009; Cabral et al., 2012). In addition to the clinical environment, K. pneumoniae is frequently found in foods including raw vegetables, powdered infant formula, meat, fish, and street foods, and has been considered as an important food-borne pathogen (Haryani et al., 2007; Sun et al., 2010; Puspanadan et al., 2012; Overdevest et al., 2014; Kim et al., 2015; Davis and Price, 2016). In powdered infant formula, K. pneumoniae is included in the hazard identification category “B” according to the FAO and WHO guidelines on microorganisms (FAO-WHO, 2004). In recent years, an increasing number of food-borne outbreaks caused by K. pneumoniae have been reported in different countries (Calbo et al., 2011; Tambekar et al., 2011; Zhou et al., 2011; Bi and Xu, 2013; Yu and Zhou, 2013).

K. pneumoniae can express a variety of virulence factors including capsules, endotoxins, siderophores, iron-scavenging systems, and adhesins, which have been shown to play important roles in its pathogenesis. Capsule is an important virulence factor, which is involved in at least two pathogenic mechanisms: (1) protection of the bacteria from phagocytosis, and (2) direct inhibition of the host immune response (Kang et al., 2015). Some capsular (K) types, particularly K1, K2, K54, K57, K20, and K5, are often associated with community-acquired invasive pyogenic liver abscess syndrome, septicemia, and pneumonia (Fang et al., 2007; Siri et al., 2011; He, 2012). K1, K2, K20, K54, and K57 are highly virulent in experimental infections in mice and are often associated with severe infections in humans and animals (Turton et al., 2008; Yu et al., 2008; Cheng et al., 2015; Wei et al., 2016). K2 and K5 are frequent causes of metritis in mares and are associated with community-acquired pneumonia (Brisse et al., 2009). K3 is generally associated with rhinoscleroma (He, 2012). Capsule typing is currently the most widely used technique for typing K. pneumoniae isolates and exhibits good reproducibility in differentiating clinical isolates (Siu et al., 2011). Several PCRs targeting the wzy genes have been developed for capsule typing of K. pneumoniae (Turton, 2010; Cheng et al., 2015). Other virulence factors such as the rmpA gene (regulator of mucoid phenotype A); allS gene (encoding the activator of the allantoin regulon, associated with allantoin metabolism); endotoxin-related genes wabG, uge, and wcaG; iron acquisition system-related genes iucB, iroNB, ybtA, and kfuBC; adhesin gene fimH (type I fimbriae); and ureA gene (α-subunit of the urease, invasin related) are also believed to be involved in virulence processes (Brisse et al., 2009; Turton, 2010; El et al., 2013; Calhau et al., 2014). The detection of such virulence factors is important in understanding the pathogenic characteristics of K. pneumoniae isolates and enhancing our knowledge of the health risks posed by this pathogen.

The emergence of antimicrobial resistance in Klebsiella spp. isolates is of great concern worldwide in human medicine (Hu et al., 2013). Multidrug-resistant K. pneumoniae strains have been isolated from different samples (Nawaz et al., 2012; Falomir et al., 2013; Guo et al., 2016; Yaici et al., 2017). Dietary intake is one of the primary routes for the introduction of antibotic-resistant bacteria and their genes into the human digestive tract. Consumption of specific food categories might influence gut antibiotic resistance gene diversity (Milanović et al., 2017). Moreover, such bacteria may transfer antibiotic resistance determinants to other pathogenic bacteria (Machado et al., 2008). Therefore, surveillance and monitoring of drug-resistant bacteria in foods is important for implementing targeted control strategies and selecting effective drugs for treatment.

Molecular typing is a useful tool for determining the genetic relationships of food-borne bacteria and identifying probable sources of infections. This is particularly important in endemic and epidemic nosocomial outbreaks of K. pneumoniae infections to improve the management of such outbreaks. A variety of methods have been used for K. pneumoniae typing, including pulsed-field gel electrophoresis (PFGE), enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR), randomly amplified polymorphic DNA (RAPD), and (GTG)₅ oligonucleotide PCR (Haryani et al., 2007; Ryberg et al., 2011; Barus et al., 2013; Sachse et al., 2014). The ERIC, RAPD, and (GTG)₅-PCR assays are relatively simple and cost-effective methods that have been successfully used for genotyping K. pneumoniae isolates from various sources (Ryberg et al., 2011; Barus et al., 2013).

In recent years, the number of food-borne illness outbreaks caused by K. pneumoniae has increased. However, until now, limited information has been available on the characteristics of K. pneumoniae isolated from foods. The purpose of the present study was to determine the biotypes, serotypes, virulence genes, and antimicrobial resistance patterns of food isolates and to further analyse their genetic diversity using ERIC-PCR and (GTG)₅-PCR molecular typing.

microbial isolation and biosafety assessment of beef components of three commercial sausage roll brands in Umuahia metropolis

MICROBIAL CONTAMINATION OF WELL WATER WITHIN MAKURDI METEROPOLIS

CHAPTER ONE/INTRODUCTION

Background to the Study

The inadequate supply of clean drinkable waters and the frequent pollution of existing supplies create very grave health problems for people in developing countries like Nigeria. Water serves as a vehicle for the transmission of diseases like typhoid fever, cholera, rashes, diarrhea, dysentery, Gastro-enteritis etc which have contributed immensely to the stagnation of the economic development of some of these nations (Okoufu et al., 1990).

In Nigeria, several studies have been carried out in urban areas on ground water contamination (Awalla, 2002; Akpabio and Ebong, 2004; Egbulem, 2003; Ahmed, 2003; Adebayo and Bashire, 2002). The uniform findings from these studies is that groundwater is highly contaminated and clinically unsafe for human consumption. With increasing population densities of urban areas and the corresponding increase in demand for social amenities, it has become increasingly difficult to meet all the water requirements in quantity, regularity and quality. The public water supply is generally inadequate and in most cases inaccessible, the supply is intermittent and unreliable thus resulting into high degree of population densities and concentration of socio-economic activities, it has become increasingly difficult to meet all the water requirements.

The public water supply is generally inadequate and in most cases inaccessible, the supply is intermittent and unreliable, thus resulting into high dependency on unsafe supplementary sources such as streams, hand dug wells and ponds (Kakulu and Osibanjo, 1992; Olajire and Imeppeoria, 2001; Nnodu and Ilo, 2002; Owuama and Uzoije, 2005; Ocheri, 2006; Maxwell et al., 2010). Concern over the quality of water harnessed especially from the hand dug wells have received wide attention among researchers (Ovrawah and Hymore, 2001; Ehinola and Coker, 2002; Nnodu and Ilo, 2002; Ogunbadewa, 2002; Omofonmwam and Eseigbe, 2009). Consistent in their findings is that water from hand dug wells are polluted through physical processes, geochemistry of the environment and anthropogenic activities. Consequently consumers of such waters are exposed to series of health risks. This study investigated the bacteriological characteristics of well water in the study area.

Statement of the Problem

Water has been recognized as a potential carrier of diseases and in line with frequent outbreak of typhoid fever, cholera and other water borne diseases; it is pertinent that water be adequately checked and possibly treated particularly well water which is the focus of this work.

Aim and Objectives of the Study

The main objective of this study is to investigate the Microbial Contamination Of Well Water Within Makurdi Meteropolis

The study made up of the following specific objectives;

1. To investigate the bacteria commonly present in well water in Makurdi
2. To find out the extent of microbial contamination in well water in different areas of Makurdi
3. To identify the source of the microbial contamination of well water within Makurdi Metropolis.

Scope of the Study

The study is limited to Makurdi metropolis.Both physico-chemical and bacteriological parameters are considered. The physico-chemical parameters considered are; Turbidity, Potential Hydrogen (pH), Total Dissolved Solid, Iron, Colour, Conductivity, Odour, Chloride, Fluorides and Nitrate while the bacteriological parameters include: Coli- form and Escherichia Coli (E-Coli).

Significance of the Study

The study is significant because it will aid in policy formulation by relevant government agencies and institutions in the formulation of policies regarding water production and distribution in Makurdi metropolis. The study will be of immense importance to members of the public as it can go a long way in enlightening them on the dangers of drinking unsafe and unhygienic water without treatment. The study will act as a guide to scholars and researchers who may wish to carry out research in a similar area especially as regards water quality.

The potential of actinomycetes and fungi to degrade spent oil from motor mechanic site soil

Bacteriological and molecular analysis of rifampin and isoniazid resistant strains of Mycobacterium tuberculosis isolated in Kaduna state Nigeria

ABSTRACT

COMPARATIVE STUDIES ON THE DIAGNOSIS OF SALMONELLA TYPHI USING RAPID KIT AND WIDAL

ABSTRACT

Typhoid fever continues to be a public health issue in most developing countries.Typhoid Fever ranks among the first twenty causes of outpatient illness. Among the numerous challenges in addressing the disease burden of Typhoid is the quest to secure a more reliable and standardized laboratory diagnoses of Salmonella infections to be able to win the surveillance battle on Typhoid fever. The purpose of this study was to compare the specificity and sensitivity of Typhidot and Widal serological tests for the detection of typhoid fever in Cameroon. A prospective hospital based longitudinal study was conducted by gathering samples of 292 potential patients who showed features of Typhoid infection. Adopting a purposive sampling technique, patients who met the inclusion criteria were conveniently sampled and enrolled. Blood culture was used as the standard protocol after which sensitivity, specificity and positive predictive values were compared between Typhidot and Widal serological test kits. Data was analyzed using SPSS version 20 to process after which results were presented descriptively. Chi square test of association was performed for categorical variables .The study on 292 individuals recorded sensitivity of widal (95%) and typhidot (85.8%) with the specificity of widal (54.0%) and Typhidot (90.1%). This observation indicates Widal test is much more sensitive than the typhidot in the study area. The study established that both tests could be equally, sensitive and specific in diagnosing typhoid since they both had an  equal accuracy (98.0) making them suitable for rapid diagnosis. The study found high detection of S. typhi by blood culture diagnosis, with S. paratyphi and S. typhimurun occurring least. This study has implication for further study to be carried out for confirmatory purposes as well as to determine prevalence of infection and antigenic variants that were not captured in the present study.