Theory and Applications of Microbiology and Biotechnology Vol. 3

The toxicological bioassay of petroleum products (industrial and local ‘kpo-fire’ refined Kerosene) in tri-aquatic ecosystem (marine, brackish and freshwater) using pollution bio-monitor Pseudomonas sp. were investigated. The study employs experimental examination and statistical analysis of data and interpretation. It was designed to evaluate the different kerosene concentration and the duration of exposure that could cause potential toxicological effect on Pseudomonas sp. in tri-aquatic ecosystem which was used as indices to access level of pollution. Standard microbiological techniques were used; toxicity procedure were applied using local and industrial refined kerosene; prepared at concentrations of 1.625%, 3.25%, 6.5%, 12.5% and 25% in fresh, brackish and marine water; total of 36 different microcosms. These were tested with Pseudomonas sp. for 0, 4, 8, 12 and 24 h separately for each toxicant. The cultures were incubated at 35°C for 24 hours. The median lethal concentration (LC₅₀) was employed to compute the toxicities of the different toxicants on the test organism. The results specify that percentage (%) logarithm of mortality of Pseudomonas sp. increases with increased toxicants concentration and exposure time. The pollution bio-monitor Pseudomonas sp. demonstrated sensitivity to the toxicity of local and industrially refined kerosene. The sensitivity showed variations, toxic level decreased in the following order (noting that the lower the LC₅₀, the more toxic the toxicants): Industrial refined kerosene in fresh water (18.79%) > Industrial refined kerosene in brackish water (20.81%) > Local refined kerosene in brackish water (21.47%) > Industrial refined kerosene in marine water (22.66%) > Local refined kerosene > (24.25) > Local refined kerosene in marine water (24.94%). Using the Pollution/Toxicity Bio-monitoring evaluation Chart; Local refined kerosene in marine, brackish and freshwater were ‘Toxic [High], Industrial refined kerosene in marine water was ‘Toxic [High]’ while Industrial refined kerosene in brackish and freshwater were ‘Toxic [very High]’.

Conclusion: The study showed that industrial refined kerosene in fresh water (LC₅₀ = 18.8%) has   the highest toxicity strength while local refined kerosene in marine water (LC₅₀ = 24.92%) has the least toxicity strength on Pseudomonas sp. in the tri-aquatic ecosystem. These results show that local and industrial refined kerosene can inhibit the growth of Pseudomonas sp. in an aquatic ecosystem; noting that Pseudomonas sp. is one of the most effective biodegrading bacteria in ecological biogeochemical cycles, pollutant removal/remediation and a key pollution bio-monitoring. Pseudomonas sp. tolerance for hydrocarbon and its initial sensitivity per mortality within 24 hours of exposure could be accessed as indices to measure level of pollution or toxicity of petroleum products.

Bacterial and viral diseases constitute a major factor limiting the successes of shrimp farming industry. In this context, Identification of drug resistant pathogens and determination of the pathogenic processes of shrimp are fundamental for further progress in the disease management. They could be valuable in the evaluation of their epidemiology and control measures. Water samples from the aquaculture environment were contaminated with Vibrio anguillarum, V. damsela, V. furnissii, Aeromonas hydrophila and A. sobria. Both Vibrio spp and Aeromonas spp were shown to be pathogenic and to cause mortality to the infected post larval P. monodon. Vibrio spp. were shown to grow in TCBS agar, zobell’s marine agar and blood agar while Aeromonas spp. additionally exhibited growth in blood agar with selective supplement ampicillin (10 µg/ml) and showed a zone of clearance in DNase agar. LD₅₀ values of V. anguillarim, V. damsela, V. funrissii, A. hydrophila and A. sobria to post larval P. monodon were determined as 1.1 x 10⁵, 2.7 x 10⁵, 6.0 x 10⁵, 8.5 x 10⁴, and 7.5 x 10⁴ bacterial cells/ml respectively. Ciprofloxacin and streptomycin were found to be the most effective antibiotics in the reduction of the mortality of experimentally infected post larvae. The growth of Vibrio spp was inhibited by nifurpirinol (prefuran) at 25 µg/ml, oxytetracycline at 13 µg/ml, erythromycin phosphate at 4 µg/ml, chloramphenicol at 12 µg/ml, streptomycin at 2 µg/ml, and ciprofloxacin at 1 µg/ml. While the growth of Aeromonas spp was inhibited by nifurpirinol (prefuran) at 24 µg/ml, oxytetracycline at 14 µg/ml, erythromycin phosphate at 5 µg/ml, chloramphenicol at 12 µg/ml, streptomycin at 2 µg/ml, and ciprofloxacin at 1 µg/ml. Vibrio spp. was sensitive to o/129 vibriostatic agent (150 µg) whereasAeromonas spp. was resistant to it. The pathogenic bacteria viz. Vibrio spp and Aeromonas spp could be controlled to some extent with judicious use of effective antibiotics to improve the health status of shrimp in culture conditions and can have a positive impact in the areas of sustainable aquaculture environment and development.

A dynamic mediatory role between starch synthesis and degradation has been ascribed to starch phosphorylase. However, plant starch phosphorylase is largely considered to be involved in phosphorolytic degradation of starch. It reversibly catalyzes the transfer of glucosyl units from glucose-1-phosphate to the non-reducing end of glucan chain with the release of inorganic phosphate. It is widely distributed in plant kingdom. Enzyme multiplicity is also common in starch phosphorylase and different multiple forms have been predicted to have different roles in starch metabolism. Here, various biochemical properties have been reviewed. Its regulation by aromatic amino acids has also been discussed. Importance of plastidial and cytoplasmic starch phosphorylase has also been discussed. Various biotechnological aspects have been discussed. Its exploitation in production of glucose-1-phosphate, a cytostatic compound has been discussed in the present review.

Provision of portable drinking water is of public health concern especially in developing countries where this is unavailable. Microbes are ubiquitous and are known to contaminate materials including food and water. Microbial contamination cannot be detected by sight, smell or taste. Basic laboratory tests are the best way to tell if coliform organisms are present as they can be there with no appearance or taste difference. The microbiological quality of drinking water (DW) in Zamfara North Senatorial Zone was examined. A total of 16, two each from each of eight common brands of sachet water in the area were bought from water vendors, and were examined for total bacteria load, total coliform and presence of bacteria species using standard microbiological techniques. The result showed that the total viable count of bacteria in all the samples ranged from 6.0×10² CFU/ml to 4.0×10⁸ CFU/ml. Total coliform was 1.8×10⁷ MPN/100 ml for all the tested samples. The organisms isolated were Pseudomonas maltophila, Escherichia coli, Citrobacter freundii, Pseudomonas pseudomollia, Salmonella typhi, Shigella species, and Pseudomonas dimineta. Prevalence of different isolates revealed that Pseudomonas, C. freundii, S. typhi and E. coli were predominant in comparison to Shigella species. The present study revealed that the microbial quality often exceed World Health Organization (WHO) and Food and Agricultural Organization (FAO) allowable limit of 1.0x10² CFU/ml for potable water and Standards Organization of Nigeria (SON) maximum permissible level of 10 CFU/ml (total coliform) and 0 CFU/100 ml. The high microbial isolates and load may have contaminated the water from the environment. These microbes found in the drinking water sources are known to cause several diseases. Present study indicate that water testing would ensure the supply and availability of contamination-free drinking water; and awareness amongst people towards sanitation and hygienic conditions for storage of drinking water is needed to keep away the use of contaminated water. The present study suggests that drinking water sources should be properly treated prior to consumption using appropriate methods; so as to reduce the occurrence of waterborne diseases.

Host parasite relationships result from prolonged associations between organisms living in a given environment. The nature and extent of the association will determine the type of relationship existing between the co-habiting organisms. Host-parasite associations usually give rise to four main relationships namely parasitism, mutualism, commensalism and phoresis.

Nasal carriers of Staphylococcus aureus are important reservoirs with risk of developing endogenous infections or transmitting infections to susceptible individuals. Methicillin-resistant S. aureus (MRSA) are associated with higher rates of treatment failure. Some strains of S. aureus produce slime which is believed to make the microorganisms more resistant to antibiotics and host defenses. The antibacterial activity of ethyl acetate: n-hexane (EtOAc:HEX) extracts of Mulinum spinosum (5:95% EtOAc:HEX, 50:50% EtOAc:HEX, 70:30% EtOAc:HEX and  mix 20:80/30:70% EtOAc:HEX, 50:50/70:30/100:0% EtOAc:HEX) were assayed against 3 slime-producing S. aureus strains and 2 MRSA strains isolated from nasal carriers. S.aureus ATCC 35556 slime-producing strain and MRSA ATCC 43300 strain were used as controls. The extracts were prepared using flash chromatography. M. spinosum 5:95% AcOEt:HEX showed antibacterial effect against all slime-producing strains (MIC:500 µg/ml) and the highest activity against MRSA strains (MIC:500 to 1000 µg/ml). All M.spinosum extracts assayed were active against slime-producing S. aureus and MRSA at doses between 500 and 4000 µg/ml. Both, slime-producing S. aureus and MRSA are highly contagious and hardly eradicated by antibiotic therapies. So, there is an increasing need to find new substances with the ability to inhibit these strains.

Background: Hepatitis is a liver disease caused by infectious and non-infectious agents. Hepatitis B and C are global health problem. Hepatitis B (HBV) and C virus (HCV) infections are usually occur in the population due to their divers mode of transmission through blood.

Objectives: This study aimed to assess the prevalence of hepatitis B and C virus among population in Babylon province and evaluation of the levels of the TNF-alpha in infected patients with hepatitis B and C.

Methods: The study was conducted on individuals referred to the Central Public Health Laboratory to detected the prevalence of HBV and HCV infections in Babylon province. A total of 6061 samples was involved in this study, they surveyed by VIDAS test for demonstration of both viruses, then confirmed by real- time PCR. Concentration of TNF- α was measured in patients with hepatitis B and C and control group (Control group included healthy individuals with no history of renal or liver diseases and negative results for HBV and HCV test) by using ELISA test.

Results: A total of 6061 individual were examine, amongst them, 18 (0.29%) were positive for HBV and 16 (0.26%) were positive for HCV by both VIDS test and real-time PCR.

Most of HBV infections seen among hemodialysis patients followed by haemophila, while most HCV infections seen among thalasemia and haemophila patients than others groups.

The prevalence of both viruses was higher among male than female, and in urban than rural rejoin. Most of HBV infections were high in individual with age ranging between 20-29 and 30-39 years, while most HCV infections in individual with age ranging between 20-29 years, although it is statistically insignificant.

The TNF- α mean serum level was significantly higher in individual with HBV and HCV infection (68.7±12.84 pg/mL) and (89.1±22.017) as compared with control group (43.1±5.87 pg/mL).

Conclusion: The prevalence of hepatitis B and C virus was relatively low. There are increasing level of TNF-α in the serum of patients with viral hepatitis B and C.

Studies were undertaken on mycotoxin contamination and induced biochemical changes in some important medicinal plants. For the purpose of study four important medicinal plants namely Azadirachta indica, Emblica officinalis, Plantago ovata and Vitex negundo were selected. Samples were collected from different localities of Uttarakhand (India). Mycotoxin producing fungi like A. flavus, A. ochraceus, F. verticillioides and Penicillium citrinum were recorded. In comparison to other mycotoxigenic fungi, percentage toxigenicity was higher in Aspergillus flavus. In case of E. officinalis, 32.69% isolates of A.flavus were toxigenic and produced aflatoxins up to 21 µg/ml in the liquid media followed by A. indica where 22.2% isolates produced aflatoxin in the range of 0.4-13.8 µg/ml. In case of V. negundo and P. ovata 14.28% and 8% isolates of A. flavus elaborated aflatoxin B₁,B₂ and aflatoxin B₁ respectively at low concentration. Mycotoxin contamination in stored samples of E. officinalis and V. negundo showed aflatoxin B₁ and aflatoxin B₁,B₂ as natural contaminant. Twenty eight percent (28%) samples of E. officinalis exhibited higher concentration of aflatoxins up to 0.98 µg/g whereas in case of V. negundo 6% samples were found naturally contaminated with aflatoxin B₁. Alkaloid content of medicinal plant produce understudy was estimated in healthy as well as A. flavus infested samples.There was an indication of inhibition in the total alkaloid content due to the toxigenic strain of A. flavus. Statistical analysis of the results show a decline in the level of total alkaloid content due to fungal contamination significant at 5% level of significance.

Thermophilic Campylobacters are major causes of gastroenteritis in human. The main risk factor of infection is consumption of contaminated or by cross-contaminated poultry meat. In Coteˆ d’Ivoire, gastroenteritis is usually observed but no case of human campylobacteriosis has been formally reported to date. The aims of this study were to determine prevalence and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli isolated from chickens ceaca in commercial slaughter in Abidjan. Between May and November 2009, one hundred and nineteen (119) chicken caeca samples were collected and analyzed by passive filtration method followed by molecular identification (PCR). From these 119 samples, 76 (63.8%) were positive to Campylobacter tests. Among the positive colonies, 51.3% were C. jejuni and 48.7% were C. coli. Of the 39 C. jejuni isolates, 79.5%, 38.5%, 17.9%, 10.3%, and 7.7% were, respectively, resistant, to nalidixic acid, ciprofloxacin, amoxicillin, erythromycin, and gentamicin. Among the 37 isolates of C. coli, 78.4%, 43.2%, 13.5%, 8.1%, and 0% were resistant, respectively, to the same antibiotics. On the other hand, detection of virulence putative gene shows presence of cadF in 100 % of tested strains. In addition, cdtA, cdtB and cdtC genes were detected in 100%; 69.43% and 71.06% respectively of C. jejuni isolates. Moreover, only cdtA gene of cdt genes was detected in 12.82% of C. coli strains tested in this study. In conclusion, we reported in this study the presence of high Campylobacter contamination of the studied chickens. Molecular identification of the bacteria was performed and determination of high resistance to antimicrobials of the fluoroquinolone family was revealed.

Halophilic bacteria are commonly found in natural environments containing significant concentration of NaCl that harbor a number of microbes producing novel bioactive compounds. In this study we aimed to isolate and screen out the halophilic bacteria and to determine their activity for production of the bioactive compounds. Marine soil sediments were collected from the solar saltpans of Thoothukudi District, Tamil Nadu, India. Based on colony morphology, two species were isolated and identification was done by using morphological and biochemical tests. The extracts of cell-free supernatant of the two halophilic isolates were screened for bioactive compound and tested for antimicrobial activity against human pathogenic bacteria such as Staphylococcus aureus, Pseudomonas sp, Klebsiella sp, Vibrio sp, Escherichia coli and fungi Aspergillus niger and Penicillium chrysogenum by the agar cup diffusion method. The results were then compared to standard antibiotics which showed 80% of similar activity in 50 μL/g concentration. In addition, the arbitrary unit of two isolates was calculated against S. aureus which produced enhanced inhibitory results. These findings show that the saline soil of Thoothukudi represents an untapped source of bacterial biodiversity and also that most bacterial isolates are capable of antibacterial and antifungal metabolite production.

Day by day, faith of people on herbal medicine increases due to the side effect of synthetic drugs; this has resulted into people falling back to the traditional knowledge of plant for their health care. Certain local practitioner and traditional healers use the fruits of Citrus aurantium var. Dulcis L. pulps in various disease management and so, they advise to eat the pulps along with the drinking of the juice. The present study deals with preliminary phytochemical analysis of the fruit of Citrus aurantium var. Dulcis L pulp using 95% ethanol for its extraction. The fruits of Citrus aurantium var. Dulcis L. pulp ethanolic extract revealed the presence of all tested phytochemical compounds except protein and glycoside. These include Alkaloids, Tannins, Phenolic, Quinine, Reducing Sugar, Coumarins, Flavonoids, Saponins and Steroids. During the analysis, the quantitative fractionation of the ethanolic extract showed a reasonable amount of saturated hexane fraction (40 g), unsaturated hexane fraction (2.0 g), methanolic fraction (1.3 g), acidic fraction (1.2 g) and basic fraction (0.3 g). These results from the fruit of Citrus aurantium var. Dulcis L. pulps revealed their ignored medicinal importance by throwing it away to domestic animals, contributing to environmental de-sanitation and a natural product waste as medicine. And it’s a needful help for the scientific documentation and standardization of row fruits waste material as to be used in medicine and recommended for worldwide acceptance.

This article describes the use strains thiobacteria Acidithiobacillus ferrooxidans Achi1, Achi3 for decontamination pyrophoric deposits, which are a source of fires in refineries. It is found that the inhibitory factor is the intensity of insolation and organic impurities, and optima conditions for decontamination of hazardous waste are the temperature of 30 ± 35°C, the use of hydrocarbon-oxidizing microorganisms and pH 2.0-2.5.

Vaginal candidiasis (VC) is a common genital tract infection that affects mostly women especially the pregnant women leading to complications as abortions, premature birth, low birth weight among others. Vaginal specimens were collected from 300 pregnant women attending pre-natal visits in three selected hospitals in Nnewi, Anambra State, Nigeria using sterile swab sticks. Specimens were cultured in Sabouraud and blood agar media under sterile condition. All pregnant patients were considered throughout the period of the study, having obtained an informed consent from them. Chi-square analysis of data indicated that ninety patients were positive for candidiasis, a prevalence rate of 30.0%. Furthermore, pregnant women aged 26 to 30 recorded the highest prevalence (13.669; df = 5) that was statistically significant (p<0.05). The women who in their third trimester of pregnancy were mostly infected (6.163; df = 2) with infection status which was highly significant (p<0.05). The symptomatic and asymptomatic patients were uniformly infected with Candida hence, there was no significant difference between them (p>0.05). Further stratification of the pregnant women according to their parity revealed that Candida infection decreased with parity and was significantly higher in those in their first and second pregnancy (p<0.01). Consideration of the socio-economic status and occupation of the pregnant women showed no significant difference (p>0.05).  It was suggested that avoidance of predisposing factors, Vaginal Candidosis could resolve within a short period of time.

Aims: To determine the carrier rate of typhoid and paratyphoid organisms in Enugu community.

Place and Duration of Study: The project was carried out in Enugu Urban in South Eastern part of Nigeria. Those enrolled in the study that lasted for one year are students, traders and civil servants. 

Methodology: Ninety six apparently healthy adults without any complaints, comprising of forty eight males and forty eight females were selected for the study. Blood, urine and stool samples were collected from all in the study group. The samples were subjected to standard bacteriological culture for enteric organisms at the Medical Microbiology Laboratory of University of Nigeria Teaching Hospital, Enugu.

Results: Salmonella typhi was isolated from the stool of two (2.08%) of the candidates while S. paratyphi was recovered from the stool of another individual (1.04%) of the study group. No candidate gave Widal titre greater than 40 to ^,O^, agglutinin  which is statistically significant (Anova F value 597.7, P value <0.0001). Only Two Candidates positive for S. typhi culture gave titre of 160 to^, H^, antigen, also statistically significant (Anova F value 1195, P value <0.0001).

Conclusion: Typhoid carrier rate in Enugu Community is 2.1% while that of paratyphoid is 1.0%. From the study females are more likely to be typhoid carriers in Enugu.  

A survey was conducted between May and July to assess the extent of loss in mangoes at wholesale and consumer levels caused by fungal spoilage during post-harvest. Mango fruits were purchased from different markets in Saudi Arabia and the degree of losses due to fungal spoilage was assessed at the different levels of marketing. Fungal spoilage was found to be the highest at the consumer level and least at the wholesale level. Aspergillus flavus rot, Aspergillus niger rot, Fusarium sp. and Penicillium spp. rot were the commonest diseases affecting the mango fruits.