Open Access Original Research Article

Tsetse and Other Biting Flies in Five Districts of Kaura Local Government Area, Kaduna State, Nigeria

A. J. Dadah, E. I. Ikeh, G. Ayanbimpe, S. O. Omotainse, P. M. Dede, A. C. Igweh

Journal of Advances in Microbiology, Page 1-5
DOI: 10.9734/JAMB/2017/35455

A survey of tsetse and other biting flies in Kaura LGA of Kaduna State was under taken. The study was undertaken to determine the presence of tsetse flies, species, their infection rate, distribution and other biting flies in the study area. The study was carried out between September and November, 2010. A total of six sampling sites were considered. The same number of traps was deployed in each sampling site during the study period. Twenty biconical and NITSE traps were used for trapping tsetse flies. A total of 104 biting flies were caught. These were made up of 32 tsetse flies, 55 Tabanus, 14 Stomoxys and 3 Chrysops species. Out of the 32 tsetse flies, 28 were identified to be Glossina palpalis (16 females and 12 males), while 4 were G. tachinoides (3 females and 1 male). The result of fly dissection showed 3 (9.4%) flies were infected with trypanosomes. Out of this number, 2(6.3%) were female G. p. palpalis and 1(3.2%) was a female G. tachinoides, all infected with T. vivax. The implication of this finding is that the preponderance of tsetse flies and other haematophagous flies, with potentials as mechanical transmitters of T. vivax, could indicate a high likelihood of trypanosomiasis in the study area.


Open Access Original Research Article

Effects of Processing Techniques on the Microbial Quality, Nutritional and Mineral Composition of Black-eyed Bean (Vigna unguiculata)

M. P. Uko, M. D. Umoren, M. P. Bassey, S. I. Umana, O. D. Akan

Journal of Advances in Microbiology, Page 1-6
DOI: 10.9734/JAMB/2017/36554

Introduction: Black eye pea (Vigna unguiculata), because of its preferred taste and nutritional value is one of the commonly eaten beans in Nigeria.

Aim: The effect of processing techniques like soaking, dehulling and drying on microbial quality, nutritional and mineral composition of black-eyed bean was studied.

Study Design: One cup of black-eyed beans was divided into two portions. One portion was manually selected, soaked, dehulled and sundried. The other portion received no treatment. Both portions were ground and sieved.

Methodology: One gram of each sieved sample was serially diluted and enumerated of bacteria using standard microbiological procedures. Proximate and mineral compositions were determined by standard analytical procedures.

Results: The presence of Staphylococcus, Klebsiella, Bacillus, Enterobacter and Salmonella species were observed in both samples. Species of Lactobacillus and E. coli were found in treated and untreated sample, respectively. The protein, ash and moisture contents of the treated sample were higher compared to the untreated sample while that of carbohydrate, fibre and fat were lower in the treated sample. Analysis of zinc and iron revealed higher values in the treated sample than in the untreated sample. In contrast, the concentration of magnesium was slightly lower in the treated sample.

Conclusion: Processing of raw food items such as beans by soaking, dehulling, drying, etc. before cooking confer quality, nutritional and health benefits on the final edible food products.


Open Access Original Research Article

Activities of Locally Formulated and Commercial Effective Microorganisms in Composting of Organic Solid Wastes

G. G. Ezeagu, U. J. J Ijah, O. P. Abioye, B. E. N. Dauda

Journal of Advances in Microbiology, Page 1-15
DOI: 10.9734/JAMB/2017/36292

This study was carried out to evaluate the effectiveness of locally formulated Effective Microorganisms (EM) in the degradation of organic solid wastes. The study was laid out in a Completely Randomized Block Design with three replicates each. The research was conducted at the Centre for Genetic Engineering and Biotechnology, Federal University of Technology, Minna, Nigeria between May 2015 and May 2016. Microorganisms were isolated from waste dumps and were identified as bacteria (Lactobacillus plantarum, Streptomyces griseus, Streptomyces rochei, Bacillus subtilis, Rhodopseudomonas palustris), moulds (Aspergillus oryzae, Aspergillus niger) and yeast (Saccharomyces cerevisiae). The microorganisms were used to formulate Effective microorganisms (EM-A and EM-B), which was applied for biodegradation of organic solid wastes in compost. The activities of the two EM brands were compared with that of commercial effective microorganisms (EM-C) using physicochemical and microbiological parameters during the composting process. The compost treated with EM-A attained the highest temperature of 58ºC. The pH of all the treatments was alkaline at the end of composting, while moisture contents were below 50%. The total aerobic heterotrophic bacterial counts (TAHBC) showed that the highest count of 3.0 × 107cfu/g was recorded in P1, Organic waste + EM-A on the 15th day of composting. The fungal counts varied among the treatments with the highest counts of 1.4 × 104 cfu/g recorded in P3, Organic waste + Commercial EM (EM-C) on the 15th day. Comparing the fertilizer value of compost obtained from the different treatments using their chemical properties, OF1 (Organic fertilizers with EM-A), had the best fertilizer value based on the N-P-K values (2.48 g/kg, 2.48 mg/kg and 2.51 cmol/kg respectively) and therefore, is most suitable for crop use. The activities of Effective microorganisms were responsible for the significant differences in physicochemical and microbiological parameters.


Open Access Original Research Article

Microbiological Quality of Raw, Boiled and Fermented Breadnut Seed (Artocarpus camansi) -Used as Condiment

A. Abimbola Noah, O. Oluwafemi Ogunfowote

Journal of Advances in Microbiology, Page 1-9
DOI: 10.9734/JAMB/2017/36577

Aims: The aim of the study was to investigate the microbial safety and quality assessment of processing breadnut (Artocarpus camansi) into boiled and fermented product for 72 hours at room temperature (30°C±2°C) as condiment.

Methodology: The raw seed was boiled with 0.07% salt as the boiled sample and fermented breadnut seeds were washed, boiled (2hrs), dehulled and wrapped in blanched plantain leaf. It was later boiled again for 2 hours, drained, cooled and allowed to ferment naturally for 72hrs while the raw sample serve as the control. The microbiological quality of the three samples was determined. Isolates were further characterized and identified base on cultural and biochemical characteristics.

Results: The mean total viable bacterial count of raw, boiled and fermented sample decreased from 4.22±0.31 to 1.04±0.16log10cfu/g. Staphylococcus count range from 3.21±0.13 to 1.04±0.07 log10cfu/g. Coliform count range from 2.31±0.43 to 1.13±1.06 log10cfu/g. Fungi count range from 2.24±0.07 to 1.02±0.16 log10cfu/g, respectively while there was no growth of Salmonella in the samples. There were significant differences in all the attribute rated for the three samples at ( ). Bacteria isolates were identified as Bacillus subtilis, Bacillus laterosporus, Bacillus coagulans, Bacillus licheniformis, Bacillus pumilus, Micrococcus luteus, Micrococcus varians, Corynebacterium sp, Enterobacter cloacae, Escherichia coli, Staphylococcus aureus and Staphylococcus sp. Fungi isolates were identified as Rhizopus nigricans and Saccharomyces cerevisiae. The predominant microbes in the samples are Bacillus subtilis and Saccharomyces cerevisiae which are beneficial microbes. The coliform, Micrococcus and Staphylococcus present in the 0 and 24hrs fermented seeds were completely eliminated in the 72 hrs fermented sample.

Conclusion: This research has proven that fermented breadnut seed can be used as condiment in soup because it consist of  beneficial microorganisms which increases the nutritional value of food and helps to reduce food-borne diseases microbes that can be hazardous to health.


Open Access Original Research Article

An Effective Cloning, Heterologous Expression and Physiological Activity in Lactococcus lactis NZ9000 of Catalase Gene from Escherichia coli DH5α

Chongbi Li, Sainan Li, Chen Li, Yansheng Liu, Gao Feiyun

Journal of Advances in Microbiology, Page 1-16
DOI: 10.9734/JAMB/2017/34608

In order to clone active Catalase gene from bacteria, we introduced a method of shotgun integrating specific screen from Escherichia coli DH 5a. Genome DNA was extracted from E. coli DH5 and partially digested with Sau3AI. Then some fragments more than 2.26kb were collected and ligated with T4 DNA ligase into the BamHI-cleaved plasmid pUC18 and transformed competent E. coli TG1 cells. The transforments were incubated anaerobically on brain heart infusion (BHI) containing tannic acid. The method could detect catalase activity and screen the catalase-positive clones. The catalase protein can be identified by SDS-PAGE. The results showed that the recombinant plasmid pUC18-kat was constructed successfully by PCR identification and restriction enzyme digestion. Opening the reading frame from DNA Sequence DH5a katE, the sequence length is 2262nt and coding protein are 753 Amino Acids, Molecular Weight is 84198.72 Daltons. This method is simple, and possess of popularization and application value. It would lay a foundation for cloning an active catalase later. Further, in this study, the fragment of 2,262 bp catalase gene katE was cloned into the expression vector pQE30 and transformed into Escherichia coli M15, and KatE protein was expressed after the induction with Isopropylthio-β-D-galactoside. The KatE protein was separated on SDS-PAGE and recovered using a His-tag affinity. New Zealand white rabbits were immunized with the purified protein to harvest polyclonal antibodies. Since L. lactis has no catalase, katE was inserted into Escherichia coli - L. lactis shuttle vector pMG36e and electro-transformed into L. lactis NZ9000. The expression of the KatE protein was confirmed by SDS-PAGE analysis and Western blot. Further experiment demonstrated that the expression of the KatE gene in L. lactis NZ9000 is able to produce active catalase that can provide efficient antioxidant activity. Additionally, to understand the import of catalase katE gene of Lactococcus lactis on the body’s physiological changes of immune function in mice, Enzyme-linked immunoassay (ELISA) was used on the blood of mice, to measure IgG, IgE, CD4 and CD8 levels, and find out whether there is a difference between Mice Groups eating the recombinant L. lactis NZ9000 and other groups i.e. (recombinant E. coli DH5α and L. lactis NZ9000; E. coli DH5α and saline). The experiments showed that recombinant L. lactis NZ9000 was significantly higher than the other on IgG concentrations; IgE, CD4 or CD8 levels are not significant. Mice with increased IgG levels of IgG after the intake of recombinant L. lactis NZ9000 helps to explain the recombinant L. lactis NZ9000 regulatory role in humoral immunity of mice; IgE level did not change, CD4 and CD8 levels were also not changed, thus suggesting that there is no significant effect of cellular immunity on the body in a short time.