Advanced Journal of Microbiology Research

Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (1), pp. 001-012, January, 2019. © International Scholars Journals

Full Length Research Paper

The effect of soil water potential on survival of fecal coliforms in soil treated with organic wastes under laboratory conditions

Ali Akbar Safari Sinegani* and Javad Maghsoudi

Faculty of Agriculture, Bu-Ali Sina University, Hamedan, Iran.

Accepted 25 January, 2019

Abstract

Pollution caused by animal wastes has become a great problem in many countries. The objective of this study was to test the effects of water potential on survival of fecal coliforms in a soil treated with 3 manures. A semiarid soil was treated with raw cow and poultry manures (CM and PM) and sewage sludge (SS) at a rate of 20 g kg^-1 (dry weight basis). Three water potentials established for soil incubation were: Saturation (SAT, 0 bars), field capacity (FC, -0.3 bars), and permanent wilting point (PWP, - 15 bars) . Fourth irrigation treatment was drying-rewetting cycle (DWC) between -0.3 to -15 bars. Colony forming units of Escherichia coli and fecal coliforms on EMBA (eosin methylene blue agar) were counted during soil incubation. The population of E. coli was higher in the soils treated with CM but the populations of lactose positive and negative coliforms were higher in the soils treated with PM. The populations of E. coli and other fecal coliforms were significantly higher in the soils incubated in SAT compared to those in soils incubated in other water potentials especially in the early stages of soil incubation. The populations of fecal coliforms were decreased significantly with increasing time of incubation. Survival of E. coli were near 40 days in the soils treated with PM and SS. E. coli could survive in the soils treated with CM and incubated in SAT and PWP for more than 90 days may be due to low level of negative interactions in these unsuitable water conditions.

Key words: Organic wastes, soil water potential, Escherichia coli, fecal coliforms, survival, incubation.

Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (1), pp. 001-006, January, 2019. © International Scholars Journals

Full Length Research Paper

A novel vector for expression cloning of large numbers of polymerase chain reaction products in Pichia pastoris

Dongming Lan¹, Wenkai Wang¹, Lijuan Sun¹, Yonghua Wang²* and Bo Yang¹

¹School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510641, China.

²College of Light Industry and Food Sciences, Key Lab of Fermentation and Enzyme Engineering, South China University of Technology, Guangzhou 510641, China.

Accepted 8 December, 2018

Abstract

We described construction of a novel vector, pAOX-HT, for direct cloning of polymerase chain reaction (PCR) amplified fragments and expression in Pichia pastoris. The pAOX-HT serves both as a T-vector and expression vector and can be generated from a parent plasmid by digestion with restriction enzyme XcmI. To minimize the non-recombinant background, the parent vector pAOX-HT-BSK was designed to contain a large insert. The cloning efficiency was above 95% when tested with PCR products. The linearized pAOX-HT was engineered to harbor a potential AflII site (CT) upstream of the T/A cloning site. An AflII site was reconstructed when the PCR product with 5’-TAAG sequence was ligated into the T/A cloning site. Taking advantage of this property, we digested the ligated products by restriction enzyme AflII before transformation to eliminate the clones containing inserts with undesired orientation. By using pAOX-HT vector, the lipase B gene from Candida Antarctica was efficiently cloned and expressed in P. pastoris and the recombinant protein was purified by affinity chromatography. These results demonstrate that the pAOX-HT might serve as a useful tool for gene function study.

Key words: T vector, expression vector, yeast expression, restriction endonuclease digestion.

Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (1), pp. 001-006, January, 2019. © International Scholars Journals

Full Length Research Paper

Identification and quantitation of conjunctival aerobic bacterial flora from healthy residents at different ages in Southwest China

Jing Liu, Jing Li, Jian Huo and Hanping Xie*

Southwest Hospital, Southwest Eye Hospital, Third Military Medical University, Chongqing, China 400038, China.

Accepted 20 January, 2019

Abstract

To identify and quantitate conjunctival aerobic bacterial flora from healthy residents of three age groups in Chongqing city, Southwest China. Samples taken with moist swabs from the lower fornix of bilateral eyes of 33 children (9.85 ± 0.57 years old), 31 youths (21.23 ± 0.88 years old), and 30 elders (72.97 ± 4.94 years old) respectively were tested for bacterial cultures. Positive cultures were identified and colony forming units (CFUs) were counted. Bacterial quantity was expressed as the number of CFUs divided by the weight difference of each sample transport swab-tube measured before and after the sample collection. The elders showed a higher isolation frequency (93.33%) of conjunctival aerobic bacteria than that of children (45.45%) and youths (50.00%), (P<0.01). Corynebacterium diphtheroid, Staphylococcus epidermidis and Staphylococcus aureus isolated from the elders all had a greater percentage. The elders also had a larger number of aerobic bacteria species per eye than did children and youths (P<0.01). Children showed a lower quantity of conjunctival aerobic bacterial flora than that of the other two groups (P<0.01). C. diphtheroid was the most numerous isolate from all the subjects. The quantity of C. diphtheroid became larger when residents grew older. S. epidermidis was the second most numerous bacteria in the groups of children and elders, and the elder group had a greater number of S. epidermidis than the children group (P<0.01). The number of species and quantity of conjunctival aerobic bacterial flora from the elders were larger than those from children and youths, which should be of concern to an ophthalmologist.

Key words: Conjunctival sac, aerobic bacteria, age, quantitative analysis, microecology.

Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (1), pp. 001-007, January, 2019. © International Scholars Journals

Full Length Research Paper

A method for the determination of tannase activity based on gallic acid measurement by high performance liquid chromatography (HPLC)

Nalan Yilmaz Sariözlü¹*, Erdoğan Çakir¹, Merih Kivanç¹ and Muzaffer Tunçel²

¹Department of Biology, Faculty of Science, Anadolu University, 26470 Eskişehir, Turkey.

²Department of Analytical Chemistry, Faculty of Pharmacy, Anadolu University, 26470 Eskişehir, Turkey.

Accepted 11 December, 2018

Abstract

A new high performance liquid chromatography (HPLC) method was developed based on an indirect determination of tannase activity through gallic acid measurement using paracetamol as an internal standard. The separation was performed on a C₁₈ column using a mobile phase consisting of aqueous formic acid solution (1%) and methanol (85:15; v/v) pumped at 1 ml min^-1. Samples (10 μl) were injected and signals were detected at 254 nm. Repeatability was 0.81 and 1.28 for peak normalization values of gallic acid (4.16 x 10^-5 M) and internal standard (2.02 x 10^-5 M). A good linearity was shown in the range of 1.04 x 10^-5 to 8.32 x 10^-5 M. Limit of detection (LOD) and limit of quantification (LOQ) values were calculated to be 2.2 x 10^-6 M and 6.6 x 10^-6 M, respectively. The applicability of the method was tested using Penicillium spinulosum which produces tannase. The method was highly sensitive, accurate, reliable, and repeatable and at the same time it is applicable to both pure and crude enzyme.

Key words: Tannase activity, gallic acid, tannic acid, high performance liquid chromatography, enzyme activity, validation.

Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (1), pp. 001-005, January, 2019. © International Scholars Journals

Full Length Research Paper

Antibacterial activity of Moringa oleifera and Moringa stenopetala methanol and n-hexane seed extracts on bacteria implicated in water borne diseases

Atieno Walter^1*, Wagai Samuel¹, Arama Peter¹ and Ogur Joseph²

¹Department of Botany and Horticulture, Maseno University, Kenya.

²Department of Chemistry, Maseno University, Kenya.

Accepted 11 November, 2018

Abstract

The anti-bacterial activity of methanol and n-hexane extracts of Moringa oleifera and Moringa stenopetala seeds was conducted on 3 bacterial species (Salmonella typhii, Vibrio cholerae and Escherichia coli) which normally cause water borne diseases. The paper disc diffusion method was used with treatments arranged in a completely randomized design and replicated four times. The highest inhibitions were observed at dilutions of 20, 5 and 40% for M. oleifera and M. stenopetala methanol extracts on E. coli, S. typhi and V. cholerae respectively. The n-hexane extract of both M. oleifera and M. stenopetala had a higher inhibition on S. typhii than V. cholerae and E. coli. The results of this study showed that M. oleifera and M. stenopetala had a degree of antibacterial properties especially in low doses.

Key words: Moringa oleifera, Moringa stenopetala, methanol and n-hexane extracts, inhibition, Salmonella typhii, Vibrio cholera, Escherichia coli, Kenya.

Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (1), pp. 001-004, January, 2019. © International Scholars Journals

Full Length Research Paper

Some cellular immune components and C-reactive protein monitoring in female handball players during a competitive period

Tarik Sevindi¹, Serkan Hazar¹, Serkan Ibi ¹ and Kadir Gökdemir²

¹Nigde University, Physical Education and Sports Department, Nigde, Turkey.

²Gazi University Physical Education and Sports Department, Turkey.

Accepted 6 January, 2019

Abstract

The aim of the study was to determine the effect of the competitive period training on leukocytes, some leukocyte subgroups and C -reactive protein (CRP) in sportswomen playing in the professional handball league. The average age of the group was 20.6 ± 3.37 years, height average was 163 ± 6.30 cm, weight average was 58.73 ± 4.92 kg. Blood samples were taken before and after the training period of the competition. Subjects rested for 48 h until taking blood samples. Subsequently, leukocytes, some leukocyte subgroups and CRP were determined. Leukocytes and leukocyte subgroups were determined using a Roche Sismex 2000XL device. CRP was determined using Beckman immunochemistry systems in-vitro diagnostic kits and nephelometric methods (Beckman Coulter Array 360 System, USA). Statistical analysis was performed using a paired t-test. p values <0.05 were considered statistically significant. In this study no significant difference was found on WBC and granulocyte values between pre and post training period. However, significant differences in lymphocyte values were measured. After training period, lymphocytes declined (p<0.05), but no significant differences in monocyte levels were determined. CRP values after the training period were significantly decreased (p<0.05), as compared to pre training values. We can conclude that competition period training did not significantly affect the cellular immune elements, but caused a significant decrease on CRP values, which are markers in chronic inflammation.

Key words: Training, immune component, C- reactive protein, female athlete.