Advanced Journal of Microbiology Research

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 19 (1), pp. 001-006, January, 2025. https://internationalscholarsjournals.org/© International Scholars Journals

Full Length Research Paper

Optimizing Blastocyst Formation from Cryopreserved Mouse Two-Cell Embryos: The Role of Fibroblast and Hepatocyte Growth Factors

Fatemeh Ghasemian¹, Mahnaz Azarnia², Abtin Heidarzadeh³, Shervin Ghadarjani⁴ and Mohammad Hadi Bahadori⁵*

¹Department of Biology, Faculty of Science, Kharazmi (Tarbiat moallem) University, Tehran, Iran.
²Department of Biology, Faculty of Science, Kharazmi (Tarbiat moallem) University, Tehran, Iran.
³Department of Community Medicine, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran.
⁴Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran.
⁵Cellular and Molecular Research Center, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran.

Accepted 5 September, 2022

Abstract

There is great need to improve our understanding of what increases an embryo’s development potential, after vitrification-thawing processes. For this subject, 358 two-cell stage embryos were collected from oviduct of pregnant two-day old mice and vitrified. After thawing, embryos were cultured in Tyrode's (T6) medium supplemented with different doses of fibroblast growth factor (FGF; 0, 10, 20, 50 and 100 ng/ml) and hepatocyte growth factor (HGF; 0, 10, 20, 50 and 100 ng/ml) until the blastocyst stage. To determine quality of blastocysts, blastocysts were stained with hoechst and propidium iodide. After culture for 24 h, 92.65% of treated embryos with 20 ng/ml of FGF had higher (P<0.05) survival rate in comparison to the control group (84%). Blastocyst embryo formation rate were 79.43% (P< 0.01), and 67.46% (P< 0.05) in the treated groups with 20 ng/ml of FGF and 20 ng/ml of HGF, respectively, which were significantly different from the control (56%). In the treatment group with 20 ng/ml of FGF, blastocysts with >64 cells had a significantly higher inner cell mass (ICM) in comparison to the control group (P< 0.01). In conclusion, in this experiment, addition of growth factors in the culture had favorable effects on post-thawed cleavage of vitrified 2-cell embryos and blastocyst quality.

Keywords: Blastocyst quality, growth factors, preimplantation development, vitrification.
 

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 18 (8), pp. 001-004, August, 2024. Available online at www.internationalscholarsjournals.org © International Scholars Journals

Full Length Research Paper

Direct Detection of Salmonella InvA, SpiC, and SipC in Clinical Samples Using Dot Blot Hybridization

Hang’ombe Bernard Mudenda¹*, Ulaya William¹, Mwansa James C. L.², Mubita Charles¹, Isogai Nayuta³, Mulenga Evans¹, Moonga Ladslav¹, Isogai Hiroshi⁴ and Isogai Emiko⁵

¹School of Veterinary Medicine, University of Zambia, P. O. Box 32379, Lusaka, Zambia.
²University Teaching Hospital, Department of Pathology and Microbiology, Lusaka, Zambia.
³Department of Agro-bioscience, Faculty of Agriculture, Iwate University, Morioka 020-8550, Japan.
⁴Animal Research Center, Sapporo Medical University, 060-8556, South 1 West 17, Chuoh-ku, Sapporo, Japan.
⁵Laboratory of Animal Microbiology, Department of Microbial Biotechnology, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori Amamiya-Machi Aoba-ku, Sendai 981-8555, Japan.

Accepted 28 March, 2024

Abstract

Pathogenesis of Salmonella depends upon a large number of factors controlled by an array of genes that synergise into actual virulence. The goal of this study was to detect Salmonella invA, spiC and sipC directly from clinical specimens, using the dot blot hybridization assay. We detected invA , spiC and sipC as a one combination from 4.5% (95% CI: 2.21 to 8.64) human feacal and 35.2% (95% CI: 26.4 to 45.0) poultry samples after enrichment. Furthermore the dot blot method had a higher sensitivity than routine culture, before and after enrichment. These results indicate that dot blot hybridization may be used to directly detect Salmonella invA, spiC and sipC in clinical samples.

Key words: Salmonella, dot-blot hybridization, spiC, sipC, invA, clinical-samples.
 

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 18 (8), pp. 001-005, August, 2024. Available online at www.internationalscholarsjournals.org © International Scholars Journals

Full Length Research Paper

Genetic Variability of Helicobacter pylori in Iran: Clarithromycin Resistance and 23S rRNA Mutations

Mohammad Kargar¹*, Maryam Baghernejad¹, Abbas Doosti² and Sadegh Ghorbani-Dalini^1,3

¹Department 1Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran.
²Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran.
³Jahrom Branch, Young Researcher's Club, Islamic Azad University, Jahrom, Iran.

Accepted 12 April, 2024

Abstract

To determine the 23S rRNA point mutations in clarithromycin resistance of Helicobacter pylori strains isolated from southwest, Iran. This was a cross-sectional survey, which was done on 263 patients who referred to endoscopy department of Shehrekord university of medical sciences. According to gram stain, urease, catalase, oxidase and polymerase chain reaction (PCR) H. pylori identified. Standard National Committee for Clinical Laboratory Standard (NCCLS) method used for assessment of clarithromycin resistance. Specific primers and restriction enzymes BsaI and MboII by PCR-RFLP were used for analysis of A2143G and A2142G mutations. So for the detection of A2142C, specific primers and PCR method were used. 84 strains of H. pylori (31.94%) determined by PCR method. Of 19 (22.62%) clarithromycin resistant strains 13 (68.40%), 3 (15.78%), 2 (10.52%) had A2143G, A2142G, A2142C respectively and one unknown mutation in 23S rRNA gene. Because of considerable resistance to clarithromycin, direct diagnosis of this mutation by molecular approach in other parts of the country is necessary.

Key words: Polymerase chain reaction, clarithromycin, resistance, 23S rRNA, polymerase chain reaction - restriction fragment length polymorphism.
 

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 18 (7), pp. 001-005, July, 2024. Available online at www.internationalscholarsjournals.org © International Scholars Journals

Full Length Research Paper

Assessing Genetic Relationships and Population Differentiation in Karakul Sheep Using Microsatellite Markers

Shahram Nanekarani¹*, Cyrus Amirinia² and Nour Amirmozafari³

¹Department of Animal Science, Faculty of Agriculture, Broujerd Branch, Islamic Azad University, Broujerd, Iran.
²Department of Animal Biotechnology, Animal Science Research Institute of Iran, Karaj, Iran.
³Department of Molecular Biology, Iran University of Medical Sciences, Tehran, Iran.

Accepted 22 March, 2024

Abstract

In this study, the genetic variation in Karakul sheep was investigated using 15 microsatellite markers (MCMA2, BMS460, BM1815, OARCP26, OARFCB20, MAF64, OARAE129, BMS332, LSCV38, BM6444, BMS995, MCMA26, BMS678 and OARCP49) and all fifteen loci were amplified successfully. Genomic DNA was extracted from 120 blood samples, using modified salting-out method. Tests of genotype frequencies for deviation from the Hardy-Weinberg equilibrium (HWE) were performed at each locus and revealed significant departure from HWE (P < 0.001) due to heterozygote excess. Parameters of variability such as effective number of alleles and gene diversities corroborated with the high level of variation frequently displayed by microsatellite markers. The fifteen tested loci were all polymorphic. Furthermore, other criteria of genetic variation including polymorphism information content (PIC) values and Shanon information index were calculated in this study. Results showed that, high level of genetic diversity was observed in Karakul breed and this breed was not at risk for conservation concept. This research has also shown that microsatellite technique is a useful tool for evaluation of genetic variation among domesticated animals.

Key words: Karakul sheep, microsatellite markers, genetic variation, polymorphism. Hardy-Weinberg equilibrium (HWE).

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 18 (7), pp. 001-006, July, 2024. Available online at www.internationalscholarsjournals.org © International Scholars Journals

Full Length Research Paper

Rapid and Accurate Detection of Agrobacterium tumefaciens in Soil Using PCR

Wei Yang^1,2, Lei Ji³, Li-Rong Tan², Shi-Mo Li¹, Yan Wang³, Hong-Xia Liu²*, and Yu-Ming Luo¹*

¹Jiangsu Key Laboratory of Eco-Agricultural Biotechnology around Hongze Lake, Huaiyin Teacher’s College, No. 111, Changjiang West, Huai’an, 223001, Jiangsu Province, China. ²Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China. ³Shanghai Forestry Station, Shanghai, 200072, China.

Accepted 28 March, 2024

Abstract

One pair of primers was designed based on the sequence of tmr locus for specific and sensitive detection of Agrobacterium tumefaciens. Only the A. tumefaciens strain can produce the 236bp target fragment among the fourteen bacterial species that tested. The sensitivity of the specific PCR system was determined by a nested-PCR amplification which can numbered the copies of the template DNA. According to the results, it can give positive band when only 100 copies were in the template. The protocol was carried out for detection A. tumefaciens of twelve soil samples collected from six different gardens in Shanghai where crown gall happened. Two of the samples which collected from symptomless gardens also give the positive band. Based on the results we can make a conclusion that this pair of primers can be a useful tool in detecting A. tumefaciens, especially in detecting latent infection of this devastating pathogen.

Key words: Agrobacterium tumefaciens, detection, polymerase chain reaction (PCR).
 

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 18 (7), pp. 001-013, July, 2024. Available online at www.internationalscholarsjournals.org © International Scholars Journals

Full Length Research Paper

Harnessing Wastewater Microbes for Polyhydroxybutyrate Production: A Case Study with Enterobacter aerogenes 12Bi

Nur Ceyhan¹* and Guven Ozdemir²

¹Department of Biology, Faculty of Sciences, Mugla University, Mugla-48170, Turkey.

²Basic and Industrial Microbiology Section, Department of Biology, Faculty of Sciences, Ege University, Izmir-35100, Turkey.

Accepted 1 March, 2024

Abstract

A strain  of  poly- -hydroxybutyrate (PHB)-accumulating  bacterium  was isolated  and  identified  as Enterobacter aerogenes (designated E. aerogenes 12Bi) by using biochemical and phylogenetic characterization. The accumulation of a large amount of granules in its cells cultured in the domestic wastewater medium (DWWM) were showed by transmission electron microscopy (TEM). When PHB production by our strain was determined by Hypochlorite method, it was found that PHB production ranged from 16.66 to 96.25% (w/w). The highest PHB yield by our microorganism was up to 96.25% within 18 h in DWWM 5 (supplemented with 100% DWW). This is the first report of the use of DWW for production of PHB by E. aerogenes. The results obtained in the study demonstrated that PHB could be efficiently produced to a high concentration with high productivity by using DWWM as an inexpensive substrate. Thus, it can contribute to the reduction of high production cost of PHB.

Key words: Poly- -hydroxybutyrate (PHB), bioplastics, domestic wastewater, Enterobacter aerogenes, biochemical identification, phylogenetic identification.