MOSP - Macao Scientific Publishers

Research Article

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

Fatemeh Ghasemian, Mahnaz Azarnia, Abtin Heidarzadeh, Shervin Ghadarjani and Mohammad Hadi Bahadori*

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 (P64 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. [...] Read More.

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

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Research Article

Evaluating Growth Regulator Effects on Two Differently Cultured Explants of Carapa guianensis in vitro

Larisse Lobo de Oliveira, Anaíze Borges Henriques, Andrea Furtado Macedo,*

Abstract: Carapa guianensis Aubl. (Meliaceae), known locally as andiroba, is a multi-use species from Amazonia. Andiroba oil is considered an important natural product in the Brazilian market, and international demand is increasing due to its cosmetic and pharmaceutical potential. C. guianensis trees produce seed irregularly over different harvest periods, leading to inconsistent oil production and difficulties with supply. No management plans or protocols have been developed for in vitro or clonal production of Carapa seedlings and the maintenance of genetic resources. The objective of this study was to assess the effect of growth regulators on explants (young leaves, old leaves and apical buds). Explants consisting of leaf segments 1 cm on a side were cultivated in MS medium with and without growth regulators. Evaluation was based on fresh and dry weight of the explants after 20 days. In the media with 2,4-dichlorophenoxyacetic acid (5, 15, 35 or 45 µM), changes were observed in weight and explant appearance (callus). Bud breakage and development of shoots were achieved using 5 µM of 6-benzylaminopurine. Overall, the results showed that 2,4-dichlorophenoxyacetic acid stimulates callus formation on andiroba foliar explants, while 6-benzylaminopurine was superior to thidiazuron for the initial development of shoots.[...] Read More.

Keywords: Growth regulators, Carapa guianensis, in vitro, tissue culture, organogenesis.

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Research Article

Exploring Callus Proliferation and Somatic Embryogenesis in Gossypium hirsutum L.

Ikram-ul-Haq

Abstract: Somatic embryogenesis and plant regeneration are fundamental to tissue culture biotechnology in cotton (Gossypium hirsutum L.) cv. Coker 312. Callus proliferation was considered best on MS1a (2.0 mg/L NAA; 0.1 mg/L ZT; 0.1 mg/L KT) when 6 weeks old callus was cultured from MS1b (0.1 mg/L 2, 4- D; 0.5 mg/L KT) medium, there is no need to select embryogenic calli for somatic embryogenesis, as all of them were converted to somatic embryos. NH4NO3 play an important role in differentiation of callus into somatic embryos but is lethal for embryos just after two weeks. However, KNO 3 is less efficient for somatic embryo induction but is best for embryo maturation. By this procedure 56.51% cotyledenary embryos were developed within 5 weeks. Of that, 82.05% cotyledenary embryos were developed not only into normal plantlets, but rooted simultaneously when cultured on MS (with 0.05 mg/L GA3) medium. A complete plant of Cocker-312 could be regenerated through somatic embryogenesis within 4 to 5 months.[...] Read More.

Keywords: Gossypium hirsutum L,plant regeneration, Coker 312, callus induction, somatic embryogenesis, in vitro regeneration.

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Research Article

In Vitro Regeneration of Hybrid Cashew Plantlets (Anacardium occidentale L.) via Embryo Culture Techniques

Aliyu, O. M.* and Awopetu J.A.

Abstract: Embryos from immature nuts of cashew (Anacardium occidentale L.) were cultured in vitro to regenerate improved hybrid plantlets. Explants (embryo) were excised from developing F1 hybrid immature nuts derived from diallel cross and harvested at 2-, 4-, 6- and 8-weeks after pollination (WAPo) for in vitro culture. The explants were surface sterilized, aseptically dissected and cultured into pure basal Murashige and Skoog (MS) agar medium and MS medium supplemented with 1 mM each of naphthaleneacetic acid (NAA), benzyladenine (BA) and gibberellic acid (GA3) and subsequently observed for germination and survival rates until successful ones were transferred to the field. Age of explants was found to significantly influence both the germination and survival rates. Explants of 6 weeks old and above were found to give better germination rate and highest survival percentage in this study. Only MS medium supplemented with 1 mM of gibberellic acid (MS+GA3) supported germination and growth at 2-WAPo, suggesting the essentiality of GA3 as a growth regulator to a very young cashew embryo. Analysis also showed that factors such as medium composition, age of embryo and genotype (accession) significantly influence the germination rate of cashew embryo. It was observed that cashew embryos were found to be autonomy of growth regulator as the age increases and medium composition is only critical at very young age of the embryo. Successful germinated explants simultaneously produced shoot and root and were ready for transfer to field and acclimatization, between 90 and 112 days after inoculation.[...] Read More.

Keywords: Anacardium occidentale, in vitro culture, explant, embryo.

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Research Article

Somatic embryogenesis in two Iranian date palm cultivars

Parisa Eshraghi, Reza Zarghami*, and Mitra Mirabdulbaghi

Abstract: Shoot tips were removed from 3 to 4 year-old offshoots of adult date palm (Phoenix dactylifera L. cv. Khanizi and Mordarsing) and were cultured on medium that consisted of Murashige and Skoog basal salts medium. After 12 months they were transferred to three different treatments of growth regulators. Five months later, cv. Khanizi produced embryogenic callus on a medium containing 453 µM 2,4-dichlorophenoxyacetic acid (2,4-D), 15 µM N2- (2-isopentenyl) adenine (2ip), and 13 µM 6-benzyl amino purine. This callus was subcultured to another medium that was supplemented with 54 µM NAA and 148 µM 2ip. At this stage the embryos grew into plantlets. The cv. Mordarsing explants, on a medium containing 679 µM 2,4-D and 15 µM 2ip, produced embryogenic callus but the embryos remained in the globular stage.[...] Read More.

Keywords: Callus, in vitro tissue culture, offshoot, Phoenix dactylifera, regenerant.

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Research Article

Incorporation and Preservation of Newcastle Disease Virus on Filter Papers with Detection of Viral RNA via a Single-Tube RT-PCR Assay

P. N. Wambura*

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 19 (2), pp. 001-004, February, 2025. Available online at www.internationalscholarsjournals.org © International Scholars Journals

Full Length Research Paper

Incorporation and Preservation of Newcastle Disease Virus on Filter Papers with Detection of Viral RNA via a Single-Tube RT-PCR Assay

P. N. Wambura*

School of Veterinary Science, Faculty of Natural Resources, Agriculture and Veterinary Science, University of Queensland, Brisbane, QLD 4072, Australia. E-mail: p.wambura@mailbox.uq.edu.ac, phil_wambura@yahoo.com, pwambura@suanet.ac.tz.

Accepted 21 October, 2024

Suitability of storing infected allantoic fluid (AF) and cell culture supernatants (CCS) with strain I-2 of Newcastle disease virus (NDV) on Whatman filter papers at room temperature (22-25°C) and 37°C was determined. RNA was extracted from filter papers or liquid aliquots and subjected to reverse transcriptase- polymerase chain reactions (RT-PCR). The results showed that filter papers soaked with NDV infected AF or CCS stored at 37°C yielded amplicons with intensity similar to that kept at room temperature for up to 150 days. The study demonstrates that NDV infected samples can be soaked onto filter papers, stored and subsequently detected by RT-PCR. This method might be safely used for storage and transportation of NDV samples to the designated laboratories for molecular studies without the need for cooling.

Key words: Allantoic fluid, chicken embryo fibroblast, Newcastle diseases virus, polymerase chain reaction, RNA storage onto filter paper, strain I-2.

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Review

Manual Cloning of Mammals: An Innovative Methodology

Adil Salim l Elsheikh

Abstract: Handmade embryo reconstitution (HMER) has been used to study the parameters of nuclear transfer experiments such as fusion process, nuclear remodeling, nuclear reprogramming, biochemical processes, and biological processes during embryogenesis. These parameters have been widely investigated using the micromanipulator-based cloning technique (MBCT). This technique is a tedious, multi-step, time consuming and complicated procedure that utilizes expensive equipment. The HMER has emerged as an alternative for the MBCT. If the HMER is used to produce cloned animals it is known as handmade cloning (HMC). The HMC will allow the scientists to produce cloned animals with simple non-expensive equipment. Consequently, enormous data concerning all the facets of the nuclear transplantation experiments could be retrieved from various laboratories. This will allow a better future application of the cloning technique for the welfare of human, through production of animals with high genetic traits, rescue of endangered animal species and production of transgenic animals that can produce medicine for certain human diseases.[...] Read More.

Keywords: Cloning, chemical enucleation, oocyte bisection, mouse.

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Research Article

Competence and Survival of Lambs Following Transfer of Small Twin or Triple Embryos to Large Non-Prolific Recipient Sheep

S. M. K. Naqvi, A. Joshi, D. Kumar, R. Gulyani, V. P Maurya, S. Saha, J. P. Mittal and V. K. Singh

Abstract: The objective of the present study was to assess the embryo survival and development of progeny following transfer of either 2 or 3 embryos derived from dwarf size prolific Garole sheep into non-prolific large size Awassi x Malpura crossbred recipient ewes. Embryos were collected from donor ewes following induction of superovulation using FSH (5.4 mg Ovagen) and PMSG (200 IU) regimen. Estrus was synchronized in donor and recipient ewes by administering two injections of prostaglandin F2 a. The recipient ewes were divided into two groups and each recipient ewe received either 2 (Group 1) or 3 embryos (Group 2) of transferable quality in the uterine horn ipsilateral to corpus luteum. The recipient ewes of both the groups were examined for the presence of fetuses at 40 days of gestation by ultrasonography. The pregnancy and lambing percentages of ewes belonging to Group 2 were 57%, which was comparatively higher than Group 1 ewes where it was 42.9%. The survival of embryos was 38.1% in Group 2 and was higher compared to Group 1 (28.6%). The survival of lambs at weaning was higher in Group 1, compared to Group 2. The results indicate that survival of embryos and pregnancy rate was better following transfer of 3 than 2 embryos of prolific sheep to non-prolific sheep.[...] Read More.

Keywords: Microsheep, garole, embryo transfer, embryo survival, multiple births.

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Research Article

In Vitro Propagation of Miracle Berry (Synsepalum dulcificum) via Embryo and Nodal Cultures

K. E. Ogunsola* and C. O. Ilori

Abstract: Miracle berry is an evergreen tropical shrub which modifies sour food to produce a sweet taste. Its propagation is, however, hindered by seed recalcitrance and difficulty of stem to root. Thus in vitro propagation was investigated through embryo and nodal explants using different levels and combinations of auxins and cytokinins in MS medium. Embryo was regenerated in MS medium supplemented with 0.1 mg/l NAA + 0.2 mg/l BAP. Lateral buds proliferation was induced on the germinated embryo with 0.6 - 3.0 mg/l BAP + 0.1 - 0.2 mg/l NAA in which 3.0 mg/l BAP + 0.1 mg/l NAA produced highest number of buds. Rooting of the embryo regenerated plantlets was achieved with 1.0 - 2.0 mg/l IBA + 0.1 mg/l BAP. Very low (5 - 10%) axillary and terminal buds formation was achieved from nodal cultures. Few of the nodal explants formed buds with 0.1 - 0.8 mg/l NAA + 0.2 - 1.0 mg/l BAP + 0.02 mg/l GA3 with 0.8 mg/l NAA + 0.2 mg/l BAP producing the best result. However, all efforts to induce rooting on the buds formed from nodal explants proved abortive.[...] Read More.

Keywords: Miracle berry, in vitro conservation, recalcitrance, embryo culture, regeneration.

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Research Article

Regeneration of Kenyan Maize Inbred Lines and Their Single Cross Hybrids from Immature Embryos via Somatic Embryogenesis

Binott, J. J.,*, Songa, J. M., Ininda, J., Njagi, E. M. and Machuka, J.

Abstract: Field grown, self pollinated maize genotypes were planted in KARI (Kiboko and Kabete) research stations between January 2004 and May 2005. Immature maize embryos from twelve parental inbred lines and their respective single cross hybrids were evaluated for their ability form callus, somatic embryos and subsequent regeneration into plants. The embryos were excised from surface sterilized kernels harvested at different physiological stages, namely 10 - 24 days after pollination (DAP). They were used as explants to initiate callus on solid N6 basal media with varying level of 2,4-D (0 - 20 mg L-1) and regenerated on hormone free MS media. Optimal induction of primary callus at 2 mg L-1 averaged 83% and 67 in hybrids and inbred lines respectively. Somatic embryo competence was demonstrated in 6 inbreeds and 4 hybrids. However, plant regeneration was only achieved in 4 inbreeds and 3 hybrids. 90% percent of regenerants were normal and fertile. The successful regeneration of some of the inbred lines and/or hybrids provides a basis for development of genetic transformation using Agrobacterium tumefaciens to improve priority traits such as enhanced insects/pest and drought tolerance.[...] Read More.

Keywords: Inbred lines and hybrids, immature embryos, in vitro plant regeneration, recalcitrancy.

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Research Article

Embryo Production and Transfer in the Algerian "Cheurfa" Bovine Breed

Ferrouk Mustapha, Gharbi Ismail, Adel Djallal, Lafri Mohamed, Touati Kamel, Kaidi Rachid and Djamel Guetarni

Abstract: This work has permitted to test the response of the local cattle Cheurfa for a pFSH superovulation treatment based on administration of 40 mg pFSH (LH/FSH 40%), at a rhythm of 2 injections every 12 h between J10 and J13 of the oestrus cycle associated to injection of prostaglandin synthesis "Prosolvin®" at the 3rd day of the treatment. Two inseminations were carried out at 12 h interval after observed oestrus. The embryos were collected at J7. With four tests carried out, the average number of corpus luteum and collected embryos obtained were respectively 7.5 and 5 per cow. The number of transferable embryos was 2.33 per cow, with a viability rate of 46.66%. Five fresh embryos were transferred in recipients improved breed from the embryos obtained. The pregnancy rate obtained was 0% with 3 born calves Cheurfa type (2 male and 1 female). [...] Read More.

Keywords: Superovulation - embryo - transfer - cattle – local.

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Short Communication

Utilizing Transgenic Plants as Bioproduction Systems for Vaccines

B. Vinod Kumar, T. K. Raja, M. R. Wani, S. A. Sheikh, M. A. Lone, Gowher Nabi, M. M. Azooz, Muhammad Younis, Maryam Sarwat and Parvaiz Ahmad*

Abstract: Edible vaccine technology represents an alternative to fermentation based vaccine production system. Transgenic plants are used for the production of plant derived specific vaccines with native immunogenic properties stimulating both humoral and mucosal immune responses. Keeping in view the practical need of new technology for production and delivery of inexpensive vaccines, especially in developing world, plant derived edible vaccines is the best option in hand to combat infectious diseases. Plant derived vaccine is easy to administer, cost effective, readily acceptable, have increased safety, stability, versatility and efficacy. Several plant derived vaccines are under research, some are under clinical trials for commercial use. Like most biotechnology products, the IP situation for edible vaccines is complex as IP rights influence every stage of vaccine development. Keywords: Transgenic plants, edible vaccines, chimeric viruses, bacterial diseases, viral diseases. [...] Read More.

Keywords: Transgenic plants, edible vaccines, chimeric viruses, bacterial diseases, viral diseases.

2025, 24(1), 1-6;

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Research Article

Extraction of the NBS-LRR Class Resistance Gene (I2 Gene) from the Heamsona Tomato Cultivar

Pritesh Parmar* and R. B. Subramanian

Abstract: Three races of Fusarium oxysporum f. sp. lycopersici race 1, 2 and 3 are identified depending on the avirulence protein or effector protein secreted by fungal pathogen during the host colonization in tomato. These effector proteins are recognized by the host innate immune system based on R gene expressions that are I1, I2 and I3 in tomato for each races. Amongst the three, I2 protein has been cloned and characterized for the incompatibility against race 2 type of the pathogens. In India race 1 type of F. oxysporum f. sp. lycopersici observed commonly which require presence of I1 gene in tomato plant for the incompatibility reactions but in the present study, I2 gene was partially isolated from the tomato cultivar Heamsona and observed to be resistance against race 1 type of pathogen.[...] Read More.

Keywords: Fusarium wilt, race, R-gene, resistance, tomato.

2025, 24(1), 1-6;

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Research Article

Epidemiology of bacterial resistance and detection of metallo-β-lactamase (blaNDM) and imipenemase (blaIMP) resistance genes at CERBA

Géraud Joël GUIGMA*, Prosper BADO, Valérie Bapio BAZIE, Tampoubila Edwige YELEMKOURE, Serge Théophile SOUBEIGA, Delwende Leslie KABORE, Désiré ILBOUDO, Amana METUOR DABIRE, Albert Théophane YONLI and Jacques SIMPORE

Abstract: Received 6 January, 2025; Revised 28 February, 2025; Accepted 3 March, 2025; Published 7 April, 2025 Introduction: Antimicrobial resistance is a major public health concern. Producing new β-lactamases and carbapenemases is one form of resistance that preoccupies many scientists. The risk of the spread of carbapenemase-producing Enterobacteriaceae (EPCs) is a major public health issue, as these enzymes restrict therapeutic options and are often associated with other mechanisms, conferring multi-resistance on strains. Our study aimed to characterize the blaIMP and blaNDM resistance genes in Enterobacteriaceae isolates from urine cultures and genital swabs at CERBA from 2020 to 2023. Methodology: Pathogens were isolated on agar media, then identified using the API 20 E gallery; Imipenem-resistant strains were subjected to the traditional Hodge test to verify carbapenemase production. Detection of the IMP and NDM resistance genes coding for carbapenemases was carried out by multiplex real-time PCR at CERBA. Results: 1119 samples have been received for bacteriological analysis since January 2020. We noted 14.2% positivity to a clinically pathogenic strain. Bacterial species diversity was dominated by Escherichia coli in 54.71% of cases, followed by Klebsiella pneumoniae (15.72%). We observed a predominance of the NDM gene (97.9%) over IMP (2.1%). In some cases, we noted the coexistence of the IMP and NDM genes in Escherichia coli. Conclusion: This study enabled us to characterize the IMP and NDM resistance genes in isolation (IMP/NDM) or coexisting together (IMP+NDM) in Enterobacteriaceae isolates at CERBA. This study also enabled us to determine the frequency of bacterial species in bacterial culture samples at CERBA. [...] Read More.

Keywords: IMP, NDM, Enterobacteriaceae, β-lactamase, Carbapenemase.

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Research Article

Comparative Analysis of Escherichia Coli Concentrations in Shellfish Harvested from Different Sites on the Mediterranean Coast of Morocc

Rachid Boutaib, Hakima Azhari, Mohammed Abid and Mohammed Marhraoui

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 19 (4), pp. 001-008, April, 2025. www.internationalscholarsjournals.org/ © International Scholars Journals

Full Length Research Paper

Comparative Analysis of Escherichia Coli Concentrations in Shellfish Harvested from Different Sites on the Mediterranean Coast of Morocc

Rachid Boutaib1, Hakima Azhari1, Mohammed Abid2 and Mohammed Marhraoui1

1Institut National de Recherche Halieutique. Laboratoire de microbiologie. Centre Régional de l’INRH. BP. 5268. Dradeb.
Tanger 90000. Maroc.
2Institut Pasteur du Maroc. Département de recherche. 1, Plateau Marchan. Tanger 90000. Maroc

Accepted 11 November, 2016

Monitoring of ESCHERICHIA COLI levels in commercial bivalve shellfish is intended to protect consumer health from foodborne diseases. This study aims to identify if one species can be used as indicator for microbiological contamination of other species present. During 2012, 168 samples of shellfish (clams, cockles and mussels) were analysed for E. COLI by MPN technique. 62.5% of samples complied with regulatory threshold ≤230 MPN and geometric mean were higher in cockles than in clams or mussels in each site. Statistical significance was observed between sites. Seasonally, the highest levels were recorded in wet weather (winter and spring) due to runoff of waters from rainfall and lowest levels were recorded in dry weather (summer) when high temperatures show bactericidal effect. Our findings show cockles could be as sentinel specie for burrowing shellfish, but for non burrowing (mussels), monitoring should be done on this specie.

Keywords: Monitoring, Escherichia coli, Shellfish, MPN

2025, 24(1), 1-6;

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Review

Microbial Surfactants from Enterobacteriaceae: Production, Properties, and Environmental Applications

Alejandro De Jesús Cortés-Sánchez*, Mayra Diaz-Ramirez, Alan Javier Hernández-Álvarez, Felipe García-Ochoa, Adriana Villanueva-Carvajal, Liliana León-López, Alejandra L. San Martín-Azocar.

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 19 (4), pp. 001-010, April, 2025. www.internationalscholarsjournals.org/ © International Scholars Journals

Review

Microbial Surfactants from Enterobacteriaceae: Production, Properties, and Environmental Applications

Alejandro De Jesús Cortés-Sánchez1*, Mayra Diaz-Ramirez2, Alan Javier Hernández-Álvarez3, Felipe García-Ochoa4, Adriana Villanueva-Carvajal5, Liliana León-López6, Alejandra L. San Martín-Azocar7.

1Secretaria de Salud. Comisión Federal para la Protección contra Riesgos Sanitarios. Departamento de microbiología,
México.
2Departamento de Alimentos. División de Ciencias Biológicas y de la Salud. Universidad Autónoma Metropolitana (UAM),
Unidad Lerma. Edo. México.
3Food Research and Development Centre, Agriculture and Agri-Food Canada, 3600 Casavant Blvd. W. St. Hyacinthe,
QCJ2S 8E3, Canada.
4Departamento de Biofísica, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, México.
⦁ Facultad de Ciencias Agrícolas, Universidad Autónoma del Estado de México, Campus Universitario “El Cerrillo” A.P. 435,
Toluca, Estado de México C.P. 50200, México.
8Programa Regional del Posgrado en Biotecnología, Facultad de Ciencias Químico Biológicas, Universidad Autónoma de
Sinaloa, 80000 Culiacán, México.
⦁ Instituto Tecnológico y de Estudios Superiores de Monterrey, Escuela Superior de Ingeniería en Alimentos, Biotecnología,

Agronomía. Querétaro-México.

Accepted 08 October, 2016

In spite of being considered to be pathogenic microorganisms, some enterobacterial has the ability to produce biosurfactants (BS). These molecules besides their physicochemical and biological characteristics as emulsifiers, dispersants, stabilizers among others, are widely accepted in the industry since they are biodegradable, and present low toxicity. This has generated studies focused on the identification of new producer microorganisms, as well as the search of new carbon sources, improvement of production processes and the generation of a wide variety of patents. Their use is extensively spread in a diversity of industrial areas such as the pharmaceutic, food and environment, as alternatives of the surfactants derived from oil (which are highly pollutant),generating products and services of added value. In this review the properties and applications of the BS are summarized as well as the major genres of this group of Enterobacteriaceae that have been used for BS's production to update the actual advances on the benefits that these microorganisms possesses as BS producers.

Keywords: biosurfactants, enterobacterial, glycolipids, biotechnology, secondary metabolites.

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Research Article

Exploring Small Group Discussion as an Educational Strategy in Microbiology for Second-Year MBBS Students: Findings from a Pilot Study at Brims Bidar

Full Length Research Paper

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 19 (4), pp. 001-003, April, 2025. www.internationalscholarsjournals.org/ © International Scholars Journals

Full Length Research Paper

Exploring Small Group Discussion as an Educational Strategy in Microbiology for Second-Year MBBS Students: Findings from a Pilot Study at Brims Bidar

*Sudheendra Kulkarni1 and Chandrakanth Chillarge2

1Tutor and Nodal Officer for IDSP. Department of Microbiology, Bidar Institute of Medical Sciences, Bidar, Karnataka.
2Professor and HOD, Department of Microbiology Bidar Institute of Medical Sciences, Bidar, Karnataka.

Accepted 15 April, 2017

In India, Medical Microbiology subject is included in the curriculum of second year of MBBS. Teaching plays a major role in medical curriculum. There are various modes of teaching Medical microbiology such as lectures, tutorials, demonstrations, seminars, text book method, project method, small group discussions, study tours, problem solving method, team teaching, enquiry approach, videotapes, case studies etc. To teach a large group of students usually lectures are used. And to teach small group of students, demonstrations/ bedside clinics will be preferred. After thorough review of literature, we have found very few studies exist about the small group discussion as a teaching method in India. Hence we have undertaken the present study to observe the effectiveness of small group discussions as an effective teaching method in teaching Medical Microbiology.

Keywords: Microbiology, MBBS Students in Brims Bidar

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Research Article

Characterization and Bioactivity of Jatropha curcas Seed Lectin: Insights into Antifungal Applications

Mahmoud A. Al-Saman, Safinaz A. Farfour, Ahmed A. Tayel, Nashwa M. Rizk,

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 19 (4), pp. 001-011, April, 2025. www.internationalscholarsjournals.org/ © International Scholars Journals

Full Length Research Paper

Characterization and Bioactivity of Jatropha curcas Seed Lectin: Insights into Antifungal Applications

Mahmoud A. Al-Saman1,3, Safinaz A. Farfour2,3, Ahmed A. Tayel1,3, Nashwa M. Rizk2,3

1Industrial Biotechnology Department, 2Environmental Biotechnology Department, 3Genetic Engineering and Biotechnology Research Institute (GEBRI), University of Sadat City, Egypt.

Accepted 12 August, 2017

Lectins are non-immune, carbohydrate-binding proteins, which are very specific for sugar moieties and agglutinate cells or precipitate polysaccharides. Lectin from Egyptian Jatropha curcas seeds was isolated and purified using ammonium sulphate fractionation and gel filtration columns chromatography. The purified lectin has a specific activity of 351.5 hemagglutination unit/mg protein and a molecular weight of 28.00 kDa. The hemagglutination activity of lectin was suppressed by D-glucose, D-mannose and D-galactose. The amino acid composition of the purified lectin contained high ratio of acidic amino acids (38.42%), the hydroxy amino acids (11.10%), with minor amounts of sulfur containing amino acids and cysteine. The purified lectin from J. curcas seeds was heat - stable up to 60 ºC with trypsinized cattle erythrocytes and the hemagglutination activity decreased with increasing temperature. The purified lectin was insensitive from acidic to neutral conditions but was markedly affected by basic pH and the hemagglutination activity was encouraged by Mg2+, Ca2+ and Mn2+ ions. The purified lectin showed an antifungal activity against Fusarium oxysporum with a minimal inhibitory concentration of 70 µg / ml. However, lectin extraction from J. curcas could be recommended as a source for that could be applied as antifungal agent against phytopathogenic fungi.

Keywords: Agglutinin; Hemagglutination activity; Purification; Antifungal; Phytopathogens

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Research Article

Comparative Analysis of Silver Nanoparticles and Silver Nitrate: Antimicrobial Properties Against E. coli

Parameswari E., Udayasoorian C., S. Paul Sebastian and R.M. Jayabalakrishnan

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 19 (5), pp. 001-006, May, 2025. www.internationalscholarsjournals.org/ © International Scholars Journals

Full Length Research Paper

Comparative Analysis of Silver Nanoparticles and Silver Nitrate: Antimicrobial Properties Against E. coli

Parameswari E., Udayasoorian C., S. Paul Sebastian and R.M. Jayabalakrishnan

Tamil Nadu Agricultural University, Coimbatore, India

Accepted 17 September, 2024

Nanotechnology is expected to open new avenues to fight and prevent disease using atomic scale tailoring of materials. Among the most promising nanomaterials with antibacterial properties are metallic nanoparticles, which exhibit increased chemical activity due to their large surface to volume ratios and crystallographic surface structure. In this work we conducted batch experiments to assess the efficiency of silver nanoparticles synthesized by citrate reduction method for their antimicrobial property. The antimicrobial activity of silver nanoparticles and AgNO3 was compared in terms of ESCHERICHIA COLI growth rate, zone of inhibition and time dependent antimicrobial activity. Silver nanoparticles showed 100 per cent growth reduction of E. COLI when treated with 30 µg ml–1 concentrations, whereas the effect was much less at this concentration of AgNO3. Zone of inhibition test was also done for identification of degree of inhibition by using different concentration of AgNO3 and silver nanoparticles. It was found that, 10 µg ml-1 concentration was able to inhibit bacterial growth and created a zone of 0.8 cm by AgNO3 and 1.7 cm by Ag nanoparticles. Thus Ag nanoparticles are found to be efficient candidate for antimicrobial activity than AgNO3.

Key words: Antimicrobial potential, Silver nanoparticles, Escherichia coli.

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Research Article

Prevalence of Pathogenic Bacteria in Household Water: Implications for Water Management and Public Health in Lungwena

Taulo, S., *, Wetlesen, A., Abrahamsen, R., Mkakosya, R. and Kululanga, G.

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 19 (5), pp. 001-007, May, 2025. www.internationalscholarsjournals.org/ © International Scholars Journals

Full Length Research Paper

Prevalence of Pathogenic Bacteria in Household Water: Implications for Water Management and Public Health in Lungwena

Taulo, S.1, 3*, Wetlesen, A.1, Abrahamsen, R.1, Mkakosya, R.2 and Kululanga, G.3

1Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Science, P.O. Box 5003, N-
1432 As, Norway.
2Department of Microbiology, College of Medicine, P/B 360, Blantyre, Malawi.

3Faculty of Engineering, University of Malawi, P/B 303, Blantyre, Malawi.

Accepted 8 May, 2025

This study investigated and compared the microbiological quality of source, transported and stored water in Lungwena households. It also examined water management practices at all the investigated points. One hundred and eighty (180) water samples were collected from 6 villages and tested for Escherichia coli, Salmonella, E .coli 0157:H7 and Campylobacter jejuni using standard methods. Water contamination practices were observed in two hundred and eighty seven households. E. coli, Salmonella, E. coli 0157:H7 and C. jejuni were isolated in 54, 24, 6.7 and 2.2% of the samples, respectively. Sampling points revealed a significant difference (p = 0.001) in E. coli concentration. Salmonella concentration between sampling points was not significant (p > 0.05). E. coli concentration was significantly (p = 0.042) higher than that of Salmonella spp. The microbiological quality of water was found to be poor as a result of both poor water management practices and environmental sanitation. There were no significant differences (p > 0.05) in water management practices among the villages.

Key words: Pathogens, stored water, transport water, water contamination.

2025, 24(1), 1-6;

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Research Article

Intestinal Parasitic Infections Among Primary School Students in a Rural Region of Iran: A 2008 Study

Khosrow Hazrati Tappe, Habib Mohammadzadeh, Shahla Khashaveh, Baratali Rezapour and Afshin Barazesh*

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 19 (5), pp. 001-004, May, 2025. www.internationalscholarsjournals.org/ © International Scholars Journals

Full Length Research Paper

Intestinal Parasitic Infections Among Primary School Students in a Rural Region of Iran: A 2008 Study

Khosrow Hazrati Tappe1, Habib Mohammadzadeh1, Shahla Khashaveh1, Baratali Rezapour2 and Afshin Barazesh3*

1Department of Parasitology and Mycology, Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran.
2Department of Environmental Health, Faculty of Public Health, Urmia University of Medical Sciences, Urmia, Iran.

3Department of Microbiology and Parasitology, Faculty of Medicine, The Persian Gulf Tropical and Infectious Diseases Research Center, Bushehr University of Medical Sciences, P. O. Box 3631, Bushehr, Iran.

Accepted 1 March, 2025

Duplicate stool specimens from 405 primary school attending students of Barandooz-Chay region of Urmia district were tested for intestinal parasites. Besides, duplicate scotch tape slides were obtained and examined microscopically for Enterobius vermicularis and Taenia sp. eggs. A questionnaire containing demographic data was filled for every case, and the relationship between them and the parasitic infection was assayed. Overall, intestinal parasitic prevalence was 42.5%. Prevalence of Giardia lamblia, Entamoeba coli, Blastocystis hominis, Iodamoeba butschlii, Enterobius vermicularis and Hymenolepis nana was 20.5, 14.6, 13.3, 2.5, 10.6 and 0.2%, respectively. No statistic relation was proved between these infections, having tap water facility at home or family population (except for E. vermicularis), but there was a significant relationship between parents’ education level and family population. According to relatively high prevalence of parasitic infections in the study field, it is necessary to increase hygienic and educational measures.

Key words: Prevalence, intestinal parasites, primary school, Urmia.

2025, 24(1), 1-6;

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Research Article

Survival Strategies of Vibrio parahaemolyticus and Vibrio alginolyticus in Seawater: Impact of Long-Term Starvation on Virulence and Cellular Characteristics

Fethi Ben Abdallah,*, Ali Ellafi, Rihab Lagha, Héla Kallel and Amina Bakhrouf

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 19 (5), pp. 001-010, May, 2025. www.internationalscholarsjournals.org/ © International Scholars Journals

Full Length Research Paper

Survival Strategies of Vibrio parahaemolyticus and Vibrio alginolyticus in Seawater: Impact of Long-Term Starvation on Virulence and Cellular Characteristics

Fethi Ben Abdallah1,2*, Ali Ellafi1, Rihab Lagha1, Héla Kallel2 and Amina Bakhrouf1

1Laboratoire d’Analyse, Traitement et Valorisation des Polluants de l’Environnement et des Produits, Faculté de
Pharmacie Rue Avicenne, Monastir 5000, Tunisie.
2Unité de Fermentation et de Développement de Vaccins Virologiques, Institut Pasteur de Tunis, 13 Place Pasteur, 1002, Tunisie.

Accepted 1 March, 2025

In this study, we incubated Vibrio parahaemolyticus and Vibrio alginolyticus (marine food-borne pathogens bacteria) in seawater for 8 months to study their morphologic, proteomic and genetic responses to starvation. The atomic force micrographs of stressed strains showed a reduction of the cells size and an evolution to two coccoid-shape forms whose length is less than 0.4 m and between 0.5 and 1 m. Extracellular protein patterns and gelatinase profiles of stressed bacteria were also altered. Indeed, these modifications were manifested by the appearance and/or disappearance of bands as well as in the level of expression of certain proteins. In addition, we also searched for the presence of eight Vibrio cholerae virulence genes: toxR, toxS, toxRS, ctxA, zot, ace, toxT, and Virulence Pathogenicity Island (VPI) in the genome of investigated strains. The expression level of VPI gene studied by reverse transcriptase polymerase chain reaction was decreased, whereas the mRNA quantities of toxR, toxS, and ace in starved Vibrio remained stable.

Key words: Vibrio, seawater, alterations, morphology, proteins secreted, virulence gene expression, RT-PCR.

2025, 24(1), 1-6;

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Research Article

Antibiotic Resistance in Non-Typhoid Salmonella Isolates from Humans and Poultry in Egypt and Algeria

Abdelhakim Aouf, Yamina Messai, Mohammed S. Salama, Hala M. Aboushady, Mervat G. El-Anany, Souhila Alouache and Rabah Bakour*

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 19 (5), pp. 001-007, May, 2025. www.internationalscholarsjournals.org/ © International Scholars Journals

Full Length Research Paper

Antibiotic Resistance in Non-Typhoid Salmonella Isolates from Humans and Poultry in Egypt and Algeria

Abdelhakim Aouf1,2, Yamina Messai1, Mohammed S. Salama3, Hala M. Aboushady2, Mervat G. El-Anany4, Souhila Alouache1 and Rabah Bakour1*

1Laboratory of Cellular and Molecular Biology, Faculty of Biological Sciences, University of Science and Technology
Houari Boumediene, Algiers, Algeria.
2Microbiology Laboratory, Faculty of Science, Ain Shams University, Cairo, Egypt.
3Molecular Biology Laboratory, Faculty of Science, Ain Shams University, Cairo, Egypt.

4Microbiology Laboratory, Kasr Alainy Hospital, Cairo, Egypt.

Accepted 28 March, 2025

Seventy six non-typhoid Salmonella were isolated from both human and poultry in Egypt and Algeria and tested for their antibiotics resistance. The incidence of multiple antibiotics resistance was high. To study -lactams resistance mechanisms, double disk synergy test (DDST) with and without cloxacilline was used, results revealed the production of extended spectrum - lactamases (ESBLs) and cephalosporinase in seven and one human Egyptian isolates, respectively. The seven ESBL isolates were identified as Salmonella enterica serotype Poona and their molecular typing by ERIC-PCR revealed unrelated genetic patterns, indicating that these isolates are not clonal. The Cephalosporinase-ESBL- producing isolate was identified as S. enterica serotype Hadar. Polymerase chain reaction (PCR) with specific primers showed the presence of blaTEM and blaSHV genes, respectively, in all and four ESBL producers, and blaAmpC gene was detected in cephalosporinase-producing isolate. Genetic transfer by conjugation and plasmid profiles analysis showed that these genes and their resistance markers were transferable in association with plasmids of 60 kb for ESBLs and 64 and 3.2 kb for AmpC cephalosporinase.

Key words: Non-typhoid Salmonella, extended spectrum -lactamases, cephalosporinase, Egypt, Algeria.

2025, 24(1), 1-6;

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Research Article

Cold-Active Alkaline Protease from Stenotrophomonas maltophilia: Optimization and Detergent Industry Applications

Mohammed Kuddus* and Pramod W. Ramteke

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 19 (6), pp. 001-008, June, 2025. www.internationalscholarsjournals.org/ © International Scholars Journals

Full Length Research Paper

Cold-Active Alkaline Protease from Stenotrophomonas maltophilia: Optimization and Detergent Industry Applications

Mohammed Kuddus1* and Pramod W. Ramteke2

1Protein Research Laboratory, Department of Biotechnology, Integral University, Lucknow – 226026, India.
2Department of Biological Sciences, Sam Higginbottom Institute of Agriculture, Technology and Science, Allahabad –
211007, India.

Accepted 09 March, 2025

Biodetergents are preferred over the conventional synthetic detergents in view of their better cleaning properties, low energy input and the alleviation of pollution. The biodetergents derived from mesophilic/thermophilic organisms and also peroxide-based synthetic detergents require high temperature for their optimum activity. Thus, cold-active enzymes are very useful as they work at lower temperatures and do not require the input of energy. The purpose of the present study was the production optimization and purification of cold-active alkaline protease from a novel psychro-tolerant Stenotrophomonas maltophilia MTCC 7528 and its application as a detergents additive for cold washing. Psychro-tolerant proteolytic bacterium S. maltophilia MTCC 7528 was isolated from soil of Gangotri glacier, Western Himalaya, India that produced maximum protease (56.2 U/ml) at 20°C and pH 9.0 after 120 h incubation in shaking condition (120 rev/min). The purified enzyme has molecular weight of 75 kDa with maximum activity and stability at pH 10 and 20°C temperature. It showed excellent compatibility with commercial detergents with improved cleansing power at low temperature. The enzyme completely removed blood and grass stains and increases the reflectance by 26 and 23%, respectively. Enzyme-based detergents find a wide range of applications in laundry and textile industries. Cold- active alkaline protease from psychro-tolerant S. maltophilia may be a potential component to be used as a detergent additive for cold washing that will be beneficial to save energy as they work at lower temperatures.

Key words: Alkaline protease, biodetergent, cold-active enzymes, wash performance.

2025, 24(1), 1-6;

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Research Article

Targeting FAD2 Gene with RNAi for Enhanced Oleic Acid Composition in Transgenic Brassica napus

Tian Baoming, Sun Dandan, Lian Yuli, Shu Haiyan, Ling Hua, Zang Xin, Wang Bonan and Pei Zhenqiang

Advanced Journal of Microbiology Research ISSN 2736-1756 Vol. 19 (6), pp. 001-006, June, 2025. www.internationalscholarsjournals.org/ © International Scholars Journals

Full Length Research Paper

Targeting FAD2 Gene with RNAi for Enhanced Oleic Acid Composition in Transgenic Brassica napus

Tian Baoming1, Sun Dandan1, Lian Yuli2, Shu Haiyan1, Ling Hua1, Zang Xin1, Wang Bonan1 and Pei Zhenqiang1

1Bioengineering Department, Zhengzhou University, Zhengzhou, 450001, China.
2Chaoyang Teachers College, Chaoyang, 122500, China.

Accepted 1 April, 2025

Oleate- 12 desaturase (FAD2) is a key enzyme involved in the conversion of oleic acid (C18:1) into linoleic acid (C18:2). Brassica napus FAD2 gene was targeted for silencing by its RNAi gene under the control of a seed-specific napin promoter. This study aims to identify the transgenic plants, and analyze the level of BnFAD2 transcripts in addition to fatty acid profile in T3 seeds. As indicated by PCR and southern blotting analysis, a total of six transgenic plants were developed. Analysis performed by RT-PCR revealed the significant down-regulation of BnFAD2 transcripts in developing T3 seeds, which resulted in 13.90 to 32.20% increase of oleic acid composition in mature T 3 seeds. The data demonstrated that BnFAD2 gene was efficiently down-regulated and mediated by its RNAi gene, and oleic acid composition in transgenic rapeseeds was significantly enhanced.

Key words: RNAi, oleate- 12 desaturase (FAD2) gene, oleic acid, Brassica napus.

2025, 24(1), 1-6;

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