Open Access Original Research Article

Molecular Detection and Typing of Cervical Human Papillomavirus in a Selected Population in Bayelsa State: Comparison with the National and International Trend and Clinical Significance

Y. I. Oboma, A. A. Ngokere, Y. M. Tatfeng, S. I. Musa, I. A. Ibrahim, S. E. Itodo, A. B. Abdu

Biotechnology Journal International, Page 1-16
DOI: 10.9734/bji/2021/v25i230134

Background: The link between cervical lesions and human Papillomavirus (HPV) 16 and 18 is well established, but the magnitude of the risk of association and the importance of other high-risk hpv types is uncertain in Bayelsa state.

Aims: The study was aimed at detecting and typing of cervical hpv among selected subjects, establish the relationship between cervical dysplasia and hpv and also asses the level of knowledge of hpv, perception and attitude of women in Bayelsa State.

Materials and Methods: Questionnaires were used to assess the level of knowledge of hpv and cervical cancer. Papanicolaou stain for cervical cytology and Haematoxylin and Eosin stain used to study general tissue structure. Nested PCR was used to detect and multiplex PCR for typing. Results: The prevalence of hpv spectrum among participants was 52% while high risk hpv was 24%. Five (5) subtypes were identified. The subtypes identified were hpv 52(40.4%) most predominant, followed by 51(1.9%), 45(1.9%), 31(1.9%) and hpv 30 (9.6%). The age-specific prevalence showed a peak prevalence of 44.2% in the ages of 25-34 years and lowest in the age group (15-24years). Sequence alignment showed a single point mutation for hpv 45 and several points' mutation for hpv 52 at certain points of the sequenced nucleotides with Sequence no: 53CN12 and SeqH2011055303 and accession no: MG195999 and MG196000. HPV 52 was highest compared to previous studies, national and international. The level of knowledge on hpv infection and cancer of the cervix was low among subjects with a percentage score of 38.9%. Among other variables studied, life time sexual partners showed a statistically significant relationship in the prevalence of hpv (OR=0.02, P˂0.01). Perceived seriousness on hpv infection was high among hpv positive subjects compared with hpv negative participants ( =16.39, p ˂0.01).

Conclusion: The emergence of hpv 52 in the study area requires public health attention and thus urgent need for local hpv vaccines production.

Open Access Original Research Article

Construction of an Agrobacterium Mediated RNAI Genetic Transformation Vector Targeting the Replicase Gene of Indian cassava mosaic virus and Evaluation of Their Transformation Ability in Cassava Immature Leaf Lobes

T. Anuradha, P. Prabha

Biotechnology Journal International, Page 17-24
DOI: 10.9734/bji/2021/v25i230135

Cassava mosaic virus is one of the major problems affecting cassava industry in India. Currently there are no effective strategy to completely protect cassava from cassava mosaic viruses. In order to attain cassava mosaic virus resistance RNAi vectors targeting the replicase gene of Indian cassava mosaic virus is constructed in this study. Their efficiency to transform cassava immature leaf lobes were also studied here. Replicase gene of Indian cassava mosaic virus in Tamilnadu are cloned and sequenced. Conserved domains are identified and sub cloned to CSIRO RNAi vector system and transformation studies are done in immature cassava leaves. Two different RNAi vectors were constructed, utilizing a conserved 440bp of 5’ end of ICMV Rep (AC1) gene which also corresponds to a part of AC4 gene, and functions as a viral RNAi suppressor protein. The partial Rep gene of ICMV was cloned in sense and anti-sense orientations in the RNAi intermediate vector, pHANNIBAL. After cloning into pHANNIBAL, the cloned RNAi gene cassettes of ICMV is released and cloned into the binary vector, pART27, which contains the kanamycin-resistance gene as a plant selectable marker. In order to use hygromycin as a selection agent in cassava genetic transformation, RNAi–Rep gene cassettes of ICMV were cloned into pCAMBIA1305.2. These constructs were named pICR1 and pICR2 respectively. The Genetic transformation studies in cassava leaves done using pICR2 vector could generate PCR positive plants. An agrobacterium mediated replicase RNAi vector is developed and that can be transformed into cassava immature leaf lobes. Their efficiency to silence the Indian cassava mosaic virus should be studied further.

Open Access Original Research Article

Optimization of Selected Process Parameters Affecting Yield of Green Synthesized Silver Nanoparticles and Their Antibacterial Activity

F. A. Ekaji, C. O. Akujobi, S. I. Umeh

Biotechnology Journal International, Page 25-36
DOI: 10.9734/bji/2021/v25i230136

Aims: To optimize effects of selected process parameters affecting yield of green synthesized silver nanoparticles and their antibacterial activity.

Study Design: Study was designed with 3 factors Box Behnken Design (Minitab 17) and Response optimizer (Minitab 17) was used to determine optimum values of the factors.

Place and Duration of Study: Department of Microbiology, Federal University of Technology, Owerri, Nigeria, from March to November, 2020.

Methodology: After extraction by boiling, qualitative phytochemical analysis of leaves’ extracts of Ipeoma batatas, Commelina africana and Manihot esculenta was carried out. Following synthesis of silver nanoparticles as prescribed by Box Behnken design, yield of AgNPs was optimized with Response optimizer (Minitab 17). Then antibacterial activity of resulting AgNPs was tested against isolates of P. aeruginosa and E. coli.

Results: Extracts contained alkaloids, tannins, proteins and amino acids, flavonoids and phenolic compounds, but no sterols and cardiac glycosides. Optimum pH, temperature and time obtained with Response Optimizer resulted in 62.6%, 55.8% and 54.9% increase in yield of AgNPs, with leaf extracts of C. africana, M. esculenta and I. batatas respectively, compared to un-optimized conditions. Absorbance for resulting AgNPs peaked between 380 to 400 nm. Zones of inhibition (mm) of P. aeruginosa with AgNPs synthesized using extracts of C. africana, I. batatas and M. esculenta were 10, 10 and 9 respectively, under un-optimized condition, and 12, 10 and 8 respectively, for optimized conditions. Against E. coli, they were 11, 11 and 12 for AgNPs synthesized with extracts of C. africana, I. batatas and M. esculenta respectively, under un-optimized condition, and 13, 9 and 11 respectively, for optimized conditions.

Conclusion: Leaf extracts of C. africana, I. batatas and M. esculenta can be used in synthesizing AgNPs, with marked antibacterial activities. Box Behnken design is useful for optimization of effects of process parameters.

Open Access Original Research Article

Genetic Characterization of Domestic Apple Varieties from Tuzla Canton, BiH

Ensar Salkić, Besim Salkic, Amela Hercegovac, Aldijana Avdić, Azra Dorić, Emir Imširović

Biotechnology Journal International, Page 37-43
DOI: 10.9734/bji/2021/v25i230137

Aims: The basic precondition for apple breeding is the genetic diversity of varieties, which implies a large number of different, positive genes that enable adaptation to different weather conditions, resistance to new diseases and pests. One of the reliable sources of genetic diversity are indigenous varieties of Bosnia and Herzegovina. Their genetic identification is the first step in a process that has as its ultimate goal the collection of genetic material. The main goal of this research is to analyze the genetic variability of five indigenous varieties of apple in Tuzla Canton, Bosnia and Herzegovina in terms of contributing to the management of conservation and expansion of existing genetic resources, spreading this material through registered nurseries.

Study Design: The research included fiveautochthonous apple varieties: "Ovčji nos“ “Dobrić“, "Šarenika“, "Rančica“ and "Petrovača“.

Place and Duration of Study: Samples of young leaves were collected at the site of Donji Moranjci, City of Srebrenik,Tuzla Canton, Bosnia and Herzegovina in the spring of 2019.

Methodology: In order to determine the genetic diversity of five indigenous apple varieties were genotyped ten SSR(Simple Sequence Repeats) markers.

Results: No cases of synonyms or homonyms were found within the analyzed set.The results of the study indicate a pronounced differentiation, ie all five examined varieties represent unique genotypes.

Conclusion: The examined sets of genotypes possess significant genetic variability, which is important especially when we consider that a relatively small number of samples have been analyzed.

Open Access Original Research Article

Effect of Pre and Post Infection Administration of Zinc and Selenium on some Biochemical indices in Wistar Rats infected with Trypanosoma brucei

Timothy Tese, Kola Matthew Anigo, Alice Onyeche Jonah, Meshack Anthony Kugama, Tinok Andrew, Janet Larai Ibrahim, Raymond Arumona, William Denbon Genesis Chinten

Biotechnology Journal International, Page 44-52
DOI: 10.9734/bji/2021/v25i230138

Aims: The effect of pre and post-infection administration of zinc and selenium on Trypanosoma brucei brucei infection in wistar rats on some biochemical parameters were investigated.

Study Design:  The study was designed to evaluate the effect of pre and post infection administration of zinc and selenium on Trypanosoma brucei brucei infection in wistar rats.

Place and Duration of Study: The study was conducted at the Nigerian Institute for Trypanosomiasis Research (NITR), Kaduna and Department of Biochemistry, Ahmadu Bello University (ABU), Zaria, between January 2014 to March 2014.

Methodology: Fifty four (54) wistar rats were divided into nine groups of six (6) rats each. Groups III-IX were infected intraperitoneally with 1x103parasite load. Animals in group III were                       infected but untreated while the infected/treated groups received prophylactic (IV, V and VI) and therapeutic (VII, VIII and IX) treatment with zinc and selenium 50mg/kg and 10mg/kg body weight respectively.

Results: Results obtained showed that serum alkaline phosphatase (ALP), alanine amino transferase (ALT) and aspartate amino transferase (AST) activities in the infected-untreated control group were significantly different (P<0.05) and higher as compared to infected treated and normal control. Similarly, the two minerals significantly decreased (P<0.05) the levels of Urea and Creatinine in the infected treated when compared to the infected untreated rats, with the exception of creatinine value of the post-infected treated with selenium where there was no significant (P>0.05) decrease. On the other hand, a significant (P<0.05) increase in serum levels of total Protein, albumin and total bilirubin and no significant (P>0.05) difference in direct bilirubin was recorded in infected treated group compared to infected untreated and normal control.

Conclusion: It was concluded that administration of zinc and selenium ameliorate the infection-induced biochemical changes in tissues and organs associated with Trypanosoma brucei brucei infection in Wistar rats.