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Open Access Method Article

A Novel Test System for Genotyping rs43703016 Single-nucleotide Substitutions in the Bovine CSN3 Gene

Svetlana Kovalchuk, Arina Tagmazyan, Eugene Klimov

Annual Research & Review in Biology, Page 1-5
DOI: 10.9734/arrb/2019/v32i430090

Aims: Caseins are among the main milk proteins that determine many of its properties. Bovine kappa-casein (CSN3) is associated with the qualitative composition of milk, as well as with the quality of cheese obtained from this milk. The rs43703016 single-nucleotide substitution (g.88532332A>C; Asp148Ala) in exon 4 of the bovine CSN3 gene plays an important role in the production of quality hard cheeses. Various methods for the DNA testing of this substitution have been developed in the last three decades. Emergent DNA technologies provide an opportunity to modernize methods of genotyping single-nucleotide polymorphisms.

Results: We have developed and verified a method to differentiate A/C alleles of the rs43703016 substitution in the bovine CSN3 gene by real-time PCR using allele-specific fluorescent probes.

Conclusion: Our new method allows fast genotyping of animals, and may be used for selection of cows carrying the CC genotype, which determines good cheese-making properties of milk.

Open Access Original Research Article

Dried Pig Feces: Impacts on Growth, Haematology and Histology of Clarias gariepinus

Adebola Oluyinka Ajiboye, Funmilola Ajani, Benjamin O. Emikpe, Abiodun Bukola Adesanya

Annual Research & Review in Biology, Page 1-10
DOI: 10.9734/arrb/2019/v32i430089

Aims: Improper utilization and disposal of pig faeces in pig farms have been a great concern due to their hazards and risks they constitute to the environment. This situation calls for effective utilization of pig Feces for fish feed. This project was designed to provide information on safe level of Dried Pig Feces (DPF) on growth, histology and haematology of Clarias gariepinus.

Study Design: A twelve week feeding trial experiment was conducted using one hundred and fifty samples of C. gariepinus juveniles. The fish meal was replaced at different graded levels of DPF (0%, 25%, 50%, 75% and 100%) and was coded T1, T2, T3, T4 and T5 respectively.

Materials and Methods: Growth parameters, histology of vital organs and haematology of the fish samples were determined according to standard methods. Data obtained were analysed using ANOVA and Duncan multiple range test was used to separate the means.

Results: The Mean Weight Gain (MWG) revealed a significant (P<0.05) value in T3 (69.40 g), while 100% inclusion exhibited lowest MWG (21.07%). The Packed Cell Volume, Haemoglobin and Red Blood Cell values obtained decreased with increase in DPF inclusions, while an increase was observed in T4 (26.50%). White Blood Cell count was significant (P<0.05) in T3 (16,150±2.07 g/l) and decreased in T4(15,800±1.01 g/l) and T5(12,650±0.85 g/l) respectively. Histological results confirmed the absence of lesions in the heart, gills and the intestine of fish samples. Lesions in the liver were severed in T4 and T5.

Conclusions: The findings of this study revealed that 50% inclusion of DPF is the safe level for growth of C. gariepinus. Since the histological results revealed no lesions in all the vital organs, except the liver that had severe lesions, DPF can be recommended as an alternative feed ingredient.

Open Access Original Research Article

PCR Detection of Mixed and Zoonoses Malaria Using Plasmodium spp Dynein Light Chain (dlc-tctex) Gene

Maureen W. Kariuki, Elijah K. Githui, Andrew G. McArthur, Rashid A. Aman, Nyamu M. Njagi, Agatha C. Mwangemi, Lucy W. Kamau

Annual Research & Review in Biology, Page 1-12
DOI: 10.9734/arrb/2019/v32i430091

Novel gene targets are needed in accurate diagnosis of malaria. Previous studies show that the dynein light chains (dlc) in Plasmodium are uniquely conserved within the species, possibly due to their role as the cargo adptor moiety. This study aimed at the development of PCR assay for the detection of Plasmodium based on the (dlc-Tctex) as a genus and species-specific tool in malaria diagnosis. Multiple primers were designed based on Plasmodium spp dlc(Tctex) genes. The primers were applied on PCR to detect malaria on clinical samples and on laboratory maintained isolates of P. falciparum and P. vivax for human infecting species and P. knowlesi and P. cynomolgi for zoonoses infection involving primates. The amplified PCR fragments were gene cleaned and sequenced. BLASTn e-values output from the raw nucleotide queries supports that the genes are uniquely conserved.  Species-specific primers amplified P.  falciparum infections with no cross-reactivity to P. vivax, P. knowlesi or P. cynomolgi species. In this assay only 11 out of the 30 microscope positive malaria positive clinical blood samples were positive for PCR detection of P. falciparum infection. Primers designed for Plasmodium genus amplified the target band in all clinical malaria samples but also had another specific band amplification. This preliminary data demonstrate that a species-specific dlc(Tctex) PCR assay can be used for detection of P. falciparum and optimized genus primers can be applied to differentiate mixed malaria infections.

Open Access Original Research Article

Synthetic Seed Production as a Tool for the Conservation and Domestication of Celastrus paniculatus: A Rare Medicinal Plant

D. L. C. K. Fonseka, W. W. U. I. Wickramaarachchi, R. P. S. Madushani

Annual Research & Review in Biology, Page 1-8
DOI: 10.9734/arrb/2019/v32i430092

The black-oil tree (Celastrus paniculatus Willd) is a highly valued medicinal plant species belong to the Celastraceae family, known as Jyothishmathi in Ayurveda and Duhundu in Sri Lanka and grows as a perennial vine. It is an endangered medicinal plant species recorded in the red list of endangered fauna and flora of Sri Lanka in 1999. The seed oil of Celastrus paniculatus contains sesquiterpene alkaloids namely; celapagine, celapanigine, celapanine and celastrol, used in traditional system of medicine for various disorders and because of its high pharmaceutical value, plants are over exploited in natural habitats. Owing to poor seed germination and lack of successful vegetative propagation methods, domestication and commercial planting of this important medicinal plant species to meet the demand seems impossible. Therefore, it is of high importance to develop a reliable and efficient in vitro propagation to produce black oil plants for commercial use. In this study, it was attempted to produce synthetic seeds of Celestrus paniculatus via in vitro multiple shoot proliferation. Nodal segment explants were collected from freshly emerged age of sprouts, surface sterilized and cultured in Murashige and Skoog medium supplemented with different 6-benzylaminopurine (BAP) and Thidiazuron (TDZ) concentrations for shoot induction. The highest soot proliferation rate; 25 shoot tips/explant were observed with 0.1 mg/L TDZ. Induced shoot tips were used for synthetic seed production after encapsulating with BAP and a-naphthalene acetic (NAA) enriched sodium alginate. Shoot tip encapsulated beads produced with 4% sodium alginate were firm, clear, round and uniform in size and easy to handle. The influence of growth regulators (BAP and NAA) and storage period on the germination of encapsulated shoot tips was studied to evaluate the success of encapsulated shoot tips as a propagule. The beads germinated with 2 mg/L BAP and 0.2 mg/L NAA provided 80% in vitro germination percentage. Shoot tips of synthetic seeds remained green and healthy after storage at 5°C for a period of 8 weeks. Current findings suggest that encapsulated micro shoots (synthetic seeds) could be produced successfully, as the first step in domestication and conservation of Celastrus paniculatus. Further studies required on rooting of micro shoots, acclimatization and transferring of plantlets produced from synthetic seeds to in vivo conditions for domestication and conservation purposes.

Open Access Original Research Article

Novel Cathelicidin Antimicrobial Peptides from Paa robertingeri

Qinghua Luo, Huaiqing Deng, Mengguang Yin, Chen Chen, Jiang Zhou

Annual Research & Review in Biology, Page 1-10
DOI: 10.9734/arrb/2019/v32i430093

This study aimed to describe two cathelicidins (cathelicidin-PR1 and cathelicidin-PR2) from the skin of Paa robertingeri (Anura: Ranidae). The deduced mature peptides cathelicidin-PR1 and cathelicidin-PR2 were composed of 29 and 25 residues, respectively. Cathelicidin - PR1 has higher antimicrobial activity it could kill Gram-positive and Gram-negative bacteria and even some fungal species. Cathelicidin-PR1 exhibited more effective than AMP in antimicrobial activity against Pseydomonas maltophilia clinical strain. On the contrary, cathelicidin-PR2 had very weak antimicrobial activity. Furthermore, cathelicidin-PR1 and cathelicidin-PR2 exhibited very low hemolytic activity against human erythrocytes and little hemagglutinating activity. The results suggested that the cathelicidin-PR1 might serve as a template for developing novel antibiotics.

Open Access Original Research Article

Comparative Studies on the Effect of Fermentation on the Nutritional Compositions and Anti-nutritional Levels of Glycine max Fermented Products: Tempeh and Soy-Iru

T. R. Omodara, E. Y. Aderibigbe

Annual Research & Review in Biology, Page 1-9
DOI: 10.9734/arrb/2019/v32i430094

Aims: A comparative study of fungi and bacteria fermentation of soybean (Glycine max) was carried out to determine the effect of fermentation on the nutritional composition of their fermented products: tempeh and ‘soy-iru’.

Study Design: The experiment was carried out in the Department of Microbiology, Ekiti State University, Ado Ekiti, Nigeria, between August, 2017 and July 2018.

Methodology: Soybean was processed into ‘soy-iru’ (bacterial fermentation) and tempeh (fungal fermentation) and the microbial load, physico-chemical properties, proximate composition, levels of anti-nutritional components (trypsin inhibitor and phytic acid), anti-oxidants (total phenol, total flavonoid and DPPH), in-vitro protein digestibility and vitamins (A, B, C, D, and E) were analyzed.

Results: The microbial load, pH increased progressively during fermentation, while there was a decrease in the titratable acidity (TTA) of the two products. The protein(%), ash(%) and fat(%) contents of the Glycine max cotyledons increased from 29.56, 1.86 and 24.36 in unfermented substrate to 33.61, 2.21 and 26.90, respectively, after 24hrs of fermentation to produce tempeh. However, there was a reduction in the crude fibre(%) and carbohydrate(%) content from 2.94 and 41.29 in unfermented substrate to 2.53 and 32.57, respectively, after 24hrs of fermentation. Similar trends were observed during the production of ‘soy-iru’, however the change in proximate composition was not as significant as observed in tempeh. There was significant decrease in the trypsin inhibitor and phytic acid levels of the two products. The levels of anti-oxidants, vitamins B, D, E and protein digestibility increased significantly, in both bacterial and fungal-fermented products.

Conclusion: This research has therefore shown that fungal fermentation of Glycine max seeds into tempeh may be a better alternative to ‘soy-iru’ which was obtained from bacterial fermentation, because of the significant lower level anti-nutritional factors in the former.