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- 저자 : Kifaro, G.C. (검색결과 3 건)
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Emmanuel George Kifaro,김미정,정승원,노진용,송창선,Gerald Misinzo,김상경 한국바이오칩학회 2022 BioChip Journal Vol.16 No.4
In recent decades “saliva” has emerged as an important non-invasive biofluid for diagnostic purposes in both human and animal health sectors. However, with the rapid evolution of molecular detection technologies, the limitation has been the lack of an efficient method for the facile amplification of target RNA from such a complex matrix. Herein, we demonstrate the novel application of hydrogel microparticles of primer-immobilized networks (PIN) for direct quantitative reverse transcription PCR (dirRT-qPCR) of viral RNA from saliva samples without prior RNA purification. Each of these highly porous PIN particles operates as an independent reactor. They filter in micro-volumes of the analyte solution. Viral RNA is captured and converted to complementary DNA (cDNA) through the RT step using covalently incorporated RT primers. The PIN with cDNA of the viral target will be ready for subsequent highly specific qPCR. Preceded by heat-treatment for viral lysis, we were able to conduct PIN dirRT-qPCR with 95% efficiency of the matrix (M) gene for influenza A virus (IAV) and 5’ untranslated region (5’ UTR) for chicken coronavirus spiked into saliva samples. The addition of reverse transcriptase enzyme (RTase) and 10% dilution of the matrix improved the assay sensitivity considerably. PIN particles’ compatibility with microfluidic PCR chip technology has significantly reduced total sample processing time to 50 min, instead of an average of 120 min that are normally used by other assays. We anticipate this technology will be useful for other viral RNA targets by changing the incorporated RT primer sequences and can be adapted for onsite diagnostics.
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Hozza, W.A.,Mtenga, L.A.,Kifaro, G.C.,Shija, D.S.N.,Mushi, D.E.,Safari, J.G.,Shirima, E.J.M. Asian Australasian Association of Animal Productio 2014 Animal Bioscience Vol.27 No.12
The aim of the experiment was to study the effect of feeding system on meat quality characteristics of Small East African (SEA) goats and their crosses with Norwegian ($SEA{\times}N$) goats finished under small scale farming conditions. Twenty four castrated goats at the age of 18 months with live body weight of $16.7{\pm}0.54kg$ from each breed (SEA and $SEA{\times}N$) were distributed in a completely randomized design in a $2{\times}3$ factorial arrangement (two breed, and three dietary treatments). The dietary treatments were; no access to concentrate (T0), 66% access to ad libitum concentrate allowance (T66) and 100% access to ad libitum concentrate allowance with 20% refusal (T100) and the experimental period was for 84 days. In addition, all goats were allowed to graze for 2 hours daily and later fed grass hay on ad libitum basis. Daily feed intakes were recorded for all 84-days of experiment after which the animals were slaughtered. Feed intake of T100 animals was 536 g/d, which was 183 g/d higher than that of T66 group. Supplemented goats had significantly (p<0.05) better feed conversion efficiency. The SEA had higher (p<0.05) hot carcass weight (8.2 vs 7.9 kg), true dressing percentage (54.5 vs 53.3) and commercial dressing percentage (43.3 vs 41.6) compared to $SEA{\times}N$. There was no significant difference (p>0.05) for dressing percentage and carcass conformation among supplemented goats except fatness score, total fat depots and carcass fat which increased (p<0.05) with increasing concentrate levels in the diet. Increasing level of concentrate on offer increased meat dry matter with subsequent increase of fat in the meat. Muscle pH of goats fed concentrate declined rapidly and reached below 6 at 6 h post-mortem but temperature remained at $28^{\circ}C$. Cooking loss and meat tenderness improved (p<0.05) and thawing loss increased (p<0.05) with ageing period. Similarly, meat tenderness improved (p<0.05) with concentrate supplementation. Shear force of muscles varied from 36 to 66, the high values been associated with Semimembranosus and Gluteobiceps muscles. The present study demonstrates that there are differences in meat quality characteristics of meat from SEA goats and their crosses with Norwegian breeds finished under small scale farming conditions in rural areas. Therefore, concentrate supplementation of goats of both breeds improves meat quality attributes.
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Genetic Characterization of Indigenous Goats of Sub-saharan Africa Using Microsatellite DNA Markers
Chenyambuga, S.W.,Hanotte, O.,Hirbo, J.,Watts, P.C.,Kemp, S.J.,Kifaro, G.C.,Gwakisa, P.S.,Petersen, P.H.,Rege, J.E.O. Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.4
Genetic diversity of sub-Saharan African goats was assessed using 19 microsatellite markers. Breeds were sampled from eastern Africa (Maasai, Kigezi, Mubende, North West Highland, Arsi-Bale), southern Africa (Ndebele, Pafuri) and West Africa (West African Dwarf, Maure, Djallonke). European breeds (Grisons Striped, Toggenburg), Asian breeds (Mongolian Cashmere, Bandipur) and a Middle East breed (Arab) were also included. The mean number of alleles per locus and average gene diversity ranged from 5.26$\pm$0.464 (Djallonke) to 7.05$\pm$0.516 (Mubende) and from 0.542$\pm$0.036 (Pafuri) to 0.672$\pm$0.031 (Ndebele), respectively. The between breeds variation evaluated using $$G_{ST}$$ and $\theta$ were found to account for 14.6% ($\theta$) and 15.7% ($$G_{ST}$$) of the total genetic variation. The $D_{A}$ measure of genetic distance between pairs of breeds indicated that the largest genetic distance was between Pafuri and Djallonke while the lowest genetic distance was between Arsi-Bale and North West Highland. A neighbour-joining tree of breed relationships revealed that the breeds were grouped according to their geographic origins. Principal component analysis supported the grouping of the breeds according to their geographic origins. It was concluded that the relationships of sub-Saharan African goat breeds were according to their geographical locations implying that the goats of eastern Africa, West Africa and southern Africa are genetically distinct. Within each sub-region, goat populations could be differentiated according to morphological characteristics.
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