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Genetic Divergence of Thai Indigenous Pigs from Three Distinct Geographic Regions Revealed by Microsatellite Marker Analysis
(2023-02-01)
Chaweewan, Kamon
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Mahinchai, Prapas
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Kongsook, Sornchai
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Soponchit, Surasak
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Weerasamith, Phuree
Thai indigenous pigs (TIPs) are important genetic resources. Crosses with exotic pig breeds and wild boars may cause genetic losses. To date, the physical characteristics of TIPs have been inconsistent. The classification of TIPs by genetic information is needed to pursue an appropriate conservation program. In this study, the genetic diversity, cluster analysis, and phylogenetic relationship of TIPs were investigated using twenty-nine pig microsatellite markers. Blood samples were collected from TIPs from three regions of Thailand: north (NT, n = 118), northeast (NE, n = 61), and south (ST, n = 75). The mean total number of distinct alleles and the effective number of alleles per locus were 11.851 and 5.497, respectively. The mean observed heterozygosity (Ho) and mean expected heterozygosity (He) were 0.562 and 0.837, respectively. The F values of the microsatellite loci were positive under Hardy–Weinberg Equilibrium at p < 0.001, with overall mean values of Fis, Fit, and Fst of 0.247, 0.281, and 0.046, respectively. A total of 5, 5, and 17 private alleles were found at frequencies greater than 0.050 in the NT, NE, and ST pigs, respectively. Three optimal clusters (K = 3) were proposed within the TIP populations. Pigs from the NT and NE regions were mixed in two clusters, while members of the ST region were clearly separated. The phylogenetic tree confirmed that the pigs from NT and NE were each divided into two subgroups, while the pigs from ST were clustered into one group. A microsatellite analysis revealed the high genetic diversity of the TIP populations and confirmed the genetic divergence of the TIPs from the European and Chinese breeds. A genetic admixture of the TIP with the local wild boars was detected.
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Effect of postmortem storage on shear force value and calpain/calpastatin activity in longissimus dorsi muscle of rusa deer (cervus timorensis)
(2018-08-14)
Jantasaeng, Orapin
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Jirajaroenrat, Kanya
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Tuntivisoottikul, Kunya
The objective of this study was to determine the effects of aging period on shear force values and activities of calpain/calpastatin enzymes of the longissimus dorsi (LD) muscle from rusa deer (Cervus timorensis). The shear force values were measured with instron materials testing machine. Results showed that postmortem storage influenced meat tenderness with a highly significant reduction of shear force value from 9.01±0.83 kg/cm<sup>2</sup> at day 1 to 4.34±0.10 kg/cm<sup>2</sup> at day 21 (P<0.01). Determination of the enzyme activities indicated that the μ-calpain activity decreased significantly from 1.50±0.42 to 0.19±0.28 units/g of meat (P<0.01), whereas m-calpain activity (22.88±9.64 to 16.95±8.34 units/g of meat) and calpastatin activity (9.93±2.37 to 6.82±2.96 units/g of meat) slightly decreased (P>0.05). Shear force values were significant correlated with higher levels of μ-calpain (r=0.915) and m-calpain (r=0.758), respectively. However, the values were not significant related to the calpastatin activity (r=0.462). Therefore, this study confirmed that in the case of rusa deer, μ-calpain is a key factor controlling postmortem meat tenderness compared to m-calpain and calpastatin. The results suggested that storage of the rusa deer meat for 7 days is enough to achieve aging of the LD muscle, which may help to reduce the cost of meat production.
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Practical use of percoll density gradient centrifugation on sperm sex determination in commercial dairy farm in Thailand
(2016-06-01)
Promthep, Kornkanok
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Satitmanwiwat, Saranya
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Kitiyanant, Narisorn
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Tantiwattanakul, Pisit
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Jirajaroenrat, Kanya
Our experiment demonstrated an extension of Percoll® Density Gradient Centrifugation to use as a selection tool for sperm sexing developed for dairy farmer. We develop a simple protocol for bovine sperm sexing and used for artificial insemination (AI) and in vitro fertilization (IVF) for embryo transfer (ET). The quality of sperm after centrifugation in each layer were significantly different (P <0.01). There was no significant difference between percentage of live sperm after centrifugation and fresh sperm before centrifugation (P >0.05). The phenotype and swimming capacity of sperms in each layer were significantly different (P < 0.01). It was found that the most appropriate sperm in the layer after centrifugation and after quinacrine staining was between 65-70% Percoll®. X-bearing spermatozoa at this layer was 60.75% and motility was 95.86±0.46%. After insemination, the pregnancy ratio in the experiment was 40% which is similar to the percentage of normal AI. The number of female offspring in the experiment was 71.4% in selected sperm vs 50% in non-selected sperm AI. In the In Vitro fertilization experiment, sperm from our 7 layer Percoll® gradient concentration can fertilize and produce blastocyst embryo successfully. The simple 7 layer Percoll® gradient concentration protocol is appropriate for dairy farmer to do as a pre-AI step to increase their female offspring in their herd.
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Identification and characterization of a cellulase-encoding gene from the buffalo rumen metagenomic library
(2012-06-29)
Nguyen, Nhung Hong
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Maruset, Lalita
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Uengwetwanit, Tanaporn
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Mhuantong, Wuttichai
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Harnpicharnchai, Piyanun
Microorganisms residing in the rumens of cattle represent a rich source of lignocellulose-degrading enzymes, since their diet consists of plant-based materials that are high in cellulose and hemicellulose. In this study, a metagenomic library was constructed from buffalo rumen contents using pCC1FOS fosmid vector. Ninety-three clones from the pooled library of approximately 10,000 clones showed degrading activity against AZCL-HE-Cellulose, whereas four other clones showed activity against AZCL-Xylan. Contig analysis of pyrosequencing data derived from the selected strongly positive clones revealed 15 ORFs that were closely related to lignocellulose-degrading enzymes belonging to several glycosyl hydrolase families. Glycosyl hydrolase family 5 (GHF5) was the most abundant glycosyl hydrolase found, and a majority of the GHF5s in our metagenomes were closely related to several ruminal bacteria, especially ones from other buffalo rumen metagenomes. Characterization of BT-01, a selected clone with highest cellulase activity from the primary plate screening assay, revealed a cellulase encoding gene with optimal working conditions at pH 5.5 at 50 °C. Along with its stability over acidic pH, the capability efficiently to hydrolyze cellulose in feed for broiler chickens, as exhibited in an in vitro digestibility test, suggests that BT-01 has potential application as a feed supplement.
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Isolation of a gene encoding a cellulolytic enzyme from swamp buffalo rumen metagenomes and its cloning and expression in escherichia coli
(2012-01-01)
Cheema, Tanzeem Akbar
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Jirajaroenrat, Kanya
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Sirinarumitr, Theerapol
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Rakshit, Sudip K.
Ruminants are capable of hydrolyzing lignocellulosic residues to absorbable sugars by virtue of the microbial communities residing in their rumen. However, large sections of such microbial communities are not yet culturable using conventional laboratory techniques. Therefore in the present study, the metagenomic DNA of swamp buffalo (Bubalus bubalis) rumen contents was explored using culture-independent techniques. The consensus regions of glycosyl hydrolase 5 (GH5) family of cellulases were used as primers for PCR amplification. A full-length metagenomic cellulase gene, Umcel5B29, with a complete open reading frame (ORF) of 1611bp was identified. The similarity search analysis revealed that Umcel5B29 is closely related to the cellulases (73% to 98% similarity) of ruminal unculturable microorganisms, indicating its phylogenetic origin. Further analysis indicated that Umcel5B29 does not contain a carbohydrate binding module (CBM). Subsequently, Umcel5B29 was overexpressed in Escherichia coli. The recombinant enzyme worked optimally at pH 5.5 and 45C, a condition similar to the buffalo's rumen. However, the enzyme retained more than 70% of its maximal activity after incubation at <inf>p</inf>H 4-7 and more than 50% maximal activity after incubation at 30-60°C for 30min. These characteristics render Umcel5B29 as a potential candidate for the bio-stoning process of denim. © 2012 Taylor and Francis Group, LLC.

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