Camus Adoligbe

An Optimized Trichloroacetic Acid/Acetone Precipitation Method for Two-Dimensional Gel Electrophoresis Analysis of Qinchuan Cattle Longissimus Dorsi Muscle Containing High Proportion of Marbling

Longissimus dorsi muscle (LD) proteomics provides a novel opportunity to reveal the molecular mechanism behind intramuscular fat deposition. Unfortunately, the vast amounts of lipids and nucleic acids in this tissue hampered LD proteomics analysis. Trichloroacetic acid (TCA)/acetone precipitation is a widely used method to remove contaminants from protein samples. However, the high speed centrifugation employed in this method produces hard precipitates, which restrict contaminant elimination and protein re-dissolution. To address the problem, the centrifugation precipitates were first grinded with a glass tissue grinder and then washed with 90% acetone (TCA/acetone-G-W) in the present study. According to our result, the treatment for solid precipitate facilitated non-protein contaminant removal and protein re-dissolution, ultimately improving two-dimensional gel electrophoresis (2-DE) analysis. Additionally, we also evaluated the effect of sample drying on 2-DE profile as well as protein yield. It was found that 30 min air-drying did not result in significant protein loss, but reduced horizontal streaking and smearing on 2-DE gel compared to 10 min. In summary, we developed an optimized TCA/acetone precipitation method for protein extraction of LD, in which the modifications improved the effectiveness of TCA/acetone method.

Novel polymorphism of the bovine fat mass and obesity-associated (FTO) gene are related to backfat thickness and longissimus muscle area in five Chinese native cattle breeds

In this study, genetic variation of the fat mass and obesity-associated gene (FTO) was detected by PCRSSCP and DNA sequencing in 618 individuals from five Chinese indigenous cattle breeds, and their genetic effects on meat quality traits were evaluated. The results showed that a novel single nucleotide polymorphism C1071T was detected in exon 5 and the allelic frequencies for the C and T alleles of the five breeds were 0.666/0.334, 0.583/0.417, 0.631/0.369, 0.653/0.347 and 0.689/0.311, respectively. Animals with CT genotype had higher mean values for backfat thickness than those with CC or TT genotypes (P < 0.01). Individuals with CC or CT genotypes had higher longissimus muscle area than those with TT genotype (P < 0.05). The FTO gene may be a candidate gene for identifying differences in meat quality traits and therefore, could be applied to marker-assisted selection of native Chinese cattle breeds.

Back fat thickness and meat tenderness are associated with a 526 T→A mutation in the exon 1 promoter region of the MyF-5 gene in Chinese Bos taurus.

Qualitative trait loci (QTL) for growth and meat quality traits in cattle (Bos taurus) have been previously mapped to three chromosome regions, 0 to 30, 55 to 70, and 70 to 80 cM on chromosome 5. We evaluated the allele frequencies and gene-specific single nucleotide polymorphisms (SNPs) of bovine myogenic factor 5 (MyF-5) in the QTL regions and their associations with live weight and meat characteristics in indigenous Chinese cattle breeds. PCR-SSCP methodology showed a T>A mutation at 526 bp. Least square analysis revealed a significant association of this SNP with backfat thickness and meat tenderness (P < 0.05), while no significant association was found with live weight, loin eye height, loin eye area, rib area, or water holding capacity. Allele frequencies of MyF-5-A/B in the five breeds were 0.760/0.239, 0.752/0.247, 0.629/0.370, 0.715/0.284, and 0.750/0.250, for JiaXian red, Luxi, Nanyang, Qinchuan, and XiaNan crossbreed, respectively. The genotype distributions for these alleles in two of the Chinese cattle breeds (Luxi and Qinchuan) were not in Hardy-Weinberg equilibrium (P < 0.05); while those for the other three breeds (JiaXian red, Nanyang, and XiaNan) were in agreement with Hardy-Weinberg equilibrium (P > 0.05). The genotypic frequencies among all five cattle breeds showed moderate diversity (0.25 < polymorphism information content < 0.5). Based on our findings, we suggest that the MyF-5 gene influences back fat thickness and meat tenderness in Chinese Bos taurus. This SNP could be useful for marker-assisted selection for meat quality traits in these cattle.

Toxoplasma gondii infection in meat animals from Africa: Systematic review and meta-analysis of sero-epidemiological studies

Aim: Toxoplasma gondii is an ubiquitous apicomplexan parasite which causes toxoplasmosis in humans and animals. Felids especially cats are definitive hosts and almost all warm-blooded mammals, including livestock and human can serve as intermediate hosts. Food animals can be reservoirs for T. gondii and act as one of the sources for parasite transmission to humans. The objective of this study is to collect serological data on the prevalence of anti-T. gondii antibody, and risk factors for certain food animals from Africa to provide a quantitative estimate of T. gondii infection among these species from different African countries. Materials and Methods: Four databases were used to search seroepidemiological data on the prevalence of anti-T. gondii antibody in food animals between 1969 and 2016 from African countries. The search focused on data obtained by serologic test in food animals and meta-analyses were performed per species. Results: A total of 30,742 individual samples from 24 countries, described in 68 articles were studied. The overall estimated prevalence for toxoplasmosis in chicken, camel, cattle, sheep, goat, pig were 37.4% (29.2-46.0%), 36% (18-56%), 12% (8-17%), 26.1% (17.0-37.0%), 22.9% (12.3-36.0%), and 26.0% (20-32.0%), respectively. Moreover, major risk factor of infection was age, farming system, and farm location. Conclusions: A significant variation in the seroepidemiological data was observed within each species and country. The results can aid in an updated epidemiological analysis but also can be used as an important input in quantitative microbial risk assessment models. Further studies are required for a better and continual evaluation of the occurrence of this zoonotic infection.

Production of transgenic beef cattle rich in n-3 PUFAs by somatic cell nuclear transfer

ObjectivesOmega-3 polyunsaturated fatty acids (n-3 PUFAs) are beneficial to human health. However, the n-3 PUFAs contents of the livestock meat that we consume daily are relatively low. Utilization of transgenic technology to increase n-3 PUFAs contents in livestock may solve this problem.ResultsThe omega-3 fatty acid desaturase (FAD3), encoded by fat1 gene derived from Caenorhabditis elegans (C. elegans), converts omega-6 polyunsaturated fatty acids (n-6 PUFAs) to n-3 PUFAs. In the study, a plasmid containing the codon-optimized C. elegansfat1 gene (mfat1) was constructed and used to produce transgenic beef cattle by somatic cell nuclear transfer. Fourteen transgenic calves were obtained, and the ratio of n-6 to n-3 PUFAs in the transgenic calves decreased from 5.33: 1 to 0.95: 1 compared with negative controls.ConclusionsOur results demonstrated that the codon-optimized C. elegansmfat1 gene can be functionally expressed in the beef cattle and converts n-6 PUFAs to n-3 PUFAs.

Lack of an association between a single nucleotide polymorphism in the bovine myogenic determination 1 (MyoD1) gene and meat quality traits in indigenous Chinese cattle breeds.

The myogenic determination 1 (MyoD1) gene is a member of the MyoD gene family. It encodes for skeletal muscle-specific transcription factors containing highly conserved basic helix-loop-helix regions that perform important roles in the initiation, maintenance, and regulation of phenotypic traits. We investigated a new single nucleotide polymorphism (SNP) in the MyoD1 gene to evaluate whether this polymorphism affects meat quality traits in five Chinese indigenous cattle breeds, namely Qinchuan (QC), Xia-Nan (XN), Nan-yang (NY), Luxi (LX), and Jia-xian red (JXR). A C→G transversion at position 624 was detected in exon 1 of the MyoD1 gene; it causes an amino acid substitution ((624)serine/(624)cysteine). Least squares analysis showed that this SNP is not significantly associated with back fat thickness, eye muscle area, intramuscular fat, or marbling. The A/B allelic frequencies in the five breeds were 0.810/0.189, 0.779/0.220, 0.768/0.231, 0.820/0.180, and 0.801/0.198, respectively. Based on the χ(2) test, the genotype distributions of four cattle breeds (LX, NY, QC, and XN) did not agree with Hardy-Weinberg equilibrium (P < 0.05); one breed (JXR) did not deviate significantly from Hardy-Weinberg equilibrium (P > 0.05). The genotypic frequencies among all five cattle breeds showed moderate diversity (0.25 < polymorphism information content < 0.5). We concluded that the C624G SNP of the MyoD1 gene does not influence meat quality traits in indigenous Chinese cattle breeds; however, this SNP could be included in breed composition and population admixture analyses due to the marked differences in allelic frequencies among these five breeds.

A novel polymorphism of the GDF₁₀ gene and its association with body measurement traits in Chinese indigenous cattle.

Body measurement traits are known to play numerous important roles in the assessment of productivity and economic value. They are influenced by several factors, among which genetic factors are predominant. The gene GDF₁₀ is involved in skeletal morphogenesis and is associated with body measurement traits. It may be an important candidate gene for marker-assisted selection. We used the PCR-SSCP technology to examine a possible association of the single nucleotide polymorphism (SNP) (G142A) of the bovine GDF₁₀ gene with body measurement traits in 417 animals belonging to six different Chinese cattle populations: Xue long (Xl), Luxi (Lx), Qinchuan (Qc), Jiaxian red (Jx), Xianang (Xn), and Nanyang (Ny). In the Jx population, least squares analysis revealed significant effects on hip width, chest depth and chest circumference. The animals with the GG genotype had higher mean values than those with the GA genotype for all three traits. We conclude that the SNP of the GDF₁₀ gene could be a very useful genetic marker for body traits in Jx cattle reproduction and breeding.

Evidence of multiple mechanisms of alphacypermethrin and deltamethrin resistance in ticks Rhipicephalus microplus in Benin, West Africa

Ticks are obligate haematophagous arthropods, causing heavy losses in affected livestock. The objective of this study is to investigate phenotypic and genotypic resistance in Rhipicephalus microplus populations from Benin. Engorged female adult ticks were collected from cattle in two districts of Benin. Bioassays, biochemical and molecular tests were carried out on these ticks to determine the phenotypic, enzymatic and genetic status of resistance. Results of bioassays showed high resistance factors (RFu202f>u202f41). The molecular tests showing the presence of the domain II mutation and absence of the domain III mutation in the voltage-gated sodium channel gene. Biochemical tests showed increased activity of esterases, multifunction oxidases and glutathione transferases in resistant samples. Genotyping the samples showed high levels of heterozygous genotypes (73.36% and 63.30%) as compared to homozygous susceptible and resistant genotypes (23.3% and 10%) respective at Samiondji and Betecoucou. A correlation between phenotype resistance and presence of the domain II mutation at the voltage gated sodium channel gene was observed suggesting that this could be associated with resistance. Target site mutation and metabolic detoxification are mechanisms of resistance to pyrethroids in R. microplus tick populations from Benin.

C/EBPα gene as a genetic marker for beef quality improvement.

Intramuscular fat (IMF) or intramuscular triglycerides are interspersed throughout the skeletal muscles. The IMF, also called marbling, imparts meat with flavor and juiciness and is one of the core criteria for judging carcass value. The quantity of IMF is influenced entirely by genetics. Recently, understanding the underlying genetic bases of IMF has been a focus particularly in the beef industry. In this study, with the deep insights of ameliorating the beef quality by genetic means, the role of the CCAAT/enhancer binding protein alpha (C/EBPα) gene was investigated by over-expressing C/EBPα in bovine muscle stem cells (MSCs) to initiate the adipogenic program. Prior to this, bovine MSCs were isolated and induced to differentiate into adipocytes from cells that were exposed to dexamethasone isobutylmethylxanthine and indomethacin; the presence of insulin and fetal bovine serum was examined. Either ectopic expression of C/EBPα or treatment with dexamethasone and insulin induced the accumulation of fat droplets and the expression of adipogenic induction genes (LPL, PPARγ, C/EBPβ, and C/EBPδ). The expression levels of myoblast-related genes (MyoD, Myf5, and Pax7) were also measured to assess the accuracy of the differentiation process. This study provides evidence that the C/EBPα gene is essential for cattle adipose tissue growth and development. Hence, this finding can contribute to improving beef carcass quality.

Preliminary study on the tick population of Benin wildlife at the moment of its invasion by the Rhipicephalus microplus tick (Canestrini, 1888)

Background and Aim: Rhipicephalus microplus (Rm) is one of the most problematic livestock tick species in the world. Its rapid propagation and resistance to acaricides make it control difficult in the sub-region and Benin particularly. The aim of this work was to check its presence in wildlife and to confirm the possible role of reservoir wildlife may play in the propagation of the parasite. This will help to design more efficient control strategy. Materials and Methods: This study was conducted from February to March 2017 in the National Parks of Benin (Pendjari and W Park) and wildfowl’s assembly and selling point in Benin. Ticks were manually picked with forceps from each animal after slaughtering by hunters then stored in 70° ethanol. Collected ticks were counted and identified in the laboratory using the identification key as described by Walker. Results: Overall, seven species of ticks (Amblyomma variegatum, Boophilus decoloratus, Rm, Boophilus spp., Hyalomma spp., Rhipicephalus sanguineus, Rhipicephalus spp.) were identified on nine wild animal species sampled (Cane rat, wildcat, Hare, Doe, Cricetoma, Buffalo, Buffon Cobe, and Bushbuck and Warthog). The average number of ticks varies from 3 to 6 between animal species, 3 to 7 between localities visited, and 2 to 5 between tick species. However, these differences are statistically significant only for localities. Considering tick species and animal species, the parasite load of Rm and Rhipicephalus spp. is higher; the buffalo being more infested. The analysis of deviance reveals that the abundance of ticks observed depends only on the observed localities (p>0.05). However, the interactions between animal species and localities on the one hand and between animal and tick species on the other hand, although not significant, have influenced the abundance of ticks as they reduce the residual deviance after their inclusion in the model. Conclusions: This study reported the presence of Rm in wildlife of Benin and confirmed its role in the maintenance and spread of the parasites. It is, therefore, an important risk factor that we must not neglect in the epidemiological surveillance and ticks control strategies in the West African sub-region and particularly in Benin.