Suporn Katawatin

Separation of bovine spermatozoa proteins using 2D-PAGE revealed the relationship between tektin-4 expression patterns and spermatozoa motility

Poor semen quality has long been associated with bull infertility. However, the molecular basis in spermatozoa cells underlying the mechanisms of bull infertility remain unknown. The purpose of this study was to determine whether there is any protein in bovine spermatozoa related to semen quality. Semen samples from 18 Brahman bulls, 3 to 10 yrs of age, were assessed for semen quality in terms of spermatozoa motility and spermatozoa morphology. Spermatozoa extracts were separated using 2D-PAGE followed by staining with Coomassie blue. At least one duplicate gel was performed for each sample. Each gel was scanned with an ImageScanner System and analyzed for spots by ImageMaster 2D platinum software. The related protein spot(s) with semen quality was cut from the gel and identified by LC MS/MS. The results showed that at least 600 protein spots were detected in the spermatozoa extracts of the Brahman bulls. Of all these spots, there were 3 of 56 kDa at pI 6.4, 6.6 and 6.8 (Z(1), Z(2) and Z(3), respectively) that clearly showed different expression pattern among 18 Brahman bulls. Of 18 bulls (a) five showed the presence of spot Z(1) and Z(2) (pattern A) (b) one of spot Z(3) (pattern B) (c) five of spot Z(2) and Z(3) (pattern C) (d) one of spot Z(1) (pattern D) and (e) six of spot Z(2) (pattern E). Identification of spot Z(1), Z(2) and Z(3) by LC MS/MS had a similar result as matched to the tektin-4 protein of Bos taurus with a respective score of 171, 557 and 591. The statistical analysis of the 56 kDa protein patterns, tektin-4, indicated a significant effect on spermatozoa motility (P < 0.05) albeit non-significant on spermatozoa morphology. The bulls which showed pattern A had a higher percentage of spermatozoa motility than pattern E (P < 0.05) and not different from pattern C (P > 0.05). The statistical analysis also revealed that the presence of spot Z(1) had an effect on the percentage of spermatozoa motility (P < 0.01), whereas the presence of spot Z(2) and Z(3) had no effect (P > 0.05). The correlation coefficient between the relative protein content of spot Z(1) and the percentage of spermatozoa motility was 0.49. Our study demonstrates that the expression patterns of tektin-4 were a proxy for an effect on spermatozoa motility and consequently bull infertility. It may be that these protein patterns can be used as markers for improving bovine reproduction.

Milk protein profiles in response to Streptococcus agalactiae subclinical mastitis in dairy cows.

The objective of this study was to investigate the milk protein profiles of normal milk and those of milk during the course of subclinical mastitis, caused by natural Streptococcus agalactiae infection. Two-dimensional gel electrophoresis and liquid chromatography mass spectrometry were used to assess protein profiles and to identify the proteins. The results showed that S. agalactiae subclinical mastitis altered the protein profiles of milk. Following Mascot database matching, 11 and 12 protein types were identified in the milk collected from healthy and S. agalactiae subclinical mastitic udders, respectively. The distinct presence of the antibacterial protein cathelicidin-1 was detected in infected milk samples, which in turn was highly correlated to the severity of subclinical mastitis as represented by the milk somatic cell count (r = 0.616), but not the bacterial count. The protein profile of milk reveals changes in the host response to S. agalactiae intramammary infection; cathelicidin-1 could therefore serve as a biomarker for the detection of subclinical mastitis in dairy cows.

Association of BoLA-DRB3 alleles with tick-borne disease tolerance in dairy cattle in a tropical environment

Tick-borne disease is one of the most harmful tropical diseases in dairy production. Selection of dairy cows for tolerance to tick-borne disease is a challenging concept for dairy breeders in the tropics. The objectives of this study were (1) to detect specific tick-borne pathogen in cattle of different genetics and (2) to examine the polymorphisms of DRB3.2 alleles in Thai dairy cattle and find the allelic association with tick-borne disease tolerance. Specific primers to Anaplasma marginale (AM), Babesia bigemina (BG) and Babesia bovis (BB) were used to detect the infections by PCR. The results showed that the high proportion of infections were found in Bos indicus (Sahiwal, n=95) and crossbred Holstein × Zebu (75:25 Holstein:Zebu, n=101), compared to high Holstein fraction crossbreed (≥ 87.5% Holstein, n=187). The proportion of triple infections was also highly found in high Holstein fractions crossbreed. This study confirmed that Zebuine (Bos indicus) had a higher degree of tolerance, even when infected by tick-borne pathogens, compared to high Holstein fraction crossbred. The associated alleles of DRB3.2 for tick-borne pathogen infection tolerance were found: DRB3*14 and *41 were found to be tolerant to A. marginale; *14 to B. bovis; and *10 and *51 to B. bigemina. These tolerance alleles could be used as potential markers for selection in dairy genetic evaluation. The associated alleles for susceptibility were also found: *2 was found to be susceptible to A. marginale; *3 and *16 to B. bovis; and *20 to B. bigemina. These susceptibility alleles could be used as markers for culling, and selection favoring susceptibility alleles should be considered to maintain heterozygote advantage and pathogen-specific memories in the herd.

Genetic associations between milk fat‐to‐protein ratio, milk production and fertility in the first two lactations of Thai Holsteins dairy cattle

The aims of this study were to estimate, simultaneously, the genetic parameters of test-day milk fat-to-protein ratio (FPR), test-day milk yield (MY), and days-open (DO) in the first two lactations of Thai Holsteins. A total of 76 194 test-day production records collected from 8874 cows with 8674 DO records between 2001 and 2011 from different lactations were treated as separated traits. The estimates of heritability for test-day FPR in the first lactation showed an increasing trend, whereas the estimates in the second lactation showed a U-shape trend. Genetic correlations for FPR-DO and MY-DO showed a decreasing trend along days in milk (DIM) in both lactations, whereas genetic correlations for FPR-MY increased along DIM in the first lactation but decreased in the second lactation. Genetic correlations of FPR between consecutive DIM were moderate to high, which showed the effectiveness of simultaneous analyses. Selection of FPR in the early stage has no adverse effect on MY and DO for the first lactation but has a negative effect on MY and positive effect on DO for the second lactation. This study showed that genetic improvement of the energy balance using FPR, MY and DO with multi-trait test day model could be applied in a Thailand dairy cattle breeding program.

Effect of milk fat to protein ratio on genetic variance for milk yield in Thai tropical Holstein cattle

Abstract: The objective of this research was to investigate the optimum fat to protein ratio (FPR) in Thai tropical Holstein dairy cattle. First parity data consisting of 20 492 milk yields (MY) records for 24 891 cows for the period 2001 and 2011, were used in the analysis. The analysis used a random regression test-day animal model of third-order Legendre polynomials through the creation of a covariance function based on different FPRs. Variance components were estimated using the Bayesian method via the Gibbs sampling. The estimated heritability of MY in relation to FPR ranged from 0.19 to 0.27 with the pattern being similar to the genetic variances. Genetic correlations of MY at different FPRs were high at consecutive FPRs and then declined to negative in response to greater differences in FPR. Based on the results, it is concluded that the optimum FPR is in the range of 0.9 to 1.9, corresponding to the genetically controlled energy balance for MY in tropical Holsteins.