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Publication Open Access Influence of dietary protein and energy levels on sow backfat thickness(2012) Surasak Jittakhot; Dusit Laohasinnarong; Kampon Kaeoket; Mahidol University. Faculty of Veterinary Science. Department of Pre-clinical and Applied Animal Science; Mahidol University. Faculty of Veterinary Science. Department of Clinical Sciences and Public HealthThe objective of the present study was to determine the influences of dietary protein and energy levels on sow backfat thickness. The study was conducted by randomly measured backfat thickness in both pregnant and lactating stages of 10,401 sows from 7 commercial herds in Thailand. The pregnant sow diets contained 14.4% crude protein (CP) and 3,346 kcal metabolizable energy (ME)/kg. The lactating sow diet contained 18.8% CP and 3,660 kcal ME/kg. The percentage of dietary protein was positively correlated with backfat thickness (r = 0.036, P < 0.01) and the metabolizable energy of the diet was positively correlated (r = 0.053, P < 0.01). Upon, stepwise method of multiple linear regression analysis showed that the percentage of dietary crude fat (X, %) was a key factor affecting the thickness of sow backfat (Y, mm) with statistical significance. The regression equation was; Y = 14.4 + (0.2X), (R2 = 0.009, P < 0.01). Conclusion, a major nutritional factor that response for changing of sow backfat thickness was the dietary energy in sow diet.Publication Open Access Influence of dietary protein and energy levels on sow backfat thickness(2012) Surasak Jittakhot; Dusit Laohasinnarong; Kampon Kaeoket; Mahidol University. Faculty of Veterinary Science. Department of Pre-clinic and Applied Animal Science; Mahidol University. Faculty of Veterinay Science. Department of Clinical Sciences and Public HealthThe objective of the present study was to determine the influences of dietary protein and energy levels on sow backfat thickness. The study was conducted by randomly measured backfat thickness in both pregnant and lactating stages of 10,401 sows from 7 commercial herds in Thailand. The pregnant sow diets contained 14.4% crude protein (CP) and 3,346 kcal metabolizable energy (ME)/kg. The lactating sow diet contained 18.8% CP and 3,660 kcal ME/kg. The percentage of dietary protein was positively correlated with backfat thickness (r = 0.036, P < 0.01) and the metabolizable energy of the diet was positively correlated (r = 0.053, P < 0.01). Upon, stepwise method of multiple linear regression analysis showed that the percentage of dietary crude fat (X, %) was a key factor affecting the thickness of sow backfat (Y, mm) with statistical significance. The regression equation was; Y = 14.4 + (0.2X), (R2 = 0.009, P < 0.01). Conclusion, a major nutritional factor that response for changing of sow backfat thickness was the dietary energy in sow diet.Publication Open Access Trypanosoma evansi infection in mice and sow: cryopreservation of T. evansi, its infectivity and subsequent clinical signs and pathological findings(2011) Siriporn Tantawet; Bastiaan Willemse; Piyanant Taweethavornsawat; Kampon Kaeoket; Mahidol University. Faculty of Veterinary Science. Department of Clinical Science and Public HealthPublication Open Access Rapid single cell typing using SYBR® Green Real-Time PCR together with Melt Curve analysis for sex identification of porcine sperm(2014) Varaporn Korchunjit; Kampon Kaeoket; Yindee Kitiyanant; Tuempong Wongtawan; Mahidol University. Faculty of Veterinary Science. Laboratory of Cellular Biomedicine and Veterinary Medicine.; Mahidol University. Faculty of Veterinary Sciences. Semen Laboratory.; Mahidol University. Faculty of Veterinary Science. Department of Clinical science and Public Health.; Mahidol University. Faculty of Science. Department of Anatomy.; Mahidol University. Institute of Molecular Biosciences.; Mahidol University. Faculty of Veterinary Science. Department of Pre-clinic and Applied Animal Science.Identification of X or Y chromosome is a very useful technique to verify the sex of boar sperm, but common methods used in pig such as Fluorescence In situ Hybridisation (FISH) and whole semen Polymerase Chain Reaction (PCR) have some limitations. FISH is highly accurate, but time-consuming (>3 days). Whole semen PCR is faster than FISH (3-6 h), but not highly accurate (approximate methods). The objective of this study was to develop a fast and highly accurate protocol to identify sex of boar sperm. In the present study, our team developed an alternative sex identification protocol using single cell SYBR® green real-time PCR technique together with low resolution melt curve analysis. Primers specific for chromosome 1 and chromosome Y, a high performance KAPA SYBR® DNA polymerase and Rotor gene PCR platform were used. Male and female single white blood cells were used to calculate sensitivity and specificity. Single sperm was picked up under inverted microscope and transferred to 1 μl of lysis buffer, and real-time PCR was run according to the programmed protocol and analyzed with melt curve analysis. Results showed that our method was a fast (<50 min) accurate method with high sensitivity (95-99%) and specificity (100%) with low percentage of PCR failure (< 3%). Validation of this method using boar whole semen detected Y sperm at 52% and X sperm at 48%, which was comparable to the theory ratio of X and Y sperm (50:50) in semen. It may be concluded that the single cell SYBR® green real-time PCR technique together with melt curve analysis is fast and accurate that can be used to identify sex of boar sperm.Publication Open Access Effect of using supernatant for post-thawing solution and semen extender prior to insemination on sow reproductive performance(2010) Cholthida Kasetrtut; Kampon Kaeoket; Mahidol University. Faculty of Veterinary Science. Semen Laboratory.The objective of the present study was to determine the effects of using supernatant (seminal plasma: extender, 50:50 v/v) as post-thawing solution and semen extender on sow fertility. Fifteen sows were allocated into the following experimental groups: Groups A (control), sows (n=5) were inseminated with fresh semen, using a dose of 4x109 spermatozoa in 60 ml of Modena™ extender; Group B, sows (n=5) were inseminated with frozen semen (supplemented with 10 mM of L-cysteine), using a dose of 2x109 spermatozoa in 60 ml of supernatant (50% v/v of seminal plasma plus ModenaTM extender); Group C, sows (n=5) were inseminated with frozen semen (supplemented with 10 mM of L-cysteine), using a dose of 2x109 spermatozoa in 60 ml of Modena™ extender. All sows were inseminated twice using an intrauterine catheter depending on their weaning to oestrous interval (WOI). Pregnancy rate (PR), farrowing rate (FR), total number of piglet born (TNB) and number of piglet born alive (NBA) were recorded. In group A, the PR and FR were 100%, TNB and NBA were 7.8±3.9 and 7±3.9, respectively. For frozen semen, PR, FR, TNB and NBA in group B were higher than group C (100% versus 60%, 60% versus 0%, 6.0±2.7 versus 0, 6.0±2.7 versus 0, respectively). In conclusion, using supernatant (50% v/v of seminal plasma plus ModenaTM) as post-thawing solution and semen extender for artificial insemination in field condition improve sow fertility.Publication Open Access Effects of adding melatonin on the quality of frozen-thawed boar semen(2017) Natcha Thongrueang; Nutchanat Chaibangyang; Panida Chanapiwat; Kampon Kaeoket; Mahidol University. Faculty of Veterinary Science. Department of Clinical Science and Public HealthThe aim of this study was to study the effects of adding melatonin at different concentrations on the quality of cryopreserved boar semen. Semen samples (n = 6) were collected by hand-glove technique. Semen samples were diluted with Modena? and divided into 6 groups. According to the concentrations of melatonin at 0 (control, group A), 0.1 (group B), 0.5 (group C), 1.0 (group D), 1.5 (group E) and 2.0 mM (group F) to the lactose-egg yolk extenders used to freeze boar semen with traditional method. Progressive motility, viability and acrosome integrity were evaluated both before and after cryopreservation. The results showed that here was no significant difference in percentage of progressive motility among groups. However, a higher percentage of progressive motility was found in group D (39.17%). A higher percentage of sperm viability and acrosome integrity were found in group F (28.33%) and group F (34.50%), respectively. In conclusion, the present results suggest that adding melatonin between 0.1 and 1 mM during freezing yield a superior post-thawed semen qualities than freezing without melatonin.Publication Open Access Supplemental effect of varying L-cysteine concentrations on the quality of cryopreserved boar semen.(2010) Kampon Kaeoket; Panida Chanapiwat; Padet Tummaruk; Mongkol Techakumphu; Mahidol University. Faculty of Veterinary Science. Semen Laboratory.Cryopreservation is associated with the production of reactive oxygen species, which leads to lipid peroxidation of the sperm membrane and consequently a reduction in sperm motility and decreased fertility potential. The aim of this study was to determine the optimal concentration of L-cysteine needed for cryopreservation of boar semen. Twelve boars provided semen of proven motility and morphology for this study. The semen was divided into four portions in which the lactose-egg yolk (LEY) extender used to resuspend the centrifuged sperm pellet was supplemented with various concentrations of L-cysteine to reach 0 mmol L−1(group I, control), 5 mmol L−1(group II), 10 mmol L−1(group III) and 15 mmol L−1(group IV). Semen suspensions were loaded in straws (0.5 mL) and placed in a controlled-rate freezer. After cryopreservation, frozen semen samples were thawed and investigated for progressive motility, viability using SYBR-14/EthD-1 staining and acrosome integrity using FITC-PNA/EthD-1 staining. There was a significantly higher (P < 0.01) percentage of progressive motility, viability and acrosomal integrity in two L-cysteine-supplemented groups (group II and group III) compared with the control. There was a biphasic effect of L-cysteine, with the highest percentage of progressive motility, viability and acrosomal integrity in group III. In conclusion, 5 or 10 mmol L−1 was the optimum concentration of L-cysteine to be added to the LEY extender for improving the quality of frozen–thawed boar semen.Publication Open Access Distribution of spermatozoa in the reproductive tracts of sows after intra-uterine insemination using frozen-thawed boar semen(2012) Panida Chanapiwat; Kampon Kaeoket; Padet Tummaruk; Mahidol University. Faculty of Veterinary ScienceThe aims of the present study were to determine the number of spermatozoa in the reproductive tract of sows after intra-uterine insemination (IUI) with frozen-thawed (FT) boar semen and to investigate the influence of adding seminal plasma in the thawing medium on the sperm transport in the female reproductive tract. Fourteen multiparous sows were divided into 3 groups: group I (n= 5), insemination with 80 ml of extended fresh semen, group II (n= 4), insemination with 40 ml of FT semen diluted with ModenaTM extender and group III (n= 5), insemination with 40 ml of FT semen diluted with combination of ModenaTM and seminal plasma. All the sows were inseminated once with 2x109 motile spermatozoa at 36.1±2.8 hours after human Chorionic Gonadotropin (hCG) administration. The reproductive tract was collected from the sows at 12.4±2.2 hours after insemination and was divided into 5 parts, i.e. ampulla, cranial isthmus, caudal isthmus, utero-tubal junction (UTJ) and cranial uterine horn. All parts of the reproductive tract were flushed by phosphate buffer solution and the number of spermatozoa was determined using a Neubauer hemocytometer. Spermatozoa were found in all parts of the sows’ reproductive tracts at 12.4 hr after IUI using either fresh (group I) or FT boar semen (group II and III). Most of the spermatozoa were found in the UTJ (47.8%, 47.8% and 38.1% in group I, II and III, respectively, p> 0.05) and the caudal isthmus (27.2%, 26.4% and 28.1% in group I, II and III, respectively, p> 0.05). The total number of recovered spermatozoa in group III (409,420 sperm) tended to be higher than group I (286,750 sperm, p= 0.109) and II (287,000 sperm, p= 0.139). Number of spermatozoa in the cranial isthmus in group III (76,400 sperm) tended to be higher than that in group I (35,750 sperm) and II (33,333 sperm) (p> 0.05). It can be concluded that at 12.4 hr after IUI using FT semen, spermatozoa were found in all parts of the reproductive tracts of the sows similar to that using extended fresh semen. Supplementation of seminal plasma in the thawing medium of FT boar semen tended to increase the transportation of spermatozoa toward the cranial isthmus. This might be due to the suppression of PMN cells in the female reproductive tract and the reduction in cryoinjury of the FT boar sperm caused by seminal plasma.Publication Open Access Morphological changes and infiltration of immune cells in the endometrium of anoestrus gilt in relation to the ovarian appearance and serum progesterone.(2010) Yuttapol Teamsuwan; Kampon Kaeoket; Paisan Tienthai; Padet Tummaruk; Mahidol University. Faculty of Veterinary ScienceThe present study investigates morphological changes and distribution of the leukocyte subpopulation in the endometrium of anoestrus gilts in relation to reproductive cycles and serum progesterone (P4). Selected genital organs from 30 gilts culled due to anoestrus were examined. The genital organs were classified according to the ovarian appearance into 3 groups, i.e. inactive (n = 10); follicular (n = 10); and luteal phase (n = 10). Blood samples were collected prior to slaughter to determine serum P4. Seven tissue samples were randomly collected from the uteri of the gilts and were examined for histological structures, i.e. epithelial types and height, number of blood vessel, secretory vesicle and endometrial glands. Number of leukocyte subsets, i.e. lymphocytes, neutrophils, eosinophils, macrophages and plasma cells were counted. On average, age and body weight at culling of the gilts were 306.4±39.9 d (range 233-407 d) and 150.4±24.8 kg (range 104.0-205.5 kg). Lymphocyte was the most common immune cell in all tissue layers and in all stages of the reproductive cycle. Lymphocytes in glandular layer in the inactive phase was higher than in the follicular (p=0.02) and luteal phases (p=0.05). Neutrophils in both epithelial and subepithelial layers in follicular phases was higher than luteal and inactive phases (p<0.001). Eosinophil in subepithelium in the luteal phase was higher than inactive (p=0.004) and follicular phases (p<0.001). An increase in the serum P4 resulted in an increase number of uterine glands (p<0.001), a decrease number of lymphocytes in all tissue layers (p<0.05), a decrease number of neutrophils in subepithelial layers (p=0.03) and an increase in the number of eosinophils in subepithelial layers (p<0.001). In conclusion, the infiltration of the leukocyte subpopulation in the endometrium of anoestrus gilts is largely dependent on the ovarian function. Neutrophils and eosinophils were common immune cells in follicular and luteal phases, respectively.Publication Open Access Comparative study on six different long term commercial extenders for fresh boar semen(2010) Kampon Kaeoket; Titisa Srisowanna; Ubonrat Wichaidit; Panida Chanapiwat; Sukanya Manee-in; Mahidol University. Faculty of Veterinary Science. Semen LaboratoryIn Thailand, there are numerous commercial extenders for preservation of boar semen; however, no study has been carried out to compare these extenders in view of maintaining of sperm motility, plasma membrane integrity and acrosome integrity during cold storage. The aim of this study was to assess the percentage of sperm motility, HOST positive, viability and acrosome integrity of boar spermatozoa extended in Merck-III, Androstar®Plus, ModenaTM, NUTRIXcell®, VITASEM LD, Duragen and Dofu goldTM. Ejaculated from boars (n=6, one ejaculate from each boar) were collected and sub-samples (splited samples) were diluted (3x109 spermatozoa/100 ml) in the different extenders and stored for 10 days at 18ºC. On every second day (days 0, 2, 4, 6, 8, 10) after storage, the sperm parameters such as sperm motility, HOST positive, viability and acrosome integrity were evaluated. There were significant differences in characteristic of sperm motility and acrosome integrity between extenders. For instance, on day 4, Merck III (short term extender) and other long term extenders are able to maintain the semen qualities as claimed by the manufacturing. On day 8, the percentages of sperm motility, viability and acrosome integrity were highest (p< 0.01) in Androstar®Plus (72%) and ModenaTM (75%), Androstar®Plus (67%) and ModenaTM(65.6%), and Androstar®Plus (74.6%), Duragen (72.4%) and VITASEM LD (71.6%), respectively. In conclusion, changes in motility, viability and acrosome integrity during storage were affected by the extender utilized, however long term extenders maintained a high percentage (70%) of sperm motility (i.e. Androstar®Plus, ModenaTM and Duragen) and acrosome integrity (i.e. Androstar®Plus, VITASEM LD and Duragen) through 8 days of storage.