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ABSTRACT: Antibody (immunoglobulin G (IgG), IgM or IgA) levels relative to ferritin in six foal sera (three male and three female) after birth (day 0 and 2, 6, 10, 20, 28, 36, 40, 52 and 56 weeks of age) were semi-quantitatively measured with normalization with antibody activity to ferritin in one adult horse serum. After addition of horse spleen ferritin to the serum sample, the complex formed between antibodies to ferritin in the serum and ferritin was co-immunoprecipitated using antibody to horse spleen ferritin. Antibody classes of the co-immnoprecipitate were detected with antibodies specific for horse IgG, IgM or IgA heavy chain. Six adult horse serum samples were found to have ferritin-binding activities in all immunoglobulin classes examined. Although ferritin antibody activities (IgG, IgM and IgA) were scant in the foal sera before sucking colostrum (day 0), their activities increased at 2 weeks of age. IgG antibodies showed a biphasic response and IgM antibody activity increased up to 40 weeks of age. Antibody (IgG, IgM and IgA) activities to ferritin in three colostrum samples were significantly higher than in adult horse serum samples. These results demonstrate that antibody to ferritin in foal serum is derived from colostrum after birth and is produced thereafter.
Animal Science Journal 04/2013; · 0.86 Impact Factor
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ABSTRACT: Objective-To test the usefulness of measuring pH and refractometry index, compared with measuring calcium carbonate concentration, of preparturient mammary gland secretions for predicting parturition in mares. Design-Evaluation study. Animals-27 pregnant Thoroughbred mares. Procedures-Preparturient mammary gland secretion samples were obtained once or twice daily 10 days prior to foaling until parturition. The samples were analyzed for calcium carbonate concentration with a water hardness kit (151 samples), pH with pH test paper (222 samples), and refractometry index with a Brix refractometer (214 samples). The sensitivity, specificity, and positive and negative predictive values for each test were calculated for evaluation of predicting parturition. Results-The PPV within 72 hours and the NPV within 24 hours for calcium carbonate concentration determination (standard value set to 400 μg/g) were 93.8% and 98.3%, respectively. The PPV within 72 hours and the NPV within 24 hours for the pH test (standard value set at 6.4) were 97.9% and 99.4%, respectively. The PPV within 72 hours and the NPV within 24 hours for the Brix test (standard value set to 20%) were 73.2% and 96.5%, respectively. Conclusions and Clinical Relevance-Results suggested that the pH test with the standard value set at a pH of 6.4 would be useful in the management of preparturient mares by predicting when mares are not ready to foal. This was accomplished with equal effectiveness of measuring calcium carbonate concentration with a water hardness kit.
Journal of the American Veterinary Medical Association 01/2013; 242(2):242-8. · 1.79 Impact Factor
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ABSTRACT: Despite enormous efforts, biochemical and molecular mechanisms associated with equine reproduction, particularly processes of pregnancy establishment, have not been well characterized. Previously, PCR-selected suppression subtraction hybridization analysis was executed to identify unique molecules functioning in the equine endometrium during periods of pregnancy establishment, and granzyme B (GZMB) cDNA was found in the pregnant endometrial cDNA library. Because GZMB is produced from natural killer (NK) cells, endometrial expression of GZMB and immune-related transcripts were characterized in this study. The level of GZMB mRNA is higher in the pregnant endometrium than in non-pregnant ones. This expression was also confirmed through Western blot and immunohistochemical analyses. IL-2 mRNA declined as pregnancy progressed, while IL-15, IFNG and TGFB1 transcripts increased on day 19 and/or 25. Analyses of IL-4 and IL-12 mRNAs demonstrated the increase in these transcripts as pregnancy progressed. Increase in CCR5 and CCR4 mRNAs indicated that both Th1 and Th2 cells coexisted in the day 25 pregnant endometrium. Taken together, the expression of immune-related transcripts suggests that immunological responses are present even before the trophectoderm actually attaches to the uterine epithelial cells.
Journal of Reproduction and Development 11/2012; · 1.46 Impact Factor
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ABSTRACT: Equine bone scintigraphy is usually performed with horses in standing position under sedation. However, swaying motion often leads to poor-quality images. To examine the usefulness of motion correction (MC) processing, equine bone scintigrams were evaluated using Scheffé's method of paired comparisons. A significant difference in evaluation scores was detected by analysis of variance (F test, P<0.01). According to all observers, Yardstick analysis scores were higher for images use of MC processing than for those no use of MC processing, for all parts. Overall scores of 5 observers were as follows: without MC 100% acquisition time (AT, lowest), use of MC with 25% AT, MC 50% AT, MC 75% AT, and MC 100% AT (highest). Thus, MC processing shortens AT in equine bone scintigraphy, and it contributes to a reduction in the external radiation exposure of nurses/technicians.
Journal of Veterinary Medical Science 10/2012; · 0.85 Impact Factor
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ABSTRACT: Zinc is one of the essential microelements involved in the regulation of enzyme activity, as well as metabolism of nucleic acid and proteins. There have been few reports on equine serum zinc concentrations during the training period, and little is known about the relationship between zinc levels and diseases in horses. In this study, we measured serum zinc levels in healthy Thoroughbred racehorses, as well as in other horses, under general disease or training conditions. The reference value for serum zinc levels in Thoroughbred horses was 41-79 μg/dl. There were no differences due to sex or age. Significant decreases in serum zinc levels were observed after training, but serum zinc levels did not vary with intensity of sweating. Serum zinc levels were lower in horses clinically diagnosed as having shipping fever (36.3 ± 2.7 μg/dl), fever (45.3 ± 3.0 μg/dl) and cellulitis (44.0 ± 3.4 μg/dl), as compared to control values (59.7 ± 9.7 μg/dl). They also tended to decrease in experimentally infected horses one day after inoculation. Changes in serum zinc levels reached nadir one day after surgical invasion, except for a horse that experienced complicating shock. These results suggest that zinc is a serological indicator of inflammatory status in Thoroughbred horses.
Journal of Veterinary Medical Science 08/2012; · 0.85 Impact Factor
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ABSTRACT: A total of 87 Thoroughbred horses and 10 ixodid ticks from a ranch in Hidaka district, Hokkaido were tested for tick-borne diseases. Using the indirect fluorescent antibody (IFA) method, 3.4, 92.0 and 97.7% of the horses showed antibody titers of ≥80 against Anaplasma phagocytophilum, Rickettsia helvetica, and Borrelia garinii, respectively. This is the first report of infection with the 3 pathogens in horses in Japan. Using PCR, DNAs from the peripheral blood of all horses were found negative with any Anaplasma, Rickettsia and Borrelia spp., while those from Haemaphysalis megaspinosa ticks were found positive for Anaplasma sp. closely related to A. phagocytophilum in Japan, and A. bovis. B. japonica was also detected in an H. flava tick for the first time.
Journal of Veterinary Medical Science 08/2012; · 0.85 Impact Factor
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ABSTRACT: The object of this study was to evaluate the usefulness of measuring the differences in the values of the serum total protein (DVSTP) concentration of foals and the refractometry index (DVRI) of the milk of dams before and after nursing of the colostrum for assessing failure of passive transfer (FPT) in foals. Serum samples from 31 foals were collected before the first nursing and at another 1 to 6 times between 4 and 24 hr after birth. The paired colostrum and milk samples were collected from 14 of their dams at the same time. Serum samples were analyzed for IgG concentration using a single radial immunodiffusion (SRID) test (98 samples) and total protein concentration using a temperature-compensating refractometer (98 samples). Colostrum and milk samples were analyzed for refractometry index (RI) using a Brix refractometer (71 samples). DVSTP concentration and DVRI were significantly correlated with serum IgG concentration. The negative predictive value (NPV) of DVSTP concentration for detecting serum IgG concentrations <400 and <800 mg/dl were 98.2 and 91.3%, respectively. Furthermore, the NPV of DVRI for detecting serum IgG concentrations <400 and <800 mg/dl were 97.3 and 96.3%, when the cutoff value is set to 6 and 10%, respectively. The results suggest that measurement of the DVRI is useful in assessing FPT as an initial "stall-side" screening test, because it is procedurally easier to perform, is more cost-effective than any commercially available test kit, and allows for rapid interpretation.
Journal of Veterinary Medical Science 06/2012; · 0.85 Impact Factor
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ABSTRACT: A Thoroughbred colt with ambiguous external genitalia was presented for clinical and histological examinations. The colt had a short penis, located between his hind limbs and faced backward. The measurements of luteinizing hormone, follicle stimulating hormone, testosterone and ir-inhibin showed a tendency to increase gradually from April. Both sex-determining region of the Y chromosome and amelogenin gene fragments were detected by PCR method. The cytogenetic analysis revealed the 63,XO/64,XY mosaic karyotype (ratio 83:17). In autopsy, immature symmetrical subcutaneous testes were found in the inguinal regions. The testes and the other accessory sex organs were histologically normal. These results add to our knowledge of chromosomal abnormality and information of disorders of sex development in horse.
Journal of Veterinary Medical Science 05/2012; · 0.85 Impact Factor
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ABSTRACT: The present study was conducted to elucidate the profile of circulating gonadotropins and gonadal hormones from birth to puberty and relationship between gonadal seasonality and hormonal secretion in both sexes of Thoroughbred horses. Spring-born colts (n=6) and fillies (n=9) were blood sampled weekly from jugular vein from birth to 60 weeks of age. Circulating FSH, LH, prolactin, testosterone, progesterone, estradiol-17β, and immunoreactive (ir)-inhibin were measured by radioimmunoassay. In both sexes, the steroid hormones levels were remarkably high at birth, rapidly dropped within a week and remained at the lower levels until the start of second spring after birth. Ir-inhibin was also high during the birth, remaining lowest during winter and again increasing towards the second summer. There was an increase in FSH concentration in foals during the first summer months after birth and in the next summer, the FSH concentration along with that of LH increased significantly. The seasonal increase in circulating prolactin was remarkable even in the first year, and no differences were noted between the two summers. These results clearly demonstrated that the hypothalamo-pituitary axis is already responsive to changes in photoperiod and secrete prolactin similar to adult horses, but pituitary gonadotrophs for FSH and LH secretion is less sensitive. When the values of these hormones in the second breeding season after birth were compared with adult values of the respective sex in the breeding season, no significant differences were observed, indicating that spring-born fillies and colts have already attained the stage of puberty at the second breeding season after birth.
Journal of Reproduction and Development 05/2012; · 1.46 Impact Factor
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ABSTRACT: Prevalence of equine gastric ulcer syndrome in 85 young Thoroughbreds was investigated. The presence of gastric ulcers was confirmed in 27.1% (23/85) of the horses by endoscopic examination. Sixty-two horses without gastric ulcers were allocated randomly to either the treated group (31 horses) or sham-dosed control group (31 horses) in order to investigate the efficacy of omeprazole oral paste in the prevention of gastric ulcers. At the second endoscopic examination conducted after 28 days of administration, only 1 horse in the treated group developed gastric ulcers, while 12 horses developed gastric ulcers in the control group. Based on these data, the efficacy of omeprazole in prevention of equine gastric ulcers in young Thoroughbreds during the training period was confirmed.
Journal of Veterinary Medical Science 04/2012; 74(8):1079-81. · 0.85 Impact Factor
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ABSTRACT: The purpose of this study was to demonstrate that a barren parous Thoroughbred mare with lactation induced by hormonal treatment can be introduced to an orphan foal at the same farm and that the mare can become pregnant after the end of the hormonal treatment. An additional purpose was to investigate the changes in the plasma concentrations of prolactin, estradiol-17β, progesterone, follicle-stimulating hormone, and luteinizing hormone before, during, and after hormonal treatment. The difference in body weight between the adopted foal and the control foals, which were at the same farm and raised by their natural mothers, was 17 kg at 24 weeks old, when the foals were weaned. However, the adopted foal and the control foals had almost the same weight at 35 weeks old and later. The first ovulation after hormonal treatment was confirmed 10 days after the end of hormonal treatment and then the normal estrous cycle resumed. Furthermore, the changes in plasma progesterone, estradiol-17β, follicle-stimulating hormone, and luteinizing hormone showed regular patterns after the first ovulation. Conception was confirmed in the fifth ovulation. Meanwhile, another study demonstrated that conception was confirmed in the first ovulation after hormonal treatment. The present study is the first to demonstrate the hormonal profiles during and after induction of lactation in a Thoroughbred mare. This approach is useful for solving the economic and epidemic problems of introducing a nurse mare to an orphan foal.
Journal of Veterinary Medical Science 04/2012; 74(8):995-1002. · 0.85 Impact Factor
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ABSTRACT: The purpose of this study was to observe the changes in body temperature before parturition using a wireless temperature monitoring device (WTMD) and to evaluate the usefulness of body temperature measurements using a digital rectal thermometer (DRT) and a microchip transponder thermometry device (MTTD) for predicting parturition in mares. The body temperatures using a WTMD at 0 hr and -1 hr were significantly different from those at the same time on Days 1-5 (P<0.01). The temperature differences between the morning of Day 0 and at -3 hr, -2 hr, -1 hr and 0 hr using the DRT and MTTD showed a significant drop compared with the temperature differences between the morning and evening of Days 1-5 (P<0.05). Furthermore, when the cutoff value of the temperature differences between the morning and other times was set to ≤0, the sensitivities of the DRT and MTTD in the evening of Day 0 and at -3 hr were 43% and 100% and 71% and 86%, respectively. The results suggested that monitoring the body temperature differences between morning and within 3 hr before the time of parturition is a valuable method for predicting parturition in mares. Conversely, this method would be more useful in predicting parturition when used in combination with other observations such as the mammary gland size and waxing of the teat ends because it has nearly a 20% probability of false-positive results prior to the day of parturition.
Journal of Veterinary Medical Science 02/2012; 74(7):845-50. · 0.85 Impact Factor
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ABSTRACT: Escherichia coli(E. coli) isolated from the uterus of a Thoroughbred mare with bacterial endometritis was used to evaluate the effect of progesterone (P(4)) on the immune response of mares. Peripheral blood mononuclear cells (PBMCs) were collected from 10 nonpregnant clinically healthy adult mares (range, 4-12 years) during diestrus, four Thoroughbreds and six Hokkaido native horses. Cell proliferation and expression of cytokine mRNA, including interferon (IFN)-γ, tumor necrosis factor (TNF)-α and interleukin (IL)-10, of PBMCs stimulated with E. coli and P(4) were examined in vitro. P(4) was shown to have significantly inhibited E. coli induced proliferation and expression of IFN-γ in PBMCs. These results indicate that P(4) inhibits the immune response to E. coli in mares.
Journal of Veterinary Medical Science 12/2011; 74(5):629-32. · 0.85 Impact Factor
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Teruaki Tozaki,
Fumio Sato,
Emmeline W Hill,
Takeshi Miyake,
Yoshiro Endo,
Hironaga Kakoi,
Hitoshi Gawahara,
Kei-ichi Hirota,
Yasuko Nakano, Yasuo Nambo,
Masahiko Kurosawa
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ABSTRACT: Myostatin is a member of the transforming growth factor-β family with a key role in inhibition of muscle growth by negative regulation of both myoblast proliferation and differentiation. Recently, a genomic region on ECA18, which includes the MSTN gene, was identified as a candidate region influencing racing performance in Thoroughbreds. In this study, four SNPs on ECA18, g.65809482T>C, g.65868604G>T, g.66493737C>T, and g.66539967A>G, were genotyped in 91 Thoroughbred horses-in-training to evaluate the association between genotype and body composition traits, including body weight, withers height, chest circumference, cannon circumference, and body weight/withers height. Of these, statistically differences in body weight and body weight/withers height were associated with specific genotypes in males. Specifically, body weight/withers height showed statistically significant differences depending on genotype at g.658604G>T, g.66493737C>T, and g.66539967A>G (P<0.01) in males during the training period. Animals with a genotype associated with suitability for short-distance racing, C/C at g.66493737C>T, had the highest value (3.17 ± 0.05 kg·cm(-1)) for body weight/withers height in March, while those with a genotype associated with suitability for long-distance racing, T/T, had the lowest (2.99 ± 0.03 kg·cm(-1)). In females, the trends in the association of body weight/withers height with genotypes were similar to those observed in males. As the SNPs are not believed to be linked to coding variants in MSTN, these results suggest that regulation of MSTN gene expression influences skeletal muscle mass and hence racing performance, particularly optimum race distance, in Thoroughbred horses.
Journal of Veterinary Medical Science 08/2011; 73(12):1617-24. · 0.85 Impact Factor
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ABSTRACT: A factor responsible for progression to pregnancy establishment in the mare has not been definitively characterized. To identify factors possibly involved in the establishment of equine pregnancy, the endometrium was collected from day 13 (day 0=day of ovulation) cyclic and day 13, 19 and 25 pregnant animals. From initial subtractive hybridization studies, a calcium regulating factor, Stanniocalcin-1 (STC1) mRNA, was found as a candidate molecule expressed uniquely in the pregnant endometrium. Endometrial expression of STC1 mRNA was noted on day 19 and was markedly increased in the day 25 gravid endometrium. STC1 protein was found in the extracts of day 25 gravid endometrium and immunochemically localized in the uterine glands. In addition, STC1 protein was detected in uterine flushing media collected from day 25 pregnant mares. High concentrations of estradiol-17 β (E(2)) were detected in day 25 conceptuses. E(2) levels were much higher in the gravid endometrium than in other regions, whereas progesterone levels did not differ among the samples from different endometrial regions. Expression of STC1 mRNA, however, was not significantly upregulated in cultured endometrial explants treated with various concentrations of E(2) (0.01-100 ng/ml) with or without 10 ng/ml progesterone. These results indicate that an increase in STC1 expression appears to coincide with capsule disappearance in the conceptus, and suggest that STC1 from the uterine glands likely plays a role in conceptus development during the pregnancy establishment period in the mare.
Journal of Reproduction and Development 12/2010; 57(2):203-11. · 1.46 Impact Factor
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ABSTRACT: The concentrations of branched-chain amino acids (BCAA; valine, leucine, isoleucine) were determined in plasma of 7 healthy thoroughbred mares and their foals from birth (0 week) to 24 weeks of age, using automated high-performance liquid chromatography. In foals, the concentrations of plasma valine were significantly high (p<0.05) at 16, 20 and 24 weeks. The concentrations of plasma leucine were significantly high (p<0.05) at 1 and 3 weeks. The concentrations of plasma isoleucine were significantly high (p<0.05) from 1 to 24 weeks. In mares, the concentrations of plasma valine were significantly high (p<0.05) at 16 and 24 weeks. The concentrations of plasma leucine and isoleucine were significantly high (p<0.05) at 16 weeks. It was clear that the concentrations of plasma BCAA in foals and mares were at different levels at various times after birth. Since mares and foals were kept in health during this study, we could get the base data of the concentrations of BCAA in plasma of healthy foals and mares from birth to 24 weeks.
Journal of Veterinary Medical Science 12/2009; 71(12):1623-8. · 0.85 Impact Factor
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ABSTRACT: To elucidate the relationship between steroidogenic hormones and developing adrenal glands, we investigated the immunolocalization of steroidogenic enzymes in equine fetal adrenal glands during mid-late gestation. Fetal adrenal glands were obtained from three horses at 217, 225 and 235 days of gestation. Steroidogenic enzymes were immunolocalized using polyclonal antisera raised against bovine adrenal cholesterol side-chain cleavage cytochrome P450 (P450scc), human placental 3beta-hydroxysteroid dehydrogenase (3betaHSD), porcine testicular 17alpha-hydroxylase cytochrome P450 (P450c17) and human placental aromatase cytochrome P450 (P450arom). Histologically, cortex and medulla cells were clearly observed in the three fetal adrenal gland tissue samples. P450scc and P450c17 were identified in cortex cells close to medulla cells and in some medulla cells in the fetal adrenal glands. P450arom was present in both cortex and medulla cells in the fetal adrenal glands. However, 3betaHSD was not found in any of the equine fetal adrenal gland tissue samples. These results suggest that equine fetal adrenal glands have the ability to synthesize androgen and estrogen, which may play an important physiological role in the development of equine fetal adrenal glands.
Journal of Reproduction and Development 11/2007; 53(5):1093-8. · 1.46 Impact Factor
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ABSTRACT: In this study, the expression patterns of inhibins, activins, insulin-like growth factor-I (IGF-I) and steroidogenic enzymes in equine placentae recovered during the latter two-thirds of gestation were examined. Concentrations of inhibin A and inhibin pro-alphaC in endometrial and fetal placental tissue homogenates were very low during the period examined, whereas these tissues contained high concentrations of activin A. In both maternal endometrial and fetal placental tissues, activin A levels decreased as pregnancy progressed. Expression of inhibin alpha-subunit was not observed in the placenta using either immunohistochemistry or in situ hybridization. Inhibin/activin betaA-subunit and its mRNA were confined to maternal endometrial glands, whereas immunopositive betaB-subunit was not detected in either endometrial glands or microcotyledons. Cytochrome P450 side chain cleavage enzyme was detected by immunohistochemistry in both endometrial glands and microcotyledons, whereas cytochrome P450 17alpha-hydroxylase/lyase was absent in these tissues. Immunopositive signals for 3beta-hydroxysteroid dehydrogenase and cytochrome P450 aromatase were localized in microcotyledons but not in endometrial glands. Immunohistochemistry revealed that IGF-I was highly expressed in microcotyledons around Day 130, and decreased as pregnancy progressed. Changes in the expression of IGF-I were correlated with the number of PCNA positive cells in the placenta. The present study demonstrated the presence and localized the site of expression of activin, IGF-I and steroidogenic enzymes in equine placental tissues during the latter two-thirds of gestation; the results suggest that activin and IGF-I may be involved in the regulation of placental development.
Domestic Animal Endocrinology 08/2006; 31(1):19-34. · 2.06 Impact Factor
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ABSTRACT: The relationship among plasma levels of immunoreactive (ir)-inhibin, inhibin A, inhibin pro-alphaC, FSH, estradiol-17beta and follicular growth were investigated during the normal estrous cycle in mares. Seven mares were used for two successive normal estrous cycles. Follicular diameters and ovulation were obtained by transrectal ultrasonography, and blood samples were collected daily by jugular venipuncture for hormonal assay. The results showed that inhibin A was inversely correlated (r = -0.59, p < 0.0001) with FSH indicating its contribution to negative feedback control of FSH secretions from the pituitary gland. Estradiol-17beta increased during the follicular phase reaching a peak (37.9 +/- 3.8 pg/mL) 2 d before ovulation. Estradiol-17beta was positively correlated (r = 0.78, p < 0.0001) with inhibin A. The high levels of inhibin A and estradiol-17beta were associated with the growth of the preovulatory dominant follicle and inversely correlated with FSH suggesting that both hormones are products of the large dominant follicles and were responsible for the decline in FSH secretion during the follicular phase of estrous cycle. In conclusion, an inverse relationship between inhibin A and FSH was clearly demonstrated indicating that inhibin A has a key role in the negative feedback control of FSH from the pituitary gland. In addition, inhibin A and estra-diol-17beta secretions were associated with the growth of the preovulatory dominant follicle and were positively correlated.
Endocrine 10/2004; 25(1):7-14. · 1.42 Impact Factor
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Yumiko Tanaka,
Hiroyuki Taniyama,
Nobuo Tsunoda,
Chandana B Herath,
Rie Nakai,
Hiromi Shinbo,
Natsuko Nagamine, Yasuo Nambo,
Shun-Ichi Nagata,
Gen Watanabe,
Nigel P Groome,
Kazuyoshi Taya
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ABSTRACT: To clarify the source of inhibins in equine female fetuses, concentrations of immunoreactive (ir-) inhibin, inhibin pro-alphaC, and inhibin A in both fetal and maternal circulation and in fetal ovaries were measured. In addition, the localization of inhibin alpha and inhibin/activin beta(A), and beta(B) subunits and the expression of inhibin alpha(A) and inhibin/activin beta(A) subunit mRNA in fetal ovaries were investigated using immunohistochemistry and in situ hybridization. Concentrations of circulating ir-inhibin, inhibin pro-alphaC, and inhibin A were remarkably more elevated in the fetal than in the maternal circulation between Days 100 and 250 of gestation. Fetal ovaries contained large amounts of ir-inhibin, inhibin pro-alphaC, and inhibin A. In contrast, these inhibin forms were undetectable in both the maternal ovaries and placenta. The inhibin alpha and inhibin/activin beta(A) and beta(B) subunit proteins were localized to enlarged interstitial cells of the equine fetal ovary. Expression of inhibin alpha and inhibin/activin beta(A) subunit mRNAs were also observed in the interstitial cells. We conclude that the main source of large amounts of inhibins in fetal circulation is interstitial cells of fetal ovary and is not of maternal origin. Furthermore, these inhibins may play some important physiological roles in the development of gonads in the equine fetus.
Biology of Reproduction 02/2003; 68(1):328-35. · 4.01 Impact Factor
