Male pseudohermaphroditism in a complex malformed calf born with an acardius amorphus cotwin-a case report.

BACKGROUND: Male pseudohermaphroditism is a developmental anomaly wherein animals are genetically and gonadally male, but their internal and/or external genitalia resemble those of females. In cattle, pseudohermaphroditism is often accompanied by multiple severe malformations. To the best of our knowledge, this is the first report of male pseudohermaphroditism in a complex malformed calf born with an acardius amorphous cotwin. CASE PRESENTATION: This report describes the case of a three-day-old, male anurous Japanese Black calf born with an acardius amorphous cotwin, complete absence of the tail, agenesis of the anus, separate scrota, and umbilical hernia. Transthoracic echocardiography and computed tomography revealed serious malformations in the skeletal system and the circulatory, digestive, urinary, and genital organs. Necropsy revealed rectal atresia, immature testes, epididymis, and penis, but no male accessory gonads. Histological analyses revealed vaginal- and uterine-like tissues adjacent to or fused to the rectum. Fluorescence in situ hybridization detected X and Y chromosomes, and some cells presented two X-probe signals in the same nucleus. CONCLUSIONS: In contrast to the male genitalia, the female genitalia derived from the Müllerian ducts were difficult to detect by necropsy in the presented case. Many similar cases may be overlooked in clinical practice.

Cytobrush cytology patterns of early postpartum dairy cows.

Endometrial cytobrush cytology has been recommended as a reliable method for determining the percentage of polymorphonuclear leukocytes (PMN%) in cattle smears to diagnose cytological endometritis (CE). In this study, the clarity of cytobrush　cytological smears and the influence of different sample evaluation methods (number　and types of cells counted) on CE diagnosis were evaluated. Samples from 28 lactating　Holstein cows were collected weekly between 3 and 7 weeks postpartum. Smear clarity,　based on cell density, quality of cell morphology, and red blood cell contamination,　was significantly poorer at 3 weeks than between 5 and 7 weeks postpartum. Five　different cell counting methods (C100, C200, C300, C400, and C500) were used,　where 100-500 nucleated cells (endometrial epithelial cells, PMN consisting of neutrophils, eosinophils and basophils, lymphocytes, and macrophages) were counted. Agreement of diagnostic results for CE between C300 and C500 and between C400 and C500 was excellent at all observation times. In calculations of the PMN% based on whether the number of lymphocytes and macrophages were or were not excluded in the denominator, exclusion of these cells in the calculations did not affect the diagnosis of CE. While reduced clarity in earlier stage samples might interfere with the accuracy of cytobrush cytology, C300 can be recommended to determine the endometrial PMN%.

Comparison of diagnostic methods for uterine health in dairy cattle on different days postpartum.

BACKGROUND: Uterine health status is a key factor for dairy management. METHODS: The uteri of 28 Holstein cows were assessed weekly by three different methods: ultrasonography (UT), Metricheck device (MT) and cytobrush (CYT), during two weeks to seven weeks postpartum. RESULTS: The percentage of cows with an MT-positive, UT-positive and CYT-positive result decreased dramatically at three weeks, four weeks and five weeks postpartum, respectively. The agreement of the results was poor or weak within six weeks postpartum (κ=0.09-0.35), and only at seven weeks moderate or good agreement was achieved (κ=0.60-0.70). When comparing the results obtained by UT and MT, 30 per cent or more of the total cows had conflicting results from postpartum three weeks to six weeks. The uterine health of cows was re-evaluated by the combined usage of UT and MT referring to CYT. The highest sensitivity was achieved when positive individuals were identified as either MT-positive or UT-positive, whereas the highest specificity occurred when results were positive by both MT- and UT- methods. CONCLUSION: The accuracy of early postpartum uterine health status by three methods depended heavily on the time postpartum. The combined use of UT and MT is a practical candidate for cow-side monitoring method of uterine health.

A simple and practical short-term timed artificial insemination protocol using estradiol benzoate with prostaglandin F2α in lactating dairy cows.

To minimize costs and labor for short-term ovulation synchronization protocol, we developed one wherein each treatment-drug administration and timed artificial insemination (TAI)-was performed 24 h apart. The objective of the present study was to evaluate this short-term ovulation synchronization protocol in lactating dairy cows. Data were derived from 133 inseminations performed in 120 cows (32 primiparous and 88 multiparous), and the ovaries of these cows were scanned using ultrasound. The cows detected to have a functional corpus luteum (CL) received prostaglandin F2α (PGF) as a luteolytic agent. The cows were randomly assigned to two treatment groups: (1) treatment with estradiol benzoate (EB) 24 h after PGF treatment, and TAI 24-28 h after EB treatment (EB group); and (2) treatment with gonadotropin-releasing hormone agonist (GnRH) 56 h after PGF treatment, and TAI 16-20 h after GnRH treatment (GnRH group). As a luteolytic agent, either dinoprost (DP; 25 mg) or D-cloprostenol (DCLP; 0.15 mg) was administered intramuscularly in each treatment group. Pregnancy per AI (P/AI) was significantly higher in the DP- or DCLP-treated cows in the EB group when compared with their counterparts in the GnRH group (64.5% vs. 33.3%, P = 0.03 in the DP-treated cows and 51.1% vs. 27.3%, P = 0.04 in the DCLP-treated cows, respectively). Regarding parity, multiparous cows had greater P/AI in the EB group than in the GnRH group (52.8% vs. 26.7%, P = 0.01), whereas primiparous cows showed no significant intergroup difference (65.2% vs. 41.7%, P = 0.28). To conclude, the use of a convenient synchronization protocol comprising the administration of PGF and EB 24 h apart, rather than PGF and GnRH 56 h apart, has greater potential to improve pregnancy rates after TAI in lactating dairy cows given that a functional CL was accurately detected. This beneficial effect of the protocol using EB was clearly demonstrated in multiparous cows.

Plasma steroid hormone concentrations and their relationships in Suffolk ewes during gestation and parturition.

In this study, we measured plasma concentrations of progesterone, pregnenolone, estradiol, estrone, estrone sulfate, and cortisol and analyzed the correlations between these hormones during gestation in 13 Suffolk ewes, the main breed in Japan. Progesterone increased during gestation and decreased a few days before parturition; however, this pattern was different in samples with high progesterone concentrations (P4 spike samples). This P4 spike was associated with a high pregnenolone concentration. Apart from the P4 spike, the progesterone change was similar to that in other sheep breeds. Pregnenolone increased during gestation and decreased after parturition. A significant correlation between progesterone and pregnenolone was observed a few days before parturition. Estrone sulfate and estradiol concentrations increased during gestation, but estrone did not. They increased shortly before parturition, and then decreased immediately after parturition. At parturition, the correlation between estrone and estrone sulfate was significantly stronger. Moreover, a strong correlation between estrone sulfate and estradiol was observed after parturition. Cortisol did not change during gestation and increased shortly before parturition. The results showed steroid hormone dynamics in normal pregnant Suffolk ewes, which were mostly in line with those of other sheep breeds. It should be noted that high progesterone concentrations altered the typical patterns.

Relationship between the side of pregnancy and side of subsequent ovarian activity during the early postpartum period in lactating dairy cows.

After parturition, the ovary ipsilateral to the side of previous pregnancy exhibits lower functional activity than that exhibited by the contralateral ovary. The local inhibitory effects of the corpus luteum of the previous pregnancy, and/or the presence of a previous gravid uterine horn, may induce the ipsilateral suppression of folliculogenesis. We examined the influence of the side of previous pregnancy on ovulation and folliculogenesis, until completion of the third postpartum ovulation. The ovaries of 30 Holstein cows were scanned by ultrasonography, through the three postpartum ovulation sequences. No significant differences in the development of growing follicles, 5-8 mm in diameter, were detected between ipsilateral and contralateral ovaries. However, the total number of dominant follicles emerging ipsilaterally before the second postpartum ovulation were less than those emerging contralaterally (25 vs. 75%), and both the first and second ovulation occurred less frequently on the ipsilateral versus contralateral side (23 vs. 77% and 27 vs. 73%, respectively). Sequential observation in this study clearly indicated that the influence of the side of previous pregnancy persisted until the second postpartum ovulation, and this affected postpartum dominant follicle selection and ovulation, but not the development of growing follicles.

A case of a giant mass in the lumbar region of a newborn calf.

The present report describes a newborn calf with spina bifida that presented with a giant mass of the lumbar region, as well as subsequent gross, histological, and immunohistochemical examinations. A malformed Japanese black calf (estimated weight = 20 kg) was euthanized immediately after birth. A gross evaluation revealed a giant mass (approximately 60 cm × 30 cm × 15 cm) covered by the hair coat in the lumbar region and connected with the hair coat of the trunk. The mass surface was divided by a deep polygonal groove and externally resembled a lobulated kidney. Histology and immunohistochemistry revealed that the giant mass comprised a vessel, bronchiolus lined with cuboidal epithelium, and small alveolus. Bone bleaching revealed various abnormalities, including spina bifida, vertebral fusion, vertebral deformity, vertebral malformation, vertebral scoliosis, and coxal bone malformation. Following a suggestion that the giant lumbar region mass was occupied by lung tissue, this case was considered to involve an asymmetric conjoined duplicitas that resulted in a very rare dichotomous spondylosis malformation.

Incidence of double ovulation during the early postpartum period in lactating dairy cows.

In lactating cattle, the incidence of twin calving has many negative impacts on production and reproduction in dairy farming. In almost all cases, natural twinning in dairy cattle is the result of double ovulation. It has been suggested that the milk production level of cows influences the number of ovulatory follicles. The objective of the present study was to investigate the incidence of double ovulations during the early postpartum period in relation to the productive and reproductive performance of dairy cows. The ovaries of 43 Holstein cows (26 primiparous and 17 multiparous) were ultrasonographically scanned throughout the three postpartum ovulation sequences. The incidence of double ovulation in the unilateral ovaries was 66.7%, with a higher incidence in the right ovary than in the left, whereas that in bilateral ovaries was 33.3%. When double ovulations were counted dividing into each side ovary in which ovulations occurred, the total frequency of ovulations deviated from a 1:1 ratio (60.3% in the right side and 39.7% in the left side, P < 0.05). In multiparous cows, double ovulation occurred more frequently than in primiparous cows (58.8% vs. 11.5% per cow and 30.0% vs. 3.8% per ovulation, respectively P < 0.01). The double ovulators experienced more anovulatory repeated waves of follicles before their first ovulations than the single ovulators, which resulted in an extension of the period from parturition to third ovulation (81.5 days vs. 64.2 days, P < 0.05). In the multiparous cows, the double ovulators exhibited higher peak milk yield (P < 0.01) with lower milk lactose concentration (P < 0.05), indicating the prevalence of a more severe negative energy balance during the postpartum 3-month compared to the multiparous single ovulators. Our results showed that, regardless of their parity, double ovulation had no impact on the reproductive performance of the cows. Two multiparous cows that experienced double ovulation during the early postpartum period subsequently conceived twin fetuses. It can be speculated that the incidence of double ovulations during the early postpartum period partly contributes to the increased incidence of undesirable twin births in multiparous dairy cows.

River temperature drives salmon survivorship: is it determined prior to ocean entry?

Early life is believed to be a critical stage for determining survivorship in all fish. Many studies have suggested that environmental conditions in the ocean determine the fry-to-adult survival rate of Pacific salmon but few investigations have been conducted on the importance of the brief freshwater period during the seaward migration on overall survivorship. Here, we found that most of the variation in survivorship of hatchery-reared chum salmon (Oncorhynchus keta) was explained by river temperature during the fry stage, despite spending most of their life (approx. 99%) at sea. After the annual release of a constant number of fry, the number of fry moving through the river at a downstream location varied greatly. The number of returning adults was positively correlated with the number of fry moving downstream. This result suggests that most salmon mortality occurred prior to ocean entry, and that short-term mortality in the river is a key factor determining major fluctuations in total mortality. Although marine mortality is often invoked in the literature as a key factor determining total mortality of chum salmon, attention should also be paid to freshwater mortality to understand the population dynamics of this species.

The influence of feeding pattern on changes in plasma ghrelin in the Holstein cow.

We measured the plasma ghrelin and cortisol concentrations in non-lactating cows under fixed-time feeding conditions followed by an acute or gradual fasting treatment. During the 4 days before fasting, animals in Group 1 were fed a fixed amount of rations at 0800 and 1600 hr, and those in Group 2 were fed a gradually reduced amount. Thereafter, the plasma ghrelin concentrations of each animal were measured for 40 hr. The plasma ghrelin concentrations, which were low at the onset of fasting, increased before and after 0800 during fasting in Group 1, but not in Group 2. There were no significant differences in the plasma cortisol concentration within or between the groups. It was demonstrated that acute fasting induces elevation of the plasma ghrelin concentration, but that gradual fasting does not. This result suggests that fixed-time and fixed-quantity feeding caused a daily ghrelin rhythm in the cow and that this rhythm influenced changes in plasma ghrelin.

Release and systemic accumulation of heavy metals from preformed crowns used in restoration of primary teeth.

Preformed crowns for restoration of primary teeth are used in various treatments and are essential for restoring the crowns of primary molars. However, there are concerns that mechanical, chemical, and thermal stimulation may cause release of components of such crowns. We examined systemic accumulation of heavy metals associated with preformed crowns (3M Stainless Steel Primary Molar Crowns) used in primary tooth restoration. The participants were 37 children who had visited the Pediatric Dental Clinic of Tsurumi University Dental Hospital. They were divided into two groups: 22 participants without a history of being fitted with a preformed crown for primary tooth restoration (controls), and 15 participants with preformed crowns for primary tooth restoration. Analysis of hair samples showed a significant difference in the level of the trace element Cr - an important component of the preformed crowns - between children with and without preformed crowns, but no significant differences in Fe or Ni levels. Levels of the trace elements Ni, Cr, and Fe were within allowable ranges, indicating that these minerals were not likely to be harmful.

Comparison of periodontal health status and oral health behavior between Japanese and Chinese dental students.

A survey was carried out to compare periodontal health status and oral health behavior between Japanese and Chinese dental students. Subjects consisted of 118 students at Nihon University School of Dentistry and 92 students at the school of Stomatology, Nanjing Medical University. Saliva occult blood test was performed to classify whether subjects may have periodontal disease. Further questionnaires were given to evaluate different lifestyles and oral hygiene habit. The positive rate of the saliva occult blood test in Japanese dental students was 13.6%, and that of Chinese dental students was 43.5%. Bleeding from gingiva as a subjective symptom was as follows: Japansese 7.6%, Chinese 37.0%. Japanese dental students brushed for 13.5 min each day. The rate for Chinese students was 4.6 min. Use of interdental devices was as follows: Japanese 33.1%, Chinese 7.6%. Differences of periodontal disease rates between Japanese and Chinese dental students are thought to be differences in oral hygiene, indicating the need for improvements in hygiene measures in Nanjing City. The establishment and strengthening of oral hygiene education, including the importance of tooth brushing for prevention of periodontal disease, has been proposed.

Postpartum Massive Hematoma within the Broad Ligament of the Uterus in a Broodmare Possibly Caused by Rupture of the Uterine Artery.

A broodmare showed mild signs of abdominal discomfort and anemia after normal delivery. Ultrasonographic examination revealed a massive hematoma within the broad ligament adjacent to the uterine horn. Internal bleeding into the peritoneal cavity (hemoabdomen) was not seen. Following treatment, the clinical signs improved. Hemorrhage caused by rupture of the arteries within the broad ligament of the uterus may be a cause of hematoma. Prepartum and postpartum rupture of the arteries supplying the reproductive organs in the mare, which is not uncommon, can be fatal if severe hemoabdomen occurs. In the present case, the hematoma was considered to be tightly encapsulated between two serosal membrane layers of the broad ligament, and the membranes had remained intact. Thus, the serosal membranes did not split open, and massive bleeding into the peritoneal cavity did not occur. For this reason, the present broodmare avoided potentially fatal hemorrhagic shock.

Relationships between the conception rate of estrus synchronization using estradiol benzoate and CIDR (progesterone) and other parameters in holstein lactating dairy cows.

The purpose of this study was to determine the relationships between conception rate and other parameters before estrus synchronization with a Controlled Internal Drug Release Device (CIDR) and estradiol benzoate (EB). In the estrus synchronization program, animals were injected with 2 mg EB and then received a CIDR. Seven days later, the CIDR was removed and the animals were given an injection of Prostaglandin F(2alpha). Twenty-four hours later, they received an injection of 1 mg EB, and they were artificially inseminated 24 h after that. This program was applied to 258 Holstein cows in Tohoku-machi (Aomori, Japan). Blood was collected at the beginning of the program, and the conception rate was determined about 40 days after insemination. The relationships among conception rate, blood biochemical values, age, body condition score and days in milk were statistically analyzed to determine better conditions for cow conception. The conception rate of the cows in the high progesterone group (more than 1 ng/ml, P(4)+) was significantly higher than that of the low progesterone group (less than 1 ng/ml, P(4)-; 47.9% vs. 28.6% P<0.01). In the P(4)- groups, the serum phospholipid level was significantly higher in the conception group than in the non-conception group, and the same tendency was seen in the P(4)+ groups. Blood urea nitrogen (BUN), albumin (Alb), and total cholesterol (TChol) were significantly higher in the conception group compared with the non-conception group, but no with P(4) was observed. We concluded that 1) the conception rate of the P(4)- group was remarkably low, that 2) the low conception rate and low P(4) level was related to a low PL level and that 3) BUN, Alb and TChol were higher in the conception group, although no relation with P(4) was found.

A solution for universal classification of species based on genomic DNA.

Traditionally, organisms have been classified on the basis of their phenotype. Recently, genotype-based classification has become possible through the development of sequencing technology. However, it is still difficult to apply sequencing approaches to the analysis of a large number of species due to the cost and labor. In most biological fields, the analysis of complex systems comprising various species has become an important theme, demanding an effective method for handling a vast number of species. In this paper, we have demonstrated, using plants, fish, and insects, that genome profiling, a compact technology for genome analysis, can classify organisms universally. Surprisingly, in all three of the domains of organisms tested, the phylogenetic trees generated from the phenotype topologically matched completely those generated from the genotype. Furthermore, a single probe was sufficient for the genome profiling, thereby demonstrating that this methodology is universal and compact.

[An evaluation of virtual waxing up system with haptic interface].

PURPOSE: The purpose of this experiment was to construct a system of waxing pattern using virtual reality in the odontological field. The success of operation with feedback on the maniphalanx sense of waxing is reported. METHODS: The constructed device was a system based on PHANToM DESKTOP (SensAble Technologies Inc.). This device includes six-degrees-of-freedom input and reproduction of the sense of touch by anti-power feedback. The software which controls this device enables four operations such as "Cutting down", "Piling up", "Melting", and "Finishing" to be applied to the virtual waxing pattern by virtual reality. Finally, the virtual waxing was examined by applying this system to a virtual abutment tooth. RESULTS: With six-degrees-of-freedom input and anti-power feedback, a virtual waxing and the designer's highly developed designs were successfully expressed regardless of the virtual space. CONCLUSIONS: 1. The virtual waxing up could be done by six-degrees-of-freedom input with the stylus in a virtual space. 2. The time taken for all these numerical values of corresponding rate of volume, occlusal, mesio-distal and bucco-lingual views to reach 100+/-5%ranged from 50 to 75 minutes, with an average of 63 minutes. 3. The virtual wax improvement showed the tendency to trace the passage of first arranging the volume, and straightening the outlook on the occlusal view at the end. 4. This system, which provides an interface between man machines, six-degrees-of-freedom input and anti-power feedback, has much in common with the existing waxing up and crafting. The demand for special technologies by technical persons and engineers can be kept to a minimum and application of the method can be expanded into education and to objective evaluations.

Relationship between seminal plasma lactoferrin and gonadal function in horses.

Total 78 semen samples were obtained from 27 Thoroughbred stallions (aged 6 to 27 years), and were subjected to quantification of lactoferrin (Lf) in seminal plasma and examination of the seminal properties. The seminal plasma Lf concentration varied from 21 to 689 microg/ml, with a mean value of 244 +/- 151 microg/ml (S.D.). The seminal plasma Lf concentration and total seminal plasma Lf positively correlated with the sperm concentration (r=0.5938, P<0.001) and with the total sperm number (r=0.6959, P<0.001), respectively. There was no correlation between seminal plasma Lf and sperm motility. These results suggest that seminal plasma Lf reflects gonadal function.

Seminal plasma lactoferrin but not transferrin reflects gonadal function in dogs.

Lactoferrin purified from canine seminal plasma by a three-step chromatography procedure had a molecular mass of 75.2 kDa and cross-reacted with antiserum to equine seminal plasma lactoferrin. Seminal plasma lactoferrin concentrations were determined by a competitive enzyme-linked immunosorbent assay (ELISA) by using rabbit anti-equine lactoferrin antibody and alkaline phosphatase-labeled goat anti-rabbit IgG antibody in 14 normal dogs and found to range from 12 to 197 micro g/ml, with a mean value of 77 +/- 59 micro g/ml (the mean +/- SD). Seminal plasma transferrin concentrations were determined by a sandwich ELISA with goat antibody to canine serum transferrin and alkaline phosphatase-conjugated goat anti-canine transferrin antibody and found to range from 0.32 to 12.6 micro g/m l, with a mean value of 2.44 +/- 3.25 micro g/m l. The lactoferrin concentration significantly correlated with the sperm concentration (r=0.7025, P<0.01), but there was no significant correlation between the seminal plasma transferrin concentration and sperm density. These results indicate that seminal plasma lactoferrin, but not transferrin, reflects gonadal function.

Purification and quantification of lactoferrin in equine seminal plasma.

Lactoferrin with a molecular mass of 80 kDa was purified from equine seminal plasma by heparin-Agarose affinity chromatography and Sephacryl S-200 gel filtration. Purified lactoferrin was found to be highly homogeneous on the bases of its migration as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and of the monospecificity of rabbit antibodies to the purified protein in immunoblotting of seminal plasma proteins. A sandwich enzyme-linked immunosorbent assay was developed for quantifying lactoferrin in equine seminal plasma. Seminal plasma lactoferrin concentrations in 23 normal stallions ranged from 42 to 453 microg/ml, with a mean value of 157 +/- 118 microg/ml (S.D.).