Effect of follicular aspiration at the onset of progesterone-based timed artificial insemination on the follicular dynamics and fertility of early postpartum Japanese black cows

The effect of gonadotropin-releasing hormone analogue (GnRH-A) or follicular aspiration at the onset of progesterone-based timed artificial insemination (TAI) on subsequent follicular growth and synchronization of ovulation was examined in early postpartum Japanese Black cows. A total of 40 (22 in Exp. 1 and 18 in Exp. 2) Japanese Black cows at 20-30 days postpartum were fitted with a progesterone releasing internal device (PRID) for 7 days, injected with a prostaglandin F2α analogue upon removal of the PRID and GnRH-A 48 h later, and inseminated 18 h after GnRH-A injection. In Exp. 1, the animals were divided into three groups (untreated control, GnRH-A injection or follicular aspiration) of different treatments on the first day of PRID insertion (day 0), and the synchronized ovulation rate in the follicular aspiration group (100%; 8/8) tended to be higher (P = 0.077) than that in the control group (42.9%; 3/7). In Exp. 2, follicular growth in the GnRH (n = 9) and follicular aspiration (n = 9) groups was monitored by ultrasonography. Four out of the nine animals in the GnRH group had a corpus luteum on either day 4 or day 7 (OV group), and the other five animals had no induced ovulation (NOV group). The diameter of the ovulatory follicle on day 9 in the OV group (1.44 ± 0.11 cm) tended to be greater (P = 0.078) than that in the NOV group (1.13 ± 0.07 cm). Follicular aspiration at the onset of PRID-based TAI of early postpartum Japanese Black cows, regardless of the resumption of ovarian cyclicity, tended to result in a higher rate of synchronization of ovulation than that of the untreated controls.     

Conversion of chromosome-specific RAPDs into SCAR-based anchor markers for onion linkage maps and its application to genetic analyses in other Allium species

Integration of previously developed Allium cepa linkage maps requires the availability of anchor markers for each of the eight chromosomes of shallot (A. cepa L. common group Aggregatum). To this end, eight RAPD markers originating from our previous research were converted into SCAR markers via cloning and sequencing of RAPD amplicons and designing of 24-mer oligonucleotide primers. Of the eight pairs of SCAR primers, seven resulted in the amplification of single bands of the original RAPDs, and the remaining primer set amplified an additional band. The results of Southern hybridization using RAPD amplicons from genomic DNA of Japanese bunching onion (Allium fistulosum L.)—shallot monosomic addition lines indicated that five SCAR markers were single shallot chromosome-specific markers and were not detected in genomic DNA of A. fistulosum. The eight SCAR primer pairs were applied to other Allium species and exhibited three types of amplification profiles, namely RAPD amplicons observed only in shallot, in shallot and Allium vavilovii, and in several Allium species. A mapping study using 65 F₂ plants generated by the selfing of one interspecific cross A. cepa × Allium roylei individual integrated the SCAR marker SAOE17₅₀₀ into chromosome 5 as expected. The results of the present study show that the eight SCAR primer sets specific to shallot can facilitate the mapping in A. cepa and can also serve as anchor points between maps of different Allium species.     

Diagnostic imaging of intra-abdominal cyst in heifer using the computed tomography

A 10-month-old Japanese black heifer was diagnosed as having an intra-abdominal cyst using computed tomography (CT). Through a posterior ventral midline incision, the cyst was removed, and the heifer completely recovered after the surgery. CT scans enabled detection of the intra-abdominal cyst and measurements of the diameter of the cyst before the surgery.     

Steroid injection therapy in a feline solitary bone cyst

A nine months old Japanese domestic cat with a solitary bone cyst, which is a benign tumor-like lesion and is particularly uncommon in feline practice was referred. Radiographic examination revealed an expansive radiolucency in the distal metaphysis of the right ulna and pathologic fracture. The histological examination demonstrated immature osteogenesis consisting of osteoblasts surrounded by connective tissue. We applied drainage and instillation of steroid solution into the cystic cavity. Clinical signs, including lameness and pain, disappeared three weeks after the therapy started. Fourteen months after the therapy, the cystic lesion of bone was remodeling successfully without re-developing. We conclude that our procedure was useful treatment in this case.     

Plasma concentration of estrone sulfate during pregnancy in different breeds of Japanese beef cattle

Plasma concentrations of estrone sulfate in different breeds of Japanese beef cattle and the relationship between those concentrations and feto-placental growth were examined in order to assess the possibility of monitoring abnormal growth of the fetus. Blood samples were obtained from cows from day 90 of gestation to parturition. The plasma concentration of estrone sulfate was measured by direct enzyme immunoassay. From day 180 of gestation, the mean concentration of estrone sulfate increased gradually and it was drastically elevated after day 240 of gestation with the maximum at day 285. Plasma concentrations of estrone sulfate on day 240 of gestation was significantly increased in F(1) cows (Holstein Friesian and Japanese Black) compared with those in other breeds of cow. From day 270 to 278 of gestation, estrone sulfate concentrations of Holstein Friesian cows inseminated by Holstein Friesian differed from those inseminated by Japanese Black. In the cow with retained placenta, the plasma concentration of estrone sulfate reached plateau at day 240 of gestation and did not increase thereafter. There was no significant relationship between estrone sulfate concentration and duration of gestation, calf birth weight, weight of placenta or viability of newborn calves. These results indicate that changes of plasma estrone sulfate concentration in Japanese beef cattle are very similar to those in Holstein dairy cattle. They also suggest that the plasma concentration of estrone sulfate is associated with the breed of pregnant cow and that its concentration is also affected by calf birth weight depending on the breed of bull. It seems possible to predict the incidence of retained placenta but not the calf birth weight and viability of newborn calves in Japanese beef cattle.     

Three-dimensional image analysis of a head of the giant panda by the cone-beam type CT

The cone-beam type CT (Computed Tomography) enabled us to collect the three-dimensional (3D) digitalized data directly from the animal carcass. In this study, we applied the techniques of the cone-beam type CT for a carcass head of the giant panda (Ailuropoda melanoleuca) to obtain the 3D images easily without reconstruction process, and could morphologically examine the sections from the 3D data by means of non-destructive observations. The important results of the study represent the two following points. 1) We could show the morphological relationships between the muscles of mastication and the mandible in non-destructive status from the 3D data. The exact position of the coronoid process could be recognized in the rostro-lateral space of the temporal fossa. 2) By the serial sections from the 3D data sets, the morphological characteristics in the nasal cavity were detailed with high resolution in this rare species. The nasal concha was well-developed in the nasal cavity. The ethmoidal labyrinth was encountered immediately caudal to the nasal cavity and close to the region of the olfactory bulb. The ethmoidal labyrinth consisted of the complicated osseous structure in this area. The data will be useful to discuss the olfactory function in the reproduction behavior of this species.     

Concentration patterns of antibacterial factors and immunoglobulin A antibody in foremilk fractions of healthy cows

Antibacterial factors act as innate immune components, which respond as soon as bacteria enter a living organism. To prevent and treat mastitis in cattle, understanding the concentrations of these substances inside the udder is important; however, they remain to be studied. In this investigation, the concentration of lingual antimicrobial peptide (LAP), S100 protein (S100A7), lactoferrin (LF), and immunoglobulin antibody were measured in the different fractions of foremilk. Lactating Holstein cows were examined, and 10 foremilk fractions were obtained from sequential samples up to 150 ml. The LAP concentrations in milk samples increased until 25 ml. The LF concentrations increased up to the 10 ml fraction, then stabilized at low level after the 50 ml fraction. For S100A7, some fractions had significantly higher (p < .05) concentrations than the 5 or 10 ml fractions. The IgA antibody concentration increased up to the 5 ml fraction, then after 50 ml fraction showed relatively low concentrations. This investigation determined the concentration patterns of LAP, LF, S100A7, and IgA antibody secreted in milk inside the udders of healthy lactating cows as baseline data. These distinct concentration patterns might indicate various protective responses.     

Lingual antimicrobial peptide and lactoferrin concentrations and lactoperoxidase activity in bovine colostrum are associated with subsequent somatic cell count

The present study was undertaken to examine whether potential levels of innate immune factors (lingual antimicrobial peptide (LAP), lactoferrin (LF) and lactoperoxidase (LPO)) in colostrum are associated with subsequent milk somatic cell count (SCC) in dairy cows. Quarter milk samples were collected daily for 1 week postpartum to measure LAP and LF concentrations and LPO activity. SCC in milk was determined weekly for 2 months postpartum and its correlations to concentrations of LAP and LF and LPO activity were examined. Only small variations of all immune factors were found among four udders in each individual cow, whereas there were great differences in these factors among cows. Negative correlation was detected only between LPO activity and mean and maximum SCC, whereas its relationship was not significant. LAP and LF concentrations were significantly correlated positively to mean, maximum and minimum SCC. These results suggest that the great difference in innate immune factors among animals and high LAP and LF concentrations in colostrum may be associated with subsequent high incidence of SCC increase.     

Comparison of cadherin and integrin localization in bovine cystic and healthy follicles

As stage progresses in the cystic follicle, granulosa cells are lost. We hypothesized that the granulosa and theca interna layers are detached in association with weakened expression of cell adhesion molecules such as cadherin (cell–cell adhesion) and integrin (cell–extracellular matrix adhesion) in cystic follicles. To elucidate this hypothesis, we immunolocalized these molecules in the granulosa and theca interna and compared them between cystic and small healthy follicles. Sections were immunostained with cadherin and integrin β₁ antibodies and their localizations were compared. Cadherin‐positive reaction was seen in the cytoplasma of all granulosa cells. No increase in the frequency of cadherin‐positive area in the granulosa layers and the intensity of cadherin immunoreaction in the theca interna was detected in cystic follicles compared with healthy ones. A dense immunoreaction product of integrin β₁ was detected in the theca interna in both cystic and healthy follicles. Intensity of integrin β₁‐immuno reaction in the granulosa layers and integrin β₁‐positive area in the theca interna was significantly lower in the cystic follicle than in the healthy follicles. These results suggest that granulosa and theca interna cells are detached while maintaining the cell–cell adhesion, resulting in the consequent loss of these layers from the cystic follicle.     

Utility of contrast-enhanced ultrasound in differential diagnosis of adrenal tumors in dogs

This prospective case study aimed to clarify the clinical significance of contrast-enhanced ultrasound (CEUS) for the differential diagnosis of canine adrenal tumors. Forty-three client-owned dogs with adrenal tumors were included. All dogs underwent CEUS, which was evaluated qualitatively and quantitatively. The peak signal intensity (PI), time to peak signal intensity (TPI), mean transit time (MTT), upslope, and downslope were calculated for each time-intensity curve. The histopathological diagnosis of each resected mass was compared with the CEUS findings and parameters. Enhancement distribution, vascularity, tortuous nourishing vessels, enhancement pattern, and late-phase enhancement did not differ significantly between adrenal cortical adenoma (CA), adenocarcinoma (CAC), and pheochromocytoma (PHEO) in qualitative assessment. In PHEO, the TPI was significantly more rapid compared with that in CA (P=0.0287) and CAC (P=0.0404). The MTT in PHEO was significantly shorter than that in CA (P=0.0016) and CAC (P=0.0003). Upslope in PHEO was larger than that in CAC (P=0.0406). Downslope in PHEO was significantly larger than that in CA (P=0.0048) and CAC (P=0.0018). A receiver operating characteristic curve analysis demonstrated that the area under the MTT curve yielded 0.91 for distinguishing PHEO from adrenocortical tumors in dogs; an MTT cut-off value less than 6,225 msec yielded a sensitivity of 69%, specificity of 94%, and likelihood ratio of 12.46. CEUS appears to be clinically applicable for the differential diagnosis between cortical and medullary origins of primary adrenal tumors in dogs.