Cystic ovarian disease in dairy cattle: Diagnostic accuracy when using B-mode and color Doppler ultrasound.

The most frequently reported definition of cystic ovarian disease in cattle is an abnormally persistent follicle (>7 to 10 d) with a diameter >25 mm. Discrimination between luteal and follicular ovarian cystic structures has traditionally been conducted by measuring the rim width of luteal tissue. The most common practice used in the field for diagnosis of cystic ovarian disease is examination by rectal palpation with or without the use of a B-mode ultrasound. Color Doppler ultrasound technology allows assessment of blood flow area measurements in the ovary, which has been proposed as a potential indirect measure for plasma progesterone (P4) concentrations. The objective of this study was to compare the diagnostic accuracy of differentiating luteal structures from follicular ovarian cysts using measures collected with B-mode and color Doppler transrectal ultrasonography. The definition of an ovarian cyst was a follicle greater than 20 mm in diameter in the absence of a corpus luteum that persisted for at least 10 d. A 3-mm luteal rim width was used to differentiate follicular and luteal cysts. A total of 36 cows were enrolled in the study during routine herd reproductive examination visits, with 26 and 10 having follicular and luteal cysts, respectively. Cows enrolled in the study were examined using a Mini-ExaPad mini ultrasound with color Doppler capabilities (IMV Imaging Ltd.). Blood samples were collected from each cow to measure P4 serum concentrations. History and signalment of each cow, including days in milk, lactation, times bred, days since last heat, milk composition, and somatic cell counts, were retrieved from an online database (DairyComp 305, Valley Agricultural Software). The accuracy of diagnosing follicular from luteal cysts based on luteal rim thickness was analyzed by receiver operating characteristic (ROC) curve using P4 as the gold standard, where P4 concentrations exceeding 1 ng/mL was defined as luteal, and all other structures with less P4 were considered follicular. Luteal rim and blood flow area were selected for further analysis because they presented the best ROC curves for differentiating cystic ovarian structures, with areas under the curve of 0.80 and 0.76, respectively. Luteal rim width of 3 mm was used as the cutoff standard in the study, resulting in sensitivity and specificity of 50% and 86%, respectively. Blood flow area of 0.19 cm2 was used as the cutoff standard in the study, resulting in sensitivity and specificity of 79% and 86%, respectively. When combining the use of luteal rim width and blood flow area to differentiate cystic ovarian structures, a parallel approach resulted in sensitivity and specificity of 73% and 93%, respectively, whereas an in-series approach resulted in sensitivity and specificity of 35% and 100%, respectively. In conclusion, the use of color Doppler ultrasonography when discriminating between luteal and follicular ovarian cysts in dairy cattle resulted in higher diagnostic accuracy compared with using B-mode ultrasonography alone.

B-mode and color Doppler ultrasonography of normal external jugular vein in donkeys (Equus asinus).

BACKGROUND: Although the jugular vein is a major important blood vessel in equine, the literature lacks this vessel's normal B-mode and Doppler ultrasonographic examinations in donkeys. Therefore, this study aimed to determine the reference ranges of B-mode and Doppler ultrasonographic indices of jugular veins in healthy adult donkeys (Equus asinus) and the possible effect of examination side (left and right), gender, and body condition on the ultrasonographic measurements of this vessel. B-mode and Doppler ultrasound imaging of the external jugular vein was conducted on 20 adult healthy donkeys of both sexes. RESULTS: In all donkeys, the jugular vein was 4.01 to 8.1 mm from the body surface. The longitudinal and transverse venous diameters ranged from 3.94 to 10.5 mm and from 0.88 to 1.9 cm, respectively. Moreover, the vein areas varied from 0.61 to 2.83 cm2. The reference values of superficial and deep wall thickness (SWT and DWT) were 0.56 ± 0.2 and 0.6 ± 0.13 mm, respectively. The blood velocity, blood follow rate, and congestion index of the external jugular vein can be expected in adult healthy donkeys as a range value from 8.4 to 13.5 cm/sec, from 0.33 to 1.78 ml/min, and from 0.06 to 0.27 cm.sec, respectively. Generally, the vein showed laminar monophasic waveforms. The examination side and gender have no significant effect on the ultrasound measurements of the vein (P > 0.05). Donkeys with a body condition score (BCS) ≥ 3 revealed increases in the depth of the vein (P < 0.05). CONCLUSIONS: The results of this study can be used as reference values and provide a basis for comparison when evaluating donkeys with diseases that affect blood flow in the external jugular vein.

Detection of nonpregnant cows and potential embryo losses by color Doppler ultrasound and interferon-stimulated gene expression in grazing dairy cows.

Many studies have been conducted to estimate pregnancy losses between 19 and 34 d after artificial insemination (AI) in dairy cows managed under confinement-based systems, but few studies have examined embryo mortality during this interval in dairy cows managed under gazing systems. The objectives of this prospective cohort study were (1) to assess the diagnostic value of the corpus luteum (CL) blood perfusion (BP) evaluation by Doppler ultrasound (US) to detect nonpregnant cows at 19 to 20 d post-AI, and (2) to assess the rate of potential embryo mortality between 19 to 34 d post-AI. The CL-BP of all cows included in the study (n = 131) was examined on farm by power and color mode of Doppler US and later using an image processing software by a second evaluator. The endometrium thickness and echotexture were evaluated by B-mode US at the same visit to assess if the nonpregnancy diagnosis could be improved at 19 to 20 d post-AI by this additional diagnostic tool. Blood samples were obtained at 19 to 20 d post-AI for progesterone (P4) measurement by chemiluminescence and to determine the mRNA expression of ISG by real-time PCR. Pregnancy diagnosis based on embryo visualization was performed at 33 to 34 d post-AI by US B-mode. In parallel interpretation, ISG15 and MX2 mRNA expression in leukocytes [sensitivity (Se), 100%] were regarded as suitable biomarkers for early pregnancy and were selected for molecular characterization of pregnancy at 19 to 20 d post-AI. At 19 to 20 d post-AI, 61.1% of the cows had positive CL-BP by Doppler US (Se, 98.0%), 62.7% had ISG mRNA expression in leukocytes over the cutoff point (Se, 95.7%), and 50.8% were positive, based on the combination of ISG mRNA expression, CL-BP by Doppler US, and P4 concentration (Se, 100%), and were considered as possible pregnant. At 33 to 34 d, the pregnancy rate was 37.4% diagnosed by the B-mode US. Based on the expression of the selected biomarkers in cows with active CL, we found that 28.1% of the cows could have potentially lost their pregnancy between 19 and 34 d post-AI. The Doppler US color mode showed similar accuracy and a higher negative predictive value than the genes selected as biomarkers. The additional B-mode ultrasound evaluation of the uterine stratum vasculare and the endometrium thickness improved the diagnostic accuracy. Therefore, assessing the CL-BP by Doppler US allowed early detection of nonpregnant cows at 19 to 20 d post-AI. The combination of early CL-BP by Doppler US (d 19 to 20) with early embryo detection by B-mode US (d 33-34) could be used to facilitate earlier rebreeding of dairy cows.

Exogenous L-arginine administration improves uterine vascular perfusion, uteroplacental thickness, steroid concentrations and nitric oxide levels in pregnant buffaloes under subtropical conditions.

Heat stress (HS) during pregnancy adversely affects uterine vascular perfusion and foetal development. L-arginine (L-Arg), a nitric oxide (NO) precursor, has been proven to enhance an organ's vascular perfusion. Therefore, this study aimed to examine the effect of L-Arg administration on the pregnant buffaloes' uterine haemodynamics and uteroplacental thickness under environmental HS conditions. For this purpose, pluriparous (n = 12) HS-pregnant buffaloes (mid-gestation, 180-190 days; temperature humidity index >85) were haphazardly assigned into two groups, either administered a single intravenous (IV) bolus of 5 mg/kg BW of L-Arg Hcl (n = 6; ARG) or received normal saline (0.9%; IV; 25 ml) that served as a control group (n = 6; CON). Combined uteroplacental thickness (CUPT, mm), middle uterine arteries' (MUA) diameter (mm) and uterine Doppler haemodynamics (ipsilateral and contralateral to the pregnant horn) were examined utilizing B-mode and colour Doppler ultrasonography, respectively. The serum levels of progesterone (P4), estradiol 17ß (E2), nitric oxide (NO) and total antioxidant capacity (TAC) were assayed using commercial kits. The experimental time points were - 1, 0, 2, 4, 24, 48, 72, 96 and 120 h post-L-Arg administration. The ARG group showed higher (p < .05) CUPT and MUA diameter values starting at 24 h post-treatment and onward than the control buffalo cows. In addition, there were improvements (p < .05) in the blood flow parameters in the ipsilateral MUA after L-Arg treatment evidenced by lower values of pulsatility and resistive indices starting at 48 and 4 h, respectively and onward, and higher values of peak systolic velocity, coloured areas towards the pregnant uterine horn (both 24-72 h). Increases in the NO levels were found during the period between 4 and 120 h in the ARG group compared to the CON group. Moreover, significant increases in the E2 and P4 means were noted in the ARG group, especially at 24 h onward, respectively compared to the CON group. Concerning the TAC status, neither the administration nor the hours affected serum TAC levels in the HS pregnant buffaloes (CON and ARG). In conclusion, L-Arg administration improved uteroplacental thickness and enhanced uterine haemodynamics, NO levels, and steroids production in mid-pregnant buffalo cows under environmental heat stress conditions which could improve foetal growth and development.

Ultrasonography of sudden swollen tongue in a calf.

BACKGROUND: In cattle, the lingual diseases are primarily diagnosed postmortem by histopathological examination of the affected tongues obtained after the death or during necropsy. In humans, ultrasonography has been used to provide differential diagnoses, and for preoperative or intraoperative planning of glossectomy in various lingual diseases. This is a bovine clinical case report, in which ultrasonography for sudden swelling of the tongue, which was possibly caused by snake bite, was utilized as a preoperative indication to perform a glossectomy. CASE PRESENTATION: An eight-month-old female Japanese black calf presented with sudden swelling of the tongue with well-defined discoloration in the cranial region. A 10-MHz linear probe on a portable-type ultrasound machine (MyLabOne VET, Esaote Co., Genova, Italy) was applied to the ventral surface of the tongues in the affected case, and also in five healthy calves under sedation to observe normal tongues. Ultrasonography of the swollen tongue in this case revealed that the ventral lingual muscular layers were severely thickened compared with those of normal tongues. However, the muscle layers were regularly aligned with the echogenic muscular fibers. This resembled the lingual muscular architectures of normal tongues. Color-flow Doppler ultrasonography revealed that blood flow was weakened in the small peripheral vessels in the spaces between the lingual muscular structures, and was lacking in the deep lingual artery between the apex and base of the tongue. This finding was very different than that of normal tongues, which exhibited weakened or rich blood flows. Based on ultrasonographic findings, this case was treated with glossectomy. After recovery, the calf grew up normally with a normal appetite and rumination, and did not exhibit mouth pain behavior. Histopathologically, hemorrhagic necrotic changes, together with focal formation of fibrin thrombus in the lingual blood vessels in the affected tongue, were noted. CONCLUSIONS: To the best of our knowledge, the present report is the first description of lingual ultrasonography performed in cattle. In this case, ultrasonography enabled visualization of decreased vascularity, which might be associated with hemorrhage or formation of fibrin thrombus in the suddenly swollen tongue presented.

Short communication: Accuracy of corpus luteum color flow Doppler ultrasonography to diagnose nonpregnancy in dairy cows on day 21 after insemination.

The main objective of this study was to determine the accuracy of corpus luteum color flow Doppler ultrasonography (CLCFDU) to diagnose nonpregnant dairy cows 21 d after insemination. A secondary objective was to determine the accuracy of other indirect tests such as corpus luteum size and progesteronemia. Data from 1,632 Holstein cows (10 commercial herds) were used for the analysis in this prospective cohort study. The herds were visited weekly by a veterinarian and an animal health technician. During farm visits, cows were examined on d 21 after insemination using transrectal B-mode ultrasonography to quantify the presence and size of the corpus luteum. After identification of the corpus luteum, CLCFDU was performed and scored as D0, D1, D2, or D3 when 10% or less, between 11 and 30%, between 31 and 60%, or 61% or more of the corpus luteum surface was colored, respectively. A blood sample from coccygeal vessels was also collected to quantify progesteronemia. Farmers were blinded to these findings and no intervention was performed following examination. On d 32 after insemination, the cows were examined by the regular herd veterinarian using transrectal palpation and B-mode ultrasonography to determine whether the cows were pregnant or not (the reference test). Statistical analyses were conducted using 2 × 2 contingency tables. The apparent prevalence, sensitivity, specificity, positive predictive value, and negative predictive value of CLCFDU for predicting nonpregnancy were 22.0, 36.6, 99.0, 98.1, and 52.0%, respectively, when using D0 only as the diagnostic criterion; they were 47.2, 76.4, 94.8, 93.5, and 73.8%, respectively, for D0+D1 criteria. The same measures for cows with a corpus luteum <15 mm were 8.0, 11.7, 97.5, 86.9, and 43.4%, respectively, and they were 51.9, 67.4, 70.4, 76.6, and 60.0%, respectively, for progesteronemia <1 ng/mL. The measures of accuracy of CLCFDU to identify nonpregnant cows on d 21 after insemination were high, and the apparent prevalence varied depending on the diagnostic criteria used. The measures of accuracy of corpus luteum size (<15 mm) for the same purpose were high; however, apparent prevalence was low. The measures of accuracy of progesteronemia (<1 ng/mL) were low. In conclusion, CLCFDU had excellent accuracy to diagnose nonpregnancy in dairy cows on d 21 after insemination.

Ultrasonographic evaluation of umbilical structures in Holstein calves: A comparison between healthy calves and calves affected by umbilical disorders.

The aim of this study was to evaluate, during the first 4 wk of life, the involution of umbilical structures in clinically healthy calves and in calves affected by umbilical disorders, in both B-mode and color Doppler ultrasonography. The present study was carried out on 63 Holstein Friesian calves, divided into 3 different groups: group H (clinically healthy, n = 17), group A, (affected by omphaloarteritis, n = 24), and group V (affected by omphalophlebitis, n = 22). B-mode ultrasonography was performed at weekly intervals, using a portable device (LOGIQ Book XP, GE Healthcare, Little Chalfont, UK) and a linear multifrequency 7 to 10 MHz probe. In addition to the ultrasound examinations, the umbilicus of the calves was also evaluated using a color Doppler. The extra-abdominal as well as the intra-abdominal hemodynamics within the umbilical structures were evaluated using 3 different ranges of speed-flow detections (23, 14, and 7 cm/s). In healthy calves, as the age increased, the umbilical structures decreased in size and their ultrasonographic identification became more difficult. Conversely, in affected calves the umbilical structures did not show the same progressive reduction of external diameters and areas, but had an irregular trend of regression. Also the color Doppler ultrasonography showed a significant difference in frequency and percentage of speed-flow in the early days of examination between the 3 groups, with the most reliable results detected from the umbilical arteries. In our experimental study, omphaloarteritis could be detected at the d 1 color Doppler exam, with a 100% specificity and a 100% positive predictive value.

Doppler sonography of uterine blood flow at mid-oestrus during different degree of clinical endometritis in dairy cows.

The present study was aimed at analysing the mid-oestrous uterine blood flow parameters during varying degree of endometritis in dairy cows. Degree of clinical endometritis was adjudged on the basis of visual examination of uterine discharge collected from cows at mid-oestrus, that is mild (slightly turbid; n = 13), moderate (turbid with pus flakes; n = 14) and severe (milky; n = 13). Pulsatility and resistance indices (PI and RI), time average mean velocity (TAMEAN), time average maximum velocity (TAMAX), diameter of the artery, volume of blood flow, Doppler pulse duration (DPD) and systolic upstroke/acceleration time (AT) were measured to study the spectral waveforms at mid-oestrus. Significantly higher (p < .01) PI and RI were found in cows diagnosed with mild degree in comparison with cows diagnosed with moderate and severe degrees of clinical endometritis. There was significantly higher (p < .01) velocity (TAMEAN and TAMAX), volume of blood flow (BFV-TAMEAN, BFV-TAMAX) and DPD in both the middle uterine arteries during moderate and severe degrees of clinical endometritis as compared to mild endometritis. However, significantly higher (p < .01) AT was recorded in cows diagnosed with mild degree as compared to moderate and severe degrees of clinical endometritis. Pearson's correlation analysis has shown that RI was positively correlated with PI and AT in all the groups under study (mild degree, r = .72 and .49; moderate degree, r = .54 and .38; severe degree, r = .90 and .42; p < .05). However, there found significantly negative correlation (p < .05) with other parameters in all the cows irrespective of degree of inflammation. Therefore, it may be deducted that assessment of uterine inflammation can be done with a non-invasive technique known as Doppler sonography which can be useful in adjudging the hemodynamic changes inside uterus and future fertility of dairy cows.

Colour Doppler ultrasound imaging of blood flows variations in neoplastic and non-neoplastic testicular lesions in dogs.

Testicular tumours are the most common neoplasms in male dogs accounting for approximately 90% of all tumours affecting the genitourinary tract. Gray-scale ultrasonography in combination with colour and power Doppler imaging has been well accepted as an accurate technique for assessing scrotal lesions and vascularization of the testis. Colour Doppler sensitivity for low blood flows appears promising in the study of testicular disorders. The aim of this study was to assess if colour and power Doppler ultrasound is a good tool for the investigation of testicular lesions in dogs, to report the sonographic features of lesions and to measure colour and power Doppler parameters such as resistive index (RI), pulsatility index (PI), hypovascularization and hypervascularization (VI) determining if they can be used to distinguish testicular neoplasms from the wide spectrum of non-neoplastic pathological findings. In this study, 50 male dogs of various breeds, aged between 7 and 14 years, presented with testicular disorders were selected. RI and PI were calculated. Mean RI values for neoplastic, inflammatory and degenerative lesions were 0.54, 0.45 and 0.58, respectively. Mean PI values were 0.62, 0.55 and 0.63, respectively. Hypovascularization and hypervascularization of the lesion were evaluated throughout the vascularity index (VI). Vascular signals in neoplasms were significantly intensified around and inside the mass if compared with those measured during inflammatory and degenerative lesions. VI markedly increased in solid tumours. Pathological testes were removed; macroscopical, histological and immunoistochemical evaluations were carried out. Colour Doppler showed increased intralesional and peripheral flows in all neoplastic lesions analysed. No flows were detected around cysts.

The relationship between mastitis and the B-mode, colour Doppler ultrasonography measurements of supramammary lymph nodes in cows.

This research communication aims to test the hypothesis that B-Mode, colour Doppler ultrasonographic measurements and characteristics can identify mastitis in dairy cows. A total of 102 lactating cows were divided into 3 groups: cows in which all mammary quarters were CMT-negative, cows with CMT-positive mammary quarters and cows with clinical mastitis in at least one quarter. Colour Doppler ultrasonography measurements of the supramammary lymph nodes revealed that distortion-type vascular morphology, the rate of type 4 vascular densities and the incidence of mixed-type vascular distributions were highest in the clinical mastitis group, whereas the frequency of avascularity in supramammary lymph nodes was highest in the CMT-negative group. All differences were significant. In conclusion, the use of B-mode and colour Doppler ultrasonographic measurements of the supramammary lymph nodes can provide useful information about the current condition of mastitis in cows, although its diagnostic potential remains to be determined.

Vascular ultrasonographic findings in canine patients with clinically diagnosed phlebitis.

Peripheral vein phlebitis (inflammation) is a relatively frequent complication in dogs, however, published information on the ultrasonographic characteristics is currently lacking. This prospective, observational study describes the ultrasound (US) characteristics of normal canine cephalic veins, and veins with clinical phlebitis. Correlations among US findings and between US findings versus time that the intravenous catheter was in place were investigated. Safety of the US procedure was evaluated. Fifty patients were prospectively recruited for the study and 18 met the final inclusion criteria. Each patient underwent daily US examinations and was assessed for multiple criteria (vascular wall appearance, compressibility, spontaneity of flow, color fill, and presence/absence of filling defects, flow contour, direction, non-pulsatility). Characteristics of normal canine cephalic veins were as follows: smooth and thin wall, complete compressibility, no flow disturbances, no filling defects, smooth flow contours, and unidirectional, non-pulsatile flow with no turbulence. Characteristics of cephalic veins with clinical phlebitis were as follows: wall thickening (83%), decreased compressibility (55%), filling defects consistent with intraluminal thrombus (55%), vessel wall hyperechogenicity (44%), and abnormal color Doppler flow (39%). Significant correlations were found between Doppler filling defects and compressibility, Doppler filling defects and presumed thrombosis, and compressibility and presumed thrombosis (P = .001, P = .001, P = .000, respectively). No correlation was found between the US findings and time the intravenous catheter was in place. Findings indicated that duplex and compressibility US are feasible and safe methods for characterizing and monitoring cephalic veins in dogs with clinical phlebitis.

Colour-Doppler ultrasound imaging as a laparoscopy substitute to count corpora lutea in superovulated sheep.

This study evaluated colour-Doppler ultrasound imaging (UI) as a substitute for laparoscopy to count the corpora lutea (CL) in superovulated sheep. Twenty-five Santa Ines ewes were superovulated three times at 21-day intervals. Corpora lutea were counted by colour-Doppler UI (CLDOPPLER ) 6 days after each superovulation and confirmed by laparoscopy (CLLAP ) 12 hr later. The mean number of CL was similar for both techniques (2.1 ± 2.5 vs. 2.1 ± 2.7 for CLDOPPLER and CLLAP , respectively) with a significant positive correlation (r = .94; r2 =.89). Colour-Doppler UI effectively evaluated the ovarian response in superovulated ewes and efficiently identified animals that did not respond to superovulation.

Mineralization can be an incidental ultrasonographic finding in equine tendons and ligaments.

Tendon/ligament mineralization is recognized in horses but information regarding its clinical significance is limited. The aims of this observational study were to report the structures most commonly affected by ultrasonographically detectable mineralization and, for these, determine frequency of diagnosis and key clinical features. Cases presented at our hospital in April 1999-April 2013 and September 2014-November 2015 were included: a total of 27 horses (22 retrospective, five prospective). Mineralizations were most common in deep digital flexor tendons (10) and suspensory ligament branches (eight), representing 10% and 7% (estimated), respectively, of horses diagnosed with injuries to these structures during the study. Two deep digital flexor tendon and three suspensory ligament branch cases showed bilateral mineralization. Deep digital flexor tendon mineralization was restricted to the digital flexor tendon sheath, most commonly in the proximal sheath (±sesamoidean canal), and seven of 10 cases involved hindlimbs. Suspensory ligament branch mineralization was visible in the same ultrasound window as the proximal sesamoid bones in 10/11 limbs and six of eight cases involved forelimbs. Previous corticosteroid medication was a feature of one deep digital flexor tendon and one suspensory ligament branch case. Mineralization was associated with lameness in some but not all limbs. Mineralized foci within the deep digital flexor tendon preceded hypoechoic lesion formation in two limbs. Of the cases with deep digital flexor tendon or suspensory ligament branch injury only, one of three and two of three cases, respectively, became sound. Findings indicated that tendon/ligament mineralization can be associated with lameness in some horses, but can also be an incidental finding.

Renal evaluation with B-mode and Doppler ultrasound in captive tufted capuchins (Sapajus apella).

BACKGROUND: This study aimed to evaluate, by B-mode ultrasound and Doppler ultrasound, the kidneys of one Neotropical primate maintained in captivity. METHODS: Ten adult intact tufted capuchins (Sapajus apella) were evaluated. The morphometric parameters of both kidneys were evaluated and kidney volumes calculated. Color Doppler was used to evaluate renal vascular architecture and to identify arteries of the left kidney. Spectral Doppler was used to measure peak-systolic velocity, end-diastolic velocity, and resistive index (RI). RESULTS: Both kidneys presented homogeneous cortical echogenicity and homogeneous echotexture with preserved corticomedullary differentiation. There was no statistically significant difference between right and left kidneys for all morphometric parameters. RI of the left renal, interlobar, and arcuate arteries were, respectively, 0.75 (SD, 0.07), 0.70 (SD, 0.08), and 0.70 (SD, 0.06). CONCLUSIONS: Both kidneys showed similar morphometric parameters by B-mode ultrasound, whereas RI by spectral Doppler decreased progressively from renal artery to interlobar and arcuate arteries.

Ovulation of the preovulatory follicle originating from the first-wave dominant follicle leads to formation of an active corpus luteum.

The objective of our study was to compare the characteristics of the corpus luteum (CL) formed after ovulation of the dominant follicle (DF) of the first follicular wave (W1) and those of the CL formed after ovulation of the DF of the second (induced) follicular wave (W2). Non-lactating Holstein cows were used for this study. In Experiment 1, cows were treated with PGF2α and GnRH on days 6 and 8 (day 0 = day of follicular wave emergence) for W1 (n = 6) and W2 (n = 6), respectively. Dominant follicles were aspirated on day 9 to quantify the amounts of mRNA (VEGF120, VEGF164, FGF-2, StAR, P450-scc and 3ß-HSD) in granulosa cells (GC). In Experiment 2, the size and blood flow area of the CL formed after ovulation of the DF in W1 (W1CL; n = 6) and W2 (W2CL; n = 6) (the day of DF ovulation in W1 and W2 was day 10) were evaluated on days 12, 15, 18 and 21. The plasma P4 concentration was measured on days 10 to 21. The amounts of VEGF164, P450-scc and 3ß-HSD mRNA were higher (P < 0.05) in the DF in W1, and those of VEGF120,FGF-2 and StAR mRNA tended to be higher (P < 0.1) in the DF in W1. The size of the CL was greater in the W1CL on days 15, 18 and 21. The blood flow area of the CL was greater in the W1CL on days 12 and 15. The plasma P4 concentrations were higher in the W1CL. These results indicate that the CL formed after ovulation of the DF in W1 was greater in terms of size, blood flow and plasma P4 concentration.

Colour Doppler Ultrasonography as a Tool to Assess Luteal Function in Santa Inês Ewes.

The aim of this study was to evaluate luteal dynamics in the Santa Inês ewes using colour Doppler (CD) ultrasonography. Oestrus was synchronized in nulliparous females (n = 18), and subsequently, they were only teased (n = 6) or teased and mated (n = 12). Blood samples were collected daily for plasma progesterone (P4 ) concentrations. Ultrasonographic images of corpora lutea (CL) in CD mode were obtained for further analysis in its largest diameter. The CD mode allowed an early sequential monitoring of CL that was visualized by the first time 0.77 ± 0.62 days after ovulation, with luteal area 29.68 ± 13.21 mm(2) . During the luteogenesis, a progressive increase was observed, followed by a plateau of luteal area, vascularization area and plasma concentrations of P4 reaching maximum values in D11 (124.0 ± 38.0 mm(2) , 52.78 ± 24.08 mm(2) and 11.23 ± 4.89 ng/ml, respectively). In the luteolysis, the plasma concentrations of P4 decreased sharply, whereas luteal and vascularization area gradually. The vascularization area was positively correlated with plasma concentrations of P4 during the luteogenesis (r = 0.22) and luteolysis (r = 0.48). The luteal dynamics of Santa Inês ewes showed patterns similar to those observed in other sheep breeds studied. The CD ultrasonography has the potential to be used as a tool to assess luteal function in sheep.

Imaging diagnosis--seminoma causing liver compression in a spur-thighed tortoise (Testudo graeca).

A 13-year-old male spur-thighed tortoise (Testudo graeca) was presented with anorexia, apathy, and prolapse of penile tissue. Ultrasonography revealed a large heterogeneous mass in the coelomic cavity, and fine-needle aspiration demonstrated sperm. Magnetic resonance imaging showed a sharply defined mass originating from the left testis. Appearance and signal intensities were similar to those reported in testicular neoplasms in humans, in particular sharing similarities with seminomas. Necropsy results and histopathological findings were consistent with a seminoma. To the authors' knowledge this is the first report of the diagnosis of testicular neoplasia in a reptile using imaging techniques.

Conceptus-induced changes in the gene expression of blood immune cells and the ultrasound-accessed luteal function in beef cattle: how early can we detect pregnancy?

We aimed to identify the functional characteristics of the corpus luteum (CL) by color Doppler ultrasonography and changes in interferon-stimulated gene (ISG) expression in peripheral blood mononuclear cells (PBMCs) during early pregnancy in beef cows. We then aimed to use these features to establish earlier pregnancy diagnosis methods. In experiment 1, the CL size and blood flow were accessed by Doppler ultrasonography, and the PBMCs were isolated on Days 8, 12, 15, 18, 20, 22, 25, 30, 45, and 60 post-timed artificial insemination (TAI) from pregnant (n = 10) and nonpregnant cows (n = 12). The abundance of ISG (OAS1, MX1, MX2, and ISG15) transcripts was measured by quantitative PCR. Analyses of OAS1 and MX2 expression in isolated PBMCs (ISG-PBMC method) and Doppler imaging of CL (Doppler-US method) were performed to test the accuracy of these methods for the diagnosis of pregnancy on Day 20 post-TAI (n = 110; experiment 2). In experiment 1, the luteal volume and blood flow were reduced in nonpregnant cows during the first weeks post-TAI, but an evaluation of CL vascularization and size was efficient in identifying nonpregnant cows on Day 20 post-TAI. The expression of ISGs in PBMCs can be stimulated by the presence of a viable conceptus between Days 15 and 22 post-TAI, and the expression of these genes reaches a peak on Day 20. In experiment 2, the Doppler-US and ISG-PBMC methods resulted in similar specificity (85.5 and 87.7%, respectively). However, only the Doppler-US method resulted in 100% sensitivity. In conclusion, the greatest abundance of ISGs in PBMCs and a high detection of luteolysis by Doppler imaging on Day 20 post-TAI can be feasibly used for the earlier detection of nonpregnant cows in reproductive programs. The level of accuracy for our described pregnancy methods is high on Day 20 (80%-91%), but only the Doppler imaging method results in an absence of false-negative diagnoses.