A comparison study between the Siemens ADVIA 120 and manual method for the differential white blood cell count in goats.

BACKGROUND: Although widely used, the ADVIA 120 hematology analyzer has not been previously validated for determining the differential leukocyte count in goats. OBJECTIVES: The aim of this study was to compare the differential leukocyte counts provided by the ADVIA 120 (A-diff) and the manual method (M-Diff) in goats. METHODS: EDTA blood samples that were analyzed within 4 h of collection were used in the study. The following exclusion criteria were applied: inappropriately filled tubes or tubes containing clots, erroneous ADVIA peroxidase cytograms, and blood smears of poor quality. The A-Diff was compared with the M-Diff performed by two independent observers on 200 leukocytes. RESULTS: Forty samples were included after previously excluding eight samples. The correlation between the A-Diff and M-Diff was very strong for eosinophils (r = .870, p < .001) and strong for lymphocytes (r = .796, p < .001) and neutrophils (r = .730, p < .001), while no significant correlation was observed for monocytes (r = .026, p = .872). The Passing-Bablok regression analyses revealed statistically significant constant errors for neutrophils (5.83%; 95% confidence interval [CI]: 0.41%, 12.18%) and eosinophils (1.89%; 95% CI: 1.17%, 2.71%). Bland-Altman analyses showed a statistically significant negative bias for lymphocytes (-5.0%) and a statistically significant positive bias for eosinophils (2.2%). The very low basophil percentages precluded a meaningful method comparison. CONCLUSIONS: The ADVIA 120 overall demonstrated good performance for the differential WBC count in goats under the conditions of this study. Therefore, it can be considered suitable for routine hematologic screening in goats. Nonetheless, it should be emphasized that any abnormal result should be confirmed with a blood smear evaluation.

Udder Ultrasonography of Dairy Cows: Investigating the Relationship between Echotexture, Blood Flow, Somatic Cell Count and Milk Yield during Dry Period and Lactation.

Udder health of dairy cows is related to their productivity and welfare. The period from dry-off to calving and early lactation is crucial. Ultrasonography is a useful and practical tool for the examination of the mammary parenchyma and blood flow. This observational study investigated the relationship between udder echotexture features, blood flow volume (BFVol) in the milk vein, milk somatic cell count (SCC) and daily milk yield (DMY) from late lactation, throughout the dry period and consecutive early lactation. Seventeen repeated measurements were performed on twenty-one Holstein cows. The udder parenchyma was examined with B-mode ultrasonography. Udder echotexture was studied using 15 features: Numerical Pixel Value (NPV), Pixel Standard Deviation (PSD), Skewness, Excess, Contrast, Homogeneity, Correlation, Entropy, Run Percentage, Long-Run Emphasis, Grey Value Distribution, Runlength Distribution, Gradient Mean Value, Gradient Variance and Percentage of Non-zero Gradients. Blood flow in the milk vein was examined with spectral Doppler. Linear mixed-effects models were employed to investigate relationships between BFVol, udder echotexture features, SCC and DMY throughout the study period. Our models showed that a 1 kg increase in DMY was associated with a significant increase of 0.25 L/min in the expected BFVol and that a 1,000,000-cells/mL increase in SCC was associated with a significant BFVol decrease of 0.49 L/min, keeping all other variables constant. Multivariable models showed significant associations between DMY and NPV, between PSD and Long-Run Emphasis, and between SCC and NPV, PSD, Gradient Mean Value, Homogeneity, Gradient Variance and Entropy. In conclusion, udder echotexture and BFVol in the milk vein are related to SCC and milk yield. Ultrasonography can be used for the comprehensive assessment of udder health in support of precision dairy farming.

Association between dam and calf measurements with overall and fetopelvic dystocia in Holstein heifers.

We investigated the relationship between dam's pelvic and calf's dimensions with dystocia due to fetopelvic disproportion in the Holstein breed and estimated risk factors and dystocia probability. For this purpose, external pelvic measurements were performed in 402 heifers 15 ± 11 (1-38) days ante-partum and specific conformation measurements were obtained from their calves 1.7 ± 1.2 post-partum. Dystocia was defined as the inability of the heifer to complete parturition spontaneously within 120 min after the appearance of the amnion with normal presentation, position and posture or as having definite obstetrical obstacles within 60 min. Overall and fetopelvic disproportion dystocia incidence was 10.4% and 5.2%, respectively. Heifer measurements mainly influenced overall dystocia, whereas calf conformation was related solely with fetopelvic dystocia. Specifically, heifers with a small pelvis (hip width <49.95 cm, pelvic inlet area <333.2 cm2, pelvic volume <7799.2 cm3) had 2.8 to 3.5 times greater incidence of overall dystocia (19.0-20.8%) compared to heifers with a larger pelvis (incidence of 7.0-7.6%). Regarding calf factors, sex (male calves), body weight, chest circumference and fetlock joint circumference significantly increased the odds of experiencing dystocia due to fetopelvic disproportion compared with female, lighter or smaller calves. In a backward elimination model with independent variables treated as continuous, an area under the ROC curve of 0.66 regarding the prediction of overall dystocia based on heifer pelvic length, and of 0.64 for the prediction of fetopelvic dystocia based on fetlock joint circumference was found. The combination of the two variables in one model improved the ROC area to 0.71 regarding dystocia due to fetopelvic disproportion, reaching acceptable level of discrimination. Our findings indicate that dystocia due to fetopelvic disproportion in heifers is mainly influenced by the fetal side. Additionally, the estimation of pelvic dimensions of the dam before parturition and specific conformation characteristics of the calf during parturition, especially fetlock joint circumference, could aid obstetricians and herdsmen regarding dystocia probability and parturition surveillance.

Colour Doppler study of blood flow in the portal vein in relation to blood flow in the milk vein, milk yield and body condition of dairy cows during dry period and lactation.

In dairy cows, the liver supports the increased nutrient demands of the udder for milk production. Blood flow is key for the transport of these nutrients. This study investigated portal vein blood flow volume (PVBFVol) in relation to daily milk yield (DMY), milk vein blood flow volume (MVBFVol) and body condition parameters of high-producing dairy cows, starting from late lactation, throughout dry period, and consecutive early lactation. Seventeen repeated examinations were performed on 19 Holstein cows and 313 measurement days were finally included. Vein morphology and blood flow were examined via B-mode and spectral Doppler (triplex) ultrasonography, respectively. Body condition parameters recorded were body condition score (BCS), backfat thickness (BFT) measurement with ultrasonography, heart girth circumference (HG) and withers height (WH). Longitudinal relationship of PVBFVol with MVBFVol, DMY, BCS, BFT, HG and WH was analyzed with linear mixed models, with random intercept effects, using restricted cubic splines. A significant increase of 8.28% (p < 0.01) in PVBFVol appeared for every 1 L/min increase in MVBFVol in the univariable model. PVBFVol presented a significant negative association with BCS (p < 0.01) and BFT (p = 0.02), while interaction with production stage was significant, too. PVBFVol significantly increased by 0.38% (p = 0.04) for every 1 kg increase in DMY in the multivariable model. In conclusion, the increased PVBFVol during lactation accompanies the escalation in metabolic activity of the liver and the increased blood circulation through the udder, coping with the udder's escalating nutrient demands for milk synthesis.

The Role of Ewes' Udder Health on Echotexture and Blood Flow Changes during the Dry and Lactation Periods.

The objective of the current study was to investigate the echotextural and hemodynamic changes of ewes entering the dry period with or without subclinical mastitis. B-mode and color Doppler ultrasonography were applied to 12 Chios ewes (6 with healthy udders (group A) and 6 with subclinical mastitis (group B)) before the dry period, during the dry period (the involution phase, steady state, and transition phase), and postpartum. The color Doppler of the mammary arteries was used to evaluate them according to the pulsatility index (PI), resistive index (RI), end-diastolic velocity (EDV), time-averaged maximum velocity (TAMV), blood flow volume (BFV), and artery diameter (D). Udder parenchyma images, analyzed by Echovet v2.0, were used to evaluate the mean value (MV), standard deviation (SD), gradient mean value (GMV), gradient variance (GV), contrast (Con), entropy (Ent), gray value distribution (GVD), run length distribution (RunLD), and long run emphasis (LRunEm). In the involution phase, the PI was higher in group B compared to group A (p ≤ 0.05). The PI and RI were higher postpartum, whereas the EDV, TAVM, and D were higher in the transition phase (p ≤ 0.05). Neither the period nor the ewe group affected the MV, SD, GMV, GV, Con, and GVD values (p ≤ 0.05). In the steady state, the LRunEm was higher in group B, but postpartum, it was higher in group A (p ≤ 0.05). In conclusion, B-mode and Doppler can reveal differences (i) between healthy ewes and ewes with subclinical mastitis and (ii) among the different periods studied. Further research is needed on the blood flow and echotexture indices of the udders of ewes with unilateral subclinical mastitis.

Comparison of 5 Versus 7-Day Ovsynch + Progesterone Releasing Intravaginal Device Protocols (PRID) and a Modified G7G with an Option of Heat Detection Protocol for 1st Service in Lactating Dairy Cows.

The aim of this study was to evaluate the efficacy of two timed-AI (TAI) protocols (Group G5D, GnRH and PRID -5d- PGF2a -1d- PGF2a -1d- GnRH, n = 105 and Group G7D, GnRH and PRID-7d- PGF2a -1d- PGF2a -1d- GnRH, n = 98) and a modified G7G protocol combining heat detection (HD) and AI or TAI if HD failed (Group HD, GnRH and PRID -7d- PGF2a -1d- PGF2a -5d- HD or 5d TAI if no HD, n = 92). Pregnancy per AI (P/AI) did not differ between G5D and G7D protocol (G5D: 33.8% vs. G7D: 35.2%, P = 0.85). Cows assigned to G5D and G7D group were pooled as TAI group (GTAI) and further compared to GHD. Within the GHD, more primiparous cows exhibited estrous signs compared to multiparous cows (70.4% vs. 46.2%, P = 0.03). Furthermore, 49 cows (53.3%) were served after HD, whereas 43 cows (46.7%) were served after TAI. There was no difference in P/AI between cows served after HD (51.6%) or after TAI (43.0%, P = 0.49). GHD showed higher P/AI at 1st service compared to GTAI (49.1% vs. 36.4%, P = 0.04), whilst, median days to pregnancy did not differ between the two groups. Overall, P/AI of primiparous cows tended to be better in comparison with multiparous cows (48.3% vs. 37.2%, P = 0.06). In conclusion, there was no significant difference regarding the efficacy of 5- and 7-day Ovsynch + PRID protocols. Moreover, a modified G7G protocol, with intermediate heat detection, resulted in overall better P/AI compared to TAI protocols and appears as a promising strategy to optimize estrus detection for 1st AI.

A comparison study between the results of the Siemens ADVIA 120 analyzer and the manual method for differential leukocyte counts in sheep.

BACKGROUND: The ADVIA 120 is a widely used hematology analyzer, which has not been previously validated for determining differential leukocyte counts in sheep. OBJECTIVES: We aimed to compare differential leukocyte counts on the ADVIA 120 (A-Diff) with counts obtained using the manual method (M-Diff) in sheep. METHODS: Ethylenediaminetetraacetic acid-anticoagulated blood samples analyzed within 4 hours of collection were used. Samples with inappropriately filled tubes, detectable clots, overtly erroneous ADVIA peroxidase cytograms, and poor-quality blood smears were excluded from the study. Two independent observers compared the results of the A-Diff with those of the M-Diff. The M-diff was performed by counting 200 leukocytes on a blood smear. RESULTS: Overall, 88 samples (44 rams and 44 ewes) were included. The correlation between the A-Diff and M-Diff was high for neutrophils (r = .873, P < .001), lymphocytes (r = .863, P < .001), and eosinophils (r = .750, P < .001), and low for monocytes (r = .212, P = .048). The Passing-Bablok regression analyses revealed constant error for eosinophils [1.17%; 95% confidence interval (CI): 0.67%, 1.55%] and proportional error for lymphocytes (0.84; 95% CI: 0.74, 0.95) and eosinophils (0.85; 95% CI: 0.74, 0.96). The Bland-Altman analyses revealed negative biases of 2.4% and 3.0% for neutrophils and lymphocytes, respectively, and positive biases of 3.2% and 0.8% for monocytes and eosinophils, respectively. The extremely low basophil percentages precluded a meaningful method comparison for this leukocyte type. CONCLUSIONS: The ADVIA 120 appears to perform well for determining neutrophil, lymphocyte, and eosinophil percentages in sheep as compared with the manual method. However, blood smear examinations are recommended for the confirmation of abnormal differential leukocyte count results.

Hematologic, biochemical, hormonal, and redox status alterations during reproductive activity in adult rams of two breeds.

BACKGROUND: Reproduction is a crucial, energy-consuming physiologic activity, which can temporarily disturb the homeostatic balance of the organism. Although rams are commonly placed in intense breeding programs on contemporary sheep farms, the clinicopathologic and redox status alterations have not been adequately studied during reproductive activity. OBJECTIVES: The objective was to investigate the clinicopathologic and redox status changes in rams during reproductive activity. METHODS: Thirty-seven (19 Chios and 18 Florina breed) adult, healthy rams were individually mated with ten estrus-synchronized ewes of the same breed. Blood samples were collected prior to mating, during mating, and postmating. CBC (Advia 120), cytologic blood smear evaluation, biochemical analyses (Flexor E, AVL 9180 Electrolyte Analyzer), and thyroxine measurements (Immulite 1000) were performed. The reactive oxygen metabolites (d-ROMs) test, the biological antioxidant potential (BAP) test, and oxidative stress index were used to assess redox status. The linear mixed effects model (statistical language R) was used for statistical analyses. RESULTS: Many, mostly breed-independent, clinicopathologic changes were detected during reproductive activity. Conversely, redox status alterations were found to be breed-dependent. CONCLUSIONS: Intense reproductive activity was related to mild, acute muscle damage in both breeds and mild oxidative stress in Florina rams, whereas Chios rams exhibited oxidative stress resistance. The observed clinicopathologic and redox status alterations were generally consistent with those reported during various forms of physical activity and exercise in animals and humans. Intense reproductive activity does not seem to be physiologically innocuous; however, it appears to be a relatively mild muscle-damaging physical activity.

Effects of testicular hemodynamic and echogenicity changes on ram semen characteristics.

This study investigated correlations among ram semen quality parameters, testicular vascularity and echotexture. Measurements were performed biweekly for a year, before (BE) and after ejaculation (AE), on seven Chios breed rams [three rams (group A) and four rams (group B) of 2-6 and 9-13 years old, respectively], considering breeding (1) and non-breeding period (2). Hemodynamics [Pulsatility Index (PI), Resistive Index (RI)] were evaluated in each testis. Images of testicular parenchyma were analysed to evaluate echogenicity [Mean Value (MV), Contrast (Con), Gray Value Distribution (GVD), Run Length Distribution (RunLD), Long Run Emphasis (LRunEm)]. Semen volume (V), concentration (C), kinetics (CASA), morphology (Sperm Blue), viability (Eosin-Nigrosin) and DNA fragmentation (Acridine Orange) were assessed. In group A/2, semen V was correlated to left testis PI and right testis RI, BE (r = 0.4, p = 0.013/r = 0.6, p < 0.001, respectively), and to right testis PI and RI, AE (r = 0.5, p < 0.001). In A/1/AE, semen C was correlated to left testis PI (r = -0.5, p = 0.003). In group A/1/BE, RI of both testes was correlated to total and tail abnormalities (left: r = -0.5, p = 0.008/ p = 0.004, respectively - right: r = -0.4/r = -0.5, p = 0.01/ p = 0.008, respectively). In group B/1/BE, sperm tail and total abnormalities were correlated to left testis Con (r = 0.5, p = 0.02/ p < 0.001, respectively), whereas in A/2/BE, they were correlated to left testis LRunEm (r = -0.8/r = -0.7, p < 0.001, respectively). In group A/1/BE, DNA damage was correlated to right testis LRunEm (r = -0.6, p < 0.001) and tail abnormalities to left testis PI (r = -0.5, p = 0.03). In group B/1/AE, right testis LRunEm, GVD, RunLD, were correlated to sperm DNA integrity (r = -0.9, p < 0.001). Immotile spermatozoa and viability were correlated to left testis Con in group B/2/AE (r = -0.6/r = 0.6, p < 0.001) and to right testis MV in group A/1/AE (r = -0.6/r = 0.6, p < 0.001). In conclusion, alterations in both ram testes blood flow and echotexture seem to be moderately correlated with sperm assessed parameters, depending on season and ram age.

Combined and breed-specific RIs for hematologic, biochemical, and hormonal analytes in Chios and Florina adult rams.

BACKGROUND: Clinical pathology investigations are essential to evaluate the health status and practice management of ruminants. Appropriate and accurate RIs are required for the interpretation of laboratory results. Despite substantial phenotypic differences and management-related disparities between male and female sheep, RIs in rams are sparse and limited by small reference populations and outdated methods. OBJECTIVES: The purpose of this study was to establish RIs for hematologic, biochemical, and hormonal analytes in adult rams using controlled preanalytic and analytic procedures, and to investigate whether breed-dependent partitioning of RIs is required. METHODS: Reference individuals were selected by a direct a priori method. Blood was collected from 128 healthy adult rams representing 2 Greek breeds (64 Chios and 64 Florina). Hematologic analysis was performed on the Advia 120, while differential WBC counts were manually performed on Giemsa-stained blood smears. Flexor E, AVL 9180 Electrolyte Analyzer, and Immulite 1000 were used for biochemical and hormonal analyses, respectively. Combined and breed-specific RIs were calculated using nonparametric and robust methods, respectively. RESULTS: Five outliers were detected based on laboratory results. Partitioning of RIs, based on both statistical and nonstatistical criteria, was recommended for some of the analytes. CONCLUSIONS: The RIs are in some agreement with those in the literature. The observed differences may be attributed to preanalytic and analytic factors. The RIs determined in this study can be used as a guide for the interpretation of laboratory results in rams and can potentially be adopted by veterinary laboratories, provided that similar equipment and reagents are used.

Evaluation of the in-field efficacy of oregano essential oil administration on the control of neonatal diarrhea syndrome in calves.

The objective of this study was to evaluate under field conditions, whether daily administration of oregano essential oil is effective in preventing and/or diminishing the severity of neonatal diarrhea syndrome in calves aged less than 15days. Ninety-one newborn calves from three dairy farms were assigned into two groups; "Eco" group (n=46) calves were drenched with Greek oregano (Origanum vulgare ssp. Hirtum) essential oil (ECODIAR® liquid 5%) at the dose of 12.5mg/kg body weight once per day until the age of 10days. "Conts" group (n=45) calves were left untreated and served as controls. All animals were monitored daily for the incidence of diarrhea until the age of 15days and their fecal score was recorded. Fecal samples were collected on days 3, 6 and 10 for microbiological and parasitological evaluation. Average fecal score throughout the experiment, incidence of diarrhea, duration and severity of diarrhea episodes were significantly lower in Eco group compared to the controls. Daily administration of oregano essential oil in calves for the first 10days of their life effectively diminishes the severity of naturally acquired diarrhea under field conditions and, under certain hygiene practices, possess a preventive effect against neonatal diarrhea syndrome.

Epidemiological characteristics and clinicopathological features of bluetongue in sheep and cattle, during the 2014 BTV serotype 4 incursion in Greece.

During 2014, an outbreak of Bluetongue virus (BTV) infections attributed to serotype 4 occurred in Greece and spread to south-eastern Europe. In the present article, the clinical and epidemiological data of 15 sheep flocks and 5 dairy cattle herds affected in Greece are described. In sheep, the most frequent clinical signs observed were fever, hyporexia, and edema of the face. A number of clinically affected sheep had chronic laminitis resulting in chronic lameness. Confirmation of suspect clinical cases was performed using BTV-specific real-time RT-PCR, and serotype 4-specific RT-PCR. The average morbidity of bluetongue in the sheep flocks was estimated to be 15.3 % (95 % C.I. 6.8-23.8 %) and the average mortality and case fatality were 4.5 % (95 % C.I. 1.5-7.6 %) and 32.0 % (95 % C.I. 18.1-42.9 %), respectively. The BTV seroprevalence and the ratio of clinical manifestations-to-infections determined in seven of these flocks, were on average 36.5 % (95 % C.I. 15.7-57.3 %) and 24.6 % (95 % C.I. 12.8-36.3 %). BTV ratio of clinical manifestations-to-infections was higher in the imported western European sheep breeds examined compared to the local ones. In dairy cattle, the average herd prevalence of viremia was 48.8 % (95 % C.I. 15.3-82.4 %) and none had signs associated with bluetongue. The results of this study indicate that the 2014 Greek BTV-4 has significant impact on the health status and the viability of sheep in affected flocks but does not cause clinical signs in cattle, despite the high prevalence of viremia.

Evidence of Schmallenberg virus circulation in ruminants in Greece.

During March 2013, we investigated the presence and the levels of Schmallenberg virus (SBV) circulation in three dairy cow herds and three sheep flocks in Central Macedonia, Greece. In two cow herds, a high number of abortions had been observed during the winter. Six bulk-tank milk samples and 147 individual sera were screened for SBV-specific antibodies by ELISA. Positive reactions were obtained from 5 out of 6 bulk-tank milk samples, 58 out of 90 sera from the 3 cow herds, and 2 sera from 2 of the 3 sheep flocks. Twenty-two ELISA-positive sera were tested by serum neutralization test (SNT). SNT confirmed the presence of neutralizing antibodies against SBV in all samples tested, with titers ranging between 1:32 and ≥1:256. No neutralizing antibodies against Akabane virus (AKAV) or Shamonda virus (SHAV) were detected, indicating that neutralizing antibodies against SBV do not cross react with AKAV or SHAV in SNT. ELISA testing of bulk-tank milk samples proved to be convenient and reliable. None of the tested sera was found positive for SBV by real-time RT-PCR, indicating that the sampling was conducted past the viremia stage. This is the first report of SBV circulation in Greece.

Lag effect of microclimatic conditions on DNA integrity of frozen-thawed bovine sperm.

The objective of this study was to investigate the effect of microclimatic conditions on DNA integrity of bovine sperm. DNA fragmentation of frozen-thawed sperm was analyzed in monthly semen samples of nine AI bulls ejaculating in weekly routine for one year. The extent of DNA fragmentation was determined using the Sperm Chromatin Structure Assay (SCSA™) and quantified by the DNA fragmentation index (DFI) and the percentage of sperm showing high DFI values (%DFI) immediately (0h) and 3h (3h) after thawing. Microclimatic factors (ambient air temperature, relative humidity, dew point) were recorded in hourly intervals throughout the year. Their mean values were calculated for seven different time periods (up to day 56) preceding ejaculation (day 0). DFI (P<0.01) and its relative change between 0h and 3h (P<0.05) were related to the set of microclimatic parameters recorded the last 11 days and from day 49 to 43 prior to ejaculation, respectively. A significant relationship was detected between %DFI and microclimatic parameters of days 35-29 preceding ejaculation (P<0.05), while the degree of change of %DFI from 0h to 3h was related to microclimatic parameters recorded from day 28 to 22 before ejaculation (P<0.05). Dew point and relative humidity appeared to be the strongest and weakest predictors of DNA integrity, respectively. In conclusion, the results of the present study showed a lag effect of microclimate on DNA integrity of frozen-thawed bovine sperm. However, microclimatic parameters of a single time period before ejaculation could not be identified as the source of variation of different SCSA parameters.

Gender-related effects of prenatal administration of estrogen and progesterone receptor antagonists on VEGF and surfactant-proteins and on alveolarisation in the developing piglet lung.

BACKGROUND: Vascular endothelial growth factor (VEGF) is essential for embryonic lung development and has been shown to be regulated by estradiol (E2) and progesterone (P). AIM: To investigate the effects of prenatal E2 and P withdrawal by specific receptor antagonists on the mRNA expression of VEGF, surfactant proteins (SP-B and SP-C) and on alveolarisation in lung tissue of male and female pig fetuses. METHODS: Fetuses from 10 sows were randomized to receive either both an intramuscular injection of the E2 receptor blocker ICI 182.780 and the P receptor blocker RTI 3021-022 (ICI+RTI, n=5) or a placebo injection (n=5) at 90 days of gestation (DOG, 115=term). After delivery by cesarean section on 114 DOG, tissue of the left lingula of the piglet's lung (28 placebo, 26 ICI+RTI) was obtained to determine the mRNA expression of VEGF, SP-B and SP-C. Lungs from 15 placebo and 14 ICI+RTI group piglets were removed and alveolar counts performed. RESULTS: The ICI+RTI group showed significantly lower SP-C mRNA expression and alveolar counts compared to the placebo group (p=0.04 and 0.03, respectively). Diminished alveolarisation in the ICI+RTI group was mainly due to the reduction of alveolar counts in male piglets (p=0.02). Within the placebo group VEGF and SP-B mRNA expression in male piglets were significantly lower compared to female piglets (p=0.01 and 0.004, respectively). ICI+RTI treatment abolished this gender-related difference. CONCLUSION: Estradiol and P antagonism affected gender-related differences of key proteins for pulmonary function and development and especially in males was associated with diminished alveolarisation.

Cyclic changes in endometrial echotexture of cows using a computer-assisted program for the analysis of first- and second-order grey level statistics of B-Mode ultrasound images.

The aim of this study was to test the suitability of a computer-assisted echotexture analysis program for analysing first- and second-order grey level statistics of grey levels of B-Mode ultrasound images to examine morphologic changes in the endometrium during oestrous cycle in cows. Four Simmental cows were examined for two consecutive oestrous cycles. Echotexture of the endometrium was assessed by mean grey level (MGL) and homogeneity (HOM) of digitised B-Mode images of the uterine body and both uterine horns. As there were no differences (P>0.05) in MGL and HOM, respectively, between the images of the uterine body and the uterine horns, the mean values of all endometrial images were used for subsequent analyses of MGL and HOM. The factor 'day of oestrous cycle' showed a highly significant (P<0.0001) effect and the factor 'cow' showed a significant (P<0.05) effect on MGL and HOM. No differences (P>0.05) in both echotexture parameters were measured between oestrous cycles within cows. MGL was negatively related to HOM (r=-0.66; P<0.0001). Low MGL and high HOM occurred on Day 0 (=ovulation) and between Days -3 and -1. While MGL was consistently (P>0.05) low between Days -3 and -1, significant changes of HOM with maximum levels on Day -2 (P<0.05) were observed during this time. MGL was consistently (P>0.05) high between Days 4 and 13 while HOM was consistently (P>0.05) low between Days 2 and 13. From Day -3 to Day -1 (r=0.48; P<0.05) plasma oestrogen levels were correlated with HOM, but not with MGL (P<0.05). The results of this study show that changes in endometrial morphology of cows can be measured using a computer-assisted texture analysis program.

Examination of cyclic changes in bovine luteal echotexture using computer-assisted statistical pattern recognition techniques.

B-mode sonography is a well-established diagnostic tool for determination of cycle stage in gynaecology. The aim of this study was to determine whether computer-assisted texture analysis of B- mode sonographic images of bovine luteal glands provides further information about the animal's plasma progesterone concentration and cycle stage. Four Simmenthal cows were examined during two consecutive estrous cycles with an ultrasound device equipped with a 7.5MHz microconvex probe. During each examination three B-mode images of the corpus luteum (CL) were digitized and analyzed off-line using a computer-assisted texture analysis program. Size, echogeneity, and echotexture of the CL were characterized by the following texture parameters: area of cross-sectional planes of the CL (A), mean gray level (MGL), correlation (CORR), run percentage (RPERC), and long-run emphasis (LREM). Plasma progesterone levels (P4) were also determined. All parameters showed characteristic changes during the estrous cycle (P<0.05). Variance component estimates for the effect of Day of estrous cycle on A, MGL, CORR, RPERC, and LREM were 56.6%, 64.6%, 77.6%, 89.9%, and 86.0%, respectively, and 20.6%, 24.5%, 7.2%, 0.0%, and 14.0% for the influence of the individual cow. The factor estrous cycle within cows was responsible for 22.8%, 10.9%, 15.2%, 10.1%, and 0.0% of the variability of A, MGL, CORR, RPERC and LREM values, respectively. Cyclic changes were similar in A and P4. In contrast to P4, which decreased already between Days -5 and -3 (Day 0=ovulation), A stayed at constant high values until Day -3. Mean MGL values were higher (P<0.05) on Days 7, 9, and 13 compared to Days 3 and -3. Mean CORR values were constantly high (P>0.05) during the first days after ovulation and decreased continuously (P<0.05) between Days 5 and 13. Thereafter, mean CORR values remained low (P<0.05) until the next ovulation, except on Day -3 (P<0.05). Mean RPERC rose between Days 1 and 9 from low to high values (P<0.0001) remained at these high values (P>0.05) between Days 9 and 15, and decreased (P<0.05) afterwards to baseline values on Day -1. Mean LREM inclined steeply (P<0.0001) from minimum to maximum between Days 1 and 5. From Days 7 to -3, mean LREM remained (P>0.05) at a constant level close below the maximum value, and decreased to baseline values on Day -1. The results of this study show that statistical pattern recognition techniques provide new information about the luteal glands, thus facilitating a more accurate differentiation between different cycle stages in cows.

Prenatal estrogen and progesterone deprivation impairs alveolar formation and fluid clearance in newborn piglets.

Exposure to high levels of estradiol (E2) and progesterone (P) derived from the fetoplacentomaternal unit during the last trimester of pregnancy may play a crucial role in prenatal lung development and immediate postnatal alveolar fluid clearance (AFC). To measure prenatal alveolar formation and postnatal amiloride-sensitive AFC after pharmacological deprivation of E2 and P in utero, fetuses from five sows received an intramuscular depot injection of the E2 receptor blocker ICI 182.780 (ICI) and the P receptor blocker RTI 3021-022 (RTI) and fetuses of five other sows received a placebo injection (control group) during a laparotomy at 90 d of gestation (term gestation, 115 d). Piglets were delivered by cesarean section on d 114 of gestation. Of 95 live-born piglets, 35 were mechanically ventilated. The airways of the right lower lobe were isolated by a balloon catheter wedged in the bronchus and 5% albumin in 0.9% NaCl with or without 1 mmol/L amiloride was instilled. Amiloride-sensitive AFC was calculated from the protein concentration changes in fluid recovered after 120 min as the percentage of absorbed fluid. Lungs were removed under standardized conditions to perform alveolar counts. Prenatal treatment with ICI and RTI resulted in a significantly lower amiloride-sensitive AFC (median, 31%; min-max, -4-58) than placebo (74%, 18-231). Median alveolar counts per visual field were significantly lower in piglets that were exposed to ICI and RTI (38, 21-78) compared with placebo (56, 32-113). We conclude that prenatal E2 and P deprivation significantly impaired alveolar formation and amiloride-sensitive AFC.

Effects of alpha- and beta-adrenergic receptor stimulation and oxytocin receptor blockade on milking characteristics in dairy cows before and after removal of the teat sphincter.

Alpha (alpha)- and beta (beta)-adrenergic receptors in the bovine mammary gland are mainly present in the teat muscles and in the region where large milk ducts reach the cisternal cavities. The aim of the study was to test the hypothesis that the region of the large mammary ducts is the most important location of alpha- and beta-adrenergic receptor stimulation affecting milk ejection and milk removal. Effects of alpha- and beta-adrenergic receptor stimulation and of oxytocin (OT) receptor blockade on milking characteristics were tested in six cows. Milk flow was measured before and after the distal part of one teat, including the teat canal and teat sphincter, had been partly amputated. Before the operation, milk yield and peak flow rate decreased during alpha-adrenergic receptor stimulation and during the OT receptor blockade, and increased during beta-adrenergic stimulation. After removal of the teat tip, relations of milk yield and peak flow rates after administration of alpha- and beta-agonists and after application of an OT receptor blocking agent were similar to those before operation. Only total milk yield had decreased in the teat-amputated quarter owing to unhindered flow of cisternal milk before cluster attachment. Since responses to alpha- and beta-adrenergic receptor stimulation as well as to OT receptor blockade do not differ with or without the teat sphincter, it is concluded that milk flow is mainly influenced by the muscle tone of the large mammary ducts.