The effect of insulin and relaxin upon mitosis (in vitro) in mammary tissue from pregnant and lactating pigs

The rate of cellular proliferation in the mammary glands of pigs during late gestation and lactation was assessed by measuring the incorporation of ³H-thymidine (T₁) into the DNA of mammary gland explants in vitro. The T₁ showed a linear response over the first 9 hr in vitro, and was not affected by the addition of 500 ng insulin/ml medium. From day 100 to parturition the T₁ rose, reached a peak at 2 d after parturition and declined during lactation to the lowest levels seen at day 21 of lactation.

The inclusion of 0–1000 ng relaxin/ml medium on T₁ at 24–72 hr in vitro had no effect in stimulating T₁ in mammary tissue explants taken from either pregnant or lactating pigs.     

TCDCA对IL-1β刺激下AA大鼠成纤维样滑膜细胞中PGE2分泌的影响

[目的] 进一步探讨牛磺鹅去氧胆酸（taurochenodeoxycholic acid,TCDCA）在抗炎免疫方面的潜在调节作用。[方法] 以AA大鼠成纤维样滑膜细胞作为研究对象,采用ELISA方法检测TCDCA和IL-1β作用下AA大鼠成纤维样滑膜细胞上清液中PGE₂的含量,分析TCDCA对IL-1β刺激下AA大鼠成纤维样滑膜细胞中PGE₂分泌情况的影响。[结果] TCDCA能够对IL-1β刺激下AA大鼠纤维样滑膜细胞PGE₂的分泌产生下调作用（P<0.05）。[结论] TCDCA对IL-1β刺激下AA大鼠成纤维样滑膜细胞中PGE₂的分泌具有抑制作用,为TCDCA在兽医临床应用提供依据。     

Effects of pituitary adenylate cyclase-activating polypeptide (PACAP), prostaglandin E2 (PGE2) and growth hormone releasing factor (GRF) on the release of growth hormone from cultured bovine anterior pituitary cells in vitro

The effect of pituitary adenylate cyclase-activating polypeptide (PACAP) on growth hormone (GH) release was compared with that of prostaglandin E₂ (PGE₂) and growth hormone releasing factor (GRF) from cultured bovine anterior pituitary cells in vitro. Both PACAP and PGE₂ stimulated GH release at concentrations as low as 10⁻⁹ and 10⁻⁸ M, respectively, (P<0.01). However, GRF released GH at a concentration as low as 10⁻¹³ M (P<0.01). Percent increases of GH compared with controls were not significantly different among GRF, PACAP, and PGE₂ at 10⁻⁷ M; however, the increases of GH by the 10⁻⁸ M GRF, PACAP and PGE₂ were 196, 118, and 27%, respectively, (P<0.01), and 124, 65, and 1% in the 10⁻⁹ M media, respectively, (P<0.01). When GRF and somatostatin (SS) were added together, the GH releasing effect of GRF was blunted (P<0.01). Similar bluntness were observed in PACAP and PGE₂, when SS was added. The stimulatory effects of GRF and PGE₂ together were similar to that by either GRF or PGE₂ alone. When GRF and PACAP were added together, the GH released by both secretagogues was greater than that by PACAP alone (P<0.01); however, a synergistic effect was not clear when compared with GRF alone.

These findings suggest that PACAP and PGE₂ may modulate the release of GH in cattle.     

CpG-ODN enhances mammary gland defense during mastitis induced by Escherichia coli infection in goats

Seven healthy native goats in early lactation, weighing 30–40 kg, were used in this study. The right mammary gland of the seven does were infused with CpG-ODN at a dosage of 100 μg kg⁻¹ body weight on the day 5 postpartum (PP). The left glands were used as controls and infused with sterile phosphate-buffered saline (PBS). On day 8 PP, the same dosage of CpG-ODN or PBS was again infused. On day 9 PP, the mammary glands (both right and left) of the seven does were infused with 6 × 10⁶ colony-forming units (CFU) Escherichia coli and, at 0, 8, 16, 24, 48 and 72 h postinfection (PI), milk samples were collected from all glands. Goats were euthanized at 72 h PI and the mammary tissue harvested. Infusion with 6 × 10⁶ CFU ml⁻¹ E. coli induced acute mastitis. Histopathological evaluations showed that polymorphonuclear neutrophils (PMNs) were still present in alveoli at 72 h PI, but PMNs in the CpG-ODN-treated glands has disappeared. Bacteria counts in milk peaked at 16 h PI and CpG-ODN induced a significant decrease in viable bacteria from 16 h PI until the end of the experiment. This study showed that CpG-ODN promoted the expression of its specific receptor (TLR-9 mRNA) in mammary tissue, stimulated IL-6 production, reduced bacteria counts in milk, attenuated the impact of inflammation mediators on cells and significantly shortened the inflammation course. These results suggest that the CpG-ODN improved mammary gland defense and, thereby, had a beneficial effects against mastitis caused by E. coli infection in goats.     

Correlation between carcass conformation and fat cover degree, and muscle fatty acid profile of yearling bulls depending on breed and mh-genotype

The objective was to study the relationships between the actual European beef carcass classification scale, which classifies carcasses with regard to conformation and degree of fat cover scores, and muscle fat quality, depending on breed and mh-genotype. For this purpose samples from 100 yearling bulls from “Asturiana de los Valles” (24 AV(mh/mh), 26 AV(mh/+), 25 AV (+/+)) and “Asturiana de la Montaña” (25 AM) were analysed. The results of the study showed that breed or genotype affect carcass measurement scores and muscle fatty acid profile through its important effect on animal overall fatness. Homozygous double-muscled animals produced carcasses with high conformation and low intramuscular (IM) fat content. While early-maturing and rustic AM animals produced low carcass yield and high IM fat content. The other genotypes (mh/+, +/+) showed, in general, intermediate characteristics. Referring to correlations, carcass conformation was negatively related to saturated (SFA) (r = − 0.69, P < 0.001) and monounsaturated fatty acid (MUFA) (r = − 0.69, P < 0.001) groups, and positively to polyunsaturated (PUFA) (r = 0.72), n-6 (r = 0.72), n-3 (r = 0.71) and unsaturated fatty acid (UFA) (r = 0.69) groups, being all of them significant (P < 0.001). However, carcass degree of fat cover was positively related to SFA (r = 0.53, P < 0.001) and MUFA (r = 0.62, P < 0.001), and negatively to PUFA (r = − 0.61), n-6 (r = − 0.60), n-3 (r = − 0.62) and UFA (r = − 0.53) groups, being all of them significant. Moreover, simple and low-cost prediction equations were calculated for a rapid and sufficiently accurate fatty acid group (SFA, MUFA, PUFA, n-6, n-3, UFA) estimation (R² > 0.46, P < 0.001). In general, meat obtained from double-muscled animals display a more appropriate IM fatty acid profile from the nutritional point of view according to actual recommendations, but it could happen the disability of these lean animals to deposit sufficient IM fat to ensure consumer overall liking or acceptability.     

Pattern of change in the concentration of milk acetone and its association with ovarian activity for pasture-based cows in early lactation

The variation in milk acetone concentration between cows and the resumption of ovarian activity was determined in 68 Holstein–Friesian cows grazing pasture. Cows calved in two groups approximately three weeks apart and were assessed over the first 60 d of lactation.

The largest coefficient of variation (152%) for milk acetone concentration between cows was found in the first week of lactation. This variability in milk acetone concentration was largely explained by a consistent cyclic pattern, in relation to the day of the study, with regular peaks at 21–26 d intervals. These peaks were synchronised between cows, and cows in the second group synchronised with cows in the first group.

Milk acetone peaks greater than 0.4 mmol/L were associated with a significant drop in milk yield, possibly due to energy deprivation around this time. In this respect, liveweight change over the first 60 d of lactation was significantly negatively correlated (r = − 0.29, p < 0.05) with the mean milk acetone concentration taken over the same period. However, there was no significant relationship (p > 0.05) between mean or maximal milk acetone concentration and the resumption of ovarian activity for the cows that resumed ovarian activity in the first 9-weeks post-partum.

The results of this study suggest that the use of milk acetone as a practical indicator of energy balance is limited due to its variability over time. Milk acetone concentration may be useful in experimental situations or when regular milk samples can be obtained in the field, such as in-line analysis. There is a need to undertake more research to elucidate the implications of the cyclic pattern of acetone concentration in milk on reproductive performance.     

The use of even-chain alkanes sprayed onto herbage as rate of passage markers in goats

An experiment was conducted with Saanen goats fed fresh grass ad libitum to compare ⁵¹Cr-mordanted fibre and even-chain alkanes sprayed onto either grass leaves or stems to estimate faecal output, total mean retention time (TMRT) and parameters obtained from faecal marker concentration curves, such as k₁ (slow rate of passage), k₂ (fast rate of passage) and transit time (TT). ⁵¹Cr-mordanted grass showed the lowest fractional rates of passage (k₁ and k₂) and hence the largest value of TMRT. There were not significant (P > 0.05) differences between even-chain alkanes sprayed onto leaves or stems for k₂, TT and TMRT, but k₁ estimates were higher (P < 0.05) for stems than for sprayed leaves. Despite the marker used, TMRT values were negatively correlated with the level of dry matter intake (r = − 0.81, − 0.80 and − 0.80 for ⁵¹Cr-mordanted fibre and even-chain alkanes adsorbed onto leaves or stems, respectively). Average faecal outputs estimated from faecal concentrations of ⁵¹Cr-mordanted fibre and even-chain alkanes were not different from the actual outputs but there were differences between markers in the accuracy of estimation. The highest mean square prediction error (MSPE) and the poorest correlation between observed and estimated faecal output values corresponded to even-chain alkanes adsorbed onto stems. Values estimated using ⁵¹Cr-mordanted fibre and even-chain alkanes adsorbed onto leaves were significantly correlated with faecal outputs (r = 0.94 and r = 0.92, respectively), with MSPE being greater for the latter marker.     

Dose effect of human growth hormone-releasing factor and thyrotropin-releasing factor on hormone concentrations in lactating dairy cows

Two experiments were conducted to study the effects of growth hormone-releasing factor (GRF) and thyrotropin-releasing factor (TRF) administration on hormone concentrations in dairy cows. In the first trial, 12 cows were used on 5 consecutive days to determine the effect of four sc doses of GRF (0, 1.1, 3.3 and 10 μg•kg⁻¹ BW) and three sc doses of TRF (0, 1.1 and 3.3 μg•kg⁻¹ BW) combined in a factorial arrangement. GRF and TRF acted in synergy (P = .02) on serum growth hormone (GH) concentration even at the lowest dose tested and GH response to the two releasing factors was higher than the maximal response observed with each factor alone. TRF increased (P<.01) prolactin (Prl), thyrotropin (TSH), triiodothyronine (T₃) and thyroxine (T₄) concentrations similarly at the 1.1 and 3.3 μg•kg⁻¹ doses and GRF did not interact (P>.40) with TRF on the release of these hormones. In the second trial, the effect of GRF (3.3 μg•kg⁻¹ BW, sc) and TRF (1.1 μg•kg⁻¹ BW, sc) was tested at three stages (18, 72 and 210 days) of lactation on serum Prl and TSH concentrations. Eighteen cows (n = 6 per stage of lactation) were used in two replicates of a 3 × 3 latin square. The TRF and GRF-TRF treatments were equipotent (P>.05) in increasing Prl and TSH concentrations. Prl and TSH responses were similar (P>.40) throughout lactation. In summary, GRF at doses ranging from 1.1 to 10.0 μg•kg⁻¹ and TRF at doses ranging from 1.1 to 3.3 μg•kg⁻¹ act in synergy on GH release and do not interact on Prl, TSH, T₃ and T₄ concentrations in dairy cows. Furthermore, Prl and TSH response to TRF are not affected by stage of lactation.     

Effect of exogenous thyrotropin-releasing hormone (TRH) administration on plasma levels of triiodothyronine (T3), thyroxine (T4) and growth hormone (GH) in chronically catheterized suckling piglets.

The purpose of the present study was to determine experimental conditions to stimulate secretion of thyroid hormones (T₃ and T₄) with thyrotropin-releasing hormone (TRH) injections in suckling piglets during the first weeks of postnatal life. Three consecutive experiments were conducted. Four 10–20 d old piglets were i.m. injected with 0, 20, 100, 500 μg (experiment 1) or 0, 4, 20, 100 μg TRH/kg BW (experiment 2) according to a 4 × 4 latin square design involving different litters in each experiment. Blood samples were taken −15, −1, 15, 30, 45, 60, 90, 120 180 and 300 min after TRH injection in experiment 1, and −.25, −.08, .25, .5, 1, 2, 4, 6, 8, 12, 24, 30, 36, 48, 60 and 72 hr after TRH injection in experiment 2. T₃ and T₄ levels were significantly (P<.01) increased as soon as 30 and 45 min after TRH injection, respectively. Maximal levels of T₃ and T₄ were obtained 2 and 4 hr after the injection of 100 μg TRH. T₃ and T₄ returned to basal levels within 6 and 8 hr post injection, respectively. Plasma pGH levels were significantly (P<.001) increased 15 min after TRH injection in piglets injected with 500 μg. In experiment 3, 100 μg TRH/kg BW were injected i.m. either daily or every other day from .0 to 23 days of age. Results showed that T₄ response to TRH did not decrease after repeated injections. These results indicate that daily i.m. injections of 100 μg TRH/kg BW can be used to increase thyroid hormone levels for at least 13 d in the young suckling piglet.     

Skatole metabolism in the intact pre-pubescent male pig: The relationship between hepatic enzyme activity and skatole concentrations in plasma and fat

The objective of this study was to evaluate, in the pre-pubescent intact male pig, the relationship between skatole levels and the activity of hepatic cytochrome P4502E1 (CYP2E1), cytochrome P4502A (CYP2A), aldehyde oxidase (AO), and phenol sulfotransferase 1A1 (SULT1A1). The activity of these enzymes has been positively associated with skatole clearance in mature boars. Twenty-four intact male pigs were weaned at 28 days of age and slaughtered 2 weeks postweaning, at which time caecal contents, blood, fat, and liver samples were collected. Caecal contents and fat were analyzed for skatole concentrations, and plasma was analyzed for skatole and steroid hormone (testosterone (T), dehydroepiandrosterone (DHEAS), estrone sulphate (E₁S)) concentrations. CYP2A, CYP2E1, and AO, as well as SULT1A1 activities were evaluated in liver samples. Stepwise regression was utilized considering plasma or fat skatole concentration as the dependent variables and hormone concentrations and enzyme activities as independent variables. The activities of the enzymes CYP2A, CYP2E1, and AO and concentrations of the hormones T, DHEAS, or E₁S were not correlated with concentrations of skatole in plasma or fat. However, SULT1A1 activity was negatively correlated with plasma (r = − 0.70, P < 0.05) and backfat (r = − 0.41, P < 0.05) skatole concentrations. Furthermore, this correlation was improved in plasma (r = − 0.88, P < 0.05) and fat (r = − 0.63, P < 0.05) when the concentrations of skatole in caecal contents was included as an independent variable in the multiple regression analysis, demonstrating the importance of measuring skatole production in these studies. T, DHEAS, and E₁S concentrations in plasma were not correlated with the activity of any of the enzymes evaluated. This study suggests that SULT1A1 is important in the metabolism of skatole in pre-pubescent pigs and the overall metabolism of skatole in the pre-pubescent pig differs from that in the mature boar.     

Effects of Saccharomyces cerevisiae on in vivo organic matter digestibility and milk yield in buffalo cows

The influence of a dietary supplementation with Saccharomyces cerevisiae (SC) during the first and the second phase of lactation on dry matter (DM) intake, organic matter digestibility, milk yield and quality and haematological profile was evaluated in buffalo cows. Lactating buffaloes (n = 190), 118.7 days in milk (DIM), were randomly divided into Group C (control, n = 95) and Group T (fed diet supplemented with 98 billion CFU of S. cerevisiae, n = 95). Eight buffaloes for each group (Groups T1 and C1), 85.4 DIM, were used to study the in vivo digestibility and the haematological profile. No differences were found for DM intake (16.5 kg·day^− 1) and haematological profile. The SC supplementation increased milk yield (7.9 ± 0.2 vs. 7.4 ± 0.2; P < 0.01) but did not affect milk fat and protein. SC supplementation increased OM digestibility, mainly, in the first phase of lactation (< 135 days), thus allowing a higher energy availability for milk yield and reduced fat mobilization.     

Differentially expressed genes associated with Staphylococcus aureus mastitis of Canadian Holstein cows

To study pathway specific gene expression within the immune-endocrine axis of dairy cows with Staphylococcus aureus mastitis, mRNA was collected from blood mononuclear cells (BMCs) and milk somatic cells (MSCs) of cows (n = 7) identified as culture positive for S. aureus and their matched negative control cows (n = 7) with no evidence of S. aureus mastitis. Labeled cDNA probes derived from BMCs and MSCs of infected and healthy cows were applied to a bovine immune-endocrine cDNA array containing 167 genes. Genes with a log₂ ratio ≥ 0.5 were considered to be up-regulated and genes with a log₂ ratio ≤ −0.5 to be down-regulated. In total, 22 genes were differentially displayed in BMCs and 16 genes in MSCs of case versus controls. Expression of selected genes in BMCs and MSCs were confirmed by real-time PCR. The RT-PCR results were highly correlated with microarray measurements. Some of these genes, such as interleukin (IL)-8 have been previously implicated in other bacterial diseases, and are known to regulate immune responses; whereas, others may reflect novel pathways or genes involved in progressive mammary gland disease. For example, IL-18 was up-regulated in BMCs but not MSCs of mastitic quarters, while IL-17 was more highly expressed in MSCs compared to BMCs. This study identified a number of differentially expressed genes associated with bovine S. aureus mastitis and demonstrates the intricacy of the patterns of gene expression that influence host response to a complex pathogen of significant relevance to both human and veterinary medicine.     

烟碱对LPS诱导的兔子宫内膜上皮细胞炎症因子及前列腺素分泌的影响

试验旨在探索α7烟碱乙酰胆碱受体（α7 nicotinic acetylcholine receptor,α7nAChR）的高亲和力激动剂烟碱对脂多糖（lipopolysaccharides,LPS）诱导的兔子宫内膜上皮炎症的作用机制。分离发情后期兔的子宫内膜上皮细胞,用100 ng/mL LPS对细胞进行炎性刺激12 h。用CCK8法检测不同浓度烟碱（5、10和20 μg/mL）对细胞存活率的影响,筛选合适浓度的烟碱进行后续试验。将细胞分为对照组（CON）、LPS、LPS+烟碱、LPS+烟碱+甲基牛扁碱（MLA）（α7nAChR的特异性颉颃剂）组,通过ELISA法检测细胞培养上清液中白细胞介素-1β（interleukin 1β,IL-1β）、IL-6、IL-8、肿瘤坏死因子-α（tumor necrosis factor,TNF-α）、前列腺素E₂（prostaglandin E₂,PGE₂）和前列腺素F_2α（PGF_2α）的含量。试验成功分离兔子宫内膜上皮细胞,且传至第5代仍保持良好的生长状态。CCK8检测结果显示,20 μg/mL烟碱组细胞存活率显著降低（P<0.05）,10 μg/mL烟碱对细胞存活率无显著影响（P>0.05）,所以选择10 μg/mL烟碱进行后续试验。ELISA结果显示,与对照组相比,LPS组IL-1β、IL-6、IL-8、TNF-α、PGE₂和PGF_2α的含量显著增加（P<0.05）;与LPS组相比,LPS+烟碱组显著降低IL-1β、IL-6、IL-8、TNF-α、PGE₂和PGF_2α的含量（P<0.05）,LPS+烟碱+MLA组炎性因子和前列腺素的含量差异不显著（P<0.05）。以上结果表明,烟碱对LPS诱导的兔子宫内膜上皮细胞分泌IL-1β、IL-6、IL-8、TNF-α、PGE₂和PGF_2α具有下调作用,推测烟碱通过α7nAChR介导炎性因子和前列腺素分泌下调而发挥抗炎作用,该结果可为研究α7nAChR作为子宫内膜炎治疗靶点的药物选择提供参考。     

A model to determine sampling strategies and milk inoculum volume for detection of intramammary Staphylococcus aureus infections in dairy cattle by bacteriological culture

A model was developed to evaluate the effects that methods of obtaining milk samples and culture inoculum volumes had on the sensitivity of microbiological culture to detect Staphylococcus aureus intramammary infections (IMI). An assumption was made that milk from mammary quarters infected with S. aureus only contains bacteria intermittently. A modified sine wave function was used to model this intermittent shedding pattern. Specifications for the components of the shedding cycle used in this function were based on quantitative culture results from 54 experimentally infected S. aureus quarters, sampled daily for a period of 30–49 days. The components of the shedding cycle were length in days, peak number of CFU shed per milliliter of milk, and length of time in the cycle when no shedding occurred. These components were used to estimate the model's predicted distribution of S. aureus CFU ml⁻¹ milk when individual quarter milk samples were cultured for S. aureus. The sensitivity of culture for several sampling methods was then calculated. The model predicted that culture of a single quarter milk sample had a sensitivity ranging from 60 to 87% for detection of S. aureus IMI depending on inoculum volume. Quarter milk samples taken on day 1 and repeated either on day 3 or day 4, and cultured separately using 0.1 ml of milk for culture inoculum, were predicted to have sensitivities of 90–95% and 94–99%, respectively. Other milk-sampling strategies examined included culture of a composite milk sample (equal-volume mixture of milk from four separate mammary quarters ) and pooled milk samples in which samples from different milkings (either quarter or composite samples) were mixed together and then cultured. The range of predicted sensitivities of these other sampling strategies was 30–97%. Factors having the greatest impact on the sensitivity of culture, in order of importance were: the type of milk sample, the volume of milk cultured, and the time interval between repeated milk sample collection strategies.     

Temporal and spatial patterns of African swine fever in Sardinia

Temporal patterns and spatial distribution of African swine fever (ASF) were studied through the analysis of routinely collected data in the ASF-endemic area of the Province of Nuoro, Sardinia. During 1993–1996, ASF outbreaks were reported from 45 out of the 82 municipalities of the study area. Overall farm-level incidence rate (IR) was 1.3 outbreaks per 100 farms-year. ASF peaked in 1995 (IR=1.8) and declined in 1996 (IR=0.82). Significant (P<0.05) spring peaks of ASF outbreaks and affected municipalities were detected using statistical methods for circular distributions. Spatial clustering of ASF-affected municipalities, as evaluated by join-count statistics, was significant in 1993 (Z_jc=−3.0, P<0.01) and 1994 (Z_jc=−3.2, P<0.01) but not in 1995 (Z_jc=−0.6, P=0.55) and 1996 (Z_jc=−1.2, P=0.23). Extensive pig farming and ASF were spatially co-distributed (κ=0.51, 95% CI=0.33–0.70).     

Risk factors for intramammary infections and relationship with somatic-cell counts in Italian dairy goats

Routine examination of milk was performed on five herds of lactating goats in northern Italy as part of a milk quality-monitoring program in the year 2000. As part of the study, aseptic samples of foremilk were collected monthly from both half udders during the entire lactation for 305 goats, resulting in a total of 4571 samples. The samples were tested with cytological and bacteriological analyses to evaluate the relationship between mammary infections and somatic-cell count (SCC; Fossomatic (TM) method). Prevalence of intramammary infection (IMI) was 40.2% (n = 1837) of all udder-half samples examined. The most-prevalent mastitis agents were coagulase-negative Staphylococci (CNS), 80% (n = 1474 udder-half samples); within this group, Staphylococcus epidermidis was the most-prevalent species (38%). Other prevalence were Staphylococcus aureus 6% (n = 112 udder-half samples) and environmental pathogens 14% of infected udder-half samples (n = 251) with a diverse mixture of species, none of which had a frequency of >4%. Enterococcus faecalis was the most-frequently isolated among this group. Neither Salmonella spp. nor Listeria monocytogenes were detected. The risk (sample level) of infection differed across herds, parities, and stage of lactation according to results from logistic multiple regression. Infection was more common among goats in third and fourth parities and during the later stages of lactation. Of the 2734 samples from uninfected udder halves, the mean log₂ SCC was 3.9 cell/ml; of the 1837 bacteriological positive samples, the mean log₂ SCC was 5.6 cell/ml. According to results from a linear mixed model, concentrations of somatic cells tended to increase with increasing age and days in milk and with the presence of bacteria. Infection with S. aureus was associated with the highest SCS.     

Effect of a nonsteroidal aromatase inhibitor on in vitro and in vivo secretion of estradiol and on the estrous cycle in ewes.

Three experiments were performed to study effects of decreased concentrations of estradiol-17β (E₂) on lifespan and function of ensuing ovine corpora lutea (CL). In experiment 1, 52 follicles were collected from 10 ewes and placed into individual culture with 0 or .01 μCi ³H-androstenedione (10 ng; ³H-A) and 0, 10⁻¹¹, 10⁻⁹, 10⁻⁷, or 10⁻⁵ M of a nonsteroidal aromatase inhibitor, CGS16949A (CGS). Concentrations of E₂ secreted into the medium, and synthesis of estrogens as estimated by formation of ³H-water from ³H-A were decreased by 10⁻⁵ and 10⁻⁷ (P<.01), but not 10⁻⁹ or 10⁻¹¹ M CGS. In experiment 2, luteolysis was induced in 24 ewes by injection of PGF₂ on days 5 to 10 of the estrous cycle (0 hr). Ewes received 0, 0.5, 1.0, 2.0 or 4.0 mg CGS per kg BW i.v. at −12, 0, 12 and 24 hr, and an ovulatory dose of hCG at 36 hr. Jugular (P<.001) and vena caval (P<.001) concentrations of E₂ were decreased by CGS at all doses tested for 8 to 10 hr, but had returned to levels similar to control ewes by the time of the next injection. Concentrations of E₂ around the time of the LH surge were similar in control and treated ewes. During the subsequent luteal phase, concentrations of progesterone (P₄) were similar in control and treated ewes. Thus, transient decreases in E₂ during the follicular phase were not deleterious to the subsequent luteal phase. In experiment 3, luteolysis was induced in 18 ewes by injection of PGF₂ on days 6 or 7 (0 hr) of the estrous cycle. Ewes received 0 or 1 mg CGS per kg BW i.v. every 8 hr from 0 to 40 hr. Ovulation was induced with hCG at 36 hr. CGS reduced jugular (P<.001) and vena caval (P<.001) concentrations of E₂, prevented an endogenous surge of LH (P<.05) and increased (P<.001) concentrations of FSH. All ewes had ovulated a marked follicle by 72 hr, but onset of the luteal phase, as assessed by concentrations of P₄, was delayed (P<.01) in ewes receiving CGS. Delayed luteal phases were not solely attributable to the presence of new CL or to luteinization of follicular cysts. When data were aligned according to the day ewes were observed in estrus, profiles of P₄ did not differ with treatment. Therefore, normal luteal function ensued following estrus whether or not ewes re-ovulated. In conclusion, decreased secretion of E₂ by the preovulatory follicle was not involved in the ontogeny of CL of short lifespan or subnormal function. Instead, adequate production of E₂ or precisely timed E₂ secretion may be required during follicular development for subsequent functional luteinization.     

前列腺素E2和F2α对LPS刺激后奶牛子宫内膜上皮细胞COX-1和COX-2表达的影响

试验旨在阐明前列腺素E₂（prostaglandin E₂,PGE₂）和F_2α（prostaglandin F_2α,PGF_2α）对体外培养的奶牛子宫内膜上皮细胞中环氧合酶-1（cyclooxygenase-1,COX-1））与环氧合酶-2（cyclooxygenase-2,COX-2）表达的影响。培养奶牛子宫内膜上皮原代细胞和传代细胞,第4代细胞以1×10⁶个/孔接种于6孔板,以10^-7mol/L PGE₂和PGF_2α分别预处理细胞24 h,以100 ng/mL细菌脂多糖（lipopolysaccharides,LPS）刺激细胞4、8和12 h后分别提取RNA和总蛋白质,采用实时荧光定量PCR与Western blotting等技术检测COX-1与COX-2 mRNA和蛋白质的表达量。结果表明,与对照组相比,COX-1 mRNA表达量在PGE₂单独作用4、8和12 h后显著上调（P<0.05）;COX-2 mRNA表达量在PGE₂单独作用4和12 h后显著上调（P<0.05）,PGE₂单独处理使COX-1、COX-2蛋白表达量均显著上调（P<0.05）。与对照组相比,LPS刺激8和12 h时COX-1 mRNA表达量显著下调（P<0.05）,LPS刺激后COX-1蛋白表达量无显著变化（P>0.05）;LPS刺激后4、8和12 h时COX-2 mRNA表达量显著上调（P<0.05）,LPS刺激后COX-2蛋白表达量显著上调（P<0.05）。与LPS单独处理组相比,LPS+PGE₂处理组在8和12 h时COX-1和COX-2 mRNA表达量均显著上调（P<0.05）,同时COX-1和COX-2蛋白表达量也显著上调（P<0.05）。PGF_2α在LPS未刺激和刺激后对COX-1和COX-2 mRNA的表达无显著影响（P>0.05）,仅在PGF_2α单独处理8和12 h后COX-1 mRNA表达量上调（P<0.05）。两种激素联合处理与各自单独处理及LPS单独刺激相比,对COX-1和COX-2 mRNA表达具有一定的协同诱导作用。     

Characteristics of Rabbit Transgenic Mammary Gland Expressing Recombinant Human Factor VIII

The objective of this research was to compare (i) the content of milk protein and recombinant human factor VIII (rhFVIII) in the milk of transgenic and non-transgenic rabbit females at three lactations and (ii) histological structure, ultrastructural morphology and occurrence of apoptosis in rabbit transgenic and non-transgenic mammary gland during third lactation and involution. Significant differences (t_0.05) in milk protein content were found between transgenic and non-transgenic at all three lactations. The percentage of apoptotic cells was significantly higher (t_0.01) in non-transgenic ones compared with transgenic mammary gland tissues (6.5% versus 2.4%) taken at the involution stage. Morphometrical analysis of histological preparations at the involution stage detected a significantly higher (t_0.05) relative volume of lumen in transgenic animals compared with non-transgenic ones (60.00 versus 46.51%). Ultrastructural morphology of the transgenic mammary gland epithelium at the involution stage revealed an increased relative volume of protein globules (t_0.05); at the lactation stage, a significantly higher volume of mitochondria (13.8%) compared with the non-transgenic (9.8%) ones was observed. These results, although revealing differences in some parameters of ultrastructure and histology, indicate no harmful effect of the mouse whey acid protein-hFVIII transgene expression on the state of mammary gland of transgenic rabbit females.     

Impact of milking characteristics and morphology of udder and teats on clinical mastitis in first- and second-lactation Norwegian cattle

Data from the Norwegian progeny testing program were used to examine the impact of milking characteristics and morphology of udder and teats on clinical mastitis in first- and second-lactation Norwegian cattle. The study was designed as a 1-1 matched case-control study with herd, parity, days in lactation and calving season as matching variables. Conditional maximum likelihood logistic regression was used to evaluate the effects of three primary (2 min milk, milk leakage and teat-end-to-floor distance) and six other study variables. Treatment records from the Norwegian health card system on acute and chronic clinical mastitis were used to define cases. The chosen model included 565 matched pairs. Significant risk factors of clinical mastitis were decreasing teat-end-to-floor distance (P = 0.02) and periparturient udder edema (P < 0.01).

Borderline effects were demonstrated by larger than herd-average teat diameter (P = 0.04), udder asymmetry (P = 0.05) and increasing 2 min milk (P = 0.08). Results were compared to a previous study on the same data with log_e somatic cell count as the dependent variable. Inclusion of teat-end-to-floor distance in the genetic improvement scheme may increase the efficiency of genetic selection for mastitis resistance.