Expression of vascular endothelial growth factor receptors and their ligands in rat uterus during the postpartum involution period

Abstract

Vascular endothelial growth factor (VEGF) and its specific receptors, FLt1/fms, Flk1/KDR and FLt4, play important roles in vasculogenesis, and physiological and pathological angiogenesis. Whether angiogenic growth factors are involved in regulating angiogenic processes during the postpartum involution period (PP) of the rat uterus is unknown. We used immunohistochemistry to analyze the expression levels of VEGF, the fms-like tyrosine kinase 1 (FLt1/fms), the kinase insert domain-containing region 1 (Flk1/KDR), Fms-related tyrosine kinase 4 (FLt4) and vascular endothelial growth inhibitor (VEGI) in the rat uterus during the days 1, 3, 5, 10 and 15 of the PP to determine the temporal and spatial expressions of VEGF and its receptors during the PP. Throughout the PP, cytoplasmic and membrane staining of VEGI, VEGF and their receptors were observed in the lumens, crypts and glandular epithelial cells as well as in connective tissue and vascular endothelial and smooth muscle cells in the endometrium. We found that the intensity of the immunoreactions in the endometrium varied with the morphological changes that occurred during involution. Immunoreactions for VEGI, VEGF and their receptor, Flk1/KDR, in the luminal epithelial cells were stronger than those in the glandular epithelial and stromal cells, particularly during PP 1, 3 and 5, which suggests that these peptides may contribute to re-epithelialization of the endometrium. On the other hand, Flt1/fms immunoreactivity was strong mainly in the stromal cells during the PP. The presence of VEGF and its receptors (FLt1/fms, Flk1/KDR, FLt4) in the stromal cells and blood vessels during the PP suggests that they may contribute to regulating stromal repair and angiogenesis in the involuting uterus of the rat.     

Resumption of ovarian follicular activity and uterine involution in the postpartum llama

Resumption of ovarian follicle activity and uterine involution was studied in the post partum llama. Thirty-nine adult multiparous llamas were monitored by ultrasonography and analysis of urinary estrone sulfate for 30 d post partum at the La Raya Research Station in Peru. Uterine involution was measured in terms of reduction of length and diameter of both uterine horns. Correlation analysis was used to relate follicle size and concentration of estrone sulfate. Analysis of variance was used to determine rate of uterine involution relative to days post partum. The left ovary was palpated and scanned by Day 3 post partum in contrast to Day 1 post partum for the right ovary. Ovulatory size follicles, 7 mm, were present by Day 7.4 post partum (range 4 to 14 d). Follicle growth was detected as early as Day 4 post partum with follicle size being less during the first follicle wave (7.4 mm) compared to the second and third waves (9 to 10 mm). Concentrations of urinary estrone sulfate were positively related (P<0.05) to follicular size, but to a lesser degree during the first follicle wave (19.4 ng/mg Cr), than to the second wave (25.4 ng/mg Cr). Uterine involution, as measured by diameter, was different between the left (gravid) and right (nongravid) uterine horn (P<0.05) for the 17 d post partum, and was also different from that of control females for the 21 d post partum. Uterine involution was complete in 63% of females by Day 21 post partum.     

Effect of subclinical uterine infection on cervical and uterine involution, estrous activity and fertility in postpartum buffaloes

Nili-Ravi buffaloes (n=29) that calved normally between August and November and did not develop any clinical reproductive disorder after calving were studied for the incidence of sub-clinical bacterial infection of the uterus and its effects on postpartum reproductive efficiency. The incidence of subclinical uterine infection was 24% (7/29). Involution of the cervix and uterus was slower (P < 0.01) in the infected group than in the normal group (45.6 vs 31.1 days and 46.3 vs 35.8 days), respectively. The mean diameters of cervix and gravid horn on Day 12 post partum and on completion of involution did not differ between buffaloes of the two groups. However, the rate of involution of the cervix and the gravid horn was lower in buffaloes of the infected group (2.2 vs. 2.7 mm/day and 2.6 vs. 3.2 mm/day). The mean interval to first post partum ovulation was similar in buffaloes in the infected (35.5 days) and the normal group (33.8 days). The life span of corpus luteum formed after first ovulation was shorter (11 days) in buffaloes of both groups than that of a normal estrous cycle (15 to 17 days). The incidence of silent ovulation was apparently higher in buffaloes of the infected group (83 vs. 60%) but the difference was not significant. For the first four months after calving, the mean interval to first postpartum estrus was longer in buffaloes of the infected group (73.0 vs. 47.7 days; P < 0.01). Similarly, the average service period was longer in buffaloes of the infected group (91.0 vs. 64.8 days; P < 0.05). The overall pregnancy rate for the first four months after calving did not differ between buffaloes of the two groups. We conclude that subclinical bacterial infection of the postpartum uterus delays the cervical and uterine involution which can, in turn, delay the occurrence of first postpartum estrus and prolong the service period in buffaloes.     

Postpartum ovarian activity,uterine involution and fertility in indigenous buffaloes at a selected village location in Sri Lanka

Reproductive events were monitored over a period of 30 months in a sample of 55 indigenous (Lanka) buffalo cows belonging to five farmers selected at random in a specific village in the dry zone low-country region of Sri Lanka. The animals were maintained under the existing traditional methods of extensive grazing management and low-intensity utilization for milk and draught power in a crop-livestock system. Annual calving rate was 87.2%, with 82.4% of all calvings occurring between December and March (peak calving season). Involution of the uterus, based on rectal palpation, was completed at an interval of (mean ± SD) 32.9 ± 8.2 days post-partum. The majority (70.8%) of first services postpartum were recorded between February and April. The interval from calving to first service was 57.3±49.2 days for 80 animals which calved during the peak calving season and 156.2 ± 95.6 days for eight animals calving outside the peak season (overall 66.4 ± 61.4 days, n = 88). The intervals to uterine involution and first service were correlated (r=0.53, P < 0.01) and both were significantly influenced by month of calving, but not by age or parity of the cow. The interval from calving to palpation of the first corpus luteum was 52.3 ± 16.8 days, and to the first elevation of progesterone above 0.5 ng/ml of fat-free milk was 54.9 ± 17.9 days.Of the 36 cases where complete progesterone profiles and clinical data were available, first services were recorded in 27 (75%) before palpation of a corpus luteum or elevation of progesterone after calving, and conception to this service was confirmed in 24 (88.9%). Elevation of progesterone preceded first service in seven (19%); the resultant luteal phases were short in four and normal in three. Overall conception rate to first service was 77.7% and the number of services per conception was 1.29. The calving to conception intervals ranged from 23 to 502 days (mean 71.5 ± 66.2, mode 41–60, n = 82) and the calving intervals from 329 to 816 days (mean 384.9 ± 62.9, mode 351–370, n=79).These findings confirm that Lanka buffaloes are capable of maintaining high fertility under certain traditional systems of management. The major cause of long calving intervals was prolonged postpartum acyclicity, which occurred mainly in animals calving outside the peak calving season.     

Pattern of arterial supply to the ovaries in Nili-Ravi buffaloes: Relationship with the distribution of ovarian structures

Reproductive tracts of 12 heifers and 18 adult buffaloes were collected from a slaughter house to study gross ovarian morphology and the distribution of follicles and corpora lutea on the surface of the ovary relative to the pattern of the arterial supply. Mean weight and volume of the ovaries were 3.8 ± 0.3 g and 4.08 ± 0.07 cm³, respectively, and were higher in adult buffaloes, but the difference was a function of body weight. Seventy percent of the ovaries were supplied by a single artery while the remaining 30% received two branches of the ovarian artery. Regardless of the position of entry of a single branch, ovarian follicles were located more frequently in the central (41.1%) than in medial (30.7%) or lateral portions of the ovary (28.2%). In ovaries supplied by two branches of the ovarian artery, follicles were distributed more uniformly on the ovarian surface. Number of follicles on the ovarian surface tended to increase when one of these branches was medial in position. Corpora lutea were observed more frequently in the part(s) of the ovary adjacent to the point of arterial entry regardless of the number of branches. Presence of a corpus luteum on the ovary had a negative effect on follicular development. The data suggested a possible relationship of the ovarian activity of the buffaloes with the pattern of arterial supply to the ovaries. Pattern of arterial supply might be contributing to the growth of the follicles, possibly by either saving them from atresia or by stimulating their growth.     

Effect of biostimulation on uterine involution, early ovarian activity and first postpartum estrous cycle in beef cows

The objective was to determine the effect of biostimulation (bull-exposure) on uterine involution (UI), plasma progesterone concentration (P4), size of largest follicle (LF), number of follicles larger than 5 mm ( F > or = 5 ), presence of fluid in uterine lumen (PF), presence of luteal tissue (LT), and length of the first estrous cycle postpartum (LEC). Ninety Angus cows with calves were allocated by parity and body frame into three groups (30 per group) 1 week postpartum. Two groups were exposed to bulls (BE) and one non-exposed group (NE) served as a control. Data were collected during weekly sessions of palpation per rectum, ultrasonography and bleeding on a subgroup of 30 cows (10 cows per group) for 6 weeks, and permanent surveillance of estrus with HeatWatch on all 90 cows. There were no significant differences between BE and NE cows for UI ( 17.1+/-1.1 days versus 20.1+/-1.6 days), LF ( 9.5+/-1.7 mm versus 11.0+/-2.4 mm), F > or = 5 ( 1.20+/-0.3 versus 1.47+/-0.09 ), and PF. However, LT was detected in more BE than NE cows (13 versus 2; P<0.001 ). Overall differences in P4 were found between BE and NE cows with detected LT ( 2.00+/-0.3 ng/ml versus 1.05+/-0.4 ng/ml, respectively; P<0.05 ). More BE cows resumed reproductive cyclicity with estrous cycles normal in length compared with NE cows (16/30, 53%; 16/30, 53%; and 8/30, 26.6%, for the two BE groups and the NE group, respectively; P<0.01 ). In conclusion, BE hastened luteal function but did not affect uterine involution.     

Uterine involution and ovarian changes during early post partum in autumn-lambing Corriedale ewes

The time of uterine involution and the changes in ovarian follicle populations were studied during early postpartum in multiparous, suckling Corriedale ewes lambing in the autumn. On Day 1 (n=5), Day 5 (n=4), Day 17 (n=4) and Day 30 (n=3) postpartum ewes underwent surgery to obtain ovaries and uteri. The weights of uteri and the lengths of the previous pregnant and nonpregnant horns were recorded as well as the presence of ovarian follicles larger than 1 mm in diameter. Uterine weight and length of uterine horns decreased (P2 to < 4 mm) follicles were also present. At days 17 and 30, aside from the small and medium follicles, all the ewes also had large (>/= 4 mm) follicles, and, at Day 30 2 ewes had large corpora lutea. We conclude that in autumn-lambing Corriedale ewes macroscopic uterine involution was complete around Day 17 post partum and that follicle development begins immediately after parturition, reaching preovulatory size before Day 17. In 2 of the 3 ewes studied until Day 30, ovulation (first progesterone increase) occurred after Day 17 (Days 18 and 25).     

Blood eicosanoids and immune indices during fasciolosis in water buffaloes

The effects of trickle infections of water buffaloes with Fasciola hepatica (60 metacercariae daily during a period of 20 days) on the blood plasma levels of prostaglandin E₂ (PGE₂), 6-keto-prostaglandin F₁ (6-keto-PG F₁) and thromboxane B₂ (TXB₂) were assessed. F. hepatica specific IgG and T- and B-lymphocyte ratios were evaluated as indicators of the immune response. Although the applied mode of infection did not result in clinical disease, changes in the plasma eicosanoid pattern were observed. Plasma PGE₂ values were significantly elevated in the infected water buffaloes 11 weeks post-infection (w.p.i.). In contrast, transiently but significantly lower TXB₂ values than in the uninfected controls were recorded in the phase of chronic fasciolosis. Plasma 6-keto-PGF₁ values were not considerably altered by the infection throughout the study period. F. hepatica-specific IgG were detected from 4 to 21 w.p.i. The proportion of peripheral T- and B-lymphocytes shifted towards B-cells from 2 to 12 w.p.i., gradually returning to control values afterwards. Although the water buffaloes appeared to be rather resistant to trickle infection with F. hepatica, moderate changes in plasma eicosanoid patterns were observed, indicating tissue damage and/or inflammation. Induction of the immune response could be monitored by an increase of F. hepatica-specific IgG, which was paralleled by a relative increase of the B-lymphocyte population.     

Genetic variants of ribosomal DNA and mitochondrial DNA between swamp and river buffaloes

To clarify the genetic relationship between Swamp and River buffaloes, the restriction fragment length polymorphisms (RFLPs) of nuclear genomic ribosomal DNA (rDNA) and cytoplasmic mitochondrial DNA (mtDNA) were analysed. Blood or liver samples from 73 Swamp and three River buffaloes were collected in East and South-east Asian countries. DNA samples from cattle, goats and sheep were used for comparisons. The analysis of rDNA allowed water buffaloes, cattle, goats and sheep to be characterized by four distinct repeat-types. However, swamp and river buffaloes showed the same repeat-type. Divergence of water buffalo and cattle is considered to have occurred approximately four to six million years ago. The RFLPs for mtDNA divided water buffaloes into three haplotypes, swamp-1, swamp-2 and river types. Swamp-1 accounted for 91% of all swamp buffaloes while swamp-2 was observed only in water buffaloes from Thailand (9%). All river buffaloes were of the same haplotype. No differences were observed between swamp and river buffaloes at the rDNA level. In contrast, a few distinct differences between them were found at the mtDNA level. Therefore, mtDNA polymorphisms provide an adequate means for classifying water buffaloes into either swamp or river buffaloes.     

Uterine involution, day and variance of first postpartum ovulation in mares treated with progesterone and estradiol-17beta for 1 or 2 days postpartum

The effects of a single or double regimen of exogenous progesterone and estradiol-17beta (P/E, total dose 300 mg P/20 mg E) were investigated in 50 postparturient Quarter Horse mares. In Trial 1, at 1 and 24 h after foaling, mares were injected with progesterone (150 mg) and estradiol-17beta (10 mg) (n = 7) or 0.9% NaCl (control, n = 13). In Trial 2, within 12 h after foaling, mares were injected with progesterone (300 mg) and estradiol-17beta (20 mg) (n = 13) or 0.9% NaCl (control, n = 17). Mares were examined daily by palpation per rectum and transrectal ultrasonography to determine the day of ovulation. The largest cross sectional diameters of each uterine horn and uterine body were measured ultrasonographically on Day 15 postpartum. Mean uterine diameters did not differ between treatment groups (P > 0.05) in Trial 1, Trial 2 or for combined data for both Trials 1 and 2. For mares bred on the first postpartum estrus pregnancy rates did not differ (P > 0.05) between treatment groups (16/18, 89%) and controls (22/30, 81%) nor was there a difference in mean day to first postpartum ovulation (P > 0.05) between treated and control groups in Trial 1, Trial 2 or Trials 1 and 2 combined. However, fewer (P < 0.05) total P/E treated mares (0/20) ovulated prior to Day 10 postpartum than did control mares (6/30). Variance in days to ovulation was lower (P < 0.05) for P/E treated mares (var = 3.73 days) than for control mares (var = 7.64 days) for data combined from Trials 1 and 2.     

Follicular waves during the oestrous cycle in Nili-Ravi buffaloes undergoing spontaneous and PGF2alpha-induced luteolysis

The objective of this study was to characterize the follicular waves and associated ovarian events during spontaneous and PGF(2alpha)-induced oestrous cycles in Nili-Ravi buffaloes. In Exp. 1, (n=13 oestrous cycles) follicular and luteal development was monitored by ultrasonography and jugular blood samples were collected simultaneously on alternate days. Of 12 oestrous cycles, 9 (75%) had two waves of follicular activity and only 3 (25%) had three waves. The mean (+/-S.E.M.) length of the oestrous cycle was shorter (P<0.05) in buffaloes with two waves than in those with three waves (21.2+/-0.1 days versus 22.8+/-0.1 days). In Exp. 2, follicular dynamics were compared in buffaloes undergoing spontaneous (n=12 oestrous cycles) and PGF(2alpha)-induced (n=9) regression of the corpus luteum (CL). The dynamics of ovulatory follicular growth during the 3 days before oestrus were similar (P>0.05) in buffaloes undergoing spontaneous and PGF(2alpha)-induced luteolysis. These results show that (1) the majority of buffaloes had a two wave pattern of follicular growth and emergence of a third wave was associated with a longer luteal phase, and (2) follicular dynamics during the 3 days before oestrus were similar in buffaloes undergoing spontaneous and PGF(2alpha)-induced luteolysis.     

Ovulation and fertility after PRID,PRID + GnRH and GnRH in anestrous buffaloes

Two experiments, one in the winter (January) and the other in the summer (June), were undertaken to treat anestrous buffaloes at a farm. During experiment I, animals were treated with a progesterone releasing intra-vaginal device (PRID)^a, whereas during experiment II, animals were allocated to PRID + GnRH^b (125 μg) and GnRH-treated (250 μg). In both the experiments, animals were retained as untreated controls. All the treated animals were clinically examined four days post-treatment and inseminated two or three times at 24-hour intervals. The ovulation and fertility rates of PRID, PRID + GnRH and GnRH-treated animals were 25 and 12 %, 43 and 14 %, and 11 and 0 %, respectively. None of the controls exhibited estrus during the course of the study.     

Effect of restricted suckling on ovarian activity and uterine involution in Brahman cows

Seventy-six Brahman cows and first-calf heifers ranging in age from three to five or more years were used to determine the effects of restricted suckling on postpartum ovarian activity and uterine involution. At 30 days postcalving, cows were randomly assigned to one of two groups: 1)normal suckling (34 cows) and 2)twice-daily suckling (42 cows). The cows were examined by rectal palpation weekly from parturition until the onset of the breeding season. The average interval from parturition to the presence of the first follicle larger than 10 mm (PPFI) was 36.0 +/- 1.0 days, and twice-daily suckling decreased PPFI length 8.1 days (P < 0.01); likewise, first- and second-calf heifers tended to have longer PPFI length (P < 0.05) than cows with three or more calvings. The average interval from parturition to first corpus luteum (PPCLI) was 59.0 +/- 2.0 days. PPCLI was affected by the age (P < 0.05) and weight of the cow at 30 days postpartum (P < 0.10). The average interval from parturition to first estrus (PPEI) was 68.0 +/- 5.0 days. PPEI was affected by suckling (P < 0.10) and month of parturition (P < 0.05). The average interval from parturition to uterine involution was 33.0 +/- 1.0 days. Uterine involution was influenced by month of calving (P < 0.01) and age of the cow (P < 0.05).     

The behaviour and welfare of buffaloes (Bubalus bubalis) in modern dairy enterprises

This review deals with the behaviour of river buffaloes (Bubalus bubalis), in confinement and in extensive conditions, also focusing on the effects of different housing and rearing conditions on their welfare. The behavioural repertoire expressed by buffaloes in extensive and intensive conditions is similar to those displayed by other domestic ruminants. However, through natural selection, buffaloes have also acquired several morphological, physiological and behavioural (i.e. wallowing) adaptations to hot climatic conditions. Buffaloes kept in intensive conditions and having no access to pasture and water for wallowing extend their periods of idling and are less often involved in investigative activities. Confinement is also associated with a reduction of space; however, no specific studies have been carried out to determine the specific requirements of this species. Space restriction can adversely affect various aspects of buffalo welfare, such as health (increased levels of lesions and injuries), social behaviour (increased number of agonistic interactions) and heat dissipation. The buffaloes, originating from tropical areas, are well adapted to large variations in food availability and quality, and to dietetic unbalances. As to human animal relationship, it has been observed that the incidence of stepping and kicking behaviour of buffaloes in the milking parlour is positively correlated with the frequency of oxytocin injections, whereas the frequency of positive stockperson interactions with the animals such as talking quietly, petting and gentle touching are negatively correlated with the number of kicks during milking. Data from farms where both dairy cattle and buffaloes are present show that avoidance distance measured in the pen is lower in buffaloes than in cattle. This may be attributed to the fact that buffaloes are generally recognised to be curious animals. Finally, the effects of different farming practices on animal-related indicators are described. However, these measures should be integrated into a monitoring protocol, such as the Welfare Quality® scheme, to reliably assess buffalo welfare in the current intensive farming conditions.     

Effect of month and season of calving on the length of subsequent calving interval in Nili-Ravi buffaloes

Data pertaining to records of 1,274 calving intervals for 400 breeding buffaloes maintained at the Livestock Experiment Station, Qadirabad, District Sahiwal (Pakistan), were analyzed to investigate the magnitude of influence of month and season of calving on the length of subsequent calving interval. The longest average calving interval of 609.9 days was recorded for buffaloes calving in January, while the shortest average calving interval of 502.8 days was observed for those calving in June, the difference being highly significant (P < 0.01). When the data were split in accordance with season of calving, the calving intervals for buffaloes calving in winter, spring, summer and autumn averaged 569.1, 570.6, 506.6 and 515.7 days, respectively. The differences between intervals for winter and spring calvings and between intervals for summer and autumn calvings were non-significant. All other differences were highly significant (P < 0.01). Thus, in most of the buffaloes calving in winter or spring, the first post-partum oestrus and conception are delayed till the following autumn or winter, resulting in significantly longer calving intervals. In contrast, animals calving in summer or autumn exhibit post-partum heat and conception during autumn or winter, resulting in shorter calving intervals.     

Factors affecting postpartum ovarian activity in crossbred primiparous tropical heifers

The relationship between postpartum ovarian activity and a total of 9 variables was studied in a dry tropical environment. Primiparous cows (n=61) that had shown no peripartum abnormalities, and were not suckled but milked twice daily, were used in the study. Independent variables included crossbreeding, sex of the calf, season, body condition, weight of cow at calving, age of dam at calving, uterine involution, calf weight and accumulated milk yield. Diet was a controlled variable. Dependent variables were first estrus postpartum and/or first milk progesterone elevation prior to first estrus. A bull fitted with a chin ball marker was used to detect first estrus postpartum, while ovarian structures were palpated per rectum once a week. Progesterone was measured by RIA in milk samples collected twice weekly. First postpartum estrus was detected at 56 +/- 32 days postpartum, a first milk progesterone elevation was observed in 50.8% of cows at 42 +/- 27 days. Cows calving in the dry season had longer intervals and those who calved males had shorter postpartum intervals. Accumulated milk yield affected both intervals significatly (p < 0.01). Weight, age and uterine involution were asociated with first milk progesterone elevation, while crossbreeding, weight at calving and weight postpartum change were associated with the dependent variables.     

Uterine epithelioid leiomyosarcoma in a pig-tailed macaque

Abstract: During ultrasonographic examination following spontaneous abortion, an approximately 6-cm diameter mass of mixed echogenicity is detected in the uterine wall of a mature pig-tailed macaque (Macaca nemestrina). Grossly, the mass is associated with the uterine fundus, multilobulate and tan with red-black mottled foci on cut surface. Microscopically, the invasive mass is composed of poorly differentiated round cells arranged in sheets and bundles supported by a fine fibrous stroma. On immunohistochemical evaluation, neoplastic cells contain vimentin, desmin, muscle actin, and smooth muscle α-actin antigens and do not react with antibodies to low and high molecular-weight cytokeratins. Cell morphology and immunohistochemical results indicate a diagnosis of epithelioid leiomyosarcoma (clear cell variant).     

Studies on maturation of genitalia in murrah buffalo-heifers

Studies on the growth of uterine horns and cervix were conducted on 161 prepuberal, puberal and postpuberal (non-gravid) farm-reared buffalo heifers. Age of the buffalo heifers varied between 17 and 37 months, while body weight range was 180–500 kg. The approximate diameters of the mid-cervix and uterine cornua (at the point of pseudobifurcation) were visualized mentally by palpating the organs per rectum with fingers of the left hand while focusing the adjudged diameters between the jaws of a vernier caliper held in the other hand. Both uterus and cervix increased progessively with the advancing age and body weight of heifers, however, the growth of the cervix was more consistent compared to that of the uterus. A marked enlargement in uterine cornua was noted at the time of puberty, however, the rate of cornual growth declined with subsequent estrous cycles.