Evaluation of cytokine expression by blood monocytes of lactating Holstein cows with or without postpartum uterine disease

Whereas neutrophils are the main phagocytic leukocytes, monocytes and macrophages are actively involved in immunomodulation after infection. Recent studies have demonstrated that neutrophil function is impaired by the state of negative energy balance around parturition, and that cows that develop uterine disease have a greater degree of negative energy balance than healthy cows. The objectives of this study were to compare monocyte gene expression and protein secretion of selected cytokines from calving to 42 d after calving in Holstein cows that did or did not develop uterine disease. Real time quantitative RT-PCR (Tumor necrosis factor-α (TNFα), Interleukin (IL)-1β, IL-6, IL-8 and IL-10) and ELISA (TNFα, IL-1β and IL-8) were used to evaluate cytokine response following in vitro stimulation of blood-derived monocytes with irradiated E. coli. Relative to unstimulated cells, E. coli-stimulated monocytes from cows with metritis had lower gene expression of key pro-inflammatory cytokines than healthy cows from calving to 14 d after calving (TNFα at 0, 7, and 14 d after calving, IL-1β and IL-6 at 7 and 14 d after calving; P < 0.05). There were no significant differences between groups for expression of IL-8 or the anti-inflammatory cytokine IL-10. This was due, in part, to higher gene expression in unstimulated monocytes (TNFα, IL-1β, IL-6 and IL-10) in early lactation from cows with metritis. Expression of mRNA in stimulated cells (relative to housekeeping genes) was lower for TNFα (7 and 14 d postpartum) and for IL-10 (7 and 14 d postpartum) in cows with metritis. Concentration of TNFα was lower in the culture medium of E. coli-stimulated monocytes from cows with metritis than healthy cows at calving and 7 and 21 d after calving (P < 0.05). Circulating cytokine concentrations were not different between groups for IL-8 and were below the limits of detection for TNFα and IL-1β. Cytokine gene expression and production were similar between healthy cows and cows that developed endometritis, diagnosed cytologically at 42 d after calving. We concluded that altered levels of expression and production of pro-inflammatory cytokines postpartum could contribute to impaired inflammatory response and predispose cows to development of metritis.     

Effect of postpartum suppression of ovulation on uterine involution in dairy cows

The objective of this study was to investigate the effect of time of first postpartum ovulation after calving on uterine involution in dairy cows with and without uterine puerperal disease. Transvaginal follicular puncture (FP) of follicles >6 mm suppressed ovulation and development of a CL until Day 42 after calving. Fifty-three lactating Holstein Friesian cows (3.4 ± 1.2 years old, parity 2.5 ± 1.0 [median ± mean absolute deviation]) were divided into groups on the basis of the presence (UD+) or absence (UD−) of uterine disease and whether FP was carried out (FP+) or not (FP−). Uterine disease was defined as the occurrence of retained fetal membranes and/or metritis. This resulted in the following groups: UD−FP− (n = 15), UD−FP+ (n = 13), UD+FP− (n = 13), and UD+FP+ (n = 12). A general examination, vaginoscopy, transrectal palpation, and transrectal B-mode sonography of the reproductive organs were conducted on Days 8, 11, 18, and 25 and then every 10 days until Day 65 after calving. After hormonal synchronization of ovulation (cloprostenol between Days 55 and 60 postpartum and GnRH 2 days later), cows were inseminated in the next spontaneous estrus. On average, the cows ovulated on Day 21.0 ± 6.0 (UD−FP−), 50.0 ± 4.0 (UD−FP+), 16.0 ± 3.0 (UD+FP−), and 48.0 ± 2.0 (UD+FP+) postpartum. Calving-to-conception interval and first-service conception rates were not affected by FP (P > 0.05). Healthy cows with FP had smaller (P < 0.05) uterine horn and cervical diameters assessed sonographically than cows without FP. FP reduced the prevalence of purulent vaginal discharge and uterine size assessed transrectally in UD+ cows (P < 0.05). The results showed that suppression of an early ovulation by transvaginal FP improved uterine involution in cows with and without uterine disease.     

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.     

Influence of bull biostimulation, season and parity on resumption of ovarian activity of zebu (Bos indicus) cattle following parturition

A total of 135 postpartum suckled zebu cows were assigned randomly at calving to two treatments: cows exposed to vasectomised bulls (Mature Bull Exposure, MBE) and cows not exposed to bulls (No Bull Exposure, NBE) for a period of 150 days postpartum. This study was conducted to determine the influence of bull biostimulation, season and parity on postpartum reproductive performance of Bos indicus cattle. The trials were conducted in two seasons: cows calving in the dry season and cows calving in the rainy season. Cows with an increase in serum progesterone (P(4)) concentration of >1 ng/ml from the weekly blood samples were used to analyse the number of days from calving to the time of resumption of ovarian activity. The introduction of vasectomised bulls shortened the postpartum anoestrus in cattle following calving. Mean interval from calving to resumption of ovarian activity for the MBE cows was 71.7 days, which was significantly earlier following calving than the NBE cows with a duration of 77.8 days. By 60 to 80 days postpartum, the proportion of cows at resumption of postpartum activity for MBE cows was greater than for the NBE cows.Mean interval from calving to resumption of ovarian activity for cows that calved in the dry season was 71.3 days, which was significantly earlier than for cows that calved in the rainy season (78.6 days). At 60 days postpartum, the proportion of cows at resumption of postpartum ovarin activity for cows that calved in the dry season was greater than the cows that calved in the rainy season. Mean interval from parturition to resumption of ovarian activity for cows with three to five calvings was 65.1 days, which was significantly earlier than the value of 71. 2 days for cows with one to two calvings. By 60 to 80 days postpartum, the proportion of cows at onset of ovarian activity for cows with three to five calvings was greater than those cows with one to two calvings. Cows that calved in the dry season completed uterine involution by 24.4 days, which was significantly shorter than the duration of 26.5 days for cows that calved in the rainy season. Cows with three to five calvings completed uterine involution earlier than those with one to two calvings. It is concluded that bull-cow biostimulation influences reproductive activity in the cow possibly through olfactory cues (pheromones).     

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).     

Peripheral blood leukocytes of cows with subclinical endometritis show an altered cellular composition and gene expression

Subclinical endometritis (SCE) is an important postpartum disease in dairy cows, but conventional cytobrush diagnosis often gives imprecise results. The aim of this study was to analyze disease-associated changes in peripheral blood as potential diagnostic parameters. Cellular subpopulations of blood leukocytes from cows with or without SCE (45–55 days postpartum) were flow-cytometrically quantified. Gene expression of whole blood leukocytes was assessed by PAXgene analysis. Subclinical endometritis cows showed significantly higher number of blood mononuclear cells and neutrophils. Among mononuclear cells, numbers of B-cells, NK-cells, and CD172a-positive monocytes were significantly elevated. Compared with non-SCE cows, blood leukocytes of SCE cows significantly expressed higher copy numbers of CXCL8, TNF, and IL12. To test whether circulating plasma factors are responsible for these changes, leukocytes, polymorphonuclear cells, and monocyte subpopulations (classical, intermediate, nonclassical) of healthy cows were stimulated with plasma of SCE and non-SCE cows. Although gene expression of whole leukocytes and polymorphonuclear cells remained unaltered, plasma from SCE animals significantly elevated expressed messenger RNA copy numbers of CXCL8, CXCL1, and IL1B in intermediate monocytes. In conclusion, elevated number of selected mononuclear subpopulations in peripheral blood and enhanced expression of distinct genes encoding for inflammatory mediators in blood leukocytes reflect the subclinical uterine inflammatory process in cows. Whether the observed changes in the periphery of SCE cows are the consequence of the uterine inflammatory process, or whether they affect the pathogenesis of the disease is currently unknown.     

Bacteriological investigation of the postpartum uterus: Relationship to involution and histopathological findings

A bacteriological investigation of the postpartum uteri of suckled and nonsuckled Bunaji (White Fulani) cows was undertaken to determine the microflora, the effect of suckling on the flora and the influence of the flora on uterine involution and histopathology. Uterine contamination by bacteria was highest between days 10 and 21 postpartum with Staphylococcus aureus being the commonest organism isolated. There was no consistent isolation of one type of bacterial organism from a cow throughout the sampling period. Suckling did not affect the incidence of uterine contamination by bacteria. Foci of leucocytes (mainly lymphocytes) were found in uterine sections in an increasing frequency with days postpartum. Bacteria were isolated from only 3 of the 12 uteri that had the leucocytic foci. Uterine involution was normal and complete by 25 days after calving irrespective of the presence or absence of bacteria or leucocytic foci. We concluded that postpartum uterine contamination by bacteria was not influenced by suckling and that puerperal uterine contamination by bacteria did not interfere with the normal process and duration of uterine involution.     

Screening for high fertility in high-producing dairy cows

A retrospective study involving 2756 pregnancies from two commercial dairy herds in northeastern Spain determined relationships between management, production and reproductive data, and high fertility (conception before 90 days in milk) in high-producing dairy cows. High fertility was registered in 989 (35.9%) cows. The following data were recorded for each animal: herd, repeated animal (cows included two or more times within the study in which data were obtained from different lactational periods), parity (primiparous versus multiparous), previous twinning, reproductive disorders following calving (retained placenta, primary metritis) and at postpartum gynecological examination (incomplete uterine involution, pyometra and ovarian cysts), days in milk at conception, previous estrous synchronization and season of calving and conception. In order to evaluate the possible effect of high production during the peak milk yield on subsequent fertility, daily milk production at Day 50 postpartum was also recorded and cows were classified as high (> or = 50 kg) and low (< 50 kg) producers. Logistic regression analysis indicated no significant effects of herd, repeated animal, previous twinning, reproductive disorders such as primary metritis, incomplete uterine involution, pyometra and ovarian cysts, previous estrous synchronization and season of calving and insemination. Based on the odds ratio, the likelihood of high fertility increased in high-producer cows by a factor of 6.8. High fertility was less likely for multiparous cows (by a factor of 0.35) and for cows suffering placenta retention (by a factor of 0.65). High fertile cows produced a mean of 49.5 kg milk at Day 50 postpartum, in contrast to that 43.2 kg milk of the remainder cows. These findings question the negative effect of high production on fertility. Our results indicated that high individual cow milk production can be positively related to high fertility.     

Effect of Feeding on Reproductive Performance in Finnish Dairy Cows

The effect of 2 types of feeding on the involution of the genital tract and on the fertility was studied in 79 Finnish dairy cows on a research farm. The cows were fed twice a day with home-produced feed in accordance with the Finnish feeding standard. The cows were divided according to the type of feed into a hay-urea group and a silage group. The cows were examined clinically by rectal palpation 3 times a week during 8 weeks postpartum. The time required for the complete involution of uterus and cervix was recorded. The cows in the silage group had a significantly longer time in uterine involution, a lower fertility rate at first insemination and a longer interval from calving to conception than those in the hay-urea group. The pregnancy rate in the first insemination was in the hay-urea group 91 % and in the silage group 57 %. The interval between calving and the first insemination was shorter in the hay-urea group than in the silage group; 71.4 ± 10.6 and 74.0 ± 10.3 days, respectively. The interval between calving and conception was in the hay-urea group significantly (p < 0.01) shorter than in the silage group; 74.8 ± 15.1 and 89.5 ± 24.4 days, respectively.     

The effect of treatment with a GnRH analogue on postpartum reproductive performance in Friesian cows

Three trials were conducted to examine the effect of GnRH injection on the reproductive performance of Friesian cows. In the first trial 100 μg gonadotrophin releasing hormone synthetic analogue (GnRH) was given at the time of A.I. to 32 cows while a control group received a saline placebo injection. Conception rate to first insemination was significantly higher in treated than control cows (81.3 v. 54.8%, P < 0.05).In the second trial, two groups of 19 cows each received either 100 μg GnRH or saline at 15 days postpartum. A total of 60 cows was used in the third trial in which GnRH was given at either 50 or 100 μg dose level on either days 7 or 15 postpartum while control cows were untreated. In both trials GnRH treatment reduced the intervals from calving to complete uterine involution, first ovulation and first detected oestrus. The effect was most pronounced when 100 μg GnRH was given on day 7 postpartum. Number of days open and number of services per conception were appreciably reduced when 100 μg GnRH was given at either 7 or 15 days postpartum with maximal effect when given 15 days after calving.     

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.     

Time-dependent mRNA expression of selected pro-inflammatory factors in the endometrium of primiparous cows postpartum

Background  

Inflammatory processes and infections of the uterine wall must be accepted as a physiological event in dairy cows after calving. This might result in clinical or subclinical endometritis which is assumed to impair reproductive performance in the current lactation. Several cytokines and acute phase proteins have been discussed as local and systemic mediators of these inflammatory processes. The aim of the present study was to investigate the endometrial mRNA expression of the chemokine CXC ligand 5 (CXCL5), interleukin 1β (IL1B), IL6, IL8, tumour necrosis factor alpha (TNF), prostaglandin-endoperoxide synthase 2 (PTGS2) and haptoglobin (HP) in the postpartum period.     

Resumption of postpartum ovarian cyclicity in high-producing Holstein cows

The objective of this study was to investigate the resumption of ovarian cyclicity postpartum in high-producing dairy cows in commercial dairy farms under subtropical conditions. The cows were kept in a free-stall or tie-stall barn. Milk samples were collected from cows twice weekly, and progesterone in the skim milk was assayed by double-antibody ELISA. Cows were examined rectally and vaginoscopically at 2-week intervals after calving. Body condition score (BCS) and body weights were taken before and after calving. A cow was considered to have resumed ovarian cyclicity on the day of ovulation if followed by regular ovarian cycles. Thirty seven percent (n=20/54) of the cows had normal resumption of ovarian cyclicity (resumption within 45 days after calving), and 63% (n=34/54) had delayed resumption (resumption did not occur until >45 days after calving). Delayed resumption Type I (one or more ovarian cycles with luteal phase >20 days, i.e. prolonged luteal phase; 31.5%) and delayed resumption Type II (first ovulation did not occur until > or =45 days after calving, i.e. delayed first ovulation; 24.1%) were the most common types of delayed resumptions. Almost half (46.3%) of the cows did not resume their ovarian cyclicity until >65 days postpartum. Cows with delayed resumption Type I had a higher incidence of abnormal cervico-vaginal discharge (64.7%) and incomplete uterine involution (94.1%) compared to cows with normal resumption (P<0.01). The BCS of cows with delayed resumption Type II were lower than those of normal resumption cows at 5 weeks and later in the postpartum period (P<0.05). Approximately two-thirds of high-producing cows had delayed resumption of ovarian cyclicity postpartum. Prolonged luteal phase and delayed first ovulation were two important ovarian dysfunctions that delayed postpartum resumption of cyclicity in high-producing dairy cows.     

Ultrasonic observation of postpartum uterine involution in the cow

To learn about the progress of postpartum uterine involution in the dairy cow, the authors wanted to know whether the ultrasonic linear scanner is applicable for this purpose. The uterus was observed by the transducer, which was inserted into the rectum. Clear cross-sectional images of the endometrium were obtained in this way; however, those of the myometrium and the perimetrium were not very clear. Observation on the progress of uterine involution was started on Day 8 postpartum and continued to Day 43 postpartum. The diameter and area of the uterine horn and the endometrium in cross-sectional ultrasonograph images of the endometrium were estimated using a Magiscan 2 image analyzer. The relationship between the estimated dimensions of the uterine horn and postpartum days were satisfactorily fitted into polynomial regressions. Using the ultrasonic linear scanner, it was confirmed that uterine involution was completed at approximately 40 d postpartum. The applicability of the ultrasonic linear scanner for diagnosis of uterine involution in the postpartum cow is demonstrated by the results of the present study.     

Effects of gonadotropin releasing hormone and prostaglandin F(2)alpha on the reproductive performance in postpartum cows

A study was conducted to determine whether treatment with gonadotropin releasing hormone (GnRH) in combination with prostaglandin F(2)alpha (PGF(2)alpha) could enhance ovarian activity and uterine involution in postpartum dairy cows to reduce the calving interval. Cows were randomly assigned to one of three treatment groups. Cows (n = 8) in Group 1 received 100 mug GnRH intramuscularly (i.m.) twice on Day 20 and Day 35 postpartum, and 25 mg PGF(2)alpha i.m. on Day 47 postpartum. Group 2 (n = 8) received a single i.m. injection of 100 mug GnRH on Day 25 postpartum and 25 mg PGF(2)alpha i.m. on Day 37 postpartum. The Control Group (n = 9) did not receive hormonal treatment. Palpation per rectum of the reproductive organs and serum progesterone (P) determination were performed twice a week to monitor ovarian activity and uterine involution. Postpartum interval to the first ovulation was short in treated groups (Group 1, 21.0 d; Group 2, 26.3 d) compared with Control Group (30.1 d, P < 0.05). Likewise, mean frequency of ovulation was increased in both treated groups compared with the Control Group (P < 0.05). Cows in treated groups required fewer days to complete uterine involution than in the Control Group. The mean interval to the first service, the conception rate at first service and the number of services per conception showed no significant differences among the three groups, but the mean days from calving to conception were shorter for the treated groups (78.7 d in Group 1; 83.3 d in Group 2) than (109.1 d, P < 0.05) for the Control Group. Our results suggest that combined treatment with GnRH and PGF(2)alpha may enhance ovarian activity in the postpartum cow, resulting in improved reproductive performance.     

Assessment of Early Postpartum Reproductive Performance in Two High Producing Estonian Dairy Herds

Early postpartum (6 weeks) ovarian activity, hormonal profiles, uterine involution, uterine infections, serum electrolytes, glucose, milk acetoacetate and blood urea nitrogen (BUN) levels were studied in 2 Estonian high producing dairy herd with annual milk production of 7688 (Farm A) and 9425 (Farm B). From each farm 10 cows, with normal calving performance were used. Blood samples for the hormonal (PGF_2α-metabolite, progesterone) analyses were withdrawn. On day 25 PP blood serum samples were taken for the evaluation of metabolic/electrolyte status. On the same day estimation of milk acetoacetate values was done. The ultrasound (US) was started on day 7 PP and was performed every 3^rd day until the end of experiment. Uterine content, follicular activity and sizes of the largest follicle and corpus luteum were monitored and measured. Vaginal discharge and uterine tone were recorded during the rectal palpation. Each animal in the study was sampled for bacteriological examination using endometrial biopsies once a week. Two types of PGF_2α-metabolite patterns were detected: elevated levels during 14 days PP, then decline to the basal level and then a second small elevation at the time of final elimination of the bacteria from the uterus; or elevated levels during first 7 days PP, then decline to the basal level and a second small elevation before the final elimination of bacteria. Endometritis was diagnosed in 5 cows in farm A and in 3 cows in farm B respectively. In farm A, 5 cows out of 10 ovulated during experimental period and in 1 cow cystic ovaries were found. In farm B, 3 cows out of 10 ovulated. In 3 cows cystic ovaries were found. Altogether 40% of cows had their first ovulation during the experimental period. Three cows in farm A and 5 cows in farm B were totally bacteria negative during the experimental period. The most frequent bacteria found were A. pyogenes, Streptococcus spp., E. coli., F. necrophorum and Bacteroides spp. The highest incidence of bacteriological species was found during the first 3 weeks in both farms. All animals were free from bacteria after 5^th week PP in farm A and after 4^th week in farm B respectively. Serum electrolytes and glucose levels were found to be within the reference limits for the cows in both farms. No significant difference was found between farms (p > 0.05). Low phosphorus levels were found in both farms. Significant difference (p < 0.05) was found in BUN levels between farms. In both farms milk acetoacetate values were staying within the reference range given for the used test (<100 μmol/l). The uterine involution and bacterial elimination in the investigated cows could consider as normal but more profound metabolic studies could be needed to find reasons for later resumption of ovarian activity. Some recommendations to changing feeding regimes and strategies should also be given.     

Uterine Microbiota Progression from Calving until Establishment of Metritis in Dairy Cows

The objective of this study was to evaluate the progression of the uterine microbiota from calving until establishment of metritis. Uterine swabs (n = 72) collected at 0, 2, and 6 ± 2 days postpartum (dpp) from 12 metritic and 12 healthy cows were used for metagenomic sequencing of the 16S rRNA gene on the Illumina MiSeq platform. A heat map showed that uterine microbiota was established at calving. The microbiota changed rapidly from 0 to 6 ± 2 dpp, with a decrease in the abundance of Proteobacteria and an increase in the abundance of Bacteroidetes and Fusobacteria, which were dominant in metritic cows. Uterine microbiota composition was shared; however, metritic and healthy cows could be discriminated using relative abundance of bacterial genera at 0, 2, and 6 ± 2 dpp. Bacteroides was the main genus associated with metritis because it was the only genus that showed significantly greater abundance in cows with metritis. As the abundance of Bacteroides organisms increased, the uterine discharge score, a measure of uterine health, worsened. Fusobacterium was also an important genus associated with metritis because Fusobacterium abundance increased as Bacteroides abundance increased and the uterine discharge score worsened as the abundance increased. The correlation with uterine discharge score and the correlation with Bacteroides or Fusobacterium showed that other bacteria, such as Helcoccocus, Filifactor, and Porphyromonas, were also associated with metritis. There were also bacteria associated with uterine health, such as “Candidatus Blochmannia,” Escherichia, Sneathia, and Pedobacter.     

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.     

Relationship between intra-uterine bacterial contamination, endotoxin levels and the development of endometritis in postpartum cows with dystocia or retained placenta

A study was conducted to investigate the relationship between intra-uterine bacterial contamination, endotoxin levels and the development of endometritis in cows that experienced a dystocia or retained their placenta. Fifteen healthy cows, 31 cows with retained placenta (RP) and 13 cows that had dystocia were clinically examined 1 or 2 days after parturition when a uterine swab for bacteriological examination was taken. In addition, plasma and uterine lochia samples were collected to determine lipopolysaccharide (LPS) and the plasma IgG anti-LPS concentrations. Subsequently, 15 RP and 6 dystocia cows were initially left untreated and another uterine swab was collected at 2 and 4 wk postpartum. Immediately after calving, RP cows had significantly higher LPS levels in uterine lochia (average of 2.24 x 10(4) Endotoxin Units (EU)/mL) as compared to dystocia and healthy postpartum cows (average of 0.10 and 0.26 EU/mL, respectively). However, plasma LPS levels were below the detection limit (<0.036 EU/mL platelet-rich plasma) in all groups of cows. IgG anti-LPS levels in plasma were not significantly different between the 3 groups immediately postpartum (average of 26, 16 and 44 Median Units (MU)/mL) for healthy, dystocia and RP cows, respectively), but they were significantly lower when compared to plasma IgG anti-LPS levels of healthy cows at more than 2 months postpartum (mean 83 MU/mL). High LPS levels in lochia at 1 or 2 days postpartum were significantly related to abnormal cervical discharge, the presence of Escherichia coli, black pigmented gram-negative anaerobes and Clostridium spp. shortly after calving, and Arcanobacterium pyogenes and gram-negative anaerobes in the uterus at 14 days postpartum. These results suggest that the presence of E. coli and LPS (endotoxins) in lochia early postpartum favor the development of uterine infections by A. pyogenes and gram-negative anaerobes later postpartum. LPS were not observed in plasma, suggesting that either they are not absorbed into the blood, or they are efficiently detoxified by IgG anti-LPS or other detoxification mechanisms.