Postpartum uterine flora following normal and abnormal puerperium in cows

doi:10.1016/0093-691X(90)90522-U

Theriogenology

Volume 34, Issue 2, August 1990, Pages 291-302

https://doi.org/10.1016/0093-691X(90)90522-U Get rights and content

Abstract

Uterine swabs were taken from the uteri of three groups of cows (n = 5 each) on Days 1 and 2 and then at 3-d intervals up to 32 d post partum. Group I cows had a normal delivery, Group II cows had dexamethasone-induced parturition, and Group III cows were subjected to a Caesarian section. All cows in Group III and two cows in Group II retained their fetal membranes. A total of 230 bacterial isolates were made, and the most frequently isolated bacteria were $Escherichia$ $coli$ , streptococci, $Proteus$ $spp.$ , and clostridia. The highest number of bacterial isolates and the longest mean time to complete uterine involution were in the Group III cows.

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The incidence of bovine endometritis, which has a negative impact on the reproduction of dairy cows, has been recently increasing. In this study, the differential markers and metabolites of healthy cows and cows with endometritis were analyzed by measuring blood biochemical indicators and immune factors using biochemical and enzyme-linked immunosorbent assay kits combined with nontargeted metabolomics. The LC-QTOF platform was used to evaluate the serum metabolomics of healthy cows and cows with endometritis after 21–27 days of calving. The results showed that glucose, free fatty acid, calcium, sodium, albumin, and alanine aminotransferase levels were significantly lower in the serum of cows with endometritis than in healthy cows (P < 0.05). However, the serum potassium, interleukin-1, interleukin-6, and tumor necrosis factor levels were significantly higher in cows with endometritis (P < 0.05). In addition, the serum metabolome data analysis of the two groups showed that the expression of 468 metabolites was significantly different (P < 0.05), of which 291 were upregulated and 177 were downregulated. These metabolites were involved in 78 metabolic pathways, including amino acid, nucleotide, carbohydrate, lipid, and vitamin metabolism pathways; signal transduction pathways, and other biological pathways. Taken together, negative energy balance and immune activation, which are related to local abnormalities in amino acid, lipid, and carbohydrate metabolism, were the important causes of endometritis in dairy cows. Metabolites such as glucose, carnosine, dehydroascorbic acid, L-malic acid, tetrahydrofolic acid, and UDP-glucose may be used as key indicators in the hematological diagnosis and treatment of endometritis in dairy cows.
Characterization of cytokine gene expression in uterine cytobrush samples of non-endometritic versus endometritic postpartum dairy cows
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To better understand uterine inflammation in postpartum dairy cows we collected sequential cytobrush samples at 29–35 and at 49–55 d in milk (DIM). Based on the uterine cytology, cows were classified as Non-endometritic (n = 23; <18% neutrophils) or Endometritic (n = 12; ≥18% neutrophils) at 29–35 DIM and Non-endometritic (n = 17; <10% neutrophils) or Endometritic (n = 9; ≥10% neutrophils) at 49–55 DIM. Cows defined as Sham Controls (n = 6) were examined by vaginoscopy at 29–35 DIM and identified as Non-endometritic (<10% neutrophils) at 49–55 DIM. Cytokine gene expression in cytobrush samples was assessed using qRT-PCR. Sham Controls did not differ significantly (P > 0.17) from Non-endometritic cows at 49–55 DIM and these data were combined (n = 23). Uterine cytology-based classification using the aforementioned thresholds effectively separated cows into groups with Endometritic cows having significantly higher expression of pro-inflammatory (interleukin (IL)-1α, IL-1β, IL-6, IL-8, IL-17A CSF-1; P < 0.01) and regulatory (IL-1RA and IL-10; P < 0.03) cytokines, relative to Non-endometritic cows. Furthermore, Non-endometritic cows showed a significant decline (P < 0.03) in the expression of pro-inflammatory (IL-1α, IL-6, IL-8) and regulatory (IL-10) cytokine genes as the postpartum period progressed; whereas Endometritic cows exhibited a sustained elevation in transcript abundance throughout the sample period for both pro-inflammatory and regulatory cytokine genes. Expression of transforming growth factor (TGF) genes was more complex with TGF-β3 expression significantly (P < 0.01) lower at 29–35 DIM and TGF-β1 gene expression significantly (P < 0.03) increased at 49–55 DIM in Endometritic versus Non-endometritic cows. Expression of TGF-β2 gene was 2.7-fold higher (P < 0.01) at 29–35 DIM in cows that remained Endometritic when compared to cows recovering by 49–55 DIM. Some Non-endometritic cows (n = 4) at 29–35 DIM were reclassified as Endometritic at 49–55 DIM. The sampling procedures at 29–35 DIM did not alter either the cellular response (P > 0.43) or cytokine gene expression (P > 0.17) at 49–55 DIM. In conclusion, normal uterine involution is characterized by a progressive decline in pro-inflammatory and regulatory cytokine gene expression, while cows with endometritis show a dysregulated inflammatory process characterized by a sustained elevation in pro-inflammatory and regulatory cytokine gene expression. This analysis also shows that decreased TGF-β2 gene expression at 29–35 DIM may be an indicator of recovery from endometritis.
Prevalence and antibiotic resistance pattern of certain types of bacterial flora in uterine ewe's samples
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Citation Excerpt :
Several studies have been made in Iraq which focused on isolation of certain microorganisms behaving as an opportunistic bacteria in uteri of different animals [14–16]. Many researchers indicated that the reproductive system contains normal flora [17–21]. Beta-lactamases are enzymes that are major causes of bacterial resistance to the beta-lactam family of antibiotics such as penicillins, cephalosporins, cephamycins, and carbapenems.
A study was carried out in abattoirs to identify certain bacterial flora in a number of samples collected from ewes' uteri with detection of phenotypic and genotypic antibiotic resistance patterns of the identified bacteria. The study was done in the South of Iraq during a period that started in February and ended in March 2015. Seventy-nine samples were collected randomly and aseptically, examined grossly, cultured using standard bacteriological techniques and examined for antibiotic resistance. Thirty-one isolates were reported belong to the following bacteria with resistance percentages accordingly: Escherichia coli 41. 94% (No: 13), Klebsiella spp. 29.03% (No: 9), Enterobacter spp. 16.13% (No: 5), Pseudomonas aeruginosa 6.45% (No: 2) and Proteus spp. 6.45% (No: 2). Results revealed that 100% (No: 31) of isolates were resistant to oxacillin while resistance to both ampicillin and tetracycline appeared in 93.64% (No: 30), 41.92% (No: 13) of isolates respectively, moreover there was for some extent resistance to ceftriaxone 9.68% (No: 3), while the isolates were highly susceptible to cefamandole and gentamicin. The isolates were also examined to determine the presence of bla_SHV genes by PCR assay which showed that 12.9% of isolates harbored this gene. This study contributes to a better knowledge about identified bacterial species inhabiting the uterus of ewes and exerting a significant and distinct antimicrobial resistance pattern.
Mucosal immunity of the postpartum bovine genital tract
2017, Theriogenology
Citation Excerpt :
This observation suggests that, at least in a subpopulation of cows, sustained infection and prolonged uterine inflammation may be attributed primarily to a failure in uterine defenses and not to specific bacterial pathogens. Bacterial species that have been recognized as non-specific uterine pathogens include E. coli, T. pyogenes, Fusobacterium (F.) necrophorum and Prevotella (P.) melaninogenicus [81–87]. Among these, E. coli is the predominant non-specific uterine pathogen associated with bovine metritis during the first wk postpartum [81–84].
In this review, mucosal immune defense mechanisms used to control infections in the bovine genital tract (vestibule, vagina, cervix, uterus and oviduct) during the postpartum period are reviewed. Knowledge gaps are highlighted to emphasize the need for further investigations. Physical barriers to the entry of microbes include vulvar sealing, vestibule-vaginal constriction, a narrow cervical opening and the mucosal epithelium along with the overlying mucus layer. Genital tract mucosal epithelial cells recognize damage-associated molecular patterns and pathogen-associated molecular patterns and respond by secreting antimicrobial peptides and cytokines to recruit and activate immune cells. Neutrophils and macrophages represent the first line of innate immune defenses recruited by cytokines to the site of inflammation. Macrophages, endometrial epithelial cells and dendritic cells interact with T-cells to elicit cellular responses and regulate antibody responses. Immune regulatory components such as M2-macrophages and regulatory T-cells, although less studied, may work in conjunction with epithelial cell regeneration to coordinate involution of the postpartum uterus and prepare the genital tract for the next pregnancy. A role for the vaginal and uterine microbiome in modulating uterine inflammation is an emerging research focus and further studies are required to integrate information on the nutritional and metabolic status of cows with innate immune responses and host-microbiome interactions. A greater understanding of these complex interactions is critical for developing more effective therapies for the prevention and treatment of uterine inflammation.
Subclinical endometritis in beef cattle in early and late postpartum: Cytology, bacteriology, haptoglobin and test strip efficiency to evaluate the evolution of the disease
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In cows, detrimental effects on fertility are mainly caused by clinical and subclinical endometritis (SEM). As demonstrated in previous work, Piedmontese cattle are affected by a higher rate of infertility and presence of SEM. The objective of this study is to assess the pattern of SEM at 30 and 60 days postpartum by evaluating the correlation between uterine cytology and microbiology, analyzing SEM consequences on reproductive career and verifying the reliability of rising inflammatory proteins - haptoglobin and the test strip test. Seventy healthy cows were enrolled and sampled at 30 and 60 days postpartum; cytology and bacteriology as well as haptoglobin and test strip were evaluated. The ROC curve for cytology set the optimal cut-off at 6.5% at 30 days and 2.5% at 60 days for a Partum-to-Conception (PC) interval of 120 days. The cytological positivity was negatively correlated with fertility, at 30 days, but not at 60 days. A positive bacteriological test was not correlated with an increase in the PC at either 30 or 60 days postpartum. The presence of a calving parlor affect the fertility (P < 0.05) but not the presence of parity or suckling calf and parity. The ROC curve for strip test protein at 30 days postpartum set a cut-off of 2% for PC. No difference in serum haptoglobin was observed between negative or positive cytology/bacteriology in postpartum cattle. The test strip results for proteins have demonstrated a utility at 30 days postpartum for screening the cows that are at risk of developing an increased PC > 120 days.
Dynamics of the microbiota found in the vaginas of dairy cows during the transition period: Associations with uterine diseases and reproductive outcome
2017, Journal of Dairy Science
We investigated the microbiota found in the vaginas of Holstein dairy cows during the transition period and described the differences in bacterial composition and total bacterial load (TBL) associated with disease and fertility. Vaginal swabs were collected at −7, 0, 3, and 7 d relative to parturition from 111 dairy cows housed on a commercial dairy farm near Ithaca, New York. Microbiota were characterized by next-generation DNA sequencing of the bacterial 16S rRNA gene, and TBL was determined by real-time quantitative PCR. We applied repeated-measures ANOVA to evaluate the associations of uterine disease and related risk factors with the microbiota and TBL. We estimated phylum-specific bacterial load by multiplying the TBL by the relative abundance of each phylum observed in the metagenomics results. We confirmed the validity of this approach for estimating bacterial load by enumerating the number of bacteria in an artificial sample mixed in vitro and in clinical and healthy vaginal samples. Phyla associated with uterine disease and related risk factors were Proteobacteria, Fusobacteria, and Bacteroidetes. Cows with retained placenta and healthy cows had similar TBL at the day of parturition, but at d 7 postpartum, cows with retained placenta showed a significantly higher TBL, mainly driven by higher estimated loads of Fusobacteria and Bacteroidetes. Cows diagnosed with metritis had a significantly higher estimated load of Proteobacteria at d −7 and at calving and higher estimated loads of Fusobacteria in the postpartum samples. Additionally, the estimated load of Bacteroidetes at d 7 postpartum was higher for cows diagnosed with endometritis at 35 days in milk. Higher estimated loads of Fusobacteria and Bacteroidetes were also evident in cows with postpartum fever, in primiparous cows, in cows with assisted parturition, and in cows that gave birth to twins. Our findings demonstrated that microbiota composition and TBL were associated with known periparturient risk factors of uterine diseases and reproductive failure, including parity, assisted parturition, and retained fetal membranes.

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³: Permanent Address: Department of Theriogenology, College of Veterinary Medicine, Al-Ameria, Baghdad, Iraq.

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Postpartum uterine flora following normal and abnormal puerperium in cows

Abstract

Theriogenology

Theriogenology

Theriogenology

Theriogenology

Theriogenology

The Normal Involution of Uteri in the Cow

Nord. Vet. Med.

Uterus of the Cow after Parturition. Bacterial Content

Am. J. Vet. Res.

Non-specific Uterine Infection and Bovine Fertility. I: Infection Patterns and Endometritis during the first Seven Weeks Postpartum

Theriogenology

Postpartum Evaluation of Bovine Reproductive Potential: Comparison of Findings from Genital Tract Examination per Rectum, Uterine Culture and Endometrial biopsy

J. Am. Vet. Med. Assoc.

Aspects of Bacteriology and Endocrinology of Cows with Pyometra and Retained Fetal Membrane

Am. J. Vet. Res.

Bovine Uterine Flora Post partum. A comparison of Swabbing and Biopsy

Vet. Rec.